Issue 6 , December 2012 - CRIP

A Message from the Director
2012 was a busy year which was rich in emotion for the
Centre! First, we hosted two events during the spring: the
Café-CRIP in April with the valued collaboration of the
Centre québécois de valorisation des biotechnologies and
our annual Symposium in May. These activities are
important for the CRIP for several reasons. It encourages
networking between our members, our students and our
partners in the agri-food industry; it demonstrates our
expertise and ability to lead innovative projects that
generate solutions for the industry; and it exposes the
Centre to national and international scientific community.
Our calendar for 2013 is already completely booked. As
expected, everyone will be asked to participate and
collaborate at our future meetings.
The road towards CRIPA
During the summer, the preparation of our application to renew our Fonds de recherche du
Québec-Nature et technologies (FRQ_NT) Strategic Cluster funding has kept us busy. First and
foremost, we have a new name: CRIPA! And following this great team effort, I would like to
sincerely thank all the members for their involvement and constructive comments. This renewal
will breathe new life into our group by the addition of the poultry production to our scientific
program. This will add 10 new members to CRIPA and will raise our total regular and associated
membership to 40. The application was submitted on November 1st and will be presented in
January 2013 to the FRQ_NT visiting committee. We will then patiently wait for their decision.
With all these changes, I would like to sincerely thank some members that have left for other
positions or endeavours and those that have retired! Their scientific contribution, promotion of
the CRIP and participation in CRIP activities was remarkable and appreciated. Thank you!
I invite you to read this 6th edition of the Info-CRIP that highlights the activities of our research
group and its members. This is an edition that marks a new historical direction for our Centre.
Have a good read,
Josée Harel, director
Welcome to Our New Members
During the preparation of our application for renewal, we recruited several individuals to become
members of our strategic cluster. These new recruits will enrich and diversify the expertise in the
CRIPA. For some (*), their mandate will begin only with the renewal of CRIPA. We welcome
them and here is a brief description of the new members.
Maryse Boucher*, professor, CEGEP de Saint-Hyacinthe. She is the only college
researcher and is a microbiologist-immunologist. She teaches biology in several
programs including: nursing, animal health, dental hygiene, biotechnology, dietetic
and natural sciences. Since October 2012, her research has taken place at
Cintech agroalimentaire where she provides technical support in microbiology.
Email: mboucher@cegepsth.qc.ca.
Martine Boulianne*, full professor, Faculté de médecine vétérinaire, Université de
Montréal. Her research interest is poultry: with feathers, carcass and eggs. She
investigates the risk associated with Salmonella and Campylobacter
contamination. Alternatives to antibiotic use are also part of her research priorities.
Email: martine.boulianne@umontreal.ca.
Web page: www.medvet.umontreal.ca/sciences_cliniques/boulianne.htm.
Younès Chorfi*, assistant professor, Faculté de médecine vétérinaire, Université
de Montréal. His research interests focus on the effect of mycotoxins on the health
and performance of animals, the immune system and susceptibility to infectious
diseases. He is also interested in improving animal health through improved
nutrition and feed.
Email: younes.chorfi@umontreal.ca.
Caroline Duchaine, full professor, Université Laval. She has an interest in
environmental microbiology, specifically in the domains of aerobiology and
aerovirology. These domains study air-born particles of biologic and viral origin.
Email:caroline.duchaine@bcm.ulaval.ca.
Web Page: http://www.bioaerosols.ulaval.ca/equipe/
Sébastien P. Faucher, assistant professor, McGill University, Macdonald
Campus. His research interests are focused on host-pathogen interactions,
more specifically in the virulence factors and regulatory networks that are
involved in these interactions. His favorite bacterium is Legionella pneumophila.
Email: sebastien.faucher.2@mcgill.ca.
Web page: http://www.faucherlab.com/
Ismail Fliss*, full professor, Faculté des sciences de l’agriculture et de
l’alimentation, Université Laval. He is interested in the role of probiotics and
probiotic bacteria in the prevention and resistance to enteric infections. The use of
rapid detection methods of pathogenic microflora (DNA probes, PCR, RT-PCR,
NASBA) and in vitro simulation of the human digestive system are also part of his
research.
Email: ismail.fliss@fsaa.ulaval.ca.
Web page: www2.ulaval.ca/ulaval_ca/recherche/chercheur/fiche/241602.html
Info-CRIP No 6 – December 2012
2
Philippe Fravalo, associate professor, Faculté de médecine vétérinaire,
Université de Montréal. His main research interests are important pathogens
associated with food-borne infections and food-borne pathogen surveillance
in the poultry and porcine industries.
Email: philippe.fravalo@umontreal.ca.
Web page: www.medvet.umontreal.ca/professeurs/pfravalo.htm
Frédéric Guay*, full professor, Faculté des sciences de l’agriculture et de
l’alimentation, Université Laval. His research focuses on the management and
nutrition of pigs, vitamin and mineral nutrition, organic swine production and
mycotoxins.
Email: frederic.guay@fsaa.ulaval.ca.
Web page: www2.ulaval.ca/ulaval_ca/recherche/chercheur/fiche/364523.html.
Yvan L’Homme, researcher, Canadian Food Inspection Agency, Saint-Hyacinthe.
He works on the discovery, characterization and classification of new viruses, and
on epidemiology and control of enteric viruses in production animals and humans.
He is also interested in virology and food-borne viruses, vaccines, zoonotic
pathogens, viral indicators of contamination and microbial synergy.
Email: yvan.lhomme@inspection.gc.ca.
Nicholas Ogden, researcher, Public Health Agency of Canada. His interests are
on the ecology of Lyme disease and of other vector-borne diseases and zoonosis.
