Boletín de la SEA - Sociedad Entomológica Argentina | SEA

Transcription

N° 26 (1) 2015 Edición especial
ISSN 1666-4612
Boletín de la SEA
Sociedad Entomológica Argentina
Encontranos en:
http://seargentina.myspecies.info/
Facebook: Sociedad-Entomologica-Argentina
http://www.museo.fcnym.unlp.edu.ar/
De los Editores
Estimados lectores:
Este número especial del Boletín, está íntegramente dedicado al VIII International
Symposium on Phlebotomine Sandflies (ISOPS VIII). Esta publicación incluye 249
resúmenes de los trabajos presentados por los participantes del simposio, que fueron
sometidos a arbitraje oportunamente. Los resúmenes están agrupados en dos bloques,
uno correspondiente a las mesas redondas y ponencias orales, y otro a los posters. En
todos los casos se abarcan diferentes temáticas, incluyendo taxonomía, filogenia,
genética, ecología, control de vectores, entre otros. Las búsquedas se podrán realizar
a través de un índice temático y/o por autor. Por último se muestra una tabla con
información referida a la cantidad de autores por país y a la de presentaciones según
cada modalidad.
Esperamos que este número sea del interés de los lectores y agradecemos a los
organizadores del Simposio la elección de nuestro Boletín para publicar los
contenidos del mismo.
Hasta el próximo número
Las editoras
Dirección y edición
Nora Cabrera
Fabiana Gallardo
Edición y diseño
Julia Rouaux
Dirección
Boletín de la SEA. División
Entomología, Museo de La Plata.
Paseo del Bosque s/n.
La Plata CP(1900), Buenos Aires,
Argentina.
A los autores
A todos aquellos que deseen enviar
contribuciones para el Boletín por
favor dirigirse previamente a
Nora Cabrera:
ncabrera@fcnym.unlp.edu.ar
para recibir las pautas editoriales.
Dear Friends and Colleagues,
It is a great privilege for us to host and organize the VIII International Symposium on Phlebotomine Sandflies
in Puerto Iguazú, Argentina. This time the ISOPS features a Keynote Lecture by Dr. Eunice A. Bianchi Galati, 8
speakers at Round Tables and 73 Oral Presentations. In addition, 249 abstracts were presented, belonging to
813 authors from 35 countries.
As many of you know, because you have participated in this process, the first ISOPS took place in Rome in
1991 due to the impulse of Dr. Robert Killick-Kendrick and Dr. Michele Maroli, honoring the 300th
Anniversary of the first-known illustration of a Phlebotominae sand fly by Filippo Bonanni. After this former
event, ISOPS has alternated between the Old and the New World in Venezuela, France, Brazil, Tunisia, Peru,
and Turkey.
In 2014, this original idea reaches the eight Symposium in Argentina. After seven successful ISOPS we
recognize ourselves as a growing community, but without losing the feeling of a periodical gathering of
friends. Now we must preserve the heritage but also face the new challenges, and look forward with the main
objective to reduce the sand fly-borne diseases burden in a changing world.
New generations and methodologies are being incorporated; new voices were added to the multilateral global
complexity; new knowledge and so new questions come with each ISOPS.
As an ISOPS group, we can constitute a consolidated voice of sandfly researchers that could give evidencebased opinions, asked for or given to health policy makers. The consensual outcomes should include the voice
of leading experts but also the amazing experience of worldwide distributed groups.
As the ISOPS event, we can periodically produce a document that updates research-based evidence, gaps and
priorities, beyond objectives narrowed by individual, laboratory, country or 'technological fashion' interests.
As the ISOPS community, we can strength the collaborative network by searching for multi-partner projects
and cross-training empowerment internships, according to the priorities defined in the ISOPS document. We
can also establish common ground to define basic research protocols in order to make the results of different
groups more comparable, and so achieve faster conclusions and stronger recommendations.
We thought of many ways to homage the respected and beloved ISOPS friends that we have lost in the last
years: Dr. Bob Killick Kendric, Dr. Italo Sherlock, Dr. Richard Titus, and our friend Dr. Alexandre Peixoto.
Finally, we realized that the better way to make so is to keep the ISOPS going on and growing. Therefore, we
can further discuss the ideas presented here during ISOPS VIII.
We are very thankful to the Ministry of Health of Argentina, to all conference sponsors for their contributions,
to the members of the Honorary and Scientific Committees for their active participation, and to you for your
willing attendance.
We hope you have a fruitful ISOPS, and a pleasant stay in Puerto Iguazú, one of the seven Natural Wonders of
the World.
On Behalf of the Organizing Committee and the Argentinean Network for Research on Leishmaniasis
(REDILA)
Oscar Daniel Salomon
Maria Gabriela Quintana
Maria Soledad Santini
Honorary President Eunice A B Galati
HONORARY COMMITTEE
Añez Néstor
Killick Kendrick Mireille
Marolli Michele
Özbel Yusuf
Pérez Enrique
Rioux Jean-Antoine
Ready Paul D
Shaw Jeffrey
SCIENTIFIC COMMITTEE
Bates Paul
Brazil Reginaldo
Depaquit Jérôme
Elnaiem Dia-Eldin
Feliciciangeli Dora
Ferro Cristina
Galati Eunice
Gallego Montserrat
Lawyer Phillip G
Léger Nicole
Pimenta Paulo
Rangel Elizabeth
Valenzuela Jesús
Volf Petr
Warburg Alon
Yaghoobi-Ershadi Reza
ORGANIZING COMMITTEE
President
Oscar Daniel Salomón
Collaborators
Soraya Acardi
Mariana Manteca Acosta
Secretaries
Analia Araujo
Ma. Gabriela Quintana
Alicia Paola Benitez Ibalo
Ma. Soledad Santini
Pablo Berrozpe
Christina McCarthy
Treasury
Nicolás Silvero
José Direni Mancini
Magalí Giulinai
Javier Liotta
Accreditation and Diffusion
Beatriz Oscherov
Denise Fuenzalida
Julia Portnoy
Enrique Szelag
Cristina Remondegui
Mariela Martínez
Lucrecia Villarquide
Translations
Sergio Casertano
Graphic Design
Claudia Nose
María Eugenia Utgés
Website Design
Editorial and Publications
Juan Rosa
María Soledad Fernández
Matías Parra
Martín Abelardo
ISOPS VIII
Puerto Iguazú
Argentina
Content
&
Index
N° 26 (1) 2015
Boletín de la Sociedad Entomológica Argentina
CONTENT
ROUND TABLES AND ORAL SESSIONS
Phlebotomine Sand Flies (Diptera: Phlebotominae) in Brazil”
ID 1-O Albert Picado; Murari Lal Das; Mark Rowland; James
Austin; Elisa De Lazzari. “Evaluation of Different Nets Against
Phlebotomus Argentipes, The Vector of Visceral Leishmaniasis In
Nepal”
ID 2-O Rajesh B Garlapati, Sachidananda Samantaray, Mutum
Ingobi Singh, Trey Barresi, Dylan Burruss and Richard Poche.
“Multiple Fipronil Doses in Dairy Cattle for Vector Control in Bihar,
India: Residue Study and Efficacy Determination Against the Sand
Fly Phlebotomus Argentipes”
ID 3-O M Derbali, L Polyakova, A Boujaâma, D Burruss, S Cherni, W
Barhoumi, I Chelbi, R Poch, E Zhioua. “Laboratory and Field
Evaluation of Rodent Bait Treated with Fipronil for Feed Through and
Systemic Control of Phlebotomus papatasi”
6
ID 14-O V Carter, DP Bray, G Borges-Alves, E Dilger, O Courtenay,
R Brazil and JGC Hamilton. “Sex Pheromones as a Pre-Mating
Species Isolation Barrier in Brazil”
ID 15-O Chukwunonso Onyemaechi Nzelu, Abraham G Cáceres,
Ken Katakura, Yoshihisa Hashiguchi and Hirotomo Kato.
“Identification of Peruvian Sand Fly Species (Diptera: Psychodidae)
through DNA Barcodes”
ID 16-O Randrianambinintsoa Fano José; Léger Nicole; Boyer
Sébastien; Robert Vincent; Depaquit. Jérôme. “Origins and Affinities
of Malagasy Phlebotomine Sandflies”
ID 17-O Israel Cruz, Rosa Gálvez, Oihane Martín, Ivonne Pamela
Llanes-Acevedo, Carolina Arcones, Rocío Checa, Ana Montoya,
Carmen Chicharro, Silvia Miguelañez, Guadalupe Miró. “DNA
Barcoding of Mediterranean Phlebotomine Sandflies Based on the
Sequence Analysis of the Mitochondrial Cytochrome Oxidase I
Gene”
ID 4-O Mehmet Karakuş; Samiye Demir; Hüseyin Çetin; Suha Kenan
Arserim; Önder Ser; Seray Töz; I Cüneyt Balcioğlu; M Kirami Ölgen;
Bahtiyar Yilmaz; Yusuf Özbel.“Natural Infection and Insecticide
Susceptibility Status of Wild Caught Sand Flies in Rural Areas of
Antalya, Mediterranean Region of Turkey”
ID 18-O Jérôme Depaquit, Maria Dolores Bargues, Santiago MasComa. “A Molecular Phylogeny of the Phlebotominae”
ID 5-O David M Poché, William E Grant, Hsiao-Hsuan Wang. “Sand
Fly Control in Bihar, India: Modeling Alternative Control Strategies
Using Systemic Insecticides Orally Administered to Livestock”
ID 19-O Aysheshm Kassahun, Kristýna Hlavačková, Vít Dvořák,
Asrat Hailu, Alon Warburg, Jan Votýpka and Petr Volf. “Distribution,
Phylogeny and Taxonomy of Sergentomyia in Ethiopia”
ID 6-O Diwakar Singh Dinesh, Faizan Hassan, Vijay Kumar and
Pradeep Das. “Development of Resistance Against DDT to
Phlebotomus argentipes (Diptera: Psychodidae) and Search for
Alternative Insecticide”
ID 20-O Vit Dvořák, Jérôme Depaquit, Valentin Pflüger, Petr Halada,
Kristýna Hlavačková, Veronique Lehrter, Jorian Prudhomme, Sonja
Zapata, Petr Volf, Alexander Mathis. “Rapid Identification of
Phlebotomine Sand Flies by Maldi-Tof Mass Spectrometry”
ID 7-O Erika Santamaría, Olga Lucía Cabrera, Raul H Pardo.
”Toxic and Behavioural Effects in Experimental Huts of Long
Lasting Insecticide Treated Nets Against Lutzomyia Longiflocosa in
the Sub-Andean Rural Zone of Colombia”
ID 21-O Vit Dvorak, Ozge Erisoz Kasap, Jerome Depaquit, Bulent
Alten, Jan Votypka, Petr Volf. “Two New Species of the Subgenus
Transphlebotomus Revealed by Molecular Taxonomy”
ID 8-O Waleed Al Salem, Louise Kelly-Hope, Maha Abdeladhim,
Salah Balgonaeem, Mohammed Al Zahrani, Jesús Valenzuela, Álvaro
Acosta-Serrano. “A control strategy for Old World cutaneous
leishmaniasis outbreaks”
ID 22-O Ivonne Pamela Llanes-Acevedo, Carolina Arcones, Rosa
Gálvez, Oihane Martín, Rocío Checa, Ana Montoya, Carmen
Chicharro, Silvia Miguelañez, Susana Cruz, Guadalupe Miró, Israel
Cruz. “DNA Sequence Analysis Suggests that Cytb-Nd1 Pcr-Rflp
may not Be Applicable to Sandfly Species Identification in the Whole
Mediterranean Region”
ID 9-O Raúl H Pardo, Olga Cabrera, Catalina Marceló, Erika
Santamaría. “Long Lasting Insecticide Treated Nets Decrease
Sandfly Abundance and Blood Feeding Success Indoors in the SubAndean Region of Colombia: Results of a Hamlet Vector Control Trial
after One Year Follow Up”
ID 10-O Olga Lucía Cabrera, Erika Santamaría, Raúl H Pardo. “First
Experimental Hut to Study The Effect of Insecticide Control
Measures Against Sandflies (Diptera: Psychodidae)”
ID 11-O Manteca Acosta M; Molina J; Utgés M E; Mastrangelo AV;
Pérez AA; Santini M S; Salomón OD. “Efficacy Of Impregnated
Bednets And Species Composition In Experimental Henhouses”
ID 12-O Domenico Otranto; Filipe Dantas-Torres; Emanuele Brianti;
Fabrizio Solari Basano; Dorothee Stanneck; Katrin
Deuster.“Prevention of Canine Leishmaniosis in Naturally Exposed
Dogs Using a 10% Imidacloprid/ 4.5% Flumethrin Collar
(SERESTO®)”
ID 13-O Andrey J de Andrade, Paloma H F Shimabukuro, Eunice A B
Galati. “A New Proposal for the Taxonomic Identification of
ID 23-O Reale S, Torina S, Zaffora G, Cosenza M, Pitti R,
D'Agostino R, Intile S., Piazza M, and Vitale F. “Multilocus Real
Time PCR Identification of Phlebotomine Sand Flies Captured in
Southern Italy”
ID 24-O Nicole Léger and Jérôme Depaquit. “The Genus
Idiophlebotomus (Diptera; Psychodidae)”
ID 25-O Añez Néstor, Rojas Agustina, Yépez José Yancarlos. “Sand
Fly Species Transmitting Visceral Leishmaniasis at the Semiarid
Region of Venezuela”
ID 26-O Luis G. Estrada, Horacio Cadena, Edgar Ortega, Luz A
Acosta, Andrés Vélez-Mira, Rafael J Vivero, Eduar E Bejarano,
Sandra Uribe, Iván D Vélez. “Natural Breeding Sites of Lutzomyia
Evansi (Diptera: Psychodidae) in the Urban Zone of Ovejas, Sucre –
Colombia”
ID 27-O Perez JE, D Rado, D Quispe, A Quispe, F Toccas, M Chacon,
B Valencia, A Llanos, M Quispe, E Aguilar, R Pacheco & E Ogusuku.
“Lutzomyia Faunas (Diptera: Psychodidae) in the East Side if the
N° 26 (1) 2015
Andean Cordillera in Cuzco, Peru”
Özbel, P Volf, B Alten. “Phlebotomine Sand Flies Crossing Anatolia:
Do Borders Limit their Distribution?”
ID 28-O Karina Mondragon-Shem; Waleed Alsalem; Louise KellyHope; Maha Abdeladhim; Mohamed Alzahrani; Ziad Memish; Jesus
Valenzuela; Alvaro Acosta-Serrano.”Differential Exposure to Sand
Fly Bites in Saudi Arabia”
ID 29-O Paulo Silva de Almeida, Alan Sciamarelli, Josué Raizer,
Jaqueline Aparecida Menegatti, Sandra Cristina Negreli Moreira
Hermes, Maria do Socorro Laurentino de Carvalho, Andrey José de
Andrade, Rodrigo Gurgel-Gonçalves. “Predicting the Geographical
Distribution of Leishmaniasis Vector Species in Central West Brazil”
ID 30-O Raquel Gonçalves, Daniela Cristina Soares, Ricardo
Guimarães, Walter Souza Santos, Gilberto Cesar de Sousa, Anadeiva
Portela Chagas, Lourdes Maria Garcez. “Ecology of Sandflies in Risk
Zones for Tegumentary Leishmaniasis Transmission in Santarém
Municipality, Western Pará State, Brazil”
ID 31-O Dia-Eldin A Elnaiem. “Phlebotomus Orientalis and the Fall
and Rise in Visceral Leishmaniasis in East Africa”
ID 32-O Bilel Chalghaf, Amine Toumi, Sadok Chlif, Jihene Bettaieb,
Mariem Harrabi, Goze Bertin Bénié, Afif Ben Salah. “Distribution of
Leishmaniasis Vectors in the Mediterranean Basin in the Light of
Climate Change”
ID 33-O Aviad Moncaz, Oscar Kirstein, Araya Gebresellassie,
Wossenseged Lemma, Solomon Yared, Teshome Gebre-Michael,
Asrat Hailu, Moshe Shenker and Alon Warburg. “Characterization of
Breeding Sites of Phlebotomine Sand Flies in the Judean Desert and
North-West Ethiopia”.
ID 34-O Oscar David Kirstein, Araya Gebreselassie, Aviad Moncaz,
Asrat Hailu, Teshome Gebre-Michael and Alon Warburg. “Fine Scale
Analyses of Factors Contributing to the Transmission of Visceral
Leishmaniasis in North Ethiopian Villages: An Eco-Epidemiological
Approach”
ID 35-O Wossenseged Lemma, Habte Tekie, Meshesha Balkew,
Teshome Gebre-Michael3, Alon Warburg, Asrat Hailu. “SeroPrevalence L. donovani in Labour Migrants Population dnd
Entomological Risk Factors in Extra-Domestic Habitats of MetemaHumera Lowlands - Kala-Azar Endemic Areas in the Northwest
Ethiopia”
ID 36-O Araya Gebresilassie1, Ibrahim Abbasi2, Oscar David
Kirstein2, Aviad Moncaz2, Habte Tekie1, Meshesha Balkew3, Alon
Warburg2, Asrat Hailu4, and Teshome Gebre-Michael3 “Host
Preference Patterns of Phlebotomus (Larroussius) orientalis Parrot,
1936 (Diptera: Psychodidae) in the Visceral Leishmaniasis Endemic
Area of Tahtay Adiyabo District, Northern Ethiopia”
ID 37-O Esayas Aklilu, Habte Tekie, Meshesha Balkew, Alon
Warburg, Asrat Hailu, Teshome Gebre-Michael. “Bionomics of
Phlebotomine Sandflies in a Highland Kala-Azar Focus in LiboKemkem District, Northwestern Ethiopia”
7
ID 40-O M R Yaghoobi-Ershadi, M Darvishi, F Shahbazi, AA
Akhavan, R Jafari, M Khajeian, H Soleimani, H Darabi, M H
Arandian. “Epidemiological Study on Sand Flies in an Endemic
Focus of Cutaneous Leishmaniasis, Bushehr City in Southwestern
Iran”
ID 41-O Jeffrey Jon Shaw. “Eco-Epidemiology of Leishmaniases:
Vectors and New Scenarios of Transmission.”
ID 42-O Alon Warburg. “Emerging Cutaneous Leishmaniasis in
Israel: A Review of The Last Two Decades”
ID 43-O Ns Singh; Doris Phillips-Singh; Ipe M Ipe; Shubham
Pandey; Dinesh Lal. “Studies on the Sand Fly Fauna (Diptera:
Psychodidae) in High-Transmission Areas of Visceral Leishmaniasis
in North India, Special Emphasis on Uttaranchal Region”
ID 44-O W Fares, RN Charrel, K Dachraoui, L Bichaud, W
Barhoumi, M Derbali, S Cherni, I Chelbi, X de Lamballerie, E
Zhioua. “Infection of Wild Sandflies Collected from Different BioGeographical Areas of Tunisia with Phleboviruses”
ID 45-O Bongiorno G, Lisi O, Severini F, Vaccalluzzo V, Khoury C,
Di Muccio T, Gradoni L, Maroli M, D'urso V, Gramiccia M,
“Investigations On Sand Fly Bionomics and Leishmania Natural
Infections in Eastern Sicily, Italy, with Particular Reference to
Phlebotomus Sergenti”
ID 46-O Alveiro Pérez-Doria, Margaret Paternina, Lily Martinez,
Luis Paternina, Daniel Verbel-Vergara, Suljey Cochero, Eduar E.
Bejarano, “Natural Infection of Lutzomyia Cayennensis Cayennensis
(Diptera: Psychodidae) with Leishmania (Viannia) spp., Potential
Vector in the Enzootic Cycle of Leishmaniasis in Urban And Rural
Habitats of The Colombian Caribbean”
ID 47-O Paul D Ready “Priorities for Progressing from Assessments
of Vector Competence to the Transmission Modelling and Control of
Leishmaniasis”
ID 48-O Paulo F P Pimenta, Vanessa C Freitas, Nágila F C
Secundino. “Leishmania Chagasi in its Natural Vector Lutzomyia
longipalpis.”
ID 49-O Jovana Sadlova, Veronika Hrobarikova, Jan Votypka, Alon
Warburg and Petr Volf. “Xenodiagnosis of Leishmania donovani
Infections in Asymptomatic Mice Using Phlebotomus orientalis Sand
Flies
ID 50-O V Seblova-Hrobarikova, J Sadlova, S Carpenter, P. Bates
and P Volf. “Development of Leishmania Parasites in Biting Midges,
Culicoides nubeculosus and Culicoides sonorensis (Diptera:
Ceratopogonidae) and Implications for Screening Vector
Competence”
ID 38-O Solomon Yared, Meshesha Balkew, Alon Warburg, Asrat
Hailu and Teshome Gebre-Michael. “Species Composition,
Abundance and Seasonal Dynamics of Phlebotomus Species in a
Visceral Leishmaniasis Endemic Area of Northwest Ethiopia”
ID 51-O Luis Roberto Romero, Jorge Luis Rodríguez, Alveiro PérezDoria, José G. Vergara, Ana Montesino, Lily Paola Martínez, Luis
Cortés, Suljey Cochero & Eduar Elías Bejarano. “The Sand Fly
Lutzomyia evansi (Diptera: Psychodidae), as a Permissive Vector of
Leishmania Infantum and Le. braziliensis In Bolívar, Colombia”
ID 39-O O Kasap Erisoz, A Belen, C Alkan, F Gunay, V Dvorak, K
Ergunay, S Aydın, J Votypka, A-L Banuls, R Charrel, A Özkul, Y
ID 52-O Ana Clara Araújo Machado Pires, Paulo Filemon Paolucci
Pimenta and Nágila Francinete Costa Secundino. “Experimental
N° 26 (1) 2015
Susceptibility of Lutzomyia (L.) Longipalpis to Different Species of
Leishmania”
ID 65-O Anayansi Valderrama Cumbrera, Mara Garcia Tavares,
Ingrid Murgas, José Andrade Dilermando. “Genetic Assessment of
Sandfly Populations in Panama”
ID 53-O Shaden Kamhawi, Fabiano Oliveira and Jesus Valenzuela
“The Prospect for a Sand Fly Saliva-Based Vaccine Against
Leishmaniasis in Humans”
ID 54-O Pitaluga AN, Tinoco B, Di Blasi T, Silva AM, Tempone A,
Traub-Csekö YM. “Characterization of Lutzomyia Longipalpis
Immune Responses with Emphasis in Leishmania Infection”
ID 55-O Matthew E Rogers, Emilie Giraud, Rod J Dillon, Paul A
Bates, Tamsyn Derrick, Oihane Martin, Ingrid Műller. “Promastigote
Secretory Gel from Infected Sand Flies Subverts the Wound Healing
Response in an Insulin Growth Factor1-Dependent Manner”
ID 56-O Ranadhir Dey, Hamide Aslan, Amritanshu Joshi, Claudio
Meneses, Pradeep K. Dagur, John Philip McCoy, Robert Duncan,
Jesus G. Valenzuela, Hira L. Nakhasi and Shaden Kamhawi, “VectorTransmission of Leishmania donovani Triggers a Distinct Immune
Response that Favors Parasite Survival and Dissemination”
ID 57-O Wafa Rebai-Kammoun, Ikbel Naouar, Darragh Duffy,
Valentina Libri, Milena Hasan, Amine Toumi, Afif B. Salah, Matthew
Albert, Hechmi Louzir, Amel Meddeb-Garnaoui. “Cytokine Levels
in Response to Salivary Gland Extracts and to Leishmania major
Parasites in Individuals Living in Endemic Areas for Zoonotic
Cutaneous Leishmaniasis in Tunisia”
ID 58-O Ikbel Naouar, Amel Meddeb-Garnaoui, Amine Toumi, Nabil
Belhadj Hamida, Afif B Salah, Hechmi Louzir. Presenting author at
the event: Wafa Rebai Kammoun. “Prospective Study of a Cohort of
Individuals at Risk of Zoonotic Cutaneous Leishmaniasis in Tunisia
to Evaluate Clinical and Immunological Parameters and Identify
Correlates of Protection.”
ID 59-O Tatiana Di-Blasi; Antonio Jorge Tempone; Erich Loza
Telleria; Fabricio K Marchini; José Marcelo Ramalho-Ortigão; Yara
Maria Traub-Csekö. “The Possible Role of a Flagellar Protein from
Leishmania sp. (FLAG) in the Parasite - Sand Fly Interaction”
ID 60-O Votypka J, Pruzinova K, Hlavacova J, Volf P. “The Effect of
Temperature and Avian Blood on Leishmania Development in Sand
Flies”
8
ID 66-O Noteila M Khalid, Marium A Aboud, Fathi M Alrabba, DiaEldin A Elnaiem and Frederic Tripet. ”Evidence for Genetic
Differentiation at the Microgeographic Scale in Phlebotomus
papatasi Populations from Sudan”
ID 67-O Hector M Diaz-Albiter, Caroline d Moraes, Maiara d Faria,
Mauricio R Sant'Anna, Roderick J Dillon and Fernando A. Genta.
“Gene Expression Profiling and Silencing of Key Digestive Enzymes
in Lutzomyia Longipalpis”
ID 68-O Remoli ME, Bongiorno G, Fortuna C, Marchi A, Bianchi R2,
Khoury C, Ciufolini MG, Gramiccia M. ” Evaluation of Collection
Methods for Phlebotomus-Borne Viruses Detection: Isolation and
Viral RNA Integrity Performance”
ID 69-O Andrea Martins da Silva, Antonio Jorge Tempone, André N
Pitaluga, Fabricio K Marchini, Barbara Burleigh, Yara M TraubCseko. “Conditioned Media and Exosomes from Lutzomyia
longipalpis LL5 Cells dsRNA-Transfected are able to Induce NonSpecific Antiviral Response in LL5 Naive Cells”
ID 70-O Meneses C, Morales E, Rhodes C, Oliveira F, Valenzuela
JG and Kamhawi S. “CD-Rom Sand Fly Fellas: The First Complete
Video-Based Sand Fly Rearing Guide in High Definition”
ID 71-O Vicente Estevam Machado; Thais Marchi Goulart; Arlene
Gonçalves Corrêa; Mara Cristina Pinto. “Comparison of Volatile
Compounds Emitted by Animals or Present in Plants on
Attractiveness of Nyssomyia neivai (Diptera:Psychodidae)
ID 72-O Ifhem Chlebi, Elyes Zhioua, and Gordon Hamilton.
“Responses of the Sand Fly Phlebotomus papatasi Scopoli (Diptera:
Psychodidae) to an Oviposition Pheromone Associated with
Conspecific Eggs”
ID 73-O Vivero J Rafael; Cadavid Gloria; Moreno H Claudia
Ximena; Posada-López Laura; Sandra Uribe S. “Study on the
Bacterial Midgut Microbiota Associated to Immature Stage of Genus
Lutzomyia (Diptera: Psychodidae) Isolated from Natural Breeding
Sites”
ID 61-O Mary Ann McDowell and the Sand Fly Genome Consortium.
“Two Sand Fly Genomes: Phlebotomus papatasi And Lutzomyia
longipalpis”
POSTERS
ID 62-O Gloria I Giraldo-Calderón, Scott J. Emrich, Daniel Lawson,
Frank H Collins and the members of the VectorBase consortium. “The
Database Vectorbase and the Genomes of Lutzomyia longipalpis and
Phlebotomus papatasi”
ID 1-P Sirlei Franck Thies, Ana Lucia Maria Ribeiro, Jorge
Senatore Rodrigues Vargas, Edelberto Santos Dias and Amilcar
Sabino Damazo “Sandfly fauna (Diptera: Psychodidae) of urban area
in the city of sinop, Mato grosso, Brazil”
ID 63-O Samuel Tadros, Mariha Wadsworth, Mallory Hawksworth,
RJ Nowling, Mary Ann
McDowell, Gloria I Giraldo-Calderón. “Manual Annotation of G
Protein–Coupled Receptors (GPCRs) in the Genomes of Lutzomyia
longipalpis and Phlebotomus papatasi”
ID 2-P Sirlei Franck Thies, Marinês Browers “Sandfly fauna
(Diptera: Psychodidae) from regional of health in Sinop, Mato
Grosso, Brasil, year 2013.”
ID 64-O Gabriela Vela, Gabriel Trueba, Jérôme Depaquit, Sonia
Zapata. “Genetic Variation of Nyssomyia trapidoi (Diptera:
Psychodidae) in Three Endemic Zones of Leishmaniasis in the
Ecuador”
ID 3-P Lorena G. Caligiuri, Enrique A. Sandoval, María Soledad
Santini, Soraya Acardi, Oscar D. Salomón, Lilian Tartaglino and
Christina B. McCarthy “Testing the versatility of a Psychodiella sp.
diagnostic assay in field surveys”
ID 4-P Inés Martín-Martín, Ricardo Molina and Maribel Jiménez
“Kinetics of anti-Phlebotomus perniciosus saliva antibodies in mice
and rabbit será”
N° 26 (1) 2015
ID 5-P Laura W. Juan, Ignacio T. Gould, Oscar D. Salomón, Juan
Molina, Raul A. Alzogaray and Eduardo N. Zerba “Field evaluation
of an insecticide formulated with active ingredients acting on larval
and adult stages of Lutzomyia longipalpis”
Visceral Leishmaniasis in northeast of Brazil: Fortaleza, state of
Ceará, Brazil”
ID 6-P Douglas De Almeida Rocha, Thaís Oliveira Coelho,
Rodrigo Gurgel Gonçalves, Andrey José de Andrade and Marcos
Takashi Obara “Characterization of susceptibility of populations of
Lutzomyia longipalpis (Diptera: Psychodidae) to
alphacipermetrin”
ID 7-P Tobin Rowland, Silas Davidson, Kevin Kobylinski and
Edgar Rowton “Effects of Leishmania major Infection on Sand Fly
Feeding Responses to DEET and or permethrin treated uniform
material”
ID 8-P Horacio Cadena, Edgar Ortega, Luz A. Acosta, Andrés
Vélez-Mira, Rafael J. Vivero,Eduar E. Bejarano, Sandra Uribe,
Hernando Gómez, Luis G. Estrada and Iván D. Vélez “Soil analysis
and vegetation associated with the breeding sites of Lutzomyia evansi
in the municipality of Ovejas-Sucre a focus of leishmaniasis in
Colombia”
9
ID 17-P Fernandes Brilhante A., Galati E. A. B, de Oliveira Cardoso
C., Progênio dos Santos N., Ferreira de Souza J. and Vidal J.
“Phlebotominae captured in area of transmission of American
cutaneous leishmaniasis (ACL) in Xapuri municipality, Acre state,
Western Brazilian Amazonian. Preliminary results”
ID 18-P dos Santos Afonso M. M., de Miranda Chaves S. A., de
Avelar Figueiredo Mafra Magalhães M., Garrido R., Graser C., de
Carvalho B. M., Costa S. M. and Rangel E. F. “The geographic
expansion of American Visceral Leishmaniasis associated with
deforestation in Tocantins State, Brazil”
ID 19-P dos Santos Afonso M. M., de Miranda Chaves S. A., de Pita
Pereira D., da Costa Rego T. A., de Santana A. L. F., dos Santos Silva
J., de Carvalho B. M., da Costa S. M., Giordano C. and Rangel E.
F.“Lutzomyia (Lutzomyia) longipalpis feeding habits and natural
infection by Leishmania (Leishmania) infantum chagasi in some
Brazilian endemic areas of American visceral leishmaniasis”
ID 9-P David Poché, Alex Gendernalik, Larisa Polyakova, Richard
Poché, Madina Kozhahmetova, Batirbek Aimakhanov, Ziyat
Abdeliyev and Bakhit Atshabar “The use of systemic insecticides to
control Phlebotomus spp. sand flies in southern Kazakhstan”
ID 20-P Stramandinoli Moreno E., Posteraro Freire M., Cordeiro
Nascimento L. O., Zanini V., Ribeiro Alves V. , Fernandes
Shimabukuro P. H. “Sand flies (Diptera, Psychodidae,
Phlebotominae) collected in the Wajãpi Indigenous Reserve, Eastern
Amazon, Brazil”
ID 10-P José Carlos C B Ribeiro, Antônio Menezes Filho, Daniela M
Parente, Teresinha C Farias, Danielle A Zacarias, Jalthon C da Silva,
Dorcas L. Costa, Ivete L Mendonça, Carlos H N Cost “Transmission
of Leishmania infantum to Lutzomyia longipalpis by humans with
and without HIV infection, symptomatic and asymptomatic”
ID 21-P Fernández M. S., Santini M. S., Cavia R. and Salomón O. D.
“Lutzomyia longipalpis (Leishmania infantum vector) and
Nyssomyia whitmani (Leishmania braziliensis vector) in the city of
Puerto Iguazú, northeastern Argentina: seasonal abundance and
relationship with domestic animals”
ID 11-P Quintana MG., Santini MS, Britez N, Rojas de Arias A,
Thomaz-Soccol V, Bisetto A, Bazzani R, Yadon Z, Salomón OD and
the IDRC Project #107577 team “Addressing the emergence and
spread of leishmaniasis in the borders of Argentina, Brazil and
Paraguay”
ID 22-P Machado C. A., Ribeiro Alves V. and Fernandes
Shimabukuro P. H. “Geographical distribution of Nyssomyia
Barretto, 1962 in Brazil”
ID 23-P Rosa J. R., Parras M. A., Szelag E. A and Salomón O. D.
“Vertical and horizontal distribution of sand flies in an area of
Tegumentary Leishmaniasis in Chaco, Argentina”
ID 12-P Fitipaldi Veloso Guimarães V. C., Lemos Costa P., da Silva F.
J., Lopes de Melo F., Dantas-Torres F., Gomes Rodrigues E. E. and
Pinto Brandão-Filho S. “Molecular detection of Leishmania in
phlebotomine sand flies in a cutaneous and visceral leishmaniasis
endemic area in northeastern Brazil”
ID 24-P Parras M. A., Rosa J. R., Szelag E. A. and Salomón O. D.
“Natural breeding sites of Phlebotominae (Diptera: Psychodidae) in a
flood zone of Chaco, Argentina”
ID 13-P Menezes, J.A., Sousa A. M., Morais M. H. G., Andrade-Filho
J. D., Lima F. P., Fux B. and Margonari C. “Study of sand fly fauna in
recent area transmission of leishmaniasis in the state of Minas Gerais,
Brazil”
ID 25-P Carvalho G.M.L., Quaresma P. F., Gontijo C. M. F., Rego F.
D., Rugani J. M. N., Dias T. A. and Andrade Filho J. D. “Analysis of
sand fly (Diptera: Psychodidae) bloodmeals using sequencing
technique: test of sand fly experimentally fed”
ID 14-P de Andrade A. J. and Gurgel-Gonçalves R. “New record and
update on the geographical distribution of Pintomyia monticola
(Costa Lima, 1932) (Diptera: Psychodidae: Phlebotominae) in South
America”
ID 15-P Rocha Dos Santos C. F., Lima M. C., Da Rocha Neto G. B.
and Moreira Macena A. L. “Entomological monitoring of the
programme of visceral leishmaniasis in Maceió, Alagoas State,
Brazil: indicators and environmental risk factors”
ID 26-P Acel A. M., Ribeiro A. L. M., Miyazaki R. D. and Senatore
Vargas Rodrigues J. “Sandfly survey in ecotourism region in the
district of Bom Jardim from the City of Nobres, Mato Grosso –
Brazil”
ID 27-P Ribeiro A. L. M., Senatore Vargas Rodrigues J., Parreiras de
Almeida T., Marques de Souza E., Pereira Barreto Bettiol F. K., Acel
A. M, Leal dos Santos F. A., Franck Thies S. and Dunjko Miyazaki R.
“Leishmaniasis sandfly diversity in a rural area of the Cuiabá, Mato
Grosso, Brazil”
ID 16-P Silva R. A., Bevilaqua C. M. L., Rangel E. F., Santos F. K.M.,
Sousa L. C., Donato L. E. and Pessoa G. C. D. “Aspects of the ecology
of Lutzomyia longipalpis and Lutzomyia migonei (Diptera:
Psychodidae: Phlebotominae) in area of intense transmission of
ID 28-P Dunjko Miyazaki R., Ribeiro A. L. M., Senatore Vargas
Rodrigues J., Parreiras de Almeida T., Marques de Souza E., Pereira
Barreto Bettiol F. K. and Acel A. M. “Incidence of sandflies at the
zoological of Federal University of Mato Grosso, Cuiabá/MT-Brazil”
N° 26 (1) 2015
ID 29-P Lemma W., Tekie H., Balkew M., Gebre-Michael, Warburg
A. and Hailu A. “Population dynamics and habitat preferences of
Phlebotomus orientalis in extra-domestic habitats of Metema Humera lowlands - kala-azar endemic areas in northwest Ethiopia”
ID 42-P da Costa S. M., Cordeiro J. L. P, Afonso M.S.11 and Rangel
E. F. “Habitat Suitability for Lutzomyia (Nyssomyia) whitmani and
American Cutaneous Leishmaniasis (ACL) in Brazil”
ID 30-P Lemos Costa P., Dantas-Torres F., José da Silva F., Fitipaldi
Veloso Guimarães V. C., Gaudêncio K. and Pinto Brandão Filho S.
“Ecology of Lutzomyia longipalpis in an area of visceral
leishmaniasis transmission in north-eastern Brazil”
ID 31-P de Oliveira Miranda D. E., Gaudêncio da Silva K., da Gloria
Faustino M. A., Câmara Alves L., Pinto Brandão-Filho S., DantasTorres F. and de Carvalho G. A. “Diversity of phlebotomine sand flies
in an american cutaneous leishamaniasis endemic area in northeastern brazil”
ID 32-P Gaudêncio da Silva K., Silva de Morais R. C., de Oliveira
Miranda D. E., de Arruda Moura A. C., Pinto Brandão-Filho S., de
Paiva Cavalcanti M. and Dantas-Torres F. “A real-time PCR for
characterization of multiple blood sources in field-collected
phlebotomine sand flies”
ID 33-P Maia C., Cristóvão J., Freitas F. B., Afonso M.O. and
Campino L. “A three consecutive years phlebotomine sand fly survey
in a zoonotic leishmaniasis endemic focus, the Algarve Region,
Portugal”
ID 34-PLemma W, Tekie H, Abassi I, Meshesha Balkew M., GebreMichael T., Warburg A. and Hailu A. “Nocturnal Activities and Host
Preferences of P. orientalis in Extra-Domestic Habitats of Kala-Azar
Endemic Metema–Humera Lowlands, North West Ethiopia”
ID 35-P Rigg C., Valderrama A, Calzada J.E., Chavez L. F. and
Saldaña A. “Prevalence of Leishmania spp. infection and host - blood
feeding of sandflies transmitters of cutaneous leishmaniasis in an
endemic locality in the district of capira, Panama”
ID 36-P Arserim S.K., Karakus M. and Ozbel Y. “A study on nocturnal
activity of sand flies in a leishmaniasis endemic village located in
Aydin province of Turkey”
ID 37-P Arserim S. K., Mermer A., Balcioglu I.C., Ermis V. O. and
Ozbel Y. “Seasonal Activities and Ecological Features of Sand Flies
(Diptera: Psychodidae: Phlebotominae) in Aydın Mountains and
Surroundings Located in Western Turkey, Endemic Area for
Cutaneous Leishmaniasis”
ID 38-P Araujo A. V., Benítez Ibalo A. P., Rosa J. R., Oscherov E. B.
and Salomón O. D. “Bionomy of Phlebotomine sandflies
(Phlebotominae) in an outbreak area of visceral leishmaniasis in
Corrientes, Argentina”
10
ID 43-P Gould, I. T., Fernández M. S., Pérez A A. and Salomón O. D.
“A field study to evaluate the performance of two modified CDC light
traps for collecting sandflies compared with the traditional CDC trap
light”
ID 44-P Gómez Bravo A., Gould I. T., Abril M., Salomón O. D.
“Population composition and dynamics of Lutzomyia longipalpis and
other Phlebotominae species in Clorinda, Northern Argentina”
ID 45-P Acardi S. A, Steinhorst I. I., Santini M. S., Sandoval A. E.,
Salomon O. D. “Description of population dynamics and hourly
activity of Lutzomyia longipalpis in the city of Posadas, Argentina”
ID 46-P Mondragon-Shem K., Bell M., Suar M., Routray P., Posada
L. and Cameron M “Comparison of CDC light and gravid traps for the
collection of sandfly fauna in Bhubaneswar, Orissa (India)”
ID 47-P Ilango Kandan “Taxonomy and diagnostic characters of the
Phlebotomus argentipes species complex”
ID 48-P Rodrigo E. Godoy, Eunice A. B. Galati, Nataly A. de Souza,
Thiago V. dos Santos, Lindemberg Caranha de Sousa, Adriana
Zwetsch and Elizabeth F. Rangel “Discriminant analysis as a possible
tool to identification Sand flies (Diptera: Psychodidae) species
through morphometric data”
ID 49-P Jomaa Chemkhi, Souheila Guerbouj, Ikram Guizani and Afif
Ben Salah “First report of abnormal numbers of spermathecae in a
female Phlebotomus (Larroussius) longicuspis (Diptera:
Psychodidae) in Tunisia”
ID 50-P D. de Almeida Rocha, J. Barbosa C. Ferreira, R. GurgelGonçalves and Andrey J. de Andrade “First record of Psathyromyia
pradobarrientosi (Le Pont, Matias, Martinez & Dujardin, 2004)
(Diptera: Psychodidae: Phlebotominae) in Brazil”
ID 51-P Priscila B. Sábio, Andrey J. de Andrade and Eunice A. B.
Galati “Records of females Psathyromyia cuzquena Martins, Llanos
& Silva, 1975 (Diptera, Psychodidae, Phlebotominae) in northern
Brazil”
Galati “Assessment of the taxonomic status of Phlebotomus pifanoi
Ortiz, 1972 (Diptera, Psychodidae, Phlebotomina
ID 53-P Nil Rahola " Uncharted Phlebotomine sand flies in Gabon”
ID 39-P Kirstein O. D., Faiman R., Gebreselassie A., Hailu A, GebreMichael T. and Warburg A. “Attraction of Ethiopian phlebotomine
sand flies (diptera: psychodidae) to light and sugar-yeast mixtures
(Co2)”
ID 54-P Patrick F. Botelho, Fábio M. Medeiros da Silva, Rodrigo E.
Godoy and Thiago V. dos Santos “Morphometric study on
populations of Psychodopygus complexa and Ps. Wellcomei (Diptera:
Psychodidae), vectors of cutaneous leishmaniasis in Brazil”
ID 40-P Moreira de Carvalho B., Moncassim Vale M., Lorini M. L.,
dos Santos Afonso M. M. and Rangel E. F. “Ecological niche
modeling predicts southward expansion of Lutzomyia flaviscutellata,
vector of diffuse cutaneous leishmaniasis in South America, under
climate change”
ID 55-P Veracilda R. Alves, Claudio Ruy V. da Fonseca and Paloma
H. F. Shimabukuro “Taxonomic review of Psychodopygus
nangabeira, 1941 (Diptera, Psychodidae, Phlebotominae
ID 41-P Kirstein o. D., Abassi I., Gebreselassie A., Gebre-Michael T.,
Hailu A. and Warburg A. “Identification of Plants fed upon by
phlebotomine sand flies in Ethiopia”
Galati “Taxonomic revision of the subgenus Psathyromyia Barretto,
1962 (Diptera: Psychodidae: Phlebotominae)”
ID 57-P Maia C., Parreira R., Cristóvão J., Afonso M.O. and Campino
N° 26 (1) 2015
L. “DNA Barcoding for the Identification of Sand Fly Species
(Diptera, Psychodidae, Phlebotominae) in Portugal”
longipalpis in henhouses in an American visceral leishmaniasis
endemic area of São Paulo State”
ID 58-P Casertano S.A., Andrade Filho J.D. and Salomon O.D. “New
records of Phlebotominae (Diptera: Psychodidae) from the Atlantic
Forest region of Argentina”
ID 72-P Ferreira Souza C., Quaresma P. F., Dilermando Andrade
Filho J., Peçanha Brazil R. and Dias Bevilacqua P. “Micropygomyia
quinquefer (Dyar, 1929) (Diptera: Psychodidae: Phlebotominae): a
puctative vector of Leishmania spp. ?”
ID 59-P Pérez-Doria A., Romero-Ricardo L., Lastre N. and Bejarano
Eduar E. “A method for extraction of genetic material which permits
the integration of classical taxonomy with identification based on the
DNA barcode”
ID 60-P Cristian F. Souza, Reginaldo P. Brazil, José D. Andrade
Filho, Paula D. Bevilacqua, Patrícia F. Quaresma and Mirelle B.
Souza “Contribution to the knowledge of the fauna of sand flies
(Diptera: Psychodidae: Phlebotominae) of Rio Doce State Park,
Minas Gerais, Brazil”
ID 61-P Amini Esfanjani MH, Barazesh A, Mazloumi Gavagni A and
Saeidi Z “Sand fly fauna in Kaleybar districts, northwestern
Iran”
ID 62-P Mierez M. L., Rea M. J. F., Borda C. E. and Mosqueda L. A.
“Malformations of the genitalia in male Micropygomyia quinquefer
(Dyar, 1929) (Diptera: Psychodidae) in Corrientes, Argentina”
ID 63-PPosada-López L., Vivero R., Uribe S. I., Acosta L. A., VélezMira A., Cadena H., Estrada L. G. , Ortega E., Bejarano E. and Vélez
I. D. “The use of DNA Barcoding for taxonomic identification of the
immature stages of phlebotomine sandflies in ovejas, sucre
(Colombia)”
ID 64-P Fernandez R., Zorrilla V., Espada L., Vásquez G. and Stell F.
“Sand flies (Diptera: Psychodidae) from Peru: report of new
records”
11
ID 73-P Kratochvílová T., Košťálová T., Kindlová M., Sumová P.,
Šíma M., Rohoušová I., Gramiccia M., Gradoni L., Hailu A., Baneth
G., Warburg A.
and Volf P.“Canine antibody response to
Phlebotomus perniciosus and Phlebotomus orientalis bites”
ID 74-P de Oliveira Lara-Silva F., Monteiro Michalsky E., Latorre
Fortes-Dias C., de Oliveira Pires Fiuza V., Marques Pessanha J.E., da
Silva S. R., Moreira de Avelar D., Alves Silva M., Vianna Mariano da
Rocha Lima A.C., da Costa A. J. A., Machado-Coelho G. L. L. and
Edelberto Santos Dias E. “Epidemiological aspects of vector,
parasite, and domestic reservoir in area with no reported human cases
of visceral leishmaniasis in Brazil”
ID 75-P Monteiro Michalsky E., Valadão Lopes J., Oliveira Lara E
Silva F., Alves SilvaM., Vianna Mariano Da Rocha Lima A. C.,
França-Silva J. C., Rodrigues Da Paixão F., Antunes Da Costa A. J.
and Santos Dias E. “Sandflies fauna survey and DNA detection of
Leishmania infantum in Lutzomyia longipalpis captured in the city of
Sabara, Minas Gerais, Brazil”
ID 76-Pde Nazaré Martins da Silva A., Ishikawa E. A. Y., Fadul M.,
Vasconcelos dos Santos T., Costa Vasconcelos dos Santos Y.,
Carvalho de Araújo P. T. and Queiroz Santos N. “Phlebotomine sand
flies (Diptera: Psychodidae) in three municipalities of the state of
Pará, Brazil, area of very high transmission of cutaneous
leishmaniasis”
ID 65-P Fuenzalida AD, Direni Mancini JM, Salomon OD, and
Quintana MG “First record of the genus Pintomyia Costa Lima, 1932
(Diptera Phlebotominae) in the northwest of Argentina”
ID 77-P Moreira Macena A L., Cavalcante De Matos Rodarte C.,
Tavares Da Silva M. E., Rocha Dos Santos C. F., Carneiro Lima M.
and Braga Da Rocha Neto G. “Preliminary study of sandflies in the
city of Maceió-Alagoas-Brazil”
ID 66-P Reis, A. S., Saraiva, L., Gontijo, C.M.F., Andrade Filho, J. D.
" Morphometric study of Lutzomyia longipalpis populations
(Diptera: Psychodidae) collected in urban and environmental
protected areas in Minas Gerais, Brazil”
ID 78-P de Morais Oliveira A. F., Lima Batista H., Azevedo Leite K.
C. and Nunes Lacerda H. M. “Education: demographic and social
impacts on the rate of incidence of Visceral Leishmaniasis in the State
of Tocantins, Brazil (2010)”
ID 67-P Almazán M. C., Gil J. F., Chanampa M. M., Barroso P.,
Nasser J, Copa G. N., Oreste F., Castillo G. “Sand flies captures and
identification of Leishmania subgenus in Giemsa-Stained slides of
patients from five localities of Salta, Argentina”
ID 79-P Silva Lana R., Monteiro Michalsky E., França Silva J. C., de
Oliveira Lara e Silva F., Latorre Fortes Dias C., Martins J. and Dias E.
S. “Eco-epidemiology of leishmaniasis in the region of Serra do Cipo,
a major touristic centreof Minas Gerais”
ID 68-P Szelag E. A., Rosa J. R., Parras M. A. and Salomón O. D
“Phlebotominae Bionomics in the Humid Chaco Bio-region. Chaco,
Argentina”
ID 80-P de Moraes S. C., Lane Melo A. and Rangel E. F. “Survey of
sandflies (Diptera: Psychodidade: Phlebotominae) in indoor
environments, peridomestic and extra-domestic transmission area of
American cutaneous leishmaniasis (ACL) and American visceral
leishmaniasis (AVL) in Mato Grosso, Brazil”
ID 69-P Szelag E. A., Parras M. A., Rosa J. R. and Salomón O. D
“Dispersion of Lutzomyia longipalpis in the Chaco province,
Argentina”
ID 70-P Galvis Ovallos F., da Paixão Sevá A., Galucci E., Martins
Soares R., Ferreira F. and Galati E. A. B. “Monthly distribution of
Lutzomyia longipalpis in an endemic area of visceral leishmaniasis in
São Paulo state, Brazil”
ID 71-P Galvis Ovallos F., da Paixão Sevá A., Galucci E., Martins
Soares R., Ferreira F. and Galati E. A. B. “Frequency of Lutzomyia
ID 81-P de Moraes S. C., dos Santos L. H. M., Nunes Alexandre T. and
Brito do Nascimento T. “Sandfly fauna (Diptera: Psychodidae) in
endemic area for transmission of leishmaniasis and the relation of the
presence of Lutzomyia (Lutzomyia) longipalpis and Lutzomyia
(Nyssomyia) whitmani whit information demographics”
ID 82-P Torina A., Reale S., Blanda M., La Russa F., Intile S., Lelli R.,
Vitale F. “Study of the abundance and the distribution of Sandfly in
Sicily”
N° 26 (1) 2015
ID 83-P Berrozpe P., Araujo A., Santini M. S., Utges M. E. and
Salomon O.D. “Lutzomyia longipalpis distribution and abundance in
the city of Corrientes, northeastern Argentina”
ID 84-P de Oliveira Moura Infran J., Alovisi SouzaD., Fernandes de
Souza W., Casaril A. E., Utida Eguchi G., Teruya Oshiro E., Eurico
Fernandes C. and Gutierrez de Oliveira A.“ Study of Phlebotominae
in modified white and black Shannon traps in a Maroon area at
Piraputanga district, Aquidauna, Mato Grosso do Sul, Brazil”
ID 95-P Dutra Rego F., Marteleto Nunes Rugani J., Quaresma P. F.,
Fernandes Shimabukuro P. H., Barbosa Tonelli G., Carvalho Scholte
R. G., and Ferreira Gontijo C. M. “Molecular detection of Leishmania
in sand flies (Diptera: Psychodidae) and spatial distribution of
American Cutaneous Leishmaniasis in the Xakriabá Indigenous
Reserve, Brazil”
ID 85-Pde Oliveira Moura Infran J., Alovisi SouzaD., Fernandes de
Souza w., Casaril A. E., Utida Eguchi G., Elia Teruya Oshiro E.,
Fernandes C. E. and Gutierrez de Oliveira A.“Nycthemeral rhythm
of Phlebotominae in a Maroon area at Piraputanga district,
Aquidauna, Mato Grosso do Sul, Brazil”
ID 86-P Bustamante Gomez M., Diaz M., Espinoza J., Parrado R.,
Reithinger R. and Lineth Garciá A. “Sand Fly Fauna in Chapare,
Bolivia: An Endemic Focus of Leishmania (Viannia) braziliensis”
ID 87-PEspinoza Echeverria J., Bustamante M., Diaz M., Parrado R.
and Lineth Garciá A. “Diversity and nycterohemeral activity of
phlebotomines in San Julian community (Cochabamba, Bolivia)”
ID 88-P Casaril A. E., Falcão de Oliveira E., Zandonaide Monaco N.,
Utida Eguchi G., Rezende de Figueiredo H.,Lopes Fontoura Mateus
N., de Souza Fernandes W., Petilim Gomes S., Alovisi Souza D.,
Oliveira Moura Infran J., Teruya Oshiro E., Galati E. A. B. and
Gutierrez de Oliveira A. “The Sandfly Fauna (Diptera: Psychodidae:
Phlebotominae) in Corumbá, Mato Grosso do Sul, Brazil”
ID 89-Pdos Santos Silva J., Pereira dos Santos A., Kássio Moura
Santos F., Rodrigues da Silva L. O., Caranha L. and Rangel E. F.
“Entomologic surveillance of Phlebotomine sand flies vectors of
Leishmaniasis in areas impacted by the Transfer of the São Francisco
River project, in the state of Ceará, Brazil”
ID 90-P Gebresilassie A., Kirstein O. D., Moncaz A., Tekie H.,
Balkew M., Warburg A., Hailu A., and Gebre-Michael T. “The
Influence of moon light and lunar periodicity on the density of
Phlebotomus (Larroussius) orientalis Parrot, 1936 (Diptera:
Psychodidae) from light trap collections in Ethiopia”
ID 91-PRodríguez-Jiménez J., Pérez-Doria A., Romero-Ricardo L.,
Vergara J., Montesino A., Martínez L., Cortés L., Cochero S., and
Bejarano E. E. “Characteristics of human dwellings and their
surroundings that favour the abundance of phlebotomines in four
urban areas of the Caribbean coast of Colombia”
ID 92-P Barata I. da R., Pinheiro M. S. B., da Silva F. M. M., Feitosa
Brandão R. C., dos Santos T. V. and Silveira F. T. “Phlebotomine sand
fly fauna (Diptera: Psychodidae) and natural flagellate infections
from An area highly endemic for cutaneous leishmaniasis on the
Brazilian-Bolivian frontier in the state of Acre, Brazil”
ID 93-P Franck Thies S., Ribeiro A. L. M., Senatore Rodrigues
Vargas J., Djunko Miyazaki R., Monteiro Michalsky E. and Santos
Dias E. “Sandflies (Diptera: Psychodidae) naturally infected by
Leishmania in rural areas of the city of Nova Mutum, Mato Grosso,
Brasil”
ID 94-P da Silva T. R. R., Gobira Guimarães de Assis M. D., Dutra
Rêgo F., Posteraro Freire M. and Fernandes Shimabukuro P. H.
“Sand fly fauna (Diptera, Psychodidae) collected in the Middle Purus
river region, Amazonas, Brazil”
12
ID 96-P Jomaa Chemkhi, Rihab Yazidi, Mahdi Driss, Jihene Bettaieb
and Afif Ben Salah “First report of natural promastigote infection of
Sergentomyia dreyfussi (Diptera: Psychodidae) sand flies in an
endemic focus of cutaneous leishmaniasis in central Tunisia”
ID 97-P Laurenti M. D., Francesquini F. C., Passero L. F., Tomokane
T. Y., Carvalho A. K., Corbett C. E. P. and Silveira F. T. “Salivary
gland homogenates from wild-caught Lutzomyia flaviscutellata and
L. (Psychodopygus) complexus did not exacerbate Leishmania (L.)
amazonensis and L. (V.) braziliensis infection in BALB/c mice”
ID 98-P Antonio Marque Pereira Júnior, Carolina Bioni Garcia Teles,
Ana Paula de Azevedo dos Santos, Felipe Arley Costa Pessoa, Jansen
Fernandes de Medeiros. "Natural infection of phlebotomine sand flies
(Diptera: Psychodidae) by Leishmania Ross (Kinetoplastida:
Trypanosomatidae) in “terra firme” and “várzea” enviroments in Tefé
municipality, Amazonas state, Brazil”
ID 99-P Martín-Martín I, Jiménez M., González E. and Molina R.
“Transmission of Leishmania infantum to hamsters through the bite of
experimentally infected Phlebotomus perniciosus and monitoring of
animals infectivity by xenodiagnosis”
ID 100-P Bates P.A., Bates M.D. and Kwakye-Nuako G.
“Phylogenetic analysis of the Leishmania enriettii complex and
speculations on their vector biology”
ID 101-P Fernandes M.F., dos Santos K. M., Ferrari Júnior A. C.,
Silva Levay A. P., Negrão F. J., Fernandes W. D., Oshiro E. T.,
Cavalheiros Dorval M. E. M, Andrade Filho J. D., Gutierrez de
Oliveira A. and Galati Eunice A. B. “Vector competence of Nyssomyia
whitmani (Diptera: Psychodidae: Phlebotominae) to Leishmania
(Leishmania) amazonensis”
ID 102-P Rado D., Pita-Pereira D., Araujo Pereira T., Quispe-Florez
MM, Brahim L., Rangel E., Valladares B. and Pacheco R. “Natural
Infection of Lutzomyia (Nyssomyia) y. yuilli and L.
(Trichophoromyia) octavioi with Leishmania spp. in Aguas Calientes,
La Convencion, Cusco, Perú”
ID 103-P Quispe A., Quispe-Ricalde M. A., Rado D., Quispe-Florez
MM, Soto M., Pérez J.E., Valladares B. and Pacheco R. “Lutzomyia
(Nyssomyia) y. yuilli and L. (Trichophoromyia) sp. from Alto
Ivochote, La Convención, Cusco, Peru”
ID 104-P Cutolo A. A, Ovallos F.G., Oliveira C., Lavitschka, Dantas
da Silva M., Molla L. M., dos Santos Nogueira F., Pereira-Chioccola
V. L., Menz I. and Galati E. A. B. “Aggregated pattern of Leishmania
infantum chagasi transmission from naturally infected dogs to
laboratory reared Lutzomyia longipalpis through
xenodiagnosis”
ID 105-P Pech-May Angélica, Sánchez-García Laura, BerzunzaCruz Miriam, Becker-Fauser Ingeborg, Escobedo-Ortegón Javier,
Rebollar-Téllez Eduardo A. " Vectors of Leishmania mexicana in
southeaster of Mexico”
ID 106-P Boussaa Samia,
Ouanaimi Fouad,
Kahime Koloud,
N° 26 (1) 2015
Echchakery Mohamed, Boumezzough Ali and Pesson Bernard
“Genetic characterization of northern and southern populations of
Phlebotomus sergenti (Diptera: Psychodidae) populations in
Morocco”
Pinheiro M. P., da Silva Lopes T. E., de Medeiros Silva M. M. and da
Silva Junior J. B. “Popular knowledge of leishmaniasis transmission
in a peri-urban area of the municipality of São Gonçalo do Amarante,
Brazil”
ID 107-P Noteila M Khalid, Marium A Aboud, Fathi M Alrabba, DiaEldin A Elnaiem and Frederic Tripet. "Evidence for genetic
differentiation at the microgeographic scale in Phlebotomus papatasi
populations from Sudan”
ID 119-P Alves M. de L., Freire de Melo Ximenes M. de F, Fernandes
de Araújo M. F., Gomes Pinheiro M. P., da Silva Lopes T. E., da Silva
Junior J. B. and Gurgel Mendes I.“ Educational comic strip booklet as
an instrument of scientific dissemination of leishmaniasis in the state
of Rio Grande do norte, Brazil”
ID 108-P Oliveira D. M. S., Lima J. A. N., Silva B. J. M., Farias L. H.
S., Santos T.V., Silveira F.T and Silva, E. O. “Analysis of
carbohydrate epitopes on the surface of midgut epithelium of
Lutzomyia (Lutzomyia) longipalpis and Lutzomyia (Nyssomyia)
antunesi (Diptera, Psychodidae) from Pará, State Amazonian
Brazil”
ID 109-P Bussoni Bertollo D. M., de Cassia Hilário Castilho R.,
Costa Nunes Soares M. M., Kindler Figueiredo J., da Silva
Meneghello B. H.; Mitsuyoshi Hiramoto R. and Tolezano J. E.
“Comparative analysis between canine visceral leishmaniasis and
sandfly presence in the endemic municipalities”
ID 110-P Nunes Peres Dias Q., Barbosa de Souza M., Corrêa Novo S.
P., Antônio de Medeiros Meira and do Santo Ponte C. “Survey of
sandfly vectors in areas surrounding Pedra Branca Massif,
Municipality of Rio de Janeiro, Brazil”
ID 111-P Ferrolho J., Gomes J., Alves-Pires C, Cristóvão J.M., Maia
C., Campino L. and Afonso M.O. “Rotation of the external genitalia
in male Phlebotomine sandflies (Diptera, Psychodidae) in laboratory
conditions and in captured specimens in Algarve, Portugal”
ID 112-P Vasconcelos dos Santos T., Batista Campos M., Aprígio
Nunes Lima J., Feitosa Brandão R. C., Medeiros da Silva F. M.,
Barros Pinheiro M. S., da Silva Santos L. A., De Freitas Leão E., da
Rocha Barata I. and Fernando Tobias Silveira “Some field and
laboratory observations on the biology of Nyssomyia antunesi
(Diptera: Psychodiadae), a suspected vector of Leishmania (Viannia)
lindenbergi in Amazonian Brazil”
ID 113-P Goulart T. M., Machado V. E. and Pinto M. C.
“Attractiveness of different diets for Nyssomyia neivai (Diptera:
Psychodidae) larvae”
ID 114-P Goulart T. M., Machado V. E. and Pinto M. C. “Effect of
Nyssomyia neivai (Diptera: Psychodidae) density on
oviposition”
ID 115-P Rita Mukhopadhyay, Goutam Mandal, Jose Orta,
Srotoswoti Mandal, Claudio Meneses, Hamide Aslan Suau, Eric
Beitz, Jesus Valenzuela, Marcelo Ramalho-Ortigao, Shaden
Kamhawi " Role of aquaporins in the physiology of Leishmania
vector Lutzomyia longipalpis”
ID 116-P Dutra Rêgo F., de Souza Pinto I., Machado C. A., Melim
Ferreira G. E., Cupollilo E. and Fernandes Shimabukuro P. H.
“Leishmaniasis Virtual Laboratory (LVL) - a new tool for the study of
American sand flies (Dipetra, Psychodidade, Phlebotominae)”
ID 117-P Mohammad S. Al-Khalifa; Ibraheem M. Al-Hazza and M.
Y. Lubbad “Immunological studies on the cutaneous Leishmaniasis
in Al-Kharj Region, Saudi Arabia”
ID 118-P Alves M. de L., Freire de Melo Ximenes M. de F., Gomes
13
ID 120-P Gomes Pinheiro M. P., Freire de Melo Ximenes M. de F., de
Melo Pereira Pinheiro M. P., de Lima Alves M. “Cordel folk literature
as a tool for preventing leishmaniasis in Brazil”
ID 121-P Rea M. J. F., C Borda. E. “Survey of American
tegumentary leishmaniasis in the northeast of Argentina during 20112014”
ID 122-P Lozano A., Basmadjián Y., Vitale E., Satragno D., Canneva
B., Verger L., Tort C., Viera A., Romero S., Ríos C., Lagarmilla P.
"Canine Visceral Leishmaniasis. Imported cases in Uruguay, 20102014"
ID 123-P de Freitas Rocha A. T., Melo Silva S., Araújo Soares M. R.,
Costa Fortier D., Silva Barbosa D. B., Nery Costa C. H. " Occurrence
of Lutzomyia longipalpis and spatial distribution of visceral
leishmaniasis in Floriano, Piauí, Brazil: preliminary data”
ID 124-P Santos M. F. C., Paranhos Filho A. C., Gamarra R. M.,
Fernandes W. D.; Brazil R. P. and Oliveira A. G. “Use of Remote
Sensing in the analysis of populations of Lutzomyia longipalpis
complex”
ID 125-P Utgés M. E., Fuenzalida D., Parras M., Gould I. T.,
Casertano, S. and Salomón O. D. “Geometric morphometry of
northeastern populations of Nyssomyia neivai from Argentina”
ID 126-P Rado D., Pérez J. E., Vilela M., Quispe D., Quispe A.,
Aguilar E. G., Rangel E., Valladares B., Pacheco R. “Sandflies
(Diptera: Psychodidae) of Aguas Calientes, La Convención, Cusco,
Perú”
ID 127-P Toccas F., Chacon M., Pérez J. E., Rado D., Soto M.,
Mendoza J., Luna R., Aguilar E. G., Valladares B. and Pacheco R.
“Lutzomyia spp. (Diptera: Psychodidae) in intra, peri and
extradomiciliar environments in Yomentoni (right margin), La
Convención, Cusco, Perú”
ID 128-P Rezende de Figueiredo H., Casaril A. E., Infran J. O. M.,
Teruya Oshiro E., Rodrigues Mendes A., Andrade Filho J. D., Moraes
Ribeiro L., Falcão Oliveira E., da Cunha Santos M. F. and Gutierrez de
Oliveira A. “Ecology of Lutzomyia longipalpis (Diptera:
Psychodidae) in Aquidauana, endemic area for leishmaniases, Mato
Grosso do Sul, Brazil. Preliminary results”
ID 129-P Antonio Marques Pereira Júnior, Eric Fabricio Marialva,
Moreno de Souza Rodrigues, Felipe Arley Costa Pessoa, Jansen
Fernandes de Medeiros. " Studies on fauna of phlebotomine sand flies
(Diptera: Psychodidae) at “Terra firme” and “Várzea” environments
in Tefé municipality, Amazonas state, Brazil”
ID 130-P Jiménez M., González E., Hernández S. and Molina R.
“Phlebotomine sand fly survey in the focus of human leishmaniasis in
south western Madrid region, Spain: 2012-2013”
N° 26 (1) 2015
ID 131-P Jiménez M., González E., Hernández S. and Molina R.
“Molecular detection of Leishmania infantum and host-feeding
preferences in Phlebotomus perniciosus from the focus of
leishmaniasis in south western Madrid region, Spain: 20122013”
P. C., de Almeida M. R. “Sandflies (Diptera, Psychodidae,
Phlebotominae) in Pitoco cave, an archeological site in northern Mato
Grosso do Sul state, Brazil”
ID 132-P Zoghlami Z., Chouihi E., Barhoumi W., Dachraoui K.,
Massoudi N., Ben Helel K., Habboul Z., Hadhri§ M. H., Limam S.,
Mhadhbi M., Gharbi M., and Zhioua E. “Interaction between canine
and human visceral leishmaniases in a holoendemic focus of Central
Tunisia”
ID 133-P Nilsa Gonzalez-Britez, Nilda Portillo, Maria Ferreira,
Martha Torales, Nidia Martínez, Luciano Franco. "Leishmaniasis
vectors in the Alto Paraná Department, Paraguay”
ID 134-P da Silva D. F., de Castilho Sanguinette C., Barbosa Tonelli
G., Tanure A., Vidal Stumpp R. G. A, Dutra Rêgo F., Gontijo C. M. F.
and do Andrade Filho J. D. “Sand flies (Diptera: Psychodidae:
Phlebotominae) collected in urban and wild area in the district of
Barra do Guaicui, Várzea da Palma municipality, Minas Gerais,
Brazil”
14
ID 144-P Santos Carvalho S. M., Fitipaldi Veloso Guimarães V. C.,
dos Santos P. R. B. “First report of Lutzomyia longipalpis in Uruçuca,
cocoa zone of the southern state Bahia, Brazil”
ID 145-P Ballart C., González E., Ravel C., Jiménez M., Abras A.,
Molina R., Portús M., Gállego M. “Identification of blood meals in
Leishmania infantum vectors from the Northeast of Spain:
Phlebotomus ariasi and P. perniciosus by Polymerase Chain Reaction
based and sequencing methods”
ID 146-P de Lima Alves M., de Melo Ximenes M. F. F., Pinheiro M. P.
G., da Silva J. H. T., da Silva Lopes T. E., de Medeiros Silva M. M.
“Phlebotomine sanflies (Diptera: Psychodidae) in a peri-urban setting
in the metropolitan region of Natal, Brazil”
ID 147-P Toccas F., Rado D., Quispe A., Quispe C., Galindo B.,
Chacon M., Quispe-Ricalde MA, Valladares B., Pacheco R. “The
Sand Fly Fauna (Diptera: Psychodidae) in Alto Ivochote, La
Conveción, Cusco, Perú”
ID 135-P Neves Vianna E., Franco Morais M. H., Sobral de Almeida
A., Chagastelles Sabroza P., Carneiro M. and Santos Dias E.
“Evaluation of the abundance of phlebotomine sand flies in
households from an area of Visceral Leishmaniasis transmission in
Belo Horizonte, Minas Gerais, Brazil”
ID 148-P Martínez M. F., Santini M. S., Utgés M. E., Kowalewski M.,
Salomón O. D. “Study of phlebotomines in two wild areas of
northeastern Argentina and the potential association with howler
monkeys (Alouatta caraya)”
ID 136-P Cavalcante Lamy Serra e Meira P., Lacerda Abreu B.,
Lusardo de Almeida Zenóbio A. P., de Castilho Sanguinette C., Mayr
de Lima Carvalho G., Saraiva L. and Andrade Filho J. D.
“Phlebotominae sand fly fauna in two urban parks in Belo Horizonte,
Minas Gerais, Brazil (Diptera: Psychodidae)”
ID 149-P de Melo Ximenes M. F. F., Antonaci Gama R., da Silva
Lopes T. E., Melo Freire R. C., de Lima Alves M., Pinheiro M. P. G.,
da Silva J. H. T., de Medeiros Silva M. M. “Phlebotominae sanflies
(Diptera: Psychodidae) in a preserved area of the Caatinga biome in
Rio Grande do Norte State, Brazil”
ID 137-P Cavalcante Lamy Serra e Meira P. and Andrade Filho J. D.
“Species of phlebotomine sand flies (Diptera: Psychodidae:
Phlebotominae) reported in Brazilian caves”
ID 150-P Basmadjián Y., Canneva B., Verger L., Vitale E., Lozano
A., Sequeira C., Lozano A., Satragno D., Tort C., Viera A., Ríos C.,
Lagarmilla P. and Romero S. “Lutzomyia longipalpis in Bella Unión,
Department of Artigas, Uruguay. One year of surveillance: July
2013- June 2014”
ID 151-P Oliveira A.F.J., Freitas R.A., Pessoa F.A.C., Aguiar N.O.
“Phlebotomine Sand Fly survey in Central Amazon, Amazonas State,
Brazil with a new register of Lutzomyia caligata”
ID 138-P Barbosa Tonelli G., Dutra Rego F., Mayr de Lima Carvalho
G., Tanure A. and Andrade Filho J. D. “Ecology of sand flies (Diptera:
Psychodidae: Phlebotominae) in the Natural Heritage Private
Reserve of Serra do Caraça, Minas Gerais, Brazil”
ID 139-P Tanure A., Barbosa Tonelli G., Dutra Rêgo F., Mayr de Lima
Carvalho G., Gontijo C. M. F., Fontes Paz G. and Andrade Filho J. D.
“Study of sand flies (Diptera: Psychodidae: Phlebotominae) in a
transmission area of leishmaniasis in Minas Gerais, Brazil”
ID 140-P Souza G. D., A. Cardoso, de Carvalho R. M. J. S.
“Assessment of sand flies (Diptera: Psychodidae) in an area with
canine visceral leishmaniasis in the municipality of Porto Alegre, Rio
Grande do Sul, Brazil”
ID 141-P Basmadjián Y., Canneva B., Verger L., Lozano A.,
Satragno D., Supparo E., Sequeira C., Romero S., Viera A., Tort C.,
Ríos C., Lagarmilla P. and Vitale E. “Persistence of Lutzomyia
longipalpis, vector of Visceral Leishmaniasis, in the city of Salto,
Uruguay”
ID 142-P Verger L, Lozano A, Vitale E, Satragno D, Sequeira C,
Canneva B, Basmadjián Y, Rios C, Lagarmilla P, Tort C, Viera A,
Romero S. “Serological surveillance in dogs of the city of Bella
Unión, a risk area for Visceral Leishamaniasis”
ID 143-P Galati E. A. B., de Almeida P. S., de Paula M. B., Costa Filho
ID 152-P de Arruda Moura A. C., da Silva K. G., D. de Oliveira
Miranda E., Ramalho M., da Silva F. J., Brandão-Filho S. P., Otranto
D., Dantas-Torres F. “Phlebotominae sanflies in military training area
in north-eastern Brazil”
ID 153-P Ferro Cr., Lopez M., Fuya P., Lugo L., Cordovez J. M.,
Gonzalez C. “Relating vector species potential distribution and
richness to transmission foci of Cutaneous Leishmaniasis in
Colombia”
ID 154-P Ballart C., Pesson B., Martín-Sánchez J., Alves-Pires C.,
Morillas F., Afonso O., Portús M., Gállego M. “Risk of leishmaniosis
transmission in the Pyrenean region of Spain assessed through the
isoenzymatic analysis of potential sand fly vectors (Phlebotomus
ariasi and P. perniciosus)”
ID 155-P Barhoumi W., Qualls W. A., Archer R., Fuller D. O., Chelbi
I., Cherni S., Derbali M., Arheart K. L., Zhioua E.and Beier J. C.
“Irrigation in the arid regions of Tunisia impacts the abundance and
apparent density of sand fly vectors of Leishmania infantum”
N° 26 (1) 2015
ID 156-P Pinheiro M. P. G., de Melo Ximenes M. F. F., de Lima Alves
M., da Silva Júnior J. B., da Silva J. H. T., M de Medeiros Silva. M., da
Silva Lopes T. E. “ Sandflies (Diptera: Psychodidae: Phlebotominae)
from the Nisia Floresta National forest, metropolitan region Natal,
Rio Grande do Norte State, Brazil”
ID 170-P Erin Dilger, Orin Courtenay and Graham Medley “Density
dependence in host-vector relationships and consequences for
transmission”
ID 157-P Erisoz Kasap O., Alten B. “Entomological Survey of
Phlebotomine Sand Flies (Diptera: Psychodidae) in Turkey”
ID 158-P Copa G. N., Marco J. D., Nasser J. R., Gil J. F. “Distribution
of sandflies species in a locality endemic for American tegumentary
leishmaniasis from the north of Argentina, bordering with Bolivia”
ID 159-P Ruiz-Lopez F., Cadena H., Vélez Mira A., Posada-López L.,
Contreras A., Tirado Duarte D., Vélez I. D. “Effectiveness of a rapid
multidisciplinary protocol for the epidemiological evaluation of an
active focus of cutaneous leishmaniasis: Remedios, Antioquia,
Colombia”
ID 160-P Villarquide M. L., Fernández M. S., Santini M. S., Salomón
O. D. “Abundance Association of Nyssomyia whitmani with the
number of farm animals and its dwelling distance to the house, in
areas with Tegumentary Leishmaniasis transmission in Puerto
Iguazú, Misiones, Argentina”
ID 161-P Ferro C., Fuya P., Lugo L., González C. “Biodiversity of
Phlebotominae subfamily (Diptera: Psychodidae) in the Colombian
Amazon and Orinoco”
ID 162-P Enriquez S., Ariviliaga J., Vaca F., Benitez Ortiz W.,
Kirstein O., Abassi I., M Calvopiña., Pozo K., Romero V., O
Chiluisa., E Villacrés and Warburg A. “Sand fly vectors of cutaneous
leishmaniasis on the Western slopes of the Ecuadorian Andes”
ID 163-P Direni Mancini J. M., Fuenzalida A. D., Saracho Bottero M.
N., Lizarralde de Grosso M. S., Salomón O. D., Quintana M. G.
“Abundance of Phlebotominae in endemic area of cutaneous
leishmaniasis, Tucumán, Argentina”
ID 164-P Ouanaimi F., Boussaa S., Kahime K., Echchakery M.,
Boumezzough A. “Phlebotomine sandflies (Diptera: Psychodidae) of
Morocco: Results of entomological surveys along the OuarzazatM'hamid, Zagora-Chefchaouen-Nador and Zagora-Tata-Marrakesh
roads”
ID 165-P Kahime K., Boussaa S., Ouanaimi F. and Boumezzough A.
“Spatial relationship between environmental factors and sand fly
(Diptera: Psychodidae) distribution associated with leishmaniasis
risk in centre & southern Morocco”
ID 166-P Boussaa S., Ouanaimi F., Boumezzough A. “Identification
and characterization of larval lodging of phlebotomine sand flies
(Diptera: Psychodidae) in Marrakech, Morocco”
ID 167-P Boumezzough A., Kahime K., Boussaa S. and Ouanaimi F.
“Leishmaniasis in Morocco: still a topical question”
ID 168-P Remondegui C. V., Cabrera C. H. A., Quintana M. G.
“Seasonal fluctuation of Phlebotominae (Diptera: Psychodidae) in
three Departments of an endemic area of Cutaneous Leishmaniasis in
Jujuy province. A preliminary report”
ID 169-P Bisetto Júnior A., Bisetto M., Dotto Ampessam S. M.
“Spatial analyisis of the accurrence of leishmaniasis in Paraná, Brazil,
to identify areas of risk”
15
ID 171-P Casanova C., Fernandes Shimabukuro P. H., Almeida
Bressiani B., Fernanda Lazari T., Galati E. A. B. “Geographical
distribution of Nyssomyia neivai (Pinto) and Nyssomyia intermedia
(Lutz & Neiva) in the state of São Paulo, Brazil”
ID 172-P Bergo E., Nascimento L., Camargo-Neves V. “First meeting
of Lutzomyia longipalpis in rural areas related to outbreak of canine
visceral leishmaniasis in the northwestern region of São Paulo,
Brazil”
ID 173-P Colla-Jacques F. E., Casanova C., Galvis Ovallos F. “Study
of sand fly community in two rural endemic areas for American
cutaneous leishmaniasis in the municipalities of Mogi Guaçu and
Mogi Mirim, São Paulo State, Brazil”
ID 174-P Saraiva L, Rugani J. M. N., Lima A.C.V. M. R., Reis A.S.,
Pereira A. A. S., Gontijo C.M.F., Andrade Filho J.D. “DNA detection
of Leishmania sp. in female sand flies (Diptera: Psychodidae) in Belo
Horizonte city and Sumidouro State Park, Minas Gerais – Brazil”
ID 175-P Santini M. S., Utges M. E., Casas N., Salomon O. D.
“Association Of Occurrence And Abundance Of Lutzomiya
longipalpis with Environmental Variables At Different Spatial Scales:
Micro And Macro-Scale”
ID 176-P Waleed Al Salem, Badriah Al Otaibi, Louise Kelly-Hope,
Karina Mondragon-Shem, Salah Balgonaeem, Maha Abdeladhim,
Mohammed Al Zahrani, Amir Hassan, Jesus Valenzuela Alvaro
Acosta-Serrano “A Public Health Approach to Leishmaniasis Control”
INDEX BY TOPIC
Vector Control
1-O, 2-O, 3-O, 4-O, 5-O, 6-O, 7-O, 8-O, 9-O, 10-O, 11-O, 12-O
1-P, 2-P, 3-P, 4-P, 5-P, 6-P, 7-P, 8-P, 9-P, 10-P, 11-P, 176-P
Taxonomy & Phylogeny
13-O, 14-O, 15-O, 16-O, 17-O, 18-O, 19-O, 20-O, 21-O, 22-O, 23-O,
24-O
47-P, 48-P, 49-P, 50-P, 51-P, 52-P, 53-P, 54-P, 55-P, 56-P, 57-P, 58-P,
59-P, 60-P, 61-P, 62-P, 63-P, 64-P, 65-P, 66-P
Ecology & Epidemiology
25-O, 26-O, 27-O, 28-O, 29-O, 30-O, 31-O, 32-O, 33-O, 34-O, 35-O,
36-O, 37-O, 38-O, 39-O, 40-O, 41-O, 42-O. 43-O, 44-O, 45-O
12-P, 13-P, 14-P, 15-P, 16-P, 17-P, 18-P, 19-P, 20-P, 21-P, 22-P, 23-P,
24-P, 25-P, 26-P, 27-P, 28-P, 29-P, 30-P, 31-P, 32-P, 33-P, 34-P, 35-P,
36-P, 37-P, 38-P, 39-P, 40-P, 41-P, 42-P, 43-P, 44-P, 45-P, 46-P, 67-P,
68-P, 69-P, 70-P, 71-P, 72-P, 73-P, 74-P, 75-P, 76-P, 77-P, 78-P, 79-P,
80-P, 81-P, 82-P, 83-P, 84-P, 85-P, 86-P, 87-P, 88-P, 89-P, 90-P, 91-P,
123-P, 124-P, 125-P, 126-P, 127-P, 128-P, 129-P, 130-P, 131-P, 132-P,
133-P, 134-P, 135-P, 136-P, 137-P, 138-P, 139-P, 140-P, 141-P, 142-P,
143-P, 144-P, 145-P, 146-P, 147-P, 148-P, 149-P, 150-P, 151-P, 152-P,
153-P, 154-P, 155-P, 156-P, 157-P, 158-P, 159-P, 160-P, 161-P, 162-P,
163-P, 164-P, 165-P, 166-P, 167-P, 168-P, 169-P, 170-P, 171-P, 172-P,
N° 26 (1) 2015
16
173-P, 174-P, 175-P
Vector Competence & Infections
46-O, 47-O, 49-O, 50-O, 51-O, 52-O
92-P, 92-P, 93-P, 94-P, 95-P, 96-P, 97-P, 98-P, 99-P, 100-P, 102-P, 102P, 103-P, 104-P, 105-P
Leishmania – Sandfly /host Interactions
48-O, 53-O, 54-O, 55-O, 56-O, 57-O, 58-O, 59-O, 60-O
108-P, 109-P
Genome & Genetics/ Virus infections
61-O, 62-O, 63-O, 64-O, 65-O, 66-O, 67-O, 68-O, 69-O
106-P, 107-P, 109-P, 110-P
Physiology & Colonization
70-O, 71-O, 72-O, 73-O
111-P, 112-P, 113-P, 114-P, 115-P
Other Topics
116-P, 117-P, 118-P, 119-P, 120-P, 121-P, 122-P
INDEX BY AUTHOR (SURNAME)
A
Abassi Ibrahim 36-O, 34-P, 41-P, 162-P
Abdeladhim Maha 8-O, 28-O, 112-O,
176-P
Abdeliyev Ziyat 9-P
Aboud Marium A 66-O
Abras A 145-P
Abreu Bruna Lacerda 136-P
Abril Marcelo 44-P
Acardi Soraya Alejandra 3-P, 45-P
Acel Anagela Maria 26-P, 27-P, 28-P
Acosta Luz A 26-O, 8-P, 63-P
Acosta M Manteca 11-O
Acosta-Serrano Alvaro 8-O, 28-O, 176-P
Afonso Margarete Martins dos Santos 18-P,
19-P
Afonso MO 33-P, 57-P, 111-P, 154-P
Aguiar NO 151-P
Aguilar E 27-O, 126-P, 127-P
Aimakhanov Batirbek 9-P
Akhavan AA 40-O
Aklilu Esayas 37-O
Alexandre Thiago Nunes 81-P
Al-Hazza Ibraheem M 117-P
Alkan C 39-O
Al-Khalifa Mohammad S 117-P
Almazán María Cristina 67-P
Al Otaibi Badriah 112-O, 176-P
Alrabba Fathi M 66-O
Al Salem Waleed 28-O, 8-O, 176-P
Alten B 21-O, 39-O, 157-P
Alves Leucio Câmara 31-P
Alves Maria de Lima 118-P, 119-P, 120-P,
146-P, 149-P, 156-P
Alves-Pires C 111-P, 154-P
Alves Veracilda Ribeiro 20-P, 22-P, 55-P
Al Zahrani Mohamed 8-O, 28-O, 176-P
Andrade-Filho José Dilermando 65-O, 13P, 25-P, 58-P, 60-P, 66-P, 72-P, 101-P, 128-P,
134-P, 136-P, 137-P, 139-P, 174-P
Alzogaray Raul A 5-P
Amini Esfanjani MH 61-P
Ampessam Sônia Maria Dotto 169-P
Añez Néstor 25-O
Arandian MH 40-O
Araujo Analia V 38-P, 83-P
Archer Reginald 155-P
Arcones Carolina 17-O, 22-O
Arheart Kristopher L 155-P
Arivillaga Jazzmin 162-P
Arserim Suha K 4-O, 36-P, 37-P
Atshabar Bakhit 9-P
Austin James 1-O
Aydın S 39-O
B
Balcioğlu I Cüneyt 37-P, 4-O
Balgonaeem Salah 8-O, 176-P
Balkew Meshesha 35-O, 36-O, 37-O, 38-O,
29-P, 34-P, 90-P
Ballart C 145-P, 154-P
Baneth Gad 73-P
Banuls A-L 39-O
Barata Iorlando da Rocha 92-P, 112-P
Barazesh A 61-P
Barbosa Diogo Brunno e Silva 123-P
Bargues María Dolores 18-O
Barhoumi W 3-O, 44-O, 132-P, 155-P
Barresi Trey 2-O
Barroso Paola A 67-P
Basano Fabrizio Solari 12-O
Basmadjián Yester 122-P, 141-P, 142-P,
150-P
Bates MD 100-P
Bates Paul A 50-O, 55-O, 100-P
Batista Hebert Lima 78-P
Becker-Fauser Ingeborg 105-P
Beier John C 155-P
Beitz Eric 115-P
Bejarano Eduar Elías 26-O, 46-O, 51-O, 8P, 59-P, 63-P, 91-P
Belen A 39-O
Bell Melissa 46-P
Ben Helel K 132-P
Ben Salah Afif 49-P, 96-P
Bénié Goze Bertin 32-O
Benítez Ibalo P 38-P
Bergo Eduardo 172-P
Berrozpe Pablo 83-P
Bertollo Denise Maria Bussoni 109-P
Berzuna-Cruz Miriam 105-P
Bettaieb Jihene 32-O, 96-P
Bettiol Flávia Karolina Pereira Barreto 27P, 28-P
Bevilaqua Claudia ML 16-P
Bevilacqua Paula Dias 72-P, 60-P
Bianchi R 68-O
Bichaud L 44-O
Bisetto Júnior Alceu 11-P, 169-P
Bisetto Maurício 169-P
Blanda M 82-P
Bongiorrno G 45-O, 68-O
Borda Carlos Edgardo 62-P, 121-P,
Borges-Alves G 14-O
Botelho Patrick Feitosa 54-P
Boujaâma A 3-O
Boumezzough Ali 106-P, 164-P, 165-P,
166-P, 167-P
Boussaa Samia 106-P, 164-P, 165-P, 166-P,
167-P
Brahim L 102-P
Brandão-Filho Sinval Pinto 12-P, 30-P, 31P, 32-P, 152-P
Brandão Roberto Carlos Feitosa 92-P, 112P
N° 26 (1) 2015
Bray DP 14-O
Brazil Reginaldo Peçanha 14-O, 60-P, 72P, 124-P
Bressiani Bruna Almeida 171-P
Brianti Emanuele 12-O
Brilhante Andreia Fernandes 17-P
Browers Marinês 2-P
Burleigh Barbara 69-O
Burruss Dylan 2-O, 3-O
C
Cabrera CHA 168-P
Cabrera Olga Lucía 7-O, 9-O, 10-O
Cáceres Abraham G 15-O
Cadavid Gloria 73-O
Cadena Horacio 26-O, 8-P, 63-P, 159-P
Caligiuri Lorena G 3-P
Calvopiña Manuel 162-P
Calzada Jose E 35-P
Camargo-Neves Vera 172-P
Cameron Mary 46-P
Campino L 33-P, 57-P, 111-P
Campos Marliane Batista 112-P
Canneva Bruno 122-P, 141-P, 142-P, 150P
Caranha Lindemberg 89-P
Cardoso A 140-P
Cardoso Cristiane de Oliveira 17-P
Carneiro Mariângela 135-P
Carpenter S 50-O, 143-O
Carter V 14-O
Carvalho AK 97-P
Carvalho Gustavo Mayr de Lima 25-P,
136-P, 139-P
Carvalho Silvia María Santos 144-P
Casanova Claudio 171-P, 173-P
Casaril Aline Etelvina 84-P, 85-P, 88-P,
128-P
Casertano Sergio A 58-P, 125-P
Castilho Regiane de Cassia Hilário 109-P
Castillo G 67-P
Cavalcanti Milena de Paiva 32-P
Cavia Regino 21-P
Çetin Hüseyin 4-O
Chacon M 27-O, 127-P, 147-P
Chagas Anadeiva Portela 30-O
Chalghaf Bilel 32-O
Chanampa Mariana M 67-P
Charrel RN 39-O, 44-O
Chaves Sérgio Augusto de Miranda 18-P,
19-P
Chavez Luis F 35-P
Checa Rocío 22-O, 17-O
Chemkhi Jomaa 49-P, 96-P
Chelbi Ifhem 3-O, 44-O, 155-P
Cherni Saifedine 3-O, 44-O, 155-P
Chicharro Cármen 17-O, 22-O
Chiluisa Orlando 162-P
Chlebi Ifhem 72-O
Chlif Sadok 32-O
Chouihi E 132-P
Ciufolini MG 68-O
Cochero Suljey 46-O, 51-O, 91-P,
Coelho Tháis Oliveira 6-P
Colla-Jacques Fernanda Elisa 173-P
Collins Frank H 62-O
Contreras Angélica 159-P
Copa Griselda N 67-P, 158-P
Corbett CEP 97-P
Cordeiro Jose LP 42-P
Cordovez Juan Manuel 153-P
Corrêa Arlene Gonçalves 71-O
Cortés Luis 51-O, 91-P
Cosenza M 23-O
Costa Carlos Henrique Nery 10-P, 123-P
Costa Dorcas L 10-P
Costa Filho Pedro Catarino 143-P
Costa Pietra Lemos 12-P, 30-P
Costa Simone M 18-P
Courtenay O 14-O, 170-P
Cristóvão JM 33-P, 57-P, 111-P
Cruz Israel 17-O, 22-O
Cruz Susana 22-O
Cumbrera Anayansi Valderrama 65-O
Cupollilo Elisa 116-P
Cutolo Andre Antonio 104-P
D
Dachraoui K 44-O, 132-P
da Costa Ailton Junior Antunes 74-P, 75-P
da Costa Simone Miranda 19-P, 42-P
da Fonseca Claudio Ruy V 55-P
D'Agostino R 23-O
Dagur Pradeep K 56-O
Damazo Amilcar Sabino 1-P
Dantas-Torres Filipe 12-P, 30-P,31-P, 32-P,
152-P, 12-O
Darabi H 40-O
Darvishi M 40-O
Da Paixão Fabiana Rodrigues 75-P
da Silva Ana de Nazaré Martins 76-P
da Silva Andrea Martins 69-O
da Silva Danyele Franca 134-P
da Silva Fábio Márcio Medeiros 54-P, 92-P,
112P
da Silva Fernando José 12-P, 30-P, 152-P
da Silva Jalthon C 10-P
da Silva José Hilário Tavares 146-P, 149-P,
156-P
da Silva Junior João Batista 118-P, 119-P,
156-P
da Silva Kamila Gaudêncio 31-P, 32-P,
152-P
da Silva Luiz Osvaldo Rodrigues 89-P
17
Da Silva Maria Elisabel Tavares 77-P
da Silva Mariana Dantas 104-P
da Silva Shara Regina 74-P
da Silva Túllio Romão R 94-P
Das Pradeep 6-O
Davidson Silas 7-P
De Almeida Andréa Sobral 135-P
de Almeida Paulo Silva 29-O, 143-P
de Almeida Romoaldo Martins 143-P
de Almeida Tiago Parreiras 27-P, 28-P
de Andrade Andrey José 13-O, 29-O, 6-P,
14-P, 50-P, 52-P, 56-P
de Araújo Magnólia Florêncio Fernandes
119-P
de Araújo Patrick Thiago Carvalho 76-P
de Assis Mauro Diego Gobira Guimarães
94-P
de Avelar Daniel Moreira 74-P
de Carvalho Bruno M 18-P, 19-P, 40-P
de Carvalho Gílcia Aparecida 31-P
de Carvalho Maria do Socorro Laurentino
29-O
de Carvalho RMJS 140-P
de Figueiredo Helen Rezende 88-P, 128-P
De Lazzari Elisa 1-O
de Medeiros Jansen Fernandes 98-P, 129-P,
de Melo Fábio Lopes 12-P
Demir Samiye 4-O
de Moraes Sinara Cristina 80-P, 81-P
de Morais Rayana Carla Silva 32-P
de Oliveira Alexandra Gutierrez 84-P, 85-P,
88-P, 101-P, 128-P
de Oliveira Everton Falcão 88-P
Depaquit Jerome 16-O, 18-O, 20-O, 21-O,
24-O, 64-O
de Paula Marcia Bicudo 143-P
de Pita Pereira Daniela 19-P
Derbali Mohamed 3-O, 44-O, 155-P
Derrick Tamsyn 55-O
de Santana Antônio Luis Ferreira 19-P
de Souza Emerson Marques 27-P, 28-P
de Sousa Gilberto César 30-O
de Souza Jailson Ferreira 17-P
de Souza Marcos Barbosa 110-P
de Souza, Nataly A 48-P
de Souza Wagner Fernnades 84-P, 85-P
Deuster Katrin 12-O
Dey Ranadhir 56-O
Dias Edelberto Santos 1-P, 74-P, 75-P, 79-P,
93-P, 135-P
Dias Quezia Nunes Peres 110-P
Dias TA 25-P
Diaz-Albiter Hector M 67-O
Diaz Mery 86-P, 87-P
Di-Blasi Tatiana 54-O, 59-O
Dilger E 14-O, 170-P
Dillon Roderick J 55-O, 67-O
N° 26 (1) 2015
Di Muccio T 45-O
Dinesh Diwakar Singh 6-O
Direni-Mancini JM 65-P, 163-P
Donato Lucas Edel 16-P
do Nascimento Thais Brito 81-P
Dorval Maria Elizabeth Moraes
Cavalheiros 101-P
dos Santos Ana Paula de Azevedo 98-P
dos Santos Antonio Pereira 89-P
Dos Santos Carlos Fernando Rocha 15-P,
77-P
dos Santos Eric Fabricio Marialva 129-P
dos Santos Fabio Alexandre Leal 27-P
dos Santos Klieton Maciel 101-P
dos Santos Lucas Heber Mariano 81-P
dos Santos Nathanna Progênio 17-P
dos Samtos Paulo Raimundo Barbosa 144P
dos Santos Thiago Vasconcelos 48-P, 54-P,
76-P, 92-P, 112-P
dos Santos Yago Costa Vasconcelos 76-P
Driss Mahdi 96-P
Duarte Didier Tirado 159-P
Duffy Darragh 57-O
Duncan Robert 56-O
D'Urso V 45-O
Dvořák Vít 19-O, 20-O, 21-O, 39-O
E
Echchakery Mohamed 106-P, 164-P,
Echeverría Jorge Espinoza 86-P, 87-P
Eguchi Gabriel Utida 84-P, 85-P, 88-P
Elnaiem Dia-Eldin A 31-O, 66-O
Emrich Scott J 62-O
Enriquez Sandra 162-P
Ergunay K 39-O
Ermis V Ozge 37-P
Escobedo-Ortegón Javier 105-P
Espada Liz 64-P
Estrada Luis G 26-O, 8-P, 63-P
F
Fadul Mônica 76-P
Faiman Roy 39-P
Fares W 44-O
Faria Maiara d 67-O
Farias LHS108-P
Farias Teresinha C 10-P
Faustino Maria Aparecida da Gloria 31-P
Fernándes Carlos Eurico 84-P, 85-P
Fernandes Magda Freitas 101-P
Fernandes Wedson Desidério 101-P, 124-P
Fernández María Soledad 21-P, 43-P, 160-P
Fernández Roberto 64-P
Fernández Wagner de Souza 88-P
Ferrari Júnior Antonio Carlos 101-P
Ferreira Fernando 70-P, 71-P
Ferreira Gabriel Eduardo Melim 116-P
Ferreira Jônatas Barbosa Cavalcante 50-P
Ferreira Maria 133-P
Ferro Cristina 153-P, 161-P
Ferrolho J 111-P
Figueiredo Juliana Kindler 109-P
Fiuza Vanessa de Oliveira Pires 74-P
Fortes-Dias Consuelo Latorre 74-P, 79-P
Fortier Daniel Costa 123-P
Fortuna C 68-O
França-Silva João Carlos 75-P
Francesquini FC 97-P
Franco Luciano 133-P
Freire Maíra Posteraro 20-P, 94-P
Freire Renato Cesar Melo 149-P
Freitas FB 33-P
Freitas RA 151-P
Freitas Vanesa C 48-O
Fuenzalida A Denise 65-P, 125-P, 163-P
Fuller Douglas O 155-P
Fux Blima 13-P
Fuya Patricia 153P, 161-P
G
Galati Eunice Aparecida Bianchi 13-O, 17P, 48-P, 51-P, 52-P, 56-P, 70-P, 71-P, 88-P,
101-P, 104-P, 143-P, 171-P
Galindo B 147-P
Gállego M 145-P, 154-P
Gálvez Rosa 17-O, 22-O
Galucci Estela 70-P, 71-P
Gama Renata Antonaci 149-P
Gamarra Roberto M 124-P
Garcez Lourdes María 30-O
Garciá Ana Lineth 86P, 87-P
Garlapati Rajesh B 2-O
Garrido Renata 18-P
Gaudêncio Kamila 30-P
Gebre-Michael Teshome 33-O, 34-O, 35O, 36-O, 37-O, 38-O, 29-P, 34-P, 39-P, 41P, 90-P
Gebresellassie Araya 33-O, 34-O, 36-O,
39-P, 41-P, 90-P
Gendernalik Alex 9-P
Genta Fernando A 67-O
Gharbi M 132-P
Gil José 67-P, 158-P
Giordano Cristina 19-P
Giraldo-Calderón Gloria I 62-O, 63-O
Giraud Emile 55-O
Godoy Rodrigo Espíndola 48-P, 54-P
Gomes J. ID 111-P
Gomes Suellem Petilim 88-P
Gómez Bravo Andrea 44-P
Gómez-Bustamante Marinely 86-P, 87-P
Gómez Hernando 8-P
Gonçalves Raquel 30-O
18
Gonçalves Rodrigo Gurgel 6-P
Gontijo Célia Maria Ferreira 25-P, 66-P, 95P, 134-P, 139-P, 174-P
González-Britez Nilsa 11-P, 133-P
González Camila 153-P, 161-P
González E 145-P
González Estela 99-P, 130-P, 131-P
Goulart Thais Marchi 71-O, 113-P, 114-P
Gould Ignacio Tomás 5-P, 43-P, 44-P, 125-P
Gradoni Luigi 73-P, 45-O
Gramiccia Marina 73-P, 45-O, 68-O
Grant William E 5-O
Graser Carina 18-P
Guerbouj Souheila 49-P
Guimarães Ricardo 30-O
Guimarães Vanessa Cristina Fitipaldi
Veloso 12-P, 30-P, 144-P
Guizani Ikram 49-P
Gunay 39-O
Gurgel-Gonçalves Rodrigo 29-O, 14-P, 50P,
H
Habboul Z 132-P
Hadhri MH 132-P
Hailu Asrat 19-O, 33-O, 34-O, 29-P, 34-P,
35-O, 36-O, 37-O, 38-O, 39-P, 41-P, 73-P,
90-P Halada Petr 20-O
Hamida Nabil Belhadj 58-O
Hamilton J Gordon C 14-O, 72-O
Harrabi Mariem 32-O
Hasan Milena 57-O
Hashiguchi Yoshihisa 15-O
Hassan Amir 112-O, 176-P
Hassan Faizan 6-O
Hawksworth Mallory 63-O
Hermes Sandra Cristina Negreli Moreira
29-O
Hernández Sonia 130-P, 131-P
Hiramoto Roberto Mitsuyoshi 109-P
Hlavacova J 60-O
Hlavačková Kristýna 19-O, 20-O
I
Ilango Kandan 47-P
Infran Jucelei de Oliveira Moura 84-P, 85P, 88-P, 128-P
Intile S 82-P, 23-O
Ipe Ipe M 43-O
Ishikawa Edna Aoba Yassui 76-P
J
Jafari R 40-O
Jiménez Maribel 4-P, 99-P, 130-P, 131-P,
145-P
Joshi Amritanshu 56-O
Juan Laura W 5-P
N° 26 (1) 2015
K
Kahime Kholoud 106-P, 164-P, 165-P,
167-P
Kamhawi Shaden 53-O, 56-O, 115-P
Karakus Mehmet 4-O, 36-P
Kasap Ozge Erisoz 21-O, 39-O, 157-P
Kassahun Aysheshm 19-O
Katakura Ken 15-O
Kato Hirotomo 15-O
Kelly-Hope Louise 8-O, 28-O, 176-P
Khajeian M. 40-O
Khalid Noteila M 66-O
Khoury C 45-O, 68-O
Kindlová Michaela 73-P
Kirstein Oscar 33-O, 34-O, 36-O, 39-P, 41P, 90-P, 162-P
Kobylinski Kevin 7-P
Košťálová Tatiana 73-P
Kowalewski Martín 148-P
Kozhahmetova Madina 9-P
Kratochvílová Tereza 73-P
Kumar Vijay 6-O
Kwakye-Nuako G 100-P
L
Lacerda Helio Márcio Nunes 78-P
Lagarmilla Patricia 122-P, 141-P, 142-P,
150-P
Lal Das Murari 1-O
Lal Dinesh 43-O
Lamballerie X de 44-O
Lana Rosana Silva 79-P
Lara-Silva Fabiana de Oliveira 74-P, 75-P,
79-P
La Russa F 82-P
Lastre Natalia 59-P
Laurenti MD 97-P
Lavitschka Cecilia Oliveira 104-P
Lawson Daniel 62-O
Lazari Thaís Fernanda 171-P
Leão Edna De Freitas 112-P
Léger Nicole 16-O, 24-O
Lehrter Veronique 20-O
Leite Keily Cristiny Azevedo 78-P
Lelli R 82-P
Lemma Wossenseged 33-O, 35-O, 29-P,
34-P
Levay Ana Paula Silva 101-P
Libri Valentina 57-O
Lima Ana Cristina Vianna Mariano da
Rocha 74-P, 75-P, 174-P
Lima Fernanda Pinheiro 13-P
Lima Jan108-P
Lima José Aprígio Nunes 112-P
Lima Magliones Carneiro 15-P, 77-P
Limam S 132-P
Lisi O 45-O
Lizarralde de Grosso MS 163-P
Lopes Josiane Valadão 75-P
Lopes Tamy Elicia da Silva 118-P, 119-P,
146-P, 149-P, 156-P
Lopez Marla 153-P
Lorini Maria Lucia 40-P
Louzir Hechmi 57-O, 58-O
Lozano Alejandra 122-P, 141-P, 142-P,
150-P
Lubbad MY 117-P
Lugo Ligia 153-P, 161-P
Luna R 127-P
LL
Llanes-Acevedo Ivonne Pamela 17-O, 22-O
Llanos A 27-O
M
Macena Ana Lúcia Moreira 15-P, 77-P
Machado Caroline Amaral 22-P, 116-P
Machado Vicente Estevam 71-O, 113-P,
114-P
Machado-Coelho George Luiz Lins 74-P
Maia C 33-P, 57-P, 111-P
Magalhães Mônica de Avelar Figueiredo
Mafra 18-P
Mandal Goutam 115-P
Mandal Srotoswoti 115-P
Marceló Catalina 9-O
Marchi A 68-O
Marchini Fabricio K 59-O, 69-O
Marco JD 158-P
Margonari Carina 13-P
Maroli M 45-O
Mas-Coma Santiago 18-O
Massoudi Nabil 132-P
Mastrángelo AV 11-O
Martín-Martín Inés 4-P, 99-P,
Martín Ohiane 17-O, 22-O, 55-O
Martín-Sánchez J 154-P
Martínez Lily Paola 46-O, 51-O, 91-P
Martínez Mariela F 148-P
Martínez Nidia 133-P
Martins Juliana 79-P
Mateus Nathália Lopes Fontoura 88-P
Mathis Alexander 20-O
Matthew Albert 57-O
Mazloumi Gavagni A 61-P
McCarthy Christina B 3-P
McCoy John Philip 56-O
McDowell Mary Ann 61-O, 63-O,
Meddeb-Garnaoui Amel 57-O, 58-O
Medley Graham 170-P
Meira Antônio de Medeiros 110-P
Melo Alan Lane 80-P
Memish Ziad 28-O
Mendes Adauto Rodrigues 128-P
19
Mendes Ileane Gurgel 119-P
Mendonça Ivete L 10-P
Mendoza J 127-P
Menegatti Jaqueline Aparecida 29-O
Meneghello Brígida Helena da Silva 109-P
Meneses Claudio 115-P, 70-O, 56-O
Menezes Filho Antônio 10-P
Menezes Júlia Alves 13-P
Menz Ingrid 104-P
Mermer Ahmet 37-P
Mhadhbi M 132-P
Michalsky Érika Monteiro 74-P, 75-P, 79-P,
93P
Mierez Mirta Liliana 62-P
Miguelañez Silvia 17-O, 22-O
Miranda Débora Elienai de Oliveira 31-P,
32-P, 152-P
Miró Guadalupe 17-O, 22-O
Miyazaki Rosina Djunko 26-P, 27-P, 28-P,
93-P
Molina Juan 11-O, 5-P
Molina Ricardo 4-P, 99-P, 130-P, 131-P,
145-P
Molla Leticia María 104-P
Monaco Neiva Zandonaide 88-P
Moncaz Aviad 33-O, 34-O, 36-O, 90-P
Mondragon-Shem Karina 28-O, 46-P, 176P
Montesino Ana 51-O, 91-P
Montoya Ana 17-O, 22-O
Moraes Carolina d. 67-O
Morais María Helena Franco 135-P
Morais Mayron Henrique Gomes 13-P
Morales E 70-O
Morilla F 154-P
Moreno Eduardo Stramandinoli 20-P
Moreno H Claudia Ximena 73-O
Mosqueda Luis Armando 62-P
Moura Abilene Cristina de Arruda 32-P,
152-P
Mukhopadhyay Rita 115-P
Műller Ingrid 55-O
Murgas Ingrid 65-O
N
Nakashi Hira L 56-O
Naouar Ikbel 57-O, 58-O
Nascimento Lucimar 172-P
Nascimento Luis Otávio Cordeiro 20-P
Nasser Julio R 67-P, 158-P
Negrão Fábio Juliano 101-P
Neto Grimoaldo Braga Da Rocha 15-P, 77P
Nogueira Fabio dos Santos 104-P
Novo Shênia Patrícia Corrêa 110-P
Nowling RJ 63-O
Nzelu Chukwunonso Onyemaechi 15-O
N° 26 (1) 2015
O
Obara Marcos Takashi 6-P
Oliveira AFJ 151-P
Oliveira Alessandra G 124-P
Oliveira Ana Flávia de Morais 78-P
Oliveira DMS 108-P
OliveiraEverton Falcão 128-P
Oliveira Fabiano 53-O, 70-O
Ölgen M. Kirami 4-O
Oreste F 67-P
Orta Jose 115-P
Ortega Edgar 26-O, 8-P, 63-P
Ortiz Washington Benitez 162-P
Oscherov EB 38-P
Oshiro Elisa Teruya 84-P, 85-P, 88-P, 101-P,
128-P
Osuguku E 27-O
Otranto Domenico 12-O, 152-P
Ouanaimi Fouad 106-P, 164-P, 165-P, 166-P,
167-P
Ovallos Fredy Galvis 70-P, 71-P, 104-P, 173P
Özbel Y 4-O, 39-O, 36-P, 37-P
Özkul A 39-O
P
Pacheco R 27-O, 102-P, 103-P, 126-P, 127P, 147-P
Pandey Shubham 43-O
Paranhos Filho Antonio C 124-P
Pardo Raúl H I 7-O, 9-O, 10-O
Parente Daniela M 10-P
Parrado Rudy 86-P, 87-P
Parras Matías Ariel 23-P, 24-P, 68-P, 69-P,
125-P
ParreiraR. 57-P
Passero LF 97-P
Paternina Luis 46-O
Paternina Margaret 46-O
Paz Gustavo Fontes 139-P
Pech-May Angélica 105-P
Pereira AAS 174-P
Pereira Araujo T 102-P
Pereira-Chioccola 104-P
Pereira Júnior Antonio Marques 98-P, 129-P
Pérez Adriana Alicia 11-O, 43-P
Pérez-Doria Alveiro 46-O, 51-O, 59-P, 91-P
Perez JE 27-O, 103-P, 126-P, 127-P
PessanhaJosé Eduardo Marques 74-P
Pessoa Felipe Arley Costa 98-P, 129-P, 151P
Pessoa Grasielle CD 16-P
Pesson B 154-P
Pflüger Valentn 20-O
Philips-Singh Doris 43-O
Piazza M 23-O
Picado Alberto 1-O
Pinheiro Marco Paulo Gomes 118-P, 119-P,
120-P, 146-P, 149-P, 156-P
Pinheiro Maria Paula de Melo Pereira 120-P
Pinheiro Maria Sueli Barros 92-P, 112-P
Pinto Israel de Souza 116-P
Pinto Mara Cristina 71-O, 113-P, 114-P
Pimenta Paulo PFP 48-O, 52-O
Pitaluga Andrè N 54-O, 69-O
Pita-Pereira D 102-P
Pires Ana Clara Araújo Machado 52-O
Pitti R 23-O
Poché David M 5-O, 9-P
Poche Richard 2-O, 3-O, 9-P
Polyakova L 3-O, 9-P
Ponte Cesar do Santo 110-P
Portillo Nilda 133-P
Portús M 145-P, 154-P
Posada Laura 46-P
Posada-López Laura 73-O, 63-P, 159-P
Pozo Catherine 162-P
Prudhomme Jorian 20-O
Pruzinova K 60-O
Q
Quaresma Patrícia Flávia 25-P, 60-P, 72-P,
95-P
Qualls Whitnay A 155-P
Quintana María Gabriela 11-P, 65-P, 163-P,
168-P
Quispe Anita 27-O, 103-P, 126-P, 147-P
Quispe C 147-P
Quispe D 27-O, 126-P
Quispe-Florez MM 102-P, 103-P
Quispe M 27-O
Quispe-Ricalde MA 103-P, 147-P
R
Rado D 27-O, 102-P, 103-P, 126-P, 127-P,
147-P
Rahola Nil 53-P
Ramalho Miriele 152-P
Ramalho-Ortigao José Marcelo 59-O, 115P
Randrianambinintsoa Fano José 16-O
Rangel Elizabeth F 16-P, 18-P, 19-P, 40-P,
42-P, 48-P, 80-P, 89-P, 102-P, 126-P
Raizer Josué 29-O
Ravel C 145-P
Rea María Josefa Felisa 62-P, 121-P
Ready PD 47-O
Reale S 23-O, 82-P
Rebai-Kammoun Wafa 57-O, 58-O
Rebolar-Téllez Eduardo A 105-P
Remondegui CV 168-P
Rêgo Felipe Dutra , 25-P, 94-P, 95-P, 116-P,
134-P, 138-P, 139-P
Rego Taiana Amâncio da Costa 19-P
20
Reis AS 66-P, 174-P
Reithinger Richard 86-P
Remoli ME 68-O
Rhodes C 70-O
Ribeiro Ana Lucia Maria 1-P, 26-P, 27-P,
28-P, 93-P
Ribeiro Carlos CB 10-P
Ribeiro Leticia Moraes 128-P
Rigg Chystrie 35-P
Ríos Cristina 122-P, 141-P, 142-P, 150-P
Rocha Anárya Teresa de Freitas 123-P
Rodarte Cláudia Cavalcante De Matos 77P
Rodrigues Eduardo Henrique Gomes 12-P
Rodrigues Jorge Senatore Vargas 1-P, 26-P,
27-P, 28-P
Rodrigues Moreno de Souza 129-P
Rodríguez Jorge Luis 51-O
Rodríguez-Jiménez Jorge 91-P
Rocha Douglas de Almeida 6-P, 50-P
Rogers Matthew E 55-O
Rohoušová Iva 73-P
Rojas Agustina 25-O
Rojas de Arias Antonieta 11-P
Romero Luis Roberto 51-O
Romero Selva 122-P, 141-P, 142-P, 150-P
Romero Vanesa 162-P
Romero-Ricardo Luis 59-P, 91-P
Rosa Juan Ramón 23-P, 24-P, 38-P, 68-P,
69-P
Routray Parimita 46-P
Rowland Mark 1-O
Rowland Tobin 7-P
Rowton Edgar 7-P
Ruiz-Lopez Freddy 159-P
Rugani Jerônimo Marteleto Nunes 25-P, 95P, 174-P
S
Sábio Priscila Bassan 51-P, 52-P, 56-P
Sabroza Paulo Chagastelles 135-P
Sadlova Jovana 49-O, 50-O
Saeidi Z 61-P
Salah Afif B. 32-O, 57-O, 58-O
Saldaña Azael 35-P
Salomón Oscar Daniel 11-O, 3-P, 5-P, 11-P,
21-P, 23-P, 24-P, 38-P, 43-P, 44-P, 45-P, 58P, 65-P, 68-P, 69-P, 83-P, 125-P, 148-P,
160-P, 163-P, 175-P
Samantaray Sachidananda 2-O
Sánchez-García Laura 105-P
Sandoval Adolfo Enrique 45-P, 225-P
Sanguinette Cristiani de Castilho 134-P,
136-P
Santamaría Erika 7-O, 9-O, 10-O
Sant'Anna Mauricio R 67-O
Santini María Soledad 11-O, 3-P, 11-P, 21-
N° 26 (1) 2015
P, 45-P, 148-P, 160-P, 175-P
Santos Fabricio K. M. 16-P, 89-P
Santos Luciene Aranha da Silva 112-P
Santos Mirella Ferreira da Cunha 124-P,
128-P
Santos Nathália Queiroz 76-P
Santos TV 108-P
Santos Walter Souza 30-O
Saracho-Bottero MN 163-P
Saraiva Lara 66-P, 136-P, 174-P
Satragno Dinora 122-P, 141-P, 142-P, 150-P
Scholte Ronaldo Guilherme Carvalho 95-P
Sciamarelli Alan 29-O
Sebastien Boyer 16-O
Seblova-Hrobarikova Veronika 49-O, 50-O
Secundino Nágila FC 48-O, 52-O
Sequeira Cirino 141-P, 142-P, 150-P
Ser Önder 4-O
Serra e Meira Paula Cavalcante Lamy
136-P, 137-P
Sevá Anaiá da Paixão 70-P, 71-P
Severini F 45-O
Shahbazi F 40-O
Shaw Jeffrey 41-O
Shenker Moshe 33-O
Shimabukuro Paloma Helena Fernandes
13-O, 20-P, 22-P, 55-P, 94-P, 95-P, 116-P,
171-P
Silva AM 54-O
Silva BJM 108-P
Silva EO 108-P
Silva João Carlos França 79-P
Silva Júlia dos Santos 19-P, 89-P
Silva Maiara Alves 74-P, 75-P
Silva Marcel Miranda de Medeiros 146-P,
149-P, 156-P
Silva Rafaella A 16-P
Silva Simone Melo 123-P
Silveira Fernando Tobias 92-P, 112-P
Silveira FT 97-P, 108-P
Šíma Michal 73-P
Singh Mutum Ingobi 2-O
Singh NS ID 43-O
Soares Daniela Cristina 30-O
Soares Márcia Maria Costa Nunes 109-P
Soares Maria Regiane Araújo 123-P
Soares Rodrigo Martins 70-P, 71-P
Soleimani H 40-O
Soto M 103-P, 127-P
Sousa Alessandra Mara 13-P
Sousa Lindemberg Caranha 16-P, 48-P
Souza Daiana Alovisi 84-P, 85-P, 88-P
Souza Cristian Ferreira 60-P, 72-P
Souza GD 140-P
Souza Mielle Barbosa 60-P
Stanneck Dorothee 12-O
Steinhorst Iris Ingrid 45-P
Stell Frederick 64-P
Sttump Rodolfo German Antonelli Vidal
134-P
Suar Mrutyunjay 46-P
Suau Hamide Aslan 55-O, 115-P
Sumová Petra 73-P
Supparo Eduardo 141-P
Szelag Enrique Alejandro 23-P, 24-P, 68-P,
69-P
T
Tadros Samuel 63-O
Tanure Aline 134-P, 139-P, 139-P
Tartaglino Lilian 3-P
Tavares Mara Garcia 65-O
Tekie Habte 35-O, 36-O, 37-O, 29-P, 34-P,
90-P
Teles Carolina Bioni Garcia 98-P
Telleria Erich Loza 59-O
Tempone Antonio Jorge 54-O, 59-O, 69-O
Thies Sirlei Franck 1-P, 2-P, 27-P, 93-P
Thomaz-Soccol Vanete 11-P
Tinoco B 54-O
Toccas F 27-O, 127-P, 147-P
Tolezano José Eduardo 109-P
Tomokane TV 97-P
Tonelli Gabriel Barbosa 95-P, 134-P, 139-P
Torales Martha 133-P
Torina A 82-P
Torina S 23-O
Tort Cecilia 122-P, 141-P, 142-P, 150-P
Toumi Amine 32-O, 57-O, 58-O
Töz Seray 4-O
Traub-Csekö Yara María 54-O, 59-O, 69-O
Tripet Frederic 66-O
Trueba Gabriel 64-O
U
Uribe Sandra I 8P, 26-O, 63-P, 73-O
Utgés María Eugenia 11-O, 83-P, 125-P,
148-P, 175-P
V
Vaca Franklin 162-P
Vaccalluzzo V 45-O
Valderrama Anayansi 35-P
Vale Mariana Moncassim 40-P
Valencia B 27-O
Valenzuela Jesus 8-O, 28-O, 53-O, 56-O,
70-O, 112-O, 115-P
Valladares B 102-P, 103-P, 126-P, 127-P,
147-P, 176-P
Vargas Jorge Senatore Rodrigues 61-P, 93-P
Vásquez Gisella 64-P
Vega Celeste 11-P
Vela Gabriela 64-O
Vélez Iván Darío 26-O, 8-P, 63-P, 159-P
21
Vélez-Mira Andrès 26-O, 8-P, 63-P, 159-P
Verbel-Vergara Daniel 46-O
Vergara José G 51-O, 91P
Verger Lorenzo 122-P, 141-P, 142-P, 150-P
Vianna Elisa Neves 135-P
Vidal Joaquim 17-P
Viera Ana 122-P, 141-P, 142-P, 150-P
Vilela M 126-P
Villacrés Ernesto 162-P
Villarquide ML 160-P
Vincent Robert 16-O
Vitale Edgardo 122-P, 141-P, 142-P, 150-P
Vitale F 23-O, 82-P
Vivero Rafael J 26-O, 73-O, 8-P, 63-P
Volf Petr 19-O, 20-O, 21-O, 39-O, 49-O,
50-O, 60-O, 73-P
Votýpka Jan 19-O, 21-O, 39-O, 49-O, 60-O
W
Wadsworth Marinha 63-O
Wang Hsiao-Hsuan 5-O
Warburg Alon 19-O, 33-O, 34-O, 35-O, 36O, 37-O, 38-O, 42-O, 49-O, 29-P, 34-P, 39P, 41-P, 73-P, 90-P, 162-P
Ximenes Maria de Fátima Freire de Melo
118-P, 119-P, 120-P, 146-P, 149-P, 156-P
Y
Yadon Zaida E 11-P
Yaghoobi-Ershadi M. Reza 40-O
Yared Solomon 33-O, 38-O
Yazidi Rihab 96-P
Yépez José Yancarlos 25-O
Yilmaz Bahtiyar 4-O
Z
Zacarias Danielle A 10-P
Zanini Volmir 20-P
Zaffora G 23-O
Zapata Sonia 20-O, 64-O
Zenobio Ana Paula Lusardo de Almeida
136-P
Zerba Eduardo N 5-P
Zhioua Elyes 3-O, 72-O, 44-O, 132-P,
155-P
Zoghlami Z 132-P
Zorrilla Victor 64-P
Zwetsch Adriana 48-P
ABSTRACT BOOK
ROUND TABLES
&
ORAL SESSIONS
ISOPS VIII
Puerto Iguazú
Argentina
N° 26 (1) 2015
ID 1-O Vector Control
EVALUATION OF DIFFERENT NETS AGAINST
PHLEBOTOMUS ARGENTIPES, THE VECTOR OF
VISCERAL LEISHMANIASIS IN NEPAL
MULTIPLE FIPRONIL DOSES IN DAIRY CATTLE
FOR VECTOR CONTROL IN BIHAR, INDIA: RESIDUE
STUDY AND EFFICACY DETERMINATION AGAINST
THE SAND FLY PHLEBOTOMUS ARGENTIPES
Albert Picado; Murari Lal Das; Mark Rowland; James Austin; Elisa
De Lazzari
B a rc e l o n a C e n t re f o r I n t e r n a t i o n a l H e a l t h R e s e a rc h ( C R E S I B ) .
albert.picado@cresib.cat
Keywords: Phlebotomus argentipes, insecticide treated nets, mesh
size, cross-over trial
23
Rajesh B Garlapati1, Sachidananda Samantaray2, Mutum Ingobi Singh1,
Trey Barresi1, Dylan Burruss1 and Richard Poche1
1
Genesis Laboratories, Wellington, Colorado, USA. 2Bihar Veterinary College, Patna,
Bihar, India.
rajesh@genesislabs.com
Keywords: Fipronil, cow, residue, sand fly
In the Indian subcontinent, Leishmania donovani, the parasite causing
visceral leishmaniasis (VL) is transmitted by the sand fly vector
Phlebotomus argentipes. Long lasting insecticide treated nets (LN)
have been postulated as alternative or complement to Indoor Residual
Spraying but there are few field studies evaluating the entomological
efficacy of different nets against this vector. We conducted two crossover trials in a VL endemic area in Nepal to compare the barrier effect
of (1) LN with different mesh sizes (156 holes/inch2 vs 625
holes/inch2) and (2) alpha-cypermethrin treated LN and untreated
nets having the same mesh size (156 holes/inch2). Each crossover trial
had two arms consisting of a sequence of two different nets for 8
nights. We used 10 cattle sheds per trial as unit of analysis and a cow
placed under the net as bait. CDC light traps placed inside the nets
were used to evaluate the number of P. argentipes crossing the net
barrier. Negative binomial generalized estimating equation (GEE)
population-averaged models adjusted by night and sequence were
used to estimate the barrier effect of the different nets. The crossover
trials conducted in a rural village in Morang district (South-eastern
Nepal) demonstrated that reducing the size of the holes in treated nets
(625 holes/inch2) increased the barrier effect of LN by 77% (95%
confidence interval (CI): 56% - 88%) compared with treated nets with
larger holes (156 holes/inch2). Treating nets with alpha-cypermethrin
reduced the number of P. argentipes captured inside the nets by 77%
(95% CI: 27% - 93%) compared with untreated nets. The effect of
finer mesh LN on VL prevention needs to be tested in a randomized
controlled trial.
Visceral Leishmaniasis is a protozoan disease prevalent in India. VL
is a severe form of Leishmaniasis which is transmitted by sand flies.
Phlebotomus argentipes is the only available vector for VL
transmission in India. Vector control plays an important role in VL
management. Cows are an important blood meal source for P.