He is also interested in risk prediction associated with climate changes, and the
development of methods to improve surveillance and control of infectious
diseases. Email: nicholas.ogden@phac-aspc.gc.ca.
Xin Zhao*, full professor, McGill University. His research interests are focused
on microbial virulence factors that affect the host immune system, mechanisms
used by bacterial pathogens to initiate intestinal infections and the
development of vaccines, prebiotics and probiotics as alternatives to
antibiotics.
Email: xin.zhao@umcgill.ca.
Web page: http://www.mcgill.ca/animal/staff/faculty/zhao.
News from Our Members
In the last year, several members have had outstanding news and here is some of the news that
made it to us:
Denis Archambault, UQAM
Dr. Archambault and Andrea Gomez Corredor, a postdoctoral fellow at the Institut de
Recherches Cliniques de Montréal, published novel results in May in the prestigious Journal of
Virology. Their work characterized the pathway involved in the Rev
uptake pathways involved in the regulation of viral genes. This discovery
also identified new drug targets that could be developed to fight AIDS.
Steve Charette, Université Laval
Dr. Charette was awarded third place in the “La Preuve par l’image 2012”
contest hosted by Acfas with his picture entitled “La forêt des affamés”.
Info-CRIP No 6 – December 2012
3
Martin Chénier, McGill University
The work published in Microbial Ecology by Dr. Chénier and his team was the subject of an
opinion piece in Science on the persistence of antibiotic resistance in the environmental
microflora of pig farms.
news.sciencemag.org/sciencenow/2011/drug-resistance-loiters-on-antib.html.
Martine Denicourt, Université de Montréal
Dr. Denicourt was appointed Vice-president of the executive committee for the Association des
vétérinaires en industrie animale, AVIA.
J. Daniel Dubreuil, Université de Montréal
"The Whole Shebang: The Gastrointestinal Tract, Escherichia coli Enterotoxins and Secretion".
Dr. Dubreuil just published an extensive review on diarrhea caused by E. coli. This review
explores the interaction between pathogenic E. coli and the intestinal system which includes the
nervous system and the diverse factors leading to diarrhea. The review is available for those
interested in understanding the mechanisms involved in E. coli infection, the different toxins
produced and their effect on the gastro-intestinal system in clicking to this link:
http://www.horizonpress.com/cimb/v/v14/71.pdf.
Marcelo Gottschalk, Université de Montréal
Dr. Gottschalk is co-author of a clinical study on cirrhotic adults that have eaten uncooked pork
meat and showed S. suis infection that involved two new serotypes. In September 2011, the
research team published the results of this study entitled: "Sepsis and spontaneous bacterial
peritonitis in Thailand" in The Lancet, vol 378: 960.
The team of Dr. Gottschalk has secured funding from the Association of Universities and
Colleges of Canada for an International Development project between Canada, Latin America
and the Antilles. The funded project will collect data regarding infections caused by
Streptococcus suis, a pathogen of human and pig.
John M. Fairbrother and Éric Nadeau, Université de Montréal
Prevtec microbial is a spin-off business from the Escherichia coli Laboratory (Ecl) and is
lead by Drs. John M. Fairbrother and Éric Nadeau. Coliprotec, a vaccine developed
by the company, has been approved in Brazil. Prevtec has also signed an agreement
with the French multinational corporation, Virbac, to distribute the vaccine in France. « Prevtec
Microbia : Chez les tsars ou chez l'Oncle Sam ? ». lapresseaffaires.cyberpresse.ca/prevtecmicrobia-chez-les-tsars-ou-chez-loncle-sam-.php. Prevtec was also the subject of a new report
in La Presse as for an emerging innovative company: affaires.lapresse.ca/prevtec-microbia-visele-monde.php.
In July, Dr. Fairbrother received the ALUMNI Award 2012 from the University of Sydney in
Australia that recognised his exceptional contribution to the international scientific community.
Ann Letellier, Université de Montréal
Dr. Letellier has received the Vétoquinol award for research which recognises the contribution of
a member of the teaching staff to train graduated veterinary science students.
Mariela Segura, Université de Montréal
The article “Streptococcus suis Capsular Polysaccharide Inhibits Phagocytosis through
Destabilization of Lipid Microdomains and Prevents Lactosylceramide-Dependent Recognition”
by Mariela Segura and her research team which includes two other members of the CRIP, Dr.
Marcelo Gottschalk and Dr. Marie-Rose Van Calsteren, was published in the February edition of
Info-CRIP No 6 – December 2012
4
Infection and Immunity of the American Society for Microbiology. This article was selected for a
spotlight which highlights research of great scientific interest
especially for the novelty of the discovery and its scientific merit.
In September, Dr. Segura received a Women of Distinction award,
a prestigious award from the Women’s Y Foundation of Montréal.
The award recognizes her involvement in the domain of sciences
and technologies. Congratulation Mariela!
News from the Administrative Center
Good retirement Manon Coutellier!
Last June, the GREMIP honored Manon Coutellier, also secretary at the CRIP, for
her well-deserved retirement. Everyone thanks her for her good advice, her
meticulous and keen work, and her attention to detail that simplified the life of
GREMIP and CRIP members without forgetting numerous hours worked behind
the scenes. Her departure left a vacancy that has been occupied since June by
Isabelle Flibotte. Welcome Isabelle!
A birth at the CRIP!
At the beginning of June, Cécile Crost, coordinator at the CRIP, left for her maternity leave.
Since then, the birth of her son Paul has kept her busy! Cécile is back since December 10.
Anne-Marie Christen took over Cécile’s job during her leave. Anne-Marie’s mandate with the
CRIP will be done at the end of January 2013.
It’s official, the egg comes from the chicken!
On October 10th 2012, the Fédération des producteurs d’œufs de consommation du Québec
(FPOCQ) invited their partners to participate in a guided tour to learn about all the branches of
their industry. As a new partner, the CRIP was represented by the
author and, as a witness, she can now confirm that the egg comes
from the chicken. First, a bus brought around fifty participants to
Ferme avicole Marie Pierre which is located in Roxton Pond. This
family-owned farm produces eggs for human consumption. At the
farm, we were able to see all the equipment and facilities required
for egg production, including the heating system, automatic feeders,
egg collectors, manure management, biosecurity and the
preparation of the eggs to be shipped to the grading station.
About two days after the eggs are laid, the eggs are sent by
refrigerated truck to Nutri-Oeuf in Saint-Hyacinthe. At the plant, the
eggs are washed, candled, graded, cartoned, packaged and sent to
different supermarket chains and grocery stores. Several operations
are automated but a pair of eyes is always needed at the grading station to spot and remove the
cracked eggs.
The guided tour ended at Vitoeuf Inc, a family-owned business that transforms eggs into its
liquid and hard boiled forms.
We would like to thank the FPOCQ for this interesting visit and initiative!
Anne-Marie Christen, CRIP coordinator
Info-CRIP No 6 – December 2012
5
CRIP Activities in 2012
During the last year, several activities were organized by the CRIP to create networking
opportunities for students, members and our partners. Here is a summary of the events
highlighting the work of several CRIP members.
The 5th Symposium
Our 5th Annual Symposium was held on May 16 and 17 at the Faculté de médecine vétérinaire
(FMV) of the Université de Montréal. The event was possible due to the generous financial
support of PFIZER animal health, OLYMEL, ELANCO, MIA CELLAVIE Inc, BIOAMERICA,
QIAGEN and the FMV.
High quality conferences
A total of 125 participants were present for four
presentations that covered the field of immunology,
bacterial pathogenesis, virology and public health
which were presented by Drs. Volker Gerdts from
the University of Saskatchewan (VIDO), David
Francis of South Dakota University (USA), Hans
Nauwynck from Ghent University (Belgium) and
Wondwossen Gebreyes of Ohio State University
(USA), respectively.
Left to Right : David Francis, Hans Nauwynck,
Josée Harel, Volker Gerdts, Wondwossen
Gebreyes et Carl A. Gagnon
Dr. Martin Lessard from the Dairy and Swine
Research and Development Centre of Agriculture
and Agri-Food Canada in Sherbrooke presented
his latest results regarding the use of chitosan as
an antigen transporter in the intestinal tract.
Our Trainees
We were impressed by the quality of the oral presentations given by 14 students in the CRIP.
The 27 poster presentations were also of a high scientific calibre. A workshop on careers
associated with animal health was also given by Jean-François Marquis (Vertex Pharmaceutical)
and Sébastien Faucher (McGill University). This workshop was a success and appreciated by
the students.
Dr. Pascal Dubreuil, Vice-dean for Clinical Affairs of the FMV, presented the awards for the best
and second best oral and poster presentations.
Best oral presentations: 1st place: Damian Clarke, a
FMV student supervised by Dr. Mariela Segura, for his
presentation entitled “Role of CD4+ T cells in the
immune response against encapsulated Group B
Streptococcus”. 2nd place: Rémi Lavoie, a FMV student
supervised by Drs. Josée Harel and Christine Martin,
for his presentation entitled “Single‐cell measurement
of F165.1 and Pap phase variation in real‐time”.
Best Posters: 1st place: Bruno Haas a Université Laval
student supervised by Dr. Daniel Grenier for his poster
entitled “DNase activity of Streptococcus suis:
Left to right : Pascal Dubreuil, Damian
Clarke, Josée Harel, Rémi Lavoie, Chan Wu
and Bruno Haas
Info-CRIP No 6 – December 2012
6
Characterization and possible roles in virulence”. 2nd place: Chan Wu, a FMV student supervised
by Dr. Mario Jacques, for her poster entitled “Zinc as an agent for the prevention of biofilm
formation by pathogenic bacteria”.
Café-CRIP
Why and how to control biofilms in the agri-food industry?
Dr Mario Jacques from the CRIP teamed up with the Centre québécois de valorisation des
biotechnologies (CQVB) and its dynamic animal health Director, Mrs Dora Rodriguez, to host its
second Café-CRIP on April 24th. More than 80 participants from universities and the agri-food
sector interested in the impact of biofilms in the agri-food industry learned about the efficacy of
equipment disinfection and the treatment of infections with antibiotics.
Left to Right: MM. Michel Pouliot, Merle E. Olson,
Mario Jacques, Sylvain Fournaise, Phil Stewart,
Luc Lagacé and Dora Rodriguez from CQVB
The five speakers were Mr. Phil Stewart (Center
for Biofilm Engineering, Montana State
University, USA), Mr. Luc Lagacé (Centre ACER
inc., Québec), Mr. Merle E. Olson (Innovotech
inc., Alberta), Mr. Michel Pouliot (Agropur,
Québec) and Mr. Sylvain Fournaise (Olymel,
Québec). They presented on the challenges,
surveillance, innovation and protocols associated
with the control of biofilms in different agri-food
industries including the maple, milk and meat
transformation. The take home message was
that biofilms can be controlled but may not
always be eradicated. A summary was prepared
by the CQVB and is available on CRIP’s web site
(in French only).
To learn more about biofilms, you can obtain the French publication « BioTendance®: Les
biofilms: s'en préoccupe-t-on assez dans l'industrie agroalimentaire ? » produced by CQVB by
clicking the following hyperlink: Pourquoi et comment contrôler les biofilms dans l'industrie
agroalimentaire ? - Publication.