argentipes. A study was initiated to assess sand fly mortality in
response to fipronil oral dosing of dairy cows. Twenty hybrid cows
were selected and four different doses (0.25 mg/Kg body weight, 0.5
mg/Kg body weight, 1.0 mg/Kg body weight and 1.5mg/Kg body
weight) of fipronil were given at 8 week intervals for six months,
totaling three separate applications. Sand fly efficacy and fecundity
were evaluated on Day 1, 4, 7, 14, 21, 28, 35, 42 and 49 after
treatment. Fipronil residues in milk were assessed for the first 14 days
after treatment. Plasma residues were assessed on Day 1, 4, 7, 14, 21
and 28 after treatment during three applications. Milk production,
cow health and environmental conditions were monitored during the
study. Higher doses of fipronil (1.0 mg/Kg body weight and 1.5
mg/Kg body weight) had high residues in milk and longer sand fly
efficacy after treatment. The efficacy of the lowest dose (0.25 mg/kg
body weight) was found up to 14 to 21 days after treatment. Fipronil
did not have a significant effect on sand fly fecundity. Metabolism of
cows played an important role in milk and plasma residues during
three applications. The study results suggest that the use of fipronil as
a feed through substance can be incorporated into Visceral
Leishmaniasis management programs.
N° 26 (1) 2015
1
LABORATORY AND FIELD EVALUATION OF
RODENT BAIT TREATED WITH FIPRONIL FOR FEED
THROUGH AND SYSTEMIC CONTROL OF
PHLEBOTOMUS PAPATASI
M Derbali1, L Polyakova2, A Boujaâma1, D Burruss2, S Cherni1, W
Barhoumi1, I Chelbi1, R Poch2, E Zhioua1
24
Ege University Institute of Health Sciences, Bornova, İzmir, Turkey; 2Ege University
Faculty of Science, Department of Zoology, Bornova, İzmir, Turkey; 3Akdeniz
University Faculty of Science, Department of Biology, Antalya, Turkey; 4Celal Bayar
University Vocational School of Health Sciences, Manisa, Turkey; 5Ministry of Health
Public Health Agency, Antalya, Turkey; 6Ege University Faculty of Medicine,
Department of Parasitology, Bornova, İzmir, Turkey; 7Celal Bayar University Faculty
of Medicine, Department of Parasitology, Manisa, Turkey; 8Ege University Faculty of
L i t e r a t u re , D e p a r t m e n t o f G e o g r a p h y, B o r n o v a , İ z m i r, Tu r k e y.
mehmetk1986@yahoo.com
Keywords: Insecticide, Bednet, Antalya, Turkey
1
Institut Pasteur de Tunis, Laboratory of vector Ecology, 13 Place Pasteur, BP 74, 1002
Tunis, Tunisia. 2Genesis Laboratory Inc., 10122 NE Frontage Road, Wellington, Co
80549, USA.
elyes.zhioua@gmail.com
Keywords: Phlebotomus papatasi, Meriones shawi, zoonotic
cutaneous leishmaniasis, systemic and feed through insecticide
The sand fly Phlebotomus papatasi is the main vector of Leishmania
major, etiologic agent of zoonotic cutaneous leishmaniasis (ZCL),
which is endemic in North Africa, the Middle East, and Asia. In North
Africa, Meriones shawi is one the main reservoir host of L. major.
Phlebotomus papatasi populations are maintained in borrowing
rodents such as M. shawi. Three fipronil-treated rodent baits were
evaluated for systemic and feed through insecticidal activity against P.
papatasi feeding on M. shawi. Through blood feeding bioassays,
mortality rates of females P. papatasi increased with the concentration
of fipronil in the rodent bait varying from 0.001% to 0.005%. In the
laboratory, more than 90.0% of P. papatasi were killed within 48 hours
after blood feeding on the desert's jirds, Meriones shawi, treated up to
29 days prior with a single application of fipronil at a concentration of
0.001%, 0.0025% and 0.005%. Through larval bioassays, mortality
rates of larvae that have fed on faeces of treated bait for M. shawi
increase with the concentrations of fipronil. Faeces of orally-treated
Meriones were significantly toxic to larvae for 5 weeks with a
concentration of 0.005%. In the field, application of treated bait
resulted in 80.0% reduction in the populations of P. papatasi up to 6
weeks after a single application of fipronil at a concentration of
0.005%. This is the first study to demonstrate field efficacy of
fipronil-treated rodent baits for P. papatasi control and the first study
to evaluate this approach in M. shawi, a principal ZCL reservoir host.
These results suggest that fipronil-treated rodent baits can be used to
effectively reduce the populations of P. papatasi associated with M.
shawi in ZCL endemic areas.
N AT U R A L I N F E C T I O N A N D I N S E C T I C I D E
SUSCEPTIBILITY STATUS OF WILD CAUGHT SAND
F L I E S I N R U R A L A R E A S O F A N TA LYA ,
MEDITERRANEAN REGION OF TURKEY
Mehmet Karakuş1; Samiye Demir2; Hüseyin Çetin3; Suha Kenan
Arserim4; Önder Ser5; Seray Töz6; I Cüneyt Balcioğlu7; M Kirami Ölgen8;
Bahtiyar Yilmaz2; Yusuf Özbel6
Sand fly-borne diseases like visceral, cutaneous leishmaniasis and
phlebovirus infections have been seen endemically in Turkey. The
application of insecticides is mainly targeting mosquitoes/flies
control. However, no study was performed about the susceptibility of
sand flies against pyrethroids in Turkey. This study was carried out in
Antalya province located in The Mediterranean Region of Turkey and
we aimed to determine (1) the sand fly fauna, (2) the susceptibility of
wild caught sand flies, mainly collected from rural areas, against
commonly used pyretroids and (3) natural infection of Leishmania.
CDC's bottle bioassay has applied for pyrethroid susceptibility by
using different dosages. The tests were repeated 5 times with 100
specimens per bottle. In addition, WHO's Cone test were performed
using two different bed nets (Olyset Plus® with PBO and Permanet®)
to understand their efficacy. Following the standards of WHO testing
procedures, all specimens were dissected, mounted and identified
using Mediterranean keys. After dissection, the rest of the body of the
specimens was pooled according to the locations and species and
totally 50 pools were generated. DNA extractions were made using
commercial kit and previously described conventional PCR of ITS-1
region was performed. A total of 10 Phlebotomus species were found
in the province with the abundance of P. neglectus (38.82%), P.
alexandri (21.66%) and P. tobbi (20.44%). Totally three different
active ingredients were tested and knock down times were calculated
using Probit Analysis. Knock down times were noted with 10min
intervals, KdT50/KdT100 were noted for determining the effective
dosages. For the lowest dosage of insecticides, KdT100 value were
noted as 36 minutes for permethrin (0.05%), 60 minutes for
cypermethrin (0.05%) and 42 minutes for deltamethrin (0.05%). By
the end of 24-hour period, no sand fly was alive in bottle assay while
46% and 0% of sand flies were alive in cone test-Permanet® and cone
test- Olyset Plus®. Four pools containing Phlebotomus neglectus and
one pool Phlebotomus tobbi were found to be positive for Leishmania
spp. In conclusion, no resistance was determined in bottle assay but
upcoming resistance with prolonging death time and knock down
times were noted. Cone test results have showed that Olyset Plus®
has more knockdown effect than Permanet®. Mediterranean Region
is one of the important tourism and agriculture centers in Turkey.
Related to this the insecticide usage for agriculture and
mosquitoes/flies control is relatively high in this region. Even no
resistance was detected in this study; related to the presence of
important vector species of Leishmania infantum and detecting of
natural infection in the area, the status of susceptibility of sand flies
needs to be monitor regularly in the region.
This study is supported by The Scientific and Technological Research
Council of Turkey (Project No: 112T270).
N° 26 (1) 2015
SAND FLY CONTROL IN BIHAR, INDIA: MODELING
ALTERNATIVE CONTROL STRATEGIES USING
SYSTEMIC INSECTICIDES ORALLY ADMINISTERED
TO LIVESTOCK
DEVELOPMENT OF RESISTANCE AGAINST DDT TO
PHLEBOTOMUS ARGENTIPES (DIPTERA:
PSYCHODIDAE) AND SEARCH FOR ALTERNATIVE
INSECTICIDE
David M Poché, William E Grant, Hsiao-Hsuan Wang
Diwakar Singh Dinesh, Faizan Hassan, Vijay Kumar and Pradeep Das
Texas A&M University, Genesis Laboratories, Inc.
Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical
Research), Agamkuan, Patna-800007, India.
davidp@genesislabs.com
25
drdsdinesh@yahoo.com
Keywords: Bihar, fipronil, Phlebotomus argentipes, simulation
model.
Approximately 60 percent of visceral leishmaniasis (VL) cases occur
in Northern India and bordering regions of Nepal and Bangladesh,
where the disease is spread by the sand fly species Phlebotomus
argentipes. Bihar is the most poverty-stricken state in India and the
majority of VL cases reported in the country occur in this region.
DNA extraction suggests that cattle, domestic buffalo, goats, and
humans serve as the primary hosts for P. argentipes in villages in
Bihar. Recent research in Bihar has indicated that the systemic
insecticide fipronil, orally administered to cattle, can kill 100% of
adult P. argentipes feeding on insecticide-treated cattle and also kill
100% of larvae feeding on feces from insecticide-treated cattle. In
addition, fipronil is a lipophilic compound and has the unique ability
to be efficacious for several weeks. These results have suggested that
fipronil can serve as a means for controlling sand fly populations and
potentially reduce cases of VL in the region. To evaluate the potential
efficacy of large scale fipronil treatment, a quantitative model was
developed, representing the various sand fly life stages (egg, larva,
pupa, and adult) in a typical rural village in Bihar. Seasonal variations
in temperature, precipitation and relative humidity were incorporated
into the model to better represent P. argentipes seasonality in Bihar
within the system of interest. A variety of control strategies were
simulated in which we varied the season and frequency of livestock
treatment. Simulation results suggest that the potential to reduce the
sand fly population in Bihar will depend on coordination of the timing
of treatments with particular phases of the P. argentipes life cycle as
they relate to sand fly seasonality in Bihar. The simulations also
suggest that control strategies should be administered over a multiyear period to achieve preferred results.
Keywords: Sand fly DDT resistant
Phlebotomus argentipes (Diptera: Psychodidae) Annandale and
Brunetti, the vector of Visceral Leishmaniasis in Bihar, India is
developing resistance against the insecticide of choice, i.e. DDT. The
measurement of tolerance/resistance was based on tube-bioassay test
with 4% DDT impregnated paper to the laboratory bred newly
emerged unfed and blood fed along with wild caught fed female P.
argentipes. Variable results were found showing 40-60% resistant
population in all tested P. argentipes. This was verified and compared
with changes in Glutathion S- transferase (GST) enzyme (major
detoxifying enzyme for xenobiotics) at protein level. The diagnostic
band of 42 kD was found highly expressed among the resistant group
by the 12% SDS- PAGE in the lab colonized and wild caught P.
argentipes that was also confirmed as GST through Western Blot.
Other enzymes like Acetylcholinesterase, Esterase and Mono
oxygenase enzymes are under process of evaluation. Further,
alternative insecticide like Deltamethrin was tried and found very
effective in killing P. argentipes in tube bioassay test conducted with
0.05% WHO impregnated paper corresponding with the result of
indoor residual spray 20mg/m2. This study will lead in finding actual
number of resistant population along with the use of an alternative
effective insecticide to control transmission of the disease.
N° 26 (1) 2015
T O X I C A N D B E H AV I O U R A L E F F E C T S I N
EXPERIMENTAL HUTS OF LONG LASTING
INSECTICIDE TREATED NETS AGAINST LUTZOMYIA
LONGIFLOCOSA IN THE SUB-ANDEAN RURAL ZONE
OF COLOMBIA
A CONTROL STRATEGY FOR OLD WORLD
CUTANEOUS LEISHMANIASIS OUTBREAKS
Erika Santamaría, Olga Lucía Cabrera, Raul H Pardo
Grupo de Entomología, Instituto Nacional de Salud, Bogotá. Grupo de
Entomología y Enfermedades Transmitidas por Vectores, Universidad de La
Salle, Bogotá.
esantamaria@ins.gov.co
26
Waleed Al Salem1,2*, Louise Kelly-Hope1,3, Maha Abdeladhim4, Salah
Balgonaeem2, Mohammed Al Zahrani2, Jesús Valenzuela4, Álvaro
Acosta-Serrano1,5
1
Parasitology Department, LSTM, England, 2Saudi Ministry of Health, Riyadh, Saudi
Arabia, 3Neglected Tropical Disease Centre, LSTM, England, 4Vector Molecular
Biology Section, National Institute of Allergy and Infectious Diseases, National
Institutes of Health, Maryland, USA, 5Vector Biology Department, LSTM, England.
waleed-alsalem@hotmail.com
Keywords: Lutzomyia longiflocosa, long lasting insecticide treated
nets, efficacy, experimental hut
Keywords: Old world cutaneous leishmaniasis, Saudi Arabia, Al
Ahsa
The aim of the study was to evaluate the toxic and behavioral effects of
long-lasting insecticide-treated nets (LLNs) against Lutzomyia
longiflocosa in the sub-Andean rural zone of Colombia in the first
experimental hut designed for sand flies. Three bednets were tested:
Permanet 2.0 (a.i.: deltamethrin, 55 mg/m2), bednet treated with Icon
Maxx (a.i.: lambda-cyhalothrin, 60 mg/m2), and control (untreated
bednet). Several holes were made on the bednet to simulate the
conditions of a torn net. In each hut, two volunteers were located
inside the bednet. The study design was a 3 x 3 multiple Latin square.
The recorded variables were: number of sand flies inside the hut and
within the exit traps, human landing rate, and mortality. The detected
effects of the LLNs on L. longiflocosa were: 1) Inhibition from
entering the net: the percentages of females found inside the nets
treated with Icon Maxx (13.9%) and Permanet (2.0, 44.8%) were
significantly lower compared to the control (77.1%). The Icon Maxx
showed the greatest difference in percentage of females inside the net
compared with the control, 62.3% (59.0 - 69.4), twice the difference in
percentage of females found inside the Permanet, 32.3% (27.7- 36.9).
2) Landing inhibition: The mean human landing rate was 10 times [4.9
females/night/2 persons (f/n/2 p)] and three times (15.1 f/n/2 p) lower
in the Icon Maxx and Permanet (2.0) respectively, compared with the
control, 49.5 f/n/2 p. The protective effect of the Icon Maxx (90.1%)
was higher compared to the Permanet (69.5%. 3) Repellency: the
mean exit rate of L. longiflocosa was significantly higher in the Icon
Maxx (14.1 f/n) compared to the control (5.1 f/n). Furthermore, the
percentage of resting females on the walls of the hut with the LLNs
(using as denominator the total number of females outside the net) was
significantly lower with the Icon Maxx (23.3%) and Permanet
(30.5%) compared to the control (72.3%. 4). Mortality: Mortality at 0
h post-test with the Icon Maxx was almost twice (52.5%) that recorded
with the Permanet (28.6%). Nevertheless, mortality at 24 h was
similar for both LLNs (78.8% and 82.6%) respectively, without
significant differences. The two LLNs showed strong effects on L.
longiflocosa behavior and mortality. However, the Icon Maxx seems
better because of its quicker lethal effect, higher protective effect,
inhibition from entering the net and repellency.
Old World cutaneous leishmaniasis (CL) is one of the most prevalent
vector-borne diseases in the East Mediterranean Region, including
the Kingdom of Saudi Arabia (KSA). A combination of factors is
responsible for the sustained spread of CL throughout this region,
including lack of health education, socio-economic factors, civil
unrest, and human migration. Furthermore, uncontrolled
urbanization and lack of governmental sector integration (i.e.
Ministries of Health and Agriculture, and Municipalities) impede the
establishment of a coordinated disease control strategy. Our case
study focused on a new construction site in a remote area of Al Ahsa
governorate (Eastern KSA), which had not been previously assessed
for CL by the local leishmaniasis control team. This region is known
for being exclusively endemic for zoonotic CL (1, 2). Prior to the start
of the leishmaniasis transmission season (~April 2012), 150 (nonlocal) construction laborers, with no previous history of CL, arrived
at the construction site. In January 2013, ~60% of the construction
laborers were reported to have acquired CL, and further PCR-RFLP
analysis confirmed they were infected with Leishmania major (2).
Furthermore, the same cohort of patients presented higher serum
levels of anti-SP32 antibodies (a known Phlebotomus papatasi saliva
marker (3)) compared to infected local residents living in neighboring
CL-controlled areas. Over the course of 2013 (March-November),
an integrated disease control strategy, conducted by several
governmental sectors, which consisted of rodent (mechanical) and
vector control, was applied in and around the construction site.
Mechanical control involved the removal of Hammada elegans
(rodent food plant) and destruction of rodent burrows within a radius
of 500 m around the construction area. In addition, vector control was
employed by spraying deltamethrin into rodent burrows and wall
cracks. Since the implementation of this integrated control strategy,
no CL cases have been reported at the construction area up until April
2014.This case study suggests that (i) implementation of an
integrated vector and rodent control program is sufficient to
overcome outbreaks of zoonotic Old World CL and, (ii) assessing
potential disease transmission areas prior to the establishment of new
development, is essential to prevent CL outbreaks.
N° 26 (1) 2015
27
LONG LASTING INSECTICIDE TREATED NETS
DECREASE SANDFLY ABUNDANCE AND BLOOD
FEEDING SUCCESS INDOORS IN THE SUB-ANDEAN
REGION OF COLOMBIA: RESULTS OF A HAMLET
VECTOR CONTROL TRIAL AFTER ONE YEAR
FOLLOW UP
FIRST EXPERIMENTAL HUT TO STUDY THE
EFFECT OF INSECTICIDE CONTROL MEASURES
AGAINST SANDFLIES (DIPTERA: PSYCHODIDAE)
Olga Lucía Cabrera, Erika Santamaría, Raúl H Pardo
Instituto Nacional de Salud
Raúl H Pardo, Olga Cabrera, Catalina Marceló, Erika Santamaría
olgalucabrera@yahoo.com
Universidad de La Salle.
rpardo@unisalle.edu.co
Keywords: experimental hut, sandflies, control
Keywords: control, Lutzomyia longiflocosa, bednets
Experimental huts have been used to evaluate the effects of
insecticide control measures against mosquitoes indoors mainly for
malaria vectors. Such huts have not been used for sand flies. The aim
of this study was to build an experimental hut in which to evaluate
insecticide control measures against sand flies. The study was carried
out in the sub-Andean coffee area of Colombia in an endemic area for
cutaneous leishmaniasis (CL) where Lutzomyia longiflocosa is
abundant. The project was conducted in three parts: 1) Avenues of
entrance and exit. To identify the main avenues of entrance and exit
into a rural house, two observational studies on the entering and
exiting behavior of sand flies were conducted. The resulting data
were used to determine trap placement in the experimental hut. For
entrance sites the numbers of sand flies attempting to enter the house
through large openings (sticky trap catches) and narrow slits (cage
trap catches) were compared; while for exit sites the numbers of
blood-fed females previously released within the house, trying to
escape through the openings described above were compared. The
results from both experiments indicate that L. longiflocosa enter
(95% of 313 females) and exit (96% of 177 females) the house mainly
by large openings between the ceiling and walls; 2) Trap selection. To
select the type of traps to use in the hut, laboratory tests were
conducted with Lutzomyia longipalpis in a wind tunnel. Two traps,
modified for collecting sand flies were evaluated based on their
ability to allow the sand flies to enter and prevent them from exiting
the trap. The results show that the horizontal slit trap was the best
trap, allowing 84.1% of females to enter and preventing 87.3% of
those that entered from escaping; 3) Hut construction. Based on the
afore mentioned results, three experimental huts were built in the
field. Each hut was a 2.5x2.5x2.3 m room made of bricks with metal
roof, ceiling and two large openings in each wall, between the ceiling
and the wall, at which to locate the horizontal slit trap system. Special
care was taken to ensure to block all other possible escape sites; and
4) Experimental hut test. Hermeticism was evaluated by releasing
300 females inside each hut. After eight hours 70% of the released
females were recaptured. Observations of human landing catches of
L. longiflocosa inside the experimental huts compared with a control
house (hut without traps) showed that L. longiflocosa entered the
experimental huts in lower numbers than the control house. By
adding baffle traps to the entrances of the experimental huts it was
shown that sand flies entered the experimental hut and the control in
similar densities.
The aim of the study was to evaluate the entomological effect of long
lasting insecticide treated bednet (LLNs) against Lutzomyia
longiflocosa, a presumed main vector of cutaneous leishmaniasis in
Colombia. The study was carried out in the mountainous coffee area of
Huila department. The evaluated variables were: sandfly female
abundance, blood feeding rate, feeding success (more than 75% of
abdomen filled with blood) and mortality. The study was carried out at
the hamlet level using a randomized block design, were 7 triplets of
hamlets were formed based on pre-intervention sandfly abundance.
Within each triplet three treatments were assigned: (i) Permanet
(industrially treated), (ii) Icon Maxx (home treated) and (iii) control
(untreated bednet). The study had three phases: 1) baseline study. 21
selected hamlets were visited to collect pre-intervention data,
including sandfly abundance. Sandflies were collected by CDC light
traps in at least 50% of houses and forest, 2) Intervention. Around 10%
of the highest sandflies abundance houses were selected as
representative for each hamlet, all beds in each hamlet were covered
with a bednet, and 3) pos-intervention. Sandflies abundance was
monitored at 1, 6 and 12 month time-points pos-intervention. Preintervention L. longiflocosa indoors mean abundance was similar by
treatment. Total abundance pos-intervention decrease significantly
from 14.7 f/trap/night in the control to 5.0 f/trap/night for Icon Maxx
and 4.3 f/trap/night for Permanet. It means a reduction of around 70%
in sandfly abundance in each of the LLNs. Comparison for each of the
three month time-points showed the same significant reduction.
Percentages of blood feeding success for the pre-intervention were
significantly different between treatments, so comparisons preintervention vs. pos-intervention were made for each treatment.
Percentage of blood feeding success decreased significantly in the
LLNs with a reduction of 41.5% for Icon Maxx and 28.1% for
Permanet, while the control had a significant increase of 84%.
Sandflies abundance in forest was apparently reduced in the treatment
with Permanet but it increased in the treatment with Icon maxx. The
two LLNs reduce sandfly abundance indoors and blood feeding
success in L. longiflocosa. Icon Maxx could be a useful alternative to
become untreated bednets and treated bednets with reduced
insecticide power to LLNs. Mass killing effect of LLNs against
sandflies could not be discarded.
N° 26 (1) 2015
EFFICACY OF IMPREGNATED BEDNETS AND
SPECIES COMPOSITION IN
PREVENTION OF CANINE LEISHMANIOSIS IN
NATURALLY EXPOSED DOGS USING A 10%
IMIDACLOPRID/ 4.5% FLUMETHRIN COLLAR
(SERESTO®)
EXPERIMENTAL HENHOUSES
Manteca Acosta Ma, b; Molina Ja; Utgés M Ea, b; Mastrangelo AVb, c; Pérez
AAd; Santini M S.b; Salomón ODa, c
REDILA a Instituto Nacional de Medicina Tropical (INMeT); b Centro Nacional de
Diagnóstico e Investigación en Endemo-epidemias (CeNDIE); c Consejo Nacional de
Investigaciones Científicas y Técnicas (CONICET); d Departamento de Ecología,
Genética y Evolución. FCEyN (UBA).
28
Domenico Otranto1; Filipe Dantas-Torres1,2; Emanuele Brianti3;
Fabrizio Solari Basano4; Dorothee Stanneck5; Katrin Deuster5
1
University of Bari, Italy; 2Oswaldo Cruz Foundation, Brazil; 3University of Messina,
Italy; 4Acroblu s.r.l., Italy; 5Bayer Animal Health GmbH, Germany.
katrin.deuster@bayer.com
mariana.manteca@gmail.com
Keywords: impregnated fabrics, vectorial control, sandfly
The use of insecticide-impregnated fabrics as preventive tools has
become a relevant topic of research in the epidemiology and control of
vector-borne diseases because of their ability to reduce vector-human
contact and acceptability by end users at the local level in communitybased vector management programs. In this study we used noncommercial curtains with the intent of finding a participative control
strategy where users can make and use the curtains in their own
community. The project was developed in a rural area (Misiones,
northeastern Argentina) where epidemic outbreaks of cutaneous
leishmaniasis (CL) occurred in 2004-2005 vectored by Nyssomyia
whitmani. The objective was to evaluate changes in sand fly
abundance, particularly of Ny. whitmani, in the presence of 10%
permethrine-impregnated mosquito-nets under semi-field conditions.
Three experimental henhouses were installed on the border of the
forest, each with three chickens. The treatments (impregnated curtain,
IC) and controls (non-impregnated curtain, NIC; without curtain,
WC) were assigned randomly. To simulate incomplete coverage of
human dwellings, curtains were hung so as to cover only two sides of
the henhouses. During two consecutive nights, a light trap was set
inside each henhouse and in a site without chickens (WCH) from 5 pm
to 9 am to measure sand fly abundance.). Trials were repeated monthly
from February 2012 to January 2013 except for the period April-June
when sand fly populations were extremely low. All sand flies were
counted to calculate total abundance and a 10% sample of each
combination of month and treatment was used to determine species
relative abundance. A total of 38,277 sand flies was collected
representing eight species )as follows: Ny. whitmani (89.9%),
Migonemyia migonei (5%), Pintomyia pessoai (2.7%), Bruptomyia
sp. (1.7%), Lutzomyia longipalpis (0.4%); Evandromyia corteleziisallesi, Psathyromyia shannoni and Lutzomyia monticola (<0.1%).
Nyssomyia whitmani was the most abundant species regardless of
treatment (>85%). The trap index (average number of sand flies
collected/trap/night) per treatment was IC (63.6, SEM 21.5) and NIC
(160.2, SEM 62.3), both lower than WC (454.2, SEM 160.1)
(F3,14=5.6, p=0.005). We did not find differences between IC and
NIC, nor between IC and WCH (59.3, SEM 19.2). These results show
significant efficacy both in the use of the impregnated and nonimpregnated curtains as preventive barriers to reduce human-sand fly
contact.
Keywords: Prevention, canine leishmaniosis, imidacloprid,
flumethrin
Field evaluations were conducted to investigate the protective
efficacy of a collar containing 10% imidacloprid/ 4.5% flumethrin
(Seresto®) for the prevention of Leishmania infantum infection in
dogs at different time points (2011-2013). Three animal shelters, two
in Sicily and one in Apulia, located in hyper-endemic areas for L.
infantum with confirmed presence of competent vectors were
selected and 340 dogs testing negative by serology (IFAT) and PCR
on skin and bone marrow were enrolled. Dogs were randomly
allocated to collar-treatment (165) or untreated control (175) group.
The collars were applied at the beginning of the transmission seasons
according to label instructions and only replaced if lost or required for
body weight changes. Blood, bone marrow and skin samples were
collected throughout the study and at the final follow-up. Serological,
cytological and molecular tests were performed to detect the presence
of L. infantum in the different tissues. In Apulia, dogs were ≤6 months
of age at enrolment and of various breeds and had not been exposed to
sand flies previously. None of the collar-treated dogs tested positive
for L. infantum throughout follow-up, whereas 21 of 58 untreated
control dogs tested positive throughout the follow-up with five
animals also displaying clinical signs at the final time point. The
cumulative incidence density rate (IDR) observed in untreated
animals was 46.2%. In Sicily, dogs were 2 months to 8 years old at
enrollment and of various breeds. Three of 102 dogs in the collartreatment group and 47 of 117 untreated control dogs tested positive
throughout the follow-up. The cumulative IDR observed in untreated
animals was 60.7%. The collar (10% imidacloprid/ 4.5% flumethrin)
was demonstrated to be safe and highly effective (93.4–100%, based
on IDRs) in preventing L. infantum infection throughout the season in
heterogeneous dog populations under varying environmental
conditions. The regular use of treated collars during the sand fly
season thus represents a reliable and sustainable strategy for the
control of leishmaniasis in dogs living in or travelling to endemic
areas.
N° 26 (1) 2015
29
ID 13-O Taxonomy & Phylogeny
A NEW PROPOSAL FOR THE TAXONOMIC
IDENTIFICATION OF PHLEBOTOMINE SAND FLIES
(DIPTERA: PHLEBOTOMINAE) IN BRAZIL
SEX PHEROMONES AS A PRE-MATING SPECIES
ISOLATION BARRIER IN BRAZIL
Andrey J de Andrade , Paloma H F Shimabukuro , Eunice A B Galati
V Carter1, DP Bray1, G Borges-Alves1, E Dilger2, O Courtenay2, R Brazil3
and JGC Hamilton1
1
1
1, 3
2
1
Departamento de Epidemiologia, Faculdade de Saúde Pública, Universidade de São
Paulo, Av. Dr. Arnaldo, 515, Pinheiros, São Paulo, SP, 01246-904, Brazil, 2Centro de
Referência Nacional e Internacional para Flebotomíneos, Centro de Pesquisas René
Rachou, Instituto Oswaldo Cruz, Av. Augusto de Lima, 1715, Barro Preto, Belo
Horizonte, MG, 30190-002, Brazil, 3Laboratório de Parasitologia Médica e Biologia
de Vetores, Área de Patologia, Faculdade de Medicina, Universidade de Brasília,
Campus Universitário Darcy Ribeiro, Asa Norte, 70910-900, Distrito Federal, Brazil.
Centre for Applied Entomology and Parasitology, School of Life Sciences, Huxley
Building, Keele University, Staffordshire, ST5 5BG, United Kingdom; 2School of Life
Sciences, University of Warwick, Coventry, CV4 7AL, United Kingdom; 3Fundaçao
Oswaldo Cruz, Instituto Oswaldo Cruz, Lab Doenças Parasitarias, Rio de Janeiro,
Brazil.
j.g.c.hamilton@keele.ac.uk
bioandrey@gmail.com
Keywords: pheromone, species complex, reproductive barrier
Keywords: identification key, taxonomy, type-species, Brazilian
sand fly species
Currently, there are 269 valid phlebotomine sand flies species
recorded in Brazil, which represents the highest diversity of sand flies
when compared to other South American countries. Most species
(approximately 110) were described between 1930 and 1950 by
Brazilian researchers following different classifications and
taxonomic proposals. Systematics of sand flies has been debated
during the last decades, new classifications have been proposed, and
relationships between these species are being studied based on
morphological and molecular characters. Dichotomous identification
keys are the first step towards specific identification and for Brazilian
sand flies the last identification key was published in 2003. The aim of
this study is to present an updated new key to indentify sand fly species
recorded in Brazil. To produce the key type-species deposited in nine
Brazilian institutions were examined. One hundred and sixty nine
species including males and females were found. Observations were
made regarding the preservation status of the slides - whether the
specimens were still visible and labels were adequately presented; the
presence of microscopic slides according to the original description;
and specimens numbers and typology. All type-species were
photographed to build an image dataset. When types were not found,
additional specimens of particular collections were examined.
Drawings of the genitalia were made using a camera lucida and
covered by ink pen. A list of geographical distribution for Brazilian
states, including taxonomic synonyms, and a taxonomic catalog were
compiled. The key is divided by gender and the species are grouped by
morphological characters rather than by phylogenetic relationships. A
total of 251 and 179 dichotomies for 258 males and 235 females,
respectively, are presented in the key. Researchers of public
institutions and health services will be invited to test the taxonomic
key, so it can be validated. An interactive version of this key will be
produced using the DELTA (Description Language for Taxonomy)
software package. It will include all data compiled by us. The DELTA
system allows a quick update as new species will be recorded and
described in Brazil. Financial support: FAPESP.
Members of the Lutzomyia longipalpis species complex in Brazil can
be distinguished through morphological, physiological and
biochemical markers. Determining the relative importance of the
different members of the L. longipalpis species complex in
transmitting visceral leishmaniasis is important for understanding the
epidemiology and control of the disease. It has been proposed that the
species complex is under an incipient speciation process, and one
factor influencing speciation is mate-finding through male sex
pheromones. Male L. longipalpis may be one of four possible sex
pheromone chemotypes: (S)-9-methylgermacrene-B (9MGB), 3methyl-alpha-himachalene, cembrene 1 or cembrene 2. Previous
studies of crosses between different chemotypes carried out in the
laboratory showed differing degrees of reproductive isolation. Thus
the sex pheromones of L. longipalpis could affect pre-mating
behaviour and prevent random mating and gene flow between
individuals. However, field evidence for pheromones acting as premating barriers is lacking. We investigated the cross-attraction of L.
longipalpis, from two different pheromone chemotypes in Brazil
(9MGB from Araçatuba, Sao Paulo State and cembrene 1 from
Marajo, Para State). Using experimental chicken boxes and
pheromone lures containing either pheromone or blank controls,
CDC light traps were used to capture sand flies attracted to the site.
Both male and female sand flies were significantly more attracted to
their conspecific pheromone chemotype than CDC traps containing
no pheromone. Sand flies were not significantly attracted to sex
pheromone of the alternative chemotype. These results support the
contention that male sand fly sex pheromones act as a pre-mating
isolation barrier in the field, reduce non-productive mating
encounters and therefore may be one of the strongest influences on
speciation in the L. longipalpis complex.
N° 26 (1) 2015
IDENTIFICATION OF PERUVIAN SAND FLY SPECIES
(DIPTERA: PSYCHODIDAE) THROUGH DNA
BARCODES
ORIGINS AND AFFINITIES OF MALAGASY
PHLEBOTOMINE SANDFLIES
1
2,3
Chukwunonso Onyemaechi Nzelu , Abraham G Cáceres , Ken
Katakura1, Yoshihisa Hashiguchi4,5,6 and Hirotomo Kato1
1
Laboratory of Parasitology, Department of Disease Control, Graduate School of
Veterinary Medicine, Hokkaido University, Sapporo, Japan; 2Departamento
Académico de Microbiología Médica, Facultad de Medicina Humana, Universidad
Nacional Mayor de San Marcos, Peru; 3Laboratorio de Entomología, Instituto
Nacional de Salud, Peru; 4Department of Parasitology, Kochi Medical School, Kochi
University, Japan; 5Prometeo, Secretaria Nacional de Educacion Superior, Ciencia,
Tecnologia e Innovacion (SENESCYT), Ecuador; 6Centro de Biomedicina, Universidad
Central del Ecuador, Ecuador.
30
Randrianambinintsoa Fano José1; Léger Nicole2; Boyer Sébastien1;
Robert Vincent3; Depaquit. Jérôme2
1
Unité d'Entomologie Médicale, Institut Pasteur de Madagascar, Antananarivo,
Madagascar. 2Université de Reims Champagne-Ardenne, ANSES, EA4688-USC «
VECPAR », Reims, France. 3MIVEGEC, UMR, Montpellier, France.
fanojose@pasteur.mg
Keywords: Phlebotominae, Biogeography, Madagascar,
Systematics
nonsy06@yahoo.com
Keywords: molecular taxonomy, DNA barcode, sand flies, vectors.
Understanding the ecological and medical importance of sand flies
relies on accurate identification of species. Currently, the
identification of phlebotomine sand flies is based on morphological
characteristics. However, this traditional taxonomy is laborious, time
consuming and often complicated by cryptic species complexes.
Molecular approaches have become increasingly employed and
promise to be useful taxonomic markers for rapid and reliable species
identification. The use of DNA barcodes has been proposed recently
as a tool for identification of the species in different animal taxa. In this
study, we explored the efficiency of DNA barcode approach, based on
cytochrome c oxidase subunit I (COI) sequences, for species
identification of Peruvian sand flies, which included major vector
species. A total of 89 sand fly specimens belonging to sixteen
morphological species and two genera (Lutzomyia and Warileya) were
analyzed. We were able to recover and align the target COI fragment
from all sand fly species examined. Neighbor-joining (NJ) analysis of
the DNA barcode sequences show that each sand fly species formed
barcode clusters with tight cohesion that were clearly distinct from
those of allied species and were largely congruent with the
conventional taxonomy. Thus, the results validated the usefulness of
DNA barcoding systems in species recognition in sand flies from Peru.
Until now, the Phlebotomine sand fly fauna (Diptera, Psychodidae,
Phlebotominae) of Madagascar remained largely unexplored and
unknown according to the lack of autochthonous transmission of
leishmaniasis in this country. To date, three phlebotomine sandflies
genera including three endemic subgenera have been recorded from
Madagascar Island.: fourteen species are already described. Since
2000, field trap phlebotomine sandflies were studied using both
morphology and molecular biology (rDNA and mtDNA).One
objective is to understand the origins and the affinities of this fauna
group that was never studied due to the lack of precise knowledge on
the origins of different lineages of these sandflies. At the light of the
results obtained including molecular characterization and
phylogenies, the authors discuss the endemism concerning
Phlebotomine sandflies from Madagascar, the relationships of
Malagasy phlebotomine sand flies with species from Africa, America,
Oceania and Asia, and propose an hypotheses explaining the
settlement of this island by Phlebotomine sandflies, including an old
one related to the isolation of Madagascar from the Gondwana
following a generalized track compatible for the Malagasy
Phlebotomus and probably the Vattieromyia. The morphological
affinities of the later subgenus with two Australian species S. pugifera
et S. standfasti or with the American Pressatia is discussed. The
individualization of the Trouilletomyia and Grassomyia should be
younger and underlined several routes of settlement of Madagascar.
N° 26 (1) 2015
31
DNA BARCODING OF MEDITERRANEAN
PHLEBOTOMINE SANDFLIES BASED ON THE
SEQUENCE ANALYSIS OF THE MITOCHONDRIAL
CYTOCHROME OXIDASE I GENE
A MOLECULAR PHYLOGENY OF THE
PHLEBOTOMINAE
Jérôme Depaquit, Maria Dolores Bargues, Santiago Mas-Coma
Israel Cruz, Rosa Gálvez, Oihane Martín, Ivonne Pamela LlanesAcevedo, Carolina Arcones, Rocío Checa, Ana Montoya, Carmen
Chicharro, Silvia Miguelañez, Guadalupe Miró
WHO Collaborating Center for Leishmaniasis, Servicio de Parasitología, Centro
Nacional de Microbiología, Instituto de Salud Carlos III. Ctra. Majadahonda-Pozuelo
Km2, 28220 Majadahonda-Madrid, SPAIN Departamento de Sanidad Animal,
Facultad de Veterinaria, Universidad Complutense de Madrid. Avda. Puerta de Hierro
s/n, 28040 Madrid, SPAIN.
cruzi@isciii.es
Keywords: Sandflies, Mediterranean, DNA-barcoding, COI
Genomic approaches to taxonomy rely on the variability of DNA
sequences to identify organisms. In this sense, these sequences can be
considered as genetic barcodes. Mitochondrial genes seem to be the
most appropriate candidates for DNA barcoding, among them
cytochrome c oxidase I (COI) possess a greater range of phylogenetic
signal than any other mitochondrial gene and it has been suggested as
the core of a global bioidentification system for animals. DNA
barcoding is increasingly used in phlebotomine sandflies taxonomy,
using either COI or cytochrome-B (cytb) genes. We aimed to assess
the use of COI-based DNA barcoding using sandflies collected from
different regions in Spain, for which we sequenced a 658 bp fragment
of the COI gene. We completed this data set with DNA sequences
available in the GenBank for species prevalent in the Mediterranean
region. In total 156 DNA sequences from 4 different phlebotomine
sandflies species (Phlebotomus ariasi, P.papatasi, P. perniciosus, and
Sergentomyia minuta) from Spain were generated for this work and
added to 129 DNA sequences from 14 different phlebotomine species
(P. alexandri, P. ariasi, P. bergeroti, P. chadlii, P. longicuspis, P.
papatasi, P. perfiliewi, P. perniciosus, P. riouxi, P. sergenti, S.
antenata, S. clydei, S. fallax, S. minuta), from 6 different countries
(Algeria, India, Italy, Malta, Portugal, Tunisia) obtained from the
GenBank. Sequences were aligned and trimmed to get a final
alignment of 620 nt. Intraspecific variability was observed in all
species but P. bergeroti (a single DNA sequence was available), and
haplotypes were defined on the basis of single nucleotide
polymorphisms. Distances were estimated with the Jukes Cantor
model to confirm the data set was amenable for Neighbor-Joining
analysis. With the refined alignment a Neighbor-Joining Tree was
obtained using Kimura 2-parameters model of evolution and 2000
bootstrap replicates. All species included in the tree were supported by
bootstrap values of 98% or higher, indicating that this approach is a
solid candidate for DNA barcoding of phlebotomine sandflies. This
work represents a strong contribution to the collection of COI DNA
sequences available for phlebotomine sandflies from Spain. We also
discuss different approaches to DNA barcoding using other targets,
such as cytb, as well as other analysis methods and models of
evolution. Acknowledgement: COLCIENCIAS/COLFUTURO
2012, Instituto de Salud Carlos III MPY-1248/12.
Université de Reims Champagne-Ardenne, ANSES, EA4688 – USC.
jerome.depaquit@univ-reims.fr
Keywords: Phylogeny, ribosomal DNA, paleobiogeography.
Phlebotomine sandflies belong to the subfamily Phlebotominae
(Diptera, Psychodidae). The systematics of this group is the
inheritance of many typological systematics carried out in the Old
World (Abonnenc 1972, Lewis 1982, Artemiev and Neronov 1984,
Seccombe, Ready et al. 1993) and in the New World (Young and
Duncan 1994 ). However, a few phylogenetical and phenetical studies
at the subfamily scale have been carried out (Galati 1995, Rispail and
Léger 1998, Rispail and Léger 1998, Galati 2010) based on
morphological characters.We propose a molecular phylogeny of the
Phlebotominae based on specimens from Africa, Europe, Asia,
Oceania and the Americas. Our sampling includes the following
genera:
Warileya, Spelaeophlebotomus, Idiophlebotomus and
Chinius regarding the Hertigiini; Phlebotomus (subgenera
Phlebotomus, Paraphlebotomus, Synphlebotomus, Larroussius,
Adlerius, Transphlebotomus, Euphlebotomus, Anaphlebotomus,
M a d a p h l e b o t o m u s , L e g e ro m y i a , a n d A b o n n e n c i u s ) ,
Australophlebotomus, Brumptomyia, Oligodontomyia, Sergentomyia
(Sergentomyia, Parrotomyia, Sintonius, Rondanomyia, Vattieromyia,
Trouilletomyia and some ungrouped species), Grassomyia,
Spelaeomyia, Parvidens, Daenemyia, Edentomyia,
Micropygomyia (Silvamyia, Sauromyia, Micropygomyia),
Sciopemyia, Lutzomyia (Helcocyrtomyia, Castromyia,
Tricholateralis, Lutzomyia), Migonemyia (Migonemyia,
Blancasmyia), Pintomyia (Pintomyia, Pifanomyia), Dampfomyia
(Coromyia), Expapillata, Pressatia, Trichopygomyia, Evandromyia
(Aldamyia, Evandromyia, Barrettomyia), Psathyromyia
(Foratiniella, Xiphomyia, Psathyromyia), Viannamyia, Martinsmyia,
Bichromomyia, Psychodopygus, Nyssomyia and Trichophoromyia
regarding the Phlebotominii. We have chosen the following
outgroups: Psychoda sp. Sycorax sp., Nemapalpus spp and
Bruchomyia sp.We sequenced the complete small subunit of the
ribosomal DNA and also several markers of the long subunit (D1, D2
and D8 domains).We compared our results with those previously
published based on morphological characters (Galati 1995, Rispail
and Léger 1998, Rispail and Léger 1998, Galati 2010). Our data will
N° 26 (1) 2015
32
be of help to discuss the systematics of the Phlebotomine sandflies.
DISTRIBUTION, PHYLOGENY AND TAXONOMY OF
SERGENTOMYIA IN ETHIOPIA
RAPID IDENTIFICATION OF PHLEBOTOMINE SAND
FLIES BY MALDI-TOF MASS SPECTROMETRY
Aysheshm Kassahun1, Kristýna Hlavačková1, Vít Dvořák1, Asrat Hailu2,
Alon Warburg3, Jan Votýpka1 and Petr Volf1
Vit Dvořák1, Jérôme Depaquit2, Valentin Pflüger3, Petr Halada4,
Kristýna Hlavačková1, Veronique Lehrter2, Jorian Prudhomme5, Sonja
Zapata6, Petr Volf1, Alexander Mathis7
1
Charles University in Prague, Czech Republic; 2Addis Ababa University, Ethiopia;
Hebrew University of Jerusalem, Israel.
3
ayshek2000@yahoo.com
Keywords: Sergentomyia, leishmaniasis, S. schwetzi
Members of the genus Sergentomyia are dominant sand flies in SubSaharan Africa. Several studies have demonstrated their importance
as vectors of reptilian leishmaniasis caused by Sauroleishmania but
their role, if any, in the epidemiology and transmission of human
leishmaniasis is not well understood. Multiple cases of Sergentomyia
feeding on mammals together with their abundance in the foci of
human leishmaniasis and occasional records of Leishmania parasites
within their bodies detected by microscopy or PCR have led to
speculation on Sergentomyia involvement in the epidemiology of
leishmaniasis. Recently, however, we experimentally demonstrated
that S. schwetzi is refractory to Leishmania parasites infecting
humans. To study the biology, species diversity, phylogeny,
taxonomic status and distribution of Sergentomyia species in Ethiopia,
sand flies have been collected since 2010 using CDC light traps and
sticky traps in endemic areas, mainly in domestic and peri-domestic
habitats (animal shelters, termite mounds, rodent burrows and caves
and acacia and balanite trees). The specimens collected were
identified morphologically and by molecular methods, particularly by
the sequencing of cytochrome C oxidase I and cytochrome B. Seven
Sergentomyia species were found. Males of two morphotypes of S.
schwetzi differed in the orientation of the spines on the style, shape of
the penis and size of the 6th abdominal tergite. Using laboratoryestablished colonies we investigated the effect of ambient temperature
during larval development on the morphological traits of adults. The
genetic variability and proteomic profile were assessed by sequencing
target genes and analysing of the spectrum produced by MALDI-TOF
(Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass
Spectrometry), respectively. Moreover, we investigated the
possibility of parthenogenesis and autogeny in S. schwetzi . This study
was supported by the Bill and Melinda Gates foundation [grant
number OPPGH5336] and Grant Agency of Charles University
[9108/2014].
1)
Department of Parasitology, Faculty of Science, Charles University in Prague,
Prague, Czech Republic. 2) Université de Reims Champagne-Ardenne, ANSES, EA4688
- USC "transmission vectorielle et épidémiosurveillance de maladies parasitaires
(VECPAR)", Reims, France. 3) Mabritec SA, Riehen, Switzerland. 4) Institute of
Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic. 5)
MIVEGEC unit, Centre IRD, Montpellier, France. 6) Instituto de Microbiología,
University San Francisco de Quito, Ecuador. 7) Swiss National Centre for Vector
Entomology, Institute of Parasitology, University of Zurich, Zurich, Switzerland.
icejumper@seznam.cz
Keywords: MALDI-TOF, species identification, protein profiling
Phlebotomine sand flies are vectors of several pathogens causing
diseases in both humans and animals. To elucidate their role in
transmission of these pathogens, proper and conclusive species
identification is crucial. Since morphological determination is based
on minute and sometime dubious characters on their head and
genitalia, which may be altered or absent in field-collected material,
there is a demand for rapid, simple and cost-effective complementary
molecular approaches. Protein profiling using matrix-assisted laser
desorption/ionization time of flight mass spectrometry (MALDITOF MS) was established during the last decade as a routine method
in clinical diagnostics of bacteria and fungi. Currently, applications of
this method on higher organisms including arthropods are emerging.
We demonstrated that MALDI-TOF MS is also feasible for species
determination of phlebotomine sand flies. We established a reference
database of protein spectra of 20 species from the Old World genera
Phlebotomus (subgenera Phlebotomus, Paraphlebotomus,
Larroussius, Adlerius and Euphlebotomus) and Sergentomyia as well
as from four New World genera (Nyssomyia, Psychodopygus,
Psathyromyia and Lutzomyia). Several major vectors of both
cutaneous and visceral human leishmaniases are among analyzed
species. Protein spectra originated from both sexes of laboratoryreared as well as field collected specimens. Best quality spectra were
obtained with frozen specimens, but storage in 70% ethanol is
feasible, and specimens stored for up to 25 years could be identified.
In addition, we evaluated whether mass spectra obtained on the
instruments of the two leading vendors (Bruker Daltonics, Germany;
Shimadzu, Japan) can be used for species identification with the
software of the other instrument. Such cross-applicability would
strongly enhance the value of reference databases established at
different institutions. The present study shows that protein profiling
by MALDI-TOF MS is well applicable on phlebotomine sand flies
and represents an analytical tool which offers high-throughput,
sensitive and reliable species identification which is much desired for
many applications in vector biology.
N° 26 (1) 2015
TWO NEW SPECIES OF THE SUBGENUS
TRANSPHLEBOTOMUS REVEALED BY MOLECULAR
TAXONOMY
DNA SEQUENCE ANALYSIS SUGGESTS THAT CYTBND1 PCR-RFLP MAY NOT BE APPLICABLE TO
S A N D F LY S P E C I E S I D E N T I F I C AT I O N
THROUGHOUT THE MEDITERRANEAN REGION
Vit Dvorak1, Ozge Erisoz Kasap2, Jerome Depaquit3, Bulent Alten2, Jan
Votypka1, Petr Volf1
1
Department of Parasitology, Charles University in Prague, Prague, Czech Republic.
Department of Biology, Hacettepe University, Ankara, Turkey. 3Laboratoire de
Parasitologie, Université de Reims, Ardenne, France. icejumper@seznam.cz
2
Keywords: Transphlebotomus, molecular taxonomy, new species
The subgenus Transphlebotomus comprises only three described
species (P. mascittii, P. canaaniticus, P. economidesi) with markedly
different geographical distribution. While P. mascittii is widely
present in the north of the Mediterranean basin and recently recorded
in Algeria, and stretches up to the northern limits of sand fly presence
in Europe in localities in Belgium, Austria and Germany, P.
canaaniticus and P. economidesi have more restricted distribution.
Due to their similar morphology, proper identification remains
difficult and relies mainly on molecular markers. This subgenus was
long time neglected. The biology of its species is poorly understood,
they are little represented in field collections as some of them are
expected to be cavernicolous and are thought not to be incriminated in
Leishmania parasites transmission, although closely related
subgenera Larroussius and Adlerius harbor several proven vectors of
both cutaneous and visceral leishmaniasis. However, presence of P.
mascittii in several foci of autochtonous canine leishmaniasis in
southern Germany suggests that Transphlebotomus species may be a
vector under certain circumstances. We studied Transphlebotomus
species from Crete, the largest island of the Aegean Archipelago and
from a locality on south-western coast of Anatolia, approximately 500
km away from Crete. To clarify the taxonomic position of the
specimens studied, P. mascittii from Belgium, Germany and France, P.
economidesi from Cyprus and P. canaaniticus from Lebanon were
used as reference sequences. Based on the sequencing analysis of
several genes (cytochrome B, NADH dehydrogenase subunit 4,
cytochrome oxidase I) two new putative species of this subgenus were
found, both of them characterised also by following morphological
characters. One species is described from males only and is mainly
individualised by long third antennal segments coupled with a low
number (13>-20) of coxal setae. The second one is described from
both males and females. The latter are remarkable by their pharyngeal
armature which is Adlerius-like more than Transphlebotomus-like.
Moreover, P. economidesi was found in Turkey sympatrically with
these two species, which is the first record of this species outside
Cyprus. Our findings indicate that the taxonomy of the subgenus
Transphlebotomus should be thoroughly studied by molecular and
morphological methods together with investigations concerning
possible vectorial capacity in Leishmania transmission.
33
Ivonne Pamela Llanes-Acevedo, Carolina Arcones, Rosa Gálvez, Oihane
Martín, Rocío Checa, Ana Montoya, Carmen Chicharro, Silvia
Miguelañez, Susana Cruz, Guadalupe Miró, Israel Cruz
WHO Collaborating Center for Leishmaniasis, Servicio de Parasitología, Centro
Nacional de Microbiología, Instituto de Salud Carlos III. Ctra. Majadahonda-Pozuelo
Km2, 28220 Majadahonda-Madrid, SPAIN Departamento de Sanidad Animal,
Facultad de Veterinaria, Universidad Complutense de Madrid. Avda. Puerta de Hierro
s/n, 28040 Madrid, SPAIN.
cruzi@isciii.es
Keywords: Mediterranean, sandflies, species, cytb-nd1
Molecular methods are increasingly used for both species
identification of sandflies and assessment of their population
structure. In general they are based on DNA sequence analysis of
targets previously amplified by polymerase chain reaction (PCR).
However, this approach requires access to DNA sequence facilities
and in some circumstances is time consuming. Though DNA
sequencing provides the highest degree of information, other
downstream applications of PCR are explored to help in species
identification., It has thus been proposed that the amplification of a
DNA region encompassing partially both the cytochrome-B (cytb)
and the NADH dehydrogenase 1 (nd1) genes, and further digestion
with the restriction enzyme AseI allows for the rapid identification of
the most prevalent species of phlebotomine sandflies in the
Mediterranean region based on their restriction fragment length
polymorphism (RFLP). We sought to validate this methodology using
sandflies collected from different regions in Spain, for which we
amplified the cytb-nd1 target and obtained its DNA sequence. We
completed this data set with DNA sequences available in the
GenBank for species prevalent in the Mediterranean region. In all,
155 DNA sequences from 4 different phlebotomine sandfly species
(Phlebotomus ariasi, P.papatasi, P. perniciosus, and Sergentomyia
minuta) from Spain were generated for this work and added to 146
DNA sequences from 9 different phlebotomine species (P. ariasi, P.
chabaudi, P. neglectus, P. papatasi, P. perfiliewi, P. perniciosus, P.
riouxi, P. sergenti and S. minuta), from 18 different countries
(Afghanistan, Algeria, Cyprus, Egypt, France, Iran, Israel, Italy,
Jordan, Kosovo, Malta, Morocco, Palestine, Portugal, Spain, Syria,
Tunisia, Turkey) obtained from the GenBank. In silico analysis
revealed that this PCR-RFLP method does not provide a unique and
specific profile for each species tested, intraspecific variability occurs
in some of them (P. ariasi, P. papatasi, P. perniciosus and S. minuta),
and there are patterns which are quite similar between different
species (eg. P. chabaudi, P. riouxi, and P. sergenti show a RFLP
pattern similar to intraspecific variants of P. ariasi and P.
perniciosus). In our sample the results obtained by in silico analysis
were further bench tested for confirmation. Though this method has
previously been shown to be useful in Italy, it may not be applicable
throughout theMediterranean region.
Acknowledgement: COLCIENCIAS/COLFUTURO 2012.
N° 26 (1) 2015
MULTILOCUS REAL TIME PCR IDENTIFICATION OF
PHLEBOTOMINE SAND FLIES CAPTURED IN
SOUTHERN ITALY
THE GENUS IDIOPHLEBOTOMUS (DIPTERA;
PSYCHODIDAE)
34
Nicole Léger and Jérôme Depaquit
Reale S, Torina S, Zaffora G, Cosenza M, Pitti R, D'Agostino R, Intile S,
Piazza M, and Vitale F.
Istituto zooprofilattico Sperimentale della Sicilia A. Mirri. Palermo Italy.
Faculté de Pharmacie de Reims France.
nicleger@wanadoo.fr
stefano.reale@izssicilia.it
Keywords: Real Time PCR, Phlebotominae, Leishmania, Diptera
Phlebotomine sand flies (Diptera: Psychodidae) living in
Mediterranean area, are small-sized blood-sucking insects feeding on
a wide range of hosts. They can be vectors of pathogens responsible
for human and animal diseases as leishmaniasis. Out of over 800 sand
fly species that have been described to date, approximately 10% are
proven or suspected vectors of bacteria (e.g., Bartonella
bacilliformis), viruses (e.g., Phlebovirus, Vesiculovirus) as well as
protozoans from the genus Leishmania.. The identification of
phlebotomine sand flies at species level relies on morphology of key
structures including pharynx, spermathecae and cibarium for females
and genitalia for males. Nonetheless, the morphological identification
requires taxonomic expertise and it is time-consuming This study
reports a combined analysis of ribosomal DNA target in phlebotomine
sand flies from the Sicilian Mediterranean region. A 100 bp long
fragments of DNA encompassing a part of ITS2 ribosomal region
were amplified in Real Time test for Phlebotomus perniciosus,
Phlebotomus perfiliewi, Phlebotomus neglectus, Phlebotomus
papatasi, and Sergentomyia minuta, captured at various sites of
southern Italy. Based on high interspecific rDNA difference, a
universal couple of primers and 5 different probes were employed to
reveal the percentage of the different sand fly species in the captures.
Two different PCR mixtures were optimized to detect the cluster
constituted by P. neglectus, P. perfiliewi, P. perniciosus, and the
cluster P. papatasi, S. minuta respectively. On the basis of the obtained
results, we conclude that the test can be used as preliminary approach
to identify the species in captured pool of insects, and to individualize
those that are considerably rare. This study demonstrates the utility of
a multilocus approach to provide a tool for the molecular
identification of the most prevalent phlebotomine sand flies in
southern Europe. Obtained data showed the average percentage for
each species in the catch, indicating the power to resolve the species
mixture collected in the traps used in Leishmania endemic areas.
All the species (16 or 15 ?) of Idiophlebotomus are localized inside
the oriento-australasian area. This genera is the sister group of the
African Spelaeophlebotomus , that is to say that they have a
gondwanian origin and attests of an insular evolution of the
phlebotomine sandflies during the long travel of the future India from
Africa to Asia. Their morphological study is a key for the phylogeny
of the Hertiginii tribe.
ID 25-O Ecology & Epidemiology
SAND FLY SPECIES TRANSMITTING VISCERAL
LEISHMANIASIS AT THE SEMIARID REGION OF
VENEZUELA
Añez Néstor, Rojas Agustina, Yépez José Yancarlos
Universidad de Los Andes, Mérida, Venezuela; Universidad Experimental Francisco
de Miranda, Coro, Venezuela.
nanes@ula.ve
Keywords: Sand fly-species, Visceral-leishmaniasis, Semiarid,
Venezuela
A total of 12 sand fly species were recognized from 6000 specimens
collected during a study carried out in 12 different localities at the
semiarid region of western Venezuela where visceral leishmaniasis
(VL) is endemic. From the total identified specimens, 81% (N=4892)
belong to Lutzomyia longipalpis (15%; N=898) and Lu.evansi (66%;
N=3994). The former, a species incriminated as vector of VL in most
Latin-American countries, and Lu.evansi considered a potential
vector of Leishmania infantum in VL endemic areas from Colombia
and Venezuela. The study also revealed that Lu.evansi was present in
all the 12 sampled localities (100%) while Lu.longipalpis was
collected in 5 of them (41%). In addition, a PCR assay randomly
performed to detect L.infantum from sand fly batches, collected in a
village where active VL cases were detected, revealed infection in 3 of
5 (60%) and 4 of 18 (22%) batches of Lu.evansi and Lu.longipalpis,
respectively. Statistical analysis using the Fisher Exact Test revealed
no significant difference (Pvalue = 0.142) when results for
Leishmania-infection in batches of both sand fly species were
compared. These results suggest that despite the observed abundance
and distribution of Lu.evansi over Lu.longipalpis, both sand fly
species are sympatrically transmitting VL in the semiarid region of
N° 26 (1) 2015
35
western Venezuela.
NATURAL BREEDING SITES OF LUTZOMYIA EVANSI
(DIPTERA: PSYCHODIDAE) IN THE URBAN ZONE OF
OVEJAS, SUCRE - COLOMBIA
LUTZOMYIA FAUNAS (DIPTERA: PSYCHODIDAE) IN
THE EAST SIDE OF THE ANDEAN CORDILLERA IN
CUZCO, PERU
Luis G. Estrada2, Horacio Cadena1, Edgar Ortega2, Luz A Acosta1, Andrés
Vélez-Mira1, Rafael J Vivero1, Eduar E Bejarano2, Sandra Uribe3, Iván D
Vélez1
Perez JE1, D Rado2, D Quispe2, A Quispe2, F Toccas2, M Chacon2, B
Valencia1, A Llanos1, M Quispe2, E Aguilar2, R Pacheco2 & E Ogusuku3
1
PECET (Programa de Estudio y Control de Enfermedades Tropicales) - Universidad
de Antioquia, Medellín, Colombia; 2Grupo de Investigaciones Biomédicas,
Universidad de Sucre, Sincelejo, Colombia; 3Grupo de Investigación en Sistemática
Molecular, Universidad Nacional, Medellín, Colombia.
1
Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana
Cayetano Heredia. 2Instituto Universitario de Enfermedades Tropicales y Biomedicina
del Cuzco. Universidad Nacional San Antonio Abad de Cuzco, Peru. 3Direccion
General de Salud Ambiental, Ministerio de Salud. Lima, Peru.
kikelutz@yahoo.com
luis.estrada@unisucre.edu.co
Keywords: Peru, Lutzomyia, faunas, Cuzco
Keywords: Immature, Phlebotomine, Leishmaniasis, Natural
breeding
The sand flies natural breeding places, despite their potential to assist
the biological control, are poorly understood so far, due to the
difficulties of isolation of immatures stages from the soil where they
occur. The aim of this study was to identify natural breeding sites of
Lutzomyia evansi in three zones of peri-urban area of the municipality
of Ovejas, Sucre – Colombia. This region shows a bimodal climate: a
rainy period from May to November and a dry period from December
to April. The annual precipitation and mean temperature is 1380 mm
and 28°C, respectively, and according to Holdridge the ecological lifezone is Tropical Dry Forest. The study was conducted between March
2013 and February 2014. Each zone consisted of 50 houses and search
of breeding was carried out in the peri and intradomicile of these
homes. Soil samples were taken around the bases of trees, leaf litter,
hollow trees and soil under fallen trees. A total of approximately 503
kg of soil and associated organic matter were processed. Two different
techniques: direct-observation and recovering emerging adults
(incubation cages), were used to verify the presence of immature sand
flies. During the rainy season the largest number of immature sand
flies was recovered mainly in the base of trees. 70 natural breeding
sites positive were identified and distributed as follows: zone1 n = 25;
Zone 2 n =23; zone 3 n = 22. A total of 230 immature forms were
found, which 135 (118 larvae and pupae 17) by direct-observation
while 95 adults emerged by soil incubation. These results showed a
richness of 6 species where Lutzomyia evansi was represented by 118
(51%) followed by Lu. rangeliana with 10 (4%), Lu. cayennensis
cayennensis 7 (3%), Lu. micropyiga 6 (3%), Lu. atroclavata 4 (2%),
and Lu. gomezi with 1 (0.4%). The 55.71% positive natural breeding
sites were Lu. evansi, associated to the base of a shrub called locally as
Uvito (Cordia dentata). 128 immature forms (including 44 exuviae
recovered) that did not reach adulthood will be identified based on
mitochondrial COI gene. This is the first record of peri-urban breeding
sites of Lu. evansi in Colombia. The knowledge about natural breeding
places for sand flies and vector density may represent useful
information for directing efforts at biological control and
consequently control the incidence of disease.
Lutzomyia sandflies are wide distributed in the East side slopes of the
Andean Cordillera of Cuzco Department. The highland vegetation is
dominated by grasses and shrubs, the arboreal vegetation is mainly
the introduced Eucalyptus spp. The lower lands are dominated by
natural tropial forest. The whole area is endemic for bartonellosis
(verruga peruana or Carrion disease) and leishmaniosis
(uta).Objective: To determine the anthropophilic Lutzomyia fauna of
the study area. Sandflies were collected with Shannon traps with
protected human as attractant from 18:00 to 23:00 in the localities
mentioned below. The Lutzomyia faunal composition is notoriously
affected by the altitude, there is one species (L. (Helcocyrtomyia)
peruensis) in Calca, Urubamba, Pisac, Ollantaytambo at around 3000
m of altitude; in San Pedro at 1380 m 5 species were reported (L. (H.)
guderiani 74.9 %). These situations contrast with the reports for the
lower lands, 26 species in Pillcopata at 1000 m (L. (Psychodopygus)
c. carrerai 53.6 %); 37 species in Aguas Calientes at 890 m (L.
(Nyssomyia) y. yuilli 57%); 34 species in Alto Ivochote at 880 m ((L.
(Nyssomyia) y. yuilli 53%); and 29 species in Paratori at 450 m (L.(P.)
llanosmartinsi 72.5 %). Lutzomyia (Helcocyrtomyia) species
predominate in the highest altitudes (>3000m) along the Andean
Cordillera in Peru, this is the case of L. peruensis in Cuzco. The west
side of the Cordillera is dry and arboreal vegetation is restricted to
protected areas in the bottom of some valleys. Lutzomyia fauna in this
area is 3 to 6 species, Lutzomyia peruensis and L verrucarum are the
most common species, both are vectors of bartonellosis and Andean
leishmaniosis (uta).
N° 26 (1) 2015
DIFFERENTIAL EXPOSURE TO SAND FLY BITES IN
SAUDI ARABIA
PREDICTING THE GEOGRAPHICAL DISTRIBUTION
OF LEISHMANIASIS VECTOR SPECIES IN CENTRAL
WEST BRAZIL
Karina Mondragon-Shem1; Waleed Alsalem1,2; Louise Kelly-Hope1,3;
Maha Abdeladhim4; Mohamed Alzahrani2; Ziad Memish2; Jesus
Valenzuela4; Alvaro Acosta-Serrano1, 5
1
Department of Parasitology, Liverpool School of Tropical Medicine, England, UK;
Saudi Ministry of Health, Riyadh, Kingdom of Saudi Arabia; 3Centre for Neglected
Tropical Diseases, Liverpool School of Tropical Medicine, England, UK; 4Vector
Molecular Biology Section, National Institute of Allergy and Infectious Diseases,
National Institutes of Health, Rockville, Maryland, United States of America;
5
Department of Vector Biology, Liverpool School of Tropical Medicine, England, UK.
2
kmshem@liverpool.ac.uk
36
Paulo Silva de Almeida, Alan Sciamarelli, Josué Raizer, Jaqueline
Aparecida Menegatti, Sandra Cristina Negreli Moreira Hermes, Maria
do Socorro Laurentino de Carvalho, Andrey José de Andrade, Rodrigo
Gurgel-Gonçalves
Universidade de Brasília.
rgurgel@unb.br
Keywords: Phlebotominae, ecologic niche modeling, Central West
Brazil, surveillance.
Keywords: Phlebotomus papatasi, biting exposure.
Human antibody response to sandfly saliva can serve as a marker of
exposure to the bite of vector species, and is valuable to assess risk of
disease in different endemic settings. In Saudi Arabia, zoonotic
cutaneous leishmaniasis (ZCL) is caused by Leishmania major and
transmitted by Phlebotomus papatasi, while anthroponotic CL (ACL)
is caused by L. tropica and vectored by Ph. sergenti. Salivary protein
SP32 of Ph. papatasi, the main target of the human immune response
against the saliva of this species, is an ideal biomarker to measure
exposure. The recombinant form of this protein, rPpSP32, allows for
mass screening of sera in endemic areas. In this work, we determined
the level of exposure to Ph. papatasi bites in individuals from CL
endemic areas in Saudi Arabia. We obtained sera from healthy
residents CL as well as patients with active and cured infection, from
CL foci in Al-Ahsa, Al-Madinah and Asir. We measured the levels of
anti-SP32 antibodies by ELISA using rPpSP32. Additionally, we
collected sandflies from the same regions to identify the vector
species present the foci. rPpSP32 was successfully recognized by
human sera from the ZCL endemic areas in Saudi Arabia. We found
higher titres of anti-SP32 antibodies in CL patients than in healthy and
CL-cured individuals in Al-Hasa, while in Al-Madinah there was no
difference between the groups. In both of these ZCL foci Ph. papatasi
was the predominant sand fly species. Interestingly, we also detected a
significant difference between the levels of anti-SP32 antibodies in
patients with papular and nodular lesion types in Al-Madinah. In Asir,
where Ph. sergenti was the most common species, we found low
levels of anti-SP32 antibodies in ACL patients. This revealed a
potential cross-reaction with the Ph. sergenti SP44 PpSP32-like
salivary protein, which was supported by the high similarity with the
SP32 protein sequence. Additionally, we discovered the non-local
workforce in Al Ahsa exhibited higher antibody levels and increased
lesion numbers compared to long-term residents, which will have
important implications for leishmaniasis control programmes in
Saudi Arabia. This is the first study using rPpSP32 protein to assess
biting exposure in CL patients. Cross-reaction with Ph. sergenti
salivary proteins must be kept in mind when evaluating exposure in
areas where both species co-exist. Our results suggest a correlation
between biting exposure, disease outcome and clinical presentation.
Central West Region of Brazil (CWB) comprises the states of Mato
Grosso (MT), Mato Grosso do Sul (MS), Goiás (GO) and the Federal
District (FD). The present study analyzed the geographical
distribution of the main sand fly species associated with leishmaniasis
in CWB and climatic factors associated with its occurrence.
Distributional data of seven sand fly species were obtained by the
entomology laboratories of “Secretarias Estaduais de Saúde”,
museums and literature review between 1978 and 2014. Data were
separated into two sets: one for model calibration (75% of points) and
one for model evaluation (25% of points). Climatic data covered areas
of CWB, resample to a spatial resolution of 5 km. Models were
produced based on eight climatic variables using Maxent. Median
output grids were used as a hypothesis of suitability, and they were
imported into ArcView and DIVA for analysis. Model accuracy was
assessed by examining omission rate associated with test points.
Maxen's jackknife test was used to identify variables that most
influenced models. In total 566 records were analyzed.
Bichromomyia flaviscutellata (n=61), Lutzomyia longipalpis (n=128)
and Nyssomyia whitmani (n=200) had wide distribution in CWB. Lu.
cruzi occurred in 52 municipalities in southwestern and eastern
regions of MT, northern MS and southern GO. Ny. antunesi showed
more restricted distribution in the state of MT and north of MS (n=71).
Ny. intermedia (n=33) and Ny. neivai (n=21) were distributed in the
states of GO, MT and FD. Predictive models indicated areas with high
climatic suitability for Ny. whitmani (southern and northern regions of
MT and MS, central GO), Lu. longipalpis (southwestern MS and
northern MT), Lu. cruzi (southeastern MT, southwestern GO and
northern MS), Bi. flaviscutellata (northern MT), Ny. intermedia
(central GO and southwestern MT), Ny. neivai (southern,
southwestern and eastern MS), and Ny. antunesi (central MT and
eastern GO). The models showed < 1% omission error. The variable
that most influenced models was the temperature in the coldest
months. Moreover, the occurrence of Lu. longipalpis was associated
with dry areas, unlike the models of the other species that were
strongly influenced by the annual precipitation. The results
demonstrated different patterns of geographical distributions for
leishmaniasis vectors in the CWB, which can assist surveillance and
control strategies of this disease.
N° 26 (1) 2015
37
ECOLOGY OF SANDFLIES IN RISK ZONES FOR
TEGUMENTARY LEISHMANIASIS TRANSMISSION
IN SANTARÉM MUNICIPALITY, WESTERN PARÁ
STATE, BRAZIL
PHLEBOTOMUS ORIENTALIS AND THE FALL AND
RISE IN VISCERAL LEISHMANIASIS IN EAST
AFRICA
Dia-Eldin A Elnaiem
Raquel Gonçalves, Daniela Cristina Soares, Ricardo Guimarães, Walter
Souza Santos, Gilberto Cesar de Sousa, Anadeiva Portela Chagas,
Lourdes Maria Garcez
University of Maryland Eastern Shore.
daelnaiem@umes.edu
Instituto Evandro Chagas/SVS/MS and Universidade do Estado do Pará.
luma.garcez@gmail.com
Keywords: Phlebotmus orientalis , East Africa
Keywords: Ecology, sand flies, leishmaniasis, risk zone
Phlebotomus orientalis sand fly is the main vector of Leishmania
donovani in most foci of visceral leishmaniasis (VL, kala azar) in East
Africa. The two other vectors, P. martini and P. celiae, are
responsible of transmission of L. donovani in limited foci in the southeastern region of Ethiopia and the border areas of the Republic of
South Sudan (RSS) and Kenya. Due to marked fluctuations in the
abundance of P. orientalis, East African visceral leishmaniasis is
characterized by periodic outbreaks that result in high morbidity and
mortality rates. The severity of these outbreaks is exacerbated by lack
of knowledge that allows their prediction both in time and space. Our
observations in several areas in Sudan indicate that this vector may
disappear for several years and then erupt in large numbers. While
we believe the cyclical nature of VL and its vector is most likely
related to climatic conditions, little is known about the specific
environmental factors responsible for the epidemics. This is partly
due to lack of reliable surveillance system that provide accurate
estimates of number of cases and abundance of the vector. We
recently embarked on a multicenter research initiative to elucidate the
environmental determinants affecting the fall and rise of VL and use
them to construct a model that can help predict future outbreaks. We
correlated numbers of VL cases with climatic data in eastern Sudan
and RSS. Our results showed that the incidence of the disease is
negatively correlated with the rainfall at the end of the dry season.
These findings correspond closely with our observations on the
ecology of P. orientalis. Future studies will attempt to build Early
Warning System for Visceral Leishmaniasis, based on climate and
other environmental factors.
Tegumentary leishmaniasis (TL) in Brazil used to be near 10 times
more incident than visceral leishmaniasis (VL). The environmental
and socio-economic context influence both in the different endemic
areas. Pará is among the five Brazilian states with the highest
incidence rates of TL over the last years. The municipality of Santarém
is one of the main centres reporting new cases of the disease. We
studied the ecology of sand flies in two spatial circuits (SCs) of TL in
Santarém using the program PAST v.2.03 (PAlentological STatistic)
for calculating ecological indexes: Shannon (H'), Pielou (J) and
Dominance (D). To define the SCs we used geographical coordinates
from transmission and housing locals of patients and the Kernel
density estimator. Thirty-one records selected of a database with 102
TL patients reported in 2010 and 2011 were included. The patients
lived in plateau areas from Santarém, had one or more primary
cutaneous/mucocutaneous lesion and the TL confirmed by
parasitological and molecular diagnosis. Among the etiologic agents
infecting the patients there were Leishmania (Viannia) braziliensis, L.
(V.) shawi, L. (V.) lainsoni, L. (V.) naiffi and one hybrid form of L. (V.)
braziliensis/guyanensis. We identified two SCs of TL in images
classified according the deforestation (2002-2009) and the land uses
(Basin Restoration Program, PRODES – TerraClass Project; National
Institute of Space Research, INPE, Brazil, 2011): Jatobá and CortaCorda. There were captured 417 sand flies (24 species) throughout
two entomological surveys using Shannon and CDC traps. Most were
females (312; 75%) and the sand flies number did not vary in wet
(226/417; 54%) and dry (191/417; 46%) seasons. Nine genus were
identified: Bichromomyia, Evandromyia, Lutzomyia, Micropygomyia,
Nyssomyia, Psathyromyia, Psychodopygus, Sciopemyia,
Trichopygomyia. The ecological indexes varied (H'=0.5-1.72; J=
0.44-0.78; D=0.46-0.91) and confirmed high diversity and the
importance of the vectors Lutzomyia longipalpis (VL) and
Psychodopygus complexus (TL) in Jatobá (urban-rural and
ecotourism area) and Psychodopygus davisi (TL) in Corta-Corda
(rural farm, around a hydroelectric dam). There were less important
the vectors Nyssomyia antunesi, Bichromomyia flaviscutellata and
Nyssomyia anduzei (TL). The results support the Leishmania species
diversity in the patients living in the SCs of TL, representing the
importance of such analysis in entomological surveys.
N° 26 (1) 2015
DISTRIBUTION OF LEISHMANIASIS VECTORS IN
THE MEDITERRANEAN BASIN IN THE LIGHT OF
CLIMATE CHANGE
CHARACTERIZATION OF BREEDING SITES OF
PHLEBOTOMINE SAND FLIES IN THE JUDEAN
DESERT AND NORTH-WEST ETHIOPIA
Bilel Chalghaf, Amine Toumi, Sadok Chlif, Jihene Bettaieb, Mariem
Harrabi, GozeBertin Bénié, Afif Ben Salah
Aviad Moncaz, Oscar Kirstein, Araya Gebresellassie, Wossenseged
Lemma, Solomon Yared, Teshome Gebre-Michael, Asrat Hailu, Moshe
Shenker and Alon Warburg
Institut Pasteur Tunis, Universite de Sherbrooke.
bilel.chalghaf@usherbrooke.ca
Keywords: climate change, ecological niche, modeling, sand flies
Due to climate change, the geographical distribution of sandflies
during the last decades has shifted northward from latitudes below
45○N in south Europe (Martens and McMichael 2002) to latitudes just
above 50○N (Naucke et al., 2008). Recent studies show that some
phlebotomine sandflies were recorded in several parts of Germany
and Belgium (Naucke et al., 2008, Mencke; 2011). In Central Europe,
some autochthone leishmaniasis cases are being recorded in regions
traditionally regarded as leishmaniasis-free countries (Gogoaşe et al.,
2013, Tánczos et al., 2012).In this study we attempted to predict
current distribution of six leishmaniasis vectors in the Mediterranean
basin and forecast species geographical shift under future climate
scenarios using ensemble ecological niche modeling approach.
Species records were obtained from scientific surveys published in
the research literature between 2006 and 2012. A series of climate
metrics, describing temperature and precipitation in the study area
under two climatic scenarios, were obtained from WorldClim
database. We used six individual models contributing to the consensus
model including two regression techniques (Generalized Linear
Models, GLM; Generalized Additive Models, GAM), two machine
learning techniques (Artificial Neural Networks, ANNs; Maximum
Entropy, MaxEnt) and two classification techniques (Surface Range
Envelop, SRE; Random Forest, RF) (why two models from each
category of models?). Alternative formulation: A consensus model
was derived from six varieties of modeling approaches (regression,
machine learning and classification techniques) in order to ensure
valid prediction of distribution of vectors under different climate
scenarios. Model performance was generally high for the included
species with a specificity (True Negative Rate) ranging from 81.03%
to 96.52% (mean=86.94%) and a sensitivity (True Positive Rate)
ranging from 87.93% to 100% (mean=96.98%). Our work evidenced
the hypothesis of the wide spread of Leishamania vectors under
climate change scenarios. All the studied species are prospected to
gain new areas that are actually not suitable for vectors' survival.
Phlebotomine sandflies are prospected to invade extra-Mediterranean
regions especially West.
38
The Kuvin Center for the Study of Infectious and Tropical Diseases, Department of
Microbiology and Molecular Genetics, The Institute of Medical Research IsraelCanada, Faculty of Medicine, The Hebrew University of Jerusalem, Hadassah Medical
School, Jerusalem, Israel.
adi.moncaz@mail.huji.ac.il
Keywords: Phlebotomus vertisols caves trees
Identification and characterization of breeding sites is essential for the
implementation of source-reduction for the control of sand flies and
leishmaniasis. Sand fly breeding sites were identified and
characterized in two foci of leishmaniasis: In the Judean Desert where
the dominant sand fly species that transmits Leishmania tropica is
Phlebotomus sergenti, and in NW Ethiopia where L. donovani is
transmitted by Ph. orientalis. Sand flies emerging from their breeding
sites were trapped by emergence traps (ETs) for 2–8 consecutive
nights. In the Judean desert, Ph. sergenti were trapped emerging from
caves and artificial support walls made of large rocks. . Young
(recently eclosed) male sand flies were found in both habitats. The
temperatures and relative humidity inside caves were stable and soil
samples collected in the cave opening were rich in organic matter. In
Ethiopia Ph. orientalis breeding sites were identified in cracks in
vertisols including fallow fields, dry riverbed/bank and near trees. No
Phlebotomus spp. were trapped in sandy clay loam soil. More Ph.
orientalis were trapped near tree trunks than in open fields but the
difference was insignificant. Large numbers of Sergentomyia spp.
were trapped along with Ph. orientalis mostly outnumbering
Phlebotomus spp. by an order of magnitude. Initial microclimatic
monitoring in the two habitats was conducted at the end of rainy
season, and showed stable subterranean temperatures and RH.
Conditions remained stable through early dry season close to trees but
in open fields temperature fluctuated by 5-6ºC and RH decreased
significantly. Hypoxic conditions evolved during in tree root systems
during the rainy season but oxygen levels rebounded to atmospheric
values once cracks reformed during the dry season. Organic matter
content was high (>5%) in both habitats. Soil pH was moderately
alkaline and electrical conductivity was low in both habitats and
depths. Low oxygen concentrations in soil close to tree root systems
during the rainy season may be caused by microbial metabolism.
Decomposing organic matter, stable temperatures and elevated soil
moisture maintained throughout dry season near the trees at shallow
and deep horizons, and in the open field at deep horizon, probably
N° 26 (1) 2015
39
maintain suitable conditions for sand fly larvae.
F I N E S C A L E A N A LY S E S O F FA C T O R S
CONTRIBUTING TO THE TRANSMISSION OF
VISCERAL LEISHMANIASIS IN NORTH ETHIOPIAN
VILLAGES: AN ECO-EPIDEMIOLOGICAL
APPROACH
SERO-PREVALENCE L. DONOVANI IN LABOUR
MIGRANTS POPULATION AND ENTOMOLOGICAL
RISK FACTORS IN EXTRA-DOMESTIC HABITATS OF
METEMA-HUMERA LOWLANDS - KALA-AZAR
ENDEMIC AREAS IN THE NORTHWEST ETHIOPIA
Oscar David Kirstein, Araya Gebreselassie, Aviad Moncaz, Asrat Hailu,
Teshome Gebre-Michael and Alon Warburg
Wossenseged Lemma1,2, Habte Tekie2, Meshesha Balkew3, Teshome
Gebre-Michael3, Alon Warburg4, Asrat Hailu5
Department of Microbiology & Molecular Genetics, The Institute for Medical Research
Israel-Canada, The Kuvin Centre for the Study of Infectious &Tropical Diseases, The
Hebrew University – Hadassah Medical School, The Hebrew University of Jerusalem.
od.kirstein@mail.huji.ac.i
Keywords: Sand fly, Kala Azar, Epidemiology, Visceral
leishmaniasis
Landscape ecology and landscape epidemiology comprise a set of
tools used for spatial and temporal analyses of vector-borne and other
diseases. In Sudan and Northern Ethiopia, visceral leishmaniasis (VL)
caused by Leishmania donovani, is transmitted by Phlebotomus
orientalis. The epidemiology of VL is the result of a complex matrix of
interactions between the vectors, the pathogens and the hosts with the
environment. This study aims to characterize the risk factors for
contracting VL by gaining a profound understanding of the factors
contributing to its transmission to humans. During 2010 - 2013 a
transverse cohort study was conducted, based on a census comprising
over 11,000 individuals in 18 villages in the Tahtay Adiabo district of
northern Ethiopia. Around 4,900 individuals were screened and tested
for infection with L. donovani. Based on the incidence of VL reported
during the census, the Leishmania infection rates determined by PCR
during the cohort samplings and the environmental and geographical
variables, three villages were selected for an in-depth ecoepidemiological study. In 2014 a nested case-cohort study of ~400
individuals was conducted in the three selected villages, confounder
information available in the previous cohort data-base was used to
select controls that closely match cases in term of sex, age and
geographical location. By analyzing satellite images taken during the
dry season, it was possible to identify that the majority of villages
affected by VL during recent years are inside an area characterized by
vertisols (radius 28 Km). Remote sensing analysis showed high
spectral variability produced by a mix of vertisols, sandstone
outcrops, other sedimentary rocks and varied vegetation land cover.
Entomological studies identified that P. orientalis is closely associated
with vertisol ecosystems rather than specific micro-environments,
being most abundant in cracked black cotton soils. Spatial analysis of
the incidence of VL, show a clustered distribution within populated
areas and micro-clusters of more than one case per house or nearby
house. Houses of patients were most frequently within a few meters of
vertisol fields. These observations may support the hypothesis that the
infection with L. donovani occurs in domestic environments rather
than in the field. The current study is ongoing and more data analysis
will validate or otherwise affect the results presented here.
1
Department of Parasitology, School of Biomedical and Laboratory Sciences, College
of Medicine and Health Sciences, University of Gondar, Gondar, Ethiopia.
2
Department of zoology, Faculty of Life Science, Addis Ababa University, Addis Ababa,
Ethiopia. 3Aklilu Lemma Institute of Pathobiology, Addis Ababa University, Addis
Ababa, Ethiopia. 4Department of Microbiology & Molecular Genetics, The Institute
for Medical Research Israel-Canada, The Kuvin Centre for the Study of Infectious &
Tropical Diseases, The Hebrew University – Hadassah Medical School, The Hebrew
University of Jerusalem, 91120, Israel. 5Department of Microbiology, Immunology &
Parasitology, Faculty of Medicine, Addis Ababa University, Addis Ababa, Ethiopia.
wossensegedlemma@yahoo.com
Keywords: kala-azar infection, labour migrants, risk factors;
Ethiopia
Visceral leishmaniasis (VL) or kala-azar is a systemic disease
transmitted by phlebotomine sand flies and almost always fatal if left
untreated. The aim this study was to evaluate sero prevalence of kalaazar infection and associated risk factors which affect the incidence of
kala-azar. Study on seasonal population dynamics of P.orientalis
were conducted in different villages of Humera before blood
samples was randomly obtained from labour migrants involved in
harvest of sesame for Direct Agglutination Test (DAT) after oral
consents were obtained during October 27 to November 7, 2013. A
structured questionnaire was used to obtain demographic information
and factors that affect the incidence of kala-azar infection in labour
migrants. About 80% of the labour migrants were aged from 15 to 34
years and came from Amhara (75.2%), Tigray (21.2%) and other parts
of Ethiopia (3.6%). Stayed both harvest and weed season in Humera,
Involved in night harvest, Sleep in the farm under Balanites aegiptica
and Knowledge about, at least, a sign or a symptom of kala-azar
were the major factors (P< 0.05) influencing kala-azar infection
(DAT positivity or sero reaction). The total P. orientalis of 7022 (3277
female and 3745 male) were collected from agricultural fields and
thickets in A. seyal in Adebay, Rawyal, Gelanzeraf and Mysegen –
Mehari using 932 CDC traps. The month with the highest mean P.
orientalis/CDC (20.47) abundance was in April. The mean P.
orientalis/CDC during dry season (Nov - April) was highest (Mean ±
Std D =7.33 ± 18.11) compared to May – August weeding (2.05 ±
4.47) and September – October sesame harvest (0.41± 0.77) seasons.
Kala-azar infection in labour migrants seemed to relate with P.
orientalis bite during May – August weeding season compared to
September – October least P. orientalis abundance harvest season.
Peak Kala-azar incidence in permanently settled farmers in villages of
Shiraro also seemed related to P. orientalis bite during May – Augest
weeding season which took 2- 6 months before the
peak
manifestations of the disease in January. Labour migrants are exposed
to kala-azar infection during May - August clearing farm and weeding
season. But, stay during September – October harvest season or
involve in night harvest were not increase the risk of kala-azar
infection as P. orientalis population drops to almost to zero.
N° 26 (1) 2015
40
HOST PREFERENCE PATTERNS OF PHLEBOTOMUS
(LARROUSSIUS) ORIENTALIS PARROT, 1936
(DIPTERA: PSYCHODIDAE) IN THE VISCERAL
LEISHMANIASIS ENDEMIC AREA OF TAHTAY
ADIYABO DISTRICT, NORTHERN ETHIOPIA
BIONOMICS OF PHLEBOTOMINE SANDFLIES IN A
HIGHLAND KALA-AZAR FOCUS IN LIBO-KEMKEM
DISTRICT, NORTHWESTERN ETHIOPIA
Esayas Aklilu, Habte Tekie, Meshesha Balkew, Alon Warburg, Asrat
Hailu, Teshome Gebre-Michael
Araya Gebresilassie1, Ibrahim Abbasi2, Oscar David Kirstein2, Aviad
Moncaz2, Habte Tekie1, Meshesha Balkew3, Alon Warburg2, Asrat Hailu4,
and Teshome Gebre-Michael3
Addis Ababa University.
ak_esas@yahoo.com
1
Department of Zoological Sciences, Addis Ababa University, Addis Ababa, Ethiopia,
Ethiopia; 2Department of Microbiology and Molecular Genetics, The Institute of
Medical Research Israel-Canada The Kuvin Center for the Study of Infectious and
Tropical Diseases, Faculty of Medicine, The Hebrew University, Hadassah Medical
School, Jerusalem, Israel; 3Aklilu Lemma Institute of Pathobiology, Addis Ababa
University, Addis Ababa, Ethiopia; 4Department of Microbiology, Immunology and
Parasitology, Faculty of Medicine, Addis Ababa University, Addis Ababa, Ethiopia.
shambe2006@yahoo.com
Keywords: Bloodmeals, P. orientalis, host preference, Kala-azar
Host choice and feeding success of sand flies are important factors for
understanding the epidemiology and for developing efficient control
strategies for visceral leishmaniasis. The aim of the study was to
determine the host preference patterns of Phlebotomus orientalis from
Tahtay Adiyabo district, Northern Ethiopia. Host attractiveness
experiments were conducted by presenting wild sand flies with a
choice of seven baits (human, cattle, sheep, goat, donkey, dog and
chicken) and control (without bait) in locally designed tent traps. For
blood meal analysis, sand flies were collected from three villages
using CDC light and sticky traps. DNA was extracted from the blood
fed sand flies and PCR amplified for vertebrate-specific cytochrome
(cyt) b region, followed by reverse-line blot (RLB) analysis.
Attraction and feeding success of P. orientalis indicated significant
differences (P < 0.005) between baits. Abundance of P. orientalis on
traps baited with cattle was higher than other animal baits and
controls. Female sand flies were significantly more attracted to
donkey and cattle than human, dog, goat, sheep or chicken-baited
traps. P. orientalis females engorged more frequently on donkey and
cows than they did on other hosts (goat, sheep, dog, and chicken). Out
of 180 P. orientalis tested for the source of bloodmeals by cyt b PCR
and RLB, the blood meals of 135 were successfully identified. P.
orientalis mainly fed on bovine followed by human, goat, sheep and
camel. Mixed blood feeding was also identified in ten female P.
orientalis. The epidemiological significance of these domestic
animals as blood sources for P. orientalis and possible reservoir hosts
of L. donovani should be thoroughly investigated for better
understanding the transmission dynamics of visceral leishmaniasis in
northern Ethiopia.