Lunch-Conferences of the GREMIP/CRIP
In the last year, the students, members and guests of the CRIP have had the opportunity to
acquire new knowledge during the nine lunch conferences hosted by the GREMIP/CRIP. These
interesting presentations offered an opportunity to explore new research field, and to chat in a
relaxed atmosphere with high level researchers.
A renowned visitor – Dr. Roy Curtiss III
In October, Dr. Roy Curtiss III presented a talk entitled
“Induction of Cellular Immunity and Delivery of DNA Vaccines
by Recombinant Salmonella” during the FMV research day in
association with the 30 year anniversary of the GREMIP.
Dr. Curtiss is the director of the Center for Infectious Diseases
and Vaccinology and Microbial Genetic Engineering at the Dr Roy Curtiss III, professor at
Biodesign Institute and he is also a leader in the field of genetic Arizona State University
and molecular pathogenicity of bacteria. The research goals of
the Center for Infectious Diseases and Vaccinology and Microbial Genetic Engineering are to
Info-CRIP No 6 – December 2012
7
develop vaccines to prevent enteric and respiratory infectious disease in humans and animals,
including poultry. Dr. Curtiss III’s work and discoveries have resulted in the publication of more
than 250 scientific articles and the creation of several patents.
Presentations given in the previous year are summarized in the following table:
Invited Speakers
Date
Titles
2011
Jean-Pierre Vaillancourt,
FMV, UdeM
November 17
Les Belles soirées de l’Université de Montréal
Recherche en biosécurité à la ferme /
Orientations futures
Steve Charette, U Laval
December 8
Modèle infectieux amibe et Streptococcus suis
2012
Luke Masson, IRB, CRNC
February 16
Ken Dewar, U McGill
April 12
Michel Desjardins, UdeM
May 10
Christian Baron, UdeM
June 5
Guy-Pierre Martineau,
École nationale vétérinaire
de Toulouse, France
June 8
Timothy Geary, U McGill
October 4
Roy Curtiss III, U of
Arkansas
October 17
Customized DNA Microarrays: Handy tools for
pathogen detection and source tracking
Intestinal microbiome variation in vervet monkeys
in a common environment with a controlled diet
La contribution de la protéomique à la
connaissance des fonctions du phagosome
Développement d’inhibiteurs des systèmes de
sécrétion comme médicaments antimicrobiens
Le GREMIP et les pathocénoses
Resistance to macrocyclic lactone preventatives
in the dog heartworm Dirofilaria immitis
Induction of Cellular Immunity and Delivery of
DNA Vaccines by Recombinant Salmonella
Workshops for Students
In addition to the annual symposium and the lunch-conferences,
several workshops are organised for students to help them
acquire new knowledge and diversify their skill sets.
Technical Workshops of the CRIP
Two technical workshops were held this year, one in statistics
and one in microscopy.
March 30 – Comprendre et appliquer les statistiques paramétriques en biologie
By Gabriel Perron, Ph. D., Department of System Biology, Harvard University
Twenty-two students and members of the CRIP benefited from Dr. Perron’s teaching during this
biostatics workshop. The t-test, the general linear models and linear regression models are no
longer mysterious for these students!
Info-CRIP No 6 – December 2012
8
September 27 2012 - Microscopie de fluorescence quantitative : aspects
pratiques
By Judith Lacoste, Ph. D., MIA CELLAVIE Inc.
This interesting and practical workshop gave an overview of the proper use of
microscope. Dr. Lacoste presented several practical tricks to ensure that good
quality images are acquired with a microscope. A PDF file is available on the
CRIP website for those that unfortunately missed this workshop. The file can be
obtained by clicking here:
www.crip.umontreal.ca/documents/documents/MicFluoAspectsPrat27sep2012_JLacoste.pdf
Bursaries from FPPQ, CRIP and FRQ_NT
Gaël Auray was awarded a $25,000 study grant from the
Fédération des producteurs de porcs du Québec!
Gaël is a post-doctoral fellow in the laboratory of Dr. Marcelo
Gottschalk at the FMV. He won a contest held by the FPPQ and he
was awarded a $25,000 study grant. Congratulation Gaël! And a
sincere thank for the financial support of the FPPQ!
Gaël Auray receiving his grant from Mr. Normand Martineau of the FFPQ
In 2011, this study grant was awarded to Fernando Alvarez, master candidate student in the
laboratory of Dr. Carl A. Gagnon, FMV, UdeM.
Need-based Bursaries
Master: Maryline Bouchard (UdeM), Guillaume Goyette-Desjardins (UdeM), Cristelle Karam,
(UQAM) and Jean-Philippe Auger (UdeM) were each awarded a $5,000 bursary.
Doctoral: Sébastien Crépin (IAF, INRS), Alexandre Thibodeau (UdeM), Virginie Lachapelle
(UdeM) and Bruno Haas (U Laval) were each awarded a $6,000 bursary.
Travel Grants
Eleven travel grants were awarded to the following students: Clément Muzika (UdeM),
Mohammed Chekabab, (UdeM), Jean-Philippe Côté (UdeM), Devin Holman (U McGill), Claude
Lachance (UdeM), Amélie Garénaux (IAF, INRS), Yannick Tremblay (UdeM), Skander Hathroubi
(UdeM), Christian Savard (UdeM), Fernando Alvarez (UdeM) and Yenney Hernandez Reyes
(UdeM). The next contest will be organised in February 2013.
FRQ_NT Postdoctoral Grant
In May 2012, Christian Savard, postdoctoral fellow in the laboratory of Dr. Carl A. Gagnon, was
awarded a $60,000 grant from FRQ_NT for a period of two years.