Keywords: Visceral leishmaniasis, Phlebotomus orientalis,
highland, Ethiopia
The bionomics of phlebotomine sandflies (Diptera: Psychodidae)
were studied for one year from May 2011 to April 2012 in three
villages of Libo-Kemkem district, a highland area where visceral
leishmaniasis has become a major public health problem. The aim of
the present study was to elucidate species composition, resting habits,
seasonal fluctuations of phlebotomine sandflies and incrimination of
the vector. CDC light trap, sticky trap and knockdown space spray
were used for collection of sandflies. Dissection and molecular
approaches were used for Leishmania detection from female
Phlebotomus that were collected by using CDC light traps. A total of
10, 776 sandflies comprising of two species of the genus Phlebotomus
and five species of the genus Sergentomyia were collected.
Phlebotomus orientalis was the predominant species accounting
86.6% of the total sandfly captured. The remaining species in
descending order were Sergentomyia bedfordi group, S.
squamipleuris, S. schwetzi, S. africana, S. clydei and P. rodhaini.
Sandflies were present in most part of the year, except during the main
rainy season (July-September). Phlebotomus orientalis exhibited
two peaks of density, a smaller in January 2012 (6.5flies per trap night
for CDC light trap; 0.98flies/m2/night for STs) and the larger one in
March for STs (1.06flies/m2/night) and April for CDC light trap (18.
7flies per trap night). Abundance of the flies were positively
correlated with average temperature (r=0.53 for CDC; r=0.563 for
STs) and negatively correlated with rainfall (r= -0.272 for CDC; r= 0.171 for STs). The male/female ratio of P. orientalis in the total
captures was 1.87:1.
Behaviorally, P. orientalis exhibited both
exophilic and exophagic. A total of 1060 P. orientalis and 1 P.
rodhaini dissected were found to be negative by microscopic
examination. A similar result was obtained when 247 P. orientalis
females were processed by using Polymerase chain reaction for
detection of the parasite. Although natural infection was not detected
in the current study, P. orientalis is seems to be a vector of VL in this
highland area of the country mainly due to its densely populations and
other circumstantial evidences. Therefore, control of the disease in
this particular area should involve designing of tactics that mainly
target the vector of the disease by considering its seasonal abundance
and behaviors.
N° 26 (1) 2015
SPECIES COMPOSITION, ABUNDANCE AND
SEASONAL DYNAMICS OF PHLEBOTOMUS SPECIES
IN A VISCERAL LEISHMANIASIS ENDEMIC AREA OF
NORTHWEST ETHIOPIA
PHLEBOTOMINE SAND FLIES CROSSING
A N AT O L I A : D O B O R D E R S L I M I T T H E I R
DISTRIBUTION?
Solomon Yared, Meshesha Balkew, Alon Warburg, Asrat Hailu and
Teshome Gebre-Michael
Addis Ababa University.
solyar2005@yahoo.com
Keywords: Visceral leishmaniasis, Phlebotomus spp, P.orientalis,
Habitat preference
Visceral leishmaniasis (VL) is significant public health problem in
northwest Ethiopia particularly in Kafta Humera district. This study
was conceived to investigate the species composition and population
dynamics of sandflies (altitude 500-600masl) in five urban and semiurban area of Kafta Humera district namely Setit Humera, Mykadra,
Rawiyan, Bereket and Adebay. Sandflies were collected for three
nights monthly from May 2011 to April 2012 using CDC light-traps
and sticky traps. Traps were placed within villages, at periphery of
villages and farm fields. Sticky traps were also used for sampling
indoor active sandflies. A total of 13097 sand flies comprising of six
Phlebotomus species of four subgenera were indentified:
Phlebotomus (Larroussius) orientalis, P.(Phlebotomus) papatasi,
P.(Ph.) bergeroti, P.(Ph.) duboscqi, P. (Paraphlebotomus)
alexanderi, and P. (Anaphlebotomus) rodhani. In addition, two
Parvidens (P. lesleyae and P.heischi) species were recorded in the
study area. Among these, P. orientalis was the most predominant
species, accounting for (58.12%) followed by P. papatasi (29.62%), P.
lesleyae (5.61%), P. bergeroti (3.80%), and P. duboscqi (2.07 %), P.
alexandri (0.37%), P. heischi (0.24%) and P. rodhaini (0.18 %). On
average, significantly higher densities of P. orientalis were caught in
the periurban area of Adebay with compared to the urban area of Setit
Humera. Overall, 684 Phlebotomus females were dissected for
detection of Leishmania infection, but none was infected. Significant
positive correlation was found between the monthly abundance of P.
orinetalis and P. papatasi and the monthly averages temperature
(P<0.05) whereas negatively correlated with monthly average rainfall
and relative humidity (P<0.05). Abundance of P. orientalis and P.
papatasi significantly increased during the dry season (January- May)
but decreased in the wet season (June - September) (P<0.05) in the
study areas. With regard to habitat preferences, a large number of
P.orientalis was collected from outdoors especially in the farm fields
with cracked vertisol. In contrast, P. papatasi was highly abundant
indoors as well as outdoors within and the periphery of villages. P.
orientalis is a predominant species and highly abundant during the dry
season and possibly able to transmit VL in the study area. Appropriate
control methods should be designed according to the knowledge of P.
orientalis habitat preferences and seasonal dynamics in relation to
climate condition.
41
O Kasap Erisoz1, A Belen2, C Alkan2, F Gunay1, V Dvorak3, K Ergunay4, S
Aydın5, J Votypka3, A-L Banuls6, R Charrel2, A Özkul7, Y Özbel8, P Volf3, B
Alten1
1
Faculty of Science, Hacettepe University, Department of Biology, Ecology Section,
Beytepe-Ankara-Turkey; 2Unite des Virus Emergents, Marseille Universite, UMR190
Emergence des Pathologies Virales, Faculte de Medecine, Marseille, France;
3
Parasitology Department, Charles University, Faculty of Science, Prague, Czech
Republic; 4Department of Medical Microbiology, Hacettepe University, Faculty of
Medicine, Virology Unit, Sihhiye-Ankara-Turkey; 5Department of Communication
Sciences, Hacettepe University, Faculty of Communication, Beytepe-Ankara-Turkey;
6
IRD (IRD 224-CNRS 5290-UM1-UM2), MIVEGEC, Montpellier, France;
7
Department of Virology, Ankara University, Faculty of Veterinary Medicine, DiskapiAnkara-Turkey; 8Parasitology Department, Ege University, Faculty of Medicine,
Bornova, Izmir, Turkey.
yusuf.ozbel@ege.edu.tr
Keywords: Sandflies, ecology, epidemiology, Anatolia
The term of phlebotomine sand fly was used by Philippo Bonanni in
1691 and then first description was made by Scopoli, under the name
of Bibio papatasi. The name of genus, Phlebotomus, was given in
1840 by Rodani and Berte. Since then, more than 800 species
belonging different genera and subgenera were identified in all over
the world. Their vectorial role for transmitting Leishmania parasites
and phleboviruses were defined at the beginning and mid of the 20th
century, respectively. The Western Palearctic (WP) is composed of
Europe, Middle East and North Africa. In this territory, the
Mediterranean Sea, and the land under the influence of the
Mediterranean Sea is the most important geographical character for
both migration and dispersion of organisms; especially for
invertebrates including sand flies together with pathogen organisms
in it. Anatolia (Asia-Minor) takes place on the crossroads of this area
and has a role as a bridge geographically and biologically.
Leishmaniasis and arbovirus infections due to phleboviruses have
wide geographical distributions in the Western Palearctic, and give
rise to occasional epidemic outbreaks. In numerous countries,
increasing risk factors are making sand fly-borne diseases a major
public and veterinary health problem. Many studies on phylogenetic
relationship among sand fly taxa, their distribution, population
structure and diseases of phlebotomine species have been already
published, but there are still many gaps waiting to be filled up in,
especially, Anatolia. In this point, scientists have to discuss some
deficiencies under cover of geography, history and phylogenetic
studies to understand the mechanisms of distribution of both sand fly
species and their pathogens in Anatolia. In this presentation, updates
in the distribution of sand fly species with state of art maps of EUVBORNET project, possible new species, leishmaniasis and
phleboviruses epidemiology will be discussed with an emphasis on
several studies performed by our group between 2000 and present in
Anatolia.
N° 26 (1) 2015
42
ID 41-O
EPIDEMIOLOGICAL STUDY ON SAND FLIES IN AN
ENDEMIC FOCUS OF CUTANEOUS LEISHMANIASIS,
BUSHEHR CITY IN SOUTHWESTERN IRAN
ECO-EPIDEMIOLOGY OF LEISHMANIASES:
VECTORS AND NEW SCENARIOS OF
TRANSMISSION.
M R Yaghoobi-Ershadi1, M Darvishi1, F Shahbazi1, AA Akhavan1, R
Jafari2, M Khajeian3, H Soleimani4, H Darabi3, M H Arandian2
Jeffrey Jon Shaw
Biomedical Sciences Institute - University of São Paulo, São Paulo - SP – Brasil
School of Public Health, Tehran University of Medical Sciences, P.O.Box 644614155Tehran, Iran
yaghoobi.reza@gmail.com
Keywords: Epidemiology, P. sergenti, L. turanica, Iran
Cutaneous Leishmaniasis (CL) is the most important health problem
in the city of Bushehr, southwestern Iran. The objective of the study
was to determine some ecological aspects of sand flies in the city
during 2010-2011. Sand flies were collected monthly from outdoors
and indoors by sticky traps at four selected districts of the city. They
were also dissected and examined by Nested-PCR for identification of
the parasite during August- September of 2011. A total of 1234 adult
sand flies were collected and 6 species including 3 of Genus
Phlebotomus and 3 of Genus Sergentomyia were identified. Four
species including P. papatasi (3.98 %), P. sergenti (1.14 %), S.
tiberiadis (87.18 %) and S. baghdadis (7.7 %) were found indoors.
Five species including P. papatasi (3.47 %), P. sergenti (3.17 %), P.
alexandri (0.1%), S. tiberiadis (77.85 %) and S. baghdadis (15.41 %)
were collected from outdoors. Sand flies started to appear from March
and disappear at the end of January. There was only one peak in the
density curve in July. The study revealed that S. tiberiadis and S.
baghdadis could enter indoors which 89 % and 81.8 % of them were
found blood-fed respectively. Moreover, P. papatasi, S. tiberiadis and
S. baghdadis were active indoors and outdoors in most months of the
year. Nested-PCR of P. papatasi females were positive against
kinetoplast DNA of L. major and L. turanica and also mixed natural
infections were found by L. gerbilli and L. turanica. Moreover mixed
infections by L. major and L. turanica were observed in this species.
Phlebotomus sergenti females were found infected with DNA of L.
turanica and this the first report of natural infection and detection of
the parasite from this sand fly species in worldwide.
We have no idea of the number of vectors involved in the transmission
of the leishmaniases but in 2010 a WHO publication lists 91 as
suspected or proven. A major difficulty is how to distinguish these
two categories. Attempts to rationalize vector status have been made
by using a variety of epidemiological and parasitological criteria. The
problem is that it is difficult to obtain all these criteria. The following
are those that it is quite feasible to attain: confirmation that the
parasite found in the fly is the same as that found man or suspected
reservoirs; the intensity and localization of the infection; concomitant
ecological distribution with the disease; the degree of anthropophily
and zoophily. It is also important to show that fly transmits the
parasite. During recent years the success rate of experimental
transmissions has improved tremendously but some vectors are
refractory to colonization. This can be overcome by using wild caught
flies. It does not matter whether or not the infections are experimental
or natural. It is against this background that we have to assess vectors
and changes in transmission patterns.
Both the cutaneous and visceral leishmaniases are zoonoses with the
exception of Indian kalazar. Man's relationship to the ecological
niches in which the enzootic cycle occurs is a dynamic process that
depends on a multiple of factors linked to sandfly ecology. Studies on
the ecology of the different vectors are fundamental in understanding
new transmission scenarios.
The major changes in transmission are associated with modifications
of ecological niches that are related to either man's activities or
natural phenomena or a combination of both. The challenge is to
know whether new genotypes are selected by these new conditions or
if they favor the expansion of existing genotypes. Also higher
temperatures increase sandfly metabolism and reduce generation
times that can result in larger vector populations besides possibly
effecting positively or negatively parasite development.
The recent expansion of populations of Lutzomyia longipalpis into
urban areas in Brazil and other Latin American countries has created a
new transmission scenario, but there is evidence that more than one
genotype is involved. There are contradictory studies for risk factors
associated with vectors of the same species. The risk factor for kalazar
in South Asia may be either greater or smaller with the presence of
large animals, such as cows or buffalos. It is not clear why this is but it
may be because Phlebotomus agrentipes populations are modulated
in different ways by the presence of large animals. Both these species
are predominantly domestic and population sizes may also be related
to the quantity of organic material that serves as food for their larvae.
Many more parasites and vectors are linked to cutaneous
leishmaniasis and there are a correspondingly greater numbers of
novel transmission scenarios. L.tropica is typically a parasite of cities
transmitted from man to man by P.sergenti. However, L.killicki,
considered to be a L.tropica variant was found in silvatic areas where
N° 26 (1) 2015
43
the vector is P.sergenti but this fly did not occur in the urban areas of
the same vicinity. In the Americas we also see adaptation of silvatic
flies, such as Nyssomyia whitmani and N.intermedia, to peri-domestic
sites favoring peri-domestic transmission but the sizes of silvatic and
peri-domestic populations is not uniform throughout their
geographical distribution. Potentially molecular markers could help
us understand sandfly habitat and host preferences that are keys in
understanding transmission cycles.
STUDIES ON THE SAND FLY FAUNA (DIPTERA:
PSYCHODIDAE) IN HIGH-TRANSMISSION AREAS
OF VISCERAL LEISHMANIASIS IN NORTH INDIA,
SPECIAL EMPHASIS ON UTTARANCHAL REGION
NS Singh1; Doris Phillips-Singh2; Ipe M Ipe3; Shubham Pandey4; Dinesh
Lal5
EMERGING CUTANEOUS LEISHMANIASIS IN
ISRAEL: A REVIEW OF THE LAST TWO DECADES
Alon Warburg
Department of Microbiology and Molecular Genetics, The Institute of Medical
Research Israel-Canada, The Kuvin Center for the study of Infectious and Tropical
Diseases, The Hebrew University of Jerusalem, Jerusalem, Israel
alonw@ekmd.huji.ac.il
Keywords: Cutaneous Leishmaniasis, Leishmania major, Leishmaia
tropica
In Israel cutaneous leishmaniasis (CL) is caused by Leishmania major
and L. tropica. Until the 1980s the vast majority of CL cases were
caused by L. major and were restricted geographically to the Negev
and Arava Deserts in Southern Israel. However, during the last three
decades CL has expanded its range northwards in hitherto non
endemic areas. The emergent foci of L. tropica in the Galilee region of
Northern Israel, the central Palestinian West Bank and the Judean
Desert affect both urban and rural environments, with most CL cases
residing in peripheral houses bordering open areas. This distribution
suggests that the infected sand flies are arriving from outside the
inhabited area, having acquired the parasites by feeding on infected
rock hyraxes (Procavia capensis), large populations of which are
encroaching upon human habitations in many parts of the country. In
an emerging focus of CL caused by L. major in the Beit She'an Valley
of northern Israel, two rodent species, social voles (Microtus
guentheri) and Tristram's jirds (Meriones tristrami), were
incriminated as new reservoir hosts. The adaptation of L. major to
these hosts has facilitated the emergence of new CL foci where the
main reservoir hosts of the parasite (Psammomys obesus and
Meriones crassus) are absent. In this talk I shall review studies
documenting the emergence of these new foci.
1,2,4,
Sandfly Research Laboratory, Lucknow Christian College Lucknow Christian
College,, Lucknow - INDIA ; 3,5.School of Entomology, St. John's College, Agra-282002
– INDIA
naveendoris@rediffmail.com
Keywords: Phlebotominae, Leishmaniasis, Uttrakhand, India
Visceral Leishmaniasis in the Indian sub-continent, also known as
Kala-azar, is caused by Leishmania donovani transmitted by
Phlebotomus argentipes. This study endeavors to delineate the
suitable locations of Phlebotomus argentipes with relation to
environmental characteristics between endemic and non-endemic
districts in India. The aim of this study was to evaluate the
urbanization of VL vectors in North India, special emphasis on
Uttaranchal Region where VL is endemic. Sandflies were captured
using, light traps along an urban-rural gradient. The traps were
installed outside and inside mud houses at a height of 200 cm. A total
of 2709 sandflies were collected and 9 species were identified. The
predominant species were Sergentomyia punjabaensis (rate of
67.12%) and P. argentipes (0.5%). A total of 1357 females were
dissected and no Leishmania infections were detected. Most of the
sand flies were captured at the border of a forest (76.37%) and no flies
were captured in the urban area, which suggests that transmission is
still restricted to rural sites. However, the fact that a specimen was
collected in an intermediate area indicates that urbanization is a real
possibility and that vector monitoring is important. The present study
should be utilized to make critical decisions for vector surveillance
and controlling Kala-azar disease vectors. At present in India, Bihar
(31 out of 38 districts), West Bengal (10 out of 24 districts), Uttar
Pradesh (4 districts), that border Bihar, and a small portion of the
Jharkhand district are endemic at varying levels. This study provides
the report evaluating the prevalence of Leishmania infection in sand
flies in a region endemic for VL Uttaranchal region in India.
Sergentomyia species are the most common species of sand fly.
Knowledge of the infection rate in female P. argentipes may help in
predicting severity of disease and in vector elimination programs.
N° 26 (1) 2015
INFECTION OF WILD SANDFLIES COLLECTED
FROM DIFFERENT BIO-GEOGRAPHICAL AREAS OF
TUNISIA WITH PHLEBOVIRUSES
INVESTIGATIONS ON SAND FLY BIONOMICS AND
LEISHMANIA NATURAL INFECTIONS IN EASTERN
SICILY, ITALY, WITH PARTICULAR REFERENCE TO
PHLEBOTOMUS SERGENTI
W Fares1, RN Charrel2, K Dachraoui1, L Bichaud2, W Barhoumi1, M
Derbali1, S Cherni1, I Chelbi1, X de Lamballerie2, E Zhioua1
1
Institut Pasteur de Tunis, Laboratory of Vector Ecology, 13 Place Pasteur BP 74, 1002
Tunis, Tunisia. 2Aix Marseille University, IRD French Institute of Research for
Development, EHESP French School of Public Health, EPV UMR_D 190
44
Bongiorno G1, Lisi O2, Severini F1, Vaccalluzzo V2, Khoury C1, Di Muccio
T1, Gradoni L1, Maroli M1, D'Urso V2, Gramiccia M1
1
Unit of Vector-borne Diseases and International Health, Istituto Superiore di Sanità,
Rome; 2Department of Biological, Geological and Environmental Sciences, Section of
Animal Biology “M. La Greca”, University of Catania; Italy
marina.gramiccia@iss.it
Keywords: Sandflies, phlebovirus, emerging, Mediterranean basin.
Keywords: Eastern Sicily, Cutaneous Leishmaniasis, Phlebotomus
perniciosus, Phlebotomus sergenti
An entomological investigation performed in 2013 covering different
bio-geographical areas varying from humid in the north to the arid in
the center showed that sandflies of the subgenus Larroussius including
Phlebotomus perniciosus, Phlebotomus perfiliewi, and Phlebotomus
longicuspis are abundant and widely distributed in Tunisia. A total of
3,992 collected and pooled with up to 30 specimens per pool based on
sex, trapping location and collection date were tested for the presence
of phleboviruses by nested reverse transcriptase polymerase chain
reaction and sequencing. Of a total of 135 pools, 23 were positive,
yielding a minimum infection rate of 0.6%. Phylogenetic analysis
performed using partial amino acid sequence in the polymerase gene
showed that all these phleboviruses were grouped in one cluster
clearly distinct but closely related to Massilia virus and Granada virus.
This putative novel virus, tentatively called Saddaguia virus (SADV),
is widely distributed in Tunisia. Together with Toscana, Punique, and
Utique viruses, SADV is the fourth recognized phlebovirus to be
transmitted by sandflies in Tunisia. The medical and public health
interest of SADV remains to be investigated.
Visceral and cutaneous leishmaniasis (CL) by Leishmania infantum
have long been known to be endemic in Sicily. Catania province, sited
on the eastern coast of the island, represents also an old endemic
territory for Phlebotomus sergenti, the sand fly vector of CL by
L.tropica. Because of the increasing human migration to Sicily and
the related risk for the introduction of exotic Leishmania, an
investigation on sand fly fauna composition, seasonal dynamics and
Leishmania infections was addressed in CL endemic territories. Four
urban (Catania city) and 11 periurban/rural sites, located in a
previously recorded P.sergenti area, were investigated. Sand flies
were collected by sticky traps from April through December and by
CDC light traps from June through October 2013. Live females were
dissected and examined for Leishmania promastigotes identified by
ITS1 PCR- RFLP, whereas dead females were analyzed by
Leishmania nested (n)-PCR.Out of 4105 specimens collected, 2545
(62.0%) were Sergentomyia minuta. Among 1560 Phlebotomus
specimens, P.perniciosus (65.4%) and P.sergenti (20.8%) were the
most prevalent followed by P.papatasi, P.neglectus and P.mascittii.
P.sergenti was confirmed to be endemic, being recorded in 3 urban
and 5 periurban/rural sites of which those located at the slopes of
mount Etna were the most productive. The comparative dynamics of
P.sergenti and P.perniciosus showed that they peaked in different
months, July and August respectively. Females captured alive from
P.sergenti-positive sites (143 P.sergenti and 34 P.perniciosus) were
found negative at dissection. In a P.sergenti-negative site dominated
by P.perniciosus 3/118 (2.5%) specimens of this species were
detected with promastigotes identified as L. infantum. Out of 175
dead females from all sites, 17 (9.7%) were found positive by
Leishmania n-PCR, of which 13 were P.perniciosus and 4 P. sergenti.
While the vector role of P.perniciosus in the L.infantum transmission
was confirmed, that of P.sergenti remains unclear. The detection of
Leishmania DNA in this species suggests an implication in parasite
circulation, however P.sergenti does not seem to play a role as an
active vector of CL, which has been recently confirmed to be caused
by L.infantum in the Catania area. This study was carried out in the
frame of EDENext, a collaborative project of the 7th FP (2011-2014)
funded by the European Commission under the DG Health; Contract
N° 26 (1) 2015
45
Number: 261504.
ID 46-O Vector Competence & Infections
ID 47-O Vector Competence & Infections
N AT U R A L I N F E C T I O N O F L U T Z O M Y I A
C AY E N N E N S I S C AY E N N E N S I S ( D I P T E R A :
PSYCHODIDAE) WITH LEISHMANIA (VIANNIA) SPP.,
POTENTIAL VECTOR IN THE ENZOOTIC CYCLE OF
LEISHMANIASIS IN URBAN AND RURAL HABITATS
OF THE COLOMBIAN CARIBBEAN
PRIORITIES FOR PROGRESSING FROM
ASSESSMENTS OF VECTOR COMPETENCE TO THE
TRANSMISSION MODELLING AND CONTROL OF
LEISHMANIASIS
Paul D Ready
Alveiro Pérez-Doria 1, Margaret Paternina 1, Lily Martinez 1, Luis
Paternina1, Daniel Verbel-Vergara1, Suljey Cochero2, Eduar E. Bejarano1
1
Investigador, Grupo de Investigaciones Biomédicas, Universidad de Sucre, Sincelejo,
Colombia. 2Entomóloga, Departamento Administrativo de Seguridad Social en Salud
de Sucre – DASSSALUD, Sincelejo, Sucre, Colombia.
alveiro_perez@yahoo.com
Keywords: Natural infection, Leishmania (Viannia), Lutzomyia
cayennensis cayennensis, cutaneous leishmaniasis, urban foci.
The first recorded observations in the literature on bloodmeal sources
of Lutzomyia spp. indicate that the members of the subgenus
Micropygomyia Barretto, 1962 feed on reptiles. Since then, and
despite the recent finding of natural infections of Lu. cayennensis
cayennensis with trypanosomatid parasites, the vector potential of this
species has been underestimated due to the low number of insects
captured with CDC light traps in entomological surveys. By contrast,
this was the most abundant species in samples from diurnal resting
sites collected in an autochthonous urban and a rural focus of
cutaneous leishmaniasis in the municipalities of Ovejas and Los
Palmitos respectively, placed in the Montes de María region of the
Sucre, Colombian. This facilitated the search for natural infection with
Leishmania. In all 274 females of the genus Lutzomyia were captured,
dissected and identified in search of trypanosomatid parasites. In
Ovejas 160 females were collected, Lu. micropyga was the most
abundant (42.6%) followed by Lu. evansi (22.8%), Lu. c. cayennensis
(22.2%), Lu. trinidadensis (11.7%) and Lu. panamensis (0.6%). In this
locality natural trypanosomatid infection rates of 4/69 (5.8%) and 2/36
(5.6%) were found in Lu. micropyga and Lu. c. cayennensis,
respectively. In Los Palmitos, the 114 females captured included Lu.
evansi (91.0%), Lu. trinidadensis (0.9%) and Lu. c. cayennensis
(8.0%), the last of these species, with a natural trypanosomatid
infection rate of 2/9 (22.2%). In order to identify the parasites a 350 bp
fragment of Internal Transcript Spacer 1 (ITS1) was amplified and
sequenced. The parasites found in two females of Lu. c. cayennensis
were identified as belonging to the subgenus Leishmania (Viannia), as
suggested by their peripyloric distribution in the sand fly gut.
Consequently in Ovejas Lu. c. cayennensis presented a Le. (Viannia)
spp. infection rate of 2.8%, while in Los Palmitos it was 11.1%. In
conclusion, Lu. c. cayennensis can be naturally infected with parasites
of the subgenus Leishmania (Viannia) and can support their
development, satisfying two of the most important criteria in the
incrimination of a vector. Given the finding of natural infections in
females captured on diurnal resting sites, we suggested that Lu. c.
cayennensis probably acts as a vector in the maintenance of the urban
and rural zoonotic cycles of cutaneous leishmaniasis in the study area
and should be taken into account in the control programs for
leishmaniasis.
Disease Control Department, Faculty of Infectious and Tropical Diseases, London
School of Hygiene and Tropical Medicine, London WC1E 7HT, United Kingdom
Paul.Ready@lshtm.ac.uk
Keywords: vector competence, transmission modelling, risk factors,
vector control
There has been much elegant research on the vectorial competence of
sand flies, which has been aimed at elucidating the factors that permit
Leishmania infections firstly to become established in the abdominal
midgut or hindgut of female flies, and secondly to develop infective
parasite forms and promastigote secretory gel in the thoracic midgut
in order to permit infections to establish in mammalian hosts. There
has also been fascinating research on how female sand flies locate
hosts and physically transmit Leishmania, a process that can be
manipulated by the parasite. In contrast, there has been little research
on modelling the transmission of Leishmania, in order to distinguish
between those sand fly populations that are epidemiologically
important and those that only have the potential to be so. Only the
former need be targeted for control measures. This distinction is
illustrated by Lutzomyia longipalpis: Laboratory populations are
competent vectors of various Leishmania species of the subgenera
Leishmania and Viannia, but wild populations are only usually
associated with the transmission of Leishmania infantum chagasi.
There is a need to prioritise basic research in order to identify sand fly
populations to be targeted for control, and this talk offers suggestions.
Much can be learned from malaria control, including the realization
that the aim of sand fly control should not be to reduce the relative
abundance of a vector species, but to reduce the contact of its blood
feeding females with humans and to reduce the lifespans of the female
flies so that fewer survive long enough to transmit infective parasites.
References: Ready PD. Biology of phlebotomine sand flies as vectors
of disease agents. Annu Rev Entomol. 2013;58:227-250.Ready PD.
Epidemiology of visceral leishmaniasis. Clin Epidemiol. 2014;6:147154.Ready PD, Rogers ME. 2012. Behaviour of sandflies infected
with Leishmania. In Ecology and Control of Vector-Borne Diseases.
Vol. 3: Ecology of Vector-Parasite Interactions, ed. CJM Koenraadt,
W Takken. Wageningen, The Neth.: Wageningen Acad. Publ.
N° 26 (1) 2015
46
ID 48-O Leishmania – Sandfly Interactions
ID 49-O Vector competence & Infections
LEISHMANIA CHAGASI IN ITS NATURAL VECTOR
LUTZOMYIA LONGIPALPIS.
XENODIAGNOSIS OF LEISHMANIA DONOVANI
INFECTIONS IN ASYMPTOMATIC MICE USING
PHLEBOTOMUS ORIENTALIS SAND FLIES
Paulo F P Pimenta, Vanessa C Freitas, Nágila F C Secundino
Laboratory of Medical Entomology, Centro de Pesquisas René Rachou, Fundação
Oswaldo Cruz – FIOCRUZ-MG. Av. Augusto de Lima, 1715 CEP 30190-002 Belo
Horizonte, M.G. Brazil
pimenta@cpqrr.fiocruz.br
Keywords: Leishmania chagasi, sandfly, interaction, Lutzomyia
longipalpis
Studies on Leishmania-sandfly vector interactions are important to
understand processes involved in parasite development and
transmission. Considerable information has been reported mainly for
Old World Leishmania species. In the New World, there is little
detailed information about the development of Leishmania chagasi in
its natural vector Lutzomyia longipalpis. The development of this
parasite was investigated in experimental infection of the vector with
cultured promastigotes and axenic amastigotes obtained by in vitro
transformation. Both parasite forms enable high percentage of
infected sand flies. It was observed a decrease in parasite number
within the gut on the third day after bloodmeal. However, the surviving
promatigotes of both groups were able to multiply and develop in the
gut. The highest density of parasite per gut occurred between six and
seven days and all the promastigote forms were observed in the
parasite life cycle. Experimental infections initiated with
promastigotes and axenic amastigotes showed similar trends of
development and produced infective forms. Therefore, promastigote
infection can be preferably used in experimental infection being less
laboring. For many Leishmania species, the lipophosphoglycan (LPG)
has been suggested to promote the attachment between parasite and
sand fly epithelial midgut. Structural analyses of different Leishmania
species LPG have showed that the polymorphism in theses
glycoconjugates reside in the sugar side chain and cap. The analysis of
strain BH46 side chains demonstrated for the first time a poly
glucosylated LPG (1, 2 and 3 glucose side chains) in L. chagasi. The
LPG importance in survive of this specie in L. longipalpis was
investigated using mutant parasite in LPG biosynthesis. It was
observed reductions in mutant survive or growth prior to excretion
and, in 60 hour, the infection was completely lost. In all natural
Leishmania/vector combinations examined to date, the LPG is
required to binding the parasites to the sand fly midgut to avoid their
loss with the blood meal excretion. Unlike of these observations, we
observed that the LPG synthesis was essential to early survive of the L.
chagasi in L. longipalpis midgut, probably protect the parasites from
enzymatic attack. Morphological aspects of Leishmania/vector
interaction were investigated using light microscopy and scanning and
transmission electron microscopies. Differently of highly specialized
attachment proposed previously to L. chagasi in L. longipalpis, few
parasites were observed with body and flagella in contact with the
epithelial midgut and, not often, with the flagella superficially inserted
between microvilli. In front of no effective vaccine against the disease
and a limited range of drugs to treatment, details in all aspects of
parasite biology are important to formulate new strategies of control
against protozoa and vector.
Jovana Sadlova1, Veronika Hrobarikova1, Jan Votypka1, Alon Warburg2
and Petr Volf1
1
Department of Parasitology, Faculty of Science, Charles University in Prague, 128 44
Prague 2, Czech Republic. 2Department of Microbiology & Molecular Genetics, The
Institute for Medical Research Israel-Canada, The Kuvin Centre for the Study of
Infectious & Tropical Diseases, The Hebrew University- Hadassah Medical School,
The Hebrew University of Jerusalem, 91120, Israel.
sadlovaj@natur.cuni.cz
Keywords: Xenodiagnosis, Visceral leishmaniasis, Asymptomatic
hosts, Reservoir hosts
In endemic areas of visceral leishmaniasis most infected hosts remain
asymptomatic but potentially infectious to biting sand flies.
Therefore, their role in the epidemiology of the disease warrants
intensive study. The best way to test infectiousness of asymptomatic
hosts for sand flies is by xenodiagnosis. However, in humans this test
is problematic for ethical and logistic reasons. In the present study we
established a model using BALB/c mice experimentally infected with
Leishmania donovani to study their infectiousness for Phlebotomus
orientalis, the vector of visceral leishmaniasis in north Ethiopia and
Sudan. To mimic the natural conditions of host infection, mice were
inoculated with parasites isolated from the thoracic midguts of
experimentally infected sand flies with predominance of metacyclic
forms. Although mice did not show any signs of disease, around 20%
of the sand fly females feeding at the site of infection acquired L.
donovani infections. The majority of L. donovani-positive P.
orientalis females (76 %) had heavy infections with the stomodeal
valve fully colonized with attached parasites. Inoculated ears
remained infective for sand flies till the end of the experiment at week
15 post infection. By week 12 post infection females feeding on the
contralateral ear of two mice were also infected, but females feeding
on other parts of the body remained negative. In sacrificed mice the
highest parasite numbers were found in inoculated ears and draining
lymph nodes; infections were detected also in the spleen, liver and
blood, but scarcely in the contralateral ear; and, the other organs
tested were negative. The study showed that asymptomatic Balb/c
mice harbour parasite numbers sufficient to infect P. orientalis. This
finding also encourages searching for more data about the role of
rodents in L. donovani circulation.
N° 26 (1) 2015
ID 50-O Vector competence & Infections
ID 51-O Vector competence & Infection
DEVELOPMENT OF LEISHMANIA PARASITES IN
BITING MIDGES, CULICOIDES NUBECULOSUS AND
CULICOIDES SONORENSIS (DIPTERA:
CERATOPOGONIDAE) AND IMPLICATIONS FOR
SCREENING VECTOR COMPETENCE
THE SAND FLY LUTZOMYIA EVANSI (DIPTERA:
PSYCHODIDAE), AS A PERMISSIVE VECTOR OF
LEISHMANIA INFANTUM AND LE. BRAZILIENSIS IN
BOLÍVAR,
V Seblova-Hrobarikova, J Sadlova, S Carpenter, P. Bates and P Volf
Luis Roberto Romero1, Jorge Luis Rodríguez1, Alveiro Pérez-Doria1,
José G. Vergara, Ana Montesino1, Lily Paola Martínez1, Luis Cortés2,
Suljey Cochero 1,3 & Eduar Elías Bejarano1
Department of Parasitology, Charles University in Prague, Faculty of Science, Prague
2, 128 44, Czech Republic
Vera@seznam.cz
47
COLOMBIA
1
Grupo de Investigaciones Biomédicas, Universidad de Sucre, Sincelejo, Colombia;
Secretaría de Salud Departamental de Bolívar, Cartagena, Colombia; 3Secretaría de
Salud Departamental de Sucre, Sincelejo, Colombia.
2
Keywords: biting midges, Leishmania parasites, vector competence
Biting midges of the genus Forcipomyia (Diptera: Ceratopogonidae)
have recently been implicated as vectors of kinetoplastid parasites in
the Leishmania enriettii complex. Here, we assessed susceptibility of
two colonized midges, Culicoides nubeculosus and C. sonorensis, to
Leishmania infection. Midges C. nubeculosus and C. sonorensis were
experimentally infected with two strains from the L. enriettii complex,
L. australiensis originally from red kangaroos (LV756) and L. enriettii
from guinea pigs (LV90), and two Leishmania species infecting
humans, L. major (FVI) and L. infantum (CUK3). A permissive
species of sand fly, Lutzomyia longipalpis, was used as a control.
Engorged females were microscopically examined at different time
points post-bloodmeal to assess the localization of Leishmania
promastigotes in the gut. In C. nubeculosus, all four tested Leishmania
strains produced only light infection in the abdominal midgut with
presence of long nectomonads. In contrast, C. sonorensis was able to
support late-stage development of both strains from the L. enriettii
complex; in 20% of females, parasites migrated anteriorly and
colonized the stomodeal valve, which is one prerequistite for
subsequent transmission. Similarly, L. longipalpis was susceptible to
parasites from the L. enriettii complex but only to strain LV756. In
summary, we showed that the palaearctic species of biting midge C.
nubeculosus is not susceptible to any Leishmania species tested. In
contrast, L. longipalpis and C. sonorensis supported late stage lifecycle development of parasites from the L. enriettii complex, which is
promising for their transmission potential. However, detailed
transmission experiments are needed to avoid mistaken speculations
about vector competence of these species for members of the
Leishmania enriettii complex.
luisro987@gmail.com
Keywords: Lutzomyia evansi, permissive vector, Montes de María.
Los Montes de María on the Caribbean coast of Colombia is
considered to be a mixed focus of leishmaniasis transmission due to
the presence of cases of both the cutaneous and visceral forms of the
disease. In this area of the country Leishmania infantum, the
aetiological agent of visceral leishmaniasis, is known to be
transmitted by Lutzomyia evansi. However little is known about the
vectors of Le. braziliensis and Le. guyanensis, aetiological agents of
cutaneous leishmaniasis, the clinical form with the greatest number of
human cases. This study focused on the detection and identification of
trypanosomatids in phlebotomines from a municipality of the Montes
de María region. Four collections were made with Shannon trap and
by sampling from diurnal resting sites with an electronic aspirator in
the Las Margaritas neighbourhood of El Carmen de Bolívar
(9°43'58,808''N; 75°07'32,962''W), during two rainy and two dry
seasons in 2012-2013. The guts of female phlebotomines were
extracted and examined under the microscope for trypanosomatid
parasites, as well as the spermathecae for taxonomic determination of
the species. The parasites found in the gut were genotyped using PCR
and sequencing in tandem of a region of the cytochrome b gene and
another of subtelomeric repetitions. In all 2509 sand fly specimens
were collected, of which 88.9% were Lu. evansi, 2.0% Lu.
cayennensis cayennensis, 0.7% Lu. gomezi and 0.2% Lu.
panamensis. Two specimens of Lu. evansi were found naturally
infected with trypanosomatids. One presented a suprapyloric
distribution of the parasites, identified by molecular techniques as Le.
infantum, while in the other specimen the trypanosomatids were
distributed peripylorically and found to be Le. braziliensis. These
findings demonstrate the permissive character of Lu. evansi as a
vector of two Leishmania species and its importance in the
epidemiological cycle of cutaneous leishmaniasis in the Colombian
Caribbean.
N° 26 (1) 2015
48
ID 52-O Vector competence & Infection
ID 53-O Vector-Parasite-Host Interactions
EXPERIMENTAL SUSCEPTIBILITY OF LUTZOMYIA
(L.) LONGIPALPIS TO DIFFERENT SPECIES OF
LEISHMANIA
THE PROSPECT FOR A SAND FLY SALIVA-BASED
VACCINE AGAINST
LEISHMANIASIS IN HUMANS
Ana Clara Araújo Machado Pires, Paulo Filemon Paolucci Pimenta and
Nágila Francinete Costa Secundino
Shaden Kamhawi, Fabiano Oliveira and Jesus Valenzuela
Laboratory of Medical Entomology of the Centro de Pesquisas René Rachou,
Fundação Oswaldo Cruz – MG
Vector Molecular Biology Section, Laboratory of Malaria and Vector Research,
National Institute of Allergy and Infectious Diseases, National Institutes of Health.NIAID, NIH-
ana.clara.pires@hotmail.com
Keywords: Susceptibility, Lutzomyia (L.) longipalpis, Leishmania
In order to better understand the epidemiology of leishmaniasis and
the parasite transmission it is important to develop studies on the
Leishmania-vector interaction. The knowledge of the process of the
sand fly-parasite interaction can contribute for new control strategies
of leishmaniasis, a complex disease that until now has no effective
vaccine and limited range of drugs for treatment. In nature, some
sandfly species show remarkable specificity to transmit an exclusive
Leishmania species (non-permissive vector), while others can be
vectors of more than one parasite species (permissive vector).
However, in the laboratory, some sandfly like Lutzomyia (L.)
longipalpis, exclusive vector of L. (L.) chagasi, can be infected
experimentally by different parasite species. One important step for
Leishmania to establish infection in sandfly vector is to be capable to
attach to the midgut epithelium, which is known to be dependent of
LPG molecules in non-permissive vectors. However, the parasite
attachment in the permissive vectors appears to be different and a nonLPG dependent mechanism. Here, we describe details of the
development of four distinct species of Leishmania: Leishmania (L)
major, L. (L.) amazonensis, L. (Viannia) braziliensis and L. (L.)
chagasi infantum in the sandfly Lu. (L.) longipalpis, a lab-permissive
vector. Experimental infections using membrane feeding device were
conducted using three parasite doses (4x107, 2x107 and 1x107) mixed
with mouse blood. On day 6 after the complete digestion of the
infective blood meals, it was observed that L. (L.) chagasi infantum, L.
(L.) major and L. (L.) amazonensis, in all tested parasite doses,
infected the Lu. (L.) longipalpis. However, in L. (V.) braziliensis only
the highest dose (4x107) was able to develop infection in the vector but
with a low average of infection rate (1,000 parasites/sandfly). In
contrast, even with the lowest parasite dose, L. (L.) amazonensis and
L. (L.) major showed an average of 7,000 parasites/sandfly, while L.
(L.) chagasi showed an average of 10,000 parasites/sandfly. Although
it is well known that Lu (L.) longipalpis in the nature can only transmit
L. (L.) chagasi it is still need further studies to prove that this vector
can transmit other Leishmania species.
Financial SUPPORT: FAPEMIG, FIOCRUZ, CNPq and Capes.
skamhawi@niaid.nih.gov
Keywords: Vector saliva, DTH response, Th1 immune response,
Leishmania vaccine
To date, a human vaccine against any form of leishmaniasis is not
available. Immunity to saliva or distinct salivary proteins has a
profound impact on host immunity to the Leishmania parasites. In
most cases, saliva-specific immunity drives a protective immune
response against Leishmania. Protective molecules have mostly
shared a similar property, the induction of a delayed-type
hypersensitivity (DTH) response biased towards a Th1 profile. This
protection has been observed in rodents and non-human primates and
accelerates the development of a robust Leishmania-specific immune
response with little pathology. Importantly, immunity to a TH1-DTHinducing salivary protein has protected against both cutaneous and
visceral leishmaniases suggesting that it is acts on parasites in the skin
soon after an infected bite when they are at their most vulnerable. The
strong protection mediated by immunity to saliva in various models
argues for their inclusion in a leishmaniasis vaccine. The
opportunities and challenges facing a combination salivaLeishmania vaccine will be discussed.
N° 26 (1) 2015
CHARACTERIZATION OF LUTZOMYIA LONGIPALPIS
IMMUNE RESPONSES WITH EMPHASIS IN
LEISHMANIA INFECTION
P R O M A S T I G O T E S E C R E TO RY G E L F R O M
INFECTED SAND FLIES SUBVERTS THE WOUND
HEALING RESPONSE IN AN INSULIN GROWTH
FACTOR1-DEPENDENT MANNER
Pitaluga AN, Tinoco B, Di Blasi T, Silva AM, Tempone A, Traub-Csekö
YM
Instituto Oswaldo Cruz, Fiocruz
ytraub@ioc.fiocruz.br
49
Matthew E Rogers, Emilie Giraud, Rod J Dillon, Paul A Bates, Tamsyn
Derrick, Oihane Martin, Ingrid Műller
London School of Hygiene and Tropical Medicine
matthew.rogers@lshtm.ac.uk
Keywords: Lutzomyia longipalpis, innate immunity
Keywords: Leishmania transmission, PSG, IGF1
Lutzomyia longipalpis is the major vector for visceral leishmaniasis in
Brazil. We are studying immune responses both in the insect vector
and in the L. longipalpis LL5 embryonic cell line. Insect innate
immune response pathways are Toll, IMD and JAK-STAT. We have
identified an IMD and Toll response in LL5 cells through the silencing
of the respective repressor genes caspar and cactus. This silencing
produced the expected effect of increasing the expression of the
antimicrobial peptides (AMPs) defensin and cecropin, thus
establishing LL5 cells as a valid model to study sandfly immunity. We
also investigated de effect of silencing these repressors in female adult
insects. We have previously determined a role for the IMD pathway in
the vector infection by Leishmania through the silencing of the
repressor caspar, which decreased infection. We now silenced cactus
and, surprisingly, as opposed to LL5 cells, this led to the decrease of
AMPs production. As a possible explanation, we verified that WntD,
an inhibitor of the Toll pathway identified in Drosophila, had
increased expression upon cactus silencing. This might be related to
the preservation of the insect microbiota, which was actually
decreased when cactus was silenced. We have also studied the effects
of Leishmania infection on L. longipalpis. Infection caused an early
increase of cactus expression followed by a return to normal levels
that was accompanied by a higher expression of AMPs. Interestingly
the infection by Leishmania also caused an early growth of the
microbiota, probably related to the increase of cactus expression. We
were intrigued by the apparent capacity of Leishmania to increase the
expression of cactus and thus inhibit the Toll pathway. It is known that
Leishmania Gp63 activates a macrophage tyrosine phosphatase
(SHP-1), capable of inhibiting the Toll and Jak/STAT pathways. L.
longipalpis has a homoloque of SHP-1, which has increased
expression during L. longipalpis infection by L. i. chagasi, indicating
the possible conservation of a mechanism of Leishmania infection
control in mammals and insects.
In the sand fly midgut secreted proteophosphoglycans from
Leishmania parasites form a biological plug known as the
promastigote secretory gel (PSG), which facilitates the regurgitative
mode of transmission and infection of the mammalian host. Although
the disease-exacerbating properties of PSG have been related to
macrophage alternative activation the underlying signalling
pathways remain unknown. Affymetrix Mouse GeneChip and
Pathway analysis revealed differential expression of 7,927 transcripts
(fold change >1.5, p<0.05) to Leishmania mexicana PSG in BALB/c
mouse ears (27% of the mouse genome). These transcripts were
transiently expressed during the first 6 hours and enriched for
pathways involved in inflammation, cell proliferation, fibrosis,
epithelial cell differentiation and wound remodelling. We found that
L. mexicana PSG significantly accelerated wound healing in vitro
and in vivo; which was associated with an early up-regulation of
transcripts involved in inflammation (IL-1a, IL-6, TNFa) and
inflammatory cell recruitment (CCL2, CCL3, CCL4, CXCL2),
followed 6 days later by enhanced expression of transcripts
associated with epithelial cell proliferation, fibroplasia and fibrosis
(FGFR2, EGF, EGFR, IGF1). Antibody blockade of IGF1 ablated the
gel's ability to promote wound closure in BALB/c mouse ears and
significantly reduced the virulence of L. mexicana infection delivered
by an individual infected sand fly bite. Dermal macrophages recruited
to air pouches on the backs of mice revealed that IGF1 was pivotal to
the PSG's ability to promote macrophage alterative activation and L.
mexicana infection.
N° 26 (1) 2015
V E C TO R - T R A N S M I S S I O N O F L E I S H M A N I A
DONOVANI TRIGGERS A DISTINCT IMMUNE
RESPONSE THAT FAVORS PARASITE SURVIVAL AND
DISSEMINATION
CYTOKINE LEVELS IN RESPONSE TO SALIVARY
GLAND EXTRACTS AND TO LEISHMANIA MAJOR
PARASITES IN INDIVIDUALS LIVING IN ENDEMIC
A R E A S F O R Z O O N O T I C C U TA N E O U S
LEISHMANIASIS IN TUNISIA
Ranadhir Dey1, Hamide Aslan2, Amritanshu Joshi, Claudio Meneses2,
Pradeep K. Dagur3, John Philip McCoy3, Robert Duncan1, Jesus G.
Valenzuela2, Hira L. Nakhasi1* and Shaden Kamhawi2*
1
National Institute of Allergy and Infectious Diseases, National Institutes of Health.NIAID, NIH. 2Vector Biology, National Institute of Allergy and Infectious Diseases,
Baltimore, USA;
skamhawi@niaid.nih.gov
50
Wafa Rebai-Kammoun, Ikbel Naouar, Darragh Duffy, Valentina Libri,
Milena Hasan, Amine Toumi, Afif B. Salah, Matthew Albert, Hechmi
Louzir, Amel Meddeb-Garnaoui
Laboratory of Medical Parasitology, Biotechnology and Biomolecules, Institut
Pasteur de Tunis; Tunisia;Laboratory of Transmission Control and Immunobiology of
Infection, Institut Pasteur de Tunis; Tunisia; Laboratory of Dendritic Cell Biology,
Department of Immunology, Institut Pasteur, Paris, France; : Center for Human
Immunology, Institut Pasteur, Paris, France
kammoun.wafa@gmail.com
Keywords: Vector-tranmission, Leishmania donovani, immune
response, visceral leishmaniasis
Keywords: Phlebotomus papatasi, Salivary gland extract,
Leishmania, immune response
Visceral leishmaniasis (VL) caused by Leishmania donovani is
prevalent in the Indian subcontinent, South Asia, and East Africa and
is considered mostly an anthroponosis. Most VL models of infection
involve intravenous, intracardiac, or intraperitoneal injection of
106–108 parasites into laboratory animals. These routes bypass the
skin, the natural delivery site of the parasites by sand fly bites. For
cutaneous leishmaniasis, natural transmission by vector bites revealed
important distinctions in disease initiation and progression and
abrogated protection observed in vaccinated animals following needle
challenge. Here, we infected BALB/c mice via bites of L. donovaniinfected Lutzomyia longipalpis sand flies and compared disease
development to intravenous (i.v.) or intradermal (i.d.) needle-initiated
infections. The mice infected via sand fly bites- developed
Leishmania-specific antibodies that correlated with dissemination of
parasites into the spleen and liver; by weeks 20-30 post-infection,
most mice cleared liver parasites but displayed a slight increase in the
splenic parasite burden. Following i.v.-injection, the parasites
persisted in both the spleen and liver at 20-30 weeks post-challenge
while i.d.-injected parasites failed to disseminate to visceral organs.
The immune response to vector-initiated infections was silent with
splenic CD4+ and CD8+ T cells producing less IFN-g and TNF-a at 5
and 30 weeks post-infection but similar levels of IL-10 compared to
the i.v.-initiated infection. In the skin, vector-initiated infection
induced an acute pro-inflammatory response (IFN-g, IL1-b, TNF-a
and NOS2) 3h post-transmission that was not observed following i.d.injection of parasites. This response subsided at 6h to be replaced by a
persistent recruitment of neutrophils and alternately activated
monocytes to the bite site characterized by a sustained induction of IL10, MCP1, Ym1 and ILRN1 up to 18h post-infection. Altogether,
vector-transmission induces an acute pro-inflammatory response
followed by a persistent and immunologically quiet cellular
infiltration at the bite site that is distinct from the immune response to
i.v.- and i.d.-initiated infections. This immunologically silent
environment may facilitate the dissemination and survival of L.
donovani in its mammalian hosts.
We analyzed cellular proliferative and IFN-γ and IL-10 responses
(ELISA) after stimulation by salivary gland extracts (SGE) from
Phlebotomus papatasi, vector of L. major, in 378 individuals living in
endemic areas for cutaneous leishmaniasis (CL) due to L. major.
These were subdivided according to the leishmanin skin test (LST)
response and the presence of scars: healed (HCL: LST+/Scars+),
asymptomatics (As: LST+/Scars-) and naïves (LST-/Scars-). A
significant IFN-γ response to SGE was only observed in naive
individuals. IL10 was not detected in response to SGE in all analyzed
groups. To further analyze cytokine response to SGE, we used
Luminex technology to measure IL-1b, IL-2, IL-5, IL-6, GM-CSF,
IL-13, TNF-a, IL-12, IL-18, IFN-γ and IL-10 levels in supernatants
from SGE-stimulated PBMC derived from HCL (n=17), As (n=23)
and Naives (n=18). HCL and As individuals were further selected on
the basis of a strong positive IFN-γ responses to L. major parasites.
Our results showed that all cytokines analyzed were not detected after
SGE stimulation in these groups. The significant IFN-γ response
detected by ELISA could be explained by the larger number of
individuals tested. Interestingly, a clinical follow up of these
individuals allowing to detect new CL case have shown that no new
CL cases were observed among the naïve individuals, who showed
positive anti-SGE proliferative responses unlike those with negative
proliferative responses (6 new cases). These results suggest that
cellular responses developed against salivary components in the
absence of parasites may induce, under certain conditions, a
protection against subsequent Leishmania infection. We also
evaluated cytokine levels in supernatants from L. major-stimulated
PBMC. A significant response was detected for IFN-γ, IL-1b, IL-2,
IL-5, IL-10, GM-CSF, IL-13, TNF-a, IL-12 and IL-18 in HCL and As
individuals. Interestingly, GM-CSF levels were significantly higher
in HCL compared to As individuals. GM-CSF is an important
hematopoietic growth factor and immune modulator. Some data
support a protective effect of GM-CSF on the course of Leishmania
infection. Analysis of these cytokine profiles may allow to identify
biomarkers associated with disease outcome and to better understand
the mechanisms of immune response against leishmaniasis.
N° 26 (1) 2015
P R O S P E C T I V E S T U D Y O F A C O H O RT O F
INDIVIDUALS AT RISK OF ZOONOTIC CUTANEOUS
LEISHMANIASIS IN TUNISIA TO EVALUATE
CLINICAL AND IMMUNOLOGICAL PARAMETERS
AND IDENTIFY CORRELATES OF PROTECTION.
THE POSSIBLE ROLE OF A FLAGELLAR PROTEIN
FROM LEISHMANIA SP. (FLAG) IN THE PARASITE SAND FLY INTERACTION
Wafa Rebai Kammoun, Ikbel Naouar, Amel Meddeb-Garnaoui, Amine
Toumi, Nabil Belhadj Hamida, Afif B Salah, Hechmi Louzir
Laboratory of Medical Parasitology,Biotechnology and Biomolecules, Institut Pasteur
de Tunis, Tunisia ;Laboratory of Transmission Control and Immunobiology of
Infection, Institut Pasteur de Tunis, Tunisia
51
Tatiana Di-Blasi1; Antonio Jorge Tempone1; Erich Loza Telleria1;
Fabricio K Marchini2; José Marcelo Ramalho-Ortigão3; Yara Maria
Traub-Csekö1
1
Instituto Oswaldo Cruz - Fiocruz, Rio de Janeiro - RJ - Brazil; 2Instituto Carlos
Chagas - Fiocruz, Curitiba - PR - Brazil; 3Kansas State University, Manhattan - KS United States of America.
tdiblasi@ioc.fiocruz.br
kammoun.wafa@gmail.com
Keywords: Sand fly, Leishmania, Interaction, Flagellum
Keywords: Zoonotic cutaneous leishmaniasis, endemic areas,
Leishmanin Skin Test, correlates of protection
The Leishmaniases constitutes a heterogeneous group of diseases that
affect millions of people and are caused by various Leishmania (L.)
species. Leishmania major is the causative agent of zoonotic
cutaneous leishmaniasis (ZCL). The aim of this study was to establish
and follow a prospective cohort of individuals at risk, living in
endemic areas of L. major, to evaluate markers of exposure to
Leishmania infection. In 790 individuals in 5 different endemic areas
for L. major, we observed: positive Leishmanin Skin Test (LST):431
individuals and 190 had leishmaniasis typical scars. We observed an
increase of GrB, IFNγ and IL10 production with the age of the study
subjects (p=0.002, p=0.001 and p=0.019 respectively) and with the
size of LST reactions for GrB and IFNγ (p<0.001). Moreover, a
positive correlation between LST and both IFNγ and GrB production
was observed. When considering the foci type, individuals living in
Mnara, the oldest focus, responded with the highest levels of IFNγ,
GrB but also IL10 which seems to be an indicator of exposure to
Leishmania infection (p<0.001). We also observed a strong
correlation between IFNγ and GrB (rho=0.80). After a transmission
season, 29 ZCL cases were observed. IFNγ/IL10 and GrB/IL10 ratios
were also calculated in order to investigate the potential correlation of
these ratios with other with the resistance against ZCL. Interestingly,
the majority of individuals who developed ZCL lesions had an
IFNγ/IL10 ratios <10 (the lowest ratios), when only 7% of emerging
ZCL cases had very high ratios. A significant decrease of ZCL cases is
observed with the increase of IFNγ/IL10 ratio.
Leishmaniasis diseases are caused by Leishmania protozoans and
transmitted to the vertebrate host mainly through the bite of
Lutzomyia (New World) or Phlebotomus (Old World) sand fly
species. These parasites are ingested by the sand fly during blood
feeding and adhere to the midgut epithelium through the flagellum
which is an important step to avoid parasite excretion after blood
digestion. Previous studies identified a Leishmania flagellar protein
called FLAG potentially involved in the parasite adhesion to the sand
fly gut. To verify the role of FLAG in the midgut adhesion process, we
performed ex vivo and in vivo inhibition assays using an anti-FLAG
monoclonal antibody. The experiments were performed with the pairs
Leishmania infantum chagasi - Lutzomyia longipalpis, and
Leishmania major - Phlebotomus papatasi, in which the parasites
were previously incubated with the anti-FLAG antibody. The results
obtained from both ex vivo and in vivo assays showed that only the
pair L. major and P. papatasi presented a significant reduction in the
number of parasites inside the gut when they were pre-incubated antiFLAG. In parallel, we performed pull down experiments in order to
identify L. longipalpis and P. papatasi molecules that could interact
with FLAG. For this purpose, we used extracts from L. longipalpis
(LL5) and P. papatasi (PP9) cultured embryonic cells. In the
experiments with LL5 extracts we observed only nonspecific
bindings to the fusion protein or to the resin and no specific binding to
FLAG. However, in the assays with PP9 extract we could identify
possible FLAG binding molecules that were characterized through
mass spectrometry. Among the molecules identified through the
sequencing there are: PDGF- and VEGF-receptor related protein
known as having a role in oogenesis and cell migration in Drosophila
melanosgater. Another molecule identified was a fas-associated
protein related with immunological response in Aedes aegypti. The
interaction between parasite and its vector is a key step involved in the
establishment of leishmania in the sand fly. Therefore the study of
molecules involved in this interaction can contribute to develop new
strategies to block parasite transmission. Financial support: CNPq;
FIOCRUZ.
N° 26 (1) 2015
52
sand fly feeding on avian hosts is not harmful to Leishmania
parasites.
ID 61-O Genome & Genetics
THE EFFECT OF TEMPERATURE AND AVIAN BLOOD
ON LEISHMANIA DEVELOPMENT IN SAND FLIES
TWO SAND FLY GENOMES: PHLEBOTOMUS
PAPATASI AND LUTZOMYIA LONGIPALPIS
Votypka J, Pruzinova K, Hlavacova J, Volf P
Department of Parasitology, Faculty of Science, Charles University, Prague, Czech
Republic
vapid@natur.cuni.cz
Mary Ann McDowell and the Sand Fly Genome Consortium
Eck Institute for Global Health, Department of Biological Sciences, University of
Notre Dame, USA
mcdowell.11@nd.edu
Keywords: temperature, blood, leishmania development
Keywords: Lutzomyia longipalpis, Phlebotomus papatasi, genome
Ambient temperature is one of external factors affecting pathogen
transmission by haematophagous arthropods. Recent findings of new
leishmaniasis foci in Latin America and Europe and the spread of the
disease connected with the climate changes require more data on the
effect of temperature on Leishmania development in sand flies.
Therefore, the development of three important Leishmania species (L.
infantum, L. braziliensis and L. peruviana) in two sand fly vectors
(Phlebotomus perniciosus and Lutzomyia longipalpis) was compared
at two different ambient temperatures (20°C and 26°C). At various
intervals post-blood meal, infected females were examined under a
light microscope and qPCR was used for parasite quantification.
Leishmania peruviana, a mountain species living in lower
temperatures, thrived well in Lu. longipalpis females kept at 20°C,
while at 26°C almost all infections were lost due to fast digestion and
early defecation of the blood remnants. On the other hand, L.
braziliensis and L. infantum developed well in sand fly females kept at
both temperatures tested. The ability to develop at low ambient
temperatures could be an important factor enabling the spread of L.
braziliensis and L. infantum into new areas and the risk of spread of
both studied Leishmania species to higher latitudes and altitudes
should not be over looked. Another factor affecting Leishmania
development in vectors is the blood meal digestion by midgut
proteases. Previously, some studies have demonstrated that avian
blood inhibits Leishmania infection in the Old World vectors.
However, this effect has never been observed in the New World sand
flies. Therefore, our study was focused on the effect of chicken vs.
rabbit blood on sand fly digestion and the development of Leishmania
major in its natural vector P. duboscqi. In addition, we tested the effect
of avian blood on midgut trypsin activity and the influence of repeated
feedings on the susceptibility of sand flies to Leishmania infection.
Sand fly females fed on chicken blood had significantly lower midgut
trypsin activity and delayed egg development compared to those fed
on rabbits. On the other hand, there was no effect of avian blood on
parasite development within the sand fly gut. Similarly, blood feeding
of sand flies on birds or mammals prior to the infectious meal did not
cause any differences in Leishmania infections. Our study proved that
The goal of the Sand Fly Genome Sequencing Project was to
sequence the genomes of two different phlebotomine sand flies,
Phlebotomus papatasi and Lutzomyia longipalpis, that exhibit
distinct distributions, behavior, and pathogen specificity. Funded by
United States National Human Genome Research Institute the Sand
Fly Genome Project was initiated by a proposal submitted by Drs.
Mary Ann McDowell, Frank Collins, Marcelo Ramalho-Ortigao,
Jesus Valenzuela, Shaden Kamhawi, Rod Dillon, Paul Bates, and
Michael Lehane. The Lu. longipalpis portion of the project is headed
by Dr. Rod Dillon, Lancaster University, with sequencing at Baylor
College of Medicine Sequencing Center, USA. The Ph. papatasi
portion of the project is headed by Dr. Mary Ann McDowell,
University of Notre Dame, USA with sequencing at Washington
University Sequencing Center, USA. The genome of Lu. longipalpis
(Jacobina strain) is approximately 150 Mb at 38.9x coverage. The
N50 of the Lu. longipalpis contigs is 7.5 kb and the N50 of the
scaffolds is 85.1 kb. The genome of Ph. papatasi (Israeli strain) was
sequenced at 22.5x coverage and is approximately 350 Mb with a
scaffold N50 of 23.6 kb. Genome assemblies of both species are
available on VectorBase (https://www.vectorbase.org). Other
genomic resources available from this project include a 40,000 ESTs
from a normalized cDNA library of Ph. papatasi generated from the
four larval stages, pupa, male and female at 1, 3, and 10 days post
emergence and females post feeding on uninfected and Leishmania
major infected mouse blood. RNAseq data is also available for these
same conditions. In addition, the transcriptomes of Lu. longipalpis
larval stages and females fed with sugar and uninfected or infected
(Le. infantum) blood meals were sequenced. Genomic DNA from
males and females from both species was used to generate individual
BAC libraries. Further, SNP discovery is ongoing from individual
flies collected from different geographical areas. Manual annotation
is ongoing, but nearing completion. The following gene groups are
being assessed: genes involved in digestion and immunity; circadian
rhythm; microRNAs; G-protein coupled receptors, odorant and
gustatory receptors; salivary protein genes, heat shock protein genes,
cuticular protein genes, hormonal signaling, antioxidants,
aquaporins, genes involved in symbiotic interactions, transposable
N° 26 (1) 2015
elements, and potential viral sequences.
info@vectorbase.org.
THE DATABASE VECTORBASE AND THE GENOMES
OF LUTZOMYIA LONGIPALPIS AND PHLEBOTOMUS
PAPATASI
MANUAL ANNOTATION OF G PROTEIN–COUPLED
RECEPTORS (GPCRs) IN THE GENOMES OF
LUTZOMYIA LONGIPALPIS AND PHLEBOTOMUS
PAPATASI
Gloria I Giraldo-Calderón, Scott J. Emrich, Daniel Lawson, Frank H
Collins and the members of the VectorBase consortium
53
Samuel Tadros, Mariha Wadsworth, Mallory Hawksworth, RJ Nowling,
Mary Ann McDowell, Gloria I Giraldo-Calderón
University of Notre Dame, European Molecular Biology Laboratory, EMBL - European
Bioinformatics Institute, EBI, Institute of Molecular Biology and Biotechnology, IMBB,
Harvard University, Imperial College London, The University of New Mexico
Department of Biological Sciences, Eck Institute for Global Health, University of
Notre Dame, Notre Dame, IN 46556, USA.
ggiraldo@nd.edu
ggiraldo@nd.edu
Keywords: genomics, bioinformatics, database, genome
sequencing
Keywords: G protein–coupled receptors (GPCRs), gene manual
annotation, new insecticide targets
VectorBase (www.vectorbase.org) is a Bioinformatics Resource
Center, which hosts the genomes of invertebrate vectors of human
parasites and pathogens, including mosquitoes, tsetse flies, ticks, lice,
kissing bugs and two sandflies, Lutzomyia longipalpis and
Phlebotomus papatasi. Scientists working with sandflies in the field
and/or the laboratory can become VectorBase users. The data
deposited in VectorBase and in the public repositories, (such as
GenBank/NCBI, EMBL/EBI or the DDBJ), are constantly updated
and are a resource that has been subject to only very limited analysis to
date. These data are freely available for new analyses, descriptions and
hypothesis testing. Both public and private data can be queried or
analyzed with a variety of available tools including VectorBase Blast,
ClustalW, Hmmer, BioMart, Galaxy, Web Apollo and the Genome,
Ontology, Expression and Population Biology browsers. The
overarching goal of VectorBase is to support efforts to improve or
develop new vector control or eradication strategies. Questions about
how to use VectorBase, or suggestions about how to improve this
resource, are very welcome, please send your comments to
Sequencing the genomes of Phlebotomus papatasi and Lutzomyia
longipalpis will help in the task to combat sand fly-borne diseases
such as leishmanaisis. These sequences will provide valuable
resources for improving control or eradication strategies to stop the
transmission of these diseases. One such strategy is the use of G
protein-coupled receptors (GPCRs) as new targets for insecticide
chemistries. GPCRs play roles in many essential insect pathways and
can be specifically targeted with small molecules, making GPCRs
good insecticide targets. We used bioinformatic approaches to
identify GPCRs in the genomes of these two sand flies. A new
classifier pipeline called Ensemble*, was created to identify
candidate GPCR peptides from the P. papatasi and L. longipalpis
genome at Vector Base using the set of known GPCR peptides from
Aedes aegypti, Anopheles gambiae, Apis mellifera, Drosophila
melanogaster, Homo sapiens, and Pediculus humanus. The identified
sand fly GPCR sequences were aligned using BLAST against the
non-redundant protein sequences (nr) in NCBI. Putative functions
were assigned based on the best and the most informative BLAST
hits, in most of the cases the latter was from D. melanogaster. The
sand fly sequences were aligned with homologous genes, mostly
from insects used in the classifier pipeline, using MultAlin. The sand
fly gene models were manually corrected using the annotation tool
Artemis. Identified and manually annotated GPCRs were used to
search (tBLASTn) the sand flies genome scaffolds for additional
GPCR genes. When available, the gene models were confirmed using
transcript evidence from Expressed Sequence Tags (ESTs) and RNA
sequencing (RNAseq). The genes were named according to
nomenclature guidelines developed for invertebrate vectors as
detailed at VectorBase. If in doubt about the putative function of a
gene, a literature search and a tree reconstruction was done using the
software Clustal Omega. The same GPCR symbols used for other
VectorBase genomes were also adopted, to facilitate the database
query. The annotated gene models were submitted to VectorBase.
Examples of conservation between homologous genes, lineagespecific gene duplications/deletions and the fragmented nature of the
sand fly genome assemblies are also discussed.
N° 26 (1) 2015
54
GENETIC VARIATION OF NYSSOMYIA TRAPIDOI
(DIPTERA: PSYCHODIDAE) IN THREE ENDEMIC
ZONES OF LEISHMANIASIS IN THE ECUADOR
G E N E T I C A S S E S S M E N T O F S A N D F LY
POPULATIONS IN PANAMA
Gabriela Vela, Gabriel Trueba, Jérôme Depaquit, Sonia Zapata
Anayansi Valderrama Cumbrera, Mara Garcia Tavares, Ingrid Murgas,
José Andrade Dilermando
Universidad San Francisco de Quito, Ecuador; Université de Reims Champagne
Ardennes, France
Instituto Conmemorativo Gorgas de Estudios de la Salud (ICGES), Universidade
Federal de Viçosa, Centro de Pesquisas René Rachou
szapata@usfq.edu.ec
avalderrama@gorgas.gob.pa
Keywords: Ny. trapidoi, DNAmt, sympatry
Keywords: Sandfly, cutaneous leishmaniasis, genetic population,
genetic marker
In the present study entomological collections were carried out
between February and April 2013 in 3 provinces of Ecuador (Bolívar,
Esmeraldas y Manabí). There were 14 species of sand flies identified,
and Pressatia dysponeta, Psychodopygus panamensis Lutzomyia
hartmanni and Nyssomyia trapidoi were the more abundant. Ny.
trapidoi was found in 6 out of 7 points collected. The nucleotide
analysis of the mitochondrial genes, cytochrome oxidase subunit I
(COI) and cytochrome B (CytB) of 45 studied specimens allowed us
to group this specimens in four clusters One of the clusters had
specimens from all the studied provinces; meanwhile the other three
had specimens captured only from Bolivar province. This results
suggest that Ny. trapidoi is going through sympatric (inside Bolivar)
and allopatric speciation (Between populations in Bolivar and the
other provinces). Finally, it was found that 12,2% of the female
samples analyzed were infected with Endotrypanum monterogeii.
Many approaches have been implemented to understand the role of
Phlebotominae sandfly in the dynamics transmission of
leishmaniasis parasites. However, the introduction of genetic marker
has allowed assessment of sandfly populations and the progress in the
epidemiology understanding of leishmaniasis disease. In Panama,
reports of human cutaneous leishmaniasis during the 60´s, 70´s and
80´s were attributed to sylvatic area and Lu. panamensis, Lu. trapidoi
and Lu. gomezi as potential vectors. Since then, there have been
modifications of the landscape, climate and an increase number of
human cases is registered. In order to re-evaluate some criteria of the
disease cycle aimed to study the genetic populations of Lu.
panamensis, Lu. trapidoi and Lu. gomezi, therefore we estimate the
genetic diversity, gene flow and demographic history of these species
to infer the ability of survival and adaptability due change
environmental. Individuals were collected with CDC light traps and
identified to species level with taxonomic keys. DNA extraction of
individuals were performed using phenol chloroform method and
PCR amplification were implemented using the mitochondrial genes
cytochrome oxidase I and the end region of the cytochrome b,
intergenic region IGS -1, Ser - tRNA, IGS -2 and the initial region of
the fragment NADH1 and intron elongation factor α-1. Preliminarily
our results suggest that Lu. gomezi species showed a high genetic
diversity and slight population structure may be due to the migration
and local adaptation to environmental modifications by
anthropogenic intervention. Knowledge of changes in the structure of
Lutzomia genetics population may be helpful to understand the
changes in the patterns of dynamics transmission of cutaneous
leishmaniasis in Panamá.
N° 26 (1) 2015
EVIDENCE FOR GENETIC DIFFERENTIATION AT
THE MICROGEOGRAPHIC SCALE IN
PHLEBOTOMUS PAPATASI POPULATIONS FROM
SUDAN
GENE EXPRESSION PROFILING AND SILENCING
OF KEY DIGESTIVE ENZYMES IN LUTZOMYIA
LONGIPALPIS
Noteila M Khalid, Marium A Aboud, Fathi M Alrabba, Dia-Eldin A
Elnaiem and Frederic Tripet
Department of Zoology, Khartoum College of Medical sciences, Khartoum, Sudan.
55
Hector M Diaz-Albiter, Caroline d Moraes, Maiara d Faria, Mauricio R
Sant'Anna, Roderick J Dillon and Fernando A. Genta
Fundação Instituto Oswaldo Cruz (FIOCRUZ), Brazil
hector.albiter@ioc.fiocruz.br
Noteila@hotmail.com
Keywords: digestion enzymes, RNAi, Lutzomyia
Keywords: Phlebotomus papatasi, Sudan, Gene flow, Genetic
differentiation.
Background: Cutaneous Leishmaniasis (CL) is endemic in Sudan. It is
caused by Leishmania major parasites and transmitted by
Phlebotomus papatasi sandflies. Recently, uncommon clinical
manifestations of CL have been reported. Moreover, L. donovani
parasites that cause Visceral Leishmaniasis (VL) have been isolated
from CL lesions of some patients who contracted the disease in
Khartoum State, Central Sudan with no history of travelling to VL
endemic sites on south-eastern Sudan. Because different clinical
manifestations and the parasite behaviour could be related to genetic
differentiation, or even sub-structuring within sandfly vector
populations, a population genetic study was conducted on P. papatasi
populations collected from different localities in Khartoum State
known for their uncommon CL cases and characterized by contrasting
environmental conditions. Methods: A set of seven microsatellite loci
was used to investigate the population structure of P. papatasi samples
collected from different localities in Khartoum State, Central Sudan.
Populations from Kassala State, Eastern Sudan and Egypt were also
included in the analyses as outgroups. The level of genetic diversity
and genetic differentiation among natural populations of P. papatasi
was determined using FST statistics and Bayesian assignments.
Results: Genetic analyses revealed significant genetic differentiation
(FST) between the Sudanese and the Egyptian populations. Within the
Sudanese P. papatasi populations, one population from Gerif West,
Khartoum State, exhibited significant genetic differentiation from all
other populations including those collected as near as 22 km.
Conclusion: The significant genetic differentiation of Gerif West P.
papatasi population from other Sudanese populations may have
important implication for the epidemiology of leishmaniasis in
Khartoum State and needs to be further investigated. Primarily, it
could be linked to the unique location of Gerif West which is confined
by the River Nile and its tributaries that may act as a natural barrier for
gene flow between this site and the other rural sites. The observed high
migration rates and lack of genetic differentiation among the other P.
papatasi populations could be attributed to the continuous human and
cattle movement between these localities.
Lutzomyia longipalpis is the most important vector of American
Visceral Leishmaniasis. Adults of both sexes feed on sugars, while
adult females are also haematophagous and larvae feed on solid
detritus. Information regarding digestion in sand fly larvae is scarce.
However, glycosidase activities involved in microorganism digestion
have been described. In this work, β-1,3-glucanases, chitinases and
digestive lysozymes were identified. Tissue-specific expression
profiling was performed and physiological effects of RNAi-mediated
silencing of these enzymes were explored. This is the first study to
identify the molecular nature of sand fly larval digestive enzymes as
well to assess their relevance by reverse genetics.
N° 26 (1) 2015
ID 68-O Virus Infections
ID 69-O Virus Infections
EVALUATION OF COLLECTION METHODS FOR
PHLEBOTOMUS-BORNE VIRUSES DETECTION:
I S O L AT I O N A N D V I R A L R N A I N T E G R I T Y
PERFORMANCE
CONDITIONED MEDIA AND EXOSOMES FROM
LUTZOMYIA LONGIPALPIS LL5 CELLS DSRNATRANSFECTED ARE ABLE TO INDUCE NONSPECIFIC ANTIVIRAL RESPONSE IN LL5 NAIVE
CELLS
Remoli ME1, Bongiorno G2, Fortuna C1, Marchi A1, Bianchi R2, Khoury
C2, Ciufolini MG1, Gramiccia M2
1
Unit of Viral diseases and attenuated vaccine and 2Unit of Vector-borne Diseases and
International Health, MIPI Department, Istituto Superiore di Sanità, Rome, Italy
marina.gramiccia@iss.it
Keywords: Phlebotomus-borne viruses, Phlebotomus perniciosus,
Toscana Phlebovirus, Experimental infection
56
Andrea Martins da Silva1, Antonio Jorge Tempone1, André N Pitaluga1,
Fabricio K Marchini2, Barbara Burleigh3, Yara M Traub-Cseko1
1
Laboratório de Biologia Molecular de Parasitas e Vetores Instituto Oswaldo Cruz FIOCRUZ Pavilhão Leonidas Deane, Sl. 304. Av. Brasil, 4365. Manguinhos. Rio de
Janeiro, RJ – Brasil, CEP 21040-360 Tel.: +55 21 3865-8206 / Fax: +55 21 25903495; 2Laboratório de Genômica Funcional Instituto Carlos Chagas / FIOCRUZ-PR
R. Prof. Algacyr Munhoz Mader, 3775 CIC Curitiba – PR – Brasil, 81350-010 Phone
+55 41 3316 3236 Fax +55 41 3316 3267; 3Department of Immunology and Infectious
Diseases 665 Huntington Avenue Building 1, Room 817 Boston, Massachusetts 02115
Phone: 617.432.2495.
andrea.silva@ioc.fiocruz.br
Phlebotomus-borne (PhB) viruses are RNA viruses involved in
human disease outbreaks. Because several emerging viruses have
been recently isolated from sand flies, an important task is to monitor
the spreading of PhB-viruses. In order to investigate on collection and
storage methods of phlebotomine vectors for reliable RNA detection
and virus isolation, experimental viral infections of colonized sand
flies were carried out and different laboratory conditions were tested.
Laboratory-reared Phlebotomus perniciosus was experimentally
infected with Toscana Phlebovirus (TOSV, ISS-Phl3 strain) using a
membrane feeding technique. After the infected blood meal, engorged
females were stored by two methods: 1) alive sand flies (AS) were
immediately frozen at –80°C, mimicking the golden standard CDC
light trap collection procedure; 2) insects were put onto sticky traps
left at room temperature, collected daily up 144 h and stored dry at 80°C. Batches were separated into dead sand flies contaminated by
alcohol necessary for removal from papers (DS-A) and those in which
contamination was avoided (DS). The specimens, collected
individually or in pools, were homogenized and used both for virus
isolation in VERO cells and genome identification by Reverse
Transcription-PCR on N gene region, and Real Time-PCR on TOSV L
region. TOSV isolation percentage and RNA detection was 100% for
the AS specimens, as expected. In DS specimens the TOSV isolation
on VERO cells was elevated at 24 h but less satisfactory thereafter,
suggesting that the virus could be fairly isolated from infected wild
specimens only if they were frozen 24 h from death. No TOSV
isolation was obtained from DS-A specimens at any collection time.
As regards RNA detection, for all conditions examined Real TimePCR gave positive results until to 6 days after the infection. Recent
investigations have indicated that virus diversity in the Mediterranean
basin is higher than initially suspected, therefore a standardization of
reliable sand fly collection and storage methods is important to
harmonize entomological surveys. We confirmed that live specimens
are the best target for both viral isolation and RNA identification.
However sticky traps are suitable for partial genome viral
identification but less so for viral isolation. This study was carried out
in the frame of EDENext, a collaborative project of the 7th FP (20112014) funded by the European Commission under the DG Health;
Contract Number: 261504.
Keywords: antiviral response, conditioned media, exosomes
Phlebotomine sand flies are haematophagous insect vectors of
leishmaniasis in the New and Old World. In the Old World these
insects are also related to virus and bacteria transmission. Although
there is no proven scientific data on the New World sand flies capacity
to transmit viruses, these insects may be considered as potential
disseminators of these pathogens. The Lutzomyia longipalpis is the
principal vector of visceral leishmaniasis in the Americas. Previous
results of our group identified a nonspecific antiviral response in
cultured L. longipalpis embryonic cells line LL5, in response to
transfection with double stranded RNA (dsRNA). This was the first
report of this kind of response in an insect cell line. We are presently
interested in identify the mechanisms by which LL5 cells recognize
dsRNA and trigger the antiviral response. Recently the role of
secreted molecules in the host cells and pathogens has increased
dramatically. In this work we incubate naive cells with conditioned
media or isolated exosomes from dsRNA transfected and mocktransfected (control) LL5 cells, then infected these treated cells with
West Nile virus-like particles expressing luciferase. Interestingly the
cells incubated with conditioned media or exosomes from dsRNAtransfected cells, displayed a statistically significant antivirus
response when compared with control groups. Because of this result,
we decided study the protein composition of each one of these
conditioned media (24 and 48 hours after transfection) and from
exosomes isolated from these media. Samples from both sources
were analyzed using mass spectrometry methodologies. Initial
analysis identified 306 peptides in the conditioned media and 530 in
exosomes. Some of these proteins were associated with antiviral
response and/or structural vesicles. Among these proteins we can cite
the Scramblase, FKBP, Adaptor protein and Nascent polypeptideassociated complex. These proteins were highly incident in
exoproteoma from cells 24 hours after transfection, however,
exhibited a reduction when analysed the exoproteome in 48 hours
after transfection when we compared with control.
The
transcriptional pattern of some of these proteins was investigated in
new LL5 cells cultures dsRNA-transfected. This is a first report on
the exoproteome analysis related to antiviral response in
phlebotomine embryonic cells.
N° 26 (1) 2015
ID 70-O Physiology & Colonization
CD-ROM SAND FLY FELLAS: THE FIRST COMPLETE
VIDEO-BASED SAND FLY REARING GUIDE IN HIGH
DEFINITION
COMPARISON OF VOLATILE COMPOUNDS
EMITTED BY ANIMALS OR PRESENT IN PLANTS ON
ATTRACTIVENESS OF NYSSOMYIA NEIVAI
(DIPTERA: PSYCHODIDAE)
Meneses C, Morales E, Rhodes C, Oliveira F, Valenzuela JG and
Kamhawi S.
NIAID, NIH, Rockville, Maryland, USA.
claudio.meneses@nih.gov
Keywords: Sand Fly, Video Based, Rearing guide, Leishmaniasis
It is well known that less than a hand full of sand fly species can be
reared in the laboratory in sufficient numbers to provide enough
material for large-scale experiments. Large-scale sand fly
colonization is challenging and requires knowledge of sand fly
biology, availability of the appropriate infrastructure and equipment,
and having access to up to date protocols. With the resurgence of
leishmaniasis, a neglected tropical disease, and the demonstration of
the virulence of vector-transmission compared to needle-delivered
parasites, significant investments have been made by institutions and
companies to evaluate new leishmaniasis vaccines using infected sand
flies. Development of experimental models of natural transmission
requires a transition from small sand fly colonies to large-scale
colonies. The objective of this presentation is to provide an account of
the steps required to grow a large-scale sand fly colony and how to
prevent or overcome problems that commonly arise during
colonization. Our sand fly insectary unit, a branch of the Laboratory of
Malaria and Vector Research (LMVR) at the National Institutes of
Health, has acquired expertise in mass rearing of two major sand fly
species: Lutzomyia longipalpis from Jacobina, Brazil and
Phlebotomus duboscqi from Mali, Africa providing an annual
productivity averaging 1.2 million reared sand flies and an
experimental consumption of about 120,000 flies. With the goal of
contributing to the sand fly community, we are sharing our expertise
with other insectaries and interested laboratories through a 2.8 GB
Windows© based CD-ROM with videos in high image definition
embedded on a friendly user auto run menu designer™. The CD-ROM
also contains sand fly rearing techniques according to our most up to
date protocols; a complete list of materials needed to rear sand flies
along with schematic drawings (AUTO CAD©) of custom-made
items indispensable for handling a large number of flies on a daily
basis with precision; a comprehensive list of references on sand fly
rearing with links to PubMed; a link to Bei Resources where
laboratories in need can request sand fly eggs or larvae of available
vector species to initiate their own colonies; and a link to our lab where
we can offer further advice on how to mass rear sand fly species of
interest.
57
Vicente Estevam Machado1; Thais Marchi Goulart2; Arlene Gonçalves
Corrêa3; Mara Cristina Pinto1
Faculdade de Ciências Farmacêuticas-UNESP (Brasil)1; Instituto de Biologia UNICAMP (Brasil)2; Departamento de Química -UFSCar (Brasil)3
vicentemachado@yahoo.com.br
Keywords: primary alcohols, kairomone, wind tunnel, attractiveness
Chemical ecology approach is a feasible tool for monitoring and
maybe with intensive studies controlling species of haematophagous
insects. Usually the studies are concentrated in the search for volatiles
emitted from hosts which attract haematophagous insects
(kairomones). Considering that males and females sand flies require
sugar meal to survive, plants compounds can also be an interesting
source for chemical ecology researches. In this study, we aimed to test
the sand flies attractiveness in wind tunnel using two kairomones
from hosts used in previous studies for haematophagous insects
(lactic acid and 1-octen-3-ol) and primary alcohols that are usually
found in many types of plants: 1- propanol, 1-butanol, 1-pentanol, 1hexanol, 1-heptanol, 1-octanol, 1-nonanol e 1-decanol.The tests in
the wind tunnel were carried out with Nyssomyia neivai, an important
vector of cutaneous leishmaniasis in South America. Groups of three
female sand flies were placed inside releasing chambers in a total of
30 insects for each test and positioned 50 cm far from the stimulus. At
the beginning we tested 200 µL in filter paper of each compound in
three different concentration 10%, 50%, 100% (neat), if necessary the
concentration of 5% was also included. Tests lasted two minutes and
activation (number of insects that leave the release chamber) and
attraction (number of insects that reach the stimulus) behaviors were
evaluated. For the control group we used the diluents: hexane for
alcohols and ethanol for lactic acid. Sand flies responded in a dose
dependent way to 1-butanol, 1-pentanol, 1-hexanol, 1-octanol and 1octen-3-ol. The best responses for both behaviors evaluated were
obtained with 1-hexanol, 1-octanol and 1-octen-3-ol. Lactic acid
presented a weak response of activation and no attraction response.
This result is in accordance with mosquitoes studies which have
shown an activation role for lactic acid. The results showed that
compounds from plants, like primary alcohols, may be an useful
source for searching new compounds for trapping sand flies.
N° 26 (1) 2015
ID 72-O Physiology and Colonization
RESPONSES OF THE SAND FLY PHLEBOTOMUS
PAPATASI SCOPOLI (DIPTERA: PSYCHODIDAE) TO
AN OVIPOSITION PHEROMONE ASSOCIATED WITH
CONSPECIFIC EGGS
STUDY ON THE BACTERIAL MIDGUT MICROBIOTA
ASSOCIATED TO IMMATURE STAGE OF GENUS
LUTZOMYIA (DIPTERA: PSYCHODIDAE) ISOLATED
FROM NATURAL BREEDING SITES.
Ifhem Chlebi1-2, Elyes Zhioua1, and Gordon Hamilton2
Vivero J Rafael1, 2; Cadavid Gloria3; Moreno H Claudia Ximena3;
Posada-López Laura 1, 2; Sandra Uribe S. 1, 2
1
Institut Pasteur de Tunis, Laboratory of Vector Ecology, Tunis, Tunisia; 2Centre for
Applied Entomology and Parasitology, Keele University, Stafordshire, UK
ifhemc2001@yahoo.fr
Keywords: Phlebotomus papatasi, oviposition pheromone, eggs.
The chemical ecology of Lutzomyia longipalpis the South American
vector of Leishmania chagasi infantum has been extensively studied
and this species complex is known to produce both sex and oviposition
pheromones. By contrast very few studies on the pheromones of Old
World sandflies have been performed. Our preliminary studies have
shown that female P. papatasi produce an oviposition pheromone.
Female Phlebotomus papatasi laid a significantly (T-test; n=12 ; P
<0.01) greater number of eggs on sites with 160 eggs already present
than on control sites with no eggs. When smaller numbers of eggs (40
or 80) were used no significant differences between the mean numbers
of eggs laid on test and control sites were detected. Furthermore when
extract was prepared in hexane from batches of 160 eggs and placed on
filter paper in test pots a significant increase in the number of eggs laid
compared to the controls was found (Paired T-test, n=8, P < 0.05).
Together these results indicate the presence of an oviposition
pheromone on the eggs of P . papatasi.
58
1
Laboratorio de Sistemática y Biología de Insectos (Insectario), Grupo de
investigación en Sistemática Molecular, Facultad de Ciencias, Universidad Nacional
de Colombia, sede Medellín; 2PECET (Programa de Estudio y Control de
Enfermedades Tropicales Unidad de Entomología Médica y Molecular, Universidad
de Antioquia; 3Grupo de Microbiodiversidad y Bioprospección. Universidad Nacional
de Colombia-Sede Medellín
rjviverog@unal.edu.co
Keywords: immature, digestive tract, bacterial communities
Background: Few researches provide information about the bacterial
communities associated with the digestive tract of genus Lutzomyia
and their possible role in the dynamics of transmission of the
Leishmaniasis. The development of immature stages of genus
Lutzomyia can include great variety of sites that contain decomposing
organic material, where larvae may acquire different bacterial
communities. The identification of such bacteria is of importance
because their potential uses in designing improved vector control
strategies. Bacterial communities could regulate the reproduction,
abundance, distribution of Phlebotomine and can help identify the
breeding sites of genus Lutzomyia. This premise has motivated new
initiatives directed to study the bacterial communities associated with
the digestive tract of immature individuals of genus Lutzomyia (found
in natural breeding sites). Materials: Wild populations of immature
stages of genus Lutzomyia, were collected in Ovejas (department of
Sucre) in the Caribbean region of Colombia. The digestive tract of
larvae and pupae were dissected. Digestive tracts were spread in a
simple broth for enrichment. Genomic DNA was isolated from each
of the pure cultures and amplified (PCR-Colony) the intergenic
region of the 16S-23S rDNA and of the 16S gen which was sequenced
to each isolate. The resulting sequences were compared with the
GenBank database using the BLAST algorithm and software RDP.
Phylogenetic affiliations of culturable bacteria and a NeighborJoining (NJ) dendrogram were constructed and the relevance of
bacteria isolated from the gut was observed. Results: Our results
indicated that there is a dominance of gram-positive bacteria in the
digestive tract and the Malpighian tubules of immature individuals of
genus Lutzomyia. Different ITS profiles were observed among
immature stages (larvae and pupae). These results revealed a different
flora in immatures of genus Lutzomyia (Streptomyces cinnabarinus,
Ochrobactrum anthropi, Shinella zoogloeoides, Microbacterium
pseudoresistens, Bacillus cereus, Paenibacillus illinoisensis,
Lysobacter soli), when compared with other studies that only
contemplate the adult stage of phlebotomine sand flies. Conclusion:
This work provided basic information about bacterial diversity in
midgut of immature stages of genus Lutzomyia. Some bacterial
species are ideal targets that could be used for control.
ABSTRACT BOOK
POSTERS
ISOPS VIII
Puerto Iguazú
Argentina
N° 26 (1) 2015
ID 1-P Control
ID 2-P Control
SANDFLY FAUNA (DIPTERA: PSYCHODIDAE) OF
URBAN AREA IN THE CITY OF
SANDFLY FAUNA (DIPTERA: PSYCHODIDAE) FROM
REGIONAL OF HEALTH IN
SINOP, MATO GROSSO, BRAZIL
SINOP, MATO GROSSO, BRASIL, YEAR 2013
Sirlei Franck Thies1,2,3, Ana Lucia Maria Ribeiro3,5, Jorge Senatore
Rodrigues Vargas3,5, Edelberto Santos Dias4, Amilcar Sabino Damazo1
Sirlei Franck Thies1, 2, 3, Marinês Browers2
1
Laboratory of Histology, Department of Basic Sciences in Health, Medicine College,
UFMT, Cuiabá/MT. 2Regional Office of Health of Sinop, State Department of Health of
Mato Grosso, Sinop/MT. 3Laboratory of Medical Entomology, Medicine College,
UFMT, Cuiabá/MT. 4Laboratory of Leishmaniasis, Center of Research René
Rachou/Fiocruz-Minas Gerais. 5University Hospital Júlio Müller, UFMT, Cuiabá/MT.
60
1
UFMT, Cuiabá/MT.
sfthies@hotmail.com
sfthies@hotmail.com
Keywords: Leishmaniasis, Sandfly, Lutzomyia
Keywords: Leishmaniasis, Sandfly, Lutzomyia
Mato Grosso State registers cases of autochthony of American
Cutaneous Leishmaniasis (ACL) in 100% of its cities, and Sinop is
one of the three cities in spotlight in number of cases. For Visceral
Leishmaniasis (VL), a growing number of cities are registered with
reports of autochthonal cases for this harm, a fact that worries health
authorities, since they registered a human case in 2010 in Sinop. Sinop
is located in the northern of Mato Grosso and has been a spot on the
national stage for its socioeconomic growth, making it a commercial,
educational and health polo. This study aimed to diagnose the sandfly
fauna in the urban area in the city of Sinop, Mato Grosso.
Entomological collections were made using CDC light traps, which
were installed at 6 p.m. and collected at 7 a.m. the next day, during
three nights in a row, in November 2013, around the homes located in
24 neighborhoods in the urban area of Sinop. The captured insects
were prepared and identified in the Laboratory of Entomology of the
State Department of Health of Mato Grosso. Thirty-three (33)
specimen of sandflies, 17 male (51.55%) and 16 (48.5%) female, were
captured. The captured sandflies belonged to 13 different species. The
main species was Lutzomyia davisi (21.2%), followed by Lutzomyia
antunesi (18.2%). Half of the neighborhoods were positive for
sandflies. Other important species in the epidemiology of ACL were
diagnosed, but at lower frequencies, such as: Lutzomyia whitmani
(main vector for Mato Grosso) and Lutzomyia yuilli yuilli. The
diagnosis of Lutzomyia whitmani, main vector of ACL in the state of
Mato Grosso and Lutzomyia antunesi, suspected species for
transmitting ACL and Visceral Leishmaniasis (VL), already found
naturally infected by Leishmania chagasi in Mato Grosso, highlights
the risk and the possibility of autochthonal transmission of ACL and
VL in the city, even in urban areas. We emphasize the need for
entomological and epidemiological monitoring, as well as the study of
natural infection of these insects by Leishmania, studies which also
involve the canine seroprevalence and the diagnosis of Leishmania
circulation in humans, so that actions of prevention, promotion and/or
control toward to the harm can be triggered, in order to minimize the
contact of man/vector and the reduction in the number of cases in the
city.
The Leishmaniasis are zoonotic diseases caused by protozoa of the
order Kinetoplastida, family Tripanostomatidae, genus Leishmania,
transmitted by insects of the genus Lutzomyia, called sandflies,
popularly known as white wing, stiff wing, birigui or straw
mosquitoes. This study aimed to diagnose the current sandfly fauna in
cities with reported cases of American Cutaneous Leishmaniasis
(ACL) in Regional of Health in Sinop, Mato Grosso, during the year
2013. To this, entomological surveys were conducted from July to
October 2013, with assistance from CDC light traps, installed at 6:00
p.m. and collected at 7:00 a.m. the following day, for three nights in a
row. Insects captured were sent to the Laboratory of Entomology of
the State Department of Health of Mato Grosso, where they were
prepared and identified according to Young and Duncan, 1994. The
Regional of Health in Sinop has 14 cities in its coverage area, making
it possible to have the research done in only 5 cities in the period. A
number of 3.029 sandflies were captured; the city with the largest
number of insects captured was Nova Ubiratã 2.618 (86.43%). There
was a predominance of male insects 1.985 (65.53 %). The city with
the greatest diversity of caught species was Nova Ubiratã with 23
distinct species. The most prevalent species was Lutzomyia lenti
1.208 (40.77 %). In all cities species of importance in the
epidemiology of ACL were diagnosed as: L. flaviscutellata and L.
whitmani (main vector in Mato Grosso), as well as in Nova Ubiratã it
was diagnosed L. longipalpis, vector of Visceral Leishmaniasis (VL).
The diagnosis of the presence of vector species of ACL and VL lead us
to recommend preventative and protective measures ranging from
environmental management (sanitation) to activities of promotion
and protection of health that should be developed along with the
community, keeping a focus at minimizing human contact with the
vector.
N° 26 (1) 2015
ID 3-P Control
ID 4-P Control
TESTING THE VERSATILITY OF A PSYCHODIELLA
SP. DIAGNOSTIC ASSAY IN
KINETICS OF ANTI-PHLEBOTOMUS PERNICIOSUS
SALIVA ANTIBODIES IN MICE
FIELD SURVEYS
AND RABBIT SERA
Lorena G. Caligiuri1,2, Enrique A. Sandoval3, María Soledad Santini4,
Soraya Acardi5, Oscar
Inés Martín-Martín, Ricardo Molina, Maribel Jiménez
D. Salomón6, Lilian Tartaglino3 and Christina B. McCarthy1,2
61
Unidad de Entomología Médica, Servicio de Parasitología, Centro Nacional de
Microbiología, Instituto de Salud Carlos III, Ctra. Majadahonda-Pozuelo s/n, 28220,
Majadahonda, Madrid, Spain.
1
Centro Regional de Estudios Genómicos, Facultad de Ciencias Exactas, Universidad
Nacional de La Plata, La Plata, Argentina.2 Departamento de Informática y Tecnología,
Universidad Nacional del Noroeste de la Provincia de Buenos Aires, Pergamino,
Buenos Aires, Argentina.3 Laboratorio de Vectores, Secretaría de Calidad de Vida,
Municipalidad de Posadas, Posadas, Misiones, Argentina.4 Laboratorio de Biología
Molecular Aplicada, Facultad de Ciencias Exactas, Químicas y Naturales,
Universidad Nacional de Misiones, Posadas, Misiones, Argentina.5 Centro Nacional de
Diagnóstico e Investigación en Endemoepidemias, Administración Nacional de
Laboratorios e Institutos de Salud, Ministerio de Salud, Buenos Aires, Argentina.6
Instituto Nacional de Medicina Tropical, Ministerio de Salud de la Nación, Puerto
Iguazú, Misiones, Argentina.
mccarthychristina@gmail.com
Keywords: Psychodiella chagasi, diagnostic assay, PCR, field
survey
Psychodiella chagasi natural infections have previously been
recorded in Lutzomyia longipalpis from Lapinha (Minas Gerais)
(Adler and Mayrink, 1961), Jacobina (Bahia) in Brazil (Lantova et al.,
2010) and, recently, Posadas (Argentina) (McCarthy et al. 2011).
Given the fact that the Argentine Lu. longipalpis population is
significantly differentiated from Brazilian populations (Salomon et al.
2010), we were interested in developing a straightforward and
sensitive assay to analyse the incidence of natural gregarine infections
in Lu. longipalpis from this location. In this context, we recently
reported the first PCR-based assay for the detection and identification
of natural gregarine infections in Lu. longipalpis (Caligiuri et al.,
2014). Briefly, sand fly gregarine diagnostic primers PsyF and PsyR
were designed on the basis of Ps. chagasi SSU rDNA sequences we
previously identified in male Lu. longipalpis from Posadas
(McCarthy et al. 2011). The specificity and sensitivity of the
diagnostic primers were confirmed by in silico analysis and in vitro
and field validations using total DNA extracted from naturally
infected Lu. longipalpis (Caligiuri et al., 2014). Having validated our
diagnostic assay for sand fly gregarine parasites in Lu. longipalpis, we
were interested in testing its versatility for subsequent use in field
surveys. Therefore, to evaluate the perfomance of the assay in a wider
range of field captured Lu. longipalpis, we analysed 159 captured
individuals from this species from Posadas (Argentina), using
samples made up of one sand fly each and pools of 5 and 10 sand flies
using different protocols and conditions. As expected, to obtain
consistent results for the samples made up of one sand fly each, it was
necessary to optimise the DNA extraction protocol. Our results
indicated that the diagnostic assay is very versatile for field surveys
and can be adapted to different sampling designs. Furthermore, in
accordance with our previous results, the diagnostic assay detected
gregarine infections that were not observed previously by light
microscopy analysis.
mjimenez@isciii.es
Keywords: Phlebotomus perniciosus, anti-saliva antibodies,
recombinant salivary proteins.
Anti-saliva antibody response can be used as marker of exposure to
sand flies. When hosts are bitten they develop humoral responses
against sand fly saliva. Moreover, a positive correlation has been
observed between number of bites and antibody levels. Little is
known about kinetics of antibodies against salivary gland
homogenate (SGH) or recombinant salivary proteins (rSP). This
work focuses on the study of anti-Phlebotomus perniciosus saliva
antibodies in sera of mice and rabbits that were experimentally
exposed to the bites of uninfected P. perniciosus. In addition, IgG
antibody levels against two P. perniciosus rSP, apyrase rSP01B
(GenBank ID: KF178455) and D7 related protein rSP04 (GenBank
ID: KF178456) were determined. Eight mice were immunized
weekly through the bite of 150 sand flies for 12 weeks and their
humoral response was followed by another 12 weeks. Six rabbits
were weekly exposed to 500 sand fly bites for at least 4 weeks and
their humoral responses were followed by 7 months. After nonexposure period both animals were exposed to test the presence of a B
cell memory recall. All animals were bled according to the
immunization schedule and anti-saliva antibodies were evaluated by
ELISA. Anti-saliva response was also tested in rabbits by Western
blot. In mice and rabbit sera, anti-saliva antibody levels increased
along the immunizations and correlated with the number of sand fly
bites. In both cases a memory response was highlighted. In mice,
anti-SGH antibody levels were detected in sera 4 weeks after
exposure, and persisted at least 3 months after the last exposure. Antiapyrase rSP01B antibodies followed similar kinetic responses that
SGH, but with detectable antibody levels at fifth week. Anti-D7
protein rSP04 antibody response showed high variability among sera
of immunized mice. In the case of rabbits, anti-saliva antibodies
appeared after the 2nd week of exposure and antibodies were
detected at 7 months post-exposure. In conclusion this study shows
valuable data on sand fly saliva response in mice and rabbits. Results
on kinetics of anti-saliva antibodies in rabbit sera provide useful
information for an accurate application of anti-saliva response as
marker of exposure in the field as wild rabbits have been proved to
transmit Leishmania infantum to P. perniciosus in the focus of human
leishmaniasis in south western Madrid region, Spain. Study
supported by the Spanish Project AGL2008-01592 and by EU grant
FP7-2011-261504 EDENext.
N° 26 (1) 2015
ID 5-P Control
ID 6-P Control
FIELD EVALUATION OF AN INSECTICIDE
FORMULATED WITH ACTIVE INGREDIENTS
ACTING ON LARVAL AND ADULT STAGES OF
LUTZOMYIA LONGIPALPIS
CHARACTERIZATION OF SUSCEPTIBILITY OF
POPULATIONS OF LUTZOMYIA LONGIPALPIS
( D I P T E R A : P S Y C H O D I D A E ) TO
ALPHACIPERMETRIN
Laura W. Juan1, 2, Ignacio T. Gould3, Oscar D. Salomón4, Juan Molina4,
Raul A. Alzogaray1, Eduardo N. Zerba1
Douglas de Almeida Rocha, Thaís Oliveira Coelho, Rodrigo Gurgel
Gonçalves, Andrey José de Andrade & Marcos Takashi Obara
1
Núcleo de Medicina Tropical, Universidade de Brasília, Campus Universitário Darcy
Ribeiro, Asa Norte, 70910-900, Distrito Federal, Brasil
CIPEIN–Centro de Investigaciones de Plagas e Insecticidas (UNIDEF-CONICET),
Villa Martelli, Buenos Aires. Argentina. 2Cátedra de Zoología. Facultad de Ciencias
Agrarias. UNLZ, Buenos Aires. Argentina.3CeNDIE –Centro Nacional de Diagnóstico
e Investigación en Endemoepidemias. Buenos Aires.REDILA 4 INMeT –Instituto
Nacional de Medicina Tropical, Puerto Iguazú, Misiones. Argentina.REDILA
ljuan@citedef.gob.ar
Keywords: Lutzomyia, chemical control, permethrin, pyriproxyfen
In Argentina, the American tegumentary leishmaniasis (ATL) is
endemic, with wild enzootic cycles, while visceral leishmaniasis (VL)
is urban. VL is more severe because it can affect internal organs with
high fatality. Since 2006, this disease is an emerging phenomenon of
public health importance. The most practical way of prevention is to
control the insect vector, phlebotomines sand flies from genus
Lutzomyia, by application of residual insecticides, and dogs treatment
with veterinary formulations. Recently, insecticide treated nets were
introduced with controversial results. The application of insecticides
in phlebotomines cities foci could be an important vector control tactic
to reduce urban transmission. With the objective to develop a new
control alternative, we evaluated a formulation containing the
pyrethroid insecticide permethrin (10%) and the insect growth
regulator pyriproxyfen (2%). According studies performed at our
laboratory, permethrin is effective on adult L. longipalpis.
Pyriproxyfen is an larvicide that disrupts metamorphosis of immature
stages of sand fly, preventing it from reaching the adult stage. The trial
was carried out in the city of Puerto Iguazú, Misiones, Argentina in
March-April, 2014. Three dwellings with presence of L. longipalpis,
all of them with poultry houses, were selected. Before applying the
formulation, population level of sand fly were monitored for two days
with CDC type traps. The formulation was emulsified in water and
applied in two dwellings (TP and Chan). The third house (Ct) was
untreated (control). The application was performed during sunset
using a hand pulverizer. Application covered the interior of houses and
surroundings to a distance of three meters. One week before the
application the average of number of adults sand flies catched by the
traps were 13 and 22 in the houses TP and Chan, respectively. Just
before the treatment the flies catched during two days were 21 and 11,
respectively. One month after the treatment, no flies were catched in
TP and just one in Chan. In the control house, more than 50 sandflies
were catched in all cases. This succesfull result could be attributed to
the complementary effects of the insecticides applied: permethrin
against adults insects and pyriproxyfen against larvae present in soil.
62
dougalmeidarocha@gmail.com
Keywords: Sand flies, control, insecticides, visceral leishmaniasis
Introduction: Visceral Leishmaniasis (VL) is an important public
health problem in many countries of Latin America. Nowadays, none
safe and effective vaccine exists and the transmission control of VL
depends combating insect vectors using application of residual
insecticides. Currently, there are records of outbreaks of sand fly
populations resistant to DDT and BHC, in Sri Lanka, India and
Venezuela. In Brazil, information about resistance of populations of
Lu. longipalpis are limited and little is known about the genetic
potential of this important resistance transmitter LV kind of man.
Objective: Characterize the susceptibility profile of populations of
Lu. longipalpis alfacipermetrina the pyrethroid used in Vector
Control Program in Brazil. Methods: Dose-response bioassays with
laboratory specimens (control) will be carried out by comparison
them field populations from four endemic for VL in Brazilian states.
Ten specimens of Lu. longipalpis will be exposed in bottles
impregnated with insecticide in four alphacipermetrin concentrations
to estimate the curve versus dose of insecticide mortality of insects. A
total of 80 adult of sand flies (40 males and 40 females), F1
generation, between three to five days of life will be divided into four
vials tests and one control. Reason of Resistance and Slope will be
calculated for all populations. It will be considered from 98 to 100%
mortality; as indication of susceptibility from 80 to 97%strength
confirmation; and less than 80% mortality resistance will point
established. All tests shall be repeated at least three times for each
population on different days. Final considerations: The results will
allow for the rapid and early detection of populations with probable
evidence of insecticide resistance, contributing to the “Programa de
Controle de Leishmaniose Visceral (PCLV)” of the Ministry of
Health of Brazil in the deployment of alternative vector control
strategies, as well as the periodic monitoring and systematic different
populations of Lu. longipalpis, coming from different areas of the
country.
N° 26 (1) 2015
63
ID 7-P Control
ID 8-P Control
EFFECTS OF LEISHMANIA MAJOR INFECTION ON
SAND FLY FEEDING RESPONSES TO DEET AND OR
PERMETHRIN TREATED UNIFORM MATERIAL
SOIL ANALYSIS AND VEGETATION ASSOCIATED
WITH THE BREEDING SITES OF LUTZOMYIA
EVANSI IN THE MUNICIPALITY OF OVEJAS-SUCRE
A FOCUS OF LEISHMANIASIS IN COLOMBIA
Tobin Rowland, Silas Davidson, Kevin Kobylinski, Edgar Rowton
Walter Reed Army Institute of Research, Silver Spring, MD, USA
tobin.e.rowland.civ@mail.mil
Horacio Cadena1, Edgar Ortega2, Luz A. Acosta1, Andrés Vélez-Mira1,
Rafael J. Vivero1, Eduar E. Bejarano2, Sandra Uribe3, Hernando
Gómez4, Luis G. Estrada2 & Iván D. Vélez1
1
Keywords: DEET, Permethrin, Leishmania major, Phlebotomus
duboscqi
Despite the use of Personal Protective Measures (PPMs) and other
vector control applications, cases of tropical diseases like malaria,
leishmaniasis, dengue, and other viral diseases still occur in deployed
military service members. Some of these pathogens may affect the
feeding behavior of the insects they infect. In sand flies, Leishmania
parasites attach to the stomodeal valve of the anterior midgut. This
attachment often interferes with the sand fly's ability to obtain a blood
meal which can result in multiple probes. The objective of this study
was to determine if Phlebotomus duboscqi infected with Leishmania
major parasites responded differently than uninfected sand flies to
commonly used PPMs; DEET applied to the skin and permethrin
treated uniform material. A hairless guinea pig model was used to
determine differences in the blood feeding rate and probing activity.
Twenty sand flies were allowed to feed in a modified cage placed on
anesthetized guinea pigs that had been treated with each PPM. DEET
was applied to the skin at the rate recommended for people and
permethrin treated uniform fabric was placed on the guinea pigs with a
8.75cm2 hole exposed. The Fisher's Exact test was used to compare
the proportions of blood fed sand flies within and between treatment
groups. Both the DEET and permethrin treated uniforms decreased
the ability of sand flies to successfully take a blood meal, and there was
no significant difference between infected and uninfected flies. When
permethrin treated uniforms were used without DEET, there was a
significance increase in the amount of probing activity by both
infected and uninfected flies. This suggests that permethrin treated
clothing may force sand flies to exposed skin and may actually
increase the risk of disease transmission if not used with a topical skin
repellent.
PECET (Programa de Estudio y Control de Enfermedades Tropicales), Universidad
2
de Antioquia, Medellín, Colombia.
Grupo de Investigaciones Biomédicas,
Universidad de Sucre, Sincelejo, Colombia. 3Grupo de Investigación en Sistemática
Molecular, Universidad Nacional, Medellín, Colombia. 4Atención Integral en Salud
(ATIS)- Corposucre
horaciocadena@gmail.com
Keywords: Leishmaniasis, Lutzomyia, breeding site, vegetation
Several studies have shown that the physical and chemical
characteristics of soil and vegetation are essential factors that
determine the breeding sites of sand flies. This study was conducted
in the peri-urban area of the municipality of Ovejas-Sucre, Colombia
a endemic focus of cutaneous and visceral leishmaniasis. This region
has two contrasting seasons: a rainy period from May to November
and a dry period from December to April. The annual precipitation
and mean temperature is 1380 mm and 28°C, respectively, and
according to Holdridge the ecological life-zone is Tropical Dry
Forest. The aim of this study was to describe the composition of soil
and vegetation associated with the breeding sites of immature forms
of Lutzomyia (Lu.) evansi the main vector of cutaneous and visceral
leishmaniasis in the Colombian Caribbean region. Soil samples
(n=43) were collected as follows: zone 1 (n=13), zone 2 (n=14) and
zone 3 (n=16). In the three zones evaluated an average pH value of
7.43 (range 7.10-7.68) indicated a marginally basic soil.
Exchangeable aluminum (Cmol. kg-1 soil) and salinity was zero. The
texture of the samples was classified as sandy loam and silt loam
soils. The organic content of the soils, which is important for larval
development, was ̅x 3.35% (range 0.76 -5.83) being on average
higher in the zone 1. In the three zones studied a total of 27 plots of
100 m2 each were sampled to characterize the vegetation associated
with positive breeding sites. The vegetation in zone 1and 2 was
characterized by herbaceous and shrub species with scattered trees. In
contrast, in zone 3 a greater number of trees were recorded. Larvae
(n=98 Lu. evansi) were predominantly recovered from the trees
Cordia dentata, Guazuma ulmifolia and Manguifera indica. These
results suggest that the knowledge of the peri and extradomiciliary
breeding sites of sand flies could help orient environmental
intervention measures that would affect the breeding sites of
sandflies.
N° 26 (1) 2015
ID 9-P Control
ID 10-P Control
THE USE OF SYSTEMIC INSECTICIDES TO
CONTROL PHLEBOTOMUS SPP. SAND FLIES IN
SOUTHERN KAZAKHSTAN
TRANSMISSION OF LEISHMANIA INFANTUM TO
LUTZOMYIA LONGIPALPIS BY HUMANS WITH AND
WITHOUT HIV INFECTION, SYMPTOMATIC AND
ASYMPTOMATIC
David Poché¹, Alex Gendernalik¹, Larisa Polyakova¹, Richard Poché¹,
Madina Kozhahmetova², Batirbek Aimakhanov², Ziyat Abdeliyev²,
Bakhit Atshabar²
¹Genesis Laboratories, Inc.: Colorado, United States. ²Kazakh Science Center for
Quarantine and Zoonotic Diseases by State Committee of National Sanitary-andEpidemiological Oversight of the Ministry of Health by Masgut Aikimbaev (KSCQZD):
Almaty, Kazakhstan
64
Carlos C B Ribeiro, Antônio Menezes Filho, Daniela M Parente,
Teresinha C Farias, Danielle A Zacarias, Jalthon C da Silva, Dorcas L.
Costa, Ivete L Mendonça, Carlos H N Costa
Universidade Federal do Piauí.
chncosta@gmail.com
davidp@genesislabs.com
Keywords: Leishmania infantum, Lutzomyia longipalpis,
reservoirs, xenodiagnosis
Keywords: Rhombomys opimus, Paraphlebotomus mongoliensis,
fipronil, Bakanas
Cutaneous leishmaniasis is a parasitic disease that causes ulcers on
exposed parts of the body and leads to lifelong scars. The World Health
Organization estimates that 0.7 to 1.3 million new cases of CL occur
annually, making it the most common form of leishmaniasis. In the
Old World, the vector for CL is the Phlebotomus spp. sand fly. The
primary reservoir host in Central Asia is the Great Gerbil (Rhombomys
opimus). Phlebotomus spp. sand flies were discovered in Bakanas,
Kazakhstan for the first time in June 2012. In light of this discovery, it
was determined that a field trial should be conducted to test the
efficacy of the insecticide, fipronil against Phlebotomus spp. sand
flies by feeding it to great gerbils in bait form. A field trial was
conducted in Bakanas, Kazakhstan in May and June 2013 to determine
the abundance of Phlebotomus spp. sand flies in the area. In addition,
the efficacy of a fipronil-based bait in controlling female sand fly
numbers was to be conducted. The study design involved monitoring
sand flies using CDC light traps for two weeks prior to treatment and
for four weeks post treatment on two treatment plots and two control
plots. Both treatment plots were treated with a one-time application of
fipronil based gerbil bait. Sand flies that were collected from CDC
light traps were sexed and identified down to the genus level for
Sergentomyia spp. and to the species level for Phlebotomus spp. Data
for total numbers of sand flies per trap night were recorded in an Excel
Spreadsheet. Two weeks of additional CDC light trapping were
performed in September 2013. The research resulted in some
interesting ecological findings. Nearly 100% of the sand flies
collected were Paraphlebotomus mongoliensis. Of particular interest
was the density of blood fed female sand flies. Of the 3,487 female P.
mongoliensis that were collected, 1,248 had taken blood meals (36%).
This number is abnormally high and the data could be very beneficial
to developing a sand fly control program in Kazakhstan. The efficacy
of the fipronil bait is yet to be determined.
Differently from visceral leishmaniasis (VL) caused by Leishmania
donovani, which is a disease transmitted only among people, VL
caused by L. infantum is a zoonosis transmitted from domestic and
wild canines. However, patients with VL coinfected with HIV or
immunocompetent are also known to infect the vectors Phlebotomus
perniciosus and Lutzomyia longipalpis. Besides, Lu. longipalpis is an
eclectic vector, including to humans, who are important blood
sources for this species in urban settings in Brazil. Therefore,
nevertheless neglected by control programs, the role of humans as
reservoirs of L. infantum seems plausible. This study aimed to
compare the competence of symptomatic and asymptomatically
infected humans, with and without HIV, as reservoirs of L. infantum.
The study groups were composed by (a) 21 patients with LV and HIV
(VLHIVs); (b) two asymptomatic patients with LV and HIV
(VLHIVa); (c) 21 patients with VL but without HIV (VLs); and (d) 19
asymptomatic individuals infected with L. infantum but without HIV
(VLa). All patients underwent xenodiagnosis with Lu. longipalpis.
Insects were dissected and examined by optical microscopy and by
conventional PCR for Leishmania spp. The infections ratios were
13/21 in VLHIVs; 1/2 in VLHIVa; 8/21 VLs: and 3/19 in VLa. The
results show that being symptomatic for VL increases infectivity of
humans, but also that asymptomatic individuals with or without HIV
infection are also able to transmit L. infantum to the vector Lu.
longipalpis. These results demonstrates that humans should be
considered as source of L. infantum when designing control
programs for urban VL in the New World.
N° 26 (1) 2015
ID 11-P Control
ID 12-P Ecology and epidemiology
ADDRESSING THE EMERGENCE AND SPREAD OF
LEISHMANIASIS IN THE BORDERS OF
ARGENTINA, BRAZIL AND PARAGUAY
MOLECULAR DETECTION OF LEISHMANIA IN
PHLEBOTOMINE SAND FLIES IN A CUTANEOUS
AND VISCERAL LEISHMANIASIS ENDEMIC AREA
IN NORTHEASTERN BRAZIL
Quintana María Gabriela1 ,2,.3 ,4, Santini María Soledad1 ,2 ,5, GonzálezBritez Nilsa6, Rojas de Arias Antonieta7, Celeste Vega8, Thomaz-Soccol
Vanete9, Bisetto Júnior Alceu10, Yadon E Zaida11, Salomón Oscar Daniel1,2,4
and the IDRC Project #107577 team.
Instituto Nacional de Medicina Tropical-MSN, Argentina; 2 REDILA,Argentina; 3
Instituto Superior de Entomología-UNT, Argentina; 4 CONICET, Argentina; 5Centro
Nacional de Diagnóstico e Investigaciones en Endemoepidemias-ANLIS-MSN,
Argentina; 6 Facultad de Ciencias de la Salud-Universidad Nacional del Este,
Paraguay; 7 OPS/OMS Paraguay, 8 Centro para el Desarrollo de la Investigación
Científica (CEDIC), Asunción, Paraguay. 9 Departamento de Engenharia de
Bioprocessos e Biotecnologia, Univerasidade Federal do Parana, Brazil; 10 Secretaria
de Saúde do Estado do Paraná, Brazil, 12 Communicable Diseases Department, Pan
American Health Organization, Washington, D.C., United States of America
65
Vanessa Cristina Fitipaldi Veloso Guimarães, Pietra Lemos Costa,
Fernando José da Silva, Fábio Lopes de Melo, Filipe Dantas-Torres,
Eduardo Henrique Gomes Rodrigues and Sinval Pinto Brandão-Filho
1
Keywords: Eco-Health, borders, multidisciplinary
In the border region of Argentina, Brazil and Paraguay, cutaneous
leishmaniasis generates rural and peri-urban outbreaks strongly
associated with environmental and anthropogenic change Facultad de
Ciencias de la Salud-Universidad Nacional del Este, Paraguay.s, while
visceral leishmaniasis is emergent and spreading in urban and periurban areas. Therefore, different forms of transmission of
leishmaniases occur in a diversity of settings, ranging from forest to
urban, and from work to peri-domestic associated risks. The region
also has a wide diversity of human populations, including different
cultural backgrounds, as native peoples, migrants (transient and
permanent settlements), and tourists. In order to address the
emergence and spread of leishmaniasis a project was designed to
identify the main biological, environmental and social drivers
associated with parasite transmission risks in different landscapes,
within the theoretical frame of Eco-Health. This project engages
researchers and decision makers from the three countries to respond to
the explicit demand of public health authorities to improve the
effectiveness of leishmaniasis control programs. The design involves
“mirror” activities in Argentina, Brazil and Paraguay, and
iterative/successive feedbacks instead of simultaneous analysis
between the results of the dialectic sciences (social) and cumulative
ones (bio-ecological). Up to now the project team consolidated the
historical information about the leishmaniases, and the sociohistorical context that allowed the transmission in the region, the
social scientists performed a base-line perception study, and the three
countries multidisciplinary specialists gathered in a workshop to
establish a common epistemological background and to refine the
field designs. The antecedents included the previous studies on
distribution in time and space of Phlebotomine, and its relation with
outbreaks and/or humans activities. In each country a specific set of
interventions will be developed. The participation of decision makers
from the local, subnational and national levels, the technical
collaboration of the Pan-American Health Organization and the
partnership of the International Development Research CentreCanada, will promote the uptake of results and implementation of the
proposed multi-sectorial prevention and control strategies, as well as
their eventual replication in other areas. Financial Support IDRCCanada Project #107577, Ministry of Health of Argentina, Brazil
Paraguay; PAHO.
Centro e Pesquisas Aggeu Magalhães- Fiocruz
vanessa@cpqam.fiocruz.br
Keywords: Phlebotomine sand flies, Leishmania; transmission
Several phlebotomine sand fly species have been regarded as putative
or proven vectors of parasites of the genus Leishmania in Brazil, but
data for the northeastern region remains incipient. In this study, a total
of 600 phlebotomine sand flies were grouped in pools of 10
specimens each and tested by a Leishmania genus-specific PCR and
by a PCR targeting Leishmania (Leishmania) infantum. Fourteen out
of 60 pools were positive by the genus-specific PCR, being five pools
of Lutzomyia migonei, seven of Lutzomyia complexa, one of
Lutzomyia sordellii and one of Lutzomyia naftalekatzi, which
corresponds to a minimal infection rate of 2.3% (14/600). Our results,
associated with their known anthropophily and their abundance,
suggest the participation of L. migonei and L. complexa as vectors of
Leishmania in northeastern Brazil. Remarkably, the detection of
Leishmania DNA in L. sordellii and L. naftalekatzi is reported for the
first time in this country, but future studies are necessary to better
understand the significance of these findings.
N° 26 (1) 2015
ID 13-P Ecology and epidemiology
ID14-P Ecology and Epidemiology
STUDY OF SAND FLY FAUNA IN RECENT AREA
TRANSMISSION OF LEISHMANIASIS IN THE STATE
OF MINAS GERAIS, BRAZIL
N E W R E C O R D A N D U P D AT E
GEOGRAPHICAL DISTRIBUTION OF
MONTICOLA (COSTA LIMA, 1932)
PSYCHODIDAE: PHLEBOTOMINAE)
AMERICA
Menezes, Júlia Alves1; Sousa, Alessandra Mara2; Morais, Mayron
Henrique Gomes2; Andrade-Filho, José Dilermando1; Lima, Fernanda
Pinheiro3; Fux, Blima4; Margonari, Carina1
1
2
66
ON THE
PINTOMYIA
(DIPTERA:
IN SOUTH
Andrey José de Andrade & Rodrigo Gurgel-Gonçalves
Centro de Pesquisas René Rachou, Brazil; Fundação Educacional de Divinópolis,
Brazil; 3Centro Universitário de Formiga, Brazil; 4Universidade Federal do Espirito
Santo, Brazil
Laboratório de Parasitologia Médica e Biologia de Vetores, Área de Patologia,
Faculdade de Medicina, Universidade de Brasília, Campus Universitário Darcy
Ribeiro, Asa Norte, 70910-900, Distrito Federal, Brazil.
jandrade@cpqrr.fiocruz.br
bioandrey@gmail.com
Keywords: Lutzomyia longipalpis, Lutzomyia cortelezzii, Lutzomyia
whitmani, Visceral Leishmaniasis
Keywords: Pintomyia monticola, potential distribution, Federal
District, ecological niche modeling
Introduction: Study the sand fly fauna in recent foci of leishmaniasis
is crucial to know the risk of disease transmission, since it allows to
clarify the distribution and vectors ecology. In the city of Formiga,
Minas Gerais, Brazil, the first cases of American visceral
leishmaniasis (AVL) occurred in 2011 and 2012, although still have
not been recorded american cutaneous leishmaniasis (ACL) cases.
Thus, the present study aimed to know the local sand fly fauna and
identify potential areas of transmission risk. Methodology: Monthly
systematic collections in the 24 peridomicile houses, using HP light
traps, ocurred for 4 consecutive days (5pm - 7am) between May 2012
to April 2013. The domiciles were selected in accordance with the
location of human and canine cases of leishmaniasis and the
peridomicile conditions. The insects captured were identified by
Young and Duncan key (1994). The Pearson correlation and chisquare tests were used to test the association of phlebotomines with
bioclimatic factors (humidity, temperature and rainfall). Results: Out
of 224 specimens were captured from 10 species, the most prevalent
were Lutzomyia longipalpis (n=79, 35.3%), Lutzomyia cortelezzii (n =
75, 33.5%) and Lutzomyia whitmani (n=41, 18.3%). The first two
presented widely distributed by the municipality (22 and 18 collection
points, respectively), while L. whitmani concentrated in just 4 points,
especially near the green areas. A significant correlation between
environmental conditions and abundance of sandflies were observed.
It was found a high correlation between sandflies and rainfall
(r2=0.52, p=0.008) and a moderate correlation between
phlebotominaes and temperature (r2= 0.36, p=0.039). Conclusions:
The occurrence of AVL cases and the finding of L. longipalpis indicate
that probably this species is associated with the transmission of the
leishmaniases in the city. This hypothesis agree with the fact that L.
longipalpis was found in almost all collection points, but more studies
are necessary. As for L. whitmani, even without records of ACL cases,
their finding indicates the necessity of local entomological
surveillance, due to the involvement of this species in the disease cycle
. Finally, the near quarters with preserved forest areas, where the
presence of L. whitmani were abundant, may exhibit a higher
vulnerability to the appearance of the first cases of ATL.
Pintomyia monticola was described based on five females captured in
the state of São Paulo and males were described using specimens from
a laboratory colony. The species have been recorded in Peru,
Argentina, Paraguay and Brazil. It is a suspected vector of
Leishmania enriettii, the etiological agent of cutaneous leishmaniasis
in guinea pigs, in southern Brazil. The present study updates the
distribution of P. monticola and presents a potential geographical
distribution model for this species. Distributional data were obtained
from various published studies and speciesLink data
(http://splink.cria.org.br). When only textual georeferences were
provided they were georeferenced based on internet resources
(http://www.fallingrain.com/world/). Records were georeferenced
with an uncertainty of ≤5 km to the nearest 0.01° (records presenting
greater uncertainty were removed). Data were separated into two sets:
one for model calibration (75% of points) and one for model
evaluation (25% of points). Eight climatic variables from the
WorldClim project were obtained to characterize climate variation
across the distribution range of P. monticola. All environmental
databases used in this study covered areas of South America,
resample to a spatial resolution of 2.5' (~5 km). Ecological niche
models were produced using Maxent version 3.2.1. We used median
output grids as a hypothesis of suitability, and imported the results
into ArcView 3.3 for analysis. We assessed model accuracy by
examining omission rate associated with test points. We also used
Maxent's jackknife test to identify variables that most influenced
model predictions. P. monticola was registered in 143 localities and
its updated geographical distribution map showed the bona fide
records in South America and the new record (seven females) in
Brasília, Federal District, Brazil. When niche models were displayed
in geographical space P. monticola showed a wide potential
distribution in Brazil passing through Bolivia and reaching Peru. All
test points were included in the predicted suitable area (0% omission).
Highest climate suitability for P. monticola was observed along the
Brazilian Atlantic coast. When contribution of climatic variables was
explored, annual mean temperature, temperature seasonality, and
minimum temperature in the coldest month were the variables that
most influenced models of P. monticola. The results presented here
extended the geographical distribution of P. monticola in South
America.
N° 26 (1) 2015
67
ID 15-P Ecology and Epidemiology
ID 16-P Ecology and Epidemiology
E N TO M O L O G I C A L M O N I TO R I N G O F T H E
PROGRAMME OF VISCERAL LEISHMANIASIS IN
MACEIÓ, ALAGOAS STATE, BRAZIL: INDICATORS
AND ENVIRONMENTAL RISK FACTORS
ASPECTS OF THE ECOLOGY OF LUTZOMYIA
LONGIPALPIS AND LUTZOMYIA MIGONEI (DIPTERA:
PSYCHODIDAE: PHLEBOTOMINAE) IN AREA OF
INTENSE TRANSMISSION OF VISCERAL
LEISHMANIASIS IN NORTHEAST OF BRAZIL:
FORTALEZA, STATE OF CEARÁ, BRAZIL
Carlos Fernando Rocha Dos Santos; Magliones Carneiro Lima;
Grimoaldo Braga Da Rocha Neto; Ana Lúcia Moreira Macena
Secretaria de Saúde de Maceió. Alagoas. Brazil
cfr_santos@hotmail.com
Rafaella A. Silva, Claudia M. L. Bevilaqua, Elizabeth F. Rangel, Fabricio
K. M. Santos, Lindemberg C. Sousa. Lucas Edel Donato, Grasielle C. D.
Pessoa
Ministério da Saúde do Brasil
Keywords: Maceió, Phlebotominae, Lutzomyia
The city of Maceió is ranked, according to the methodology of the
Ministry of Health as sporadic transmission to American Visceral
Leishmaniasis (AVL). The occurrence of cases follows the process of
human occupation, located in discontinuous areas and sparse, with
half of the territory is comprised of rural areas, integrating even this
scenario, a park and an area of environmental preservation.
Entomological surveillance actions were triggered initially aimed to
determine the distribution of the vector, Lutzomyia (Lutzomyia)
longipalpis through entomological survey and monitoring, in order to
know the seasonality. Monthly collections were made with the help of
CDC light traps in the period from July 2004 to December 2012, and
selected areas for implementation of entomological monitoring from
identifying the points of greatest abundance of sand flies in the
entomological survey, correlated to the presence of human and canine
cases of the disease. The traps were installed from 18h and 07h drawn
around the next day for three consecutive nights, totaling 20 traps
indoors and peridodomestic sites. We collected 16,765 sand flies and
recorded nine species, especially L. (Nyssomyia) whitmani (58%), L.
(Lutzomyia) longipalpis (30%) and L. migonei (10%), all other
species amounted to 2%. The locations of the Povoado Canto do Farias
and Sitio São Gonçalo and Riacho Doce were those which had the
highest abundance vector. Entomological indicators showed variation
according to the presence of domestic animal shelters around the
houses, and also regarding the conditions of residence and can be seen
a decrease in vector abundance inside the house after improvements
made. The results showed that there was a relationship between sand
fly density and housing conditions and environmental factors,
especially the presence of domestic animals (chickens and pigs).
Environmental characteristics and lack of adequate infrastructure in
communities, and how precarious proximity with the forest, presence
of pets around the houses, garbage accumulation, favored the
maintenance of sand fly vector breeding sites, increasing the risk of
disease transmission. The entomological data found allies to
environmental risk factors identified in the area suggest the need to
consider in an integrated manner, entomological indicators and
environmental management as a condition for effective control
actions.
rafaas28@gmail.com
Keywords: Lutzomyia longipalpis, Lutzomyia migonei, Visceral
leishmaniasis, Fortaleza
The main vector for visceral leishmaniasis (VL) in Brazil is
Lutzomyia longipalpis. However, the absence of L. longipalpis in a
region of autochthonous VL demonstrates the participation of other
species in the transmission of this disease. Studies conducted in La
Banda, Argentina, and São Vicente Férrer, Brazil, have correlated the
absence of L. longipalpis and the presence of L. migonei with
autochthonous cases of VL. In São Vicente Férrer, there was evidence
for natural infection of L. migonei with Leishmania infantum chagasi,
which strongly supports the participation of this vector in disease
transmission in the area. Thus, the objective of this work was to assess
the ecology of the sand flies L. longipalpis and L. migonei in
Fortaleza, an endemic area for VL. Insect capture was conducted at 22
sampling points that were distributed across four regions of Fortaleza,
and two CDC light traps were used at each collection point. In total,
32,403 sand flies were captured; of these, 18,166 (56%) were
identified as L. longipalpis and 14,237 (44%) as L. migonei. There
were significant differences found between the vectors in each
sampling site (indoors and outdoors) (p <0.0001). These findings
confirm that L. migonei and L. longipalpis are distributed throughout
Fortaleza, where they have adapted to an indoor environment.
Because these species have been shown to be competent vectors for
VL, our results suggest that L. migonei shares the role of vector with
L. longipalpis for the transmission of VL in Fortaleza.
N° 26 (1) 2015
ID 17-P Ecology & Epidemiology
PHLEBOTOMINAE CAPTURED IN AREA OF
TRANSMISSION OF AMERICAN CUTANEOUS
LEISHMANIASIS (ACL) IN XAPURI MUNICIPALITY,
ACRE STATE, WESTERN BRAZILIAN AMAZONIAN.
PRELIMINARY RESULTS
THE GEOGRAPHIC EXPANSION OF AMERICAN
VISCERAL LEISHMANIASIS ASSOCIATED WITH
DEFORESTATION IN TOCANTINS STATE, BRAZIL
Andreia Fernandes Brilhante, Eunice Aparecida Bianchi
Galati, Cristiane de Oliveira Cardoso, Nathanna Progênio
dos Santos, Jailson Ferreira de Souza and Joaquim Vidal
Faculdade de Saúde Pública da Universidade de São Paulo / Faculdade Meta Grupo
Athenas Educacional
brilhanteaf@usp.br
Keywords: Sand flies, vectors, american cutaneous leishmaniasis,
Amazonia
The study aimed identify the phlebotomine fauna in the Xapury
municipality. The sand flies were captured monthly from August 2013
to February 2014 with automatic light traps (CDC type) from 6:00pm
to 06:00am in rural and urban areas. Twelve traps were installed in
forests, peridomiciles (in domestic animal shelters) and
intradomiciles. The insects were macerated and identified in
accordance with Galati (2003). A total of 3,064 specimens (1,065
males and 1,999 females) have been captured. They belong to 47
species: Bichromomiya flaviscutellata (0.07%), Brumptomyia sp
(0.10%), Br. avellari (0.03%), Br. penthacanta (0.19%), Evandromyia
bacula (0.03%), Ev. begonae (0.13%), Ev. infraspinosa (0.10%), Ev.
saulensis (1.86%), Ev. termitophila (0.10%), Ev. walkeri (0.03%),
Lutzomyia sherlocki (0.22%), Migonemyia migonei (0.03%),
Nyssomyia antunesi (0.65%), Ny. richardward (0.07%), Ny. shawi
(23.3%), Ny. whitmani (0.62%), Ny. yuillii yuillii (0.59%),
Psathyromyia aragaoi (0.30%), Pa. campbelli (0.03%), Pa. cuzquena
(0.03%), Pa. dendrophila (0.20%), Pa. punctigeniculata (0.03%), Pa.
(Forattiniela) sp (0.10%), Pa. serie Shannoni (0.03%), Pintomyia
nevesi (0.65%), Pi. serrana (0.03%), Pressatia sp (0.40%), Pr. choti
(0.16%), Pr. tricantha (0.03%), Psychodopygus serie Guyanensis
(0.62%), Ps. amazonensis (0.13%), Ps. carrerai carrerai (9.6%), Ps.
claustrei (0.13%), Ps. davisi (9.14%), Ps. llanosmartinsi (8.25%), Ps.
hirsutus hirsutus (2.61%), Ps. lainsoni (1.80%), Ps. paraensis
(0.13%), Ps. yucumensis (0.35%), Sciopemyia sordellii (0.10%),
Thrichophoromyia auraensis (10.20%), Th. octavioi (19.50%), Th. sp.
(5.6%), Th. sp. n (0.13%), Th. ubiquitalis (1.40%), Trichopygomyia
sp. (0.07%) and Viannamyia furcata (0.13%). About 95% of the all
specimens collected belong to the three genera: Nyssomyia (25.2%),
Psychodopygus (32.8%) and Trichophoromyia (36.9%). Four
specimens of Trichophoromyia of a species not yet described were
found. Of the two predominant genera Nyssomyia and
Psychododpygus there are several anthropophilic species and some of
them suspect of transmit ACL agents and Th. auraensis is the proved
vector of Leishmania lainsoni. The occurrence almost exclusively of
the sand flies in rural area with vector species suggest that the
transmission of ACL agents has occurred in this environment. The
residents and visitors should be reminded, especially because one of
the investigated points has attracted tourists from the region and other
areas including from abroad.
68
Margarete Martins dos Santos Afonso1, Sérgio Augusto de Miranda
Chaves 2, Mônica de Avelar Figueiredo Mafra Magalhães 3, Renata
Garrido 3, Carina Graser 41, Bruno M. de Carvalho1, Simone M. Costa 1,
Elizabeth Ferreira Rangel
1
Laboratório de Transmissores de Leishmanioses, Laboratório de Referência em
Vigilância Entomológica: Taxonomia e Ecologia de Vetores das Leishmanioses,
Instituto Oswaldo Cruz Fundação Oswaldo Cruz; 2Laboratório de Ecologia,
Departamento de Endemias, Escola Nacional de Saúde Pública Sérgio Arouca Fundação Oswaldo Cruz; 3Laboratório de Geoprocessamento, Instituto de
Comunicação e Informação Científica e Tecnológica em Saúde; 4Secretaria de Saúde
do Estado do Tocantins.
mafonso@ioc.fiocruz.br
Keywords: Lutzomyia (L.) longipalpis, Epidemiology, Visceral
Leishmaniasis, Spreading and Urbanization
American Visceral Leishmaniasis (AVL) is an important Public
Health issue in Brazil, with evident geographic expansion to urban
areas of medium and large cities. Over the last decade, increasing
levels of environmental changes have been recorded in Tocantins
State, associated with agriculture and hydroelectric plant
constructions. This scenario is observed in some regions of the state
where AVL is present. Currently the disease is recorded in 120 of 139
municipalities. This study aimed to evaluate the AVL expansion in
Tocantins State, by associating the deforestation increase with AVL
stratification, as well as the presence of AVL with deforestation from
2004 to 2011. Annual deforestation rates in the Amazon region were
obtained from the PRODES project (National Institute for Space
Research, INPE). Disease stratification data was provided by the
Health Department of Tocantins State. The software ArcGIS 10 was
used for spatial analysis and map production. Chi square tests
(confidence interval 95-99%) were applied with software SPSS 19 to
evaluate the association between human cases of AVL and presence of
deforestation. From 2004 to 2011, the state's northern region showed
a constant increasing in deforestation and high numbers of AVL
human cases, especially in the municipalities of Araguaína,
Araguatins, Bandeirantes do Tocantins, Arapoema, Santa Fé do
Araguaia, Ananás, Pau d'Arco, Itaporã do Tocantins, Nova Olinda,
Colméia, Colinas do Tocantins and Riachinho. The results showed an
association between deforestation and AVL transmission, with
significant correlation values in 2005 (p=0.119), 2007 (p=0.114),
2008 (p=0.218), 2009 (p=0.039), 2010 (p=0.013), and 2011
(p=0.004). This confirms the hypothesis that the deforestation favors
disease transmission. Most municipalities with AVL human cases had
also deforestation records. The increase of the statistical significance
over the last three years (2009-2011) is noteworthy. In Tocantins
State, the analysis showed an association between deforestation and
AVL transmission, corroborating studies which associate
deforestation to epidemic outbreaks. In Brazil, AVL human cases are
increasingly being recorded in environmentally changed areas. The
results suggests the need for integrated planning and development of
public policies to minimize the impacts of human-made
environmental changes in AVL transmission. Financial support:
IOC/FIOCRUZ, CAPES, CNPq (PAPES VI, 407707/2012-3),
FAPERJ (E-26/110.525/2012).
Edición especial (marzo 2015)
Financial support: IOC/FIOCRUZ, CAPES, CNPQ (PAPES VI,
407707/2012-3), FAPERJ (E-26/110.525/2012).
LUTZOMYIA (LUTZOMYIA) LONGIPALPIS FEEDING
HABITS AND NATURAL INFECTION BY LEISHMANIA
(LEISHMANIA) INFANTUM CHAGASI IN SOME
BRAZILIAN ENDEMIC AREAS OF AMERICAN
VISCERAL LEISHMANIASIS
Margarete Martins dos Santos Afonso1, Sérgio Augusto de Miranda
Chaves2, Daniela de Pita Pereira3, Taiana Amâncio da Costa Rego3,
Antônio Luis Ferreira de Santana1, Júlia dos Santos Silva1, Bruno
Moreira de Carvalho1, Simone Miranda da Costa1, Cristina Giordano4,
Elizabeth Ferreira Rangel1
1
Laboratório de Transmissores de Leishmanioses, Laboratório de Referência em
Vigilância Entomológica: Taxonomia e Ecologia de Vetores das Leishmanioses,
Instituto Oswaldo Cruz - Fundação Oswaldo Cruz; 2Laboratório de Ecologia,
Departamento de Endemias, Escola Nacional de Saúde Pública Sérgio Arouca Fundação Oswaldo Cruz; 3Laboratório de Biologia Molecular e Doenças Endêmicas Fundação Oswaldo Cruz, 4 Secretaria de Saúde do Estado do Rio de Janeiro.
mafonso@ioc.fiocruz.br
69
SAND FLIES (DIPTERA, PSYCHODIDAE,
PHLEBOTOMINAE) COLLECTED IN THE WAJÃPI
INDIGENOUS RESERVE, EASTERN AMAZON,
BRAZIL
Eduardo Stramandinoli Moreno1; Maíra Posteraro Freire2,4, Luis Otávio
Cordeiro Nascimento3, Volmir Zanini3, Veracilda Ribeiro Alves4, Paloma
Helena Fernandes Shimabukuro4
1
DSEI Amapá e Norte do Pará – Secretaria Especial de Saúde Indígena /Ministério da
Saúde. 2ILMD–PPG Saúde, Sociedade e Endemias da Amazônia (UFAM/FIOCRUZAmazônia/UFPA) 3Laboratório Central de Saúde Pública do Amapá–LACEN/AP.
4
Centro de Referência Nacional e Internacional para Flebotomíneos-Centro de
Pesquisas René Rachou/FIOCRUZ.
palomahfs@gmail.com
Keywords: Lutzomyia (L.) longipalpis, Leishmania (L.) infantum
chagasi, Visceral Leishmaniasis, Ecology
Brazil faces the geographical expansion and urbanization of American
Visceral Leishmaniasis (AVL). The presence of the vector, Lutzomyia
(Lutzomyia) longipalpis, in urban areas is one of the greatest
challenges for Brazilian Visceral Leishmaniasis Control Program. The
knowledge on the biology of L. (L.) longipalpis can help to develop
more appropriate and effective control strategies as well as preventive
actions. The study aimed to combine diagnostic molecular tools to
detect, simultaneously, natural infection by Leishmania (Leishmania)
infantum chagasi and the feeding source of L. (L.) longipalpis. Sand
fly specimens were captured in the following Brazilian municipalities:
Araguaína (Tocantins State), Fortaleza (Ceará State), Sobral (Ceará
State) (the three with high incidences of AVL) and Rio de Janeiro (Rio
de Janeiro State) (first urban focus of the city). Molecular diagnostics
were performed by Polymerase Chain Reaction and Dot-Blot
Hybridization with primers for Homo sapiens, Canis familiaris,
Didelphis albiventris, Gallus gallus and L. (L.) infantum chagasi. The
evaluation of the feeding source had a general positivity index of
29.3%. Lutzomyia (L.) longipalpis showed high positivity for humans
(61.5%), followed by dogs (28.2%) and birds (10.3%). In Araguaína
municipality, most of the samples were positive for humans (61.5%)
followed by dogs (38.5%). In Fortaleza and Sobral, specimens had
higher positivity for humans (54.5% and 66.7%), followed by dogs
(27.3% and 20.0%) and birds (18.2% and 13.3%). The detection of
feeding source from specimens collected in Rio de Janeiro was not
possible, maybe because the specific primer was not used. The
evaluation of natural infection had a general positivity index of 4.61%,
with 4.9% from Araguaína and 9.75% from Sobral specimens. The
simultaneous detection of feeding source and natural infection had the
following results: in Araguaína, 2 specimens were naturally infected,
one with dog blood and one with human blood; in Sobral, 3 specimens
were naturally infected and had ingested human blood. Results
corroborate previous demonstrations of the feeding plasticity and
opportunistic behavior of L. (L.) longipalpis: it feeds on dogs,
domestic reservoirs of L. (L.) infantum chagasi; on birds, animals
whose attract sand flies and facilitate their domiciliation; and on
humans, which reinforces its anthropophilic habit.
Keywords: Amazon, Cutaneous leishmanises, Phlebotominae,
Vectors
To contribute to the understanding of the epidemiology of American
cutaneous leishmaniasis (ACL) among indigenous people in Eastern
Brazilian Amazon entomological collections were carried out at the
Wajãpi Indigenous People Reserve in the state of Amapá, Brazil. The
Wajãpi people consist of a group of approximately 1,000 indigenous
people living in a remote area of the Brazilian Amazon forest, close to
the border with French Guyana. The Wajãpi are subsistence
agriculturalists who also hunt and collect forest products such as fruits
and seeds. Life in the forest puts them in contact with the transmission
cycles of ACL. Sand flies were collected during the dry season
(September 2013) and rainy season (January 2014) with a total of 46
light traps distributed in 16 communities where cases of ACL have
been reported. The traps were set from 16:00-07:00hs, in intra and
peridomicile areas, and one area used frequently by the Wajãpi, such
as rivers, gardens or hunting trails. A total of 6,150 sand flies have
been collected. A total of 14 species have been identified so far, the
most abundant species are Trichopygomyia trichopyga (56.3%),
Psathyromyia dreisbachi (26.4%), Trichophoromyia ubiquitalis
(3.8%), Bichromomyia flaviscutellata (3.8%), Evandromyia
infraspinosa (3.4%) and Th. brachipyga (1.9%). The remaining
species represented 4.4% of the total amount collected and they were:
Nyssomyia umbratilis, Pa. aragaoi, Th. ininii, Sciopemyia sordellii,
Sc. fluviatilis, Pa. inflata, Psychodopygus squamiventris maripaensis
and Ty. depaquiti, the latter is recorded for the first time in Brazil.
Three medically important sand fly species were collected Ny.
umbratilis, Ps. squamiventris maripaensis and Bi. flaviscutellata.
Sand flies were found in all areas sampled in both seasons suggesting
ACL can be transmitted in the intra and peridomicile areas. In
addition to sand fly collections, we have assessed social and
environmental variables such as presence of wild and domestic
animals, waste management, distance to rivers, gardens and forest
areas, etc. which will help in the identification of ecological patterns
for sand fly occurrence as well as the in the evaluation of hot spots for
ACL transmission among the Wajãpi people.
N° 26 (1) 2015
70
LUTZOMYIA LONGIPALPIS (LEISHMANIA INFANTUM
VECTOR) AND NYSSOMYIA WHITMANI
(LEISHMANIA BRAZILIENSIS VECTOR) IN THE CITY
OF PUERTO IGUAZÚ, NORTHEASTERN
ARGENTINA: SEASONAL ABUNDANCE AND
RELATIONSHIP WITH DOMESTIC ANIMALS
GEOGRAPHICAL DISTRIBUTION OF NYSSOMYIA
BARRETTO, 1962 IN BRAZIL
Caroline Amaral Machado, Veracilda Ribeiro Alves, Paloma Helena
Fernandes Shimabukuro
Centro De Pesquisas Rene Rachou/Fiocruz Minas
Fernández María Soledad ; Santini María Soledad ; Cavia Regino ; and
Salomón Oscar Daniel1,2
palomahfs@gmail.com
REDILA 1Instituto Nacional de Medicina Tropical, 2Consejo Nacional de
Investigaciones Científicas y Técnicas, 3Centro Nacional de Diagnóstico e
Investigaciones en Endemoepidemias 4Dpto. de Ecología, Genética y Evolución,
FCEN, UBA and Instituto de Ecología, Genética y Evolución de Buenos Aires UBACONICET. odanielsalomon@gmail.com
Keywords: Cutaneous leishmaniasis, vectors, Nyssomyia umbratilis,
Nyssomyia whitmani
1,2
3
4
Keywords: Sandflies, urban, peridomiciles, ecology
In this study the city of Puerto Iguazú was sampled from spring 2011 to
winter 2012 to evaluate seasonal abundance changes of sandflies. The
city was divided in 51 areas of 400x400m where minilight traps were
located at the beginning of every season during two consecutive nights
in peridomiciles of households. Lutzomyia longipalpis and Nyssomyia
whitmani represented the 97.5% of the captures (N=5409 sandflies).
Two abundance indexes were analyzed for each species: the
occurrence and the number of individuals captured by night in each
household. The seasonality of occurrence and abundance by species
was evaluated using Generalized Linear Mixed Models. We also
evaluate the effect of the number of dogs and chickens on the
occurrence of sandflies at each household. For both analyses,
household were considered as a random effect. Both species showed
differences by season in the number of households-occurrence and
number of individuals captured, but with different patterns. The
percentage of households with Lu. longipalpis was higher in fall (66.7
%) compared to spring (31.4%) and summer (27.5%), and lower in the
winter (10.2%). The percentage of households with Ny. whitmani was
minimum in fall (9.8%) and was maximum in summer (39.2%) and
spring (31.4%), while winter has an intermediate value (22.5%). The
occurrence of both species was positively associated with the number
of dogs and chickens in the households. For Lu. longipalpis also a
synergic effect was detected in the number of dogs and chickens, while
for Ny. whitmani the effect of the number of chickens varied among
seasons. The percentage of households infested with both species was
maximum at the spring (21.5%) and summer (23.5%). The number of
individuals captured of both species changed according to the seasons
similarly to the occurrence. Lu. longipalpis abundance was higher in
fall (27.8 Phlebotomine per trap-night [PTN]) compared to spring and
summer (5.6 and 5.3 PTN) and the winter was the season with the
lower value (0.6 PTN). Ny. whitmani numbers were minimum in fall
and in winter (1.0 and 0.4 PTN) and were higher in summer and spring
(12.0 and 6.8 PTN). These results shows that in the study area these
sandflies have different patterns of abundance according to the
seasonality and also the numbers of domestic animals in the
households as chickens or dogs, and so the seasonal and spaceenvironmental risk is different for cutaneous leishmaniasis and
visceral leishmaniasis in the same city.
The genus Nyssomyia Barretto, 1962 comprises 20 species of which
14 are recorded in Brazil, including vectors of Leishmania in
American cutaneous leishmaniasis (ACL) transmission areas of
Central and South America. The aim of this study is to map the
geographical distribution of the species from the genus Nyssomyia in
Brazil and discuss the distribution of medically important species. A
dataset is being built based on literature search on PubMed, and
records obtained from examination of specimens from two
entomological collections: Coleção de Flebotomíneos of Centro de
Pesquisas René Rachou and Museu de Zoologia da Universidade de
São Paulo. So far, our results show the distribution of Nyssomyia
anduzei in the north region of Brazil with most records from
Amazonas state (AM), this species has been frequently associated
with wild and semi-domestic areas in the north region; N. umbratilis
has been mostly recorded from Pará state (PA), and more recently
records for this species are from AM, mainly from Manaus city and its
surroundings, this species has been found associated with humans and
domestic animals accidentally when in contact with transmission
cycles in the forest; N. delsionatali, N. richardwardi, N. shawi, N.
sylvicola, N. urbinatti, N. yuilli pajoti and N. yuilli yuilli present more
restricted geographical distribution in Brazil, mainly for the north and
central regions of Brazil; N. singularis has been only recorded in São
Paulo state (SP) from the type-locality suggesting it is not a valid
species; N. neivai is recorded for the southeast region. Most records
are for N. whitmani, in all regions of Brazil, especially in the southeast, including Minas Gerais state, where it has been found in as
diverse environments as urban, rural and forest areas, In SP it has
been recorded throughout the state. Nyssomyia antunesi has been
found in both wild and semi-domestic areas in the north and northeast regions, mainly in AM and PA states. Nyssomyia intermedia is the
most adapted species to anthropic environments, presents more
records for Rio de Janeiro and Espírito Santo states, but is found in all
regions of Brazil with the exception of the north region. Our compiled
data contributes to the knowledge of the geographical range of
Nyssomyia and shows where most work on ACL has been carried out
by researchers and public health workers.
N° 26 (1) 2015
VERTICAL AND HORIZONTAL DISTRIBUTION OF
SAND FLIES IN AN AREA OF TEGUMENTARY
LEISHMANIASIS IN CHACO, ARGENTINA
NATURAL BREEDING SITES OF PHLEBOTOMINAE
(DIPTERA: PSYCHODIDAE) IN A FLOOD ZONE OF
CHACO, ARGENTINA.
Rosa JR1, Parras MA1,2, Szelag EA1,2 Salomón OD2
71
Parras Matías Ariel1, 2, 3, Rosa Juan Ramón2, 3, Szelag Enrique Alejandro1, 2,
, Salomón Oscar Daniel1, 3
3
1
Laboratorio Entomología. Instituto de Medicina Regional. Universidad Nacional del
Nordeste. Chaco. Nodo REDILA (Red de Investigación de la Leishmaniasis en
Argentina). 2Instituto Nacional de Medicina Tropical (INMeT). Iguazú, Misiones.
Argentina.
1
Instituto Nacional de Medicina Tropical (INMeT), 2Instituto de Medicina Regional
(IMR - UNNE), 3Red de Investigación de la Leishmaniasis en Argentina (REDILA)
parrasmatias@gmail.com
juan_rosa05@yahoo.com.ar
Keywords: Stratification, Migonemyia migonei, Evandromyia
complex , Bionomy
During the active search for human cases of cutaneous leishmaniasis
in a rural area (26º20'22"S, 60º15'44") of Tres Isletas a house was
selected as worst epidemiological scenario. Five CDC-type light
traps/month/night/12hs(19:00 to 7:00 pm) were used, in domicile,
peridomicile (goats pens and piggery) and forest 110m from domicile
where deforestation and anthropic activity were evident. Also in the
forest two traps were installed in a tree, near the ground on 1.5 m and
canopy level 10m. From December to December 2010-2012 a total
1179 Phlebotominae were captured: Migonemyia migonei (82%),
Evandromyia cortelezzii (2.4%) only males, Ev. sallesi (5%) only
males, Cortelezzii complex females (9%): (including Ev. cortelezzii
and Ev. sallesi of similar morphology), Nyssomyia neivai (0.3%),
Psathyromyia shannoni (0.1%) and Brumptomyia brumpti (1.2%).
The most abundant was Mg. migonei on December 2011 (n=309) with
average temperature and relative humidity of 31ºC and 32%,
respectively. Only in summer 2010 (November/December) all species
were recorded in peridomicile and domicile with the greatest
abundance in the first (n=467) and the lowest in the second (n=81)
predominating Mg. migonei in both ecotopes (n=405; n=71,
respectively). In vertical strata, all species were recorded in ground
level. In 2010, the highest abundance was canopy (n=289) in autumn
(March/April), and the lowest in ground level (n=245) in summer
(November/December). Migonemyia migonei was the predominant
specie followed by Ev. cortelezzii, Ev. sallesi and Cortelezzii complex
females. The least abundant were Pa. shannoni only in ground level
and Br. brumpti in ground and canopy levels. During all seasons 2010
there was Phlebotominae in both strata, the highest abundance was in
summer (n=422) with averages of temperature and relative humidity
of 28°C and 45% and lower in winter (n=20), 12°C and 55%. In 2011,
the highest was in spring (n=166) 24°C and 66% and the lowest in
summer (n=74) 27°C and 58%. Species suspected as vectors of
leishmaniasis in this region were recorded in this study in both strata in
all seasons. This behavior was probably due to the meteorological and
microclimate conditions, soil type and vegetation, limited by
physiological factors such as changes in the food habits favored by
anthropic activities. This highlights the need for further large-scale
studies to contribute to the knowledge of bionomics and
environmental change in an epidemiological context.
Keywords: Nissomyia neivai, Lutzomyia longipalpis, soil samples,
emergence traps
Phlebotomine preimaginal sampling was performed from August
2011 to April 2014 in Puerto Antequeras (27° 26'34"S, 58° 50' 57" W)
where adults of Lutzomyia longipalpis and Nissomyia neivai where
already reported as the main vectors of visceral and cutaneous
leishmaniasis, respectively. Two homes were selected, with the
criteria of 'worst scenario'. From August 2011 to March 2013
substrate samples 10cm diameter by 3cm deep were taken monthly;
and from April 2013 until April 2014 emergence traps modified from
Casanova (2001) were installed. In each home three ecotopes were
selected: domestic (external walls of the houses), peri-domestic (hen
houses and kennels) and extra-domestic (base of trees and fallen logs
and exposed roots). In the latter ecotope three points (trees) were
selected at 100m distant from each other, along transect from the
domicile. The collected soil samples were kept at 24±5ºC and
80±10% relative humidity and observed daily for 60 days in search of
the emergence of imagos for identification of species level. After this
period the remaining of the samples were processed by flotation with
supersaturated solution of NaCl and observed under
stereomicroscope looking for larvae, pupae, dead adults and/or pupal
exuviuae. The emergence traps were controlled every 15 days to
renew the sticky paper. Each trap remained in the soil for 60 days and
then was moved to another point within the same ecotope and
mictrohabitat considered as potential breeding site of sand-fly. A total
of 950 substrate samples were taken. A male of Ny. neivai was
obtained during flotation of a sample taken from the forest 100 meters
from the house, at the base of a fallen tree in January (summer). As a
result of the emergence traps 5 specimens of Ny. neivai were obtained
(2 males and 3 females) distributed in four out of the forty traps. One
was located in peridomestic ecotope (hen house) and three in the
forest (base of a tree). Of the specimens obtained, 4 were in months
before an extraordinary flood occurred between June and July 2013
and 1 was recovered nine months after that flood (April 2014). A
production of 3.60 phlebotominae/100m2/day was estimated. These
preliminary results showed the fitness of the floodplain area of the
Parana River in the province of Chaco for natural breeding sites of Ny
neivai. These studies are continued broadening the environmental
range to support the hypothesis and looking for the breeding sites of
Lu. longipalpis.
N° 26 (1) 2015
72
ANALYSIS OF SAND FLY (DIPTERA: PSYCHODIDAE)
BLOODMEALS USING SEQUENCING TECHNIQUE:
TEST OF SAND FLY EXPERIMENTALLY FED
SANDFLY SURVEY IN ECOTOURISM REGION IN
THE DISTRICT OF BOM JARDIM FROM THE CITY
OF NOBRES, MATO GROSSO - BRAZIL
Carvalho GML; Quaresma PF; Gontijo CMF; Rego FD; Rugani JMN;
Dias TA; Andrade Filho JD
Anagela Maria Acel, Ana Lucia Maria Ribeiro, Rosina Djunko
Miyazaki, Jorge Senatore Vargas Rodrigues
Centro de Pesquisas Rene Rachou - Fiocruz Minas. gumayr@yahoo.com.br
Universidade Federal de Mato Grosso
anagelamaria@bol.com.br
Keywords: Phlebotominae, Bloodmeal Molecular Biology
The eco-epidemiological factors involved in the genesis of different
forms of leishmaniasis in the New World are very complex. Several
species of Leishmania are capable of causing disease in humans which
in turn, interact with various sand fly species and mammalian hosts.
The result of these interactions is a profuse epidemiological variety
that difficult the understanding and control of the disease. Studies
considering the occurrence of different species of Leishmania in sand
flies have been essential for a better understanding of the ecoepidemiology of leishmaniasis. However, there are few studies
investigating the correlation between infected sand fly species and
their possible infection sources. Thus, the aim of this work is to
standardize the bloodmeal identification in experimentally engorged
females by sequencing technique, for future application in naturally
engorged females. For this study, we selected as target a fragment of
the Cytochrome B gene, present in birds and mammals and recently
used in other studies to identify the meal in hematophagous insects.
The tests were carried out using females of the first generation of a
sand fly colony and were exposed to the first meal in different
vertebrates. After the experimental fed, the females were sacrificed
and subjected to DNA extraction and polymerase chain reaction
technique (PCR) of the CytB target. Our results demonstrated a good
amplification of the fragment of 359bp in experimentally fed
specimens. The amplified products were purified and subjected to
sequencing and it was possible to identify each particular level
vertebrate host by aligning and comparing the sequences deposited in
GenBank. These first results were crucial for the next steps that are
being performed in attempt to optimize the technique and reproduce
the methodology in naturally engorged females. The identification of
the blood meal in naturally engorged females provides information
about food sources and may improve the knowledge about the
behavior aspects of sand flies like feeding preference or feeding
patterns in natural conditions. These informations could also
contribute to indicate the potential reservoir hosts, for Leishmania
species in different ecotopes. Financial Support: FAPEMIG/CAPES,
CPqRR/FIOCRUZ, CNPq.
Keywords: Phlebotomine, Leishmaniasis, Nobres-Bom Jardim/MT,
Ecotourism
The Leishmaniasis are infectious diseases, non-contagious, chronic
evolution caused by the protozoa of the genus Leishmania and
transmitted by the bite of insects called sandflies which belong to the
family Psychodidae, Genus Lutzomyia. The American Cutaneous
Leishmaniasis (ACL) and Visceral Leishmaniasis (VL) primarily
cause zoonotic infections, affecting humans and their pets. In Mato
Grosso there is a great diversity of sandfly species with reporting
cases in 100 % of the municipalities of ACL. The City of Nobres in the
State of Mato Grosso / Brazil, situated 142 km from the capital of
Mato Grosso, has natural wonders that attract more and more the
interest of tourists, highlighting the district of Bom Jardim by
numberless places for ecotourism. This study aimed to survey the
insect fauna of sandflies in the district of Bom Jardim. CDC light traps
were used, 8 (eight) indoor, 18 (eighteen) in peridomestic and 4 (four)
outdoor, all installed in homes and places with tourist attractions in
the months of December 2012 (starting rainy season); June/2013
(drought) and March/2014 (rainy season). A total of 465 sandflies
were captured, divided into two genera and 14 species as it follows:
Brumptomyia = 42 (9,0%); Lutzomyia antunesi = 171 (36,8%); L.
aragaoi = 3 (0,6%); L. carmelinoi = 42 (9,0%); L. davisi = 30 (6,5%);
L. evansi= 1 (0,2%); L. flaviscutellata = 5 (1,1%); L. hermanlenti = 22
(4,7%); L. lenti = 15 (3,2%); L. longipalpis = 3 (0,6%); L. walkeri =
116 (24,9%) e L. whitmani = 9 (1,9%). The proportion between male
and female was 52.7 % and 47.3 % respectively. Regarding the
collection environment, the percentage was: indoor=7.7%;
peridomestic=87.5% and 4.7% outdoor. Among the three collection
periods, the period that obtained a greater diversity of species and
greater quantity with a total of 92.47% of occurrence, was December.
Species that have implications in the transmission of leishmaniasis
are: L. flaviscultellata, L. whitmani and L. antunesi to ACL and L.
longipalpis to VL. Because this place is a great potential for
ecotourism, it should be monitored and a program of preventive
measures for leishmaniasis should be developed for both local people
and tourists.
N° 26 (1) 2015
LEISHMANIASIS SANDFLY DIVERSITY IN A RURAL
AREA OF THE CUIABÁ, MATO GROSSO, BRAZIL
INCIDENCE OF SANDFLIES AT THE ZOOLOGICAL
PARK OF FEDERAL UNIVERSITY OF MATO
GROSSO, CUIABÁ/MT-BRAZIL
Ana Lucia Maria Ribeiro1,2, Jorge Senatore Vargas Rodrigues1,2, Tiago
Parreiras de Almeida2, Emerson Marques de Souza3, Flávia Karolina
Pereira Barreto Bettiol3, Anagela Maria Acel1,2, Fabio Alexandre Leal
dos Santos1,2, Sirlei Franck Thies1,2, Rosina Dunjko Miyazaki3
1
- Julio Muller University Hospital, Faculty of Medicine, UFMT, Cuiabá, Mato Grosso,
Brazil 2 -Faculty of Medicine, UFMT, Cuiabá, Mato Grosso, Brazil 3 - Institute of
Biosciences, UFMT, Cuiabá, Mato Grosso Brazil
73
Rosina Dunjko Miyazaki, Ana Lucia Maria Ribeiro, Jorge Senatore
Vargas Rodrigues, Tiago Parreiras de Almeida, Emerson Marques de
Souza, Flávia Karolina Pereira Barreto Bettiol, Anagela Maria Acel
Universidade Federal de Mato Grosso
Keywords: Vector, Leishmaniasis, Zoological Park, Cuiabá/MT
Keywords: Phlebotomine, Leishmaniasis, Cuiabá/MT
Leishmaniasis is an infectious disease caused by protozoa of the genus
Leishmania, transmitted to mammalian hosts by insect vectors
belonging to Lutzomyia genus. Most of the infections are observed
among wild and domestic animals. Accidental zoonotic infections in
man occurs mainly in tropical areas, including in Brazil. According to
World Health Organization reports, endemic transmission of
leishmaniasis occurs in 98 countries, however there is no official data
for several others. The aim of this study was to investigate the variety
of sandfly species in a rural area of Cuiabá, state of Mato Grosso
(MT), Brazil.The study was conducted along the Fish river, located
on the road linking Cuiabáto Chapada dos Guimarães,MT. CDC
(Center for Disease Control) light traps where installed close to four
residences at 18 PM and collected at 6 AM of the next day between
June and November, 2013.Captured insects where stored in ethanol
70% and identified using Young and Duncan classification at the
Entomology Laboratory. In this study, 29 Lutzomyia sandflies
belonging to 10 species were captured. The most abundant was
Lutzomyia cruzi (11/29; 37.93%), followed by Lutzomyia whitmani
and Lutzomyia acanthopharyns (4/29;13.79% of each), Lutzomyia
evandroi (3/29;10.34%), Lutzomyia longipalpis (2/29; 6.90%) and
Lutzomyia punctigeniculata, Lutzomyia corumbaensis, Lutzomyia
lenti, Lutzomyia lutiziana and Lutzomyia sp. (1/29; 3.45% of
each).August and September were the months with the highest
occurrence of sandflies. Lutzomyia whitmani (main vector of
American Cutaneous Leishmaniasis in MT), Lutzomyia cruzi and
Lutzomyia longipalpis (vector species of visceral leishmaniasis) were
the most frequent sandflies found. This area will remain under
entomological and epidemiological surveillance, considering the risk
of cutaneous and visceral leishmaniasis transmission.
Leishmaniasis is a group of diseases caused by protozoan of the genus
Leishmania, which can be visceral or cutaneous manifestations that is
caused by a protozoan and transmitted by an insect of the genus
Lutzomyia, called sandflies. This study aims to monitor the presence
of sandflies in the UFMT Zoological Park, for the control of vectors
and adopt preventive measures in the transmission of leishmaniasis in
animals and humans. Catches were made with CDC light trap in four
points as farmed birds, small mammals, large mammals and reptiles.
Samples were collected monthly for three consecutive nights from
January 2012 to December 2013. 121 individuals of the genera
Brumptomyia = 7 (5.8%) and Lutzomyia carmelinoi = 3 (2.5%) were
captured; L. cruzi = 76 (62.8%); L. hermanalenti = 5 (4.1%); L. lenti =
7 (5.8%) L. whitmani = 20 (16.5%) and Lutzomyia sp = 3 (2.5%).The
sex ratio was 67.8% males and 32.2% females. The months of highest
incidence were June 2012 and 2013. Among the collected species L.
cruzi is considered vector of visceral leishmaniasis and L. whitmani of
American cutaneous leishmaniasis. The results indicate that the
strategy of control and monitoring of the vectors must be continuous
throughout the year.
N° 26 (1) 2015
P O P U L AT I O N D Y N A M I C S A N D H A B I TAT
PREFERENCES OF PHLEBOTOMUS ORIENTALIS IN
EXTRA-DOMESTIC HABITATS OF METEMA HUMERA LOWLANDS - KALA-AZAR ENDEMIC
AREAS IN NORTHWEST ETHIOPIA
ECOLOGY OF LUTZOMYIA LONGIPALPIS IN AN
AREA OF VISCERAL LEISHMANIASIS
TRANSMISSION IN NORTH-EASTERN BRAZIL
Wossenseged Lemma1*, Habte Tekie2 Meshesha Balkew3, GebreMichael3, Alon Warburg4, Asrat Hailu5
74
Pietra Lemos Costa, Filipe Dantas-Torres, Fernando José da Silva,
Vanessa Cristina Fitipaldi Veloso Guimarães, Kamila Gaudêncio, Sinval
Pinto Brandão Filho
Centro de Pesquisas Aggeu Magalhães
1
Department of Parasitology, School of Biomedical and Laboratory Sciences, College
of Medicine and Health Sciences, University of Gondar, Gondar, Ethiopia. 2Department
of zoology, Faculty of Life Science, Addis Ababa University, Addis Ababa, Ethiopia.
3
Aklilu Lemma Institute of Pathobiology, Addis Ababa University, Addis Ababa,
Ethiopia. 4Department of Microbiology & Molecular Genetics, The Institute for
Medical Research Israel-Canada, The Kuvin Centre for the Study of Infectious &
University of Jerusalem, 91120, Israel. 5Department of Microbiology, Immunology &
Parasitology, Faculty of Medicine, Addis Ababa University, Addis Ababa, Ethiopia
Keywords: Population dynamics; Phlebotomus orientalis
Abstract Background. Metema-Humera lowlands are endemic for
kala-azar (visceral leishmaniasis). These lowlands are characterized
by black clay soil and used for growing sesame, sorghum and cotton
for commercial purposes. Objectives. The aim of this study was to
study seasonal dynamics and habitat preference of Phlebotomus
orientalis in extra-domestic habitats. Methods. CDC-light trap [CDCLT] and Sticky Trap [ST] were used to collect sand flies from various
habitats before species identification by their morphological
characteristics using appropriate keys. Data summarized and analyzed
included: species, sex, density, habitats, type of trap used and date
(month). Results. A total of 389,207 sand flies using CDC-LT (n=992)
and ST (n=5730) were collected from May 17, 2011 to June 6, 2012.
The highest Mean Monthly Density (MMD) of P. orientalis trapped by
CDC-LT was found in thickets of Acacia seyal in March (MMD of
64.11 + 75.87). The corresponding highest MMD of P. orientalis
trapped by STs was found in April (58.69 + 85.20) in agricultural field.
No P. orientalis were caught in September using CDC traps and July October using sticky traps. The overall MMD of P. orientalis trapped
by CDC-LT was 14. 83 + 28.03 in agricultural field, 19.37 + 36.42 in
thickets of A. seyal, and 3.55 + 6.082 in dense mixed forest. Localitywise, the MMD of P. orientalis trapped by CDC-LT did not show
significant statistical difference (p=0.135). Conclusions. Agricultural
fields and thickets of A. seyal, habitats which exhibit extensive soil
cracks and fissures, as opposed to dense mixed forests, serve as
preferred breeding sites for P. orientalis.
lemos.pietra@gmail.com
Keywords: Sand flies, Phlebotominae, Lutzomyia longipalpis,
Ecology
Visceral leishmaniasis is a major public health issue in South
America, where the disease is rapidly spreading. Changes in ecology
and distribution of the principal vector, Lutzomyia longipalpis are
among the factors accounting for the increasing incidence of the
disease in this region. However, information about the ecology of L.
longipalpis is still incipient, which may directly impair the
implementation of effective control programs. Herein, the ecology of
L. longipalpis was studied in a focus of visceral leishmaniasis in
north-eastern Brazil. From August 2009 to August 2010,
phlebotomine sand flies were monthly collected in four localities
using CDC light traps (∼37 per month) and a lantern-baited Shannon
trap with mouth aspirators. A total of 24,226 phlebotomine sand flies
were collected with light traps and 375 with mouth aspirators. The
most abundant species was L. longipalpis, representing 97.9% of the
specimens collected with light traps and 91.5% with the mouth
aspirator. Other species (Lutzomyia evandroi, Lutzomyia lenti and
Lutzomyia sallesi) were found in low numbers. Most phlebotomine
sand flies (94.6%) were collected at chicken coops and corrals. No
significant correlation was found between the monthly abundance of
phlebotomine sand flies and the monthly averages of temperature,
relative humidity or rainfall. However, interestingly enough, 82.4%
of L. longipalpis specimens were collected in months when relative
humidity surpassed 75%. This study points out that this vector is well
adapted to live in different habitats and to different climate
conditions. It also suggests that some north-eastern populations of L.
longipalpis may be more xerotolerant than southern populations.
Further studies to assess the relationship between microclimate and L.
longipalpis density in different Brazilian regions are advised.
N° 26 (1) 2015
75
DIVERSITY OF PHLEBOTOMINE SAND FLIES IN AN
AMERICAN CUTANEOUS LEISHAMANIASIS
ENDEMIC AREA IN NORTH-EASTERN BRAZIL
A REAL-TIME PCR FOR CHARACTERIZATION OF
M U LT I P L E B L O O D S O U R C E S I N F I E L D COLLECTED PHLEBOTOMINE SAND FLIES
Débora Elienai de Oliveira Miranda, Kamila Gaudêncio da Silva, Maria
Aparecida da Gloria Faustino, Leucio Câmara Alves, Sinval Pinto
Brandão-Filho, Filipe Dantas-Torres, Gílcia Aparecida de Carvalho
Kamila Gaudêncio da Silva, Rayana Carla Silva de Morais, Débora
Elienai de Oliveira Miranda, Abilene Cristina de Arruda Moura, Sinval
Pinto Brandão-Filho, Milena de Paiva Cavalcanti, Filipe Dantas-Torres
Department of Veterinary Medicine, Federal Rural University of Pernambuco, 52171900, Recife, Pernambuco, Brazil, Department de Immunology, Aggeu Magalhães
Research Centre, 50670-420 Recife, Pernambuco, Brazil, Department of Veterinary
Medicine, Università degli Studi di Bari, 70010 Valenzano, Bari, Italy, Academic Unit
of Garanhuns, Federal Rural University of Pernambuco, Garanhuns, Pernambuco,
Brazil, 55296-901.
Departamento de Imunologia, Centro de Pesquisas Aggeu Magalhães, 50670-420
Recife, Pernambuco, Brazil, Dipartimento di Medicina Veterinaria, Università degli
Studi di Bari, 70010 Valenzano, Bari, Italy.
debora-myranda@hotmail.com
Keywords: Sand flies, source blood and qPCR
Keywords: Ecology, phlebotomine and vectors
Zoonotic leishmaniasis caused by Leishmania infantum is endemic in
the international touristic Algarve region (AR) in southern Portugal
being Phlebotomus perniciosus the only proven vector up to now.
However, L. major DNA was recently detected in one Sergentomyia
minuta specimen collected in this region. Nevertheless, and albeit up
to now no autochthonous cutaneous leishmaniasis due to L. major or
L. tropica has been detected in Portugal, their vectors (i.e. P. papatasi
and P. sergenti, respectively) were also identified. We report a three
years survey carried out to update the distribution, density,
Leishmania infection rate and to identify blood-meal sources of
phlebotomine sand flies in AR. From May to October 2011, 2012 and
2013, CDC light traps and sticky papers were set up in 11 sampling
points during 3 consecutive days per month. Collecting places
included domestic, peri-domestic and sylvatic environments. A total
of 4961 sand flies (2569 males and 2392 females) were collected and
identified morphologically. At least one specimen of the five known
species present in Portugal (P. ariasi, P. papatasi, P. pernicious, P.
sergenti and S. minuta) was captured. The predominant species was S.
minuta followed by P. perniciosus highlighting the high plasticity of
both species to adapt to different environments. In addition, blood
meals revealed that P. perniciosus and S. minuta fed on a wide range
of hosts (i.e. humans, horses, cattle, sheep, pigs, poultry, rabbits and
lizards). Although Sergentomyia genus is known as a vector of
Sauroleishmania sp. its epidemiological significance should not be
neglected as they have already been suspected to be involved in L.
donovani and L. major transmission among humans in India and Mali,
respectively and L. infantum among dogs in Senegal. Our results
reinforce the importance of systematic surveillance with extensive
epidemiological surveys on Leishmania hosts and vectors to
investigate the transmission, distribution and spreading of
Leishmania sp. infection in order to detect areas of higher of risk of
vector transmission and to implement specific control measures.
Funding: This work was partially supported by EDENext (FP7INCO-CT-2010-261504). C. Maia (SFRH/BPD/44082/2008) holds a
fellowship from Fundação para a Ciência e a Tecnologia, Ministério
da Ciência, Tecnologia e Ensino Superior, Portugal.
Phlebotomine sand flies are vectors of protozoa of the genus
Leishmania, the causative agents of leishmaniasis. The ecology of
phlebotomine sand flies is of research interest because it can provide
background information for a better understanding of the
epidemiology of leishmaniasis and to predict the spatial and temporal
variations in the risk of infection in a given area. American cutaneous
leishmaniasis (ACL) is among the most important tropical diseases in
Brazil and in several developing countries in the Neotropics. The
present study aimed to identify the different species that make up the
phlebotomine sand flies fauna in an area of the municipality of the
Ipojuca, Pernambuco state, Brazil. Phlebotomine sand fly collections
were conducted from August 2013 to February 2014, using 7 CDC
light traps for three consecutive nights from 17:00 pm to 6:00 am, on a
monthly basis. Each trap was placed at 1.5 m above the ground, being
installed in three different environments: indoors, peridomicile (i.e.,
forest edge) and in the forest. The capture sites were usually near the
burrows of wild animals, trunks of large trees, in shady and moisturerich environments. The captured specimens were examined,
separated according to sex and kept in 70% ethanol. The sandflies
were mounted on slides and identified morphologically. A total of
2,889 specimens belonging to 17 species were identified, namely: L.
choti (n = 1,680), L. whitmani (n = 495), L. quinquefer (n = 223), L.
sordellii (n = 209), L. longispina (n = 84), L. amazonensis (n = 52), L.
evandroi (n = 49), L. brasiliensis (n = 28), L. naftalekatzi (n = 27), L.
barrettoi barrettoi (n = 17), L. capixaba (n = 10), L. shannoni (n = 4),
L. aragaoi (n = 2) L. complexa (n = 5) L. furcata (n = 2), L. wellcomei
(n = 1) and L. ayrozai (n = 1). Males (n = 1,431) predominated over
females (n = 1,458). L. choti and L. whitmani were the most abundant
species, comprising together 75.2% of the specimens collected. The
study demonstrates an important variety of phlebotomine sand fly
species in the studied area and that some of them may be associated
with human cases of ACL. The finding of L. whitmani, a proven vector
of Leishmania braziliensis in several Brazilian regions indicates its
probable involvement in the transmission of ACL in this municipality.
N° 26 (1) 2015
76
A THREE CONSECUTIVE YEARS PHLEBOTOMINE
SAND FLY SURVEY IN A ZOONOTIC LEISHMANIASIS
ENDEMIC FOCUS, THE ALGARVE REGION,
PORTUGAL
NOCTURNAL ACTIVITIES AND HOST
PREFERENCES OF P. ORIENTALIS IN EXTRADOMESTIC HABITATS OF KALA-AZAR ENDEMIC
METEMA–HUMERA LOWLANDS, NORTH WEST
ETHIOPIA
C.Maia1,2,3, J. Cristóvão 1,2, F. B. Freitas1, M.O. Afonso1,4, L. Campino1,5
1
Unidade de Parasitologia Médica, Instituto de Higiene e Medicina Tropical,
Universidade Nova de Lisboa, Lisboa, Portugal 2Centro de Malária e outras Doenças
Tropicais 3Faculdade de Medicina Veterinária, Universidade Lusófona de
Humanidades e Tecnologias, Lisboa, Portugal 4Unidade de Parasitologia e
Microbiologia Médicas 5Departamento de Ciências Biomédicas e Medicina,
Universidade do Algarve, Faro, Portugal
carlamaia@ihmt.unl.pt
Keywords: Phlebotomine sand fly survey; zoonotic leishmaniasis;
Algarve; Portugal
Zoonotic leishmaniasis caused by Leishmania infantum is endemic in
the international touristic Algarve region (AR) in southern Portugal
being Phlebotomus perniciosus the only proven vector up to now.
However, L. major DNA was recently detected in one Sergentomyia
minuta specimen collected in this region. Nevertheless, and albeit up
to now no autochthonous cutaneous leishmaniasis due to L. major or
L. tropica has been detected in Portugal, their vectors (i.e. P. papatasi
and P. sergenti, respectively) were also identified. We report a three
years survey carried out to update the distribution, density,
Leishmania infection rate and to identify blood-meal sources of
phlebotomine sand flies in AR. From May to October 2011, 2012 and
2013, CDC light traps and sticky papers were set up in 11 sampling
points during 3 consecutive days per month. Collecting places
included domestic, peri-domestic and sylvatic environments. A total
of 4961 sand flies (2569 males and 2392 females) were collected and
identified morphologically. At least one specimen of the five known
species present in Portugal (P. ariasi, P. papatasi, P. pernicious, P.
sergenti and S. minuta) was captured. The predominant species was S.
minuta followed by P. perniciosus highlighting the high plasticity of
both species to adapt to different environments. In addition, blood
meals revealed that P. perniciosus and S. minuta fed on a wide range of
hosts (i.e. humans, horses, cattle, sheep, pigs, poultry, rabbits and
lizards). Although Sergentomyia genus is known as a vector of
Sauroleishmania sp. its epidemiological significance should not be
neglected as they have already been suspected to be involved in L.
donovani and L. major transmission among humans in India and Mali,
respectively and L. infantum among dogs in Senegal. Our results
reinforce the importance of systematic surveillance with extensive
epidemiological surveys on Leishmania hosts and vectors to
investigate the transmission, distribution and spreading of Leishmania
sp. infection in order to detect areas of higher of risk of vector
transmission and to implement specific control measures. Funding:
This work was partially supported by EDENext (FP7-INCO-CT2010-261504). C. Maia (SFRH/BPD/44082/2008) holds a fellowship
from Fundação para a Ciência e a Tecnologia, Ministério da Ciência,
Tecnologia e Ensino Superior, Portugal.
Wossenseged Lemma1, Habte Tekie2 , Ibrahim Abassi3, Meshesha