Congratulations to all our students!
Info-CRIP No 6 – December 2012
9
An international internship bursary from FRQ_NT
An internship in Japan
Why not?
“I greatly encourage student to learn about the international internship bursary. Several
interesting bursaries are offered every year by different agencies including the FRQ_NT. This is
a unique opportunity to have a great learning experience for students that they will remember all
their lives.” David Roy, student at the Faculté de médecine vétérinaire, Université de Montréal.
First and foremost, the choice of Japan for my
internship was based on a desire to acquire new
technical knowledge and to develop a new
collaboration between the laboratory of Dr.
Mariela Segura and the Japanese team of Dr.
Takamatsu. I have to say that for a first
international internship, Japan was the best
destination to get me out of my comfort zone
because Japan is the opposite of Quebec. I
jumped at the opportunity to do an international
internship because it offered a great
professional development opportunity and
allowed me to discover a country and a culture that is greatly different than my own.
When I arrived, the first thing I noticed was the cleanliness and the effort people put into
maintaining the country clean and, also their attention to detail. I then began to experience
homesickness and, the most difficult aspect, the language barrier. Few Japanese people speak
English. However, they were welcoming and their cheerfulness was contagious. These aspects
helped overcome the language barrier and the homesickness and this made the integration into
the research team and Japanese culture easier. During my stay, I lived in the international
student residence and this gave me the opportunity to meet other international students and
make new friends, including individuals from Thailand and India.
Training Received
Since my first internship at the GREMIP, I have worked
with several genetic tools developed by Dr. Takamatsu’s
group. My internship in Japan gave me the opportunity to
meet our collaborators. I sought to benefit from their
expertise in order to clarify several aspects of their
protocol and to ask for advice to help me navigate the
future steps of my research project. While I was there, I
tried to learn as much as possible about the genetic tools
the Japanese laboratory developed, which I use on a
regular basis in Quebec.
Given that I had a basic knowledge of the mutation vector developed by Dr. Takamatsu, I was
able to learn different applications for the vector series that are available. During my internship, I
developed plasmids to be used for interspecies gene-exchange for genes located in the
polysaccharide capsule encoding locus.
Info-CRIP No 6 – December 2012
10
Thus, this training reinforced my knowledge but also added value to the genetic tools I will be
using for different projects in Dr. Segura’s laboratory. When I had free time, I was also able to
work on Dr. Takamatsu’s personal project which involves a pathogen of bees.
The advantages of an international internship
Of course, an international internship is not limited to
working. Although Japanese are dedicated and hard
workers, I was able to use my free time to visit their
beautiful country. In Japan, 2-3 days is enough time to
visit several cities because their public transit is
extraordinary. However, certain places or islands are
only reachable by plane or boat and to visit these
several days off are required.
Despite the limited time to travel, I visited several cities
with different attractions and features. This allowed me to
appreciate the different aspects of Japan. Kyoto is the old
capital that has ancestral architecture worth seeing. The
current capital of Japan, Tokyo, is an urban jungle
especially the “electric” district of Asakusa. The town that I
will remember the most is the small coastal town of
Hitachi. This town had a nice view of the sea, ports, boats
and a few homes devastated by the tsunami of 2011. The
town also has a beautiful fish market with several seafood
restaurants. I loved the place because you could feel the
passion that the Japanese have for fishing and the
products of sea.
Despite the language barrier, I was well received in Japan. The Japanese are very welcoming
people and are always happy to share their customs and traditions. Japan is country with a rich
history and it was interesting to talk to people about their culture. You can feel their pride in their
values. If you can learn a few Japanese words and some of their customs, your effort will be
greatly appreciated. What surprised me the most was the number of time people were surprised
that I used chop-stick to eat and how often I was complimented on my dexterity with the chopsticks.
This international internship was the first time I worked in
an environment abroad. This gave me the opportunity to
develop both personally and professionally. Working
with a Japanese research team helped me learn to
adapt to new environments by integrating myself in a
new team that has different working norms and cultures.
Being able to adapt is an important research skill
because science is a field where collaboration,
exchange of ideas and networking is a must.
David Roy
The Fonds de recherche du Québec-Nature et technologies is acknowledged for his financial support of
this internship.
Info-CRIP No 6 – December 2012
11
CRIP 2011-2012 Graduates
Doctorat
Assaad Elias, 2011/11. Carboxyméthyl amidon et son complexe avec du chitosane comme
excipients pour des formulations pharmaceutiques; Director Mircea Alexandru Mateescu;
Collaborator Martin Lessard.
Chantal Forest, 2011/11. Étude fonctionnelle de l’opéron fimbriaire stg de Salmonella enterica
serovar Typhi; Director France Daigle.
Rima Habib, 2011/11. Rôle des régulateurs de fer pour la virulence chez Escherichia coli;
Director Charles M. Dozois.
Andrea Gomez Corredor, 2011/11. Caractérisation moléculaire et fonctionnelle de la protéine
REV du virus de l’immunodéficience bovine (VIB); Director Denis Archambault.
Richard Graveline, 2012/02. Étude des mécanismes moléculaires influençant la variation de
phase des adhésines P, F1651 et CS31A présentes chez des souches d’Escherichia coli
pathogènes; Directors Josée Harel and Christine Martin.
Marc Lemieux, 2012/06. Le carboxyméthyl amidon de faible à haut degré de substitution :
Excipient multifonctionnel pour des formes pharmaceutiques à administration orale; Director
Mircea Alexandru Mateescu and Patrick Gosselin.
Wilfried Saron, 2012/06. Expression chez les plantes de protéines recombinantes pour des
procédures vaccinales : Cas de l’artévirus porcin et de la flagelline de Salmonella typhimurium;
Director Denis Archambault and Fathey Sarhan.