Balkew3, Teshome Gebre-Michael3, Alon Warburg3 ,Asrat Hailu5
1
*Department of Parasitology, School of Biomedical and Laboratory Sciences, College
of Medicine and Health Sciences, University of Gondar, Gondar, Ethiopia.
2
Department of zoology, Faculty of Life Science, Addis Ababa University, Addis Ababa,
Ethiopia. 3Department of Microbiology & Molecular Genetics, The Institute for
Medical Research Israel-Canada, The Kuvin Centre for the Study of Infectious &
University of Jerusalem, 91120, Israel. 3Aklilu Lemma Institute of Pathobiology, Addis
Ababa University, Addis Ababa, Ethiopia. 5Department of Microbiology, Immunology
& Parasitology, Faculty of Medicine, Addis Ababa University, Addis Ababa, Ethiopia
Keywords: Nocturnal Activities, Phlebotomus orientalis
Background: P. orientalis takes blood from a variety of wild and
domestic animals and transmit L. donovani from unknown reservoir
hosts to human in extra-domestic habitats in Metema - Humera
lowlands. Objective: The aim of this study was to determine the
nocturnal activities of P. orientalis and their preferred host for blood
meal during night time. Methods: Hourly collections of P. orientalis
using CDC traps from farm, near goat and sheep night shelter and
thickets of Acacia seyal were used for the study nocturnal activities of
P. orientalis. Host preference study of P. orientalis towards rodents
was studied using Turner's box trap. Self bait technique was used to
study the host preference of P. orientalis towards human in addition
to blood meal analysis of this vector using Reverse Line Blot (RLB) of
cytochrome b PCR product. Result. A total of 6083 P. orientalis (2702
male and 3381 female) were collected at one hour interval from
farms near camps, sparse thickets of Acacia seyal and near animal
shelters (sheep and goat) using 22 CDC traps during January,
February March and May, 2013. Nocturnal activities of female P.
oriental were different at different hours of the night in the farm
(p=0.023) and near animal shelter (p=0.00). However, the activities
were the same for the different hours of the night in the thickets of
Acacia seyal (p=0.144). The peaks activities of P. oriental were at
1.00 a. m near animal shelter (134.0 ±7.21) compared to 3.00 a. m
(66.33 ± 46.40) in the farm. A total of 984 female P. orientalis from
farm near the camp and animal house were collected at one hour
intervals using self bait. The highest mean landing of P. orientalis
were 38.50 ± 31.88 at 12 pm with maximum number of 86 with no
statistical difference for mean number of P. orientalis biting man (p
= 0.066). P. orientalis were attracted more to human than sheep and
goats ((p=0.000). P. orientalis were not attracted to the different
species of rodents. Blood meal analysis of the 25 blood fed P.
orientalis using cyt b PCR and RLB captured 7 human(28%), 9
mixed (human and cattle) (36%), 2 cattle(8%). There were samples
(28%) with bands on agarose gel but not in nitrocellulose membrane
indicating the existence of other animals outside the 11 probes used.
Conclusion: P. orientalis feed on goats, sheep, cattle and human
throughout the night with peak biting after mid night.
N° 26 (1) 2015
77
PREVALENCE OF LEISHMANIA SPP. INFECTION
AND HOST - BLOOD FEEDING OF SANDFLIES
TRANSMITTERS OF CUTANEOUS LEISHMANIASIS
IN AN ENDEMIC LOCALITY IN THE DISTRICT OF
CAPIRA, PANAMA
A STUDY ON NOCTURNAL ACTIVITY OF SAND
FLIES IN A LEISHMANIASIS ENDEMIC VILLAGE
LOCATED IN AYDIN PROVINCE OF TURKEY
Suha K. Arserim1, Mehmet Karakus2, Yusuf Ozbel3
Chystrie Rigg1,2, Anayansi Valderrama3,
Chavez4 y Azael Saldaña1
José E. Calzada1,
Luis F.
1
Celal Bayar University, Vocational School of Health Services, Manisa, TURKEY; 2Ege
University Institute of Health Sciences, Bornova, Izmir, TURKEY; 3Ege University
Faculty of Medicine, Department of Parasitology, Bornova, Izmir, Turkey
1
Department of Parasitology, Gorgas Memorial Institute for Health Studies, Panama;
Central American Program of Masters in Entomology, University of Panama,
Panama; 3Department of Medical Entomology, Gorgas Memorial Institute for Health
Studies, Panama; 4Institute of Tropical Medicine (NEKKEN), Nagasaki University,
Japan.
2
suhaarserim@hotmail.com
Keywords: Sand fly, nocturnal activity, Turkey
chrigg@gorgas.gob.pa
Keywords: Leishmaniasis, infection, Blood Feeding, Panama.
The community of Trinidad de las Minas in the District of Capira,
Province of Panama, is located in a region where an intense
transmission of cutaneous leishmaniasis (CL) have been reported. In
this area, the increase of CL cases ranges between 137 to 508 per year,
with an incidence that corresponds to 20% of the whole country. The
purpose of this work is to demonstrate in this community, the
prevalence of Leishmania spp. infection and host-blood feeding in
anthropophilic and zoophilic sandflies, before and after vector control
intervention with insecticide thermal fogging (ITF). From April 2010
to June 2011, sandflies where collected using CDC light traps, inside
(domicile) and around (peridomicile) of 24 houses. To evaluate the
Leishmania infection, we analyzed the DNA sand fly pools (main
species, 5-10 individuals) by ITS-1 gene PCR/sequencing. Positive
pools were analyzed by cytochrome b PCR/sequencing, to
differentiate the host-blood feeding patterns (domestic, wild
mammals or birds). Of 455 analyzed pools, 253 resulted positives for
PCR-ITS1. Of them, 164 correspond to important leishmaniasis
vectors found in this community: Lutzomyia trapidoi, Lu. gomezi and
Lu. panamensis. The estimated prevalence to 95% depending on the
algorithms of the epitool tool (pooled prevalence for variable pool size
and perfect tests) was 0.09 to 0.11 before and after the intervention
respectively. In the detection of host-blood feeding, it was found that
birds and pigs were the main sources of blood, followed by humans
and dogs. These findings demonstrate the dynamics of Leishmania
infection and blood feeding in vector sandflies, before and after an
ITF, a method often used to control mosquitoes and other arthropods
in Panama. The results confirm the infection of the main species of
anthropophilic sandflies with Leishmania spp. in this hyperendemic
focus of CL. A close association between these vector species, humans
and some domestic animals was also supported, emphasizing the
importance of implementing active surveillance using these
parameters to predict outbreaks and develop new control strategies or
integrate existing ones to reduce cutaneous leishmaniasis in this
region.
The nocturnal activity of sand flies in an endemic focus for canine and
cutaneous leishmaniasis in western part of Turkey was studied using
CDC light traps. The traps were set up in different micro-localities,
such as human house, cattle shed, chicken coop and sheep area within
60-70 meters in diameter. In the study night, a total of 263 sand flies,
six species belonging to Phlebotomus and two species belonging to
Sergentomyia genera, were caught. P. tobbi was found to be abundant
species (60.46%) while P. neglectus/syriacus was second one
(21.29%). The activity of sand flies started at 21:00-22:00h, reached
maximum between 01:00 and 03:00h and steadily decreased
thereafter and finished at 05:00h in all micro-localities. Female sand
flies was always 2-3 times in high number than males. The study was
showed the highest activity of probable vector species, P. tobbi, in the
area and proved the importance of the studies on seasonal and daily
activity of sand flies in endemic areas. This study was partially funded
by EU grant GOCE-2003-010284 EDENext.
N° 26 (1) 2015
78
SEASONAL ACTIVITIES AND ECOLOGICAL
F E AT U R E S O F S A N D F L I E S ( D I P T E R A :
PSYCHODIDAE: PHLEBOTOMINAE) IN AYDIN
MOUNTAINS AND SURROUNDINGS LOCATED IN
W E S T E R N T U R K E Y, E N D E M I C A R E A F O R
CUTANEOUS LEISHMANIASIS
BIONOMY OF PHLEBOTOMINE SANDFLIES
(PHLEBOTOMINAE) IN AN OUTBREAK AREA OF
VISCERAL LEISHMANIASIS IN CORRIENTES,
ARGENTINA
Araujo AV 1, Benítez Ibalo AP 1, Rosa JR 2; Oscherov EB 1; Salomón OD 3
Suha K. Arserim1, Ahmet Mermer2, I. Cuneyt Balcioglu3, V. Ozge Ermis4;
Yusuf Ozbel5
1
Celal Bayar University Vocational School of Health Services, Manisa, Turkey; 2Ege
University, Science Faculty, Department of Zoology, Bornova, Izmir, Turkey; 3Celal
Bayar University Faculty of Medicine, Department of Parasitology, Manisa, Turkey;
4
Celal Bayar University Institute of Science, Manisa, Turkey; 5Ege University Faculty of
Medicine, Department of Parasitology, Izmir, Turkey
1
Lab. Biología de los Parásitos. Facultad de Ciencias Exactas y Naturales y
Agrimensura. Universidad Nacional del Nordeste (UNNE). Red de Investigación de la
Leishmaniasis en Argentina (REDILA). 2 Lab. Entomología. Instituto de Medicina
Regional. UNNE. REDILA. 3 Instituto de Medicina Tropical (InMET), Puerto Iguazú,
Misiones, Argentina. REDILA.
analiav.araujo@gmail.com
suhaarserim@hotmail.com
Keywords: Leishmania , Epidemiology, Lutzomyia
Keywords: Sand fly, Seasonal activity, Leishmaniasis, Aydın
Mountains
The abundance study in time and space of Phlebotominae through the
area of the City of Corrientes (27°29′S;58°49′W) was based in human
and canine cases with Visceral Leishmaniasis (VL) reported in the
province and the previous records of the urban vector Lutzomyia
longipalpis in the city. This preliminary study involves collection
performed in three sites according to its transitional environments:
periurban-rural (ENU), periurban-urban (EPU) and urban-centric
(EU). In each environment three households were selected based on
the ´worst scenario´ criteria. From March to December 2013 the
captures were made fortnightly during three consecutive days using
CDC type-light trap in each peridomiciliary habitat, from 17:00hs to
8:00hs. In total 449 Phlebotominae were captured: Lu. longipalpis
(51.2%), Nyssomyia neivai (43.7%), Migonemyia migonei (2.2%),
Micropygomia quinquefer (2.0%) and Evandromyia females from
cortelezzii complex (0.9%). The sandfly maximal abundance was in
March (32.7%) with an average temperature of 22ºC, and minimum
abundance during August (0.2%) with temperatures of approximately
14,3ºC. Lu. longipalpis was recorded in all the seasons with the
highest abundance in March and April (n=119) during autumn, and
the lowest abundance (n=27) in winter, with a progressive increase of
the abundance in the captures from September, at the beginning of the
spring. The main abundance of Ny. neivai was also in autumn, but its
captures increased since October. Although Lu. longipalpis was
recorded in the three environments, was more abundant in EPU and
EU, while Ny. neivai was more abundant in ENU. Mi. migonei was
recorded in autumn and spring in ENU and EPU but due to the low
number collected may be considered as an occasional specie. The
consistent presence of Lu. longipalpis in time and space in the urban
area and the occasional captures of Mi. migonei, main and putative
vectors of Leishmania infantum respectively, besides the reports of
autochthonous cases of human and canine VL, suggests a risk of
transmission along the year. However, this risk is highly
heterogeneous in time and space, lower in winter despite of the fact
that the actual incidence of cases could increase early in this season
due to the abundance vectors peaked in autumn and the parasite
intrinsic incubation time. Financial support: Proyecto (F008/09) Res.
Nº 1080/09. Periodo 2010-2013. UNNE. PICTO-ANLIS 2011.
The aim of this study was to determine (a) the seasonal activities and
ecological features of sand fly population, (b) identifying sand fly
fauna, (c) preparing maps for showing the distribution of existing
species, and (d) analyzing the diversity and population sizes of sand
fly species in The Aydın Mountains and surroundings located in
western Turkey, an important endemic focus for cutaneous and canine
leishmaniasis. During the entomological survey 10 locations were
selected according to the altitude and sand flies were collected using
CDC light traps and sticky paper traps from June 2009 to July 2010 by
15-day intervals in summer season and 30-day intervals in winter
season. Temperature, humidity and general environmental parameters
of study area were recorded using data loggers. Totally, 6817 sand flies
were collected and dissected. The identification was made according
to the Mediterranean keys. In total, 12 species of sand flies were
identified, which are 9 species belonging to genus Phlebotomus (P.
papatasi 5.79%, P. similis 4.44%, P. alexandri 4.20%, P. mascittii
0.28%, P. neglectus/syriacus 29.91%, P. burneyi 0.62%, P. tobbi
22.58%, P. simici 3.45% and P. brevis 0.44%) and 3 species are
belonging to genus Sergentomyia (S. dentata 22.81%, S. antennata
1.55% and S. minuta 2.39%). In conclusion, 12 sand fly species are
detected in the study area and maps were produced. P. neglectus, P.
tobbi and P. similis, the well known vector species for Leishmania
infantum and L. tropica were found dominant species by several
analyses, respectively. The activity period was determined as between
April and October, for 7 months. Population abundance of P. similis
was the highest at the altitude of 400-600m in July while P. neglectus
and P. tobbi were found more abundant at the altitude of 200-400m in
September. These results on sand fly ecology area could be useful for
establishing or improving effective control programs.
N° 26 (1) 2015
79
ATTRACTION OF ETHIOPIAN PHLEBOTOMINE
SAND FLIES (DIPTERA: PSYCHODIDAE) TO LIGHT
AND SUGAR-YEAST MIXTURES (CO2)
ECOLOGICAL NICHE MODELING PREDICTS
S O U T H WA R D E X PA N S I O N O F L U T Z O M Y I A
F L AV I S C U T E L L ATA , V E C TO R O F D I F F U S E
CUTANEOUS LEISHMANIASIS IN SOUTH AMERICA,
UNDER CLIMATE CHANGE
Oscar David Kirstein, Roy Faiman, Araya Gebreselassie, Asrat Hailu,
Teshome Gebre-Michael and Alon Warburg
Department of Microbiology and Molecular Genetics, The Institute for Medical
Research Israel-Canada, The Kuvin Centre for the Study of Infectious and Tropical
Diseases, The Hebrew University – Hadassah Medical School, The Hebrew University
of Jerusalem, Israel.
od.kirstein@mail.huji.ac.il
Keywords: Attractants, Phlebotomus orientalis, Visceral
leishmaniasis, Yeast fermentation.
Visceral leishmaniasis (VL) known as Kala-Azar is a serious systemic
disease caused by Leishmania donovani parasites
(Trypanosomatidae: Kinetoplastida). The disease is prevalent in the
Indian Sub-continent, East Africa and Brazil. In Africa, the worst
affected regions are in Sudan, with an estimated 15,000-20,000 cases
annually and, Ethiopia with 5,000-7,000 cases a year. The main vector
of VL in Sudan and Northern Ethiopia is Phlebotomus orientalis, a
sand fly frequently found in association with Acacia spp and Balanite
spp woodlands. Methods: To optimize sampling of sand flies for
epidemiological studies in remote areas we tested different means of
attraction. Miniature suction traps employing 2AA batteries (3 V)
were deployed in the up-draft orientation and baited with chemical
light-sticks (Red, Yellow and Green), or bakers' yeast in sugar solution
(emitting CO2 and perhaps other attractants). These traps were
compared with standard CDC incandescent light traps employing 6 V
20 batteries. Trials were conducted during two consecutive years at
different localities around Sheraro, a town in West Tigray, Northern
Ethiopia.Results: The sand fly species composition was similar but
not identical in the different locations tested with different
Sergentomyia spp. predominating. Phlebotomus spp. comprised less
than 1% of the catches during the first year trials (November –
December 2011) but increased markedly during the second year trials
performed later in the dry season at the height of the sand fly season in
February 2012. Although there did not appear to be a species bias
towards different light wave-lengths, fermenting yeast in sugar
solution attracted relatively more Phlebotomus spp. and Sergentomyia
schwetzi.Conclusions: Although the standard 6 V CDC incandescent
light traps captured more sand flies, light-weight (~350 g) 3 V suction
traps baited with chemical light-sticks were shown to be effective
means of monitoring sand flies. Such traps operated without light and
baited with yeast in sugar solution caught relatively more
Phlebotomus spp.
Bruno Moreira de Carvalho1,2,3, Mariana Moncassim Vale2, Maria Lucia
Lorini4, Margarete Martins dos Santos Afonso1, Elizabeth Ferreira
Rangel3
1
Programa de Pós-Graduação em Ecologia e Evolução, Universidade do Estado do
Rio de Janeiro; 2Departamento de Ecologia, Instituto de Biologia, Universidade
Federal do Rio de Janeiro; 3Laboratório de Transmissores de Leishmanioses, Instituto
Oswaldo Cruz, Fundação Oswaldo Cruz; 4Departamento de Ciências Naturais,
Instituto de Biociências, Universidade Federal do Estado do Rio de Janeiro.
brunomc@ioc.fiocruz.br
Keywords: Ecological niche modeling, climate change, Lutzomyia
flaviscutellata
According to the Fifth Assessment Report of the Intergovernmental
Panel on Climate Change, climate change will exacerbate health
problems that already exist. Leishmaniasis is a climate-sensitive
disease, occupying a characteristic climate space that is strongly
affected by changes in rainfall and temperature. Climate change,
therefore, should alter the distribution of leishmaniasis vectors. We
modeled the climatic suitability of Lutzomyia flaviscutellata, main
vector of diffuse cutaneous leishmaniasis (DCL) in South America,
and projected the models under climate change scenarios to assess
possible range changes. To provide occurrence data, a database of 328
records of L. flaviscutellata was compiled from the literature,
museum collections and Brazilian Health Departments. Six
bioclimatic variables derived from temperature and precipitation
were used as predictors. Models were produced based on eight
different algorithms – Bioclim, Domain, Mahalanobis Distance,
Generalized Linear Model, Generalized Additive Model, Maxent,
Random Forest and Support Vector Machines – and evaluated by a
threshold-independent method (AUC). Ensemble models were
produced by overlapping binary predictions of each algorithm. Future
projections used 15 most recent climate models for 2050 (CMIP5), in
climate change scenarios representing two greenhouse gas
concentration pathways: stabilization (RCP4.5) and high increase
(RCP8.5). Single-models produced by each algorithm and climate
model showed fair to good performance (0.71<AUC<0.87), but
resulted in quite dissimilar predictions. Compiled records of
Lutzomyia flaviscutellata are located in Bolivia, Brazil, Colombia,
Ecuador, French Guiana, Peru, Suriname, Trinidad and Tobago and
Venezuela. Currently, the species is widely distributed in the Amazon,
with some records in Atlantic Forest and savannah regions to the
south. Most consensus models agree that the area of climatic
suitability of L. flaviscutellata should expand in both climate change
scenarios, and all consensus models agree that the vector should
experience improved climatic suitability towards the south. Human
cases of DCL in southeast Brazil are currently rare, although the
disease seems to be gradually expanding its occurrence southward. If
the vector reaches these climatically suitable areas and its dispersion
is followed by competent hosts and parasites, these can become DCL
risk areas, especially because these are the most populated areas
within the species' range.
N° 26 (1) 2015
IDENTIFICATION OF PLANTS FED UPON BY
PHLEBOTOMINE SAND FLIES IN ETHIOPIA
H A B I TAT S U I TA B I L I T Y F O R L U T Z O M Y I A
(NYSSOMYIA) WHITMANI AND AMERICAN
CUTANEOUS LEISHMANIASIS (ACL) IN BRAZIL
Oscar David Kirstein; Ibrahim Abassi; Araya Gebreselassie; Teshome
Gebre-Michael; Asrat Hailu and Alon Warburg
Department of Microbiology and Molecular Genetics, The Institute for Medical
Research Israel-Canada, The Kuvin Centre for the Study of Infectious and Tropical
Diseases, The Hebrew University – Hadassah Medical School, The Hebrew University
of Jerusalem, Jerusalem - Israel
od.kirstein@mail.huji.ac.il
Keywords: PCR, Plant DNA, Sand fly, Ethiopia
Only the female sand flies transmit Leishmania parasites when they
suck blood required for maturation of the eggs. However, both sexes
feed on plant-derived sugars by imbibing plant sap, nectar and/or
honeydew. Therefore, determining the sources of sand fly sugar
meals can improve our understanding of the ecology and transmission
of leishmaniasis. The attraction of sand flies to certain habitats
probably depends on the vegetation and their preference for certain
plants as food sources, resting places or even breeding habitats. Thus,
certain plant communities may comprise risk factors for contracting
leishmaniasis. For example, the sand fly Phlebotomus orientalis,
vector of visceral leishmaniasis (VL) in Sudan and Northern Ethiopia
is frequently associated with wooded areas dominated by Acacia sp
and Balanites sp. In fact, Acacia forests are commonly perceived as an
important factor in contracting Kala Azar in these areas. A botanical
survey was conducted in different areas around Sheraro town, in
Tigray, northern Ethiopia. Twenty four of the plant species
predominating during the dry season were collected and identified
based on classical and molecular plant taxonomy. For molecular
identification, PCR systems were developed for three chloroplast
genes rbcl, MatK, ATP synthase and the nuclear ribosomal gene
(ITS1). Plant specific primers were designed after DNA alignment of
DNA sequences and primer annealing sites were selected in the
conserved sequences. The PCR amplified regions were sufficiently
variable to identify the different species by DNA sequencing. The two
primer pairs for rbcl and ATP synthase were shown to be the most
suitable. In preliminary experiments using the new PCR system, we
successfully amplified plant DNA from Ph. orientalis sand flies
collected in different ecotopes. Further studies are ongoing to adapt a
rapid and inexpensive DNA identification protocol using reverse
blotting in order to replace the more costly time-consuming DNA
sequencing.
80
Simone M. da Costaa, José L. P. Cordeirob, Margarete M.S. Afonsoa,
Elizabeth F. Rangela
Laboratório de Transmissores de Leishmanioses, IOC/FIOCRUZ, Av. Brasil n° 4365,
21040360, RJ, Brazil; bCampus Fiocruz da Mata Atlântica, CFMA/Presidência,
FIOCRUZ, RJ, Brazil.
scosta80@hotmail.com
Keywords: Lutzomyia (Nyssomyia) whitmani, Leishmaniose
Tegumentar Americana, alterações ambientais
The impact of environmental change as well as those determined by
human activity, combined with different aspects of the vector,
parasite, and reservoir host ecology, play an important role in the
dissemination of ACL in Brazil. In this context highlight L. (N.)
whitmani, most important vector of L. (V.) braziliensis and L. (V.)
shawi. The characterization of factors that influence the spatial
distribution of sand fly species is a first-step for understanding the
ecology of ACL. Environmental suitability for ACL and L. (N.)
whitmani for Brazil was evaluated through ENFA. For data related to
the occurrence of the disease, municipal localities with records of
endemic areas for ACL (N = 1882) were used. For occurrence of
phlebotomines the municipalities with confirmed record of vector (N
= 633) were considered in L. (N.) whitmani Model. The models were
developed using 14 Environmental Variables (EV) (12 bioclimatic,
altitude and NDVI).Cross-validation of the models' quality resulted
in a Boyce index of 0.89 ± 0.10 and 0.84 ± 0.16, for the ACL Model
and the L. (N.) whitmani Model respectively, indicating a satisfactory
predictive capacity of both models. L. (N.) whitmani was identified by
the model as a species that occurs in a particular condition in the study
area, in the context of the average condition of all EVs, and suggests
that the species is moderately tolerant of a range of environmental
conditions (M = 0.87, T = 0.62). Many evidences indicate that this
sand fly has capacity to survive environmental changes and to adapt
to new ecological niches, tolerating effects of climatic alterations.
Analysis by EVs showed that L. (N.) whitmani "prefers" (more
suitable habitat) and is more frequent in regions with relative less
rainfall and warm temperatures. These characteristics are in
accordance with previous analysis discussing the distribution of this
sand fly vector in Brazilian biomes, occurring in high frequency in
Cerrado and Atlantic Rainforest, with greater amplitude in these
Biomes. The ACL Model indicate that more suitable areas for ACL
moderately differs from the average conditions across the entire
distribution area, suggesting that ACL is not too selective of its
environment, and is tolerant of a range of environmental conditions
(M=0.53; T=0.77). The ACT Model identifies more suitable areas
related to intermediate density vegetation (in NDVI EV) and zones
with annual precipitation around 1500 mm.
Financial support: LTL/IOC; Fiocruz/Faperj; Capes.
N° 26 (1) 2015
A FIELD STUDY TO EVALUATE THE PERFORMANCE
OF TWO MODIFIED CDC LIGHT TRAPS FOR
COLLECTING SANDFLIES COMPARED WITH THE
TRADITIONAL CDC TRAP LIGHT
POPULATION COMPOSITION AND DYNAMICS OF
L U T Z O M Y I A L O N G I PA L P I S A N D O T H E R
PHLEBOTOMINAE SPECIES IN CLORINDA,
NORTHERN ARGENTINA
Gould, Ignacio Tomás1, Fernández María Soledad2,3, Pérez Adriana
Alicia4, Salomón Oscar Daniel2,3
Andrea Gómez Bravo1, Ignacio Gould2,4, Marcelo Abril1, Oscar Daniel
Salomón3,4
Redila 1Centro Nacional de Diagnóstico e Investigaciones en Endemoepidemias;
Instituto Nacional de Medicina Tropical; 3Consejo Nacional de Investigaciones
Científicas y Técnicas; 4Departamneto de Ecología, Genética y Evolución, FCEN, UBA
.
Fundación Mundo Sano, Buenos Aires, Argentina; 2. Centro Nacional de Diagnóstico
e Investigaciones Endemo-epidémicas, Buenos Aires, Argentina; 3. Instituto Nacional
de Medicina Tropical, Puerto Iguazú, Misiones, Argentina; 4. Consejo Nacional de
Investigaciones Científicas y Técnicas, CONICET Argentina. Red de Investigación de
Leishmaniasis en Argentina, REDILA
2
odanielsalomon@gmail.com
81
1
agomez@mundosano.org
Keywords: Sanflies, Light Traps, Lutzomyia longipalpis, Nyssomyia
whitmani
Keywords: Lutzomyia longipalpis, Visceral leishmaniasis,
Phlebotominae.
In recent years numerous surveys have been conducted in the
Argentina to study the distribution and dispersal of sandflies due to an
increment in the number of cases of visceral and tegumentary
leishmaniasis. The aim of this work was to compare two local
developed types of light traps for sandflies with the traditional CDC
trap light (CDC). Both news traps are a modified CDC trap light with a
ring of leds, one of white light (WL) and the other of black light (BL).
Traps were compared according to the occurrence and the number of
individuals of sandflies, Lutzomyia longipalpis, the main Leishmania
infantum vector and Nyssomyia whitmani, a L. braziliensis vector. In
order to test the traps 21 sample points were choosing in forest, rural
and urban scenarios in the city of Puerto Iguazú and around it. A Latin
Square design was used for the evaluation of the type of trap. At each
site, the three types of traps were randomly assigned during three
consecutive nights. The variation in the occurrence and numbers of
individuals by species, according to the type of trap, the sex and their
interaction was evaluated using Generalized Linear Mixed Models.
For Lu. longipalpis, 300 individuals captured in 9 sites (urban and
rural environmental) and for Ny. whitmani 1443 individuals captured
in 11 sites (urban, rural and forest environmental) were included in the
analysis. For both species, no differences were found for the
occurrence between the three types of trap for both sex (p>0.05 in all
cases). For Lu. longipalpis, for both sexes, the number of captured
individuals didn´t differ between BL and CDC trap, neither between
CDC and WL trap, but was higher for BL compared to WL trap
(p=0.022). For Ny. whitmani we found no differences between types of
trap for females (p>0.05), but for males we found that the traditional
CDC and the WL traps captured more individuals that the BL traps
(p<0.015 ). These results suggest that the new types of trap could be
used as an alternative to the traditional CDC trap to detect the
occurrence of these species in surveillance studies. In relation to the
estimation of abundance by species and sex, these results shows the
importance of using the same type of trap or combination of traps to
compare species abundances, to avoid biases due to the methodology.
Regarding the design, while a small number of sites were sampling, it
covered a wide range of environments.
In Argentina, the first record of Lu. longipalpis in an urban scenario
was reported in Clorinda city (25°17′29″S 57°43′06″W) during 2004,
locality on the border with Paraguay close to Asunción city that
already has visceral leishmaniasis transmission. The local health
authorities of Clorinda reported cases of canine visceral
leishmaniasis afterward the first vector record. During 2010 a new
entomological survey showed the persistence of Lu. longipalpis in
critical sites of Clorinda. Aimed at describing the spatial and temporal
distribution of Phlebotominae abundance in Clorinda, 20 sites were
sampled on two consecutive nights, from January to December 2012,
using CDC-like light minitraps. The selected sites were peridomestic
habitats in the urban area, based on the Network for Leishmaniasis in
Argentina (REDILA) criteria on 'worst scenario', and
entomoepidemiological background from previous works. The
Phlebotominae collected were separated locally and identified at a
REDILA laboratory. Minimum and maximum temperature values
during sampling days were recorded through a local meteorological
station (CLORICOOP). From January to December 2012, the species
captured were: Lu. longipalpis, Migonemyia migonei, Nyssomyia
whitmani, and Bruptomyia sp. The most abundant species was Lu.
longipalpis 98.73% (4740/4801), followed by Bruptomyia sp 0.62%
(30/4801), Mg. migonei 0.42% (20/4801); and Ny. whitmani 0.23%
(11/4801). Lu. longipalpis was present in all the sampling sites during
the whole year. The month showing the greatest number of
individuals was March (beginning of autumn) with 703
individuals/month, this month being the one when Lu. longipalpis
was most abundant. July (winter) was the month with lower
abundance with a total of 12 individuals, represented by Lu.
longipalpis. During 2012 the average temperatures in Clorinda
ranged between 29.70°C maximum (summer) and 19.39°C minimum
(winter), and the average relative humidity was 79.27%. Lu.
longipalpis turned out to be the species with the highest prevalence in
all the urban area, but with spatial and temporal heterogeneous
abundance due to micro-habitat characteristics and seasonal and daily
climate variations, respectively. Identification of the sites and
moments with the greatest abundance of Lu. longipalpis will result in
a contribution to guide the control measures of this species.
N° 26 (1) 2015
DESCRIPTION OF POPULATION DYNAMICS AND
HOURLY ACTIVITY OF LUTZOMYIA LONGIPALPIS IN
THE CITY OF POSADAS, ARGENTINA
COMPARISON OF CDC LIGHT AND GRAVID TRAPS
FOR THE COLLECTION OF SANDFLY FAUNA IN
BHUBANESWAR, ORISSA (INDIA)
Soraya Alejandra Acardi1, Iris Ingrid Steinhorst2, Maria Soledad
Santini3, Adolfo Enrique Sandoval2, Oscar Daniel Salomon4
Karina Mondragon-Shem1, 2, Melissa Bell1, Mrutyunjay Suar3, Parimita
Routray1, Laura Posada4, Mary Cameron 1
1
1
London School of Hygiene and Tropical Medicine, England, UK | 2 Liverpool School of
Tropical Medicine, England, UK | 3 Kalinga Institute of Industrial Technology â€“KIIT,
Bhubaneswar, Orissa, India | 4 Programa de Estudio y Control de Enfermedades
Tropicales PECET, Universidad de Antioquia, Colombia.
Laboratorio de Biología Molecular Aplicada (LaBiMAp - UNaM). Misiones,
Argentina. 2 Instituto Municipal de Vigilancia y Control de Vectores, Secretaria de
Calidad de Vida, Municipalidad de Posadas. 3 Centro Nacional de Diagnostico e
Investigación en Endemo-Epidemias (CeNDIE) Buenos Aires, Argentina. 4 Instituto
Nacional de Medicina Tropical (INMeT) Puerto Iguazú, Misiones, Argentina.
82
kmshem@liverpool.ac.uk
sorayacardi1@gmail.com
Keywords: Lutzomyia longipalpis, population dynamics, hourly
activity
Lutzomyia longipalpis is the vector of Leishmania infantum in the city
of Posadas, the first urban visceral leishmaniasis focus in Argentina.
To describe its population seasonal dynamics and night activity
phlebotomine sandflies were captured with CDC mini light traps
placed overnight at two sites, two consecutive nights monthly from
February 2009 to March 2010. Then the correlation between average
monthly abundance and temperature, relative humidity (maximum,
minimum and average for both) and rainfall for each month, 30 and 60
days before sampling (DBS) was analyzed by Spearman correlations
(p value <0.05). In three houses with ≤ 30 sandflies a multi-sampling
adapter for CDC mini light traps was placed two consecutive nights at
each site from February to April of 2011, and it was programmed to
change every two hours from 17:00 to 07:00hs. The association of
trap/night average abundance with average temperature (T) and
relative humidity (RH) of each period were studied by Spearman
correlations. Paired t tests were performed to assess differences
between abundance before and after 23:00-1:00hs period (corrected p
value <0.008). Species were identified according to Galati (2003). The
longitudinal study captured 698 Lu. longipalpis (male: female ratio
5.49:1), a bimodal pattern was observed with two peaks of activity,
one in March (end of summer) and another in December (end of
spring). Sandfly abundance showed a positive correlation with
maximum, minimum and average temperature for the month of
sampling and 30 DBS and with maximum temperature 60 DBS. In
hourly activity study 250 Lu. longipalpis were captured (male: female
ratio 5.75:1), the abundance had two peaks of activity at 19:0021:00hs and 1:00- 3:00hs. The least activity was from 23:00 -1:00hs
for all months. Males are present during all periods; females registered
a peak at 19:00-21:00hs period and arrived until 03.00; the 75.8% of
females and 51.5% males was captured during the 17:00-23:00hs
period; and 16.1% of females and 37.08% of males during 1:00-7:00hs
period (no significant difference). There was no correlation of T and H
with total abundance. When only considered females it was positively
correlated with T and negatively with RH in February and March; the
higher abundance was between 32.0°-32.9°C and 50.9%-65.9% RH.
These results showed a trend in the population seasonality and hourly
activity that provides a baseline for the design of protocols on
prevention- control strategies research.
Keywords: CDC trap, India, Phlebotomus argentipes
Increasing distribution of sandfly-borne diseases makes
entomological surveillance fundamental to determine changes in
vector species and define transmission risks. The CDC light trap (LT)
is one of the most commonly used sampling methods for sandflies. On
the other hand, CDC gravid traps (GT), normally used for mosquito
sampling, have recently been found to collect phlebotomine sandflies
in Tanzania and India. In this work we set out to compare the efficacy
of the LT vs GT for the collection of sandflies in Bhubaneswar, Orissa.
Sampling was carried out using a Latin Square design in the Salia Sahi
slum in Bhubaneswar, placing five traps of each type indoors in 10
households during 30 nights (=300 trap-nights). In addition, we tested
the efficacy of both traps in a buffalo farm for 24 nights. Sandflies
were identified to species, making note of male genitalia rotation and
female physiological status (unfed, fed, and gravid). Negative
binomial regression was used to analyse the over dispersed catch data
and incidence rate ratios were calculated to measure treatment effects.
We report the first evaluation of GT for the collection of sandflies, and
the first evaluation of this trap indoors. In households, LT was five
times more likely to collect sandflies than the GT, nearly five times
more likely to collect females (including gravid females), and three
times more likely to collect males. Both traps performed similarly in
the buffalo farm. The sandfly species identified include Phlebotomus
argentipes, Sergentomyia antennata, S. babu and S.squamipleuris.
Our results show the LT outperformed the GT in all aspects, and
remains one of the best options for entomologicalsurveillance of
sandfly populations. We report the intradomiciliary presence of
Phlebotomus argentipes in Orissa state. This sandfly vectors
Leishmania donovani, which causes anthroponotic visceral
leishmaniasis in India. Although no VL cases are reported in Orissa,
frequent human migration is a potential threat to the introduction of
the parasite into the state from the well-known endemic areas of
Bihar, Jharkhand or West Bengal.
N° 26 (1) 2015
83
ID 47-P Taxonomy & Phylogeny
TAXONOMY AND DIAGNOSTIC CHARACTERS OF
THE PHLEBOTOMUS ARGENTIPES SPECIES
COMPLEX
DISCRIMINANT ANALYSIS AS A POSSIBLE TOOL TO
I D E N T I F I C AT I O N S A N D F L I E S ( D I P T E R A :
PSYCHODIDAE) SPECIES THROUGH
MORPHOMETRIC DATA
Kandan Ilango
Zoological Survey of India Zoological Survey of India Ministry of Environment &
Forests Government of India Southern Regional Centre Chennai, INDIA
kilangozsi@gmail.com
Keywords: Phlebotomus argentipes, taxonomy, species complex,
visceral leishmaniasis, India
Visceral Leishmanisis (VL) (Kala-Azar) transmitted by the bite of
sand fly, Phlebotomus argentipes sensu lato is endemic in parts of
India, Nepal and Bangladesh accounting for 90% of the global disease
burden. Ph. argentipes s. l. occurs widely in southern Asia exceeding
the limits of disease endemic regions but its population's exhibit
differences in biology and behaviour. The taxonomy of Ph.argentipes
s.l. based on descriptive morphology and principal component
analysis reveals unequivocally 3 sibling species viz. Ph. argentipes
sensu stricto, Ph. annandalei and Ph. glaucus from India. It remains
however unknown which of the three sibling species is the Indian VL
vector. A morphologically distinct population of Ph. argentipes from
Sri Lanka that differs from the members of Indian species complex of
Ph. argentipes exists in areas of active cutaneous leishmanisis
transmission. Molecular identification of the Sri Lankan Ph.
argentipes using sequences of COI, cyt b, ITS2 and 28s rDNA
confirmed the existence two sibling species. Vector control in Indian
VL elimination program needs urgently the identification and
characterization of members of the Ph. argentipes species complex.
Rodrigo E. Godoy1; Eunice A. B. Galati2, Nataly A. de Souza1, Thiago V.
dos Santos3; Lindemberg Caranha de Sousa4; Adriana Zwetsch1;
Elizabeth F. Rangel1
1
Instituto Oswaldo Cruz - Fundação Oswaldo Cruz; 2Faculdade de Saúde Pública –
Universidade de São Paulo; 3 Instituto Evandro Chagas; 4Secretaria de Saúde do
Estado do Ceará
elizabethrangel@fiocruz.br
Keywords: Sand flies, Taxonomy, linear morphometrics
The sand flies taxonomy based in the linear morphometry and
morphology of the adult insect still presents some problems as for the
determination of species, especially regarding the females
specimens, where more than 150 from a total of 500 species needs the
presence of a male specimen to fully determinate the right species.
Besides that the presence of species complex it is a further
complicating factor for the identification in some genera. Thus our
work aimed to use discriminant analysis through morphometric data
to evaluate the capacity of this analysis to properly determinate the
sand flies species. We evaluate five sand flies species (40 specimens
for each species), that are proven Leishmaniasis vectors, where:
Nyssomyia intermedia, Ny. whitmani, Bichromomyia flaviscutellata,
Migonemyia migonei, Ny. neivai, Brumptomyia troglodytes (used as
control group). To cluster the species through the morphometric data
we performed 2 different analyzes, one with the characters present
both in male and female (shared characters) more the specifics from
male (120 specimens) using 48 characters, and the other with the
shared characters more the specifics from female (120 specimens)
using 43 characters. Two statistical tests, canonical variables and
discriminant function were used to obtain the best clustering among
species groups. In this analysis, also the values of F and Wilks'
Lambda, were evaluated. The characteristics that most contributed to
the species differentiation in the 110 males specimens were the
structures of the terminalia. All the species presented 100% of correct
classification even for the cross-validation, nevertheless between Ny.
intermedia and Ny. neivai, after the cross-validation, the correct
classification was respectively: 85% (17/20 specimens) and 90%
(18/20). The characters that most contributed to the differentiation of
the species in the 114 females specimens were the characters shared
between males and females. All the species presented 100% of correct
classification even for the cross-validation. Nevertheless between Ny.
intermedia and Ny. neivai, after the cross-validation, the correct
classification was respectively: 85% (17/20 specimens) and 62.5%
(10/16). The discriminant analysis using morphometric data can be
used as an additional tool for the definition of cryptic species, helping
to define species.
N° 26 (1) 2015
84
FIRST REPORT OF ABNORMAL NUMBERS OF
SPERMATHECAE IN A FEMALE PHLEBOTOMUS
(LARROUSSIUS) LONGICUSPIS (DIPTERA:
PSYCHODIDAE) IN TUNISIA
FIRST RECORD OF PSATHYROMYIA
P R A D O B A R R I E N TO S I ( L E P O N T, M AT I A S ,
MARTINEZ & DUJARDIN, 2004) (DIPTERA:
PSYCHODIDAE: PHLEBOTOMINAE) IN BRAZIL
Jomaa Chemkhi1, Souheila Guerbouj1, Ikram Guizani1, Afif Ben Salah2
Douglas de Almeida Rocha, Jônatas Barbosa Cavalcante Ferreira,
Rodrigo Gurgel-Gonçalves & Andrey José de Andrade
1
Laboratoire d'épidémiologie Moléculaire et Pathologie Expérimentale Appliquée aux
Maladies infectieuses, Institut Pasteur de Tunis, Université Tunis El Manar, Tunisia;
2
Laboratoire d'épidémiologie Médicale, Institut Pasteur de Tunis, Université Tunis El
Manar, Tunisia.
Laboratório de Parasitologia Médica e Biologia de Vetores, Área de Patologia,
Faculdade de Medicina, Universidade de Brasília, Campus Universitário Darcy
Ribeiro, Asa Norte, 70910-900, Distrito Federal, Brazil.
jomaa.chemkhi@pasteur.rns.tn
dougalmeidarocha@gmail.com
Keywords: Phlebotomine sand fly, spermathecae, abnormality
Keywords: Psathyromyia runoides, P. prabobarrientosi,
geographical distribution, taxonomy
Phlebotomine sand flies (Diptera: Psychodidae) constitute a large
group of flies including about 800 species, many of which have been
described as vectors of leishmaniasis, a disease caused by Leishmania
(Kinetoplastida: Trypanosomatidae) parasites. Species morphological
identification is based for males, on the genitalia properties, while it is
based mainly on specific characteristics of the spermathecae and
ducts, for female specimens. In Tunisia, a total of 17 species belonging
to the genus Phlebotomus and Sergentomyia were described. In this
work we report an abnormality found in the spermathecae of a female
specimen of the species Phlebotomus (Larroussius) longicuspis. Sand
flies were collected in the locality of Sidi Saad, in the governorate of
Kairouan, central Tunisia, during the period from June to November
2011. CDC light traps were placed overnight, in different biotopes
(inside habitations, outdoors and within animal shelters). Specimens
were collected and males were washed, mounted and directly
identified under a microscope. Females were either dissected and
identified or stored in liquid nitrogen for later identification.
Dissection of female sand flies was carried out in sterile saline
solution. The last two abdominal segments containing spermathecae
and ducts were mounted for morphological identification based on the
keys of Lewis (1982) and Léger et al. (1983). During the survey of
sand fly fauna in the above mentioned region, 163 flies were
morphologically described and the species identified, in the locality of
Sidi Saad, within the animal shelters biotope. Among the studied sand
fly population we found an abnormal spermathecae in the case of a
female specimen identified as belonging to the species Phlebotomus
(Larroussius) longicuspis. This specimen presented with three
spermathecae. Indeed, an additional spermathecae located in a short
bifurcation on the upper extremity of one duct was found. This
abnormality is considered rare according to the described anatomy of
phlebotomine sand flies. The abnormality found in the female
spermathecae of the species Phlebotomus (Larroussius) longicuspis is
for the first time described in Tunisia. Morphological abnormalities
are important to report in order to avoid erroneous sand fly
identification or description of new species. This study received
financial support from the Tropical Medicine Research Centers (P50)
program (Contract No. RFA-AI-06-006), USA.
Psathyromyia pradobarrientosi (Le Pont, Matias, Martinez &
Dujardin, 2004) was described based on males from Bolivia and
nowadays this is the single record of the species. It is morphologically
similar to P. runoides (Fairchild & Hertig, 1953), which was
described in Panama. Males of these phlebotomine sand fly species
can be distinguished based on the ejaculatory duct length in relation to
the pump and its apex. The female of P. pradobarrientosi is
undescribed. Recently, two males of sand flies identified as
Psathyromyia sp. were collected in forested areas of Brasília, Federal
District, Brazil. To access the taxonomic identity of these specimens
they were compared with the male of P. runoides paratype deposited
in the Coleção do Museu de Zoologia da Universidade de São Paulo
and with the original descriptions of P. pradobarrientosi and P.
runoides. The collected specimens and the types of P. runoides were
measured, drawn and photographed. Males of Psathyromyia sp.
presented the following characteristics: 1) the ejaculatory duct was 3
times longer than the ejaculatory apodeme (piston + sperm sac); 2) the
apex of the ejaculatory duct was inflated at end; 3) the ventral margin
of the basiphallus was 2 times smaller than that of P. runoides; 4) the
bristles on the dorsal margin of the paramere were distributed at least
on the apical half. Based on these characteristics the males of
Psathyromyia sp. were identified as P. pradobarrientosi and this
species is registered for the first time in Brazil. Due to morphological
similarity between P. pradobarrientosi and P. runoides, the
geographical distributions of both species should be revised. We
hypothesize that Psathyromyia runoides presents a trans-Andean
distribution while P. pradobarrientosi is a cis-Andean species in
South America.
N° 26 (1) 2015
RECORDS OF FEMALES PSATHYROMYIA CUZQUENA
MARTINS, LLANOS & SILVA, 1975 (DIPTERA,
PSYCHODIDAE, PHLEBOTOMINAE) IN NORTHERN
BRAZIL
ASSESSMENT OF THE TAXONOMIC STATUS OF
PHLEBOTOMUS PIFANOI ORTIZ, 1972 (DIPTERA,
PSYCHODIDAE, PHLEBOTOMINAE)
Priscila Bassan Sábio, Andrey José de Andrade and Eunice Aparecida
Bianchi Galati
85
Priscila Bassan Sábio, Andrey José de Andrade and Eunice Aparecida
Bianchi Galati
Faculdade de Saúde Pública, Universidade de São Paulo
priscilabassan@usp.br
Keywords: Phlebotominae, Sand fly, Psathyromyia cuzquena,
Keywords: Phlebotominae, sand flies, Shannoni Complex,
Phlebotomus pifanoi
Misidentification
Psathyromyia cuzquena (Martins, Llanos & Silva, 1975) was
described on the basis of two males collected in Pilcopata, Cuzco
Department, Peru. It belongs to Shannoni Series and genus and
subgenus Psathyromyia. This species was considered by the authors
morphologically close to Pa. shannoni (Dyar, 1929) and Pa.
souzacastroi (Damasceno & Causey, 1944) and is also distributed in
Amazonian Brazil, Colombia and Venezuela. In the identification
keys of American sand fly species it has only been possible to identify
the male of the Pa. cuzquena, since the female has never been
described. However, some fauna surveys have reported the female of
Pa. cuzquena in northern Brazil, along with other species of the
Shannoni Series. These studies were conducted in the municipalities
of Borba, Maués, Nhamundá and Manaus, Amazonas state, and
Caroebe, Roraima state. Literature searches were undertaken in the
Medline, Scopus and SciELO online databases for records of the
description of the female of Pa. cuzquena, and some depository
collections of species sand fly were consulted, but nothing was found.
Sand fly are a group of insects of great importance to public health due
to the haematophagic habit of their females that bite human beings and
other animals and can transmit infective agents, such as trypanosomes,
especially the various species of the genus Leishmania.
Misidentifications of species may lead to false geographical records,
thus confounding the understanding of the role of the vectors involved
in the transmission of diseases.
Psathyromyia shannoni (Dyar, 1929) had Phlebotomus pifanoi Ortiz,
1972, known only by its male holotype, considered as its junior
synonym – though without any justification – by Martins, Williams
and Falcão (1978). Psathyromyia cuzquena (Martins, Llanos & Silva,
1975) was described from a single male, whose female is unknown.
The taxonomic status of Ph. pifanoi is here reviewed on the basis of its
morphological characters reported in the literature and compared
with those of Pa. shannoni and close species deposited in
entomological collections and the literature. Psathyromyia shannoni
and Ph. pifanoi are morphologically distinct species and therefore not
synonymous. In the gonostyle of Ph. pifanoi, the upper external spine
is implanted closer to the apical spine than to the external lower spine,
while in Pa. shannoni the upper external spine is implanted
equidistant from the apical spine and the lower external spine. In the
paramere of Ph. pifanoi the bristles on the dorsal edge extend from its
apex to the level of the most apical bristle of the ventral margin and in
Pa. shannoni the bristles on the dorsal edge extend from its apex to the
level of the most basal bristles of the ventral margin. On the other
hand, Pa. cuzquena is morphologically identical to Ph. pifanoi. Thus,
Psathyromyia pifanoi stat. rev.; comb. nov. is a valid species and in
accordance with the criterion of priority it is the senior synonym of
Pa. cuzquena sin. nov.
N° 26 (1) 2015
86
UNCHARTED PHLEBOTOMINE SAND FLIES IN
GABON
MORPHOMETRIC STUDY ON POPULATIONS OF
PSYCHODOPYGUS COMPLEXA AND PS. WELLCOMEI
( D I P T E R A : P S Y C H O D I D A E ) , V E C TO R S O F
CUTANEOUS LEISHMANIASIS IN BRAZIL
Nil Rahola
IRD MIVEGEC unit (France) & CIRMF UEM unit (Gabon). nil.rahola@ird.fr
Keywords: new species taxonomy update
Patrick Feitosa Botelho¹, Fábio Márcio Medeiros da Silva¹, Rodrigo
Espíndola Godoy², Thiago Vasconcelos dos Santos¹
1
Instituto Evandro Chagas Svs Ms, Ananindeua, Pa, Brazil
thiagovasconcelos@iec.pa.gov.br
Despite the former mention of a native case of visceral leishmaniasis,
Gabon does not currently constitute an endemic country for this
disease. As a result, the sandfly fauna in this country remains poorly
documented and prospected. An exhaustive review of literature
reports only four species in Gabon. From 2012 to the end of 2013 we
managed to collect more than 5000 sand flies through a large mosquito
survey that consisted in 1850 CDC miniature light traps installed in
two different forest sites and 560 CDC miniature light traps installed
in two different caves. Over 1500 specimens were found in forest sites
and 3500 in caves. In addition, some collections were performed using
other CDC miniature light traps in anthropic environments such as
small savannahs villages, forest villages and towns. Those collections
allowed us to bring the current checklist for Phlebotomine sand flies of
Gabon from four to 25 species, and add at least three new species for
Science (currently being described). The forest was the highest spot
for species diversity and allowed us to discover and describe a new
species of Phlebotomus with a quite singular morphology:
Phlebotomus (Legeromyia) multihamatus and also the unknown male
of Spelaeomyia moucheti. This discovery will lead us to perform a
molecular analysis of the whole genus Spelaeomyia. On the other
hand, in caves, sand flies were very abundant but of poor diversity.
With those new collections we will be able to bring new
morphological data and make some re-descriptions of some
specimens such as the female of Spelaeomyia moucheti or
Sergentomyia lumsdeni, but also to consider the possibility of the
creation of new subgenus for the genus Sergentomyia. This update of
the Phlebotomine sand flies of Gabon must encourage prospecting in
this country. Even if only one case of leishmaniasis has been reported
in Gabon, the role of sand flies in the transmission of such parasites or
other infectious agents (such as viruses or bacteria), to wildlife,
domestic animals or to humans, as well as their trophic preferences,
should be evaluated.
Keywords: Morphometrics, Psychodopygus, cutaneous
leishmaniasis
The correct identification of sand fly species is an important
component for strategies of surveillance and leishmaniasis control.
Despite medical relevance, the females of Psychodopygus from
chagasi series are so far morphologically indistinguishable. Among
these species, Psychodopygus wellcomei and Ps. complexa are
respectively incriminated and suspected in the transmission of
cutaneous leishmaniasis at Amazonian Brazil. The aim of this work is
to analyze different populations of Lu. (Ps.) complexa and Lu. (Ps.)
wellcomei by classical and geometric morphometry and thus attempt
to discriminate them. It was selected 104 individuals from the
“Coleção de Flebotomíneos do IEC” (ColFleb IEC) mounted on glass
slides with berlese liquid and distributed as follow: Ps. complexa
from Vitória do Xingu- Pará State (PA) (20 Males+ 20 Females), Ps.
wellcomei from Parauapebas- PA (25M+ 25F), Ps. wellcomei from
Baturité- Ceará State (CE) (14M+ 14F). Their correct identification
was ensured for the absence of simpatry on the collects. An external
group was composed by individuals of Brumptomyia sp. from
Ananindeua- PA (3F). The measurements were made with a
computer- assisted system (Axiovision- Zeiss). For classical
morphometry, 15 characters were selected for females and 19 for
males. For geometrical morphometry, 5 anatomic marks were used
from the right wing of each specimen. The analysis was made with
canonical variables and discriminant function. The classical
morphometry allowed distinguish the species with 89,8% of correct
classification for males and 69,4% for females at cross validation
(88% for Ps. complexa PA, 40% for Ps. wellcomei PA and 66.7% for
Ps. wellcomei CE). The geometric morphometry showed short
distance (3,0044) between the Psychodopygus spp. in comparison of
the external group (10,0677). The female specimens were
distinguished with 93, 3% of correct classification in cross validation.
In conclusion, although a high probability of correct identification
was achieved with geometric morphometry, not all females can be
distinguished unambiguously with both methods.
N° 26 (1) 2015
87
TAXONOMIC REVIEW OF PSYCHODOPYGUS
MANGABEIRA, 1941 (DIPTERA, PSYCHODIDAE,
PHLEBOTOMINAE)
TAXONOMIC REVISION OF THE SUBGENUS
PSATHYROMYIA BARRETTO, 1962 (DIPTERA:
PSYCHODIDAE: PHLEBOTOMINAE)
Veracilda Ribeiro Alves1, Claudio Ruy V. da Fonseca2, Paloma Helena F.
Shimabukuro1
Priscila Bassan Sábio, Andrey José de Andrade e Eunice Aparecida
Bianchi Galati
1
Centro de Referência Nacional e Internacional de Flebotomíneos/Centro de Pesquisas
René Rachou/FIOCRUZ, Belo Horizonte-MG, Brazil. 2 Coordenação de
Biodiversidade/Instituto Nacional de Pesquisas da Amazônia/INPA, Manaus-AM,
Brazil.
verabioufmt@gmail.com
Keywords: Phlebotominae, Psathyromyia, Sandfly, Taxonomy
Keywords: Morphological characters, Psychodopygus, sand flies;
Neotropical region
Phlebotominae sandfly (Diptera, Psychodidae, Phlebotominae) are a
group of public health concern due to their haematophagic habit of
females that bite man and other animals, causing them annoying and
they can also transmit infection agents such as trypanosomes,
especially for the various species of the genus Leishmania, bacteria
and viruses. Identification of American sandfly is traditionally based
on morphology. However some species are morphologically very
similar, leading to errors in identification, as in the species included in
the subgenus Psathyromyia. Because of this similarity, especially
among females, this subgenus had some of their species considered as
junior synonyms of Psathyromyia shannoni. Some of these were
revalidated in a previous study. Thus, this project aims to review the
taxonomic status of the species of the subgenus Psathyromyia by
means of morphological, morphometric and molecular analyzes. The
use of molecular studies, together with morphological can help in the
correct diagnosis of species of this subgenus. With this, we hope to
contribute to a better understanding of the role epidemiological and
vector of these species in Brazil.
The species of the genus Psychodopygus Mangabeira, 1941 occur in
woods and forests. In human bait collections carried out in the
Amazon region at the forest floor level, most species of sand flies
collected belong to this genus. The females of some species are
difficult to identify due to the existing interspecific morphological
similarity, therefore correct identification is only possible when they
are collected together or in copulation. The species are predominantly
neotropical occurring from southern Mexico to southern Brazil with
over 80% of the species recorded in the Amazon region. The species of
this genus are of medical interest because they are vectors of
Leishmania and are highly anthropophilic. The revision of the species
was based on the examination of type specimens deposited in the
following biological collections: Faculdade de Saúde
Pública(FSP)/SP, Instituto Evandro Chagas (IEC)/PA, Museu de
Zoologia da USP (MUSP)/SP, Instituto Oswaldo Cruz (IOC)/RJ,
Instituto Nacional de Pesquisas da Amazônia (INPA)/AM, Centro de
Pesquisas René Rachou CPqRR)/MG, Walter Reed Biosystematics
Unit (WRBU)/Smithsonian Institute/USA and Institut de Recherché
pour le Développement (IRD)/France. At the beginning of this study
the genus Psychodopygus comprised 39 valid species and of these 24
type-species were examined. The types of nine species could not be
examined due to difficulties in contacting those responsible for the
collections in which they were deposited; six types have been lost: Ps.
fairchildi (Barretto, 1966), Ps. guyanensis (Floch & Abonnenc,
1941)¸ Ps. hirsutus (Mangabeira, 1942), Ps. complexus (Mangabeira,
1941), Ps. geniculatus (Mangabeira, 1941) and Ps. parimaensis (Ortiz
e Álvares, 1972). The informally described species of Três Esquinas
morphotypes were confirmed as being Ps. guyanensis.
Psychodopygus douradoi (Freitas & Fé, 1998) is proposed as junior
synonynym of Ps. fairtigi (Martins, 1970), and Ps. parimaensis as a
junior synonynym of Ps. davisi (Root, 1934). Thus, the number of
valid species after this study is 36. This revision revealed the status of
the type species deposited in different institutions and the importance
of examining them for comparison with other similar species and
descriptions of new species.
N° 26 (1) 2015
DNA BARCODING FOR THE IDENTIFICATION OF
SAND FLY SPECIES (DIPTERA, PSYCHODIDAE,
PHLEBOTOMINAE) IN PORTUGAL
NEW RECORDS OF PHLEBOTOMINAE (DIPTERA:
PSYCHODIDAE) FROM THE ATLANTIC FOREST
REGION OF ARGENTINA
C. Maia1,2,3, R. Parreira4,5, J. Cristóvão 1,2, M.O. Afonso1,5, L. Campino1,6
Casertano,S.A.1; Andrade Filho, J.D2; Salomon, O.D1,3
1-
Unidade de Parasitologia Médica, Instituto de Higiene e Medicina Tropical (IHMT),
Universidade Nova de Lisboa (UNL), Lisboa, Portugal 2-Centro de Malária e outras
Doenças Tropicais 3-Faculdade de Medicina Veterinária, Universidade Lusófona de
Humanidades e Tecnologias, Lisboa, Portugal 4-Unidade de Microbiologia Médica,
IHMT, UNL 5-Unidade de Parasitologia e Microbiologia Médicas 6-Departamento de
Ciências Biomédicas e Medicina, Universidade do Algarve, Faro, Portugal
carlamaia@ihmt.unl.pt
Keywords: Phlebotomine sand flies, Barcoding, Cytochrome oxidase
gene subunit I, Portugal
Phlebotomine sand flies (Diptera, Psychodidae) transmit pathogens
that affect humans and animals worldwide. As sand fly genera and
species vary in geographical distribution, ecology, and pathogen
transmission, surveillance of their presence and distribution are
important. Five sand fly species are known to be present in Portugal:
Phlebotomus ariasi, P. papatasi, P. pernicious, P. sergenti and
Sergentomyia minuta. P. perniciosus and P. ariasi are the proven
vectors of Leishmania infantum while L. major DNA was recently
detected in one S. minuta specimen collected in the south of the
country. Although the identification of sand flies is currently based on
morphological characters, it requires considerable skill and
taxonomic expertise. DNA-based approaches are useful tools for
estimating sand fly diversity and for ensuring an accurate
identification of species. A partial sequence of the mitochondrial
cytochrome oxidase gene subunit I is currently being used to
differentiate species in different animal taxa, including insects. The
aim of this study was to evaluate the utility of the DNA barcoding for
the identification of phlebotomine sand flies in Portugal. P. ariasi, P.
pernicious, P. sergenti and S. minuta specimens collected from the
North, Center and South of the country were morphologically
identified prior to their use for barcoding. Analyses of the sequences
generated indicated that the observed species clusters were consistent
with the morphological identifications. In conclusion, barcoding is a
useful tool for discriminating the sand fly species existent not only in
Portugal but also in the Mediterranean basin, and in the future can
probably be used as an extra and efficacious tool for monitoring the
presence and spread of the different sand fly species responsible for
the transmission of pathogens of medical and veterinary concern.
Funding: This work was partially supported by EDENext (FP7INCO-CT-2010-261504). C. Maia (SFRH/BPD/44082/2008) holds a
fellowship from Fundação para a Ciência e a Tecnologia, Ministério
da Ciência, Tecnologia e Ensino Superior, Portugal.
88
1
Instituto Nacional de Medicina Tropical - INMeT, Puerto Iguazú Argentina,REDILA;
Centro de Pesquisas René Rachou - FIOCRUZ, Belo Horizonte, Minas Gerais, Brazil;
3
CONICET, Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina
2
casertanoameri@yahoo.com.ar
Keywords: new records, Argentina, phlebotomine species, Atlantic
Forest
The Atlantic Forest is one of the most endangered rainforests on earth,
of which remains about 7% of its original extension. Misiones
Province in Argentina and neighboring areas of Brazil and Paraguay
contain the “Green Corridor”, the largest remnant of this forest that,
together with the populated areas, also constitute one of the most
important geopolitical, economical and cultural cross points of South
America. Recent increasing sampling efforts in studying
Phlebotomine sand flies communities in different anthropic and nonanthropic environments of the Atlantic Forest of Argentina have
resulted in new records for the region. Four species have been found
in natural protected areas (Iguazú National Park) and in forest patches
remaining in recently occupied areas, both in Puerto Iguazú
municipality, north of Misiones Province: Micropygomyia capixaba
(Dias, Falcão, Silva & Martins, 1987); Evandromyia correalimai
(Martins, Coutinho & Luz, 1970); Evandromyia edwardsi
(Mangabeira, 1941) and Evandromyia bourrouli (Barreto &
Coutinho, 1941). In rural environments nearby San Pedro city, central
Misiones Province mountain area, Martinsmyia alphabetica
(Fonseca, 1936) was found, more than 60 years after its last record.
The main landscape out protected areas consisted in little farms
containing small patches of rainforest. All these species have been
also found in neighboring Brazilian State of Paraná in the same
semidecidual rainforest, and M. alphabetica have been also reported
in urban areas of Brazilian neighboring city of Foz do Iguaçú. These
results, together with recent findings in Chaco Region, increase
Argentina's Phlebotomine species number up to 36.
N° 26 (1) 2015
89
A METHOD FOR EXTRACTION OF GENETIC
MATERIAL WHICH PERMITS THE INTEGRATION
OF CLASSICAL TAXONOMY WITH IDENTIFICATION
BASED ON THE DNA BARCODE
CONTRIBUTION TO THE KNOWLEDGE OF THE
FAUNA OF SAND FLIES (DIPTERA: PSYCHODIDAE:
PHLEBOTOMINAE) OF RIO DOCE STATE PARK,
MINAS GERAIS, BRAZIL
Alveiro Pérez-Doria, Luís Romero-Ricardo, Natalia Lastre, Eduar Elías
Bejarano
Cristian Ferreira Souza1; Reginaldo Peçanha Brazil2; José Dilermando
Andrade Filho3; Paula Dias Bevilacqua4; Patrícia Flávia Quaresma3;
Mirelle Barbosa Souza5
Grupo de Investigaciones Biomédicas, Universidad de Sucre, Sincelejo, Colombia.
Keywords: DNA Extraction, Sandflies, classical taxonomy, DNA
barcoding
In the study of phlebotomine sand flies molecular biology techniques
have provided valuable evidence that has helped resolve taxonomic
problems both at the infraspecific and supraspecific levels.
Nevertheless, in most of these studies part or even all of the insect
body is used to extract the genetic material which is subsequently used
for genotyping. Thus, although analysis of the sequences generated
may resolve the research question, the voucher specimens required for
classical taxonomy remain incomplete due to the loss of
morphological characters. With this problem in mind, in the present
study the concentration and quality of the DNA obtained with a simple
extraction method based on high salt concentrations and enzymatic
maceration were evaluated, this technique yielding the genetic
material of the insect while allowing the exoskeleton to be recovered
intact for use as a voucher specimen. Phlebotomines were collected in
Colosó, in the Colombian departament of Sucre. The effects of
different concentrations of proteinase K and distinct incubation times
on the final concentration of DNA were determined by measuring in a
spectrophotometer. Integrity of was evaluated by amplifying and
sequencing a 550 bp fragment of mitochondrial DNA. The DNA
concentrations varied between 12 and 236 ng/ l, levels that were
markedly higher than those obtained for the group processed by
mechanical maceration (15 - 42 ng/ l). The barcode sequences of the
cytochrome b gene of Lu. evansi, Lu. panamensis, Lu. gomezi, Lu
longipalpis, Lu. dubitans, Lu. trinidadensis, Lu. micropyga, Lu.
cayennensis cayennensis, Lu. venezuelensis and Lu. atroclavata were
obtained. The exoskeletons were recovered intact and mounted in
Canada balsam. These results provide entomologists with a useful tool
for the morphological and genetic identification of species, permitting
the integration of classical and molecular taxonomy.
1
Programa de Pós-Graduação em Biologia Parasitária - Instituto Oswaldo Cruz Fundação Oswaldo Cruz – Av. Brasil, 4365 – Manguinhos, CEP: 21040-360. Rio de
Janeiro–RJ. Brazil; 2Laboratório de Doenças Parasitarias - Instituto Oswaldo Cruz Fundação Oswaldo Cruz - Manguinhos,RJ. Brazil; 3Centro de Referência Nacional e
Internacional para Flebotomíneos - Centro de Pesquisas RenéRachou/Fiocruz, Av.
Augusto de Lima, 1715 - Barro Preto, CP1743. CEP: 30190-002. Belo Horizonte–MG.
Brazil; 4Departamento de Veterinária - Universidade Federal de Viçosa, Av. Peter
Henry Rolfs, s/n - Campus Universitário. CEP: 36570-000. Viçosa–MG. Brazil;
5
Graduação em Ciências Biológicas - Universidade Federal de Viçosa, Av. Peter
Henry Rolfs, s/n - Campus Universitário. CEP: 36570-000. Viçosa–MG. Brazil.
biominas2004@yahoo.com.br
Keywords: sand flies, vectors, Leishmaniasis
Currently about 500 species of sand flies are known in the Americas,
and only in Brazil have been recorded 267 of these species. This study
aimed to identify the sand fly fauna found at the Parque Estadual do
Rio Doce (PERD), located in the eastern state of Minas Gerais,
Brazil, between the municipalities of Timóteo Marliéria and
Dionísio. The collections were made in Juquita Trail, located in the
PERD area belonging to the municipality of Timóteo-MG, and the
sand flies of the urban area of the municipality is already known.
During collection 12 CDC light traps, installed from September 2012
to February 2014 were used. Traps were exposed for two consecutive
nights, at a height of 150 cm from the floor nights. To date a total of
994 sand flies belonging to 20 species were identified, 693 (69.7%)
male and 301 (30.3%) females, were: Brumptomyia avellari,
Brumptomyia cunhai, Brumptomyia nitzulescui, Evandromyia
cortelezzii, Evandromyia sallesi, Evandromyia termitophila,
Evandromyia tupynambai, Micropygomyia borgmeirai, capixaba
Micropygomyia, Micropygomyia quinquefer, Migonemyia migonei,
Nyssomyia intermedia, Nyssomyia Whitmani, Pintomyia fischeri,
Pintomyia pessoai, Pressatia choti, Psathyromyia lanei,
Psathyromyia pascalei, Psychodopygus carreirai, Psychodopygus
davisi, Sciopemyia sordellii and Trichopygomyia longispina. Among
the species caught Pressatia choti (21.3%) Trichopygomya
longispina (16.1%) and Nyssomyia intermedia (14.2%) were the
most abundant species. It is very important to report the presence of
Nyssomyia whitmani (5.2%), Migonemyia migonei (3.0%), and
Nyssomyia intermedia species already recognized as vector of
Leishmania. Importantly, the PERD is located in an endemic area of
leishmaniasis, with cases already recorded among employees and
park visitors. The identification of these species as possible vectors,
help to better understand the cycle of transmission of leishmaniasis
within the Park boundaries.
N° 26 (1) 2015
90
SAND FLY FAUNA IN KALEYBAR DISTRICTS,
NORTHWESTERN IRAN
MALFORMATIONS OF THE GENITALIA IN MALE
MICROPYGOMYIA QUINQUEFER (DYAR, 1929)
(DIPTERA: PSYCHODIDAE) IN CORRIENTES,
ARGENTINA
Amini Esfanjani MH1, Barazesh A1, Mazloumi Gavagni A1,SaeidiZ 2*
1
Department of Medical Parasitology, Faculty of Medicine, Research Center for
Infectious and Tropical Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
2
Department of Medical Entomology and Vector Control, School of Public Health,
Tehran University of Medical Sciences, Tehran, Iran.
saeidizs@yahoo.com
Mirta Liliana Mierez, María Josefa Felisa Rea, Carlos
Borda, Luis Armando Mosqueda
Edgardo
Centro Nacional de Parasitología y Enfermedades Tropicales (Cenpetrop), Facultad
de Medicina, Universidad Nacional del Nordeste, Santa Fé 1432, 3400 Corrientes,
Argentina
cenpetrop@hotmail.com
Keywords: visceral leishmaniasis, sand fly, Kaleybar, Iran
Introduction: In Iran Leishmania infantum is the main causative agent
of the visceral leishmaniasis (VL). Studying on vector species is
valuable for epidemiological studies and for planning control
mesures. So this study was carried out to diagnose and determine the
sand fly species in eight villages of Kaleybar. Methods: Four VL
endemic villages and four none endemic villages were selected from
Kaleybar district, light traps and in some occasions sticky papers were
used to collect sand flies. They were mounted using lactophenol and
Pouri's media and identified by the use of valid identification keys.
Results: A total of 5000 sand flies were collected and 500 sand flies
were identified. The study showed that subgenus Larroussius of
Phlebotomus, in particular P. perfiliewi, P. tobbi and P. kandelakii,
among endemic villages and P. (Paraphlebotomus) sergenti among
none endemic villages were the most abundant. Discussion: It seems
that P. (Larroussius) perfiliewi was the most abundant species in the
VL endemic areas and P. (Larroussius) tobbi and P. (Larroussius)
kandelakii were in second rank. Other studies in Northwestern
endemic foci of Iran also reported similar results. Such researches
would be crucial in this area because there are some nomadic tribes
which maybe could play a role to transfer the disease to different
areas with various climates.
Keywords: Malformations, male, Micropygomyia quinquefer,
Argentina
A preliminary study indicated a high prevalence of dermal
leishmaniasis in the human population of the Corrientes province.
Based on the reported cases of human infections of cutaneous
leishmaniasis, the rural district of Itatí was selected for the
entomological studies. Nyssomyia neivai, Migonemyia migonei and
Micropygomyia quinquefer were collected in a sylvatic habitat of Itatí
(Borda et al. 2006).The predominance of Mi. quinquefer and its
abundance in this area agree with other studies that indicated that this
species is in the process of adapting to modified environments
especially during the day. Mi. quinquefer belong to a group that was
described with five well-developed spines on the style, of which two
are terminal, among other morphological characteristics.
Morphological characters present in the genitalia of phlebotomine are
essential for correct identification of species. The taxonomic
identification of the Phlebotominae associated with the
epidemiological cycle of leishmaniasis is based on morphological
characters of the adult insect. Changes in anatomic structures of
taxonomic importance may eventually induce to errors in the species
identification. The aim of this study is to report the finding of
anomalies in male terminalia of Mi. quinquefer captured in a primary
forest in Itatí department, situated on the left coast of Parana River to
75 km from Corrientes city where patients of (ATL) inhabited. Sand
flies were collected, with automatic light traps (CDC type) and using
manual captures. The collections were conducted during four months
of 2010 (April, September, November and December) and 2011
(January, February, Mars and April). The classification system used
was that of Young y Duncan (1994) and Galati (2003). A total of 3,563
Mi. quinquefer was captured (1,520 males and 2,043 females). In
2010 643 males were captured and asymmetrical anomalies in the
number of spines of style were observed in 0.9% of the collected
males. Herein, we described the morphological anomalies Two
males had an supplementary spine implanted in the middle in a style
Four males had a supplementary terminal spine on one style. As these
structures are paired, examination of the corresponding structure on
the opposite side of the body normally permits correct identification.
It is important to publish this finding, because new taxa can be
described from anomalous specimens, leading to confusion among
specialists as to the validity of certain species.
N° 26 (1) 2015
91
THE USE OF DNA BARCODING FOR TAXONOMIC
IDENTIFICATION OF THE IMMATURE STAGES OF
PHLEBOTOMINE SANDFLIES IN OVEJAS, SUCRE
(COLOMBIA)
SAND FLIES (DIPTERA: PSYCHODIDAE) FROM
PERU: REPORT OF NEW RECORDS
Roberto Fernandez, Victor Zorrilla, Liz Espada, Gissella Vásquez &
Frederick Stell
Laura Posada-López1,2; Rafael Vivero1,2; Sandra I Uribe1,2; Luz A.
Acosta1; Andrés Vélez-Mira1; Horacio Cadena1; Luis G Estrada3; Edgar
Ortega3; Eduar Bejarano3; Ivan D Vélez1
NAMRU-6 Lima-Peru.
roberto.fernandez@med.navy.mil
1
PECET-Programa de Estudio y Control de Enfermedades Tropicales, Universidad de
Antioquia, Medellin-Colombia; 2GSM Grupo de sistemática molecular, Universidad
Nacional sede Medellin-Colombia; 3Grupo de Investigaciones Biomédicas,
Universidad de Sucre, Sincelejo, Colombia
lauraposada22@gmail.com
Keywords: Barcode, inmature, natural breeding
Introduction: Phlebotomine sandflies studies in endemic areas of
leishmaniasis, usually only include the collection and identification of
adults, ignoring the importance searching and identifying the
immature stages to confirm the presence of all potential vector
species. This problem arises from the difficulty of finding breeding
sites and the lack of taxonomic keys for immatures. The mitochondrial
cytochrome oxidase I gene (COI) has been successfully used as a
DNA barcode for the molecular identification of insect species of
medical importance, making it ideal for the identification of immature
phlebotomine sandflies. In this study we aimed to validate the use of
the COI gene (5'end) as a tool (barcode) for the identification of
immature phlebotomine sandflies isolated from natural breeding sites.
Methodology: Adult and immature stages of sandflies were collected
in Ovejas, a cutaneous and visceral leishmaniasis foci in the Colombia
Caribbean Coast on 2013. Adults were identified by morphology
using the taxonomic key of Young & Duncan 1994. We extracted total
DNA from phlebotomine sandflies larvae and pupae, amplifying a
product of 700 bp from the COI gene, using the adults order to
compare the sequences (n=17, eight species). We evaluated patterns of
nucleotide variability, calculated the genetic distance K2P (intra and
interspecific), and generated a dendrogram using neighbor joining.
We used our database of sequences obtained from adult Lutzomyia in
Colombia, which consists of 36 species (148 sequences), to verify
identity. Result: We obtained sequences of the 700bp fragments of the
COI mitochondrial gene, from larvae (n=15) and pupae (n=2). The
identity of the immature stages corresponds mainly to Lu. evansi and
Lu. atroclavata, which was determined by comparison with the
sequences of adults previously identified to species by morphological
taxonomy. The percentages of variation of Lu. evansi were 1.7% in
larvae and 2.3% in adults. We found groups with brackets of 99% for
larvae and adults of Lu. evansi and other species identified with adults
(Lu. cayennensis, Lu. atroclavata, Lu. micropyga, Lu. rangeliana, Lu.
gomezi, Lu. shannoni and Lu. dubitans). Conclusion: The COI gene is
a suitable molecular marker for the identification in adults and
immature sand flies. High nucleotide variability in adult and immature
in Lu. evansi in the same location was detected, however the rates of
divergence are within the proposed range for intraspecific difference.
Keywords: Sand flies, Peru
Phlebotomine sand flies are distributed in tropical and sub-tropical
regions of the world, where some species are involved in the
transmission of human pathogens including bacteria, parasites and
arboviruses. In Peru, sand flies are distributed in the occidental slopes
of the Andes as well as in lowland areas in the Amazon Region.
Currently, the New World sand fly fauna comprises 512 species
distributed in many taxonomic groups. Since 1986, we have been
conducting sand fly surveillance in Peru with the goal of
characterizing the sand fly fauna and sand fly species distribution in
different ecological environments, and identifying the sand fly
vectors of leishmaniasis and bartonellosis. Using internal and
external morphological characteristics of phlebotomine sand flies we
have identified 188 species distributed in two genera, Lutzomyia and
Brumptomyia, and a total of 13 subgenera, 8 species group and 5 nongrouped species. In this report we include 15 new records for Peru,
all found in the Amazon Region: L. monstruosa, L. pinottii, L.
falcata, L. araracuarensis, L. cultellata, L. spathotrichia, L.
ferreirana, L. christenseni, L. duncanae, L. beniensis, L. sp de
Tarapoto, L. gantieri, L. guilvardae, B. beapertuyi, and B. leopoldoi.
N° 26 (1) 2015
92
FIRST RECORD OF THE GENUS PINTOMYIA COSTA
LIMA, 1932 (DIPTERA PHLEBOTOMINAE) IN THE
NORTHWEST OF ARGENTINA
MORPHOMETRIC STUDY OF LUTZOMYIA
L O N G I PA L P I S P O P U L AT I O N S ( D I P T E R A :
PSYCHODIDAE) COLLECTED IN URBAN AND
ENVIRONMENTAL PROTECTED AREAS IN MINAS
GERAIS, BRAZIL
Fuenzalida AD1,2,3, Direni Mancini JM1,2, Salomon OD1,2,4, Quintana
MG1,2,3,4
1
Reis, A. S., Saraiva, L., Gontijo, C.M.F., Andrade Filho, J. D.
Instituto Nacional de Medicina Tropical – MSN, 2 REDILA, 3 Instituto Superior de
Entomología – UNT, 4 CONICET.
Centro de Pesquisas René Rachou/FIOCRUZ
fuenzalidadenise@gmail.com
alanna.reis@cpqrr.fiocruz.br
Keywords: Pintomyia, Jujuy, Tucumán, Argentina
Keywords: Lutzomyia longipalpis, Leishamaniasis, Taxonomy,
Phlebotominae
Currently 30 species of sand flies from 13 genera have been recorded
in Argentina. Whithin the genus Pintomyia, seven species have been
recorded in four Eastern provinces of the country: Pintomyia
bianchigalatiae (Andrade-Filho, Aguiar, Dias & Falcão, 1999), Pi.
damascenoi (Mangabeira, 1941), Pi. fischeri (Pinto, 1926), Pi.
misionensis (Castro, 1959), Pi. monticola (Costa Lima, 1932), Pi.
pessoai (Coutinho & Barretto, 1940) and Pi. torresi (Le Pont &
Desjeux, 1991). As part of research project of which objective is to
validate distribution maps of different Phlebotominae species, capture
were performed in areas with no previous records, between March and
May 2013. In Jujuy (24º 11' 04.13”, 65º 18' 02.54”) six sample sites
were select, and eight sites in Tucumán province (26º 48' 28.69”, 65º
13' 09.09”), both corresponding to the transition area of Yungas rain
forest and dry Chaco. Captures were made with mini CDC like lighttraps. In Jujuy province a total of 32 individuals were caught, which
belong to Migonemyia migonei (França, 1920) (47%), Nyssomyia
neivai (Pinto, 1926) (25%), Pintomyia sp. (16%), y Evandromyia
cortelezzii (Brèthes, 1923) (12%). In Tucumán 270 individuals were
collected Mg. migonei (77%), Pi. sp. (18%), Ev. cortelezzi (5%). The
presence of Pintomyia is recorded for the first time in to northwest
province of Argentina. Pintomyia torresi were recorded to date, only
in the province of Chaco; being aware of the relationship of this
species feeding on bats, it is noted that the capture individuals belong
to site in the presences of bats, and in the case of Tucumán province,
the sample area presents the third greatest colony of insectivorous bats
of the South America (Tadarida braziliensis). The identified samples
of this work, belonging to genus Pintomyia (44 female) and (11 males)
were not identified a specific level, although the studied in
morphologic characters allow it to be included in the genus and
subgenus Pintomyia (Pifanomyia). The morphologic similarities to
Pi. torresi in the case of the female genitalia characters , but they are
different in the cibarium characters; in the case of males related to this
female the differences are more precise, especially in structures
associated to the paramere. Due to all above mentioned, a deeper
studies will extended in order to determine whether this is a new
species for Science.
Leishmaniasis are a serious public health problem in Brazil. Belo
Horizonte and its metropolitan region present high incidence and
mortality due to Visceral Leishmaniases. Lutzomyia longipalpis is the
main vector of Leishmania infantum in the country. In the present
study morphological aspects of 30 specimens from an urban area,
located in Belo Horizonte (Venda Nova – Sanitary District) and 30
specimens from an environmental protected area, the Sumidouro
State Park distant 50 km from Belo Horizonte were analyzed.
Samplings were conducted from August 2011 to August 2013, using
CDC traps during three consecutive days. The specimens were
prepared, assembled and submitted to morphometric measurements.
The comparisons between males from the two study populations,
demonstrated statistical significant differences in mean measures of
six characters (length of Labrum-epipharynx, P1, P4, P5, length
gamma and length of lateral lobe) from the 52 analyzed. To the
females, significant differences were observed in the mean of nine
characters (AIII, AXVI, AIV, P1, P3, P4, length gamma and length of
spermathecae) of the 46 analyzed. Comparing the variance of the two
groups the results indicate that the population of the Sumidouro State
Park is more homogeneous, while the population of Venda Nova is
more heterogeneous taking into account the morphometric
measurements. Lu. longipalpis demonstrates ability of adapting to
different habitats and the highest morphological variability of the
specimens of the species in the urban area may be related to this
behavioral plasticity.
N° 26 (1) 2015
SAND FLIES CAPTURES AND IDENTIFICATION OF
LEISHMANIA SUBGENUS IN GIEMSA-STAINED
SLIDES OF PATIENTS FROM FIVE LOCALITIES OF
SALTA, ARGENTINA.
PHLEBOTOMINAE BIONOMICS IN THE HUMID
CHACO BIO-REGION. CHACO, ARGENTINA
Almazán María Cristina1, Gil José F3, Chanampa Mariana M3, Barroso
Paola A2, Nasser Julio R1, 4, Copa Griselda N1, Oreste F4, Castillo G4
1
Instituto de Investigación de Enfermedades Tropicales, UNSa-Orán; 2Instituto de
Patología Experimental, CONICET-UNSa; 3Instituto de Investigaciones en Energía No
Convencional, UNSa; 4Cátedra de Química Biológica, Facultad de Ciencias Naturales
UNSa.
cristina.almazan90@gmail.com
Keywords: Lutzomyia, Leishmania, PCR, RFLP
Leishmaniasis is a disease which is endemic in 88 countries and in
Argentina remains endemic in 9 provinces. It is caused by several
species of flagellates parasites of the genus Leishmania. The wide
spectrum of clinical manifestations depends on Leishmania spp and
the host inmuno-response. The parasite is transmitted by sand flies of
Family Psychodidae, Subfamily Phlebotominae. In Argentina, many
sand flies have been described, being the most important and
suspected vector Nyssomyia neivai because it showed natural
infection with Leishmania parasites. In our country some Leishmania
species were isolated Leishmania (Viania) braziliensis, Leishmania
(Leishmania) amazonensis, L. (V.) guyanensis, L. (V.) panamensis
associated to tegumentary leishmaniasis (TL); and L. (L.) infantum
associated to visceral leishmaniasis. The aim of this work was to
explore the sand flies presence from five localities of the north of
Salta. Besides, patient samples from these areas, were analyzed in
order to identify the causal agent. Sand flies were collected during one
or six nights with CDC light traps. Traps were placed a meter above the
soil and left since 18 pm to 7 am. Species identification of all collected
sand flies was made by morphology of spermatheca and cibarium or
external genitalia. Leishmania subgenus determination was carried
out by PCR-RFLP assay. Seventy-six giemsa-stained samples
diagnosed for TL in 2002 at Instituto de Investigación de
Enfermedades Tropicales located in Orán, Salta were used. DNA
extractions were made by a lysis buffer. The PCR primers L5.8S
5 ' T G ATA C C A C T TAT C G C A C T T 3 ' a n d L I T S R n
5'CTGGATCATTTTCCGATG3' were used. For RFLP, amplicons
were digested with HaeIII enzyme. We captured 1352 phebotomines,
of which Ny. neivai represent the 76%, Migonemyia migonei 12%,
Evandromyia cortelezzii complex 11% and Psathyromyia shanonni
1%. Ny. neivai was present in all localities studied. By PCR-RFLP,
Viannia subgenus was the only identified. Regarding geographical
cases distribution, 97% of them were from Orán, being this area the
site with more cases and highest sand flies amount. Three sandflies
species found in this study are considered as potential cutaneous
leishmaniasis vectors, particularly of Viannia subgenus parasites. We
propose PCR-RFLP in order to enhance the traditional diagnostic just
because knowing the leishmaniasis causal agents would improve the
treatment assignation and we suggest sandflies surveillance in these
localities.
93
Szelag Enrique A1,2, Rosa Juan R.1, Parras Matias A.1,2, Salomón Oscar
D.2
REDILA 1Instituto de Medicina Regional (Universidad Nacional del Nordeste).
Instituto Nacional de Medicina Tropical.
2
szelag_enrique@yahoo.com.ar
Keywords: Tegumentary Leishmaniasis, Nyssomyia neivai,
Migonemyia migonei
In Argentina, Tegumentary Leishmaniasis (TL) was first recorded in
1916. Actually is considered endemic in 9 provinces from north-east
to north-west of the country. Urban Visceral Leishmaniasis (VL)
produced the first autochthonous human case in Argentina in the city
of Posadas in 2006. Currently, transmission is recorded in Misiones,
Corrientes, Santiago del Estero, Formosa and Salta. In order to
characterize the distribution in space and time of potential vectors of
TL and VL; during the period October 2010 - September 2012
samplings were carried out in three areas of the Chaco province:
Resistencia, Margarita Belén and Colonia Benítez. In each area an
inhabited dwelling was selected, and CDC light traps were installed
monthly during one single night from 7pm to 7am next day in
domicile, Peri-domicile and forest. Data of Relative Humidity, High
and Low Temperature, were recorded. The Phlebotominae captured
were clarified and identified according to Galati (2003). A total of
1499 Phlebotominae was captured in this study. Eight species were
recorded: Brumptomyia brumpti, Br. avellari, Evandromyia
cortelezzii, Ev. sallesi, Migonemyia migonei, Sciopemyia sordelli,
Nyssomyia neivai, Psathyromyia shannoni. Predominant species
were Mg. migonei(41.03%) followed by Ny. neivai(38.96%) and
complex cortelezzii(9.54%). The greater abundance of
Phlebotominae was recorded in Forest(n=1003), followed by Peridomicile(n=423) and domicile(n=73). Mg. migonei predominated in
summer with an average temperature of 28.09°C and relative
humidity of 61%, while in spring the most frequent species was Ny.
neivai with averages of 22.5 ºC and 58% RH. The lower abundance of
Phlebotominae was recorded between June and July. The increased
frequency of these species in warm and dry months should be
considered when proposing control measure. In conclusion, the
presence of competent TL vectors as Ny. neivai and Mg. migonei (also
a potential vector of VL) in the forest and in lower proportion in
peridomestic environments, suggested for the region a risk of
transmission of leishmaniases, mainly due to human activities related
to the forest, but also by periodical colonization of the peridomestic
areas with population of vectors from the forest. Further studies
should determine for TL the role of Mg. migonei, the main vector in
the Dry Chaco region, as an opportunistic vector in hot-spot
transmission, and its interaction with Ny. neivai the main vector in the
Humid Chaco region of Argentina.
N° 26 (1) 2015
DISPERSION OF LUTZOMYIA LONGIPALPIS IN THE
CHACO PROVINCE, ARGENTINA
MONTHLY DISTRIBUTION OF LUTZOMYIA
LONGIPALPIS IN AN ENDEMIC AREA OF VISCERAL
LEISHMANIASIS IN SÃO PAULO STATE, BRAZIL
Szelag Enrique A.1,2, Parras Matias A. 1,2, Rosa Juan R. 1, Salomón Oscar
D.2
REDILA 1Instituto de Medicina Regional (Universidad Nacional del Nordeste) –
Instituto Nacional de Medicina Tropical.
2
szelag_enrique@yahoo.com.ar
Keywords: VL - Visceral Leishmaniasis - Migonemayia migonei
In Argentina Lutzomyia longipalpis, the main vector of Leishmania
infantum, was recorded for the first time in 2004 in Formosa province.
In the following years, the vector spread to the south and west of the
country being recorded in the provinces of Misiones (2006),
Corrientes (2009), Chaco, Entre Ríos (2010) and Salta (2013). Given
this fact, the aim of this work was monitoring the spread of Lu.
longipalpis in a receptive area of Visceral Leishmaniasis (VL)
transmission, in the Chaco province. Captures of Phlebotominae were
made in the city of Resistencia (27°27´S 58°59'W), based on a
stratified systematic sampling method. From November 2010 to May
2012, four strata were sampled once every season, except in winter.
The samples were selected by listing and numbering the
neighborhoods of each stratum and randomly, selecting 12
neighborhood per season. In each neighborhood, the house considered
the “worst case epidemiological scenario” was selected as sample site.
At each sample site, insects were captured by light CDC traps located
at 1.5 m, above the animal shelter (kennel, henhouse, etc.) in the
courtyard of the house, during one single night (from 17.00 to 9.00 h).
Urban localities near Resistencia were also selected as sentinel sites
sampled one per season, to monitor any eventual spread from
Resistencia. These sentinel sites included the villages of Margarita
Belén (27°15'S- 58°58'W), Isla del Cerrito (27º17'S- 58°37'W) and
Barrio de los Pescadores (27°27'S- 58°51'W).During the study period,
14 out of 85 traps installed had at least one phlebotomine, obtaining a
total of 38 Phlebotominae distributed in three locations: Resistencia,
Margarita Belen and Barrio de Pescadores. The species recorded were
Lu. longipalpis (13.2%), Migonemyia migonei (15.8%), Nyssomyia
neivai (39.5%) and cortelezzii complex (31.6%), represented by
Evandromyia sallesi and Ev. cortelezzii. In this monitoring Lu.
longipalpis was absent in the city of Resistencia, and its presence was
restricted to Barrio de los Pescadores, the closest area to the city of
Corrientes (with confirmed presence of the vector, VL human and
canine cases). In conclusion, this monitoring of Lu. longipalpis
showed that the incipient colonization of the VL vector observed in
2010 with captures of low relative abundance in the city of
Resistencia, was not followed by continuous installation of vector
populations and expansion of their spatial distribution as in other
urban centers of Argentina.
94
Fredy Galvis Ovallos1, Anaiá da Paixão Sevá2, Estela Galucci2, Rodrigo
Martins Soares3, Fernando Ferreira3, Eunice Aparecida Bianchi Galati4
1
Postgraduate Program in Public Health, School of Public Health, University of São
Paulo- USP, Brazil. 2Postgraduate Program in Experimental Epidemiology of
Zoonoses, School of Veterinary Medicine and Animal Science, University of São
Paulo-USP, Brazil. 3Department of Preventive Veterinary Medicine and Animal
Health, School of Veterinary Medicine and Animal Science, University of São PauloUSP, Brazil. 4Department of Epidemiology, School of Public Health, University of São
Paulo – USP, São Paulo, SP, Brazil
galvisfregao@gmail.com
Keywords: Visceral leishmaniasis, São Paulo, Lutzomyia
longipalpis
Visceral leishmaniasis represents a great challenge to public health in
Brazil in view of the great, widespread incidence of both human and
canine infection. The area affected by visceral leishmaniasis in São
Paulo state has been expanding since 1998. Human and canine cases
have been registered in 105 municipalities, the presence of the main
vector, Lutzomyia longipalpis, having been registered in the urban
area of 148 municipalities. This study sought to describe the
Phlebotominae fauna in the urban area of Panorama municipality - SP
and the monthly frequency of Lu. longipalpis. The sandfly captures
were made from July 2012 to April 2014 with automatic light traps.
Two traps were installed in the intra and peridomicile of seven
dwellings, selected for the presence of an environment propitious to
sandflies. Lutzomyia longipalpis predominated (94.3%), followed by
Nyssomyia neivai (3.8%), Evandromyia lenti, Ev. cortellezzi and
Brumptomyia brumpti. A total of 4,115 specimens of Lu. longipalpis
were captured with a 3:1 male/female ratio. The highest average of
sandflies/night/trap was obtained in the rainy season (NovemberMarch) and the highest frequencies of males and females were
observed in March 2013. A positive and significant association was
detected between the number of sandflies/trap/night and the total
monthly rainfall (Spearman r= 0.48). Vector density constitutes the
main parameter related to the dynamic transmission of vector borne
diseases and the knowledge of the monthly distribution of the vector
could contribute to the design and planning of control measures as
well as to other strategies for the prevention of the disease. Our results
suggest that the risk of transmission of L. infantum chagasi in
Panorama city increases in the rainy season and that vector control
activities should be undertaken especially in that period. Financial
support: FAPESP – Process no. 2012/03751-4 and no. 2012/01442-4
N° 26 (1) 2015
FREQUENCY OF LUTZOMYIA LONGIPALPIS IN
HENHOUSES IN AN AMERICAN VISCERAL
LEISHMANIASIS ENDEMIC AREA OF SÃO PAULO
STATE
MICROPYGOMYIA QUINQUEFER (DYAR, 1929)
(DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE): A
PUCTATIVE VECTOR OF LEISHMANIA SPP.?
1
2
2
Fredy Galvis Ovallos , Anaiá da Paixão Sevá , Estela Galucci , Rodrigo
Martins Soares3, Fernando Ferreira3, Eunice Aparecida Bianchi Galati4
1.
Postgraduate Program in Public Health, School of Public Health, University of São
Paulo- USP, Brazil. 2.Postgraduate Program in Experimental Epidemiology of
Zoonoses, School of Veterinary Medicine and Animal Science, University of São PauloUSP, Brazil. 3.Department of Preventive Veterinary Medicine and Animal Health,
School of Veterinary Medicine and Animal Science, University of São Paulo-USP,
Brazil. 4.Department of Epidemiology, School of Public Health, University of São Paulo
– USP, São Paulo, SP, Brazil
95
Cristian Ferreira Souza1; Paula Dias Bevilacqua2; José Dilermando
Andrade Filho3; Reginaldo Peçanha Brazil4; Patrícia Flávia Quaresma5
1
Programa de Pós-Graduação em Biologia Parasitária- Instituto Oswaldo Cruz
Fundação Oswaldo Cruz–Manguinhos, Rio de Janeiro–RJ. Brazil. 2Laboratório de
Leishmanioses-Centro de Pesquisas René Rachou/Fiocruz, Belo Horizonte–MG.
Brazil. 3Centro de Referência Nacional e Internacional para Flebotomíneos,
Centro de Pesquisas René Rachou/Fiocruz, Belo Horizonte–MG. Brazil.)
4
Laboratório de Doenças Parasitarias-Instituto Oswaldo Cruz - Fundação Oswaldo
Cruz–Manguinhos-Rio de Janeiro–RJ. Brazil. 5Departamento de Veterinária Universidade Federal de Viçosa-Campus Universitário-Viçosa–MG. Brazil
biominas2004@yahoo.com.br
Keywords: Henhouses, Visceral leishmaniasis, Panorama-SP
American visceral leishmaniasis (AVL), caused by Leishmania
infantum chagasi and transmitted mainly by Lutzomyia longipalpis, is
widespread in Brazil and in recent decades has expanded to new
regions of the country. This study aimed to describe the effect of the
removal of feces from henhouses on the density of AVL in São Paulo
State. The sandfly captures were made in eight dwellings: three with
henhouses from which the chicken feces were removed (FR+), three
with henhouses without the removal of the chicken feces (FR-) and
two residences without henhouses (Control - C). The captures were
undertaken with two automatic light traps in each residence between
July 2012 and June 2013. A total of 2,238 males and 746 females of Lu.
longipalpis were captured. 76.6% of the insects were captured in the
FR- houses, followed by C (14.3%) and FR+ (9.0%). Our results
suggest that the removal of the feces from henhouses may have
contributed to the decreases in Lu. longipalpis frequency, because of
the removal of the organic matter necessary for the establishment of
breeding sites. However, this result should be carefully analyzed
because the sandfly frequency is also affected by other environmental
characteristics such as vegetation cover and the availability of hosts.
Henhouses are recognized as risk factors for AVL transmission,
nevertheless the possibility of their elimination from urban areas
constitutes a challenge in view of the fact that their presence is
associated with socioeconomic and cultural factors. Further studies
should be undertaken covering a larger sample of households with
henhouses so as better to assess the effect of feces on the frequency of
this sandfly species. Financial Support: FAPESP - Process no.
2012/03751-4 and no 2012/01442-4.
Keywords: Leishmaniasis, sand fly, trypanosomatid, natural
infection
Micropygomyia quinquefer is a species of sand flies which, up to now,
is not suspected of participating in the transmission of any
Leishmania cycle. This is mainly due to the fact the species integrate a
group of sand flies, which according to the literature, feeds
exclusively on cold-blooded animals. This study aimed to verify the
presence of natural infection in females of Mi. quinquefer captured in
the urban area of the municipality of Timoteo, Minas Gerais, Brazil.
Sand fly collections were performed in the city with Falcao light
traps, exposed monthly for two consecutive nights during the period
of November/2009 to October/2010. Captured females who had no
signs of blood meal in their abdomen, were separated into pools of up
to 10 females of the same species and used in molecular assays for the
detection of natural infection by Leishmania spp. by PCR and RFLP
ITS1. 1,958 sandflies were collected, with Nyssomyiawhitmani the
most captured species (66.5%) and Mi. quinquefer the second
(15.4%). Among the sand fly species of Mi. quinquefer, 251 (83.1%)
females were divided into 91 pools for molecular analysis, 23
(25.3%) were positive for Leishmania spp. Infection by PCR of ITS1
and 8 (8.8%) were positive for L. braziliensis by RFLP of ITS1. This
is the third record of Mi. quinquefer with Leishmania DNA in Brazil.
This finding is of particular ecological significance; since it is
believed that females of Mi. quinquefer feed on cold-blooded animals
and cannot has a role in the transmission cycle of Leishmania sp.
Another interesting detail was the identification of a large number of
positives pools for L. braziliensis. Hence that L. braziliensis only
infects mammalian species, these results suggest that Mi. quinquefer
can feed on warm-blooded animals, or even that molecular markers
used are amplifying DNA fragment from other trypanosomatid, not
yet described by science. Although it cannot establish this species as a
vector of L. braziliensis, we cannot ignore that it was the second most
frequently captured species in the study area, being characterized by
the occurrence of human cases. This shows the necessity of other
studies to understand the participation of this sand-fly species as a
vector in eco-pidemiology of leishmaniasis in the area.
N° 26 (1) 2015
CANINE ANTIBODY RESPONSE TO PHLEBOTOMUS
PERNICIOSUS AND PHLEBOTOMUS ORIENTALIS
BITES.
EPIDEMIOLOGICAL ASPECTS OF VECTOR,
PARASITE, AND DOMESTIC RESERVOIR IN AREA
WITH NO REPORTED HUMAN CASES OF VISCERAL
LEISHMANIASIS IN BRAZIL
Tereza Kratochvílová1, Tatiana Košťálová1, Michaela Kindlová1, Petra
Sumová1, Michal Šíma1, Iva Rohoušová1, Marina Gramiccia2, Luigi
Gradoni2, Asrat Hailu3, Gad Baneth4, Alon Warburg5, Petr Volf1
1
Department of Parasitology, Faculty of Science, Charles University in Prague,
Vinicna 7, 128 44 Prague 2, Czech Republic; 2 Unit of Vector-borne Diseases and
International Health, MIPI Department, Istituto, Superiore di Sanità, Rome, Italy; 3
Department of Microbiology, Immunology & Parasitology, Faculty of Medicine, Addis
Ababa University, P.O. Box 9086, Addis Ababa, Ethiopia; 4 School of Veterinary
Medicine, Hebrew University, P.O. Box 12, Rehovot 76100, Israel; 5 Department of
Parasitology, The Kuvin Centre for the Study of Infectious and Tropical Diseases,
Hadassah Medical School, The Hebrew University of Jerusalem, Jerusalem, Israel.
96
Fabiana de Oliveira Lara-Silva1, Érika Monteiro Michalsky1, Consuelo
Latorre Fortes-Dias2, Vanessa de Oliveira Pires Fiuza3, José Eduardo
Marques Pessanha3, Shara Regina da Silva1, Daniel Moreira de Avelar1,
Maiara Alves Silva1, Ana Cristina Vianna Mariano da Rocha Lima1,
Ailton Junior Antunes da Costa1, George Luiz Lins Machado-Coelho4,
Edelberto Santos Dias1
1
Laboratório de Leishmanioses, Centro de Pesquisas René Rachou/Fiocruz, Belo
Horizonte, Minas Gerais, Brazil; 2Diretoria de Pesquisa e Desenvolvimento,
Fundação Ezequiel Dias, Belo Horizonte, Minas Gerais, Brazil; 3Secretaria Municipal
de Saúde de Belo Horizonte, SMSA/PBH, Brazil; 4Universidade Federal de Ouro
Preto, Minas Gerais, Brazil
tereza.kratochvilova@natur.cuni.cz
flara@cpqrr.fiocruz.br
Keywords: P. perniciosus, P. orientalis, antibody, canine
In the Mediterranean area, dogs serve as the main reservoir host of
Leishmania infantum, the causative agent of zoonotic visceral
leishmaniasis (VL). In East Africa, the role of dogs in the
epidemiology of VL caused by L. donovani is not clear. In both areas,
however, the disease is transmitted by sand flies of the subgenus
Larroussius; Phlebotomus perniciosus and P. orientalis are major
vectors of VL in the Mediterranean and East Africa, respectively.
Frequent contact of the sand fly vector with the host is a prerequisite
for transmission and therefore we were interested to learn more about
exposure of dogs to these sand fly species. Sand fly saliva inoculated
into the feeding site during bloodmeal intake elicits an antibody
response that can be utilized as a specific marker of exposure. The
intensity of exposure correlates with the antibody response against
saliva and may serve as an epidemiological tool for estimating the
vector bites and also for the risk of Leishmania transmission.
However, since the use of whole salivary homogenate is limited by
maintenance of sand fly colonies and laborious dissection of their
salivary glands, recombinant salivary proteins have recently been
tested to replace salivary gland homogenate in serological surveys. In
our study we identified major salivary antigens of P. perniciosus and P.
orientalis by western blotting and mass spectrometry (MALDI-TOF)
and tested them as potential markers of dog exposure to these sand fly
vectors. The two most promising P. perniciosus recombinant proteins,
43-kDa yellow-related protein (rSP03B) and 35.5-kDa apyrase
(rSP01) were tested by ELISA with more than 750 canine sera
collected in southern Italy, an area endemic for VL. Similarly, we
studied reaction of P. orientalis salivary proteins with canine sera
collected in a L. donovani focus in Ethiopia. Canine sera originating
from dogs living in non-endemic countries were used for negative
control. Experiments showed that dogs from both endemic areas had
significantly increased IgG levels against saliva of the local vector.
Moreover, the results proved that recombinant salivary proteins of
sand flies can fully replace salivary gland homogenate in ELISA tests.
Keywords: Phlebotominae, Lutzomyia longipalpis, Leishmania
infantum, natural infection
Visceral leishmaniasis (VL) is a neglected disease of great
importance in Public Health. In the Americas, about 97% of the
human cases of VL occur in Brazil. In the last few years, the disease
has been expanding to medium and large size cities and the actions
taken by the Brazilian Program for Surveillance and Control of VL
are more intense where the epidemiological transmission risk (ETR)
is higher. Belo Horizonte, the sixth most populous Brazilian city, is
among the state capitals with intense ETR of VL. The ETR, however,
varies depending on the urban district analyzed. We focused our twoyear study in districts of Belo Horizonte with no reported human
cases of VL, so far, in an effort to draw a current epidemiological
profile of the local situation concerning vector, parasite, and domestic
reservoir. Lutzomyia longipalpis, the main vector of VL in Brazil, was
highly abundant, mostly in the peri-domiciles. The species comprised
96.5% of all the phlebotomine sand flies captured, with increased
numbers after rainfall peaks. DNA of Leishmania infantum, the
etiological agent of VL, was detected in 88.5% of the Lu. longipalpis
pool samples. Le. infantum DNA was also present in the cortelezzii
complex and in Lutzomyia lloydi. DNA of Leishmania braziliensis,
etiological agent of the cutaneous leishmaniasis, was also identified
in Lu. longipalpis. Among the 1,410 dogs serologically tested for VL,
3.6% were positive. These dogs were additionally submitted to two
parasite-based and one molecular-based diagnostic tests. The main
result was that Leishmania infection was confirmed in all of them
when the spleen tissue was used as DNA source in the molecular test.
Moreover, the infecting parasite was genotyped as Le. infantum in all
of them.
We concluded that the parasite–vector and the
parasite–domestic reservoir interfaces are very active in those
districts, even in the absence of reported human cases of VL,
therefore providing favorable conditions to local outbreaks in a short
period of time. Financial support: FAPEMIG/FIOCRUZ/SMSBH.
N° 26 (1) 2015
SANDFLIES FAUNA SURVEY AND DNA DETECTION
OF LEISHMANIA INFANTUM IN LUTZOMYIA
LONGIPALPIS CAPTURED IN THE CITY OF SABARA,
P H L E B O TO M I N E S A N D F L I E S ( D I P T E R A :
PSYCHODIDAE) IN CUTANEOUS LEISHMANIASIS
ENDEMIC AREAS OF THREE MUNICIPALITIES OF
THE STATE OF PARÁ, BRAZIL
Érika Monteiro Michalsky1, Josiane Valadão Lopes1, Fabiana Oliveira
Lara E Silva1, Maiara Alves Silva1, Ana Cristina Vianna Mariano Da
Rocha Lima1, João Carlos França-Silva2, Fabiana Rodrigues Da Paixão1,
Ailton Junior Antunes Da Costa1 & Edelberto Santos Dias1
Ana de Nazaré Martins da Silva; Edna Aoba Yassui Ishikawa; Mônica
Fadul; Thiago Vasconcelos dos Santos; Yago Costa Vasconcelos dos
Santos; Patrick Thiago Carvalho de Araújo e Nathália Queiroz Santos
1
2
Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, Belo Horizonte, Brasil
Universidade Federal de Minas Gerais, Belo Horizonte, Brasil
97
Núcleo de Medicina Tropical, Universidade Federal do Pará, Brazil
ishikawa@ufpa.br
erika@cpqrr.fiocruz.br
Keywords: Lutzomyia, sandfly, cutaneous leishmaniasis, Brazil
Keywords: Leishmania infantum, Lutzomyia longipalpis, natural
infection, Sabará
Visceral leishmaniasis (VL) is a neglected disease of great importance
in Public Health, which is present in 65 countries around the world. In
Brazil, VL urbanization is a fact, with increasing numbers of human
cases and high prevalence of the canine cases in large size cities. In the
New World, Leishmania infantum is the etiologic agent and its
transmission occurs through the bite of the females of Lutzomyia
genus sandflies. LV is distributed in the city of Sabara, characterizing
the region as an endemic area. The aim of this work was to survey the
city's sandflies fauna, as well as study the behavior of the species in
domestic environments and detect natural infection of Lutzomyia
longipalpis by Leishmania spp. The entomological captures were
performed in 8 districts of the city (Alvorada, Novo Alvorada,
Alvorada Velho, Bom Retiro, Ana Lúcia, Rio Negro, Nova Vista e
Casa Branca), during the period of 01/2011 to 12/2012. These were
made monthly for three consecutive days using the HP type light traps
installed inside and outside homes. The males collected were stored in
70% ethanol for taxonomic identification and females were dissected
and analyzed by Nested /PCR, afterwards, DNA sequencing was made
for identification of circulating Leishmania species. 519 specimens
were captured and 4 different species were identified. The species
distribution was: L. longipalpis (95.0%), L. cortelezzii (2.3%), L.
whitmani (1.5%) and L. intermedia (0.2%). Regarding the behavior of
the species, 13% were caught inside and 87% outside the homes. Of
the 9 L. longipalpis "pools" analyzed, the 353 bp fragment typical of
Leishmania spp. was observed in 3 of them, keeping the minimum
infection rate around 33%. Of the 3 female L. longipalpis "pools"
positive for Leishmania, were made DNA sequencing and sequences
alignment and we observed that the Leishmania species circulating in
the vectors of city of Sabara is L. infantum. The sandflies fauna
showed leishmaniasis vector species, both visceral and cutaneous.
The large number of captured L. longipalpis (95%) and a high
infection rate by L. infantum (33%), associated with high canine
positivity (4.25%) and the occurrence of human cases of VL,
demonstrate that the cycle of the disease transmission is well
established in the studied urban area. Financial support:
FAPEMIG/FIOCRUZ/CNPq/SMS-Sabará
The number of cutaneous leishmaniasis cases (CL) notified from
2010 to 2012 has demonstrated a fast expansion in state of Pará,
Brazil, and the Healthy Department rated 26.57% of the
municipalities as very high transmission area. The aim of this work
was to study the fauna and to detect natural infection by Leishmania
spp in sand flies captured in six communities with or without recent
record of CL in human: Villages Betel and Paraíso (Dom Elizeu);
Settlements Rossi Gabriel and Nova Vida (Ulianópolis); Villages
Água Branca and Santa Helena (Rondon do Pará). Sand flies were
captured using CDC light traps placed in peridomicile areas. The
traps were installed 1.5 m above ground level at 6:00 pm and removed
the next day at 6:00 am. The taxonomic identification of each
specimen determined was based on Young and Duncan (1994) and all
females were selected for PCR analysis. In 2011, a total of 229
specimens were captured: Dom Elizeu: Lutzomyia flaviscutellata
(35.21%), Lu. evandroi (18.31%), Lu. whitmani (11.27%), Lu. choti
(9.86%), Lu. davisi (9.86%), Lu. sericea (1.41%), Lu. termithophila
(1.41%), Lu. bacula (1.41%), Lu. sordellii (1.41%), Lu. hirsusta
hirsuta (1.41%); Rondon do Pará: Lu. evandroi (60.0%), Lu.
triacantha (13.34%), Lu. shannoni (6.67%), Lu. gomezi (6.67%), Lu.
dasypodogeton (3.33%), Lu. evandroi (3.33%), Lu. spinosa (3.33%),
Lu. trinidadensis (3.33%), and Lu. whitmani (3.33%); Ulianópolis:
Lu. triacantha (42.19%), Lu. hermanlenti/dreisbachi (23.47%), Lu.
evandroi (9.37%), Lu. wellcomei/complexa (7.037%), Lu.
flaviscutellata (5.47%), Lu. umbratilis (2.34%), Lu. whitmani
(1.56%), Lu. gomezi (0.78%), Lu. longipalpis (0.78%), Lu. shannoni
(0.78%), Lu. trinidadensis (0.78%), Lu. ubiquitalis (0.78%), Lu.
davisi (0.78%). None of the female sand flies were found infected by
Leishmania spp. Despite the low number of specimens captured, Lu.
flaviscutellata, vector of L. (L.) amazonensis was the predominant
species in Dom Elizeu. In Ulianópolis, among medical interest
species, Lu. wellcomei/complexa the main vector of L.(V.)
braziliensis in the region, was the most prevalent, followed by Lu.
flaviscutellata. The presence of sand fly fauna as recorded
demonstrates the need for epidemiological surveillance of CL in these
human settlements, many of them located near the forest areas.
Supported by MS/CNPq/FAPESPA and SESPA.
N° 26 (1) 2015
ID 78-P Ecology & Epidemiololgy
PRELIMINARY STUDY OF SANDFLIES CAPTURED
IN THE CITY OF MACEIÓ-ALAGOAS-BRAZIL
EDUCATION: DEMOGRAPHIC AND SOCIAL
IMPACTS ON THE RATE OF INCIDENCE OF
VISCERAL LEISHMANIASIS IN THE STATE OF
TOCANTINS, BRAZIL (2010)
Ana Lúcia Moreira Macena, Cláudia Cavalcante De Matos Rodarte,
Maria Elisabel Tavares Da Silva, Carlos Fernando Rocha Dos Santos,
Magliones Carneiro Lima, Grimoaldo Braga Da Rocha Neto
98
Secretaria de Saúde Maceió-Alagoas-Brasil
Ana Flávia de Morais Oliveira, Hebert Lima Batista, Keily Cristiny
Azevedo Leite, Helio Márcio Nunes Lacerda
cfr_santos@hotmail.com
Instituto Federal de Educação, Ciência e Tecnologia do Tocantins
anaflaviamo@ifto.edu.br
Keywords: Sandflies, Maceió, Captured
Keywords: leishmaniasis, social indicators, incidence
American visceral leishmaniasis is an anthropozoonosis whose
etiologic agent is the protozoan Leishmaniachagasi. Transmission
occurs through the bite of the infected female sandfly
Lutzomyialongipalpis, which has been registered both in natural
ecotopes as in rural and urban environments, next to domestic animals
and human habitations (Roach et al. 2004). In Latin America, visceral
leishmaniasis has been described in 12 countries, with 90% of the
cases occurring in Brazil, especially in the Northeast Region
(MS/2006). Maceio is a city known as endemic for visceral
leishmaniasis, the Center for Zoonosis Control of the city develops
routinely entomological investigations aiming to gather both
quantitative and qualitative information about sandflies. To the
execution of the leishmaniasis control program, the sand flies were
captured in a non-selective way by CDC light traps. The sampling was
done where the occurrence of human and/or canines positive cases
were registered, being previously installed around 20 traps per night,
1.5m above ground level, for a period of 12 hours. In the urban areas,
the traps were installed intra and extra household, in a sequence of two
nights; while in the rural areas the same trap distribution and sampling
period were used but an extra trap was added outside the house. In the
year of 2013, 1.125 sandfly specimens were found in 13 locations in
the city of Maceió. Of these, 246 were identified as
Lutzomyialongipalpis; 6 L.migonei; 721 L.whitmani; 31
L.corumbaensis; 04 L.quinquefer; 56 L.chotti; 05 L.sallezi; 01
L.tupinambay; 45 L.intermedia; 03 L.evandroi; 04 L.sp (Pressatia);
and 03 L.spp (lenti/carmelinoi). According to the Ministry of Health
(2006), two species are related to the transmission of the disease in
Brazil: L. longipalpis and L. cruzi.
Visceral leishmaniasis is recognized by the World Health
Organization as one of the most endemic and lethal diseases in the
world, not just because of its deadliness, but also because it is very
difficult to treat. This study aimed to analyze the relationship between
the number of cases of VL and the rate of illiteracy, and the average
per capita income. In addition to evaluating the relationship between
the disease and the number of inhabitants, this study also sought to
describe the incidence of the disease in the state of Tocantins during
2010. A descriptive, analytical and retrospective study was done in
2010, using secondary data from IBGE (demographical and social
data) and DATASUS (death rate) through a sample of positive cases
of VL among Tocantins residents. Microsoft Excel 2010, Tabwin, and
Biostat were used to process the data. The general incidence of this
disease was measured from a sample of 100 thousand inhabitants,
taking into consideration only the cases confirmed by the laboratorial
and clinic/epidemiologic criteria. The population used to generate
these statistics came from the 2010 Demographical Census. These
social indicators were pulled from the IBGE (Instituto Brasileiro de
Geografia e Estatística). The social indicators in the IBGE data was
taken from the median per capita income and the average illiteracy
rate. The Pearson theory was applied in order to understand the
relationship between VL incidence, social variables, and the number
of inhabitants. The statistic was considered significant when the
value of p was less than 0.05. The study was made exclusively with
secondary data. The research demonstrated that the median per capita
income pulled from 89.2% of the municipalities in Tocantins was less
than R$ 500.00 per month. The illiteracy rate was considered
alarming: when 42 (30.2%) of the municipalities showed an illiteracy
rate greater than 20%. Approximately 74 (53.2%) of the
municipalities had a population smaller than five hundred thousand
inhabitants. The majority of LV outbreaks were concentrated within 7
municipalities that affected 38% of the state's total population. The
incidence rate of VL was higher than 4 out of 100,000 inhabitants in
41% of the municipalities. In conclusion, there was a significant
correlation between the incidence of VL, the state's illiteracy rate, and
the state's median per capita income.
N° 26 (1) 2015
99
ECO-EPIDEMIOLOGY OF LEISHMANIASIS IN THE
REGION OF SERRA DO CIPO, A MAJOR TOURISTIC
CENTRE OF MINAS GERAIS
PHLEBOTOMINE SAND FLY FAUNA (DIPTERA:
PSYCHODIDADE)
FROM THE LEISHMANISIS
ENDEMIC FOCI IN MATO GROSSO, BRAZIL
Rosana Silva Lana, Erika Monteiro Michalsky, João Carlos França
Silva, Fabiana de Oliveira Lara e Silva, Consuelo Latorre Fortes Dias,
Juliana Martins & Edelberto S. Dias
Sinara Cristina de Moraes, Alan Lane Melo e Elizabeth Ferreira Rangel
Depto. de Parasitologia/ICB/UFMG, Belo Horizonte- MG
Fundação Oswaldo Cruz - Centro de Pesquisas René Rachou
sinaramoraes@hotmail.com
rosana.lana@cpqrr.fiocruz.br
Keywords: Leishmaniasis, sandflies and Leishmania
Keywords: Leishmaniasis, Leishmania, Epidemioly
The purpose of this study was to evaluate the transmission of
leishmaniasis in Jaboticatubas, Serra do Cipó, tourist region of Minas
Gerais. For this, entomological studies and urban reservoirs of the
disease were conducted. For entomological studies, eight districts
were selected, taking into account the occurrence of human cases, and
light traps HP type were placed in the outdoors in 10 homes for three
consecutive nights per month from May 2012 to April 2013. The sand
flies studied consisted of 3,104 specimens , quite diverse with 17
species of Lutzomyia and two females of the genus Brumptomyia.
With regard to species of medical importance and also numerous,
emphasize the meeting of Lu.whitmani (33.80 %), Lu. intermedia
(14.60%) and Lu. longipalpis (7.96 %). The general rate of natural
infection of sand flies by Leishmania was 3.40%. 249 "pools" of
different sand fly species, among them 32 were positive. To study the
rate of canine infection, census survey among domestic dogs in urban
area was carried out. 1,105 serum samples were tested by the DPP®
and confirmed positive infections by ELISA immunoenzymatic assay,
obtaining the average prevalence of canine 7.78%. Of the 86 positive
dogs, a random sample of 11 was selected , which were necropsied and
samples of biopsy tissue obtained were performed " imprints ", culture
of parasites and PCR. Georeferencing was conducted in urban areas of
all canine and human leishmaniasis cases in the last 6 years, and built
up a map showing the distribution of cases. The PCR of marrow
aspirate showed a higher positivity (63.60%) compared to skin
(9.00%). While the "imprints", the positivity rate of skin was 36.36 %
and 0 % marrow. As for the culture of marrow aspirates, the positivity
was 27.27%. Our results indicate the urbanization of leishmaniasis in
the town due to high vector density and infection by Leishmania,
expressive average rate of prevalence canine infection and recent
records of human cases. Therefore, is important to intensify control
measures in order to prevent the problem from getting worse in the
town.
The Leishmaniases are a group of diseases caused by the protozoa of
the genus Leishmania (Kinetoplastida:Trypanosomatidae) which
affect several orders of mammals, including humans, and are
transmitted by the bite of some species of sandflies
(Diptera:Psychodidae). The Leishmaniasis in Brazil has shown a
large geographic expansion. The aim of this study was to compare the
indoor sand flies, peridomicile with that of outside the home (sites
near forests) by entomological research in the urban area, the extra
urban (adjacent to urban) and wild, in the county of Barra do Garças,
Mato Grosso. The entomological survey was carried out from July
2013 to February 2014 with the use of CDC light traps, monthly
collections were made from 3 consecutive nights around 20 houses,
of which 8 captured were inside. Catches of sandf lies were also held
outside the house and in the wild, one night per month, by trapping
Shannon light traps. The sand fly fauna comprises 32 species
representing two genera (Lutzomyia and Brumtomyia)from 4,181
specimens captured. The most frequent species were Lutzomyia (L.)
longipalpis (46.5%), followed by L. lenti (15.1%), L. (N.) whitmani
(12%), L. carmelinoi (9%), L. (L.) cruzi (7.4%), and other species
were twenty seven (10 %) as L. evandroi, L. acanthopharynx, L (S.)
sordellii, B. brumpti, L. termitophila, L. hermanlenti, L. goiana, L
(P.) davisi, L. longipennis, L. (E.) teratodes, L. (P.) punstigeniculata,
L. oliveirai, L. (P.) christenseni, L.(P.) lutziana, L. brasiliensis, L.
ferreirana, L. peresi, L. walkeri, L. (N.) antunesi, L. aragaoi, L. (N.)
flaviscutellata, L. (P.) shannoni, L. baculus, L. cerradicula, L.
complexa, L. sallesi e L. saulensis. For peridomicile were collected
32 species totaling 3,630 sand flies (87.8%), in house indoor were
collected 13 species, totaling 343 sand flies (8.2%), outside the home
14 species, totaling 141 sand flies (3.4%) and in the wild area 7
species, totaling 27 sand flies (0.6%). The predominant species
caught from these areas was L. (L.) longipalpis, with 57% of the total
species collected inside the home, 45.2 % and 50.3 % peridomiciles
outside the home. The exception was the wild area with 29.5% of L.
(L.) longipalpis, the specimens that match with other species
represents 70.4 %. It is suggested that the transmission cycle of AVL
in the urban area is apparently frequented by the sand fly vector
associated with the presence of autochthonous cases.
N° 26 (1) 2015
SAND FLY FAUNA (DIPTERA: PSYCHODIDAE) IN
THE ENDEMIC AREA OF LEISHMANIASIS IN BARRA
D O G A R Ç A S – M ATO G R O S S O A N D T H E
DEMOGRAPHIC INFORMATION ON LUTZOMYIA
(LUTZOMYIA) LONGIPALPIS AND LUTZOMYIA
(NYSSOMYIA) WHITMANI
THE ABUNDANCE AND
DISTRIBUTION OF
PHLEBOTOMINE SAND FLY IN SICILY
Torina A.1,2; Reale S.1, 2; Blanda M.1; La Russa F.1; Intile S.1; Lelli R.1;
Vitale F.1
1
Istituto Zooprofilattico Sperimentale della Sicilia, Palermo, Italy;
University of Messina, Italy.
Sinara Cristina de Moraes, Lucas Heber Mariano dos Santos, Thiago
Nunes Alexandre, Thais Brito do Nascimento
100
2
Vet. Sc. Dep.
stefano.reale@izssicilia.it
Depto. de Parasitologia/ICB/UFMG, Belo Horizonte- MG
sinaramoraes@hotmail.com
Keywords: Leishmaniasis, sandflies, Leishmania
Sand flies are transmitting a number of diseases, including
leishmaniasis. The objective of the study was to assess the sand fly
fauna present in the neighbourhoods of Barra do Garças - MT and its
associate richness and population density with demographic data; to
determine the neighbourhoods with the highest risk of transmission of
leishmaniasis. Sand flies were captured from 2013 using CDC light
traps. The light traps were used in peridomestic areas near animal
shelters for three consecutive nights from 06:00 pm-06:00 am of 41
neighbourhoods. The captured specimens were fixed in 70% alcohol
and identified in accordance with Young & Duncan. Demographic
data were collected from localities system - SISLOC. The diverse
phlebotomine fauna identified from 1650 specimens include 31
species under 2 genera, Shannon diversity H = 2.619, these were the
most abundant species Lutzomyia (Nyssomyia) whitmani (48.42%) ,
followed by L.(Lutzomyia) longipalpis (16.84%) , L.(L.) cruzi
(10.9%), L. lenti (6.96%) and a further twenty seven (<3%) as B.
pintoi, Brumptomyia brumpti, L. abunaensis , L. aconthopharynx, L.
aragaoi L. brasiliensis, L. carmelinoi, L. carrerai carrerai, L.
cerradincula, L. christenseni, L. davisi, L. evandroi, L. ferreirana, L.
flaviscutellata, L. goiana, L. hermanlenti , L. longipennis, L. lutziana,
L. oliveirai, L. punctigeniculata, L. runoides, L. sallesi, L. saulensis,
L. sordellii , L. teratodes, L. termitophila e L. walkeri. The
neighbourhoods with the highest densities of the species L. (L.)
longipalpis and L. (N.) whitmani were: DERMAT with 9.3% and
57.35%, São José with 37.80% and 13.62%, Palmares with 22.15%
and 7.16%, Vila Maria 11.26% and 0.35% and Novo Horizonte with
2.63% and 10%; corresponding to 14.5% inhabitants. The urban area
have plots free and some neighbourhoods make border with
conservation area Serra Azul Park State. It's common the presence of
chickens. The sandfly fauna in Barra do Garças is varied and some
species identified are considered to be potentials vectors of American
cutaneous leishmaniasis (ACL) in Brazil, as L. flaviscutellata, L.
wellcomei and L. (N.) whitmani, or vectors of American visceral
leishmaniasis (AVL), as L. (L.) longipalpis, L. (L.) cruzi. The
neighbourhood's studied, 38 of them had at least one related species
transmission of leishmaniasis. The data suggest that the cycle of
leishmaniasis in the region is highly urbanized, as the wide
distribution of vectors in the urban area.
Keywords: Epidemiology, Phlebotomus, Sergentomya
Sand flies (Psychodidae: Phlebotominae), the vectors of Leishmania
as well as of phleboviruses, have been known in Mediterranean for
long time. In this paper the diversity and the abundance of
phlebotomine vectors were investigated in Sicily through the 2012
sand fly season using Onderstepoort-type backlight traps. Traps were
set up from 1 h before sunset to 1 h after sunrise and collected the next
morning. The largest number of sand flies captured in one night was
11396 in the capture of 9th, July, while the lowest number was 1 in
catches of April and December. June and July were the month with the
highest abundance (n.of insect/n. of positive catches), in the other
months there was a progressive decrease. P. perniciosus only had a
trend characterized by two peaks, one in June and the second slight
increase in August, P. mascittii had a peak in June, while the other
species [what are they] showed a peak in July. A total of 101123 sand
flies was identified as belonging to 7 species: Phlebotomus
perfiliewi (40.31%), P. perniciosus (24.84%), P. neglectus (16.67%),
P. mascittii (3.31%), P. papatasi (0.20%), P. ariasi (0,05 %) and
Sergentomyia minuta (14.62%). The two species considered as the
main vectors of human leishmaniasis in Italy, P. perniciosus and P.
perfiliewi, were the most abundant, although the higher prevalence
of P. perfiliwi can be explained by the fact that the catch was made in
rural areas and P. perniciosus is the main anthropophilic vector, while
P. perfiliewi is considered mainly zoophilic. Moreover, P. neglectus is
a proven vector of L. infantum in Greece. In conclusion the risk of
transmission of Leishmania in Sicily remains high and competent
vectors are well represented and it can be seen a prolonged season of
sand flies and an increase of the population in terms of diversity of
identified species, abundance and prevalence. Acknowledgements We thank Rosa Filippi for the helpful and precious technical support
and Gabriella Serafino Agrusa.
N° 26 (1) 2015
LUTZOMYIA LONGIPALPIS DISTRIBUTION AND
ABUNDANCE IN THE CITY OF CORRIENTES,
NORTHEASTERN ARGENTINA
A STUDY OF PHLEBOTOMINAE IN MODIFIED
WHITE AND BLACK SHANNON TRAPS IN A
MAROON AREA AT PIRAPUTANGA DISTRICT,
AQUIDAUNA, MATO GROSSO DO SUL, BRAZIL
Berrozpe Pablo, Araujo Analía, Santini Maria Soledad, Utges Maria
Eugenia; Salomon Oscar Daniel
REDILA- Instituto Nacional de Medicina Tropical; Centro Nacional de Diagnostico e
Investigación en Endemoepidemias; CONICET
peberrozpe@gmail.com
Keywords: Lu. longipalpis, environmental stratification.
Corrientes was sampled in two stages during summer. In the first stage
(December 2013), the city was divided in areas of 800 x 800 m to
evaluate Lutzomyia longipalpis distribution in the whole city. In the
second stage (February-March 2014) we evaluated the distribution of
Lu.longipalpis abundance according to environmental strata. A total
of 6 strata were discriminated based on satellite images 'Spot 5' HRG1.
In each stratum we selected at least three positive sites for Lu.
longipalpis (first stage samples). In each peridomestic environment
selected under 'worst case' criteria, mini-traps of UV-light were
located during 5 consecutive nights from 5 p.m. to 9a.m. (first stage:
51 traps/night; second stage: 21 traps/night. At each peridomicile we
recorded maximum and minimum temperature and relative humidity,
and the georeference. The specimens were clarified with lacto-phenol
and identified according to Galati (2003). A total of 1390 sandflies
were captured (555 in the first stage, and 835 in the second stage). In
the first stage a total of 5 species were captured: Lu. longipalpis
(58.25%), Nyssomyia neivai (31.75%), Mygonemyia migonei
(7.40%), Evandromyia cortelezzii complex (1.0%) and
Mycropigomyia quinquefer (1.6%). In the second stage 4 species were
captured: Lu. longipalpis (94.3%), Ny. neivai (2.7%), Mg.
migonei(2.3%), Ev. cortelezzii complex (0.6%). These preliminary
results showed that Lu. longipalpis is the most abundant specie in the
city of Corrientes. When we analyzed the environmental strata, we
could describe a decreasing gradient of abundance from primary
vegetation towards the most densely inhabited center of the city. We
also found areas of high (>30 Lu. longipalpis individuals/trap/night),
medium (15 to 30 Lu. longipalpis individuals/trap/night) and low
abundance (< 15 Lu. longipalpis individuals/trap/night) in sites
associated to primary riparian vegetation of the River Paraná. Areas
whithin medium and low abundance of Lu.longipalpis in peripheral
neighborhoods and finally areas of low abundance of Lu. longipalpis
in Downtown.
101
Jucelei de Oliveira Moura Infran1,4; Daiana Alovisi Souza1; Wagner
Fernandes de Souza1; Aline Etelvina Casaril2; Gabriel Utida Eguchi3;
Elisa Teruya Oshiro4; Carlos Eurico Fernandes1; Alessandra Gutierrez
de Oliveira1,2,4
1
.Curso de Pós-graduação em Biologia Animal - Centro de Ciências Biológicas e da
Saúde/Universidade Federal de Mato Grosso do Sul, 2.Curso de Pós-graduação em
Doenças Infecciosas e Parasitárias- Faculdade de Medicina/ Universidade Federal
de Mato Grosso do Sul; 3.Faculdade de Medicina Veterinária e Zootecnia Universidade Federal de Mato Grosso do Sul; 4.Laboratório de Parasitologia-Centro
de Ciências Biológicas e da Saúde/Universidade Federal de Mato Grosso do Sul
Cidade Universitária, 79070-900, Campo Grande, MS, Brazil.
alessandra.oliveira@ufms.br
Keywords: sand flies, Shannon trap, anthropophily
The study was carried out at Furnas dos Baianos, a Maroon area
located at the district of Piraputanga in Aquidauana, Mato Grosso do
Sul-MS, Brazil. From September 2013 to June 2014, uninterrupted
captures were performed for 24h on a monthly basis in modified
white and black Shannon traps placed at the foot of the mountain.
Temperature and relative humidity were recorded during this period.
A total of 707 sand flies was captured, 269 (39.03% males and
60.96% females) in white Shannon trap and 438 (62.32% males and
37. 67% females) in black shannon trap. 11 species were identified as:
Brumptomyia avellari, Evandromyia lenti, Evandromyia saulensis,
Lutzomyia díspar, Lu. longipalpis, Martinsmyia oliveirai,
Micropygomyia peresi, Nyssomyia whitmani, Pintomyia misionensis,
Psathyromyia bigeniculata. Lu. longipalpis was the most frequently
found in both colour traps, followed by Ny. whitmani. The third most
prevalent species was Pi. misionensis in the white Shannon trap and
Lu. dispar in the black Shannon trap. Along the captures the
researches realized they were attracting many sand flies and they
were identified as Lu. longipalpis, Lu. dispar and Pi. misionensis.
Interestingly many males (80.85%) of Lu. longipalpis were among
them.
N° 26 (1) 2015
NYCTHEMERAL RHYTHM OF PHLEBOTOMINAE IN
A MAROON AREA AT PIRAPUTANGA DISTRICT,
AQUIDAUNA, MATO GROSSO DO SUL, BRAZIL
SAND FLY FAUNA IN CHAPARE, BOLIVIA: AN
ENDEMIC FOCUS OF LEISHMANIA (VIANNIA)
BRAZILIENSIS
Jucelei de Oliveira Moura Infran1,4; Daiana Alovisi Souza1; Wagner
Fernandes de Souza1; Aline Etelvina Casaril2; Gabriel Utida Eguchi3;
Elisa Teruya Oshiro4; Carlos Eurico Fernandes1; Alessandra Gutierrez
de Oliveira 1,2,4
Marinely Bustamante Gomez1, Mery Diaz1, Jorge Espinoza1, Rudy
Parrado1, Richard Reithinger2, 3, and Ana Lineth Garciá1
1
. Curso de Pós-graduação em Biologia Animal - Centro de Ciências Biológicas e da
Saúde/Universidade Federal de Mato Grosso do Sul, 2. Curso de Pós-graduação em
Doenças Infecciosas e Parasitárias- Faculdade de Medicina/ Universidade Federal de
Mato Grosso do Sul; 3. Faculdade de Medicina Veterinária e Zootecnia - Universidade
Federal de Mato Grosso do Sul; 4. Laboratório de Parasitologia-Centro de Ciências
Biológicas e da Saúde/Universidade Federal de Mato Grosso do Sul Cidade
Universitária, 79070-900, Campo Grande, MS, Brazil.
102
1
IIBISMED, Facultad de Medicina, Universidad Mayor de San Simón, Cochabamba,
Bolivia; 2Faculty of Infectious & Tropical Diseases, London School of Hygiene &
Tropical Medicine, Keppel Street, London WC1E 7HT, United Kingdom; 3George
Washington University School of Medicine and Health Sciences, Washington, D.C.
20035.
marinelybustamante@gmail.com
Keywords: leishmaniasis, Lutzomyia, Leishmania (Viannia), sand
fly
Keywords: sand flies, hourly activity, seasonality
The study was carried out at Furnas dos Baianos, a Maroon area
located at the district of Piraputanga, in Aquidauana, Mato Grosso do
Sul (MS), Brazil. From July 2012 to June 2014, uninterrupted captures
in modified Shannon trap placed at the foot of the mountain were
performed for 24h on a monthly basis for two years. Temperature and
relative humidity were recorded during this period. A total of 1.815
sand flies were captured and the following species were identified:
Nyssomyia whitmani, Lutzomyia dispar, Lutzomyia longipalpis,
Pintomyia misionensis, Psathyromyia bigeniculata, Evandromyia
lenti, Micropygomyia oswaldoi, Brumptomyia avellari,
Micropygomyia peresi, Martinsmyia oliveirai, Evandromyia
saulensis, Pintomyia pessoai, Psychodopygus davisi, Evandromyia
evandroi and Psathyromyia campograndensis. Nyssomyia whitmani
(28.4%, 515 ∕ 1,815, with 51.0% of females and 49.0% of males) and
Lu. longipalpis (20.4%, 371 ∕ 1,815, with 25.6 % of females and
74.4% of males) were the first and third most prevalent species, also
they showed highest abundance of insects captured in dry season,
being the activity time between 6pm - 5am and 7pm - 5am,
respectively. These two species deserve attention because they are
considered main vectors of Leishmania (Viannia) braziliensis and
Leishmania (Leishmania) infantum in many regions of Brazil
including Mato Grosso do Sul. Lu. dispar, (24.7%, 455 ∕ 1,815, with
98.5% of females and 1.5% of males), the second most prevalent
species was found high activity in rainy season, between 6pm and
2am.
Data on the distribution and abundance of Lutzomyia spp. (Diptera:
Psychodidae) in Bolivia is scarce. Sand flies from an area of
Leishmania (Viannia) braziliensis endemicity in the Isiboro-Secure
National Park in the Department of Cochabamba were captured and
identified to species. In total, 945 sand flies (789 females and 156
males) belonging to 15 species were collected from the four
collection points in two study villages in 2007. With 549 (58.1%)
specimens, Lutzomyia shawi was the most abundant species,
followed by Lutzomyia (Trichophoromyia) sp. (22.2%), Lutzomyia
llanosmartinsi (8.3%), Lutzomyia antunesi (4.3%), and Lutzomyia
olmeca (2.1%). Abundance and species composition varied between
rainy and dry seasons, with 99.3% of all sand flies being collected
outdoors. Because of species abundance and confirmed Leishmania
infection in previous entomological collections, we believe Lu. shawi
is the vector of L. (Viannia) braziliensis in Isiboro-Secure National
Park.
N° 26 (1) 2015
DIVERSITY AND NYCTHEMERAL RHYTHM OF
PHLEBOTOMINES IN SAN JULIAN COMMUNITY
(COCHABAMBA, BOLIVIA)
STUDIES ON THE FAUNA OF SAND FLIES (DIPTERA:
PSYCHODIDAE: PHLEBOTOMINAE) IN CORUMBÁ,
MATO GROSSO DO SUL, BRAZIL
Jorge Espinoza Echeverria; Marinely Bustamante; Mery Diaz; Rudy
Parrado; Ana Lineth Garciá
Aline Etelvina Casaril1; Everton Falcão de Oliveira2; Neiva Zandonaide
Monaco3; Gabriel Utida Eguchi4; Helen Rezende de Figueiredo1;
Nathália Lopes Fontoura Mateus4; Wagner de Souza Fernandes1;
Suellem Petilim Gomes1; Daiana Alovisi Souza5; Jucelei de Oliveira
Moura Infran5,6; Elisa Teruya Oshiro6; Eunice Aparecida Bianchi
Galati2; Alessandra Gutierrez de Oliveira1,5,6
IIBISMED, Facultad de Medicina, Universidad Mayor de San Simón, Cochabamba,
Bolivia
jorgeespinozaecheverria@gmail.com
Keywords: Leishmaniasis, Sand flies, Nycterohemeral,
Peridomestic.
The dynamics of different forms of the disease leishmaniasis is
correlated with fluctuation of the populations of hosts and vectors,
such as changes in the environment and climatic factors such as
temperature, humidity and rainfall. Usually generated by the
transformations of the wild environment through deforestation and
replacement of wild animals by synanthropic, which facilitates the
adaptation of some vector species colonized the environment and
promoting an expansion of leishmaniasis transmission area of the
forest to the home environment. Sand flies were captured and
identified to species, from an area of Leishmania (Viannia)
braziliensis endemicity in the Isiboro-Secure National Park in the
Department of Cochabamba, during the season dry and humid, in
peridomestic environments. Were determined the diversity of species
during the nycthemeral rhytm of sand flies. The statistical analyzes
beyond the ecological indices will be analyzed with the help of R
package and Sigmaplot. Were captured 236 individuals belonging to 7
species of the genus Lutzomyia. Lu. shawi was the most abundant
species during the two seasons with 52.1% in the dry season and
63.7% in the humid season. Was observed higher nycthemeral rhytm
among the 20 and 21 hours; the activity of sand flies, decrease as a
function to which the temperature decreases and relative humidity
increases. During the humid season, present correlation positive
among temperature and abundance (r2=0,744), but correlation
negative among humidity and abundance (r2= 0,692). The diversity of
species Shannon–Weaver, in the tow season showed, was of the
H=0,603043 and equability of Pielou was the J= 0,87000.
103
1
Curso de Pós-Graduação em Doenças Infecciosas e Parasitárias - Faculdade de
Medicina/ Universidade Federal de Mato Grosso do Sul; 2Departamento de
Epidemiologia, Escola de Saúde Pública, Universidade de São Paulo; 3Centro de
Controle de Zoonoses, Secretaria de Saúde do Município de Corumbá; 4Faculdade de
Medicina Veterinária e Zootecnia - Universidade Federal de Mato Grosso do Sul;
5
Curso de Pós-Graduação em Biologia Animal - Centro de Ciências Biológicas e da
Saúde/Universidade Federal de Mato Grosso do Sul; 6Laboratório de ParasitologiaCentro de Ciências Biológicas e da Saúde/Universidade Federal de Mato Grosso do
Sul Cidade Universitária, 79070-900, Campo Grande, MS, Brasil.
Keywords: sand flies, abundance, diversity
The municipality of Corumbá is considered area of intensive
transmission for Visceral Leishmaniasis (VL) and reported 200 cases
of the disease during 1999- 2011 and seven confirmed cases of
American Cutaneous Leishmaniasis (ACL) from 2007 to 2011.
Fauna studies leading up to the association between the occurrence of
these insects and the prevalence of the leishmaniasis in the areas with
records of the diseases and promoting measures for the control and
surveillance. The aim of this study was to identify the sand flies
(Diptera: Psychodidae: Phlebotominae) fauna in the Municipality of
Corumbá, Mato Grosso do Sul, Brazil. The Sand flies were collected
weekly with light automatic traps at 06:00 pm to 07:00 am, in peri and
intradomicile environments in five urban districts, from April 2012 to
March 2013. The sand flies were sent to the Laboratory of
Parasitology at the Federal University of Mato Grosso do Sul. They
were clarified and identified following the classification proposed by
Galati (2003). The 390 weekly captures yielded 7,370 specimens of
phlebotomine, 6,169 males (83.70%) and 1,201 females (16.30%).
The fauna was composed by 12 species including: Brumptomyia
brumpti, Evandromyia aldafalcaoae, Evandromyia cortelezzii,
Evandromyia corumbaensis, Evandromyia sallesi, Evandromyia
walkeri, Lutzomyia cruzi, Lutzomyia forattinii, Micropygomyia
peresi, Martinsmyia oliveirai, Psathyromyia bigeniculata and
Sciopemyia sordellii. The most abundant species was Lu. cruzi
representing 93.79% and the second was Lu. forattinii with 3.53%.
Overall, the insects were more frequently captured in peridomestic
(60.45%) than indoors (39.55%) of the residences.
N° 26 (1) 2015
ENTOMOLOGICAL SURVEILLANCE OF
PHLEBOTOMINE SAND FLIES VECTORS OF
LEISHMANIASIS IN AREAS IMPACTED BY THE
TRANSFER OF THE SÃO FRANCISCO RIVER
PROJECT, IN THE STATE OF CEARÁ, BRAZIL
THE INFLUENCE OF MOON LIGHT AND LUNAR
PERIODICITY ON THE DENSITY OF PHLEBOTOMUS
(LARROUSSIUS) ORIENTALIS PARROT, 1936
(DIPTERA: PSYCHODIDAE) FROM LIGHT TRAP
COLLECTIONS IN ETHIOPIA
Júlia dos Santos Silva1, Antonio Pereira dos Santos2, Fabrício Kássio
Moura Santos2, Luiz Osvaldo Rodrigues da Silva2, Lindemberg
Caranha3, Elizabeth Ferreira Rangel1
Araya Gebresilassie1, Oscar David Kirstein2, Aviad Moncaz2, Habte
Tekie1, Meshesha Balkew3, Alon Warburg2, Asrat Hailu4, and Teshome
Gebre-Michael3
1
1
Department of Zoological Sciences, Addis Ababa University, Addis Ababa, Ethiopia,
Ethiopia; 2Department of Microbiology and Molecular Genetics, The Institute of
Medical Research Israel-Canada The Kuvin Center for the Study of Infectious and
Tropical Diseases, Faculty of Medicine, The Hebrew University, Hadassah Medical
School, Jerusalem, Israel; 3Aklilu Lemma Institute of Pathobiology, Addis Ababa
University, Addis Ababa, Ethiopia; 4Department of Microbiology, Immunology and
Parasitology, Faculty of Medicine, Addis Ababa University, Addis Ababa, Ethiopia
Laboratório de Transmissores de Leishmanioses, Instituto Oswaldo Cruz, FIOCRUZ,
Av. Brasil, 4365, 21040-360, Manguinhos, Rio de Janeiro, Brazil; 2Secretaria de Saúde
do Estado do Ceará, Brazil; 3Laboratório de Vetores, Reservatórios e Animais
Peçonhentos Dr. Thomaz Correa Aragão, SESA/CE, Brazil.
juliass@ioc.fiocruz.br
Keywords: Phlebotominae, species diversity, environmental
impact, vector ecology
104
shambe2006@yahoo.com
Keywords: Density, light traps, lunar cycles, P. orientalis
American Cutaneous Leishmaniasis (ACL) and American Visceral
Leishmaniasis (AVL) are relevant public health problems in Brazil. In
addition to biological and physio-geographical factors, the
environmental impact resulting from human actions, such as
deforestation, construction of hydro-electric plants, migration
processes, among others, play important roles in transmission of
leishmaniasis. In Brazilian north-eastern state of Ceará, both ACL and
AVL are endemic diseases, the latter presenting intense transmission.
The Transfer of the São Francisco River project (TSFR) aims to
transform the socio-economic context of the Northeast. However,
some negative impacts have been discussed. From the point of view of
public health, there is a hypothesis of epidemic outbreaks of ACL and
AVL. The purpose of this study was the entomologic surveillance of
Phlebotominae, to subsidise prevention and control actions for
leishmaniasis in towns influenced by the TSFR in state of Ceará.
Monitoring sites were selected in the towns of Jaguaretama,
Jaguaribe, Lavras da Mangabeira, Brejo Santo, Mauriti, Iguatu and
MissãoVelha. Sand flies were captured monthly from May 2011 to
December 2013, with three HP light traps in each site, installed in and
around household and in the woods, working for 12 hours from sunset
for three consecutive nights. Data analyses used were indexes of
richness, dominance, abundance and diversity of species. Taking into
consideration the overall captures and analysing intra- and
peridomiciliary areas and woods separately, Lutzomyia longipalpis
(Lutz & Neiva, 1912), the main vector of AVL, was the dominant and
most abundant species in all months, in all towns. In order, the other
most frequent species were Lutzomyia lenti (Mangabeira, 1938),
Lutzomyia evandroi (Costa Lima &Antunes, 1936) and Lutzomyia
villelai (Mangabeira, 1942). Iguatu, a town with intense transmission
of AVL, presented the highest number of specimens, of which 96.2%
were Lu. longipalpis. The largest species richness was at Mauriti
(S=13), while the greatest diversity was found in MissãoVelha
(H´=1.26), towns with moderate transmission of AVL and ACL cases,
registering Lutzomyia intermedia (Lutz & Neiva, 1912), Lutzomyia
migonei (França, 1920) and Lutzomyia whitmani (Antunes &
Coutinho, 1939), important vectors of ACL. Surveillance and control
activities for AVL and ACL and education actions in the involved
towns are advised to reduce the transmission risk. Financial support:
FIOCRUZ
Various environmental variables including moon light and lunar
periodicity have been shown to affect the night activity of
phototrophic dipteran flies. To investigate the possible effect of lunar
phases on the activity of phlebotomine sand flies, trapping of sand fly
species was conducted during December 2012 to June 2013 in Tahtay
Adiyabo District, Northern Ethiopia. Sand flies were collected from
peri-domestic and agricultural fields using CDC miniature light traps
and sticky traps on 28 nights, divided among each of the 4 lunar
phases. A total of 13,460 sand flies (11,633 in light traps and 1,827 in
sticky traps) belonging to eight species of the genus Phlebotomus (P.
orientalis, P. bergeroti, P. rodhaini, P. duboscqi, P. papatasi, P.
martini, P. lesleyae and P. heischi) were captured. The predominant
species in both trapping sites and in all lunar phases was P. orientalis,
constituting 98.3% of the specimens. A statistically significant
difference was observed in the density of P. orientalis collected by
CDC light traps among lunar phases (P<0.05). The highest mean
number (231.13±36.27 per trap/night) of P. orientalis was captured
during the new moon phases, in the absence of moonlight.
Significantly, fewer sand flies were attracted in light traps during full
moon. However, P. orientalis and the other sand fly species from
sticky traps did not differ in their density among the four lunar phases
(P=0.122). The blood feeding of P. orientalis was not influenced by
the presence or absence of moon light. Results of the current study
showed that the attraction and trapping efficiency of CDC miniature
light traps is substantially influenced by moon illumination.
Accordingly, sampling of sand fly species using light traps to estimate
their population density in the field should take the effect of moon
light and lunar periodicity into account on the trapping efficacy of
light traps.
N° 26 (1) 2015
ID 92-P Vector Competence & Infections
CHARACTERISTICS OF HUMAN DWELLINGS AND
THEIR SURROUNDINGS THAT FAVOUR THE
ABUNDANCE OF PHLEBOTOMINES IN FOUR URBAN
AREAS OF THE CARIBBEAN COAST OF COLOMBIA
PHLEBOTOMINE SAND FLY FAUNA (DIPTERA:
PSYCHODIDAE) AND NATURAL FLAGELLATE
INFECTIONS FROM AN AREA HIGHLY ENDEMIC
FOR CUTANEOUS LEISHMANIASIS ON THE
BRAZILIAN-BOLIVIAN FRONTIER IN THE STATE
OF ACRE, BRAZIL
Jorge Rodríguez-Jiménez1, Alveiro Pérez-Doria1, Luis Romero-Ricardo1,
José Vergara1, Ana Montesino1, Lily Martínez1, Luis Cortés2, Suljey
Cochero3 & Eduar E. Bejarano1
1
105
Iorlando da Rocha Barata, Maria Sueli Barros Pinheiro, Fábio Márcio
Medeiros da Silva, Roberto Carlos Feitosa Brandão, Thiago Vasconcelos
dos Santos, Fernando Tobias Silveira
Grupo de Investigaciones Biomédicas, Universidad de Sucre, Sincelejo, Colombia.
2
Secretaría de Salud Departamental de Bolívar, Cartagena, Colombia. 3Secretaría de
Salud Departamental de Sucre, Sincelejo, Colombia
Instituto Evandro Chagas/ SVS/ MS, Ananindeua, Pa, Brazil
Keywords: Lutzomyia, cutaneous leishmaniasis, urban
transmission, Colombia
Keywords: phlebotomine, flagellate, Amazonian frontier, cutaneous
leishmaniasis
The transmission of Leishmania is no longer exclusive to sylvatic and
rural habitats, with new epidemiological scenarios arising in urban
areas as a consequence of natural environmental changes or those
brought about by human activities and favouring the survival or
adaptation of sand fly vectors of the parasite. In the present study the
characteristics of human dwellings and the peridomiciliary
environment were analyzed in relation to the abundance of
phlebotomines of the genus Lutzomyia in four urban areas of the
Caribbean coast of Colombia, from which cases of cutaneous and
visceral leishmaniasis had recently been reported. This involved the
selection of 80 dwellings in the municipalities of Sincelejo, Ovejas, El
Carmen de Bolívar and Cartagena. Visits were made to houses in
which the characteristics of each dwelling and its surroundings were
recorded. In addition insects were collected using CDC light traps
hung inside and in the peridomiciliary area of each house from 18:00
to 06:00 on two consecutive nights. In all 9042 specimens of the genus
Lutzomyia were captured belonging to 11 species, of which Lutzomyia
evansi represented 93.3% of the total and was by far the most abundant
in the four municipalities. The remaining phlebotomine fauna was
represented by Lu. atroclavata (0.4%), Lu. carpenteri (0.04), Lu.
cayennensis cayennensis (1.5%), Lu. dubitans (1.4%), Lu. gomezi
(2.3%), Lu. panamensis (2.8%), Lu. rangeliana (0.2%), Lu.
trinidadensis (0.4%), Lu. venezuelensis (0.4%) and Lu. micropyga
(0.04%). Multiple correspondence analysis was used to determine that
the following characteristics favoured the high abundance of
phlebotomines in and around houses in these urban centres: (1)
proximity (0-10 m) of dwellings to forested areas; (2) presence of
bushes, grass and leaf litter in patios; (3) walls made of “bahareque”
(adobe) and dirt floors; (4) accumulation of solid residues in open
areas; (5) disposal of human waste in the open air; and (6) having
electrical power in the dwellings. As measures to reduce
phlebotomine density and the risk of urban Leishmania transmission,
sanitary conditions in the study areas should be improved to eliminate
sources of organic material in which immature sand flies develop. The
construction of dwellings should also be modified to include other
materials.
Knowledge of the phlebotomine sand fly fauna in foci of cutaneous
leishmaniasis (CL) is essential for epidemiological surveillance,
providing information on the main existing species and their
distribution. The increasing number of cases of CL in Amazonia, due
mainly to environmental changes and high migratory flows, leads to
the prioritization of surveillance at strategic areas, such as the
Amazonian frontiers. Thus, the aim of this study was to identify the
phlebotomine sand fly fauna and find natural flagellate infections in
an area highly endemic for CL at the Brazilian-Bolivian frontier. The
study was carried out in a rural area, namely Brasiléia municipality
(S11°00'36” W68º44'52”), state of Acre (AC), Amazonian Brazil.
During 2012 and 2013, four collections (15 days each) were
undertaken in the dry and rainy seasons with eight CDC light traps
disposed at ground level (1.5 meters) and two in the forest canopy
(20m) from 18:00 to06:00 hours. Females were dissected under
sterile conditions and each homogenate of infected midgut was twofold inoculated into Difco B45 media. Both sexes were identified
according to taxonomic criteria (Young and Duncan, 1994). A total of
3,625 phlebotomine sand flies belonging to 41 species were
collected. Females (n= 2,793) predominated over males (n= 1,026).
The collections were more abundant in the rainy season (n= 2,716)
than in the dry season (n = 1,103). Ten species were noted as being
potentially implicated in the transmission of CL. Among them, the
five most frequent were Lutzomyia whitmani (n = 878; 24.2%), Lu.
auraensis (n= 767/ 21,1%), Lu. ubiquitalis (n= 289/ 7,9%), Lu. davisi
(213/ 5,8%) and Lu. ntunesi (163;/ 4,4%). A single female of Lu.
whithmani and two of Lu. yuilli yuilli were found harboring
flagellates with a overall infection rate of 0,1%. No parasite isolation
was accomplished successfully. In conclusion, this study leads to
confirm the potential implication of Lu. whitmani and Lu. yuilli yuilli
in the epidemiology of CL at AC and in addition, despite no natural
infection was found in Lu. auraensis, its high frequency also adds
weight to the hypothesis on the participation of this specie in the
transmission of the disease, as has been suggested for Amazonian
Peru.
N° 26 (1) 2015
ID 93-P Vector Competence & Infections
ID 94-P Vector competence & Infections
SANDFLIES (DIPTERA: PSYCHODIDAE)
NATURALLY INFECTED BY LEISHMANIA IN RURAL
AREAS OF THE CITY OF NOVA MUTUM, MATO
GROSSO, BRASIL
SAND FLY FAUNA (DIPTERA, PSYCHODIDAE)
COLLECTED IN THE MIDDLE PURUS RIVER
REGION, AMAZONAS, BRAZIL
Sirlei Franck Thies1,2,3, Ana Lucia Maria Ribeiro3,5, Jorge Senatore
Rodrigues Vargas3,5, Rosina Djunko Miyazaki4, Érika Monteiro
Michalsky6, Edelberto Santos Dias6
1
UFMT, Cuiabá/MT. 4Laboratory of Leishmaniasis, Center of Research René
Rachou/Fiocruz-Minas Gerais. 5University Hospital Júlio Müller, UFMT, Cuiabá/MT.
106
Túllio Romão R. da Silva, Mauro Diego Gobira Guimarães de Assis,
Felipe Dutra Rêgo, Maíra Posteraro Freire, Paloma Helena Fernandes
Shimabukuro
Centro de Pesquisas René Rachou/FIOCRUZ MINAS
palomahfs@gmail.com
Keywords: Phlebotominae, Fauna, Amazonas State
sfthies@hotmail.com
Keywords: Leishmania, Lutzomyia, Sandflies.
The leishmaniases are zoonotic diseases caused by protozoa of the
Kinetoplastida order, Trypanosomatidae family, Leishmania genus
that affect humans, and are transmitted in the Americas by insects
belonging to the Lutzomyia genus, called sandflies. The present study
aimed to identify the sandfly fauna and to verify their natural infection
by Leishmania species in a rural area in the city of Nova Mutum, Mato
Grosso, a place with notification of human cases of American
Cutaneous Leishmaniasis (ACL). Entomological collections were
made every two months, from June 2011 to April 2012, with CDC
light traps which were installed at 6 p.m. and collected at 7 a.m. the
next day, in ten specific places 100 meters distant from each other,
from the edge to the inside of the forest. The captured insects were
sorted, dissected, prepared and identified according to Young and
Duncan (1994). Sandfly pools contained between 1 and 10 female
insects, separated by species, trap number, place and date of capture.
DNA extraction was performed with Extraction Kit of Tissues and
Cells from GE Amersham Biosciences. Positive PCR bands of
Leishmania species were purified using the commercial kit Qiagen.
Genetic sequencing was performed via ABI 3130 platform. The
Standard Nucleotide Blast program was used for analysis of
bioinformatics and the sequences were compared to GenBank. The
presence of 3,743 specimens belonging to 30 different species was
found. The most abundant was Lutzomyia antunesi (45.44%), which
was found in all months and capture places. Females constituted
73.15% of the specimens. The highest densities occurred in October
2011, during the rainy season in the state. Two hundred and ninetythree (293) pools of females were assembled, belonging to six
different species, and L. antunesi was predominant (196/293).
Thirteen (13) pools showed natural infection, corresponding to a
minimum rate of infection of 1.01%. Lutzomyia ubiquitalis presented
two pools infected either with Leishmania braziliensis or Leishmania
guyanensis. Lutzomyia antunesi presented six pools infected either
with Le. braziliensis or Le . guyanensis, four pools did not allow the
alignment of the sequences (Leishmania species) and one pool
infected with Leishmania infantum. These infected sandfly species
are suspected vectors in the transmission cycle of ACL in this location,
where the presence of Le. infantum poses the risk of transmission of
Visceral Leishmaniasis.
Phlebotomine sand flies are of medical importance because they are
vectors of human pathogens, such as protozoa of the genus
Leishmania Ross, etiological agent of American cutaneous
leishmaniasis (ACL). In Lábrea, a municipality in the state of
Amazonas, Brazil, ACL is primarily associated with subsistence
activities, such as collection and extraction of forest products,
undertaken by both indigenous and non-indigenous people. Data on
ACL in indigenous populations are scarce, such that there is little
information on the identity of the etiologic agent(s), reservoir host(s)
and insect vector(s). The aim of this work was to study the sand fly
fauna collected in the Indigenous Reserve Caititu, Lábrea. A total of
1,267 sand flies were collected, of which 819 (64.6 %) were females
and 448 (35.4 %) males, from 10 genera and 32 species. The most
abundant genera were Psychodopygus (34.3%), Trichophoromyia
(22.9%)and Nyssomyia (15.3%). The most abundant species were
Trichophoromyia ubiquitalis (Mangabeira) (n = 235, 18.5%),
Psychodopygus davisi (Root) (n = 228, 18.0%) and Nyssomyia
antunesi (Coutinho) (n = 135, 10.7%). Direct sequencing of PCR
products demonstrated the presence of Leishmania (Leishmania)
amazonensis and Leishmania (Viannia) braziliensis in the following
species of sand flies: Evandromyia apurinan Shimabukuro, Silveira
& Silva, Ny. umbratilis (Ward & Fraiha), Ny. yuilli yuilli (Young &
Porter), Ps. davisi, Sciopemyia servulolimai (Damasceno & Causey)
and Th. ubiquitalis. The presence of natural infection by Leishmania
detected in the sand fly species investigated in this study suggests
their possible role in the transmission cycle of ACL in the study area.
N° 26 (1) 2015
MOLECULAR DETECTION OF LEISHMANIA IN
SAND FLIES (DIPTERA: PSYCHODIDAE) AND
S PAT I A L D I S T R I B U T I O N O F A M E R I C A N
CUTANEOUS LEISHMANIASIS IN THE XAKRIABÁ
INDIGENOUS RESERVE, BRAZIL
FIRST REPORT OF NATURAL PROMASTIGOTE
INFECTION OF SERGENTOMYIA DREYFUSSI
(DIPTERA: PSYCHODIDAE) SAND FLIES IN AN
ENDEMIC FOCUS OF CUTANEOUS LEISHMANIASIS
IN CENTRAL TUNISIA
Felipe Dutra Rego, Jerônimo Marteleto Nunes Rugani, Patrícia Flávia
Quaresma, Paloma Helena Fernandes Shimabukuro, Gabriel Barbosa
Tonelli, Ronaldo Guilherme Carvalho Scholte, Célia Maria Ferreira
Gontijo
Jomaa Chemkhi1, Rihab Yazidi2, Mahdi Driss3, Jihene Bettaieb2 and Afif
Ben Salah2
Grupo de Estudos em Leishmanioses. Centro de Pesquisas René Rachou, Fundação
Oswaldo Cruz. Belo Horizonte, Minas Gerais, Brasil.
107
1
Laboratoire d'Epidémiologie Moléculaire et Pathologie Expérimentale Appliquée
Aux Maladies Infectieuses, Institut Pasteur de Tunis, Université Tunis El Manar,
Tunisia. 2 Laboratoire d'Epidémiologie Médicale, Institut Pasteur de Tunis, Université
Tunis El Manar, Tunisia. 3 Plateforme de séquençage, Institut Pasteur de Tunis,
Université Tunis El Manar, Tunisia.
felipedutra@cpqrr.fiocruz.br
jomaa.chemkhi@pasteur.rns.tn
Keywords: Phlebotominae, Leishmania infection, Indigenous
Reserve, Kernel Density Estimator;
Autochthonous cases of American Cutaneous Leishmaniasis (ACL)
are reported since 2001 in Xakriabá Indigenous Reserve located at São
João das Missões, north of Minas Gerais state, characterizing this area
as endemic for this disease. This study aimed to investigate the
association between sand flies and the transmission of Leishmania
species. The data were correlated with epidemiological aspects
previously reported (natural infection in vertebrate hosts and human
cases of ACL) to identify hotspots related to infection by Leishmania
braziliensis, the most prevalent Leishmania species in the study area.
The sand flies were captured with HP light traps bimonthly for three
full days between October 2011 and August 2012 on trails previously
selected for the study of Leishmania reservoirs and sand fly fauna. The
DNA was extracted in pools, containing up to ten females from same
species and site colletion. The PCR was targeted to the internal
transcribed spacer 1 (ITS1) and amplicons were digested by the HaeIII
enzyme to analyze their restriction-fragment length polymorphisms
(RFLP). The spatial distribution of the ACL was analysed by Kernel
density to define the hotspots - concentration and pattern of ACL
infection - in the study area. A total of 4,031 female sand flies were
captured and grouped in 1,026 pools. Twelve pools were PCRpositive (1.16%) and Leishmania braziliensis infections were found in
Nyssomyia intermedia, Martinsmyia minasensis, Micropygomyia
capixaba and Micropygomyia peresi. Other Leishmania species
(Leishmania guyanensis, Leishmania infantum and Leishmania
amazonensis) were found in Evandromyia lenti, Mt. minasensis, Mi.
peresi and Ny. intermedia. The spatial analyses identify a main hotspot
indicating the location where the presence of Leishmania braziliensis
was more frequent. The results of natural infection in different hosts
studied by spatial analysis is an important tool to assess the areas with
high prevalence of parasite infection and may contribute to the choice
of specific preventive and control measures.
Keywords: Flagellate, infection, Sergentomyia, Tunisia
In order to study the natural infection of phlebotomine sandflies by
Leishmania parasites in central Tunisia where cutaneous
leishmaniasis is endemic, an entomological survey was carried out
during the transmission period from July to November 2013, in the
region of Mnara, belonging to the governorate of Kairouan. A total of
234 Phlebotomus papatasi, 89 P. longicuspis, 54 P. perniciosus, 44
Sergentomyia (antennata, 34 S. fallax, 28 S. dreyfussi and 12 S.
munita parroti were captured from Psammomys obesus burrows,
houses and animal shelters in a chenopod biotope, using CDC light
traps. Female sand flies were first dissected to look for natural
parasite infection in all parts of the gut. Then, they were
morphologically identified based on the keys of Lewis and Léger
(1983). Out of 230 females sand flies dissected, one identified as S.
dreyfussi and caught in an old house harboring various reptile lizards,
was found containing flagellates within the digestive tract. In vitro
isolation and culture of these promastigote forms allowed further
DNA extraction to be used for molecular analysis. PCR amplification
followed by restriction fragment length polymorphism (RFLP)
targeting ribosomal Internal Transcribed Spacer 1 (ITS1) DNA and
sequencing analyses indicated that the observed promastigotes are
probably reptile parasites belonging to the Trypanosomatidae family.
This study constitutes the first report in Tunisia on the natural
promastigote infection of Sergentomyia sand flies. This raises
questions about the precise promastigote species identification, and
more specifically to the possibility of the presence of
Sauroleishmania, suggested by the proximity to lizard habitats.
Further molecular analyses are ongoing to confirm this hypothesis.
This study received support from In Bio International Inc. grant N°:
W81XWH-09-C-0079.
N° 26 (1) 2015
SALIVARY GLAND HOMOGENATES FROM WILDCAUGHT LUTZOMYIA FLAVISCUTELLATA AND L.
(PSYCHODOPYGUS) COMPLEXUS DID NOT
EXACERBATE LEISHMANIA (L.) AMAZONENSIS AND
L. (V.) BRAZILIENSIS INFECTION IN BALB/C MICE
NATURAL INFECTION OF PHLEBOTOMINE SAND
FLIES (DIPTERA: PSYCHODIDAE) BY LEISHMANIA
ROSS (KINETOPLASTIDA: TRYPANOSOMATIDAE)
IN “TERRA FIRME” AND “VÁRZEA” ENVIROMENTS
IN TEFÉ MUNICIPALITY, AMAZONAS STATE,
BRAZIL
Laurenti MD, Francesquini FC, Passero LF, Tomokane TY, Carvalho
AK, Corbett CEP, Silveira FT
Medical School, University of São Paulo
mdlauren@usp.br
Keywords: Experimental Cutaneous Leishmaniasis, Pathology,
Lutzomyia flaviscutellata, Lutzomyia (Psychodopygus) complexus
108
Antonio Marque Pereira Júnior1,2; Carolina Bioni Garcia Teles3, Ana
Paula de Azevedo dos Santos3, Felipe Arley Costa Pessoa2, Jansen
Fernandes de Medeiros1,3
1
Inststuto nacional de Pesquisas da Amazônia, 2Centro de Pesquisa Leônidas & Maria
Deane, FIOCRUZ AMAZÔNIA, 3FIOCRUZ Noroeste.
facpessoa@amazonia.fiocruz.br
Keywords: Epidemiology, vectors, Leishmania lainsoni
During the natural transmission of leishmaniasis, the infected female
phlebotomine regurgitates promastigotes into the host's skin together
with the saliva. It has been reported that components of vector saliva
contain immunomodulatory properties that facilitate the
establishment of infection in the host, however most studies employed
salivary gland lysate (SGL) of laboratory colonized vectors. Thus, the
main objective of this study was to evaluate the specificity of SGL of
the phlebotomines Lutzomyia flaviscutellata and Lutzomyia
(Psychodopygus) complexus caught in the field for the infectivity of L.
(L.) amazonensis and L. (V.) braziliensis. BALB/c mice were
inoculated in the hind footpad with promastigotes of L. (L.)
amazonensis and L. (V.) braziliensis in the absence or presence of L.
flaviscutelata, and L. (P.) complexus SGL. The evolution of the lesion
size was evaluated weekly and biopsies from the site of infection were
collected for histopathological analysis and determination of parasite
load in the 4th and 8th week post infection (PI). Draining lymph nodes
were also collected to characterize subsets of T cells by flow
cytometry. L. (L.) amazonensis and L. (V.) braziliensis infections were
not exacerbated in the groups of mice co-inoculated with the SGL of
either phlebotomine species, in relation to the control group
inoculated only with the parasite. The lesion size and parasite burden
of the control group was higher or equal to the groups with saliva. In L.
(L.) amazonensis infections there was a decrease of CD4+ cells and an
increase in the population of CD8+ cells, while in L. (V.) braziliensis
infections there was a maintenance of CD4+ cells and an increase in
the population of CD8+ cells in all groups compared with the control
group. As a whole, the results show that the saliva of L. flaviscutellata
and P. complexus, in the natural vector/parasite binomium or not, did
not favor the establishment of the infection caused by L. (L.)
amazonensis e L. (V.) braziliensis in BALB/c mice. Supported by
CNPq, FAPESP and LIM50 HC-FMUSP.
Phlebotomine sandflies are insects of medical importance because
some species play a role in transmission of leishmaniasis. The aim of
this work was to detect natural infection and possible vectors in Tefé
municipality, Amazonas state, Brazil. Phlebotomine used were
collected using HP light traps during six months: January, February,
April, August, September and October, of 2013. To verify natural
infection, DNA samples were submitted to Polymerase chain reaction
(PCR) using kDNA gene and the identification of Leishmania species
were performed by PCR RFLP with hsp70 gene and genetic
sequence. A total of 1.679 was grouped in 60 samples of
Trichophoromyia ubiquitalis, 18 pools of Nyssomyia antunesi, one
pool of Ny. umbratilis, eight of Ny. yuilli yuilli, four of
Psychodopygus ayrozai and four of Ps. davisi. In kDNA region 14/95
samples were positive to presence Leishmania spp., ten sample
belonging to Th. ubiquitalis and four sample to Ps. davisi. The
minimal infection rate (MR) was 0.83%. In PCR to hsp70 gene was
observed seven sample positive of the 14 sample amplified for
kDNA. This sample belonging to Th. ubiquitalis. In PCR RFLP was
observed that samples submitted to Hae III enzyme show a pattern
between Leishmania (Viannia) lainsoni and Leishmania (V.) shawi.
The genetic analysis confirmed the specie L. (V.) lainsoni in Th.
ubiquitalis sample. The similarity of sequences was 99% compared to
reference strain deposited on GenBank and less of 1% divergence.
The sequence analysis makes this work the first report of Th.
ubiquitalis infected by L. (V.) lainsoni in Amazonas state.
N° 26 (1) 2015
109
TRANSMISSION OF LEISHMANIA INFANTUM TO
HAMSTERS THROUGH THE BITE OF
EXPERIMENTALLY INFECTED PHLEBOTOMUS
PERNICIOSUS AND MONITORING OF ANIMALS
INFECTIVITY BY XENODIAGNOSIS
PHYLOGENETIC ANALYSIS OF THE LEISHMANIA
ENRIETTII COMPLEX AND SPECULATIONS ON
THEIR VECTOR BIOLOGY
P.A. Bates, M.D. Bates, G. Kwakye-Nuako
Inés Martín-Martín, Maribel Jiménez, Estela González, Ricardo Molina
Lancaster University, UK
m.bates@lancaster.ac.uk
Keywords: Leishmania enriettii, phylogenetics, sand flies, midges
rmolina@isciii.es
Keywords: Phlebotomus perniciosus, Leishmania infantum, natural
transmission, xenodiagnosis
A leishmaniasis outbreak is currently occurring in south western
Madrid region (Spain) and already accounts for more than 560 human
cases. The causative agent is Leishmania infantum and the main vector
in the area is Phlebotomus perniciosus. All Leishmania isolates from
the focus have the same genotype as the strain
MHOM/ES/87/LOMBARDI but no association of isolate types and
virulence has been demonstrated. Our aim was to perform natural
transmission studies in animal models using isolates from P.
perniciosus collected in the field (IPER/ES/2012/BOS1FL1 and
IPER/ES/2012/POL2FL6) during the entomological survey carried
out in the transmission season of 2012 in the human leishmaniasis
outbreak area simultaneously with a well characterized L. infantum
strain (MCAN/ES/98/LLM-877).Two colonies of P. perniciosus
established from specimens captured in Madrid were experimentally
infected with 2.5x107 promastigotes/ml of L. infantum isolates. Ten
days after experimental infections, sand flies were fed on hamsters. In
addition, other hamsters were inoculated with 107 promastigotes/ml
of the same cultures. Effective transmissions by bite were monitored
and compared with intraperitoneal infections. Experimental
infections were successful but natural transmission by sand fly bite
was only achieved with isolates from the outbreak. Even attempts of
biting - without blood ingestion by infected sand flies with the focus
isolate - led to a transmission of the parasite. Infected hamsters were
xenodiagnosed every 2 months and were infective to sand flies as soon
as 2 months post-transmission experiments. Hamsters inoculated with
parasites from the outbreak presented clinical visceral disease and
were also infective to sand flies when they were tested by
xenodiagnosis. Vector transmission generated a slower progression of
clinical disease than intraperitoneal infection, but both were infective
to P. perniciosus by xenodiagnosis studies. A visceral leishmaniasis
model that mimic the natural conditions of transmission in nature was
set and allowed us to compare Leishmania strain virulence and
infectivity to sand flies. The differences found in the outcome of the
disease and infectivity to sand flies of infected hamsters by strains that
are circulating in the focus would contribute to the understanding of
their role in the epidemiology of the human leishmaniasis outbreak in
Madrid.This study supported by the Spanish Project AGL2008-01592
and by EU grant FP7-2011-261504 EDENext.
Recent work has provided evidence for a new phylogenetic grouping
of Leishmania parasites, the Leishmania. enriettii complex, that may
represent a new Leishmania subgenus in addition to Leishmania,
Viannia and Sauroleishmania. Known members of the L. enriettii
complex include L. enriettii itself (from Brazil), L. martiniquensis
(from Martinique, a Caribbean island) and L. australiensis (from
Australia). To these we now add further species from Namibia, Ghana
and Thailand. Some of these parasites are human pathogens, whilst
others appear to be confined to wild animals. The very wide
geographical spread of the L. enriettii complex, more widely
distributed than any of the existing subgenera, together with their
branching from the base of the Leishmania phylogenetic tree, both
suggest an early branching and ancient lineage. The vectors of the L.
enriettii complex have not been established with certainty for any
member. It has been speculated that L. enriettii has a phlebotomine
sand fly vector, however, this has not been proven, and L. enriettii
appears not able to establish infection in Lutzomyia longipalpis, a
widely used permissive sand fly host. For the other members there is
little evidence available, except for L. australiensis. In this species
evidence to date supports the vectors to be day-biting midges of the
genus Forcipmyia, although transmission by midge-bite has not been
attempted yet. It therefore appears that some, or maybe even all, of the
L. enriettii complex may have non-sand fly vectors. This speculation
remains to be proven or disproven, with efforts currently underway in
both Ghana and Thailand to identify potential vectors. However, if
true, this would require a re-definition of the genus Leishmania to
include non-sand fly transmission or the creation of a new genus to
accomodate the non-sand fly transmitted parasites.
N° 26 (1) 2015
VECTOR COMPETENCE OF NYSSOMYIA WHITMANI
(DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE) TO
LEISHMANIA (LEISHMANIA) AMAZONENSIS
N AT U R A L I N F E C T I O N O F L U T Z O M Y I A
(NYSSOMYIA) Y. YUILLI AND L.
( T R I C H O P H O R O M Y I A ) O C TAV I O I W I T H
LEISHMANIA SPP. IN AGUAS CALIENTES, LA
CONVENCION, CUSCO, PERÚ.
Magda Freitas Fernandes1, Kleiton Maciel dos Santos1, Antonio Carlos
Ferrari Júnior1, Ana Paula Silva Levay1, Fábio Juliano Negrão1, Wedson
Desidério Fernandes1, Elisa Teruya Oshiro2, Maria Elizabeth Moraes
Cavalheiros Dorval2, José Dilermando Andrade Filho3, Alessandra
Gutierrez de Oliveira2, Eunice Aparecida Bianchi Galati4
1
Universidade Federal da Grande Dourados (UFGD), Mato Grosso do Sul, Brazil.
2
Universidade Federal de Mato Grosso do Sul (UFMS), Brazil. 3Centro de Pesquisas
René Rachou, Fundação Oswaldo Cruz (Fiocruz), Minas Gerais, Brazil. 4Universidade
de São Paulo (USP), São Paulo, Brazil.
egalati@usp.br
110
D. Rado1, D. Pita-Pereira4, T. Araujo Pereira4, MM Quispe-Florez1, L.
Brahim3, E. Rangel3, B. Valladares2, R. Pacheco1
1
Instituto Universitario de Enfermedades Tropicales y Biomedicina del Cusco –
UNSAAC, Perú; 2Instituto Universitario de Enfermedades Tropicales y Salud Publica
de Canarias, Universidad de La Laguna, Tenerife, España; 3Laboratório de
Transmissores de Leishmanioses – Instituto Oswaldo Cruz/FIOCRUZ, Brasil;
4
Laboratório de Biologia Molecular e Doenças Endêmicas – Instituto Oswaldo
Cruz/FIOCRUZ, Brasil
dirc82@gmail.com
Keywords: Vector, Competence, Nyssomyia whitmani, Leishmania
amazonenses
Findings of phlebotomine species naturally infected by Leishmania
spp., and not yet implicated in the transmission of particular
leishmaniasis agents, bring out the need to investigate those species'
vectorial competence. The objective of this study was to investigate
the experimental susceptibility of Nyssomyia whitmani to Leishmania
(Leishmania) amazonensis and its transmission to uninfected
(susceptible) hamsters. Wild phlebotomine species were captured in a
forest fragment in Dourados, MS, during the night period with black
modified Shannon traps and the use of electrical aspirators. Males and
females were kept in cages for mating and the blood meal was taken on
susceptible hamsters exposed to biting for two hours. After 48 hours
the engorged females were placed in individual polyethylene vials for
egg laying. The eggs were then transferred to small Petri dishes until
the emergence of the adult specimens. The susceptibility tests with Ny.
whitmani F1 females were undertaken in two stages: in the first
experiment 156 specimens emerged in an average period of 55 days.
Of the 92 females, 63 (68.5%) took the blood meal on the infected
hamsters. Of those 63, 22 (34.9%) took the second meal on the
susceptible hamsters: 13 (20.6%) on the 5th day, 5 (7.9%) on the 6th
day, 2 (3.2%) on the 12th day and 2 (3.2%) on the 21st day. In the
second experiment 713 specimens emerged in an average period of 58
days. 561 females took the blood meal on the infected hamsters, and
131 (23.4%) the second blood meal on the susceptible hamsters: 13
(2.3%) on the 6th day, 82 (14.6%) on the 7th day, 6 (1.1%) on the 8th
day, 19 (3.4%) on the 9th day, 6 (1.1%) on the 10th day, 3 (0.5%) on the
12th day and 2 (0.4%) on the 14th day. A total of 27 susceptible
hamsters were exposed to the females which had fed themselves on
the infected hamsters. The confirmation of infection by Leishmania in
the hamsters exposed for the 2nd blood meal was undertaken by
Polymerase Chain Reaction to kDNA followed by Restriction
Fragment Length Polymorphism Assay with Hae III of the samples of
the spleen of the 27 hamsters, giving a 100% positive reaction. Seeing
that Ny. whitmani was experimentally infected by L. amazonensis and
succeeded in transmitting the infection to susceptible hamsters, its
vectorial competence for this parasite has thus been demonstrated .
However, it is necessary to undertake studies for the assessment of its
vectorial capacity in regard to the transmission of L. amazonensis.
Keywords: Natural Infection, L. y. yuilli , L. octavioi, Leishmania
spp.
Aguas Calientes (at 890 m) is an important endemic area of
leishmaniosis in the province of La Convencion (Cusco, Perú). The
most frequently species found in the area are L. y. yuilli (47,25%) and
L. octavioi (25,97%).With the aim of determine natural Leishmania
spp. infection in this species, we analized, by PCR multiplex, 550
females of . L. y. yuilli and 170 females of L. octavioi (grouped in
pools of 10 individuals) that come from collection made in intra, peri
and extra domestic environments with Shannon and CDC traps
during dry and rainy seasons in 2010-2011. Positive result were
detected in seven pools of L. y. yuilli (from extradomiciliary
collection) and three pools of L. octavioi (from intra, peri and
extradomiciliary collection) with a minimum infection rate of 1,27%
and 1,76%, respectively. In the studied area, both species could be
involved in Cutaneus Leishmaniosis epidemiology.Support by IOCFIOCRUZ, Brazil; CNPq, Brazil and AECID, Spain.
N° 26 (1) 2015
LUTZOMYIA (NYSSOMYIA) Y. YUILLI AND L.
(TRICHOPHOROMYIA) SP. FROM ALTO IVOCHOTE,
LA CONVENCIÓN, CUSCO, PERU
AGGREGATED PATTERN OF Leishmania infantum
chagasi TRANSMISSION FROM NATURALLY
INFECTED DOGS TO LABORATORY REARED
Lutzomyia longipalpis THROUGH XENODIAGNOSIS
Anita Quispe1, MA Quispe-Ricalde2, D. Rado1, MM Quispe-Florez1, M.
Soto1, Pérez JE3, B. Valladares2, R. Pacheco1
1
de Canarias, Universidad de La Laguna, Tenerife, España; 3Instituto de Medicina
Tropical Alexander von Humboldt, UPCH, Lima, Perú
anitaes001@gmail.com
Keywords: Leishmaniosis, Lutzomyia, PCR RFLP hsp70, natural
infection
Alto Ivochote is an endemic area for leishmaniosis and Lutzomyia spp.
in large populations are frequent in the area. The objective of this study
was to combine entomological with molecular techniques to
characterize Leishmania spp. detected in Lutzomyia (Nyssomyia) y.
yuilli y Lutzomyia (Trichophoromyia) sp., as they are the most
frequently species for the study area. From collection made during the
rainy and dry seasons of 2011, 594 females of Lu. (N.) y. yuilli and 458
females of Lu. (T.) sp., were studied to determine the natural infection
with Leishmania and species characterization through PCR – RFLP
hsp70. Both species were found naturally infected by Leishmania
(Viannia) sp. with natural infection rates of 18.35% and 3.57%,
respectively. On the other hand Leishmania (V.) braziliensis was
detected and characterized in 19 females of Lu. (N.) y. yuilli from
extra domestic collection with Shannon trap. These results suggest
that Lu. (N.) y. yuilli play an important role in the transmission cycle
of L. (V.) braziliensis to man and animals in Alto Ivochote.
111
Andre Antonio Cutolo1, Fredy Galvis Ovallos2, Cecilia Oliveira
Lavitschka2, Mariana Dantas da Silva2, Leticia Maria Molla3, Fabio dos
Santos Nogueira3, Vera Lucia Pereira-Chioccola1, Ingrid Menz3, Eunice
Aparecida Bianchi Galati2
1
Laboratorio de Biologia Molecular de Parasitas e Fungos, Instituto Adolfo Lutz, São
Paulo, Brazil. 2 Departamento de Epidemiologia, Faculdade de Saúde Pública, USP,
São Paulo, Brazil. 3 DVM Independent Researcher, São Paulo, Brazil
cutoloandre@yahoo.com
Keywords: Xenodiagnosis, Visceral Leishmaniasis, Vector
Competency, Transmission Rate
Thirty six Visceral Leishmaniasis symptomatic dogs, naturally
infected with Leishmania infantum chagasi were evaluated through
xenodiagnosis in order to check infectivity rate to laboratory reared
Lutzomyia longipalpis nulliparous females. Dogs were from an
American Visceral Leishmaniasis endemic area in São Paulo state,
Brazil, and confirmatory diagnosis for Leishmania infection was
performed through serology, cytology and molecular methods. All
animals were considered polysymptomatic at the time of
xenodiagnosis. Sandflies were dissected five days after bloodfeeding
under light microscopy in order to detect live Leishmania
epimastigotes and promastigotes in the gut contents. Sandfly
infection was scored as low (+), medium (++) and high (+++) level
depending on the number of protozoa observed. Among the 36 dogs
evaluated, 19 (52.8%) infected sandflies. An average of 22 insects
were dissected per exposed dog, with 805 sandflies dissected in total
and 99 (12.3%) found infected. Positive dogs had different levels of
infectivity to sandflies. Individual dog infectivity rates ranged from
4% (1/25) to 95% (19/20) of dissected sandflies. Fourteen (73.7%)
positive dogs had infectivity rates ranging from 4% to 20%, as
following: 4.0% (1/25), 4.8% (1/21), 5.0% (1/20), 5.0% (1/20), 5.0%
(1/20), 6.1% (2/33), 6.7% (1/15), 10.0% (2/10), 12.5% (2/16), 12.5%
(3/24), 14.3% (3/21), 15.0% (3/20), 19.1% (4/21) and 20.0% (4/20).
In addition, these 14 dogs generally infected sandflies at a low
intensity, with only one sandfly being scored as +++. Five (26.3%)
dogs were the most infectious animals with 60.0% (6/10), 65.0%
(13/20), 75.0% (15/20), 85.0% (17/20) and 95.0% (19/20) infected
sandflies per exposure, and these dogs also generally infected
sandflies at a higher intensity. Among the 99 infected sandflies, 29
(29.3%) came from the fourteen lowest infectious dogs and 70
(70.7%) sandflies originated from the five most infectious dogs. An
aggregated pattern of Leishmania infantum chagasi transmission
from dogs to sandflies was observed in this study. This finding
suggests that identification of the most infectious dogs within a
population and consequently use of proper transmission blocking
tools directed to them, may be an essential feature of transmission
control for this zoonotic visceral leishmaniasis causing agent in
Brazil.
N° 26 (1) 2015
ID 106-P Genome & Genetics
V E C TO R S O F L E I S H M A N I A M E X I C A N A I N
SOUTHEASTERN OF MEXICO
GENETIC CHARACTERIZATION OF NORTHERN
AND SOUTHERN POPULATIONS OF PHLEBOTOMUS
SERGENTI (DIPTERA: PSYCHODIDAE)
POPULATIONS IN MOROCCO
Pech-May Angélica1, Sánchez-García Laura2, Berzunza-Cruz Miriam2,
Becker-Fauser Ingeborg2, Escobedo-Ortegón Javier3, Rebollar-Téllez
Eduardo A4
1
Centro Regional de Investigación en Salud Pública -INSP, Tapachula, Chiapas,