Marie-Ève Charbonneau, 2012/08. Étude de la biogenèse de l’autotransporteur AIDA-1
d’Escherichia coli; Director Michael Mourez.
Sébastien Crépin, 2012/10. Role of the Pho regulon for virulence and gene regulation in
pathogenic E. coli; Director Charles M. Dozois and Josée Harel.
Master
Paul Kiswendsida Kaboré, 2012/02. Étude de prévalence et associations des gènes de
virulence et résistance aux antimicrobiens d’Escherichia coli de la flore intestinale du poulet
sain; Director John M. Fairbrother and Ann Letellier.
Claudia Hamida Syed, 2012/05. Escherichia coli STb toxin induces apoptosis in intestinal
epithelial cell lines; Director J. Daniel Dubreuil.
Info-CRIP No 6 – December 2012
12
New Initiatives
Since 2006, a total of 18 projects New Initiatives have been funded by CRIP. They allowed the
training of about 30 students and postdoctoral fellows and some research assistants. These
projects have generated scientific publications, presentation of conferences and posters to
national and international congresses, new research collaboration, new research proposals and
patent registration. And as these New Initiatives projects have an important impact on CRIP
research, they will maintain under our next 2013-2019 funding period.
The table below presents titles and investigators involved in New Initiatives projects in 2011 and
2012.
Investigator
Co-investigators
Project Titles
2012
La portion C-terminale de la protéine d’adhésion P97 de Mycoplasma
hyopneumoniae : Une molécule adjuvante à des fins vaccinales
Evaluation of a new treatment to control the exacerbated proinflammatory response in Streptococcus suis infected hosts
Rôle des biofilms bactériens dans la persistance de virus pathogènes
du porc dans l’environnement de la ferme
D. Archambault, UQAM
M. Segura, UdeM
C. A. Gagnon, UdeM
M. G. Gottschalk, UdeM
M. Segura, UdeM
M. Jacques, UdeM
C. A. Gagnon, UdeM
D. Grenier, U Laval
2011
Identification de facteurs de virulence chez le pathogène Streptococcus
suis à l’aide d’un modèle alternatif d’hôte
D. Grenier, U Laval
M. G. Gottschalk, UdeM
S. Charette, U Laval
Relation entre protozoaires aquatiques et survie d’Escherichia coli
producteur de Shiga toxines dans l’environnement : Implication du
régulon Pho
J. Harel, UdeM
F. Daigle, UdeM
C. M. Dozois, IAF, INRS
Results for the 2010 Competition – New initiatives
Zinc as an agent for the prevention of biofilm formation by pathogenic bacteria
Principal investigator:
Co-investigators:
Dr. Mario Jacques, FMV, Université de Montréal
Dr. Marie Archambault, FMV, UdeM
Dr. Daniel Grenier, Université Laval
Trainees:
Chan Wu, MSc student, FMV, UdeM
Dr. Yannick Tremblay, postdoctoral fellow, FMV, UdeM
Research Assistant:
Josée Labrie, FMV, UdeM
Biofilms are a structured community of micro-organisms enclosed in an extracellular matrix.
Biofilms are associated with the persistence of several infectious diseases. The extracellular
matrix of biofilms protects bacteria against the host immune system, antibiotics and
disinfectants. Recently, the laboratory of Dr Jacques demonstrated that zinc could inhibit the
formation of biofilms by Actinobacillus pleuropneumoniae, a bacterial pathogen of swine. Based
on this observation, the authors wanted to investigate the effect of zinc on the growth and biofilm
Info-CRIP No 6 – December 2012
13
formation of different swine pathogens including Bordetella bronchiseptica, Escherichia coli,
Haemophilus parasuis, Salmonella, Staphylococcus aureus et Streptococcus suis.
Bacteria were grown in 96-well plates under optimal biofilm formation conditions. The biofilms
were then stained with crystal violet. The presence of a biofilm was confirmed using confocal
laser scanning microscopy and the fluorescent stain FilmTracerTM FM® 1-43. At micromolar
concentrations, zinc weakly inhibited bacterial growth and blocked biofilm formation in a dosedependent manner for A. pleuropneumoniae, Salmonella Typhimurium and H. parasuis.
Additionally, biofilm formation by E. coli, S. aureus and S. suis was weakly inhibited by zinc.
"Confocal laser scanning microscopy images of Salmonella biofilms grown in the presence of different
ZnCl2 concentrations. Biofilms were stained with FilmTracer FM 1-43, a fluoresecent marker." Image A
indicates less presence of biofilm and image C indicates a strong biofilm formation.
Our results indicate that zinc has an inhibitory effect on biofilm formation of several bacterial
pathogens of porcine origin. However, the mechanism behind the antibiofilm properties of zinc
has yet to be characterized.
Structure determination of Streptococcus suis type 14 capsular polysaccharide
Principal investigator:
Co-investigators:
Dr. Mariela Segura, FMV, Université de Montréal
Dr. Marcelo Gottschalk, FMV, UdeM
Dr. Marie-Rose Van Calsteren, CRDA, AAC
Dr. Nahuel Fittipaldi, Methodist Hospital Research Institute, USA
Trainees:
David Roy, MSc student, FMV, UdeM
Guillaume Goyette-Desjardins, MSc student, FMV, UdeM
Cynthia Calzas, PhD student, FMV, UdeM
Streptococcus suis is a major swine pathogen responsible for important economic losses to the
swine industry worldwide and an emerging zoonotic agent of meningitis and streptococcal toxic
shock-like syndrome. In Southeast and East Asia, this bacterium affects not only people working
in the pig industry but also the general population, and it represents a significant public health
concern. In the very last years, as a direct consequence of intensified research efforts, large
amounts of data on putative virulence factors have appeared in the literature. Among them, the
capsular polysaccharide (CPS) is considered as the most critical for bacterial virulence. Of the
35 serotypes described based on CPS composition, most studies have been done on S. suis
serotype 2, which is classically associated with swine and human disease.
In addition to serotype 2, S. suis serotype 14 has been described as being an important swine
pathogen and an emerging zoonotic agent. Human cases of meningitis and severe sepsis with
Info-CRIP No 6 – December 2012
14
shock and multiple organ failure have been described in Western countries, including Canada,
and many strains are isolated each year from diseased pigs. Our knowledge on pathogenesis of
the disease induced by S. suis serotype 14 or on the virulence factors involved is scarce. The
complete genome sequence of this serotype has recently been published and we report here for
the first time the structure determination of S. suis serotype 14 CPS using chemical,
chromatographic, and spectroscopic methods. Lectin binding, physicochemical properties and
biosynthesis were also investigated. The CPS structure was compared with that of other
streptococcal antigens.
The CPS of S. suis serotype 14 was purified, chemically modified, and characterized. Sugar and
absolute configuration analyses gave the following CPS composition: D-Gal, 3; D-Glc, 1;
D-GlcNAc, 1; D-Neu5Ac, 1. The Sambucus nigra lectin, which recognizes the Neu5Ac(α2–
6)Gal/GalNAc sequence, showed binding to the native CPS. Sialic acid was found to be
terminal, and the CPS was quantitatively desialylated by mild acid hydrolysis. It was also
submitted to periodate oxidation followed by borohydride reduction and Smith degradation.
Sugar and methylation analyses, 1H and 13C nuclear magnetic resonance, and mass
spectrometry of the native CPS or of its specifically modified products allowed to determine the
repeating
unit
sequence:
[6)[Neu5Ac(α2–6)Gal(β1–4)GlcNAc(β1–3)]Gal(β1–
3)Gal(β1-4)Glc(β1]n. S. suis serotype 14 CPS has an identical sialic acid-containing side chain
as serotype 2 CPS, but differs by the absence of rhamnose in its composition. The same side
chain is also present in group B Streptococcus type Ia CPS, except that in the latter sialic acid is
2,3- rather than 2,6-linked to the following galactose. A correlation between the S. suis CPS
sequence and genes of the serotype 14 cps locus encoding putative glycosyltransferases and
polymerase responsible for the biosynthesis of the repeating unit is proposed.
Info-CRIP No 6 – December 2012
15
Advances in Research
Laboratoire d’épidémiologie et de médecine porcine (LEMP)
Sylvie D’Allaire, Benjamin Delisle
and Marie-Ève Lambert
Collaborators: Julie Arsenault and Martine
Denicourt, FMV, UdeM
Tools to improve PRRS control
Porcine reproductive and respiratory syndrome
(PRRS) costs the Canadian swine industry approximately 130 Million CAD annually. Increasing
our knowledge of PRRS virus (PRRSV) epidemiology is one of the best investments to improve
disease management in the field and to reduce production costs. Rapid advancement in
molecular biology as led to the development of molecular diagnosis tools that are readily
available to the practitioners. The challenge is now to integrate the molecular-based data with
traditional epidemiological data to gain a maximum knowledge on PRRSV transmission to
facilitate herd and regional control of the disease.
The LEMP continuously updates and maintains a Quebec PRRSV database of more than 2600
sequences, collected over the past 15 years, combined with geographical and demographical
herd information, serving both field and research purposes. Through this database, the
laboratory provides active and constant support to participating veterinarians for several Quebec
ARC&E initiatives. Reports on circulating PRRSV strains are produced using phylogenetic,
spatial and statistical software to facilitate the interpretation of sequencing results and to
maximize its use in the field. Also, members of the LEMP currently investigate various reporting
and analysis tools integrating sequence data for area regional control and elimination (ARC&E)
projects in Canada in order to propose optimal methods of reporting circulating strains in support
to these initiatives. This project aims to compile existing reporting methodologies and outputs
available, to describe and prioritize the applications of PRRSV sequencing data in ARC&E and
to evaluate current and potential analysis options to meet identified priorities. The value,
limitations, and ongoing feasibility of delivering these reports on a timely basis to current and
future projects will be presented as well as the objectives and feasibility of comparing sequence
data between different ARC&E projects.
From a research perspective, several direct and indirect transmission pathways of the virus have
been reported, but their relative importance remains to be established. The LEMP is currently
developing a methodology integrating traditional and molecular data in order to assess various
aspects of PRRSV transmission dynamic using a multidisciplinary approach. More specifically,
several methods available for classifying PRRSV strains into genetic clusters are and will be
evaluated as well as the potential for generating automatically the classification to manage more
efficiently incoming PRRSV sequence submissions on a timely basis. Recombination events will
also be examined in order to avoid phylogenetic misclassification of strains, to gain insight into
molecular structures involved in sequence cross over and to describe herd and neighbourhood
characteristics of farms involved in these events. Spatial and temporal dispersal of PRRSV
strains will be evaluated to understand dynamic of PRRSV populations as a part of a global
assessment of the importance of various sources of contamination.
Members of the LEMP are also interested in developing a methodological approach to identify
the most likely source of virus introduction into a herd by analyzing the likelihood of herd
Info-CRIP No 6 – December 2012
16
contacts through various pathways (e.g. animal source, employees, airborne transmission…)
among herds sharing genetically similar PRRSV strains. This could serve as guidelines to
prioritize interventions on specific risk factors to limit introduction of a new PRRSV strain taking
into account the different herd characteristics such as production type, flow of animals, etc. All
the information gained through these latter research projects will be helpful in decision making
and risk assessment.