México. 2Departamento de Medicina Experimental, Facultad de Medicina,
Universidad Nacional Autónoma de México, Hospital General de México, México,
D.F., México. 3Departamento de Enfermedades Infecciosas y Transmitidas por vector,
Centro de Investigaciones Regionales Dr. Hideyo Noguchi, Universidad Autónoma de
Yucatán, Mérida, Yucatán, México. 4Laboratorio de Entomología Médica,
Departamento de Zoología de Invertebrados, Facultad de Ciencias Biológicas,
Universidad Autónoma de Nuevo León, Monterrey, Nuevo León, México.
apechmay@gmail.com
In Mexico, leishmaniases represents a public health problem.
Leishmania (Leishmania) mexicana has so far been identified as the
main ethologic agent of cutaneous leishmaniasis (LCL) along the
country. The southeaster of Mexico is the main endemic area for LCL
and it has been demonstrated that Bichromomyia olmeca olmeca is the
primary vector, however Lutzomyia cruciata, Psychodopygus
panamensis and Psathyromyia shannoni have also been found
infected by PCR with Le. (Le.) mexicana. Therefore we suggested that
these species may be acting as primary and/or secondary vectors in
some areas of Yucatan Peninsula (YP), Mexico. Here we present a
resume of the state of the knowledge of the abundance, sandfly biting
rates and infection rate of Le. (Le.) mexicana vectors in YP. The results
of the abundance per species in each location indicated that the most
abundant species in the YP are Bi.o. olmeca (66%), followed by Pa.
shannoni (52%) and Ps. panamensis (43%). Regarding to the monthly
biting rate, the results showed that it depends of each species and
locality. The mean biting rate per hour, showed peak from 18:00 to
19:00 in all localities. The high infection rate was in 20JUN (5.3%),
the sandfly Pa. shannoni was infected in all localities, while that only
sandfly Bi. o. olmeca was infected in collections made in Shannon and
Disney traps. We can propose according with the sand fly seasonal
abundance distribution and the biting rates, those biotic and abiotic
factors, which vary within a period time, affects sand fly populations.
112
Boussaa Samia1,2; Ouanaimi Fouad1; Kahime Koloud1; Echchakery
Mohamed1; Boumezzough Ali1
1
Equipe d'Ecologie Animale-Environnement, Faculté des Sciences Semlalia,
Université Cadi Ayyad, Marrakech, Maroc; 2ISPITS-Institut Supérieur des Professions
Infirmières et des Techniques de Santé, Ministère de Santé, Marrakech, Morocco
fouad.ouanaimi@yahoo.fr
Keywords: Phlebotomus sergenti, Isoenzyme, Mitochondrial DNA,
Morocco
In Morocco, Phlebotomus sergenti is the proven vector of
Leishmania tropica, a causative agent of cutaneous leishmaniasis
(Guilvard et al., 1991). Many Moroccan L. tropica foci have been
reported, in the rural areas of Azilal, Essaouira and Chichaoua
(Pratlong et al., 1991; Guernaoui et al., 2005; Ramaoui et al., 2008),
but also in urban and peri-urban areas of Taza and Zouagha Moulay
Yaacoub (Guessous-Idrissi, 1999; Rhajaoui et al., 2004). The taxon
known as P. sergenti shows considerable genetic variation over its
large global area of distribution (Depaquit et al., 2002; Yahia et al.,
2004; Moin-Vaziri et al., 2007). This study presents a genetic
comparison, using isoenzyme and molecular tools, between northern
(Chefchaouene, Ouezzane, and Taounate) and southern (Marrakech,
Ourika, Rhmate and Touama) populations of Moroccan P. sergenti.
For morphological studies sandflies were collected on sticky-paper
traps. After dissection, sandflies were slide-mounted in Marc Andre
solution and examined by light microscopy. For isoenzyme and
molecular analyses, specimens were collected in CDC light traps and
immediately stored at -80°C. Ten isoenzyme systems were tested and
the qualitative analysis of zymogram profiles revealed nine
polymorphic loci (HK, PGM, PGI, 6PGD, FUM, MDH1, ME, ICD2
and ACO). A section of the mitochondrial gene Cytochrome b (Cyt b)
was amplified by PCR with the primer pair CB3-FC/CB-R06. The
calculation of allele frequencies, tests for deviation from
Hardy–Weinberg equilibrium, and phenetic analysis were performed
using Genepop and Biosys2 programs. Our results showed that these
two P. sergenti populations do not have the genetic characteristics of a
single species, as indicated by the HW disequilibria for several tested
loci and by the presence of two P. sergenti mitochondrial lineages.
The epidemiological significance of this genetic variability merits
future investigation. References: Depaquit et al., 2002. Int J
Parasitol, 32, 1123–1131. Guernaoui et al., 2005. J M Entomol, 42,
697–701. Guessous-Idrissi, 1999. Recherche Nationale, les Cahiers
du Médecine, 2, 37–40.Guilvard et al., 1991. Ann Parasitol Hum
Comp, 66, 96–99. Moin-Vaziri et al., 2007. Acta Trop, 102,
29–37.Pratlong et al., 1991. Ann Parasitol Hum Comp, 66, 100–104.
Ramaoui et al., 2008. Parasitol Res, 103, 859–863. Rhajaoui et al.,
2004. Trans R Soc Trop Med Hyg, 98, 299–301. Yahia et al., 2004.
Parasite, 11, 189–199.
N° 26 (1) 2015
113
ID 107-P Genome & Genetics
ID 108-P Leishmania - Sand fly Interactions
EVIDENCE FOR GENETIC DIFFERENTIATION AT
THE MICROGEOGRAPHIC SCALE IN
PHLEBOTOMUS PAPATASI POPULATIONS FROM
SUDAN
ANALYSIS OF CARBOHYDRATE EPITOPES ON THE
S U R FA C E O F M I D G U T E P I T H E L I U M O F
LUTZOMYIA (LUTZOMYIA) LONGIPALPIS AND
LUTZOMYIA (NYSSOMYIA) ANTUNESI (DIPTERA,
PSYCHODIDAE) FROM PARÁ, STATE AMAZONIAN
BRAZIL
Noteila M Khalid, Marium A Aboud, Fathi M Alrabba, Dia-Eldin A
Elnaiem and Frederic Tripet
Department of Zoology, Khartoum College of Medical sciences, Khartoum, Sudan.
Noteila@hotmail.com
Keywords: Phlebotomus papatasi, Sudan, Gene flow, Genetic
differentiation.
Background: Cutaneous Leishmaniasis (CL) is endemic in Sudan. It is
caused by Leishmania major parasites and transmitted by
Phlebotomus papatasi sandflies. Recently, uncommon clinical
manifestations of CL have been reported. Moreover, L. donovani
parasites that cause Visceral Leishmaniasis (VL) have been isolated
from CL lesions of some patients who contracted the disease in
Khartoum State, Central Sudan with no history of travelling to VL
endemic sites on south-eastern Sudan. Because different clinical
manifestations and the parasite behaviour could be related to genetic
differentiation, or even sub-structuring within sandfly vector
populations, a population genetic study was conducted on P. papatasi
populations collected from different localities in Khartoum State
known for their uncommon CL cases and characterized by contrasting
environmental conditions. Methods: A set of seven microsatellite loci
was used to investigate the population structure of P. papatasi samples
collected from different localities in Khartoum State, Central Sudan.
Populations from Kassala State, Eastern Sudan and Egypt were also
included in the analyses as outgroups. The level of genetic diversity
and genetic differentiation among natural populations of P. papatasi
was determined using FST statistics and Bayesian assignments.
Results: Genetic analyses revealed significant genetic differentiation
(FST) between the Sudanese and the Egyptian populations. Within the
Sudanese P. papatasi populations, one population from Gerif West,
Khartoum State, exhibited significant genetic differentiation from all
other populations including those collected as near as 22 km.
Conclusion: The significant genetic differentiation of Gerif West P.
papatasi population from other Sudanese populations may have
important implication for the epidemiology of leishmaniasis in
Khartoum State and needs to be further investigated. Primarily, it
could be linked to the unique location of Gerif West which is confined
by the River Nile and its tributaries that may act as a natural barrier for
gene flow between this site and the other rural sites. The observed high
migration rates and lack of genetic differentiation among the other P.
papatasi populations could be attributed to the continuous human and
cattle movement between these localities.
Oliveira, D.M.S.1; Lima, J.A.N2.; Silva, B.J.M 1; Farias, L.H.S1; Santos,
T.V2.; Silveira, F.T2; Silva, E.O1
¹Laboratório de Biologia Estrutural/Laboratório de Parasitologia, Instituto de
Ciências Biológicas, Universidade Federal do Pará, Brazil.2 Instituto Evandro Chagas
- Ministério da Saúde
davimso@yahoo.com.br
Key-Words: Helix pomatia lectin, Lutzomyia, glycoprotein.
Leishmaniasis
Leishmaniases are worldwide diseases that occur in 98 countries. In
Brazil, cases of American Visceral Leishmaniasis (AVL) and
Cutaneous Leishmaniasis (CL) has been registrated in all Federate
Units, including Pará State. Lutzomyia (Lutzomyia) longipalpis is
considered to be the main vector of Leishmania (Leishmania)
infantum chagasi, the etiological agent of AVL while Lutzomyia
(Nyssomyia) antunesi is the proven vector of Leishmania (Viannia)
lindenbergi suspected of causing CL. The specificity of the parasitesandflies interaction has to do with leishmania surface molecules as
lipophosphoglycan (LPG) which is recognized by receptors of
midgut epithelium. However, in permissive sand flies as L.(L.)
longipalpis, the interaction is mediated by expressions of
glycoproteins containing N-acetylgalactosamine (GalNAc) of
epithelial insect midgut which probably acts as recognition sites by
molecules lectin-like which are present on the Leishmania surface.
The aim of this work was to compare the midgut expression of
glycoprotein containing GalNAc of L. (L.) longipalpis from a close
colony to the glycoprotein expression of field samples of L. (L.)
longipalpis and L. (N.) antunesi of Cametá Municipality, Pará State.
Phlebotomines were dissected into 4% paraformaldehyde and each
individual midgut was sectioned longitudinally; then incubated with
FITC conjugated Helix pomatia lectin (HPA-FITC) and
Concanavalin A (Con-A-FITC) which are specific for GalNAc and
manose residues, respectively. Lysates of seven midguts of each
phlebotomine species from colony and field were analyzed by SDSPAGE followed by western blotting with HPA and Con-A for
detecting glycoprotein containing GalNAc and manose. In both
colony and field samples, HPA-FITC and Con-A-FITC showed
positive reactivity suggesting the presence of both GalNAc and
manose. SDS-PAGE followed by western blotting showed that
midguts lysates were GalNAc- and manose-positive for displaying
peptides ranging from 35 to 75 kDa and 51 kDa that bind to HPA and
Con-A, respectively. The results indicated that glycoprotein
containing GalNAc occurs in both species from areas endemic for
AVL and CL of Cametá municipality.
Supported by CAPES, CNPq/UFPa, CNPq/MCT/CT-INFRA/CTPETRO (Process number 620179/2008),
MCT/CNPq/FNDCT/PROCAD-NF CAPES/FAPERJ and
IEC/SVS/MS.
N° 26 (1) 2015
COMPARATIVE ANALYSIS BETWEEN CANINE
VISCERAL LEISHMANIASIS AND SANDFLY
PRESENCE IN THE ENDEMIC MUNICIPALITIES
SURVEY OF SANDFLY VECTORS IN AREAS
S U R R O U N D I N G P E D R A B R A N C A M A S S I F,
MUNICIPALITY OF RIO DE JANEIRO, BRAZIL
Denise Maria Bussoni Bertollo, Regiane de Cassia Hilário Castilho,
Márcia Maria Costa Nunes Soares, Juliana Kindler Figueiredo, Brígida
Helena da Silva Meneghello; Roberto Mitsuyoshi Hiramoto; José
Eduardo Tolezano
Quezia Nunes Peres Dias, Marcos Barbosa de Souza, Shênia Patrícia
Corrêa Novo, Antônio de Medeiros Meira e Cesar do Santo Ponte
Instituto Adolfo Lutz
114
Fundação Oswaldo cruz, Fiocruz / Escola Nacional de Saúde Pública Sérgio Arouca,
ENSP.
mabaza@ensp.fiocruz.br
dmbbertollo@ial.sp.gov.br
Keywords: Leishmaniasis, Sandfly, entomological investigations
Visceral Leishmaniasis (VL) is an infectious disease considered a
zoonosis that infects humans as incidental hosts. VL has experienced
major changes in the pattern of transmission, initially restricted to
rural and peri-urban environments and, more recently, reaching
important urban centers. The trypanosomatid protozoan species
Leishmania infantum (chagasi) the etiological agent that causes VL,
being transmitted by the hematophagous sandfly vector Lutzomyia
longipalpis. The objective of this study was to correlate the occurrence
of Canine Visceral Leishmaniasis (CVL) and the presence of vectors
in municipalities in the northwestern region of São Paulo State, in the
period 2008-2014. During the period, 38 municipalities were
investigated, in 11 (28.9%) both infected dogs and sandfly vectors
were recorded. In 25 (65.7%) only vectors were found and recently 2
human cases were diagnosed. Adults sandflies were predominant in
peridomestic environments during entomological investigations in
the municipalities with vector. Such environments were characterized
by the presence of pets and appropriate habitats for breeding of L.
longipalpis in urbanized area. Actions to reduce the probable sites of
vector propagation have been performed by means of environmental
management in these municipalities. Climatic factors may determine
the epidemiology of VL and the adaptation and gradual migration of
vectors to urban regions. Factors such as temperature, humidity,
rainfall and presence of sandflies must be studied and understood in
order to reduce the spread of vector populations and consequent
increase in disease transmission.
Keywords: Leishmaniasis, Sandfly fauna, Pedra Branca Massif,
Municipality of Rio de Janeiro
Introduction: Several sand fly species have been registered with
natural infection with Leishmania spp. In Brazil, American
cutaneous leishmaniasis (ACL) is worthy of attention due to its high
incidence and risk of deformities in humans. The incriminated
vectors of ACL are Nyssomyia intermedia, Migonemyia migonei,
Nyssomyia whitmani, Bichromomyia falviscutellata, Nyssomyia
umbratilis and Psychodopygus wellcomei. American visceral
leishmaniasis (AVL) has been expanding rapidly in urban areas,
having as its principal vectors Lutzomyia longipalpis and Lutzomyia
cruzi. Materials and Methods: This study was conducted in 31
endemic areas in the surroundings of Pedra Branca Massif, to the
west of the city of Rio de Janeiro, Brazil: A. Vasconcelos, Barra de
Guaratiba, Serra dos Caboclos, Cabuçu, Cachamorra, Camorim,
Carapiá, Colônia Juliano Moreira, Estrada do Magarça, Estrada do
Monteiro, Estrada das Taxas, Estrada dos Teixeiras, Grota Funda,
Grumari, Guandu do Sena, Ilha de Guaratiba, Jardim Novo
Realengo, Lameirão Pequeno, Maciço do Mendanha, Pau da Fome,
Serra de Piabas, Rio Bonito, Santa Maria, Serra do Barata, Serra do
Bangu, Serra do Rio da Prata, Serra de Santíssimo, Serra do Viegas,
Vargem Grande, Vargem Pequena and Vila Kennedy. Sand fly
collections were conducted during two years (2003-2005) in
peridomestic habitats using CDC light traps, targeting the sand fly
habitats in these regions. The collections were performed from 6 pm
to 7 am the following day and specimens were analyzed in the
laboratory and identified to species level. Results: A total of 46,971
specimens belonging to 13 species were collected. N. intermedia, M.
migonei and L. longipalpis were the most prevalent species with a
total of 40,449; 4,072 and 1,750 specimens, respectively. The months
with highest sand fly density were June, July, September and March.
Discussion and Conclusion: The high density of the three most
common species can be explained by the presence of hen houses in
these localities, corroborating the literature on studies conducted in
different localities. This survey enabled us to discriminate and
quantify the species by regions, contributing to eco-epidemiological
data about vector, once the risk of transmission of the parasite
depends on the existence of a large number of vector species, and also
the emergence of disease previously restricted to the sylvatic species.
N° 26 (1) 2015
ID 111-P Physiology and Colonization
ID 112-P Physiology & Colonization
ROTATION OF THE EXTERNAL GENITALIA IN MALE
PHLEBOTOMINE SANDFLIES (DIPTERA,
PSYCHODIDAE) IN LABORATORY CONDITIONS
AND IN CAPTURED SPECIMENS IN ALGARVE,
PORTUGAL
SOME FIELD AND LABORATORY OBSERVATIONS
ON THE BIOLOGY OF NYSSOMYIA ANTUNESI
(DIPTERA: PSYCHODIADAE), A SUSPECTED
VECTOR OF LEISHMANIA (VIANNIA) LINDENBERGI
IN AMAZONIAN BRAZIL
Ferrolho J.1, Gomes J.1, Alves-Pires C2, Cristóvão J.M.1, Maia C.1,3,
Campino L.1,2, Afonso M.O.1,2
Thiago Vasconcelos dos Santos1, Marliane Batista Campos1, José Aprígio
Nunes Lima1, Roberto Carlos Feitosa Brandão1, Fábio Márcio Medeiros
da Silva1, Maria Sueli Barros Pinheiro1, Luciene Aranha da Silva Santos1,
Edna De Freitas Leão1, Iorlando da Rocha Barata1, Fernando Tobias
Silveira1
1
UEI Parasitologia Médica, Instituto de Higiene e Medicina Tropical, Universidade
Nova de Lisboa (IHMT-UNL), Rua da Junqueira 100, 1349-008 Lisboa, Portugal;
2
UPMM/FCT, IHMT-UNL; 3CMDT/FCT, IHMT-UNL
joana.ferrolho@ihmt.unl.pt
115
1Laboratório de Leishmanioses “Prof. Dr. Ralph Lainson”, Seção de Parasitologia,
Instituto Evandro Chagas (SVS/ MS), Ananindeua, Pará, Brazil;
Keywords: Phlebotomine sandflies, genitalia, Portugal
Phlebotomine sandfly females are responsible for the transmission of
several pathogens, such as Leishmania spp., affecting humans and
animals in the Old and New World. Recently, ecological and climatic
changes have been associated with an increase of vectorial density and
leishmaniasis expansion. Phlebotomus perniciosus and P. ariasi are
the proven vectors of L. infantum in Portugal. Phlebotomine males
eclose from the pupae with un-rotated external genitalia. During the
initial 16-24 hours of adult life time, the external genitalia has to
undertake a complete rotation on the longitudinal axis through 180° to
assume a mature position. Recording the physical characteristic of
males with unpartially or completely rotated external genitalia can
help to predict the breeding sites and the proximity of adult
movements. In the present study, we firstly investigated the timing of
complete rotation of the male external genitalia in laboratory
conditions. P. perniciosus males were used as a model and were
obtained from a previously established colony at the IHMT-UNL and
maintained with optimal rearing conditions throughout the assay.
After eclosion, males were separated and observations were carried
out at 24 and 48 hours. At each sampling time point, external genitalia
were observed under a stereomicroscopic, either in vivo or after
preservation in 90% ethanol. Secondly, we identified morphologically
the sandfly male species captured from May to November 2013 in the
Sotavento and Barlavento Algarvio regions, Portugal, in different
biotopes types (domestic and peridomestic) using CDC miniature
light and sticky oil paper traps. Finally, we characterized the external
genitalia rotation in the captured males and investigated the relation
between the species and types of traps, periods of capture and natural
biotopes/collecting places. In this work, we report the timing for
external genitalia to complete rotation under laboratory conditions in
P. perniciosus males, so far the only confirmed vector of L. infantum in
Algarve, and based upon this model, we related the proximity of the
captured sandfly species with their breeding sites in the selected
Algarve regions. More information on sandfly breeding sites will
facilitate their control by source reduction and host protection,
especially in urban and periurban areas. This work was partially
supported by EDENext (FP7-INCO-CT-2010-261504).
C. Maia (SFRH/BPD/44082/2008) holds a fellowship from FCT,
MCTES, Portugal
Keywords: Nyssomyia antunesi, Leishmania (Viannia) lindenbergi,
cutaneous leishmaniasis.
In the Amazon region, the phlebotomine sand fly Nyssomyia antunesi
Coutinho, 1939 is recognized to be the main suspected vector of
cutaneous leishmaniasis (CL) caused by Leishmania (Viannia)
lindenbergi Silveira et al. 2002, with basis on the anthropophilic
behavior and high frequency in endemic areas under the occurrence of
the disease. Despite the epidemiological relevance, little is known
regarding the biology of this sand fly. Thus, this study aimed to
describe some field and laboratory observations of Ny. antunesi in
order to provide further evidences on the ecology of CL in this region.
During the Amazonian dry season (September-October/2013), an
entomological survey was conducted in a forest fragment located
nearby Maguari River margin (1º20'21''S 48º22'13''W) in Ananindeua
municipality, Pará State, Brazil. This location was chosen based on a
previous human case of CL attended at the Laboratório de
Leishmanioses “Prof Dr. Ralph Lainson”, Instituto Evandro Chagas,
Ananindeua, Brazil. Collects were performed for 15 nights using
CDC light traps at 1,5m ground (n=6) and 20m at canopy (n=2). The
collected sand flies were identified and the females were dissected to
search flagellate infection. In addition, some of collected sand flies
were kept alive to be fed on hamster and the matrixes were prepared
individually or in groups of 50 individuals for oviposition. Immature
emerged stages were maintained with a diet based on a mixture of
triturated substrate and powdered beef liver (Difco). A total of 2.414
sand flies were collected and identified, belonging to 13 species. Ny.
antunesi accounted 83,7% of collected at 1,5m and 91,3% at 20m.
Two dissected females of this species were found harboring
flagellates morphologically distinguishable from Leishmania spp.
Out of approximately 2000 non-identified individuals kept alive, 70%
fed on hamster. Individualized females did not succeed for
oviposition. Two groups of 50 females generated F1 and achieved
until adult form in approximately 52 days with the following duration
average of immature stages: egg (10 days), L1 (9 days), L2 (11 days),
L3 (9 days), L4 (7 days), pupa (6). All the adults emerged were
confirmed as Ny. antunesi. However, the F2 did not continued
successfully. These results together with previous data corroborate to
indicate Ny. antunesi as potentially involved on the transmission of L.
(V.) lindenbergi in the Amazon region. Under laboratory conditions,
immature stages of Ny. antunesi seem to be well adapted for
colonization, however, survival of adult form should be more
carefully analyzed.
N° 26 (1) 2015
ATTRACTIVENESS OF DIFFERENT DIETS FOR
NYSSOMYIA NEIVAI (DIPTERA: PSYCHODIDAE)
LARVAE
EFFECT OF NYSSOMYIA NEIVAI (DIPTERA:
PSYCHODIDAE) DENSITY ON OVIPOSITION
1
2
Thais Marchi Goulart ; Vicente Estevam Machado ; Mara Cristina
Pinto2
1
Instituto de Biologia, Departamento de Biologia Animal-UNICAMP, Brazil;
Faculdade de Ciências Farmacêuticas-UNESP, Brazil
116
Thais Marchi Goulart1; Vicente Estevam Machado2; Mara Cristina
Pinto2
1
Instituto de Biologia; Departamento de Biologia Animal-UNICAMP; 2Faculdade de
Ciências Farmacêuticas-UNESP
2
thamarchi@gmail.com
thamarchi@gmail.com
Keywords: colony, density, oviposition
Keywords: Sand fly colony, immature stages, attractiveness of sand
fly diet
The necessity of improving techniques for sand flies colonization is
an important demand for further studies on vector biology and its
interaction with Leishmania spp. The crucial limitation for rearing
these insects is the high mortality during immature phases. Although
there are some studies focused on diets for sand fly larvae, this aspect
is still an issue for sand fly colonies. The aim of this research was to
evaluate the attraction of immature stages of Nyssomyia neivai for
different diets. Sand flies were collected in Santa Eudóxia, SP - Brazil
(edges of Mogi Guaçu river) using automatic light traps and manual
aspiration. At the laboratory, sand flies were maintained in cages
covered with "voil" (30x30x30cm) at 26 ± 1°C, 80–90% humidity,
12:12 (L:D) photoperiod with access to a piece of cotton soaked in
30% sucrose solution. The non-bloodfed field-collected females were
exposed to mouse as blood source. The bloodfed females were
transferred to an oviposition chamber. Fifty eggs of N. neivai were
placed in the center of a 150mL polystyrene container filled with 2 cm
of plaster of Paris. The plaster of Paris was divided in four quadrants
and over each one a specific diet was added: diet 1 (rabbit feces, rabbit
food, fish food and soil), diet 2 (rabbit feces and soil), diet 3 (rabbit
feces, soil and fish food) and diet 4 (soil and mud). The number of
larvae on each quadrant was observed daily. The containers were
maintained in the dark at 26 ± 1°C, 80–90% humidity. The experiment
was replicated four times. The results were analyzed by analysis of
variance (ANOVA) and Tukey test. According to the result, the most
attractive diet was diet 1, followed by diet 3. Both diets contain fish
food. Differences were considered significant among the four diets
(p<0.0001) but with no significant difference were verified between
diets 1 and 3 (p>0.05) or 2 and 4 (p>0.05). The presence of fish food
appears to be important on the attractiveness of the larvae stages. This
information is interesting for further studies on sand flies larvae
attractiveness.
The maintenance of sand flies colonies is a meticulous activity and
each species seems to have special requirements. Nyssomyia neivai is
an important vector of cutaneous leishmaniasis in South America and
data about its colonization are scarce. The aim of this study was to
evaluate females of N. neivai density effects on oviposition. Sand flies
were collected in Santa Eudóxia, SP - Brazil from 6 p.m. to 11 p.m.
using Castro aspirators. At the laboratory sand flies were maintained
in holding cages at 26 ± 1°C, 80-90% humidity, 12:12 (L:D)
photoperiod with access to a piece of cotton soaked in 30% sucrose
solution. The non-bloodfed field-collected females were exposed to a
mouse as blood source. Bloodfed females were selected and
transferred in groups with equal numbers of males to 250mL
polystyrene oviposition chambers. The cover of the chamber was
replaced with “voile” and the bottom filled with 2cm of plaster of
Paris as a substrate for oviposition. The sand fly couples were placed
in seven groups: 1, 5, 10, 15, 20, 25 and 30. The experiment was
replicated four times. The chambers were observed daily to check the
humidity, change of sucrose solution, mortality of gravid females and
presence of eggs. After the egg counting, dead sand flies were
removed and identified. The chambers were maintained in the dark at
26 ± 1°C, 80-90% humidity. The results were analyzed by analysis of
variance (ANOVA) and Tukey test. As result, all the females were
identified as N. neivai. The mean of eggs per female was highest when
sand flies were in groups than individualized, but the differences were
not statistically detected. The range of the means varied from 37.3 (±
5.3) eggs in chambers with only one female to 56.5 (± 3.9) eggs in
chambers with 25 females. When the number of couples in the
chamber is highest than 20, the females lay eggs on the top, “voile”
and wall of the chamber and due to this fact, these eggs dry and die.
The results obtained with N. neivai using groups of females indicated
that there is an increase on oviposition when females are in groups,
but considering all the conditions for the process, the ideal number of
couples for this species in the container of 250mL is 20.
N° 26 (1) 2015
ID 116-P Other Topics
ROLE OF AQUAPORINS IN THE PHYSIOLOGY OF
LEISHMANIA VECTOR LUTZOMYIA LONGIPALPIS
LEISHMANIASIS VIRTUAL LABORATORY (LVL) - A
NEW TOOL FOR THE STUDY OF AMERICAN SAND
FLIES (DIPTERA, PSYCHODIDADE,
PHLEBOTOMINAE)
Rita Mukhopadhyay1,
Goutam Mandal1, Jose Orta1, Srotoswoti
1
Mandal , Claudio Meneses2, Hamide Aslan Suau2, Eric Beitz3, Jesus
Valenzuela2, Marcelo Ramalho-Ortigao4, Shaden Kamhawi2
1
Department of Cellular Biology and Pharmacology, Herbert Wertheim College of
Medicine, Florida International University, Miami, USA; 2Vector Biology, National
Institute of Allergy and Infectious Diseases, Baltimore, USA; 3Dept. of Pharmaceutical
and Medicinal Chemistry Pharmaceutical Institute, University of Kiel, Germany;
4
Department of Entomology, Kansas State University, Kansas, USA.
rmukhop@fiu.edu
Keywords: Leishmania, aquaporins, Lutzomyia, sand fly
Introduction: Aquaporins (AQP) are the route of entry of water and
uncharged solutes including gases (carbon dioxide and ammonia) into
cells from bacteria to humans. They play highly significant roles from
cell migration in mammals to fecundity in Drosophila. Female
phlebotomine sand flies are vectors of Leishmania.sp worldwide.
Sand flies survive in widely variant climatic conditions of extremely
humid tropics to the deserts of the Middle East. Upon blood meal
ingestion, a large number of events are induced, including digestion,
metabolism, diuresis and ultimately oogenesis. Sand flies also ingest
plant sap or aphid and coccid honeydew rich in sucrose between blood
meals. Therefore, their gut has to deal with huge volumes of
fluids/water and a wide range of osmolarity. On the other hand, in
nature they oviposit on substrates ranging from decaying organic
matter to animals feces. How do they home in to these various sources
of feed either for themselves or for their young? There is a substantial
gap in understanding the underlying molecular mechanisms of these
physiological processes of the sand flies. We hypothesize that
aquaporins (AQPs) are at the center of these primary sand fly survival
activities. Yet AQPs have not been reported from this highly important
disease vector. Methods: We cloned five Lutzomyia longipalpis
(LULO) AQPs by RACE-PCR and expressed them in Xenopus
oocytes. Additionally, various tissues under different conditions were
isolated from the flies, and different AQP status was determined by
qPCR. Results: All AQPs facilitate water uptake in Xenopus oocytes.
Modulation of AQPs was also observed based on the feeding status,
tissue specificity and Leishmania infection. LULO-AQP1-4 are also
transcriptionally active in antennae and brain. Discussion: AQPs are
involved in major physiological processes of Lutzomyia. We also
propose that they are involved in homing behavior which is novel in
insect research.
117
Felipe Dutra Rêgo; Israel de Souza Pinto, Caroline Amaral Machado,
Gabriel Eduardo Melim Ferreira, Elisa Cupollilo, Paloma Helena
Fernandes Shimabukuro
Centro de Referência Nacional e Internacional para Flebotomíneos/Centro de
Pesquisas René Rachou/FIOCRUZ
felipedutra@cpqrr.fiocruz.br
Keywords: Barcode, Cloud computing, Phlebotominae
Cloud computing has becoming increasingly popular among
different kinds of internet services available to users, and it involves
platforms or applications that offer services remotely through the
web. Cloud-based services can be central to scientific advancements,
challenging the sustainability of an approach to distributed
computing rapidly being taken over by events and increased demands
from user needs in the past 15 years. The EU-Brazil Cloud Connect
(EUBrazilCC) project is built on a close collaboration between
European and Brazilian institutions working towards providing a
user-centric, real test bench for European and Brazilian research
communities to test the execution of scientific applications. This
project is being developed with three scientific use cases covering
multidisciplinary areas, such as biodiversity, epidemiology, public
health, and climate change. These scientific scenarios all require
complex workflow pipelines and access to huge datasets. One of the
scientific use cases is the Leishmaniasis Virtual Laboratory (LVL), a
collaboration between the Instituto de Salud Carlos III (WHO-CCL
Leishmania Collection) in Spain and FIOCRUZ in Brazil with the
collections of Leishmania (CLIOC) and sand flies (COLFLEB). The
aim of this work is to present the LVL use case to the sand fly research
community and to provide a pipeline to perform molecular analyses
of sand flies. At COLFLEB, our aim is to study the phylogenetic
relationships of American sand flies, including the main vectors of
Leishmania in Brazil (Nyssomyia intermedia, N. neivai, N. whitmani,
N. umbratilis, Bichromomyia flaviscutellata, Psychodopygus spp.,
and Lutzomyia longipalpis). Using cytochrome oxidase subunit I
(coI), our pipeline will work with uploaded sequence data from sand
flies generated by user or downloaded from public databases (e.g.
Barcode of Life Database - BOLD or GenBank). Analysis of data
through the pipeline will include: DNA sequence editing and
alignment, maximum likelihood analysis, Bayesian inference
analysis and tree editing.
N° 26 (1) 2015
IMMUNOLOGICAL STUDIES ON THE CUTANEOUS
LEISHMANIASIS IN AL-KHARJ REGION, SAUDI
ARABIA
SANDFLIES AND POPULAR KNOWLEDGE OF
LEISHMANIASIS TRANSMISSION IN A PERI-URBAN
AREA OF THE MUNICIPALITY OF SÃO GONÇALO
DO AMARANTE, BRAZIL
Mohammad S. Al-Khalifa; Ibraheem M Al-Hazza and M.
Y. Lubbad
Zoology Department, College of Science, P. O.Box 2455, King Saud University, Riyadh
11451, Saudi Arabia.
mkhalifa@ksu.edu.sa
Keywords: Cutaneous Leishmaniasis, Phlebotomus, Saudi Arabia,
Zoonosis.
Present study is an attempt to determine the nature of immunological
cells and the cytokine they produce in zoonotic cutaneous
leishmaniasis (ZCL) patients in an endemic zone, Al-Kharj area,
southeast of Riyadh Saudi Arabia. ELISA and the short-term culture
were the techniques used for this purpose. ELISA determines the level
of cytokines in the peripheral blood in ZCL patients. There were no
variations in the levels of IL-4 in the blood of ZCL patients before and
after treatment by tropical application of Pentostam. However, level of
cytokine IL-6 and IL-10 and TNF-α have significantly increased in the
patient's blood. Short-term cultures were used to study the level of
intra cellular cytokine and it observed that level of intra cellular IL-2
has significantly increased before the treatment compared to the same
patient after the Pentostam treatment. However, its level increased
when examined in patients after the three months treatment and there
were no variation in the levels of IL-4 in ZCL patients, before and after
treatment.
118
Maria de Lima Alves, Maria de Fátima Freire de Melo Ximenes, Marcos
Paulo Gomes Pinheiro, Tamy Elicia da Silva Lopes, João Batista da Silva
Junior
Universidade Federal do Rio Grandedo Norte - UFRN. malimalves11@gmail.com
Keywords: Popular knowledge, Phlebotomine sandflies, health
education
The peri-urban areas of cities are of particular interest to researchers,
especially in the field of public health, due to the socioeconomic,
environmental, sanitary and cultural characteristics of these regions,
which are unfavorable for public and collective health. In Rio Grande
do Norte, leishmaniases occur in different geographic regions of the
state both separately and concomitantly. The number of cases of
Visceral Leishmaniasis (VL) is higher than that of American
Cutaneous Leishmaniasis (ACL). The aim was to characterize the
community as to social, sanitary and environmental aspects as well as
the knowledge of residents regarding the transmission cycle of
leishmaniases in the Guanduba community located in the peri-urban
area of the municipality of São Gonçalo do Amarante. The
methodology consisted of applying a questionnaire containing
twenty semi-structured questions on epidemiological, sanitary and
environmental aspects involved in the transmission cycle of
leishmaniasis to 115 families enrolled in the Family Health Program
(PSF), Ministry of Health, in July 2013. The results demonstrate that
the Guanduba community suffers from socioeconomic problems, the
absence of urban and sanitary infrastructure as well as a number of
environmental problems. With respect to the disease process, 66% of
those interviewed did not know how leishmaniasis was contracted,
52% were unaware of how it was transmitted, 96% did not know how
to prevent it and 95% did not know the name of the transmitting
insect of leishmaniases. In the comunity was held on systematic
capture of sandflies with CDC light traps, which demonstrate the
occurrence of species incriminated as vectors of LV and LTA. The
catches indicate the presence of Evandromyia lenti 84%,
Evandromyia evandroi 8%, Lutzomyia longipalpis 6%, Sciopemyia
sordellii 1%, Evandromyia walkeri 0,5% e Nyssomyia intermedia
0,5%. It is concluded that the level of knowledge regarding
epidemiological cycles is weak and inadequate for prevention
measures, to the detriment of the accompaniment of these families by
the PSF in this community, where sanitary conditions, environmental
and entomological will increase the risk of disease transmission, as
evidence the presence of vector species of leishmaniasis. Thus, it is
recommended that educational measures being implemented by the
Family Health Strategy in partnership with other public institutions.
N° 26 (1) 2015
119
BOOKLET EDUCATIONAL IN COMIC STRIP ABOUT
SANDFLIES AND THE LEISHMANIASIS IN THE
STATE OF RIO GRANDE DO NORTE, BRAZIL
Maria de Lima Alves, Maria de Fátima Freire de Melo Ximenes,
Magnólia Florêncio Fernandes de Araújo, Marcos Paulo Gomes
Pinheiro, Tamy Elicia da Silva Lopes, João Batista da Silva Junior, Ileane
Gurgel Mendes
Universidade Federal Do Rio Grande Do Norte - UFRN
CORDEL FOLK LITERATURE AS A TOOL FOR
PREVENTING LEISHMANIASIS IN BRAZIL
Marcos Paulo Gomes Pinheiro, Maria de Fátima Freire de Melo
Ximenes, Maria Paula de Melo Pereira Pinheiro, Maria de Lima Alves
Universidade Federal do Rio Grande do Norte
marcospinheiro.pinheiro@bol.com.br
malimalves11@gmail.com
Keywords: Cordel Literature, Leishmaniases, Phlebotomine
sandflies, Education.
Keywords: Phlebotomine sandflies, Educational comic strip
booklet, health education.
Leishmaniases, a set of diseases transmitted by phlebotomine
sandflies, are one of the greatest challenges faced by public health due
to the lack of mechanisms, technology, drugs, vaccines and effective
educational measures to control them. The expansion of Visceral
Leishmaniasis (VL) and American Cutaneous Leishmaniasis (ACL)
in Brazil is associated to the adaptation of the vector to environments
modified by anthropic action and the precarious socioeconomic and
sanitary conditions of the population. In Rio Grande do Norte visceral
and cutaneous leishmaniases are expanding. A number of factors
contribute to this situation, in addition to aspects related to the species
and abundance of sandflies in this region. In the year 2012 were
peformed systematic captures of sandflies with CDC light traps in two
counties in the state of Rio Grande do Norte, where showed the
occurrence of species incriminated as vectors of LV and LTA. In the
first countie this state, São Gonçalo do Amarante, was found the
presence of Evandromyia lenti (84%), Evandromyia evandroi (8%),
Lutzomyia longipalpis (6%), Sciopemyia sordellii (1%), Evandromyia
walkeri (0,5%) and Nyssomyia intermedia (0,5%). Already in the
countie of Assu was evidenced lower variation of species
predominating Lutzomyia longipalpis (98%), Evandromyia evandroi
(1,62%), Evandromyia lenti (0,11%) and Micropygomyia
trinidadensis (0,03%). In this sense, the knowledge of sandfly species
contributes to the understanding of the cycle of transmitting the
disease being essential to its control. The use of didactic instruments in
schools and the community is an excellent health education tool for
disseminating knowledge and controlling the disease. The aim was to
create a 28-page educational comic strip booklet containing
information on the cycle of the vector, transmission of the parasite, its
life cycle on different hosts and the forms of leishmaniasis
manifestation. The booklet will be distributed in elementary and
secondary schools as well as to other groups during environmental
health education campaigns aimed at providing knowledge and
recognition of sandflies to the population. The entertaining nature of
comic strips contributes to involving the reader through the words and
images of well-known personalities who represent and legitimize the
scientific discourse and to disease prevention, health promotion and
engaging the population, as well as their participation in issues related
to health and quality of life.
Cordel literature (literally “string literature”) is considered a form of
popular art unique to Brazil, published as rhyming verses, produced in
small inexpensively printed booklets, called cordel because they were
originally hung from strings. This type of literature, in addition to
entertaining, informal and pleasurable, can be easily introduced to
social settings. It consists of folk novels and poems that can be easily
inserted into an educational context to popularize science.
Northeastern Brazil has the highest incidence of visceral
leishmaniasis, affecting primarily the more underprivileged social
classes. The study aims at preventing leishmaniasis via cordel
literature, given its acceptance and entertaining aspect, with a focus
on the transmission, symptoms and prevention of leishmaniases. In
this respect, a booklet entitled “ A drinker of blood in the land of the
mandacaru cactus – the fight against Kalazar” was manufactured to be
used in environmental and sanitary education regarding
leishmaniases, with populations residing near three environment
protection areas in the state of Rio Grande do Norte, in Northeastern
Brazil, two in the Caatinga biome in the municipalities of Assú and
Serra Negra do Norte and one in the Atlantic Forest, located in the
municipality of Nísia Floresta. Phlebotomine sandly collections were
conducted in the aforementioned areas, using CDC traps. Vector
species were captured, highlighting the need for educational
measures. Collections indicate the presence of Psychodopygus
wellcomei, a vector species of cutaneous leishmaniasis in the Atlantic
Forest area, and that Lutzomyia longipalpis, a vector of visceral
leishmaniasis, is the most abundant in anthropized environments, as
well as in the preserved portion of the Caatinga. The most collected
species in the Atlantic Forest area were Lutzomyia walkeri (76.6%),
Evandromyia evandroi (16.5%) and P. wellcomei (1.8%), and in the
Caatinga L. longipalpis (98%) and L. evandroi (1.6%). Given the lack
of information on the topic, educational initiatives were carried out in
a school setting. The interest of individuals in reading the booklets can
be readily observed and we can therefore use them to increase
community awareness, since this type of literature is easily accessed,
very inexpensive, in addition to reaching all age groups, and social
and educational levels, making it an easy, low-cost way to
disseminate information on an tissue of great importance to public
health.
N° 26 (1) 2015
S U R V E Y O F A M E R I C A N T E G U M E N TA R Y
L E I S H M A N I A S I S I N T H E N O RT H E A S T O F
ARGENTINA DURING 2011-2014
CANINE VISCERAL LEISHMANIASIS. IMPORTED
CASES IN URUGUAY, 2010-2014
Lozano Alejandra1; Basmadjián Yester2; Vitale Edgardo1; Satragno
Dinora2; Canneva Bruno2; Verger Lorenzo1; Tort Cecilia2; Viera Ana2;
Romero Selva2; Ríos Cristina3 and Lagarmilla Patricia3
María Josefa Felisa Rea, Carlos Edgardo Borda
Centro Nacional de Parasitología y Enfermedades Tropicales (Cenpetrop), Facultad
de Medicina, Universidad Nacional del Nordeste
cenpetrop@hotmail.com
Keywords: American tegumentary leishmaniasis
Argentina
120
northeast
1
Área de Medicina Preventiva y Epidemiología, Facultad de Veterinaria, Universidad
de la República, Montevideo. Uruguay. Alberto Lasplaces 1550. 2Departamento de
Parasitología y Micología Médica. Instituto de Higiene, Facultad de Medicina,
Universidad de la República. Montevideo. Uruguay. Avda. Alfredo Navarro 3051,
Montevideo, Uruguay. 3Área de Salud Pública Veterinaria, Facultad de Veterinaria,
Universidad de la República. Montevideo. Uruguay. yester@higiene.edu.uy
Keywords: Leishmaniasis visceral, Uruguay, dogs
American tegumentary leishmaniasis (ATL) is an endemic disease, in
some parts of Argentina. Corrientes province, located in the Northeast
region, report many leishmaniasis cases with epidemic outbreaks
recorded in three municipalities. One hundred forty seven human
cases of ATL were diagnosed from 1987-2010 in the CENPETROP.
Leishmania (Viannia) guyanensis, Le. (V.) braziliensis and Le. (Le.)
amazonensis have been isolated from human cases.The purpose of
this study was to describe seven new cases of disease from 20102014, and to identify potential sandfly vectors in peridomicile of two
patients. Four patients were men and three women. Age range was
between 28-82 years old. From the province of Corrientes were five
patients(two from Corrientes city and three from the Departments of
Lavalle, San Luis del Palmar and Bella Vista), one from the province
of Formosa(Fontana)and another from the Paraguay
Republic(Pilar).Ulcers were present in four patients and the legs were
the most affected parts of the body.Two patients presented only
mucosal involvement (nasal and oral) and in one patient with two skin
ulcers, mucosal compromise was observed one year later of the
primary lesion.The Montenegro skin test was positive in all patients
and the average size was 9.3mm and IM Glucantime® was
administered. No relapse of infection was recorded in cured patients,
but in two women with a complete clinical cures both suffered
traumatism in the same place of the lesion scar within four or five
years later, thus they relapsed and a new ulcer was formed. A 64 yearold male was erroneously diagnosed of epithelium cancer, thus he
received a transplantation of tissue. Between 9 and 12 months later
this patient had relapse and a new ulcer was formed in the same place
of transplant and with metastasis in the oral mucous. Using light traps
and sticky-paper traps, sand flies were collected in the houses of two
patients from Corrientes and Bella Vista. In Corrientes three species:
Lutzomyia (Nyssomia) nievai, Lu. (Pintomyia) shannoni and Lu.
(migonei) migonei were collected. The first species was dominant
(86%). In Bella Vista 15 Lu. (Ny.) nievai were collected. In both, Lu.
(Ny.) nievai was the most common species and therefore suspected to
play the main role in ATL transmission. Further studies are needed to
verify the role of some species of sand flies as vectors of leishmaniasis
in the area. The occurrence of these cases demonstrates the
persistence endemic of the disease.
Visceral Leishmaniasis (VL) is a parasitic zoonotic disease caused by
the protozoan Leishmania infantum and transmitted by the vector
Lutzomyia longipalpis. The disease affects several species of
mammals and the dog is considered its main reservoir. In the year
2000 took place the emergence of this disease in the Southern Cone of
the Americas, being the urban cycle the main form of transmission. In
this cycle, the dog plays a major role since, apart from being the
parasite´s main host, is in a close relation with the human being. At the
present date, VL is an exotic disease in Uruguay, since no
autochthonous cases were described in humans or dogs. L.
longipalpis was found for the first time in Uruguay in the year 2010,
in the northern cities of Salto and Bella Unión. Since then, the whole
country was classified as vulnerable for the emergence and
dissemination of VL. The current situation is worsened by the
occurrence of human and canine cases in border locations and the
entry of infected dogs from endemic areas. These dogs enter the
country accompanying immigrant families or people returning to the
country after spending years abroad. In the present work, a
description of the imported canine cases that entered the country is
performed. Between the years 2010 and 2014 six cases were detected,
five already came with a diagnosis of VL and one was diagnosed in a
veterinary practice shortly after its arrival. Three dogs came from
Europe: two from Spain and one from France. The remaining three
dogs came from Paraguay. Once in the country, the owners of the
infected dogs took up residence in the departments of Maldonado,
Canelones and Montevideo. On the follow up of the cases: two dogs
died from the disease, two were euthanized and one (whose owner
refused euthanasia) is still living in Montevideo. Contact was lost
with the remaining two dogs. Given the current epidemiologic
situation of the country, it is essential to have a clear policy on how to
handle the dogs infected. It is also strongly recommended harden the
controls on the dogs coming from areas where VL is endemic and
avoid that any infected dog reach an area where L. longipalpis is
present.
N° 26 (1) 2015
OCCURRENCE OF LUTZOMYIA LONGIPALPIS AND
S PAT I A L D I S T R I B U T I O N O F V I S C E R A L
LEISHMANIASIS IN FLORIANO, PIAUÍ, BRAZIL:
PRELIMINARY DATA
USE OF REMOTE SENSING IN THE ANALYSIS OF
POPULATIONS OF LUTZOMYIA LONGIPALPIS
COMPLEX
Anárya Teresa de Freitas Rocha¹, Simone Melo Silva¹, Maria Regiane
Araújo Soares²; Daniel Costa Fortier²; Diogo Brunno e Silva Barbosa2;
Carlos Henrique Nery Costa2
1
Biologist, Universidade Federal do Piauí; 2Researcher. Universidade Federal do
Piauí.
regianebiologa@yahoo.com.br
Keywords: visceral leishmaniasis, georeferencing, hotspots
Visceral leishmaniasis (VL), caused by Leishmania infantum and
transmitted by Lutzomyia longipalpis, was first recorded in Brazil in
the beginning of the 20th century as a rural disease and has expanded to
urban centers since the 1980's. This work describes the occurrence of
Lu. longipalpis and spatial distribution of human cases of VL in the
municipality of Floriano, Piauí, Brazil, as well as discusses the
influence of environmental variables on their notification. Data on
notification were collected from the Sistema de Informação de
Agravos de Notificação (SINAN) and the Centro de Controle de
Zoonoses (CCZ), and includes the years 2005 and 2007 to 2013. VL
cases were georeferenced using the Global Positioning System,
followed by the preparation of a distribution map (Google Earth
software). Environmental variables (temperature, relative humidity
and rainfall) were obtained by Instituto Nacional de Meteorologia.
Data on sand flies were obtained in 2014, through collections
performed in areas of higher occurrence of VL based on the previous
information provided by SINAN and CCZ. Collections were
quarterly carried out in animal shelters and residences using light
traps. During the study period, 66 cases were reported, with the largest
number of records in 2009 (n=17). Among the districts, “Tiberão”
detained most of the records (n=6), followed by “São Borja” (n=5)
and “Viazul” (n=5), all located in the periurban area of the city. At
“Tiberão”, it was observed that all notifications (n=6) were composed
by two people in the same household in three consecutive years (2008
to 2010). In this district, 496 specimens of Lu. longipalpis (413 ♂ and
83 ♀ ) were collected in the first quarter of 2014, 85% of which in
peridomestic environment. Only 38 cases were georeferenced and
used for the preparation of spatial distribution maps, thus excluding
the cases with incomplete information. A reduction in the number of
cases in the countryside was observed, in contrast to peripheral
districts that represent the expansion area of the city. There was no
correlation (Spearman test) among the environmental variables and
case reporting (temperature, p=0.28, relative humidity, p=0.61, and
rainfall, p=0.69). New technologies are powerful tools for defining
risk areas for action and execution plans to control VL. The
occurrence of Lu. longipalpis in peripheral urban areas suggests that
these areas may provide favorable conditions for the establishment of
natural breeding vector.
121
Mirella FC Santos1, 2; Antonio C Paranhos Filho3; Roberto M Gamarra3;
Wedson D Fernandes4; Reginaldo P Brazil5; Alessandra G Oliveira1, 2, 6
1
Post Graduate Program in Animal Biology, Federal University of Mato Grosso do Sul,
Campo Grande, Brazil. 2Post Graduate Program in Infectious and Parasitic Diseases,
Federal University of Mato Grosso do Sul, Campo Grande, Brazil. 3Geotechnology
Laboratory, Federal University of Mato Grosso do Sul, Campo Grande, Brazil.
4
Federal University of Great Dourados, Mato Grosso do Sul, Brazil. 5Infectious
Diseases Laboratory, Oswaldo Cruz Institute, Oswaldo Cruz Foundation, Rio de
Janeiro, Brazil. 6Parasitology Laboratory, Federal University of Mato Grosso do Sul,
Campo Grande, Brazil.
mirella.santos@ufms.br
Keywords: Vector, leishmaniases, NDVI, NDWI
Visceral Leishmaniasis (VL) is the most severe form of
leishmaniases. In Latin America, the VL etiologic agent is the
protozoa Leishmania (L.) infantum and its main vector Lutzomyia
longipalpis. In Mato Grosso do Sul (MS), there is a second species,
Lutzomyia cruzi, incriminated as a transmitter of the protozoa. In
recent years, there has been a marked development of studies relating
health and environment. Both can be influenced by space position in
where they are inserted, interfering in their conditions and
characteristics. Thus, the objective of this study was to determine
whether environmental factors such as vegetation and the presence of
water, assessed by remote sensing, could explain molecular and
morphometric differences between populations of Lutzomyia
longipalpis complex in MS. For this purpose, the capture points and
the urbanized areas relating to seven populations "longipalpis"
complex were analyzed: a population of Lu. cruzi (Corumbá) five
populations of Lu. longipalpis in MS (Campo Grande Aquidauna
Miranda Tres Lagoas and Bonito) and an outgroup of Lu. longipalpis
from Estrela de Alagoas (AL). Remote sensing was used for
derivation of ecological indices NDVI (Normalized Difference of
Vegetation Index) and NDWI (Normalized Difference of Water
Index). In order to find out if these environmental variables reflect the
molecular and morphometric differences of the populations studied
an ANOVA (F test) was performed through BioEstat 5.0 program. In
all study sites was possible to observe that the sparse vegetation was
predominant which was expected considering that all areas were
urbanized. The higher NDVI values were Estrela de Alagoas and
Corumbá one with a recent urbanization and the other containing
areas with large limitations of occupancy. It is believed that
environmental factors may mask patterns of genetic isolation often
found in studies of population genetics but in this study these biotic
factors in the dimensioned scale would not be exerting selective
pressure. It was observed that the Municipality of Tres Lagoas is the
most urbanized with lower values of NDVI and NDWI. The results of
this study demonstrate that the species Lu. longipalpis is adapted to
the conditions and characteristics of the urban environment as it is
associated with the occurrence of different humidity conditions and
the supply of vegetation.
N° 26 (1) 2015
GEOMETRIC MORPHOMETRY OF NORTHEASTERN
POPULATIONS OF NYSSOMYIA NEIVAI FROM
ARGENTINA
SANDFLIES (DIPTERA: PSYCHODIDAE) OF AGUAS
CALIENTES, LA CONVENCIÓN, CUSCO, PERÚ
a,b
a
c,d
Utgés, María Eugenia ; Fuenzalida Denise ; Parras Matías ; Gould
Ignaciob; Casertano Sergioa ; Salomón, Oscar Daniela, c
a
REDILA Instituto Nacional de Medicina Tropical (INMeT), Ministerio de Salud de la
Nación, Puerto Iguazú, Misiones, Argentina; b Centro Nacional de Diagnóstico e
Investigación en Endemo-epidemias (CeNDIE), ANLIS, Ministerio de Salud de la
Nación, Buenos Aires, Argentina; c Comité Nacional de Investigaciones Científicas y
Técnicas (CONICET), Ministerio de Ciencia, Tecnología e Innovación Productiva,
Argentina; d Instituto de Medicina Regional, Universidad Nacional del Nordeste,
Resistencia, Chaco, Argentina.
122
Rado D1, Pérez JE4, Vilela M3, Quispe D1, Quispe A1, Aguilar EG1, Rangel
E3, Valladares B2, Pacheco R1
1
de Canarias, Universidad de La Laguna, Tenerife, España; 3Laboratório de
Transmissores de Leishmanioses – Instituto Oswaldo Cruz/FIOCRUZ, Brasil;
4
Instituto de Medicina Tropical Alexander von Humboldt, UPCH, Lima, Perú
dirc82@gmail.com
mariaeugeniau@yahoo.com.ar
Keywords: Peru, Sandflies, Cusco
Keywords: vector population, Cutaneous Leishmaniasis, wing
shape, wing size
The province La Convención (Cusco, Perú) is an endemic area of
Cutaneous Leishmaniosis, having the highest prevalence of the
Department of Cusco. Aguas Calientes at 890 m. (Kepashiato Health
Center) reports the highest frequency of cases. The objective of this
study was to determine the sand fly fauna and its populations in Aguas
Calientes. Sandflies were collected using Shannon traps in
extradomiciliary environments and CDC light traps in intra, peri and
extradomiciliary environments. The collections were done during dry
and rainy seasons in 2010-2011. Thirty one species of Lutzomyia and
Brumptomyia genus were identified with 1 new record for Peru:
Lutzomyia sericea, 3 new record for Cusco: L. b. barrettoi, L.
flaviscutellata, L. gantieri; 4 new records for La Convención: L.
octavioi, L. migonei, L. ayrozai, L. guderiani and 2 not described
species: L. (Psychodopygus) sp. (ca. L. (Psychodopygus)
leonidasdeanei) and L. (Helcocyrtomyia) sp. (ca. L.
(Helcocyrtomyia) kirigetiensis). The most predominant species
were L. y. yuilli (47,25%) and L. octavioi (25,97%) with activity peaks
during 21:00-23:00 and 4:00-06:00, respectively. This species were
found in all collecting sites with predominace of L. y. yuilli in
extradomiciliary environments.
Nyssomyia neivai is the principal vector of Leishmania sp. causing
epidemic Cutaneous Leishmaniasis in nine endemic provinces of
Argentina. At the regional scale, studying the heterogenity of
populations can give clues to develop common or differential control
strategies, to study risk potentiality and migratory routes. If
phenotypic variation has environmental and/or genetic causes, its
study could help to detect local populations with potentially important
characters. Geometric morphometry is a relevant tool for quantifying
phenotypic variation and to understand its epidemiological
importance. To assess differences in shape and size between Ny. neivai
populations, we compared wing shape and size between individuals
from three endemic northeastern provinces: Chaco (Puerto
Antequeras, 31 females), Formosa (Las Lomitas, 19 females), and
Misiones (Iguazú National Park, 16 females). We used 12 landmarks
on each wing (image digitalizing: .TPSdig2, Procrustes method and
relative warps calculation: TPSrelw).Centroid size was analized with
a linear mixed model to allow for heterocedastic group variance,
followed by Tukey contrasts (package lme, R software). Mean
centroid size of females from Misiones was found to be greater from
the ones from Formosa (mean (SE): 2285 (21); 2222 (12); p= 0.025).
No evidence of centroid size differences were found between groups
from Misiones and Chaco (p= 0.44), or Chaco and Formosa (p=
0.60).Wing shape was analized by MANOVA and HotellingBonferroni contrasts (Infostat). We found significant differences in
wing shape between Misiones females and Chaco/Formosa groups
(Wilks= 0.41; d.f.: 24, 104; p= 0.0012). Following a discriminant
analysis (Axis 1= 66%), reclassification based on shape showed 71%
of correct validated classification.
N° 26 (1) 2015
124
STUDIES ON FAUNA OF PHLEBOTOMINE SAND
FLIES (DIPTERA: PSYCHODIDAE) AT “TERRA
FIRME” AND “VÁRZEA” ENVIRONMENTS IN TEFÉ
MUNICIPALITY, AMAZONAS STATE, BRAZIL
PHLEBOTOMINE SAND FLY SURVEY IN THE FOCUS
OF HUMAN LEISHMANIASIS IN SOUTH WESTERN
MADRID REGION, SPAIN: 2012-2013
Maribel Jiménez, Estela González, Sonia Hernández, Ricardo Molina
Antonio Marques Pereira Júnior1,2, Eric Fabricio Marialva2, Moreno de
Souza Rodrigues3, Felipe Arley Costa Pessoa3, Jansen Fernandes de
Medeiros4
1
Instituto Nacional de Pesquisas da Amazônia, 2Centro de Pesquisa Leônidas & Maria
Deane, FIOCRUZ AMAZÔNIA, 3Centro de Pesquisa Gonçalo Muniz, FIOCRUZ
BAHIA, 4FIOCRUZ Noroeste
rmolina@isciii.es
Keywords: Phlebotomus perniciosus, Leishmania infantum, sand fly
infections, Spain
Keywords: Ecology, Amazônia, Epidemiology
some female species are vectors of leishmaniasis. The aim of this work
was to verify phlebotomine fauna at “Terra firme” and “Várzea”
environments and ecotopes (forest and peridomiciles) in Tefé
municipality, Amazonas state, Brazil. The collections were performed
using HP light traps during six months: January, February, April,
August, September and October, of 2013. The specimens were
clarified with Potassium Hydroxide (10%) and mounted on slides. A
total of 5.716 individuals were collected and identified 46 species. The
abundance of the males was slightly higher compared to females,
2.868 males (50.19%) and 2.848 females (49, 81%), respectively.
Trichophoromyia ubiquitalis (3,330 – 58.26%) and Nyssomyia
antunesi (661 – 11.26%) were most abundant. At “Terra firme”
environment 42 species were found (5.428 individuals) and Th.
ubiquitalis, Ny. antunesi and Ny. yuilli yuilli were most abundant
species with 3.312, 526 and 259 individuals, respectively. While at
“Várzea” environment 23 species were found (288 individuals) and
Ny. antunesi, Ev. walkeri were most abundant species with 135 and 87
specimens, respectively. The forest ecotopes had high diversity when
compared to peridomiciles in both environments. At terra firme, the
ecotope forest was richer with 4.689 specimens belonging to 39
species, while in peridomicile were found 739 individuals into 27
species. At várzea, the forest also most abundant with 263 individuals
belonging to 21 species, while peridomiciles ecotope were found 25
specimens into 4 species.
Since 2010 it has increased the number of cases of both visceral and
cutaneous human leishmaniasis in south western Madrid region,
Spain. Surprisingly the prevalence of canine leishmaniasis in the area
of the focus is even lower than that detected in the neighbouring areas.
In a preliminary entomological survey carried out in the area in 2011
Phlebotomus perniciosus was the only one potential vector identified.
In order to a better understanding of the new active focus an
entomological survey was carried out from May to October 2012 and
2013 in 4 stations located in a neighbouring green park next to the
urban areas and selected according the higher number of sand flies
collections obtained in 2011 in the same area. Each month 20 sticky
traps (20 x 20 cm) and 2 CDC light traps were used during two
consecutive nights in every station. CDC traps were replaced every
day. The main objective was to establish the phenology of P.
perniciosus in the focus and its relationship to the rates of infection of
this species. A total of 29,729 specimens were collected during both
years, 12,271 sand flies by CDC light traps and 17,458 using sticky
traps. Three sand fly species were identified in 2012: P. perniciosus
(68.57%), Phlebotomus sergenti (0.02%), and Sergentomyia minuta
(31.41%). Data were similar in 2013 with the following species
identified: P. perniciosus (64.35%), Phlebotomus papatasi (0.01%),
and S. minuta (35.64%). Mean densities of P. perniciosus obtained
using sticky traps were 159.01 flies/m2 in 2012 and 146.13 flies/m2 in
2013. On the other hand, 735 P. perniciosus female collected with
CDC traps in the seasonal study of 2012 were dissected and 18 of
them were found infected with L. infantum (2.45%). Infected females
were found in the four stations from July to September. A total of 14
isolates were successfully isolated and further characterized as L.
infantum by PCR of ITS regions. In 2013 a number of 863 P.
perniciosus females were dissected with 57 of them infected, giving a
rate of 6.6%. A total of 44 isolates were obtained in the 4 stations from
July to October and were also characterized as L. infantum. We
present here the evolution of L. infantum infection rates in P.
perniciosus along the transmission season in the four stations studied
during 2012 - 2013. These data provide significant epidemiological
information which is closely linked to the spread of human
leishmaniasis in the focus. This study was funded by EU grant FP72011-261504 EDENext.
N° 26 (1) 2015
LUTZOMYIA SPP. (DIPTERA: PSYCHODIDAE) IN
INTRA, PERI AND
EXTRADOMICILIAR
ENVIRONMENTS IN YOMENTONI (RIGHT MARGIN),
LA CONVENCIÓN, CUSCO, PERÚ
SOME ECOLOGICAL ASPECTS OF LUTZOMYIA
LONGIPALPIS (DIPTERA: PSYCHODIDAE) IN
ENDEMIC AREA OF VISCERAL LEISHMANIASIS,
MATO GROSSO DO SUL, BRAZIL. PRELIMINARY
RESULTS
Toccas F1, Chacon M1, Pérez JE3, Rado D1, Soto M1, Mendoza J1, Luna R1,
Aguilar EG1, Valladares B2, Pacheco R1
1
Tropical Alexander von Humboldt, UPCH, Lima, Perú
flor_13_8@hotmail.com
Keywords: Perú, Cusco, Lutzomyia
Yomentoni at 583 m. (Right Margin) is an endemic area for
leishmaniosis, no studies on Lutzomyia, spp. were done in the past.
The objective of this study was to identify and determine the species of
Lutzomyia in intra, peri and extradomestic. Sand fly collections were
done with CDC light traps and Shannon trap with protected human
attractant during the rainy and dry seasons of 2010 and 2011. Twenty
eight sandfly species were identified, with a new record for the
Province La Convención: L. (P.) trispinosa. Extradomiciliar
environment (44.03%) shown the largest sand fly collections.L.
(Pressatia) sp. was the most frequent species for the three collection
environments and for both the rainy and dry seasons. As well as there
is significant difference (p = 0.000<0.05) for species presence in the
three collection environments in relation with the seasons and sex;
most female collections belongs to L. (Pressatia) sp. and L.
(Trichophoromyia) sp.
Supported by AECID, Spain.
123
Helen Rezende de Figueiredo1, Aline Etelvina Casaril1, Jucelei O. M.
Infran2, Elisa Teruya Oshiro3, Adauto Rodrigues Mendes3, José
Dilermando Andrade Filho5, Letícia Moraes Ribeiro2, Everton Falcão
Oliveira6, Mirella Ferreira da Cunha Santos2, Alessandra Gutierrez de
Oliveira1, 2, 3
1
Post Graduate Program in Infeccious and Parasitic Diseases, Federal University of
Mato Grosso do Sul; 2Post Graduate Program in Animal Biology, Federal University
of Mato Grosso do Sul; 3Parasitology Laboratory, Federal University of Mato Grosso
do Sul; 4Central of Zoonotic Control, Municipality of Aquidauana/MS; 5Center of
National and International Reference in Sandflies, Leishmaniasis Laboratory,
Research Center René Rachou; 6Post Graduate Program in Public Health, São Paulo
University.
mirella.santos@ufms.br
Keywords: Vector, sandfly, seasonality, behaviour
The municipality of Aquidauana, MatoGrosso do Sul, Brazil,
frequently presents cases of canine and human visceral leishmaniasis
and is classified as an area of intense transmission. The sandfly
Lutzomyia longipalpis is the major vector of Leishmania
(Leishmania) infantum, the causative agent of visceral leishmaniasis
in Central and South America. This vector is widely distributed in
both wild and domestic surroundings. This species occurs in areas
with different climatic conditions and presents great adaptability to
different habitats. This study aimed to identify behavioral aspects of
Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae) in the
study area The captures were carried out from April 2012 to March
2014, in six neighborhoods, fortnightly, with light automatic traps
from 06 pm to 07 am, in peri and intradomicile environments each
dwelling. The specimens were transported to the Laboratory of
Parasitology of the Federal University of MatoGrosso do Sul (UFMS)
for mounting, identification, and other observations. During the
capture period 2569 specimens of Lutzomyia longipalpis, 2271 males
(88.4%) and 298 females (11.6%) were collected. Of the total, 71.7%
(1842) were captured in the first year, which showed irregular
distribution over the months. In the second year, the distribution of the
remaining 29.3% (727 individuals) seems to be more regular,
coinciding peaks of higher frequency in wet seasons and peaks of less
frequency in dry seasons. Lu.longipalpis behaved as an annual
species, but was not present in all locations through the month. The
population of the neighborhood 2 (Pinheiro) had the highest
percentage of sand flies (31.9% - 820 individuals), despite not having
been captured in all months. On the other hand, the population of the
neighbourhood 4 (São Cristóvam) had the second highest percentage
of Lutzomyia. longipalpis (28.6% - 737 individuals) and was present
in all months. The sandflies were captured more frequently
peridomiciles 81.08% (2,083) than in households 18.91% (486),
important information, considering the anthropophilic behavior of
this vector. In addition, females were analyzed by polymerase chain
reaction (PCR) in search of Leishmania sp. Analysis was negative,
which was expected, since the natural infection rates, even in endemic
areas, tend to be very low.
N° 26 (1) 2015
MOLECULAR DETECTION OF LEISHMANIA
INFANTUM AND HOST-FEEDING PREFERENCES IN
PHLEBOTOMUS PERNICIOSUS FROM THE FOCUS OF
LEISHMANIASIS IN SOUTH WESTERN MADRID
REGION, SPAIN: 2012-2013
INTERACTION BETWEEN CANINE AND HUMAN
VISCERAL LEISHMANIASES IN A HOLOENDEMIC
FOCUS OF CENTRAL TUNISIA
Maribel Jiménez, Estela González, Sonia Hernández, Ricardo Molina
mjimenez@isciii.es
Keywords: Phlebotomus perniciosus, host feeding preferences,
Leishmania infantum, Spain.
An entomological survey was carried out from May to October 2012
and 2013 in 4 stations placed neighbouring the urban area of the focus
of leishmaniasis in south western Madrid. Direct xenodiagnosis in
hares (Lepus granatensis) and wild rabbits (Oryctolagus cuniculus)
from the focus proved that they are infective to colonized
Phlebotomus perniciosus. Further characterization of promastigotes
isolated from these sand flies fed on both hares and rabbits
demonstrated that these lagomorphs were infected by Leishmania
infantum. The analysis of blood preferences and detection of L.
infantum was studied in a total of 139 blood female sand flies captured
in the transmission period from June to October of 2012 (n=71) and
2013 (n=68). CDC light traps and sticky traps were used placed in four
stations through the entomological surveys. Initially, genomic DNA
was obtained from individual sand flies and used to amplify a 120 bp
fragment of the conserved region of kDNA.. Positive samples were
further analysed following a specific L. infantum PCR based on the
amplification of a fragment of 702 bp of cpb gene. Blood meal
identification was conducted by the amplification of a fragment of 359
bp of vertebrate cytochrome b (cyt b) gene, further sequencing, and
subsequent comparison with sequences deposited in the GenBank.
Data from female sand flies captured in 2012 revealed that blood
meals were mainly rabbits (n=41; 57.8%) followed by hares (n=28;
39.4%). DNA from L. infantum was detected in (n=2; 4.9%) sand flies
fed on rabbits and in (n=11; 39.3%) sand flies fed on hares. Similarly,
blood source from a human and another one from a dog were
respectively found in two sand flies representing 1.43% in each case.
In addition, in the 68 sand flies processed during 2013 blood meal
from rabbits mostly was found in 46 specimens (67.6%), and in 16
(23.5%) blood source from hares was identified. DNA from L.
infantum was detected in ten (21.7%) sand flies fed on rabbits and in
three (18.8%) fed on hares. Also, in five sand flies blood from cats was
found (7.3%) and in one sand fly (1.5%) human blood was identified.
In conclusion, the data revealed an important increase in the number of
sand flies fed on rabbits during the sand fly season, a direct
consequence of the control measures undertaken all over 2012 that
significantly reduced the number of hares in the green park close to the
urban area of the outbreak.This study was funded by EU grant FP72011-261504 EDENext.
125
Z Zoghlami1, E Chouihi1, W Barhoumi1, K Dachraoui1, Nabil Massoudi1,
K Ben Helel2, Z Habboul2, MH Hadhri§, S Limam3, M Mhadhbi3, M
Gharbi3, E Zhioua1
1
Institut Pasteur de Tunis, Laboratory of Vector Ecology, 13 Place Pasteur BP 74, 1002
Tunis, Tunisia. 2Paediatric Department, University Hospital of Kairouan, Tunisia,
§.Regional Department of Health, governorate of Kairouan, Tunisia. 3Veterinary
School of Sidi Thabet, Sidi Thabet, Tunisia
Keywords: Visceral leishmaniasis, Dogs, Humans, Sand fly vectors
Canine visceral leishmaniasis (CanVL) is endemic in the
Mediterranean basin. In Tunisia, CanVL is spatially associated with
human visceral leishmaniasis (HVL) affecting mostly children
younger than 5 years old. In this study, seroprevalence of Leishmania
infantum infection in dogs was assessed in highly endemic districts of
the governorate of Kairouan where more than 50% of HVL cases in
Tunisia were reported. An entomological investigation was also
carried out in two endemic districts (Bouhajla and Haffouz) to assess
sand fly fauna and infection status of sand flies with Leishmania. A
total of 191 serum samples were collected from healthy dogs and
tested for anti-L. infantum antibodies by indirect
immunofluorescence antibody test (IFAT). Overall seroprevalence
was 26.7% being highest among dogs in the district of Bouhajla
(52.7%) and the lowest in the district of Chbika (5.2%). In dogs,
seroprevalence did not differ significantly based on gender or age,
with dogs younger than 1 year showing a higher seroprevalence
compared to older dogs. These findings suggest strong force of
infection in naïve animals in holoendemic regions leading to
emerging high incidence of HVL. Concomitant to the high CanVL
prevalence observed in the Bouhajla district, a significantly high
cumulative HVL incidence also was observed in this district.
Phlebotomus perniciosus and Phlebotomus longicuspis were the
most abundant sand fly species in Bouhajla and Haffouz districts. The
rate of Leishmania-DNA infection in sand flies was 9.4%. This
finding points to spatial correlation between the occurrence of disease
in humans, a high rate of infection in dogs and a high abundance of P.
pernicious and P. longicuspis. Thus, CanVL is the main risk factor for
transmission to humans and subsequently, it is an important
parameter for controlling transmission to humans.
N° 26 (1) 2015
LEISHMANIASIS VECTORS IN THE ALTO PARANÁ
DEPARTMENT, PARAGUAY
STUDIES ON
SAND FLIES (DIPTERA:
PSYCHODIDAE: PHLEBOTOMINAE) OF URBAN
AND WILD AREAS IN THE DISTRICT OF BARRA DO
GUAICUI, VÁRZEA DA PALMA MUNICIPALITY,
Nilsa Gonzalez-Britez1, Nilda Portillo1, Maria Ferreira 1, Martha
Torales2, Nidia Martínez3, Luciano Franco3
126
1
.Dpto. de Medicina Tropical. Instituto de Investigaciones en Ciencias de la Salud,
Universidad Nacional de Asunción. 2 Dpto. de Entomología del Servicio Nacional de
Erradicación de Paludismo SENEPA. 3 Dpto. Programas del SENEPA-Ministerio de
Salud Publica y Bienestar Social
Danyele Franca da Silva1, Cristiani de Castilho Sanguinette1, Gabriel
Barbosa Tonelli1, Aline Tanure1, Rodolfo German Antonelli Vidal
Stumpp2, Felipe Dutra Rêgo1, Célia Maria Ferreira Gontijo1, José
Dilermando Andrade Filho1
nilportillo@gmail.com
1
2
Centro de Pesquisas René Rachou/FIOCRUZ- Belo Horizonte, Minas Gerais.
Universidade Federal de Minas Gerais- Belo Horizonte, Minas Gerais
Keywords: Leishmaniosis, Phlebotominae, Alto Paraná.
danyele.silva@cpqrr.fiocruz.br
Leishmaniasis is a complex diseases caused by protozoa of the genus
Leishmania and are transmitted by the bite of blood-sucking insects,
known in Paraguay under the name of Karachá, belonging to several
genera of the family Psychodidae, subfamily: Phlebotominae. This
work was developed based on the entomological collections made in
several districts of Alto Paraná, with the objective to identify the
species present in the border region between Brazil, Paraguay and
Argentina, and contribute with strategies based on entomological
surveillance. Secondary data was used from the sand flies collected by
the National Service Malaria Eradication using CDC light traps,
containing dry ice, placed 1.5 meters above the ground for two nights
per place. The traps were switched on at 19:00 hrs and the collections
were made the following day at 9:00hrs. The study was carried out
between the periods of June to November 2012. Were selected
collection areas representing different environments favorable for the
vector covering several localities of the Department of Alto Paraná.
Were identified nine species of Plhebotominae, of which three are of
epidemiological importance in the transmission of Leishmania
braziliensis. In order of frequency vectors identified were: Nyssomyia
neivai, collected in San Alberto, Mbaracayú, Hernandarias and Los
Cedrales districts; Migonemyia migonei, in San Alberto, Mbaracayú
and Los Cedrales districts; and finally Nyssomyia whitmani in San
Alberto and Mbaracayú districts. The occurrence of vectors in districts
including rural, semi-urban and urban areas of the border region, is
considered a risk factor for the transmission of cutaneous
leishmaniasis in these areas. Moreover, studies with sandflies are
important to acquire adequate knowledge about the distribution areas
of circulation and population dynamics of the vectors, which is an
important tool for control strategies aimed at reducing the risk of
contact with the vector, as well as the implementation of policies for
the epidemiological control of leishmaniasis.
Keywords: Leishmaniasis, Lutzomyia longipalpis, Nyssomyia
intermedia
: Leishmaniasis are chronic diseases caused by protozoa of the genus
Leishmania. The main mode of transmission of the parasite to humans
and other mammalian hosts occurs through the bite of some infected
female sand flies. The municipality of Várzea da Palma is located in
the north of Minas Gerais State, presenting 202 human cases of
autochthonous cutaneous leishmaniasis (CL) and 37 cases of visceral
leishmaniasis (VL) recorded between 2004 to 2013. Knowing that the
spread of leishmaniasis depends mainly on the presence and
distribution of vectors, the correct identification of the species of sand
flies circulating in endemic areas and knowledge of their ecology can
contribute to the knowledge of leishmaniasis. This study aims to
identify and compare the sand fly fauna in different environments.
The study was conducted in the district of Barra do Guaicui,
belonging to the municipality of Várzea da Palma. From March 2013
to February 2014, monthly, two CDC light traps were installed in the
transition area, six in wild area and two traps in the urban area. A total
of 3,445 sand flies belonging to eight genera and 15 species were
captured. Nyssomyia intermedia (35.99%), Lutzomyia longipalpis
(17.68%), Evandromyia lenti (14.98%), Evandromyia evandroi
(11.12%) were the species with the highest population density. The
urban area accounted for 60.46% of total sand flies collected,
followed by wild area (32.80%) and the transition area (6.73%).
Nyssomyia intermedia, Lutzomyia longipalpis and Nyssomyia neivai
were more abundant in the urban area. Evandromyia lenti and
Evandromyia evandroi had high density in the wild area. The most
representative species in the transition area was Brumptomyia
avellari. Evandromyia sallesi was evenly distributed between urban
and wild areas. Important vector species involved in the transmission
of CT and VL in the area, some of which may have a role in the urban
and wild cycle of leishmaniasis in Barra do Gauicuí area.
Financial Support: PAPES/FIOCRUZ, CNPq, CAPES and
FAPEMIG
N° 26 (1) 2015
E VA L U AT I O N O F T H E A B U N D A N C E O F
PHLEBOTOMINE SAND FLIES IN HOUSEHOLDS
FROM AN AREA OF VISCERAL LEISHMANIASIS
TRANSMISSION IN BELO HORIZONTE, MINAS
GERAIS, BRAZIL
PHLEBOTOMINAE SAND FLY FAUNA IN TWO
URBAN PARKS IN BELO HORIZONTE, MINAS
GERAIS, BRAZIL (DIPTERA: PSYCHODIDAE)
Elisa Neves Vianna1, Maria Helena Franco Morais2, Andréa Sobral de
Almeida3, Paulo Chagastelles Sabroza3, Mariângela Carneiro4 &
Edelberto Santos Dias5
Faculdade de Medicina, Universidade federal de Minas Gerais.
ramarrina@gmail.com
Keywords: receptivity, vulnerability, phlebotomines
Since 1994, Visceral Leishmaniasis (VL) has been suffering a process
of urbanization in Belo Horizonte, with an increase in both canine and
human cases throughout the years. The vector species, Lutzomyia
longipalpis, is present in the entire municipality, both in the peri and
intradomicile. The identification of receptive (presence of organic
material, trees, shady spots, available hosts) and vulnerable
households (presence of attics in the domicile, cracks, broken
windows) with the presence of phlebotomines could aid the actions of
the Ministry of Health. The objective of this study was to validate the
classification of receptive/vulnerable households for the colonization
by phlebotomine sand flies and the risk of VL transmission through an
entomological survey. One-hundred and fifty-three properties located
in neighborhoods from the Northeast Region of Belo Horizonte were
classified into three levels of receptivity/vulnerability (low, medium
and high) by the endemic combat agents. A random sample of 46 of
the 153 properties included in the study was used to obtain data for the
entomological survey. These properties were prospected bimonthly
between May 2012 and June 2013 for three consecutive nights, using
HP traps, one in the peridomicile and another within the domicile.
Among the households classified as high and medium
receptivity/vulnerability, 86% (10) and 77% (25) of them,
respectively, presented infestation by phlebotomines in at least one of
the bimonthly evaluations. The classification levels of the households
did not present an association with the abundance of phlebotomines as
measured through Spearman´s correlation (ρ=-0,7, p >0,05). In total,
752 phlebotomines were captured, 621 in the peridomicile and 131
within the domicile, showing greater captures in January and March.
Although there was no significant correlation among the different
household classification levels and the abundance of phlebotomines,
the households presented an appreciable percentage of infested
houses, making them productive for the insects. The greatest number
of phlebotomines was found in the peridomicile, probably due to the
greater availability of food sources in the environment. The species of
phlebotomine sand flies are being identified and may be related to
environmental and climatic variables, highlighting issues of the
ecoepidemiology of VL in the region and aiding future entomological
survey actions for the Visceral Leishmaniasis Control Program.
127
Paula Cavalcante Lamy Serra e Meira, Bruna Lacerda Abreu, Ana
Paula Lusardo de Almeida Zenóbio, Cristiani de Castilho Sanguinette,
Gustavo Mayr de Lima Carvalho, Lara Saraiva, José Dilermando
Andrade Filho
Keywords: Phlebotominae, Leishmania, cave
Phlebotomine sand flies are often collected in urban areas in Brazil.
This fact is very important, since some species may be involved in the
transmission of Leishmania to mammals. Based on this background,
this study aimed to determine the sand fly fauna in the cave
environment in the Parque Municipal das Mangabeiras and Parque
Paredão Serra do Curral, located in the urban area of Belo Horizonte,
Minas Gerais, Brazil. In order to study the sand fly fauna,
systematized collections were conducted for a year, with monthly
captures from November 2011 to October 2012, using automatic
CDC light traps and non-systematized collections using Shannon
trap. Thus, the sand fly fauna of Parque Municipal das Mangabeiras
presented fourteen species, being the most abundant Evandromyia
edwardsi and Parque Paredão Serra do Curral featured four species,
being the most abundant Sciopemyia microps. In both parks were
collected species that transmit leishmaniasis, such as Lutzomyia
longipalpis, Nyssomyia intermedia and Nyssomyia neivai. Studies on
the fauna and the behavior of sand flies in cave environments are
essential to understand the relationship of these insects with this
ecotope and also assess and monitor areas that may pose risks to the
health of visitors and employees in this type of
environment.Financial Support: FAPEMIG, CNPq.
N° 26 (1) 2015
SPECIES OF PHLEBOTOMINE SAND FLIES
(DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE)
REPORTED IN BRAZILIAN CAVES
ECOLOGY OF SAND FLIES (DIPTERA:
PSYCHODIDAE: PHLEBOTOMINAE) IN THE
NATURAL HERITAGE PRIVATE RESERVE OF SERRA
DO CARAÇA, MINAS GERAIS, BRAZIL
Paula Cavalcante Lamy Serra e Meira and José Dilermando Andrade
Filho
128
Gabriel Barbosa Tonelli, Felipe Dutra Rego, Gustavo Mayr de Lima
Carvalho, Aline Tanure, José Dilermando Andrade Filho
Centro de Pesquisas René Rachou/FioCruz - Belo Horizonte, Minas Gerais
gabrieltonelli@hotmail.com
Keywords: Phlebotominae, cave, Evandromyia
Keywords: Ecotourism; Phlebotominae; Leishmaniasis;
Transmission.
Among the insects that are found in the cave environment, sand flies
are noteworthy, since they are the main vectors of some pathogens of
medical and veterinary importance, among which stand out
Leishmania. Many caves, even before being studied, are open for
visitation and work practices, without any scientific monitoring.
Thus, the loss of geological, biological and ecological information is
immense. In addition, visits to areas where fauna surveys have not
been conducted, especially of the arthropods presents in the locations,
can pose risks to the health of the population who visit and work there.
Based on this background, this study aimed to report the species of
sand flies collected in Brazilian caves. A literature review was
conducted and the species of sand flies, the lithology of these caves
and the state where the registration took place. Until now, 84 species
of phlebotomine belong to 17 genus were reported in Brazilian cave,
of these species, 14 were described from caves. The Evandromyia
genus is the most representative in Brazilian caves with 15 species,
followed by Lutzomyia and Micropygomyia, both with 11 species
caves. Thus, only the species of these three genera, represent 44.05%
of sand flies from caves. Seventeen species reported in Brazilian caves
can be involved in the transmission of Leishmania to mammals. Given
the information above, it is clear that the Brazilian caves are an
important place to collect sand flies, since several species were
described from caves and this ecotypes can host important species are
important in the transmission of Leishmania. Financial support:
Fapemig, CNPq
There are several tropical vector-borne diseases and among them are
leishmaniases. This complex parasitic disease caused by protozoa,
include approximately 21 species of Leishmania, several mammalian
and sand flies species, which act as reservoirs and vectors of
Leishmania. Knowledge of its transmission cycle can provide
understanding of the epidemiology and ecological relationships of
leishmaniasis. The ecotourism is a growing activity in Brazil. The
increase in tourism in the country associated with the medical
importance of sand flies, justify more detailed studies to know the
possible health risks of tourists and workers in these locations. The
Natural Heritage Private Reserve Serra do Caraça (RPPNPC) is
located in the municipalities of Santa Barbara and Catas Altas and
covers an area of 11,233 hectares, with transitional vegetation between
Cerrado and Atlantic Forest. The aim of this study was to study the
sand fly fauna of this site and verify the presence of possible vector
species of Leishmania, to contribute to the control of leishmaniasis
among tourists in RPPNPC. We used 25 CDC light traps distributed in
seven trails in the park. Trails 1 and 2 were exposed in forested areas;
trials 3 and 4 in “campo rupestre” and in one cave; trials 5 and 6 in the
peridomestic areas; and trial 7 in the intradomicile (in the house
provided for researchers) and outer wall of the house. Collections are
being carried out every two months for one year. To date six samplings
were performed, with 342 sand flies collected, distributed in 9 genera
and 17 species. The predominant species were Psychodopygus lloydi
(71.4%) and Pintomyia monticola (6.14%) and the places where there
was a greater number of sand flies were the trail 7 (27.19%), 1 and 6
(24.27%). In recent studies, specimens of Ps. lloydi were found
infected with Leishmania braziliensis in Minas Gerais, which may be
an indication that this species may be involved in the transmission of
Leishmania in this area. Financial support: FAPEMIG e CNPq.
N° 26 (1) 2015
STUDY OF SAND FLIES (DIPTERA: PSYCHODIDAE:
PHLEBOTOMINAE) IN A TRANSMISSION AREA OF
LEISHMANIASIS IN MINAS GERAIS, BRAZIL
ASSESSMENT OF SAND FLIES (DIPTERA:
PSYCHODIDAE) IN AN AREA WITH CANINE
VISCERAL LEISHMANIASIS IN THE MUNICIPALITY
OF PORTO ALEGRE, RIO GRANDE DO SUL, BRAZIL
Aline Tanure, Gabriel Barbosa Tonelli, Felipe Dutra Rêgo, Gustavo
Mayr de Lima Carvalho, Célia Maria Ferreira Gontijo, Gustavo Fontes
Paz, José Dilermando Andrade Filho
Centro de Pesquisas René Rachou/FioCruz- Belo Horizonte, Minas Gerais
linetanure@hotmail.com
Keywords: Phlebotominae, Leishmaniasis
The intense process of urbanization and industrialization cause a
series of changes in urban areas, extending big cities and creating
metropolitan regions. This is the case of the metropolitan region of
Belo Horizonte, capital of Minas Gerais State, which consists of 16
municipalities, including Brumadinho. In recent years Brumadinho
has undergone considerable socio-environmental changes. Casa
Branca is a district of Brumadinho which is undergoing intense
transformation process, where the construction of condominiums
attracts people in search of tranquility and quality of life. However, it
is known that changes in the natural environment are important
factors in the emergence and reemergence of infectious and parasitic
diseases, including leishmaniasis. The municipality of Brumadinho
have reported an increase in cases of cutaneous leishmaniasis (CL)
and visceral leishmaniasis (VL) in the last 5 years, the latter
classifying the program as an area of sporadic transmission,
according to the Ministry of Health, but, transmission of the disease
has not yet been elucidated since no studies on the sand fly fauna are
available. This project investigates the sand fly fauna in Casa Branca
area. For this, 18 CDC light traps were used for three consecutive
nights, exposed in the peridomicile of 9 homes. Until now, a total of
5,284 sand flies were collected belonging to 13 species, Nyssomyia
whitmani (88.9%) were the most abundant species, followed by
Lutzomyia longipalpis (5.86%). Both species are involved in the
transmission of CL and VL which reinforces the need to investigate
the sand fly behavior aspects in Casa Branca and understand their role
in the chain of transmission of leishmaniasis and direct preventive and
control measures. Financial Support: FAPEMIG, CNPq.
129
G D Souza1, 2; A Cardoso1; R M J S de Carvalho1
1
Núcleo de Vigilância de Roedores e Vetores, Coordenadoria Geral de Vigilância em
Saúde, Secretaria Municipal de Saúde de Porto Alegre, RS, Brazil; 2Seção de
Reservatórios e Vetores, Divisão de Biologia Médica, Instituto de Pesquisas
Biológicas-Laboratório Central de Saúde Pública, Fundação Estadual de Produção e
Pesquisa em Saúde, Secretaria Estadual de Saúde do Rio Grande do Sul, RS, Brazil.
getuliods@pop.com.br
Keywords: Canine visceral leishmaniasis; sand flies; Porto Alegre;
Brasil
Canine visceral leishmaniasis (CanVL) was confirmed in the state of
Rio Grande do Sul, Brazil, in 2008. Two years later, the disease was
diagnosed in dogs in a periurban area, in the city of Porto Alegre, Rio
Grande do Sul. Thus, the objectives of this study were to survey the
sand flies species present in this region, to determine the
epidemiological important of these species in the transmission of this
disease and to understand the population dynamics of these vectors to
help vector control activities. Sand flies were collected monthly from
February/2012 to January/2013 in the locality of Lageado, with CDC
light traps, in three environments: inside house, in the peridomestic
area (domestic animal shelter) and in the forest. The sampling effort
was to capture two nights per month, with one trap per environment. A
total of 3,397 sand flies were collected in the three studied
environments, consisting of 1,575 males and 1,822 females. The
species identified were Migonemyia migonei (62.88%), Pintomyia
fischeri (22.49%), Nyssomyia neivai (11.89%), Psathyromyia lanei
(1.77%), Brumptomyia sp. (0.77%), Pintomyia monticola (0.18%)
and Lutzomyia gaminarai (0.03%). Migonemyia migonei showed the
highest relative frequency of occurrence, representing 58.2% of
specimens captured inside house, 65.64% in the peridomestic area
and 61.61 % in the forest, followed by Pi. fischeri (22.58%, 23.66%
and 19.27%, respectively) and Ny. neivai (16.63%, 9.06% and
13.28%, respectively). The largest number of sand flies (n=1,822)
was collected in the peridomicile and in the forest the highest
diversity was found: seven species. Regarding the annual
distribution, almost all sand flies (95.14%) were collected from
November to May, with a large peak of specimens collected in
February. The three main species were found in all months of the year,
except for Pi. fischeri in July. The predominance of Mg. migonei
suggests this species as the main suspect to transmit visceral
leishmaniasis to dogs in the region. Pi. fischeri and Ny. neivai are
known important vectors in the transmission of cutaneous
leishmaniasis in Porto Alegre and they can also act as secondary
vectors for CanVL. For prevention and control, environmental
management should be intensified on the spring and summer. If
necessary, chemical control should be realized in October, with
reapplication of insecticide in the following January. Supported by:
Municipal government of Porto Alegre.
N° 26 (1) 2015
PERSISTENCE OF LUTZOMYIA LONGIPALPIS,
VECTOR OF VISCERAL LEISHMANIASIS, IN THE
CITY OF SALTO, URUGUAY
SEROLOGICAL SURVEILLANCE IN DOGS OF THE
CITY OF BELLA UNIÓN, A RISK AREA FOR
VISCERAL LEISHMANIASIS
Yester Basmadjián1, Bruno Canneva1,
Lorenzo Verger2, Alejandra
2
1
Lozano , Dinora Satragno , Eduardo Supparo2,3, Cirino Sequeira4, Selva
Romero1, Ana Viera1, Cecilia Tort1, Cristina Ríos 5, Patricia Lagarmilla5
and Edgardo Vitale 2
Verger L1; Lozano A1; Vitale E1; Satragno D2; Sequeira C3; Canneva B2;
Basmadjián Y2; Rios C4; Lagarmilla P4; Tort C2; Viera A2; Romero S2
1
Departamento de Parasitología y Micología Médica. Instituto de Higiene-Facultad de
Medicina-UdelaR. Montevideo. Uruguay. Avda. Alfredo Navarro 3051.
yester@higiene.edu.uy; 2Área de Medicina Preventiva y Epidemiología, Facultad de
Veterinaria, UdelaR. Montevideo. Uruguay; 3Liberal Veterinary Professional, Bella
Unión, Departamento de Artigas, Uruguay; 4Área de Salud Pública Veterinaria,
Facultad de Veterinaria, UdelaR. Montevideo. Uruguay5Área de Salud Pública
Veterinaria, Facultad de Veterinaria, UdelaR. Montevideo. Uruguay.
130
1
Área de Medicina Preventiva y Epidemiología, Facultad de Veterinaria, Universidad
de la República, Montevideo. Uruguay. Alberto Las places 1550. 2Departamento de
Parasitología y Micología Médica. Instituto de Higiene, Facultad de Medicina,
Universidad de la República. Montevideo. Uruguay. 3Liberal Veterinary Professional,
Bella Unión, Departamento de Artigas, Uruguay. 4Área de Salud Pública Veterinaria,
Facultad de Veterinaria, Universidad de la República. Montevideo. Uruguay.
vergerlorenzo@gmail.com
yester@higiene.edu.uy
Keywords: Uruguay, dogs, Leishmaniasis
Keywords: Lutzomyia longipalpis, Uruguay, Salto
The city of Bella Union is located in the department of Artigas, in the
northern part of Uruguay. In the year 2010, in this city, two specimens
of L. longipalpis were found for the first time in the country. Further
research has shown several months of continuous presence of the
vector since the year 2013. These findings confirm the city as a risk
area for an outbreak of Visceral Leishmaniasis (VL), an exotic
disease in Uruguay. In the present work, we made a serological survey
of canines using a rapid diagnostic kit for VL (DPP® Leishmaniose
Visceral Canina, BioManguinhos). The sampling was aimed at dogs
showing clinical signs of the disease (skin lesions, cachexy, fever)
and at those who lived inside a 200 meters radius from a site where
specimen of L. longipalpis were caught with a CDC light trap. 200
dogs where tested for VL: 170 living near sites infested with L.
longipalpis and 30 with compatible symptoms. All the test results
were negative. Despite not having been able to obtain a positive result
to the test, further serological samplings are projected in order to keep
the epidemiological surveillance of the risk areas.
Leishmaniasis emerged in the Southern Cone in its human visceral
form in the year 2000, in the Paraguay-Argentina border. Since then,
the rapid expansion to the south of its vector, Lutzomyia longipalpis,
has determined the appearance of human cases in Argentina in the
year 2006 and the presence of infected dogs in the Argentinean city of
Montecaseros, near the border with Uruguay, in the year 2009. With
this background, in February of 2010, an investigation was conducted
in order to determine the presence of the vector in two cities of the
northern part of Uruguay; Salto (capital city of the department with
the same name) and Bella Unión (department of Artigas), catching
specimen of L. longipalpis in both cities. These findings determined
that both departments were classified as “vulnerable” to the
emergence of Visceral Leishmaniasis. After this punctual vector
surveillance, no further research was made. In the present work an
entomologic survey was conducted to determine if the vector was still
present in the city of Salto. In February of 2014, two CDC light traps
were placed two nights in a row in the city Zoo, the same location that
resulted positive in the year 2010 (S 31°23'51.19" W 57°57'51.65").
The insects caught in the traps were identified under
stereomicroscope and the Phlebotominae sand flies were mounted
with Canada balsam for species identification. Tree specimens of L.
longipalpis were identified: two males and one female. These results
confirm that the presence of the vector in the year 2010 was not an
isolated event and rather suggest a continuous presence in the area.
The status of the department as vulnerable for the entry and
dissemination of the causative agent of Visceral Leishmaniasis is
reafirmed. The city of Salto is still the southernmost location were L.
longipalpis was found in Uruguay.
This project was finnanced by the program I+D 2012 of the Comisión
Sectorial de Investigación Científica, Universidad de la República.
This project was finnanced by the program I+D 2012 of the Comisión
Sectorial de Investigación Científica, Universidad de la República.
N° 26 (1) 2015
IDENTIFICATION OF BLOOD MEAL IN LEISHMANIA
INFANTUM VECTORS FROM THE NORTHEAST OF
SPAIN: PHLEBOTOMUS ARIASI AND P. PERNICIOSUS
BY POLYMERASE CHAIN REACTION BASED AND
SEQUENCING METHODS
PHLEBOTOMINE SAND FLIES (Diptera: Psychodidae)
IN A PERI-URBAN SETTING IN THE
METROPOLITAN REGION OF NATAL BRAZIL
Ballart C1, 2; González E3; Ravel C4; Jiménez M3; Abras A1, 2; Molina R3;
Portús M1; Gállego M1, 2
131
Maria de Lima Alves, Maria de Fátima Freire de Melo Ximenes, Marcos
Paulo Gomes Pinheiro, José Hilário Tavares da Silva, Tamy Elicia da
Silva Lopes, Marcel Miranda de Medeiros Silva
Universidade Federal do Rio Grande do Norte – UFRN
-1
Laboratori de Parasitologia, Facultat de Farmàcia, Universitat de Barcelona (UB),
Spain; 2Centre de Recerca en Salut Internacional de Barcelona (CRESIB), Hospital
Clínic-UB, Spain; 3Unidad de Entomología Médica, Servicio de Parasitología, Centro
Nacional de Microbiología, Instituto de Salud Carlos III, Spain; 4Université of
Montpellier, UMR5290 MiVEGEC et Centre National de Référence des
Leishmanioses, Montpellier, France
malimalves11@gmail.com
Keywords: Phlebotomine sand flies, climatic variables, peri-urban
area
mgallego@ub.edu
Keywords: Leishmaniosis, P. ariasi, P. pernciosus, blood meal
In Spain, human leishmaniosis is present as a hypoendemic disease.
The epidemiological cycle is considered to be zoonotic, with dogs
acting as the main reservoir. However, other mammals have also been
implicated as reservoirs for Leishmania infantum, the only
autochthonous species in the country. Two sand fly species belonging
to the genus Phlebotomus subgenus Larroussius are proven vectors,
P. ariasi and P. perniciosus. In some foci the species do not overlap,
whereas in other they are found in sympatric conditions sharing the
vectorial role. While awareness of the disease dates back to the
beginning of the 20th century, it has little history in northern Spain,
where new endemic areas have been found. Since the vectorial
transmission of Leishmania depends on sand fly feeding behavior, the
analysis of their blood meals is of epidemiological interest. Sand flies
where captured with light traps (CDC-like) in northeast Spain
(provinces of Lleida and Girona, Catalonia). Traps were set for one
night in 20 localities near farms or animal stables and kennels. In total,
165 blood-fed sand fly females were analyzed: 121 P. ariasi, 41 P.
perniciosus, and 3 unidentified. The assays were based on specific
amplification and sequencing of the blood meal–derived single copy
prepronociceptin (PNOC) gene or the vertebrate cytochrome b (cyt b)
gene. Mixed blood meals were analyzed by PCR-RFLP. A Genbank
search was carried out for sequence homology. Blood meals were
identified in 39 flies, 25 P. ariasi, 11 P. perniciosus and 3 unidentified.
Among these, blood meals from the following species were
identified: Homo sapiens (3), Canis familiaris (2), Bos taurus (10),
Ovis aries (15), Capra hircus (3), Equs caballus (3), Sus scofra (3),
Oryctolagus cuniculus (1), Mus musculus (1), Gallus gallus (1).
Mixed meals were identified in 4 cases (1 H. sapiens/O. aries, 3
Capra hircus/Ovis aries). The results related to the sand fly species
and sample origin (location of traps and animals present) are
discussed. Financial support: Spanish Ministerio de Ciencia e
Innovación (CG12010-22368-C02-01).
Phlebotomine sand flies are distributed in nearly all faunal regions
worldwide as pathogenic vectors. In Brazil there are a large number of
species. Northeast Brazil has the greatest occurrence of human cases,
either separately or concomitantly of Visceral Leishmaniasis (VL)
and American Cutaneous Leishmaniasis (ACL). The aim was to
identify species of phlebotomine sand flies in a peri-urban area of the
municipality of São Gonçalo do Amarante, Rio Grande do Norte state,
where cases of ACL and VL have been notified. Captures were made
monthly from January 2012 to December 2013 with CDC light traps,
over 12 consecutive hours from 5:30 in the evening to 5:30 in the
morning. A total of 272 specimens were captured in 2012 and 377 in
2013, 63% male and 37% female. Six species were identified:
Evandromyia lenti 84%, Evandromyia evandroi 8%, Lutzomyia
longipalpis 6%, Sciopemyia sordellii 1%, Evandromyia walkeri 0.5%
and Nyssomyia intermedia 0.5%. The results show that Evandromyia
lenti was more abundant, captured in every month of the year and in
the three environments: shrubland 7.5%, intra-domicile 8.5% and
peri-domicile 84%. Temperature and relative humidity apparently did
not interfere in phlebotomine density in the study period. Greater
species diversity occurred in 2012, with six species. Although 2012
and 2013 are considered drought years in the Northeast, a significant
increase in Evandromyia lenti density was observed in March, April
and May, coinciding with the low rainfall of this period. Considering
the urban growth in the metropolitan region of this municipality,
where human cases of visceral and cutaneous leishmaniasis have been
recorded in recent years and E. lenti and L. longipalpis captured in the
peri-domicile and intra-domicile, entomological and epidemiological
surveillance are essential and must be associated to educational
actions in public and environmental health in this municipality.
N° 26 (1) 2015
SANDFLIES (DIPTERA, PSYCHODIDAE,
P H L E B O TO M I N A E ) I N P I TO C O C AV E , A N
ARCHEOLOGICAL SITE IN NORTHERN MATO
GROSSO DO SUL STATE, BRAZIL
FIRST REPORT OF LUTZOMYIA LONGIPALPIS IN
URUÇUCA, COCOA ZONE OF THE SOUTHERN
STATE BAHIA, BRAZIL
Eunice Aparecida Bianchi Galati1, Paulo Silva de Almeida2-3, Marcia
Bicudo de Paula1, Pedro Catarino Costa Filho4, Romoaldo Martins de
Almeida4
1
Universidade de São Paulo (USP), São Paulo, Brazil. 2Universidade Federal da
Grande Dourados (UFGD), Mato Grosso do Sul, Brazil. 3Laboratório Regional de
Entomologia, Dourados, Mato Grosso do Sul, Brazil. 4Coordenação de Controle de
Endemias, Alcinópolis, Mato Grosso do Sul, Brazil
egalati@usp.br
Keywords: Cave, Fauna, Phlebotomine, vectors.
Caves are ecosystems with more constant climatic conditions than
those in the external area. Several species of sandflies have their
breeding sites in caves, others use them as natural shelters and still
others may visit them occasionally. Recently, Alcinópolis
municipality in north Mato Grosso do Sul state (MS) has awakened
great tourist interest due to the presence of several archaeological
sites, including Pitoco cave containing petroglyphs and traces of the
ancient inhabitants (of 11,000 years ago) in its proximity. A project is
underway to characterize the sandfly fauna of Pitoco cave and its
external environment. Monthly captures are being made with CDC
light traps installed inside, at the entrance to and in the external
environment of the cave in the foothills of the Serra do Bom Sucesso.
The data relating to March to May 2014 are here presented and
analyzed in the light of other MS cave ecosystems. A total of 1,641
sandflies (49% females and 51% males) of 19 species have been
captured, 83.2%, 10.5% and 6.3%, respectively, of them in the
interior, at the entrance to and in the external area. Species of four
subtribes were found: BRUMPTOMYIINA (4.6%) of 2 species:
Brumptomyia avellari and Br. pintoi; SERGENTOMYIINA (14.1%)
of 4 species: Deanemyia sp., Micropygomyia acanthopharynx, Mi.
quinquefer and Mi. vonatzingeni; LUTZOMYIINA (57.4%) of 6
species: Evandromyia lenti, Ev. saulensis, Lutzomyia cruzi, Lu.
longipalpis, Pintomyia christenseni and Sciopemyia sordellii and
PSYCHODOPYGINA (23.9%) of 7 species: Martinsmyia oliveirai,
Nyssomyia whitmani, Psathyromyia aragaoi, Pa. bigeniculata, Pa.
brasiliensis, Pa. campograndensis and Psychodopygus davisi. Lu.
dispar, Mt. oliveirai and Mi. quinquefer have been the dominant
species (46.4%, 19.7% and 12.7%, respectively). The species of the
Longipalpis complex: Lu. cruzi and Lu. longipalpis, visceral
leishmaniasis vectors, account for 2.5% of the specimens). Two
undescribed females of Deanemyia have been found, this being the
first register of this genus in MS. Lu. dispar is a species closely related
to Lu. forattinii and Lu. almerioi, both anthropophilic and suspected
as vectors of visceral leishmaniasis agent, which are found in other
cave ecosystems of MS. So in view of the high density of Lu. dispar in
Pitoco cave, its anthropophily, hourly rhythm and potentiality as
Leishmania vector call for further investigation in view of the
touristic interest of the cave.
132
Sílvia Maria Santos Carvalho, Vanessa Cristina Fitipaldi Veloso
Guimarães, Paulo Raimundo Barbosa dos Santos
Centro de Pesquisas Aggeu Magalhães, Fiocruz-PE, Brasil
vanessa@cpqam.fiocruz.br
Keywords: sand flies, Phlebotomine, Lutzomyia longipalpis,
Uruçuca
Visceral leishmaniasis (VL) is a disease caused by Leishmamia
(Leishmania) infantum, which is primarily transmitted by Lutzomyia
longipalpis. In Brazil, this species has a wide geographical
distribution that coincides with outbreaks of disease, has eclectic and
anthropophilic feeding habits and the ability to occupy different
ecological niches. Captures were carried out on a farm in Uruçuca
municipality, Bahia state, during four consecutive nights of each
month using captor of Castro and Center for Disease Control and
Prevention light traps from July/2011 to December/2012 distributed
in intradomicile, peridomicile (chicken house) and forest remnants
from 6:00pm to 6:00am. A total of 151 phlebotomine sand flies
belonging to 7 species were captured including 6 specimens of L.
longipalpis, one male and four females were found in peridomestic
environments and one female in forest remnants. This the first report
of L. longipalpis in this region of Bahia state, a traditionally endemic
area to American cutaneous leishmaniasis. Although captured
sporadically, this finding contributes to knowledge of the
geographical distribution of phlebotomine sand flies and highlights
the need of monitoring of the area due the risk of emergence of cases
VL.
N° 26 (1) 2015
THE SAND FLY FAUNA (DIPTERA: PSYCHODIDAE)
IN ALTO IVOCHOTE, LA CONVENCIÓN, CUSCO,
PERÚ
STUDY OF PHLEBOTOMINES IN TWO WILD AREAS
OF NORTHEASTERN ARGENTINA AND THE
POTENTIAL ASSOCIATION WITH HOWLER
MONKEYS (ALOUATTA CARAYA)
F Toccas1, D Rado1, Anita Quispe1, C Quispe1, B Galindo1, M Chacon1, MA
Quispe-Ricalde2, B Valladares2, R Pacheco1
1
Tropical Alexander von Humboldt, UPCH, Lima, Perú.
flor_13_8@hotmail.com
133
Mariela F Martínez1,2,4; María S Santini2; María Eugenia Utgés1,2; Martín
Kowalewski3,4; Oscar D Salomón1, 4
1
Instituto Nacional de Medicina Tropical, Ministerio de Salud de la Nación, Puerto
Iguazú, Misiones, Argentina-REDILA; 2Centro Nacional de Diagnóstico e
Investigación en Endemoepidemias, Ministerio de Salud de la Nación, Buenos Aires,
Argentina-REDILA; 3Estación Biológica de Corrientes, Museo Argentino de Ciencias
Naturales “Bernardino Rivadavia”, Corrientes, Argentina; 4Consejo Nacional de
Investigaciones Científicas y Técnicas (CONICET), Argentina
Keywords: Sand Fly Fauna, Alto Ivochote, Cusco
marielafmartinez@gmail.com
Alto Ivochote, at 880 m, is an endemic area of leishmaniosis, which
cause an important health problem for the people in this province. The
objective of this research was to identify and study the sand fly fauna
in Alto Ivochote. Sand flies were collected using Shannon traps in
extra domestic environments and CDC light traps in intra, peri and
extra domestic environments during dry and rainy seasons in 2010 –
2011. Thirty six species of Lutzomyia and Brumptomyia genus were
identified. The most frequently species were L. (Nyssomyia) y. yuilli
(32.11%), with predominance in extra-domestic environments, and L.
(Trichophoromyia) sp. (26.15%) with predominance in intra and
peridomestic environments. Activity peaks during 22:00 – 23:00 for L.
(N.) y. yuilli and during 04:00 – 05:00 for L. (T.) sp. were observed.
There is a statistically significant difference in relation to temperature
and presence of L. (T.) sp. (p = 0.021< 0.05) from collections using
CDC traps and in relation to humidity and presence of L.
(Psychodopygus) h. hirsuta (p = 0.048< 0.05) and L. serrana (p =
0.049< 0.05) from collections using Shannon traps.
Keywords: Wild environment; Phlebotomine; Alouatta caraya;
vertical strata
Support by AECID, Spain.
The interaction among the insect vector, the parasite, and the animal
reservoir determines the dynamic of infection of vector borne
diseases. The animal reservoirs of each Leishmania species vary
according to the environment and geographic location, and may
include both domestic and wild animal hosts. Sandflies are found in
different microenvironments of heterogeneous forest ecosystems.
The goal of this study is to describe and compare the Phlebotominae
species composition among two forest strata in Alouatta caraya
sleeping trees and two wild areas inhabited by this primate. The study
sites were Isla Brasilera (IB) Chaco (27°26' S, 59°40' W) and San
Cayetano (SC) Corrientes (30°47' S, 55°40' W). The former is a small
island in the Paraná River characterized by flooded forest with little to
no human contact, and the latter corresponds to the surroundings of a
rural locality exhibiting forest patches surrounded by grasslands. In
both sites A. caraya (black and gold howler monkey) is the most
important arboreal mammal in terms of biomass. Sandflies were
captured with light traps hung close to monkey sleeping trees at 1.5 m
('low') and 6-8 m ('canopy') above the ground and set on for two
consecutive nights. A total of 964 sandflies were collected in IB (n=7
sleeping trees) belonging to 6 species: Nyssomyia neivai (84.6%),
Brumptomyia sp. (12.6%), Migonemyia migonei (2%), Psathyromyia
shannoni (0.6%), Ny. whitmani (0.1%), and Evandromyia corteleziisallesi (0.1%). A total of 1125 sandflies were collected in SC (n=9)
belonging to 7 species: Ny. neivai (50%), Brumptomyia sp. (25.5%),
Mg. migonei (22.8%), Ny. whitmani (0.9%), Ps. shannoni (0.4%), Ev.
cortelezii-sallesi (0.3%), and Pintomyia pessoai (0.1%). Ny neivai
was the most abundant species in both sites. The comparison of Ny.
neivai abundances among sites and heights by nested ANOVA
showed significantly higher abundances in the lower stratum
(F1,12=6.26; p=0.028), but no significant effect of site factor
(F1,12=0.03; p=0.872) or interaction between site and stratum
(F1,12=0.31; p=0.588). The presence of potential vectors of
Leishmania spp. in the canopy of sleeping trees, though less abundant
than in the lower stratum, suggests that A. caraya might serve as wild
mammal blood source, and therefore, as an amplifier of the vector
population or even a potential reservoir host of leishmanias in these
wild areas.
N° 26 (1) 2015
PHLEBOTOMINE SANDFLIES (DIPTERA:
PSYCHODIDAE) IN A PRESERVED AREA OF THE
CAATINGA BIOME IN RIO GRANDE DO NORTE
STATE, BRAZIL
LUTZOMYIA LONGIPALPIS IN BELLA UNIÓN,
DEPARTMENT OF ARTIGAS, URUGUAY. ONE YEAR
OF SURVEILLANCE: JULY 2013- JUNE 2014.
Maria de Fátima Freire de Melo Ximenes; Renata Antonaci Gama,
Tamy Elicia da Silva Lopes, Renato Cesar Melo Freire, Maria de Lima
Alves, Marcos Paulo Gomes Pinheiro, José Hilário Tavares da Silva,
Marcel Miranda de Medeiros Silva
ximenes@cb.ufrn.br
Keywords: Phlebotomine sandflies, Caatinga Biome, Lutzomyia
longipalpis, preserved forest
Phlebotomine sandflies are recognized as Leishmania vectors and
differ from other insects of the order Diptera in their morphological
and environmental traits. Leishmaniases are classified as neglected
diseases by the World Health Organization. In Brazil the number of
human cases in their different forms is growing. The Northeast is the
primary region of occurrence of visceral leishmaniasis with 47% of
the cases. In the state of Rio Grande do Norte cases of Visceral and
Cutaneous Leishmaniasis have been reported separately or
concomitantly. The visceral form exhibits a higher incidence,
reaching a larger number of municipalities. In an attempt at
understanding the aspects that contribute to the incidence of disease,
in expansion in urban areas of municipalities in the region, a
bioecological study of phlebotomine fauna was conducted in the
National Forest of Açu (FLONA-Açu), an environment where native
Caatinga vegetation is preserved. Captures were made between
October 2012 and September 2013 with CDC light traps. A total of
6,577 phlebotomine specimens were captured: Lutzomyia
longipalpis (98%), Evandromyia evandroi (1.62%) Evandromyia
lenti (0.11%) and Micropygomyia trinidadensis (0.03%).
Temperature and relative humidity did not significantly interfere in
phlebotomine density in the study period. However, a significant
increase in Lutzomyia longipalpis was observed after the rainy
season. The high density of L. longipalpis in a preserved environment
of the Caatinga demonstrates the importance of constant
entomological and epidemiological surveillance by public health
sectors, and considering the location of the National Forest in the
urban setting of Assú, health education activities are required to help
control leishmaniases in dogs and humans and mainly, preserve the
natural habitat of phlebotomine species in the Caatinga biome of the
National Forest of Açú.
134
1
1
Basmadjián Yester
; Canneva Bruno
; Verger Lorenzo22;
2
3
Vitale Edgardo ; Sequeira Cirino1 ; Lozano Alejandra
;
1
Satragno4 Dinora1, Tort Cecilia
;
Viera
Ana
;
Ríos
Cristina ; Lagarmilla Patricia4 and Romero Selva1
1
Departamento de Parasitología y Micología Médica. Instituto de
Higiene, Facultad de Medicina, Universidad de la República.
Montevideo. Uruguay. Avda. Alfredo Navarro 3051, Montevideo,
Uruguay. 2Área de Medicina Preventiva y Epidemiología, Facultad
de Veterinaria, Universidad de la República, Montevideo. Uruguay.
3
Liberal Veterinary Professional, Bella Unión, Departamento de
Artigas, Uruguay. 4Área de Salud Pública Veterinaria, Facultad de
Veterinaria, Universidad de la República. Montevideo. Uruguay.
yester@higiene.edu.uy
Keywords: Lutzomyia longipalpis, Uruguay, Leishmaniasis
Visceral Leishmaniasis is a protozoal caused zoonosis that is rapidly
expanding in the Southern Cone of the Americas. Even though
Uruguay does not integrate the dispersal area of this disease, sporadic
cases do occur in neighboring countries (Argentina and Brazil)
involving human and canine infections. The city of Bella Unión, with
a population of 12,200 habitants (“Instituto Nacional de Estadística”
Census 2011) is located in the Artigas Department, on the banks of the
Uruguay River, being a mirror town of the argentinian city Monte
Caseros (Corrientes province), where since 2009 there is record of
dog infected with this disease. In 2010 for the first time in the city of
Bella Unión, individuals of Lutzomyia longipalpis, vector of visceral
leishmaniasis, were found. No further research was done. From July
2013, a monthly sampling was started in the city, using CDC light
traps for two consecutive nights per month. The city was divided into
quadrants of 300 meters length, placing the traps in areas that matched
the the ""worst case scenario"" definition (abundance of vegetation
and presence of dogs among other parameters). Sampling was
conducted between July 2013 and June 2014 with a total of 41
Phlebotominae sandflies captured in 6 different quadrants. The
species identification was performed by microscopic observation
after dissecting and mounting the specimens with Canada balsam all
the individuals being identified as Lutzomyia longipalpis. The
conducted research highlights the continued presence of the vector in
7 consecutive months (October 2013 to April 2014) with the
minimum temperatures between 22°C and 14°C. The persistent
finding of L. longipalpis for more than 6 months including lower
temperatures than described before by literature must be taken into
account. Considering these findings we conclude that the presence of
L. longipalpis in the city of Bella Unión has occurred for at least 4
years considering Bella Unión as a vulnerable area. The presence of
the vector in much of the city is a matter of concern showing that L.
longipalpis has the conditions to live and reproduce. This project was
finnanced by the program I+D 2012 of the Comisión Sectorial de
Investigación Científica Universidad de la República.
N° 26 (1) 2015
PHLEBOTOMINE SAND FLY SURVEY IN CENTRAL
AMAZON, AMAZONAS STATE, BRAZIL WITH A NEW
REGISTER OF LUTZOMYIA CALIGATA
PHLEBOTOMINE SAND FLIES IN A MILITARY
TRAINING AREA IN NORTH-EASTERN BRAZIL
1,2
3
2
Oliveira AFJ , Freitas RA , Pessoa, FAC , Aguiar NO
1
Universidade Federal do Amazonas; 2Centro de Pesquisa Leônidas & Maria Deane
ILMD – FIOCRUZ AMAZÔNIA, 3Instituto Nacional de Pesquisas da Amazônia
134
155
Abilene Cristina de Arruda Moura, Kamila Gaudêncio da Silva, Débora
Elienai de Oliveira Miranda, Miriele Ramalho, Fernando José da Silva,
Sinval Pinto Brandão-Filho, Domenico Otranto, Filipe Dantas-Torres
1
Departamento de Imunologia, Centro de Pesquisas Aggeu Magalhães, 50670-420
Recife, Pernambuco, Brazil, Dipartimento di Medicina Veterinaria, Università degli
Studi di Bari, 70010 Valenzano, Bari, Italy.
some species are vectors of leishmaniasis. There are in Brazil around
400 sand flies species registered and more than 120 of then occur in
Amazonas State. The incidence of human cutaneous leishmaniasis
cases nearby Manaus and Presidente Figueiredo municipalities are
high. Collections of phlebotomine sand flies were done in some
forested área of Manaus and Presidente Figueiredo, during different
periods of the years of 1998, 2012 and 2013, using CDC light traps and
manual collections in tree trunks using a CDC trap as an aspirator. We
collected 5.300 individuals, distributed in 12 sub genera, six species
groups and 63 species. The species more frequent were L. anduzei, L.
davisi, L. flaviscutellata, L. hisurta, L. olmeca nociva, L.
squamiventris and L. umbratilis, all of then evolved as a vectors of
different species of Leishmania. We collected L. caligata, in
Presidente Figueiredo and it is the first occurrence of this species to
Amazonas State.
Keywords: Sand flies, ecology and Leishmania
Sand flies (Phlebotominae) are vectors of several viral, bacterial, and
protozoal pathogens, including those of the genus Leishmania, the
causative agents of leishmaniasis. The ecology of sand flies is of
interest because it may provide background information for a better
understanding of the epidemiology of leishmaniasis and to predict the
spatial and temporal variations in the risk of leishmaniasis in a given
area. The objective of this study was to identify the diversity of
species that make up the sand fly fauna in a remnant of Atlantic Forest
located in the Campo de Instrução Marechal Newton Cavalcanti
(CIMNC), Paudalho municipality, Pernambuco state, north-eastern
Brazil. From July 2012 to July 2013, light traps were placed (ca.1.5 m
above the ground) in 10 sampling sites (P1-P10) near the burrows of
wild animals, trunks and roots of large trees in shady and humid
environments. Light traps worked from 18:00 until 6:00 am, for 4
consecutive days each month. The insects captured were initially
examined under a stereomicroscope, separated according to sex and
kept in 70% ethanol. The specimens were mounted on slides and
identified morphologically. A total of 14,403 specimens belonging to
23 species were identified, as follows: Lutzomyia choti(n=10,968), L.
longispina(n=1,482), L. sordellii(n=657), L. complexa(n= 650), L.
walkeri(n=182),L.amazonensis(n=113),L.wellcomei(n=84),L.evand
r o i ( n = 5 4 ) , L . q u i n q u e f e r ( n = 5 2 ) , L .
ayrosai(n=48),L.naftalekatzi(n=23),L. barrettoi barrettoi(n=20),
L.claustrei(n=20), L. umbratilis(n=16), L.schreiberi(n=11),
L.capixaba(n=7), L.brasiliensis(n=4), L.viannamartinsi(n=3), L.
whitmani(n=3), L. shannoni(n=3), L. migonei(n=1), L. yuilli
pajoti(n=1) and L. oswaldoi(n=1). The number of males (n=7,278)
and females (n=7,125) was close to unity. The overall monthly
average number of sand flies captured per trap was 110.8, with peaks
in March (169.1 sand flies per trap) and April 2013 (332.4 sand flies
per trap). This study indicates a high level of sand fly diversity,
revealing the occurrence of eight potential vectors of Leishmania spp.
in the examined area, including L. complexa, L. amazonensis, L.
wellcomei, L. ayrosai, L. umbratilis, L. whitmani, L. shannoni and L.
migonei. It also shows a high abundance of sand flies in the study area,
with peaks after raining periods. These data will help us to predict the
period in which there may be an increased risk of Leishmania
transmission in remnants of Atlantic Forest in north-eastern Brazil.
N° 26 (1) 2015
R E L AT I N G V E C TO R S P E C I E S P O T E N T I A L
DISTRIBUTION AND RICHNESS TO TRANSMISSION
FOCI OF CUTANEOUS LEISHMANIASIS IN
COLOMBIA
RISK OF LEISHMANIOSIS TRANSMISSION IN THE
PYRINEAN REGION OF SPAIN ASSESSED THROUGH
THE ISOENZYMATIC ANALYSIS OF POTENTIAL
SAND FLY VECTORS (PHLEBOTOMUS ARIASI AND P.
PERNICIOSUS)
Cristina Ferro, Marla Lopez, Patricia Fuya, Ligia Lugo, Juan Manuel
Cordovez, Camila Gonzalez
Universidad de los Andes, Instituto Nacional de Salud
135
Ballart C1,2; Pesson B3; Martín-Sánchez J4; Alves-Pires C5; Morillas F4;
Afonso MO5; Portús M1; Gállego M1,2
1
c.gonzalez2592@uniandes.edu.co
Keywords: Phlebotominae, distribution, eco-epidemiology,
Leishmaniasis
Laboratori de Parasitologia, Facultat de Farmàcia, Universitat de Barcelona (UB),
Spain; 2Centre de Recerca en Salud Internacional de Barcelona (CRESIB), Hospital
Cínic-UB, Spain; 3Laboratoire de Parasitologie, Faculté de Pharmacie, Université de
Strasbourg and , France; 4Departamento de Parasitología, Facultad de Farmacia,
Universidad de Granada, Spain; 5Unidade de Parasitologia e Microbiologia Médicas
(UPMM), Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa,
Lisboa, Portugal
mgallego@ub.edu
Leishmaniasis is a complex parasitic disease with worldwide
distribution, caused by various species of the genus Leishmania
related to different clinical manifestations. In America, Leishmania is
transmitted by vector insects of the genus Lutzomyia, who maintain
the enzootic cycle in distinct regions by circulating between sylvatic
and domestic mammals, as well as humans who enter the cycles as
accidental reservoirs. In Colombia, Leishmaniasis is an endemic
disease with 95% of all cases being cutaneous. Cases are reported in
several regions of the country with diverse ecological characteristics,
nevertheless, factors like displacement of human populations from
endemic areas, and the intervention of sylvatic areas by the
introduction of agriculture and other socioeconomic and
environmental factors seem to have an impact on the rearrangement of
new transmission cycles. Our study aimed to analyze the relationship
between vectors' distribution and the number of cases reported of
Cutaneous Leishmaniasis (CL) in Colombia. Nineteen sand fly
species reported as of medical importance for the transmission of CL
in Colombia were included and ecological niche modeling was
performed to explore the potential distribution of vector species in
Colombia and their relation to transmission areas and ecosystems. In
total, 560 single records were obtained with L. gomezi as the species
with the highest number of records and N. ylephiletor y P. carrerai
thula with the lowest. Lutzomyia gomezi and P. panamensis had the
widest distribution in contrast with P. spinicrassa, P. youngi, and P.
nuneztovari which was more restricted to a specific region. Potential
species' richness was calculated and overlapped to the average
reported cases from 2005-2013 in Colombia. The departments with
the highest number of cases reported corresponded to Antioquia,
Nariño, Tolima, Meta, Santander and Bolivar. The results showed that
areas with highest prevalence have 2-4 different vector species
predicted as present. We were able to conclude that distribution of
vectors with medical importance in Colombia correspond to disturbed
areas augmenting the domestication potential; vector species with
similar distribution are affected by the same climatic variables, and
finally, we highlight the importance of the use of distribution maps as a
tool for the development of strategies for prevention and control of
diseases.
Keywords: Phlebotomus ariasi, P. perniciosus, Isoelectrofocusing,
leishmaniosis risk
Leishmaniosis, human and canine, does not have a uniform
distribution in Spain, and some northern regions are not considered
endemic. Nevertheless, in the last years, cases of canine leishmaniosis
have been found in some of these northern areas, for example, in the
Pyrenean region. Leishmaniosis transmission is highly specialized,
with sand flies acting as vectors. In Spain, only the females of two
species, Phlebotomus perniciosus and P. ariasi, are confirmed vectors
of Leishmania infantum.. Besides morphological identification,
several new techniques are being applied in the study of sand flies.
These techniques distinguish between cryptic species and
populations with different vectorial capacity, and are therefore useful
in entomological and epidemiological research on leishmaniosis. The
application of the new technology assists in the risk of leishmaniosis
transmission in new areas. In this work, we applied isoelectric
focusing (IEF) to study sandflies captured in northern Spain. The sand
flies (522 P. ariasi and 37 P. perniciosus) come from two Pyrenean
counties of Lleida province (Catalonia): Pallars Jussà (170
specimens) and Pallars Sobirà (389). An IEF analysis using eight
enzyme loci was carried out (PGI, PGM, HK, FUM, ACO, MDH1,
MDH2 and 6PGD). The results were compared with those of other
sand fly populations of leishmaniosis-endemic areas in Spain, France
and Portugal. In the case of P. ariasi, the specimens were grouped
with populations from southern France (Aston and Vira), while two
other distinct groups were found, one consisting of populations from
Cheires (Alto Douro, Portugal) and Rio Tinto (Huelva, Spain) and the
other group from the French towns of Les Tourrettes (La Drôme
Departement) and Roquedur (Gard Departement). Similar results
were found in the case of P. perniciosus, the Pyrenean populations of
Lleida being grouped with others from a highly leishmaniosisendemic area of southern Catalonia (Priorat, Tarragona) and the east
of France (Drôme), while the population of southwestern Spain
(Huelva) has a greater similarity to the one of Portugal. The results
show that sand fly populations of Lleida province in northern Spain,
where leishmaniosis is poorly studied and was considered non
endemic, resemble those of other areas of Spain and France where the
disease is endemic. Autochthonous cases of canine leishmaniosis
were recently diagnosed in this area. Financial support: Spanish
Ministerio de Ciencia e Innovación (CG12010-22368-C02-01).
N° 26 (1) 2015
IRRIGATION IN THE ARID REGIONS OF TUNISIA
IMPACTS THE ABUNDANCE AND APPARENT
DENSITY OF SAND FLY VECTORS OF LEISHMANIA
INFANTUM
SANDFLIES (DIPTERA: PSYCHODIDAE:
PHLEBOTOMINAE) FROM THE NISIA FLORESTA
NATIONAL FOREST, METROPOLITAN REGION
NATAL, RIO GRANDE DO NORTE STATE, BRAZIL
Walid Barhoumi1, Whitney A. Qualls2, Reginald Archer3, Douglas O
Fuller3, Ifhem Chelbi1, Saifedine Cherni1, Mohamed Derbali1, Kristopher
L Arheart2, Elyes Zhioua1 and John C Beier2
Marcos Paulo Gomes Pinheiro, Maria de Fátima Freire de Melo
Ximenes, Maria de Lima Alves, João Batista da Silva Júnior, José Hilário
Tavares da Silva, Marcel Miranda de Medeiros Silva, Tamy Elicia da
Silva Lopes
1
Laboratory of Vector Ecology, Pasteur Institute of Tunis, 13 Place Pasteur BP 74, 1002
Tunis, Tunisia; 2Department of Public Health Sciences, University of Miami Miller
School of Medicine, Miami, Florida, United States of America; 3Department of
Geography and Regional Studies, University of Miami, Miami, Florida, United States of
America
Keywords: Human visceral leishmaniasis, Phlebotomus perfiliewi,
integrated vector management, remote sensing approaches
The distribution expansion of important human visceral leishmaniasis
(HVL) and sporadic cutaneous leishmaniasis (SCL) vector species,
Phlebotomus perfiliewi and P. perniciosus, throughout central Tunisia
is a major public health concern. This study was designed to
investigate if the expansion of irrigation influences the abundance of
sand fly species potentially involved in the transmission of HVL and
SCL located in arid bioclimatic regions. Geographic and remote
sensing approaches were used to predict the density of visceral
leishmaniasis vectors in Tunisia. Entomological investigations were
performed in the governorate of Sidi Bouzid, located in the arid
bioclimatic region of Tunisia. In 2012, sand flies were collected by
CDC light traps located at 9 irrigated and 9 non-irrigated sites to
determine species abundance. Eight species in two genera were
collected. Among sand flies of the subgenus Larroussius, P. perfiliewi
was the only species collected significantly more in irrigated areas.
Trap data were then used to develop Poisson regression models to map
the apparent density of important sand fly species as a function of
different environmental covariates including climate and vegetation
density. The density of P. perfiliewi is predicted to be moderately high
in the arid regions. These results highlight that the abundance of P.
perfiliewi is associated with the development of irrigated areas and
suggests that the expansion of this species will continue to more arid
areas of the country as irrigation sites continue to be developed in the
region. The continued increase in irrigated areas in the Middle East
and North Africa region is of high importance as it is associated with
the spread of L. infantum vector P. perfiliewi. Integrated vector
management strategies targeting irrigation structures to reduce sand
fly vector populations should be evaluated in light of these findings.
136
marcospinheiro.pinheiro@bol.com.br
Keywords: Phlebotomine sandflies, Leishmaniasis, Atlantic Forest.
Phlebotomine sandflies, important insects in the medical-veterinary
field, are vectors of Leishmania protozoa, which cause visceral and
cutaneous leishmaniasis, serious diseases if left untreated, and widely
distributed in Brazil. The present study aimed at assessing the
composition, abundance and diversity, as well as the relationship with
climatic factors of leishmaniasis-transmitting phlebotomine
sandflies, in the Nisia Floresta National Forest (FLONA). This
FLONA, a conservation and research unit covered by different plants
exotic and native to the Atlantic Forest, is situated in the metropolitan
region of Natal, Rio Grande do Norte state, Northeastern Brazil. Two
monthly captures were made over a one-year period, from May 2012
to April 2013, in CDC light traps placed at four points, two more
anthropized, near the occupied area of FLONA, one in an Atlantic
Forest area, and the other in the Pinus forest located within this area. A
total of 2,999 specimens of ten phlebotomine sandfly species were
captured as follows: Evandromyia walkeri, Evandromyia evandroi,
Psychodopygus wellcomei, Sciopemyia sordellii, Psathyromyia
brasiliensis, Lutzomyia longipalpis, Evandromyia lenti,
Psathyromyia shannoni, Nyssomyia whitmani and Nyssomyia
intermedia. The area with greatest sandfly abundance was the Atlantic
Forest (32.4%), followed by the bamboo plantation (26.3%), and with
greatest diversity in the Pinus forest (H'=1.097), followed by the
Atlantic Forest (H'=1.008). The most abundant species was E.
walkeri, with 76% (SISA=1.0), followed by E. evandroi with 16.5%
(SISA=0.87). It is important to underscore the presence of L.
longipalpis, vector species of L. infantum, with 0.64% (SISA=0.80),
as well as P. wellcomei with 1.8% (SISA=0.82), N. intermedia with
0.03% (SISA=0.04) and N. whitmani with 0.10% (SISA=0.04),
species present in cycles of cutaneous leishmaniasis in different
regions of the country. In general phlebotomine sandflies were more
abundant in the rainy season. L. longipalpis was more adapted to
environments with greater anthropic intervention, since it represented
79% of the sandflies captured, while P. wellcomei was associated to
more preserved areas, with 80% in this type of environment and the
rainy season.
N° 26 (1) 2015
ENTOMOLOGICAL SURVEY OF PHLEBOTOMINE
SAND FLIES (DIPTERA: PSYCHODIDAE) IN TURKEY
DISTRIBUTION OF SANDFLIES SPECIES IN A
LOCALITY ENDEMIC FOR AMERICAN
TEGUMENTARY LEISHMANIASIS FROM THE
NORTH OF ARGENTINA, BORDERING WITH
BOLIVIA
Ozge Erisoz Kasap, Bulent Alten
137
Department of Biology, Hacettepe University, Ankara, Turkey
ozgeerisoz@yahoo.com
Keywords: distribution, sand flies, Turkey
In Turkey, visceral and canine leishmaniasis caused by Leishmania
infantum are endemic along the Aegean and Mediterranean costs
while cutaneous leishmaniasis caused by Leishmania tropica and
Leishmania infantum is endemic in the south and southeast of
Anatolia. Hence, most of the sand fly sampling efforts has been
focused on these regions to determine the possible vector species so
far. However, there is no detailed data of the sand fly fauna of the
regions where no leishmaniasis or other sand fly borne diseases cases
have never been reported. In this work we present the results of an
intensive entomological survey carried out in Turkey. Sand fly
samplings were conducted in 19 provinces covering a wide range of
area including the Black Sea, Thrace, Aegean, Southern, Eastern and
Central regions of Turkey between 2005 and 2013. Sand flies were
found to have a wide geographical and altitudinal distribution ranging
from 36 and 1410 meters above sea level. A total of 3846 specimens
belonging to 12 species of Phlebotomus and two of Sergentomyia
were collected. P. perfiliewi s.l. (23.17 %), P. major s.l (23.01 %), and
P. tobbi (16.23 %) composed the most the sand flies collected. P.
papatasi, P. alexandri, P. sergenti, P. balcanicus, P. halepensis, P.
simici, P. burneyi, P. kandelakii, Transphlebotomus sp., S. dentata and
S. minuta were recorded as the other species found in the sampling
locations. Larroussius species which are the proven or probable
vectors of L. infantum in the Mediterranean region were found to have
the widest distribution. Finding these species in almost all the
sampling locations should be considered as a risk factor for the
spreading of the leishmaniasis in Turkey.
Copa GN 1,3, Marco JD 3,5, Nasser JR 1,2 Gil JF 1,2,3,4
1
Instituto de Investigaciones en Enfermedades Tropicales (IIET), Sede Regional Orán,
Facultad de Ciencias de la Salud, Universidad Nacional de Salta (UNSa). 2Cátedra de
Química Biológica y Biología Molecular, Facultad de Ciencias Naturales, (UNSa).
3
Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET).
4
Instituto de Investigación en Energías No Convencionales, Departamento de Física,
(UNSa). 5Instituto de Patología Experimental, Facultad de Ciencias de la Salud,
(UNSa).
noex02@gmail.com
Keywords: Sandflies, American tegumentary leishmaniasis, Aguas
Blancas, distribution
Some sandflies species can vectorize Leishmania protozoan parasites
which cause America tegumentary leishmaniasis (AtL). This disease
is endemic in Argentina, with high incidence in department of Oran,
Salta Province. Previous studies have reported the presence of sand
flies in environments that include wilderness areas, secondary
vegetation, rural, peri-urban and urban areas. The Aguas Blancas, city
located in Oran, is scenary of a fluid transit of people between
Argentina and Bolivia. The aim of this study was to analyze and
describe the abundance and geographic sandflies distribution in
Aguas Blancas, border locality of the north of Argentina. The study
was conducted in October 2013 in nine sampling sites, which
corresponded to residential courtyards widely distributed in the town.
The capture of sandflies was performed by CDC traps placed three
consecutive nights from 19:00 pm to 7:00 pm. The distribution maps
were generated using ArcGIS 9.2 software and statistical analysis
was performed using the chi-square test. A total of 230 specimens
were captured: cortelezzii complex (30%), Evandromyia sallesi
(28.26%), Nyssomyia neivai (24.35%), Mygonemyia migonei
(16.52%). The 12.87% of the females were in a gravid state. The sites
with higher sandflies abundance were located at the edges of the city,
decreasing to zero in the center (p<0.0001). The cortelezzii complex
was the most abundant and showed a wider geographical distribution
that Ny. neivai (p<0.001). Thus, this is the first report for rain forest
region, in which Ny. neivai there is not found as the dominant specie.
Sandflies species found during this study are suspected of
transmitting AtL. Since Aguas Blancas is an area with constant flow
of persons for a migration process through the customs office located
in the city. Thus, it is important to consider the risk of transmission of
this disease specially when considering the potential exposure of nonimmune population from different places of Argentina and Bolivia.
N° 26 (1) 2015
EFFECTIVENESS OF A RAPID MULTIDISCIPLINARY
P R O TO C O L F O R T H E E P I D E M I O L O G I C A L
E VA L U AT I O N O F A N A C T I V E F O C U S O F
C U TA N E OU S L E I S HMA N I A S I S : R E ME D I OS ,
ANTIOQUIA, COLOMBIA
ABUNDANCE ASSOCIATION OF NYSSOMYIA
WHITMANI WITH THE NUMBER OF FARM ANIMALS
AND ITS DWELLING DISTANCE TO THE HOUSE, IN
AREAS WITH TEGUMENTARY LEISHMANIASIS
TRANSMISSION IN PUERTO IGUAZÚ, MISIONES,
ARGENTINA
Freddy Ruiz-Lopez1, Horacio Cadena1, Andrés Vélez Mira1, Laura
Posada-López 1,2, Angelica Contreras1,2,;Didier Tirado Duarte1, Iván
Darío Vélez1
1
PECET Programa de Estudio y Control de Enfermedades Tropicales, Universidad de
Antioquia, Medellín, Colombia. 2GSM Grupo de Investigación en Sistemática
Molecular, Universidad Nacional sede Medellín, Colombia.
138
Villarquide ML1,2; Fernández MS3,4; Santini MS2; Salomón OD3,4
1
Fundación H.A. Barceló;2 Centro Nacional de Diagnóstico e Investigación de
Endemoepidemias-REDILA;3Consejo Nacional de Investigaciones Científicas y
Técnicas;4Instituto Nacional de Medicina Tropical- REDILA
mlucrecia.v@gmail.com
lauraposada22@gmail.com
Keywords: cutaneous leishmaniasis, epidemiology, Lutzomyia.
Keywords: Tegumentary Leishmaniasis, Leishmania braziliensis,
Nyssomyia whitmani.
Leishmaniasis (L) is a group of infectious diseases transmitted by the
bite of female insects belonging to the genus Phlebotomus and
Lutzomyia. In humans leishmaniasis can affect the skin (cutaneous L),
mucous membranes (mucosal L) and internal organs (visceral L). In
recent decades human migration, unplanned urbanization, mining and
climate change have facilitated the adaptation of insect vectors and
wild mammals to peridomestic habitats causing human epidemic
outbreaks in urban and suburban areas. The aim of this research was to
evaluate the epidemiology of an active focus of cutaneous L in
Colombia by optimizing and applying the protocol described by the
National Institute of Health of Colombia. A rapid protocol was
developed by this study for implementation over a period not
exceeding five days. The methods included: 1 Ecoepidemiological
surveys; 2 Blood sampling of patient; 3 Capture and sampling in
mammals; 4 Application of the Montenegro skin test; 5 Sand fly
collection; 6 Laboratory tests; 7 Identification of Lutzomyia species.
Thirty five homes were visited and blood samples were taken from
eight people with suspicious skin lesions. Of these, seven were
positive by direct smear and the parasite was isolated in six of these
samples. Restriction Fragment Length Polymorphism (RFLP)
identified Leishmania panamensis as the circulating strain in this
region. A total of 75 sand flies were collected that grouped into nine
species including Lu. gomezi, Lu. panamensis, Lu. trapidoi and Lu.
yuilli yuilli, all of which are recognized Leishmania vectors in
Colombia. Five mammals were captured, but none were found to be
infected. The area studied is characterized by unplanned human
settlements located on village outskirts and is surrounded by clusters
of forest, where the main source of work is the extraction of gold in a
mining shaft. People are infected due to the close proximity of their
homes to wooded areas that promote the natural transmission cycle of
leishmaniasis. The presence of Leishmania vectors, wild mammal
intermediate hosts and human settlements is indicative of high risk
environments for cutaneous L transmission. These results show that
short studies of endemic foci, addressing all important components in
the transmission cycle of the disease, might be sufficient for the
epidemiological evaluation of regions with active cases of cutaneous
L. For this a multidisciplinary team is required to address all the
components of the transmission cycle.
This study was performed in Puerto Iguazú, Misiones, Argentina, in
an area of recent deforestation-human settlement called '2000
hectáreas'. Tegumentary leishmaniasis (TL) is recorded there since
the 2004-2005 outbreak. Leishmania braziliensis was the circulating
parasite and Nyssomyia whitmani the incriminated vector. The aim of
this work was to study the association between the abundance of Ny.
whitmani and the amount of breed animals (chickens and pigs), and
the vector abundance related to the distance between the house and
the animal dwellings or the forest edge. To develop this study 17 farms
were selected. In each farm, Phlebotominae abundance was measured
in the house, henhouse, pigsty and forest edge (sites). Captures were
done at each site using mini CDC traps on one night during February
2010 (5 pm to 8 am). Number of chickens and pigs, and distance from
sites to the house were recorded for each farm. Similarity between
house, henhouse and forest edge was calculated with the Sörensen
index. Linear regression models for Ny. whitmani´s abundance were
constructed with environmental characteristics as explanatory
variables. Abundance at all the same sites added of Ny. whitmani per
site was: house, 263; henhouse, 3675; pigsty, 1392; and forest edge,
1546. The highest similarity between sites was seen between
henhouse and forest edge (Is=0.39). The abundance of Ny. whitmani
was found to be positively related with number of chickens and pigs
(R2=0.25; SE=0.03; CI95%= -0.01, 0.13; R2=0.47; SE=0.22;
CI95%= -0.03, 1.02, respectively). Forest edge was positively
associated to number of chickens and negatively with distance to the
house (R2=0.83; SE=0.01; CI95%= -0.003, 0.045; SE=0.006;
CI95%= -0.047, -0.019, respectively). Therefore, in scenarios of
recent deforestation/human settlement in Northeastern Argentina, the
likelihood of human-Ny whitmani contact increases with the breeding
of pigs or chickens, that attracts vectors from sand fly source
populations located in the forest or the domestic area. Further, the risk
of transmission is related to the abundance of animals, and the spatial
distribution of the animal dwellings, human sleeping areas and the
edge of the forest. These results suggest that the strategies to prevent
TL in the study area, in human settlements close to the edge of the
forest, should take into account the practices associated with animal
management, and the habits related to the use of the domestic space
(peridomestic and domestic).
N° 26 (1) 2015
BIODIVERSITY OF PHLEBOTOMINAE SUBFAMILY
(DIPTERA: PSYCHODIDAE) IN THE COLOMBIAN
AMAZON AND ORINOCO
S A N D F LY V E C T O R S O F C U TA N E O U S
LEISHMANIASIS ON THE WESTERN SLOPES OF
THE ECUADORIAN ANDES
Cristina Ferro, Patricia Fuya, Ligia Lugo, Camila González
Sandra Enriquez1, Jazzmin Arrivillaga1,5, Franklin Vaca1, Washington
Benitez Ortiz1, Oscar Kirstein2, Ibrahim Abassi2, Manuel Calvopiña3,
Katherine Pozo1, Vanessa Romero1, Orlando Chiluisa1, Ernesto
Villacrés1 and Alon Warburg 1,2,4
Instituto Nacional de Salud (Laboratorio de Entomología-LNR), Universidad de Los
Andes (CIMPAT)
crisferrovela@gmail.com
Keywords: Phlebotominae, Biodiversity, Amazonia and Orinoquia
species
The Phlebotominae subfamily, with more than 800 species, known for
its biodiversity and the role played by some of its members as disease
vectors, is widely distributed in our planet. In the New World
Phlebotominae specimens from about 512 species have been recorded
from Canada to Argentina, being the Neotropical countries the ones
with the highest number of species. Among these Colombia with 163
species, is second after Brazil, with 242. In order to update the
information about Phlebotominae distribution in the Colombian
Orinoquian and Amazon region, records of species by locality were
obtained from collections made since the early sixties, and classified
according to Galati, 2003. Data from103 locations were collected with
an overall record of 92 species, 57 of them with exclusive presence on
the Amazon and/or Orinoquian region. The number of species per
genus was Bichromomyia (3), Brumptomyia (2), Evandromyia (8),
Lutzomyia (8), Micropygomyia (8), Migonemyia (1), Nyssomyia (5),
Pintomyia (6), Pressatia (4), Psathyromyia (15), Psychodopygus (15),
Sciopemyia (4), Trichophoromyia (9), Trichophygomyia (5),
Viannamyia (2). The species with the widest distribution was N.
antunesi. Among the species with epidemiological importance in
leishmaniasis we have N. umbratilis, N. yuilli, N. antunesi, B.
flaviscutellata, B. olmeca bicolor, B. reducta, P. davisi and P. hirsutus;
the first three with high abundance, anthropophilic behavior and
presence within the households. In conclusion, Phlebotominae
biodiversity in the Orinoquian and Amazon regions of Colombia is
high. It adds 57 species to the total list of species recorded in Colombia
and shares 102 taxa with records from other regions in the country,
some of them located in the genus Nyssomyia, Bichromomyia
Psychodopygus with vectorial background, mainly in cutaneous
leishmaniasis where this clinical form represents 23% of cases in the
country.
139
1
Centro Internacional de Zoonosis (CIZ) Universidad Central del Ecuador, Quito,
Ecuador, 2The Kuvin Center for the Study of Infectious and Tropical Diseases,
Department of Microbiology and Molecular Genetics, The Institute of Medical
Research Israel-Canada, Faculty of Medicine, The Hebrew University of Jerusalem,
Jerusalem, Israel. 3Unit of Molecular Parasitology &Tropical Medicine, Centro de
Biomedicina, School of Medicine, Universidad Central del Ecuador, Quito-Ecuador.
4
Prometeo, Secretaría Nacional de Educacion Superior, Ciencia, Tecnologia e
Innovacion (SENESCYT), Ecuador. 5Laboratorio Genética de Poblaciones, Sección
Invertebrados, Dept. de Estudios Ambientales, Univ. Simón Bolívar, Venezuela;
alonw@ekmd.huji.ac.il
Keywords: Cutaneous Leishmaniasis, Lutzomyia trapidoi,
Leishmania naiffi
In Ecuador cutaneous leishmaniasis (CL) is endemic and common in
most regions of the country including the western slopes of the Andes,
Andean valleys and the Amazonian lowlands. Several Leishmania
spp. cause CL in Ecuador including L.(Viannia) guyanensis, L. (V.)
panamensis, L. (V.) brazilensis, L. mexicana and L. major-like
parasites. We are conducting a project designed to study and
characterize the epidemiology of CL in several communities in the
district of Pichincha on the Western slopes of the Ecuadorian Andes,
where increasing numbers of CL cases are being reported. Although
the main aim of the project is to optimize diagnosis and treatment of
human CL cases by local clinics, we are also performing
entomological studies with the aim of devising disease prevention
strategies through sand fly control and curtailment of human-sand fly
contact. Here we report preliminary findings on sand fly species
altitudinal distribution, Leishmania infections and blood source
identification of sand flies in and near rural communities. The most
common sand fly was Lutzomyia trapidoi which was found in all
altitudes (200-1700 m above sea level) and habitats. The blood hosts
of 16 females were successfully identified using cytochrome b PCR
followed by DNA sequencing: 13 human, 2 opossum (probably
Didelphis marsupialis), 1 Porcupine (Coendou quichua). In parallel,
Leishmania infections were detected using ITS1 PCR followed by
DNA sequencing. Leishmania DNA was positively identified in 9
females (6 Lu. trapidoi, 2 Lu. panamensis, 1 Lu. hartmanni) out of 77
examined which is a very high infection rate. All DNA sequences
were of Leishmania (Viannia) spp. including L.(V.) braziliensis,
L.(V.) panamensis, L.(V.) guyanesis and L.(V.) naiffi. This is the first
record of L.(V.) naiffi from the western part of Ecuador. Studies are
ongoing and more data will accrue in the near future.
Acknowledgements: This work was funded by Fundação de
Assistência Médica Internacional (AMI), Portugal and Prometeo
Project of the Secretaria Nacional de Educacion Superior,
Ciencia,Tecnologia e Innovacion (SENESCYT), Ecuador.
N° 26 (1) 2015
ABUNDANCE OF PHLEBOTOMINAE IN ENDEMIC
AREA OF CUTANEOUS LEISHMANIASIS, TUCUMÁN,
ARGENTINA
PHLEBOTOMINE SANDFLIES (DIPTERA:
PSYCHODIDAE) OF MOROCCO: RESULTS OF
E N TO M O L O G I C A L S U RV E Y S A L O N G T H E
O U A R Z A Z AT- M ' H A M I D , Z A G O R A CHEFCHAOUEN-NADOR AND ZAGORA-TATAMARRAKESH ROADS
Direni-Mancini JM, Fuenzalida AD, Saracho-Bottero MN, Lizarralde de
Grosso MS, Salomón OD, Quintana MG
Instituto Superior de Entomología – UNT, REDILA
josemdireni@gmail.com
Keywords: Migonemyia migonei, Nyssomyia neivai, Evandromyia
cortelezzii, meteorological variables
140
Ouanaimi Fouad1; Boussaa Samia1,2; Kahime Kholoud1; Echchakery
Mohamed1; Boumezzough Ali1
Equipe d'Ecologie Animale-Environnement, Faculté des Sciences Semlalia, Université
Cadi Ayyad, Marrakech, Maroc; 2ISPITS-Institut Supérieur des Professions
fouad.ouanaimi@yahoo.fr1
In Argentina, 30 species of sandflies were recorded, which belong to
13 genera. Cutaneous leishmaniasis (CL) cases have increased
gradually in the province of Tucumán as well as in the remaining
endemic region. The last two outbreaks took place in 2003 and 2004.
Nyssomyia neivai is the main species incriminated in the transmission
of Leishmania braziliensis, parasite responsible of CL in the
northwest region of Argentina. The aims of this research were to
compare the abundance of Phlebotominae species in different sites
and seasons in addition to determine the meteorological variables that
have influenced in the abundance of the species in the south of the
province (27º35´05.89´´S - 65º37´11.70´´W).The study area belongs
to Yungas rainforest region, which has been currently modified for
different culture types. Five sample sites were selected based on
entomological and epidemiological antecedents. The sandflies were
captured with CDC light traps monthly for two consecutive days from
July 2008 to July 2009. The followings meteorological variables were
registered on the day of capture: Temperature (TºC), Relative
humidity (Rh%), Precipitation (Pp), Velocity of wind (Vw) and
Maximum velocity of wind (MVw). A total of 6793 individuals were
captured belonging to the species Migonemyia migonei (50.07%), Ny.
neivai (49.20%) and Evandromyia cortelezzii (0.74%). The
abundances of the three species were significantly different. Three out
five sites were the most abundant. Summer was the most abundant
season for the three species, followed by fall, spring and winter. The
main variables which affected the abundance of Mg. migonei were the
TºC and MVw (R2= 0.87); for Ny. neivai were the Rh% and MVw
(R2= 0.75) and for Ev. cortelezzii was the TºC (R2= 0.45). Mg.
migonei was more abundante in summer and spring (highest seasons
of activity for sandflies) whereas Ny. neivai, primary vector, was more
abundant in fall which is the highest transmission season. The three
most abundant sites are located at the outbreak area of 2003, where CL
cases were concentrated. The information provided in this study
contributed to the knowledge of the distribution of abundance in time
and space of CL vectors in northwestern Argentina.
Keywords: Sand flies, transects, entomological surveys, Morocco
Phlebotomine sand flies (Diptera: Psychodidae) are the vectors for
leishmaniasis as well as for arboviruses and bartonellosis.
Leishmaniasis are endemic in Morocco with two described forms
(visceral and cutaneous). Knowledge of vectors populations'
distribution is important in predicting the spatial variations in the risk
of disease. Previous studies in Morocco showed that the distribution
of sand flies was due, in great part, to the bioclimate. Current findings
in Morocco showed that altitude (through the gradient on
temperature, pressure and precipitation) and aspect (through climate
and vegetation) have an influence upon the spatial distribution and
density of the sand fly fauna. In the present work, we discuss the
possible effect of many ecological factors on the diversity and
distribution of sand flies in southeastern Morocco along three
transects (Ouarzazat-Mhamid; Zagora-Chefchaouen-Nador and
Zagora-Tata-Marrakesh). We give a particular attention to ZCL foci
(Ouarzazat, Zagora, Tata and Errachidia) with the aim to update their
entomological data.
Among the 2056 sandflies collected, in June
2010, on the Ouarzazat-M'hamid road, Phlebotomus longicuspis was
the most common species (25.5%), followed by P. papatasi (23.8%)
Sergentomyia fallax (12.6%), P. sergenti (16.6%), S. minuta (11%), P.
bergeroti (5.4%), P. dreyfussi (1.8%), P. christophersi (1.7%) and P.
ariasi, P. perniciosus, P. alexandri, P. chabaudi and P. chadlii with
less than 1% each. On the Zagora-Tata-Marrakesh road, P. papatasi
was the most prevalent species (24.2%) of the 2425sandflies that were
collected in June 2011, followed by P. longicuspis (17.3%), S. minuta
(16.4%), P. sergenti (11.9%), S. fallax (5.2%), P. dreyfussi (3.1%), P.
christophersi (2.8%), P. bergeroti (2.6%) and P. alexandri (1.2%). We
collected as well P. africana, P. chabaudi, P. ariasi and P. chadlii with
less than 1% each. In June 2012, 2828 sandflies were collected on the
Zagora-Chefchaouen-Nador road. P. papatasi was the most prevalent
species (33.5%), followed by P. sergenti (25.2%), P. longicuspis
(15.1%), P. perniciosus (6%), S. fallax (5.7%), S. minuta (4.7%), P.
alexandri (2.5%), P. chadlii (1.7%), P. bergeroti (1.5%) and P.
dreyfussi (1.5%). S. Africana, P. christophersi, P. chabaudi and P.
ariasi with less than 1% each. The distribution of potential vectors
along the three transects, according to altitude and bioclimate, was
explored.
N° 26 (1) 2015
141
S PAT I A L R E L AT I O N S H I P B E T W E E N
ENVIRONMENTAL FACTORS AND SAND flY
(DIPTERA: PSYCHODIDAE) DISTRIBUTION
ASSOCIATED WITH LEISHMANIASIS RISK IN
CENTRE & SOUTHERN MOROCCO
IDENTIFICATION AND CHARACTERIZATION OF
LARVAL LODGING OF PHLEBOTOMINE SAND
FLIES (DIPTERA : PSYCHODIDAE) IN MARRAKECH,
MOROCCO
Boussaa Samia1, 2; Ouanaimi Fouad1; Boumezzough Ali1
Kahime K, Boussaa S, Ouanaimi F & Boumezzough A
Faculté des Sciences Semlalia, Université Cadi Ayyad, Marrakech, Morocco
1
Equipe d'Ecologie Animale-Environnement, Faculté des Sciences Semlalia,
Université Cadi Ayyad, Marrakech, Maroc; 2ISPITS-Institut Supérieur des Professions
Keywords: SIG, Leishmaniasis, sand fly, risk, Morocco
Leishmaniasis, a highly neglected disease, currently presents a
significant health problem throughout Africa. In Moroccan context,
both cutaneous (CL) and visceral leishmaniasis (VL) have been
reported. CL is caused by three clinically important Leishmania
species (L. major, L. tropica, and L. infantum), a flagellate protozoa of
the family Trypanosomitidae, while, VL is caused by L. infantum.
Recently, the Mucocutaneous form was also reported but remains rare.
Leishmania infection is transmitted to human host as a result of a bite
by an infected female sand fly (Diptera: Psychodidae, Phlebotominae)
of the genus Phlebotomus. Over the past decade, the epidemiological
situation of CL has significantly increased with its geographic
expansion to previously free areas and the emergence of overlapping
foci of CL and VL in several provinces of Morocco. In order to
determine Leishmaniasis risk membership in central- south Morocco,
Geographic Information System (GIS) was used to investigate the
current spatial distributions of sand fly-borne pathogen risk. The
study was conducted in 69 sites where altitude varies from 14 to 2123
m. These sites belong to two administrative areas, the region of
Marrakech-Tensift- Al Haouz and the region of Souss-Massa-Draa. In
both regions, climate and vegetation are typically Mediterranean, with
hot, dry summers and maximum rainfall recorded in autumn and
spring. By sticky traps, we collected a total of 15 313 specimens,
belonging to 15 species of which10 species of the genus Phlebotomus
(57,38%). Proven and suspected Leishmania vectors were well
presented, in our study area, such Phlebotomus (Larroussius)
perniciosus Newstead (14,35%), Phlebotomus (Phlebotomus)
papatasi Scopoli (14,06%), Phlebotomus (Paraphlebotomus) sergenti
Parrot (12,85%), Phlebotomus (L.) longicuspis Nitzulescu (10,74%)
and Phlebotomus (L.) ariasi Tonnoir (2,68%). Sand fly repartition
linking to contextual environmental factors including geology, soil,
vegetation and climate were discussed for each species.
Keywords: sand flies, un-rotated genitalia, larval lodging,
Marrakech
Phlebotomine sand flies are the only vectors known of leishmaniasis.
These diseases are a serious increasing public health problem in
Morocco. In the aim to identify and characterize sand fly breeding
habitats in Marrakech city, our entomological investigations were
carried out during one year study in two sites (Akioud and Semlalia).
Using sticky paper, specimens were collected continuously between
October 2002 and September 2003 in Akioud, and from July 2002 to
July 2003 in Semlalia. Newly eclosed male sand flies were detected
based on their un-rotated genitalia. A total of 3571 specimens was
trapped during this entomological survey. Phlebotomus papatasi was
the most abundant species (53,2%), followed by Sergentomyia
minuta (20,1%), S. fallax (11,5%), P. sergenti (10,7%) and P.
longicuspis (4,5%). The sex ratio was in favour of males for all
species with the exception of P. sergenti (61.4% of females) in Akioud
and P. longicuspis (50% of females) in Semlalia. In Marrakech,
breeding habitats were identified for three, of five collected species,
as well as their emergence phases. Our results show that Akioud can
be breeding habitats for two Sergentomyia species (S. minuta and S.
fallax) while, Semlalia was identified as breeding habitats for P.
sergenti.Soil sample was collected from both sites where newly
eclosed male sand flies were detected. These samples were used to
determine organic matter values, humidity and soil texture of each
larval lodging. Characterization of these soil samples shows some
ecological requirements of immature stages development for these
sand fly species.
N° 26 (1) 2015
LEISHMANIASIS IN MOROCCO: STILL A TOPICAL
QUESTION
SEASONAL FLUCTUATION OF PHLEBOTOMINAE
(DIPTERA: PSYCHODIDAE) IN THREE
DEPARTMENTS OF AN ENDEMIC AREA OF
CUTANEOUS LEISHMANIASIS IN JUJUY PROVINCE.
A PRELIMINARY REPORT
Boumezzough A, Kahime K, Boussaa S & Ouanaimi F
142
Faculté des Sciences Semlalia, Université Cadi Ayyad, Marrakech, Morocco
aboumezzough@gmail.com
Keywords: Leishmaniasis, Epidemiology, Vector, Sandfly,
Morocco
Remondegui CV1,2, Cabrera CHA2, Quintana MG2,3
1
Cátedra de Biología Celular. Facultad de Ciencias Agrarias – UNJu, 2 REDILA, 3
Instituto Nacional de Medicina Tropical – MSN, Instituto Superior de Entomología
UNT-CONICET
crislr@gmail.com
Leishmaniasis is a parasitic disease a wide range of clinical symptoms
which currently threaten 350 million persons in 88 countries. It caused
by three Leishmania species, which are endemic, widespread and
represent a public health problem in most countries in the
Mediterranean basin. In Morocco, Leishmaniasis are endemic
diseases constituting a major public health threat. Cutaneous
leishmaniasis (CL) is caused by three clinically important Leishmania
species (L. major, L. tropica, and sporadic CL cases due to L.
infantum, flagellate protozoa of the family Trypanosomitidae).
Visceral Leishmaniasis is also caused by L. infantum. This study
presents a summarized analysis of its epidemiology in Moroccan
context. And aims to determine the current leishmaniasis
epidemiological situation in Morocco and the distribution of its
different forms throughout the country. During the past 20 years, this
disease has emerged as a major public health threat in Morocco. So,
we gave a particular attention to vectorial status of Moroccan sandflies
(Diptera: Psychodidae, Phlebotominae) in view of its major role in
diseases spreading. It seems clear that the risk of spread of the disease
is rising in Morocco. Ecological characteristic of proven and potential
vectors should be regarded as good marker to anticipate leishmaniasis
distribution in Morocco.
Keywords: Phlebotominae, Jujuy, Argentina
The endemic area of cutaneous leishmaniasis (CL) in the province of
Jujuy includes eight departments. The number of recorded cases of
leishmaniasis over the period 1990-2008 allowed ranking the risk
areas to contract the disease, placing the area of "Ramal" as the
greatest area of effective vector-parasite contact risk, concentrating
almost 50% of cases in the Ledesma Department. In 2008 we started
systemized studies to determine the presence of sandflies and were
recorded specimens of Nyssomyia neivai (Pinto), Migonemyia
migonei (Franca) and complex cortelezzii (Evandromyia cortezzii,
Ev. sallesi). The aim of this work was to study the seasonal fluctuation
of Phlebotominae species in three departments of the endemic area of
the province of Jujuy, specially in sites with epidemiological
antecedents. We conducted monthly sampling by 24 months from
September 2011 in selected sites of Ledesma, Santa Barbara and San
Pedro departments. The captures were made with CDC light traps by
two consecutive nights. The specimens captured were clarified and
preserved in eugenol until its identification to specific level. A total of
12013 sandflies were captured, a 51.5% corresponded to the Santa
Barbara department, the 30.5% to Ledesma department and 18% to
San Pedro department. In the three departments were recorded
abundance peaks in autumn and spring. Preliminary results obtained
in the present work show a increased abundance of sandflies in
autumn and spring with a decrease in the abundance in summer and
winter. The information resulting from this work on the diversity,
abundance and distribution of Phlebotominae in different seasons and
types of environments could be useful to detect effective measures
aimed at reducing the population of these insects, avoiding the risk of
outbreaks.
N° 26 (1) 2015
143
SPATIAL ANALYSIS OF THE OCCURRENCE OF
LEISHMANIASIS IN PARANÁ, BRAZIL, TO IDENTIFY
AREAS OF RISK
DENSITY DEPENDENCE IN HOST-VECTOR
RELATIONSHIPS AND CONSEQUENCES FOR
TRANSMISSION
Alceu Bisetto Júnior, Maurício Bisetto, Sônia Maria Dotto Ampessam
Erin Dilger, Orin Courtenay and Graham Medley
Secretaria de Estado da Saúde do Paraná
University of Warwick, UK.
abisetto@terra.com.br
erin.dilger@warwick.ac.uk
Keywords: Geoprocessing, American Cutaneous Leishmaniasis,
Epidemiology
Keywords: host choice, density dependence, Lutzomyia
longipalpis
This study aimed to compare the epidemiology and geographical
distribution of American Cutaneous Leishmaniasis (ACL) in the state
of Paraná in the period between 2008-2012 to spatial data altitude and
climate of the state using geoprocessing tools to identify risk factors
and areas for the occurrence of this disease. The occurrence of ACL is
directly related to the introduction of susceptible man to places where
the natural cycle of the disease is active, with the presence of the insect
vector and animal reservoirs. For the survey of the historical series and
epidemiological analysis based on state data from the Information
System for Notifiable Diseases from the Ministry of Health of Brazil,
with the record of notifications and investigations of ACL was used,
covering the period 2008 - 2012. The calculation of epidemiological
indicators was performed with the Tabwin/DATASUS program. To the
geographical location of clinical cases, according to place of residence
and infection, and the co-relationships of the geographical distribution
of cases with the environmental characteristics of favorable climate
and altitude of the occurrence Phlebotomines analysis of spatial
disease distribution was performed, the map hypsometric the state of
Paraná and according to Köppen climate classification, processed in a
Geographic Information System - GIS, using the mapping function of
Tabwin and georeferencing tools available. The results confirmed that
regions with altitude level less than 600 meters from the sea and Cfa
climate, according to Köppen , are at higher risk of contracting.
Sandflies readily feed on a wide range of hosts, and the dynamics of
preference between available host types are likely to be pivotal in
determining the transmission of multi-host diseases, such as zoonotic
visceral leishmaniasis. Sandfly host preferences dynamics are,
however, poorly understood, with simple relationships
conventionally assumed between host density, vector density and
transmission. Using integrated modelling and fieldwork approaches,
the preference of sandflies for key host types (dogs, humans and
chickens) and associated force of infection was investigated over a
range of vector and host densities. Following trap optimization and
the minimisation of trapping bias, preferences for key host types over
a range of vector densities were observed over a period of seasonal
variation in Brazil. Preference fluctuations in response to changes in
host density were also observed following experimental
manipulation of chicken densities within sheds. These investigations
indicate non-linearity in the relationship between sandfly preference
upon different host types with vector and host density. Pheromone
mediated aggregation behaviour in conjunction with host
attractiveness may explain additional attractive effects at higher
vector and host densities. Such aggregation in association with
outdoor resting hosts may have far-reaching implications for
understanding and manipulating transmission, and potentially
indicate host density manipulation as a possible control measure.
N° 26 (1) 2015
GEOGRAPHICAL DISTRIBUTION OF NYSSOMYIA
NEIVAI (PINTO) AND NYSSOMYIA INTERMEDIA
(LUTZ & NEIVA) IN THE STATE OF SÃO PAULO,
BRAZIL
FIRST MEETING OF LUTZOMYIA LONGIPALPIS IN
RURAL AREAS RELATED TO OUTBREAK OF
CANINE VISCERAL LEISHMANIASIS IN THE
NORTHWESTERN REGION OF SÃO PAULO, BRAZIL
Claudio Casanovaa, Paloma Helena Fernandes Shimabukurob, Bruna
Almeida Bressianic, Thaís Fernanda Lazaric, Eunice Aparecida Bianchi
Galati d
Eduardo Bergo, Lucimar Nascimento, Vera Camargo-Neves
a
Superintendência de Controle de Endemias, SUCEN, Mogi Guaçu-SP; b Fundação
Oswaldo Cruz, Centro de Pesquisas René Rachou, FIOCRUZ, Belo Horizonte-MG, c
FUNDAP/SUCEN, Mogi Guaçu-SP, d Faculdade de Saúde Pública, Universidade de
São Paulo, USP, São Paulo-SP.
144
Superintendence Of Control of Endemic Diseases
vlfcn@hotmail.com
Keywords: Lutzomyia longipalpis, epidemiology, entomology
egalati@usp.br
Keywords: Nyssomyia intermedia, Nyssomyia neivai, Distribution,
Cutaneous leishmaniasis vectors
Records of the geographical distribution of both arthropod-borne
diseases and the vectors involved in their transmission are essential for
epidemiological surveillance as well as the accomplishment and
formulation of control strategies. Nyssomyia neivai (Pinto) and
Nyssomyia intermedia (Lutz & Neiva), which comprise the
Nyssomyia intermedia s.l. complex, do not completely meet the
criteria to be incriminated as vectors of American cutaneous
leishmaniasis (ACL). However, their anthropophilic behavior and
predominance in areas where autochthonous cases of ACL have been
occurred allows to consider them as the main suspected vectors in
areas where these two species are found in the Southeastern region of
Brazil, particularly in São Paulo State. In the present study we provide
updated information on the geographical distribution of N. neivai and
N. intermedia in the state of São Paulo, where more than 6,000 human
cases of ACL occurred in the last 10 years. A database with records for
the presence of N. neivai and N. intermedia in the municipalities of
São Paulo state was compiled, using more than 10 different sources
between 1943 and 2013 (i.e. literature, examination of specimens
deposited in entomological collections, entomological surveillance
data). Our data clearly shows that there are three distinct distribution
patterns for these vectors: an allopatric distribution of (i) N.
intermedia in municipalities situated in the coastal region of the
Paraíba river valley and of (ii) N. neivai in the vast area of the Atlantic
Plateau region; and (iii) a sympatric distribution in municipalities
situated in the high areas of Ribeira and Paraíba river valleys, where
both species could be acting as vectors of ACL. Geographical
distribution databases are useful to map epidemiological data and to
build predictive models of disease.
Lutzomyia longipalpis (Lutz & Neiva) is the main vector of
Leishmania infantum chagasi (Cunha & Chagas), the etiological
agent of visceral leishmaniasis in the State of São Paulo (SPS). This
sandfly is well adapted to the peridomestic environment of the urban
areas of the municipalities of West Paulista Plateau, Western Region
of SPS. Since its first meeting in 1997, L. longipalpis has always been
associated with the VL transmission only in urban environments in
the state. This is the first reported finding of L. longipalpis in a rural
area and the record of canine visceral leishmaniasis(CVL).In June
and July 2013 were reported to the Superintendency of Control of
Endemic Diseases 11 dogs with suspected CVL in a rural area,
situated on the edge of the dam of the city of Pedregulho Estreito, at a
place called Água Azul Ranch. The city of Pedregulho has 8118.4 km
² and is geographically located in the central part of the SPS and
occupies 2.27% of the total area of the state. It is situated near the
basins of the Paraná and Paraguai rivers and geographical coordinates
are 20 ° 26'34 "South, 54 ° 38'47" West. According to Köppen's
climate is Tropical Altitude, with dry winters and mild summers. He
is presenting the rainy season is during the summer months
(November to March). The maximum average annual temperature is
28.5 °C and minimum of 9.4°C in the winter months (July-August).
The average annual rainfall is 1545 mm of rain. The entomological
captures were performed onsite Agua Azul Ranch, where these dogs
lived.In October and November 2013, considered one of the greatest
periods of vector chance meeting. Five light traps bait (AIL) model
modified CDC (Center for Disease Control) were exposed for three
consecutive weeks, and installed at five different points peridomicile
(next to the kennel, chicken house and the house owner, the porch and
around the tank water) As a result 43 individuals were captured:
53.5% were male and 5 species of sandflies were identified:
Nyssomyia whitmani (44.4%), Evandromyia lentivirus (22.2%),
Lutzomyia longipalpis (18,5%), Evandomyia cortellezzi (7.4%) and
Psychodopygus aragaoi (7.4%). Although most species caught were
preserved in this environment both Lutzomyia longipalpis and
Nyssomyia whitmani (the main vector of cutaneous leishmaniasis in
ESP) were captured in greater abundance in peridometic
environments and close to shelters for pets (kennel and chicken
coop). Contrary to what occurs in urban areas of the municipal.
N° 26 (1) 2015
STUDY OF SAND FLY COMMUNITY IN TWO RURAL
ENDEMIC AREAS FOR AMERICAN CUTANEOUS
LEISHMANIASIS IN THE MUNICIPALITIES OF MOGI
GUAÇU AND MOGI MIRIM, SÃO PAULO STATE,
BRAZIL
DNA DETECTION OF LEISHMANIA SP. IN FEMALE
SAND FLIES (DIPTERA: PSYCHODIDAE) IN BELO
HORIZONTE CITY AND SUMIDOURO STATE PARK,
MINAS GERAIS – BRAZIL.
1
2
Fernanda Elisa Colla-Jacques , Claudio Casanova , Fredy Galvis
Ovallos3
1
Departamento de Biologia Animal, Instituto de Biologia, Universidade Estadual de
Campinas, Campinas-SP; 2 Superintendência de Controle de Endemias, Mogi GuaçuSP, 3Programa de Pós Graduação em Saúde Pública, Faculdade de Sáude Pública,
Universidade de São Paulo, São Paulo-SP.
145
Saraiva, L.; Rugani, J.M.N.; Lima, A.C.V.M.R.; Reis, A.S.; Pereira,
A.A.S.; Gontijo, C.M.F.; Andrade Filho, J.D.
Grupo de Estudos em Leishmanioses/Centro de Pesquisa René Rachou/FIOCRUZ,
Belo Horizonte, MG.
jeronimomnr@hotmail.com
Keywords: Belo Horizonte city, Leishmania infantum, Lutzomyia
longipalpis
Keywords: Ny. whitmani, vectors, Leishmaniasis
Historically, a significant number of American cutaneous
leishmaniasis (ACL) cases registered in São Paulo State originated
from the Mogi Guaçu River Valley, and generally, the sandfly vectors
are abundantly present in this area. Based on these informations, this
work aimed to study the sandfly community in two rural endemic
areas for ACL in the municipalities of Mogi Guaçu and Mogi Mirim,
in the North-east region of São Paulo State, by analysing the spatial
and time distribution of the species in four different environments.
During the study period, from November/2010 to October/2011, CDC
electric light traps were installed every 10 days in two study areas area
from 18:00 to 06:00 hours. In each one of the study areas, four traps
were set up: one in the wooded area, one at the woods' edge, one in the
peridomicile associated with animals (chicken shed) and one in the
peridomicile associated to humans (porch). A total of 45,527 sandflies
were collected, comprised of 20 species. Of this total, 60% were
Nyssomyia whitmani, followed by Migonemyia migonei (19%) and
Expapillata firmatoi (7%). Among the studied environments, chicken
sheds were the most productive ones, where 85% of all the specimens
were collected. To understand the ecology of these species, several
indices were used to describe the sandfly communities. The chosen
indices were species richness, diversity, similarity and standardized
species abundance. The identification and dominance of Ny.
whitmani, Mg. migonei and Ex. firmatoi, suspected vector of ACL
agents, represents an increased risk for new cases.
Visceral Leishmaniasis (VL) have high incidence and mortality in
Belo Horizonte city and its metropolitan region. The present study
evaluated the detection of Leishmania sp DNA in female sand flies at
two area of cerrado biome know as Brazilian Savannah, one place is
an urban area in Venda Nova Sanitary District, in Belo Horizonte, and
a wild area of Sumidouro State Park, an environmental protected area
from 50 Km of Belo Horizonte. The samples were collected monthly,
during three consecutive days, from August 2011 to August 2013.
Eighteen traps were used in Venda Nova Sanitary District, and twelve
in Sumidouro State Park. Sand flies were identified following the
classification proposed by Galati 2003. Female sand flies were
subjected to DNA extraction procedures and tested for Leishmania
DNA using the ITS-1 molecular target. Positive samples were
sequenced for Leishmania species identification. Twelve females of
Lu. longipalpis species were identified with positive detection for
ITS-1 in Venda Nova Sanitary District, one being identified as
Leishmania braziliensis, four as Leishmania infantum, and the others
as trypanosomatids. Four females were positive at Sumidouro State
Park, one of cortelezzi Complex with positive detection for Le.
braziliensis, one of Psathyromyia lutziana species, one of Sciopemyia
sordelli species positive for Chritidia sp. and one specimen of
Evandromyia sallesi positive for trypanosomatids. The results shows
that the detection of sand flies infected with Leishmania sp. was
higher in urban ambient, with predominance of Le. infantum, which
indicates an increased risk of transmission in this area.
N° 26 (1) 2015
ID 176-P Control
ASSOCIATION OF OCCURRENCE AND ABUNDANCE
O F L U T Z O M I YA L O N G I PA L P I S W I T H
ENVIRONMENTAL VARIABLES AT DIFFERENT
SPATIAL SCALES: MICRO AND MACRO-SCALE
A PUBLIC HEALTH APPROACH TO LEISHMANIASIS
CONTROL
Santini MS1,2; Utgés ME2,3; Salomón OD2,3,4
1
Centro Nacional de Diagnóstico e Investigaciones en Endemoepidemias-ANLIS-MSN,
Argentina; 2 REDILA,Argentina; 3 Instituto Nacional de Medicina Tropical-MSN,
Argentina; 4 Consejo Nacional de Investigaciones Científicas y Técnicas.
Keywords: Lu. longipalpis, occurence and abundance, visceral
leishmaniasis
Lutzomyia longiplapis is the species responsible of transmitting Le.
infantum (Syn. chagasi) in America. The objective of this study was to
evaluate the environmental factors that determine the occurrence and
abundance of Leishmania spp. vectors in urban settlements. The study
was conductedon during February 2013 in Santo Tomé City,
Corrientes, Argentina. Fifty-three households were selected within
each 600 m2 square using the “worst scenario” criterion. Sandflies
were captured with mini-CDC light traps, active for 3 consecutive
rainless nights. Environmental variables were gathered at two spatial
scales: micro and macro-scale. At each household (micro-scale), a set
of 6 variables were gathered at the same time of the entomological
sampling (Trees near the trap, Fruit trees, Plant pots, Dogs, Hens and
Unused materials). Macro-scale variables where obtained from the
GPS (Altitude), or from a SPOT-5 image with 10 m of spatial
resolution (Distance to stream, Distance to city border, Proportion of
Urban cover, High and Low density vegetation cover, and Bare Soil
cover). To evaluate the effects on accumulated abundance of Lu.
longipalpis, we constructed a set of 13 models to account for micro-,
macro- and mixed-scale effects. Models were either GLM's with
negative binomial distribution, or Hurdle models. The final model was
selected by means of the AICc criterion. Parameter estimates and BCa
intervals of the final model where calculated by bootstrap based on
1000 replications. We captured a total of 853 sandflies belonging to six
species: Lu. longipalpis, Migonemyia migonei, Nyssomyia withmani,
Bruptomyia spp., Ny. neivai and Evandromyia cortelezzii-sallesi. The
98.35% of the sandflies captured were Lu. longipalpis. In relation to
environmental variables and according to the final hurdle model, only
Distance to the border of the city and High to medium density
vegetation cover ended to be positively associated with the occurrence
of sandflies in the city (macro-scale variables). Trees around the trap,
Distance to the stream and its cuadratic, were the variables positively
associated with the abundance of sandflies (micro and macro-scale
variables). These results suggest that presence and abundance of Lu.
longipalpis could be explained by different factors depending on the
spatial scale considered. In these work, presence and abundance of
sandflies were defined by different spatial scales.
146
Waleed Al Salem1,2*, Badriah Al Otaibi2,3, Louise Kelly-Hope1,4, Karina
Mondragon-Shem 1 , Salah Balgonaeem 2 , Maha Abdeladhim 5 ,
Mohammed Al Zahrani2, Amir Hassan3, Jesus Valenzuela5, Alvaro
Acosta-Serrano1,6
1
Parasitology Department, LSTM, England, 2Saudi Ministry of Health, Riyadh, Saudi
Arabia, 3International Public Health, LSTM, R England, 4Neglected Tropical Disease
Centre, LSTM, England, 5 Vector Molecular Biology Section, National Institute of
Allergy and Infectious Diseases, National Institutes of Health, Maryland, USA, 6Vector
Biology Department, LSTM, England
waleed-alsalem@hotmail.com
Keywords: Old world cutaneous leishmaniasis, Saudi Arabia, Al
Ahsa
Old World cutaneous leishmaniasis (CL) is one of the most prevalent
vector borne diseases in the East Mediterranean Region (EMRO). A
combination of factors is responsible for the sustained spread of CL
throughout this region, including urbanization, irrigation,
governmental sector integration, socio-economic factors, lack of
health education, civil war and human migration. Civil war in several
nations in the EMRO region, including in Afghanistan, Syria, Yemen,
Iraq, Libya and Lebanon, and mass human displacement triggered by
these conflicts are responsible for the current spread of CL to new
foci. Since the beginning of the civil war in Syria approximately 2.7
million Syrians have become refugees in Turkey, Lebanon, Jordan,
Iraq and Egypt. Moreover 4.1 million Syrians have been internally
displaced within Syria. Another consequence of conflict is the lack of
leishmaniasis control activities, which can further exacerbate CL
spread. Uncontrolled urbanization and labour migration has led to the
appearance of several new foci in Saudi Arabia, made worse by the
lack of a health impact assessment for the new settlements.
Settlements in Northern Israel have experienced CL outbreaks
because of their proximity to leishmaniasis endemic areas. Irrigation
may also create conditions favourable to the establishment of new CL
foci, by changing the surrounding environment of areas, such as the
Sidi Saad Dam in Tunis. Notably, foreign troops stationed in the
EMRO region tend to be highly affected by CL due to a lack of
immunity to the parasite, especially in CL endemic areas. In Iraq, an
estimated 2500 overseas soldiers have been infected by CL, of whom
1283 soldiers presented in 2003 following their tour of duty. Also the
International Security Assistance Force (ISAF) reported 200 cases of
CL at its bases in Mazar-e Sharif, Afghanistan. New control
measures, based on a systematic public health approach, are urgently
required to minimize the impacts of conflict and mass migration on
CL spread. This study was carried out in the Al Ahsa governorate,
located in the eastern region of Saudi Arabia. Al Ahsa is a large oasis
surrounded by three deserts, Al Dahna, Al Summan and Empty
Quarter, and has an estimated population of 1,165,422 people,
comprised of 80% Saudis and 20% non-Saudis. Regarding health
infrastructure, Al Ahsa has 18 hospitals and 68 primary health care
centres. There are also three leishmaniasis clinics offering diagnosis
and treatment: Al Yahya for the norther.
N° 26 (1) 2015
147
N° 26 (1) 2015
ISOPS VIII
Country
1
Argentina
2
Bolivia
3
Brazil
4
Canada
5
Colombia
6
Authors
Orals
52
Posters
1
6
326
24
2
10
86
42
8
6
Czech Republic
17
6
5
7
Ecuador
10
1
1
8
Ethiopia
6
4
2
9
France
13
2
10
Gabon
1
11
Germany
3
12
Guatemala
1
13
India
13
4
1
14
Iran
13
1
1
15
Israel
9
4
2
16
Italy
29
3
1
17
Japan
4
1
18
Kazakhstan
6
19
Madagascar
2
20
Mexico
6
1
21
Morocco
5
5
22
Nepal
3
23
Panama
8
24
Paraguay
8
25
Peru
26
Portugal
9
27
Saudi Arabia
8
2
1
28
Spain
25
3
6
29
Sudan
4
1
30
Switzerland
2
31
Tunisia
47
6
4
32
Turkey
23
2
3
1
24
1
1
1
1
1
1
1
1
6
3
148

Boletín de la SEA - Sociedad Entomológica Argentina | SEA

Transcription

Similar documents

Contents - Sand Sports Magazine

FLEXIRIDE™ - Equestrian Direct

cave painting - Sakura Of America

Leishmaniasis Factsheet

Is play sand a health hazard?

available for download.

Leishmaniasis in the WHO European Region

View Product Label

Geological models, heterogeneities and scale relations

Acrobat1.2MB