For further information, please contact:
Dr. Sylvie D’Allaire, sylvie.dallaire@umontreal.ca
Benjamin Delisle, benjamin.delisle@umontreal.ca
Dr. Marie-Ève Lambert, marie-eve.lambert@umontreal.ca
Following is a list of LEMP articles published during the last year.
Epidemiological investigations in regard to porcine reproductive and respiratory
syndrome (PRRS) in Quebec, Canada. Part 1: Biosecurity practices and their
geographical distribution in two areas of different swine density
Lambert ME, Poljak Z, Arsenault J, D’Allaire S
Prev Vet Med. 2012 April; 104 (1-2): 74-83.
Porcine reproductive and respiratory syndrome virus (PRRSV) is a considerable threat to the
swine industry and implementing biosecurity measures is essential for the control of its
transmission. The aims of this study were: (1) to describe biosecurity practices in production
sites located in a moderate density (MD) and a high density (HD) pig area according to
production type; (2) to group sites in different patterns according to their biosecurity practices;
and (3) to determine the geographical distribution of sites according to biosecurity patterns.
Biosecurity practices were selected based on PRRS epidemiology. A questionnaire was
completed on 125 breeding sites (MD = 54; HD = 71) and 120 growing (HD) sites, between 2005
and 2008. Depending on area and production type, the frequency of biosecurity practices used
ranged from 0 to 2% for barrier at site entrance, 0 to 19% for use of shower, 25 to 35% for
washing truck between loads of pigs, 51 to 57% for absence of rendering or rendering without
access to the site, and 26 to 51% for absence of gilt purchase or purchase with quarantine.
Better practices pertaining to entrance protocol (i.e. “no-entry” sign, shower, ≥24 h downtime)
were reported more frequently on breeding sites in the MD than the HD area (P < 0.05). In the
HD area, growing sites had in general a lower level of biosecurity than breeding sites. Using a
two-step clustering procedure performed separately for breeding and growing sites, two different
patterns were obtained for each production type, which corresponded to a high and low level of
biosecurity. For breeding sites, a higher biosecurity level was observed at sites located away
from other pig sites, set at more than 300 m from the public road, having higher sow inventory,
or being part of an integrated production (P < 0.05). Spatial clusters of sites for each biosecurity
pattern were detected. This study identified some shortcomings regarding biosecurity that should
be addressed before implementing any PRRSV regional control. Vicinity of sites with different
biosecurity levels also suggests difficulties in planning priorities of intervention based on
geographical distribution of sites.
Info-CRIP No 6 – December 2012
17
Epidemiological investigations in regard to porcine reproductive and respiratory
syndrome (PRRS) in Quebec, Canada. Part 2: Prevalence and risk factors in breeding
sites
Lambert ME, Arsenault J, Poljak Z, D’Allaire S
Prev Vet Med. 2012 April; 104 (1-2): 84-93.
Porcine reproductive and respiratory syndrome virus (PRRSV) is a major threat for swine
industry and understanding factors involved in its epidemiology is undoubtedly essential for
disease control. As a part of larger project, a cross-sectional study was performed on breeding
sites in a moderate density area of swine production in Quebec to estimate the prevalence of
PRRSV infected sites and to evaluate if characteristics of sites and biosecurity practices, either
as specific measures or as a global score, were associated with PRRSVstatus. A questionnaire
and diagnostic procedures were performed on 54 breeding sites between September 2006 and
August 2008. A biosecurity score that had been previously computed using two-step clustering
procedure was used, classifying breeding sites into two biosecurity patterns (high vs. low)
according to 21 specific biosecurity measures. The apparent prevalence of PRRSV infected
sites was 74.0% (95% CI, 60.3–85.0). Univariable and multivariable logistic regression models
with robust standard errors adjusting for potential clustering of sites due to same ownership were
computed. In a first multivariable model evaluating characteristics of sites and specific
biosecurity variables, four main effects were significantly associated (P < 0.05) with PRRSV
positive status: large pig inventory (OR: 10.7), proximity to closest pig site (OR: 7.3), absence of
shower (OR: 8.7) and free access to the main entrance of the site by the rendering truck (OR:
7.0). In a second multivariable model including a global biosecurity score as a surrogate for a
specific pattern of biosecurity measures, this score was not retained in the final model. The
adjusted population attributable fractions were 16% for the proximity to closest pig site variable,
27% for the absence of shower variable, and 10% for the free access to main entrance of the
site by the rendering truck. These two latter biosecurity measures, manageable directly on the
site, should be prioritized and be part of any intervention strategy designed for PRRSV control.
Correlation among genetic, Euclidean, temporal, and herd ownership distances of
porcine reproductive and respiratory syndrome virus strains in Quebec, Canada
Lambert ME, Arsenault J, Poljak Z, D’Allaire S
BMC Veterinary Research 2012 June, 8:76.
Porcine reproductive and respiratory syndrome (PRRS) is a viral disease that has a major
economic impact for the swine industry. Its control is mostly directed towards preventing its
spread which requires a better understanding of the mechanisms of transmission of the virus
between herds. The objectives of this study were to describe the genetic diversity and to assess
the correlation among genetic, Euclidean and temporal distances and ownership to better
understand pathways of transmission. A cross-sectional study was conducted on sites located in
a high density area of swine production in Quebec. Geographical coordinates
(longitude/latitude), date of submission and ownership were obtained for each site. ORF5
sequencing was attempted on PRRSV positive sites. Proportion of pairwise combinations of
strains having ≥98% genetic homology were analysed according to Euclidean distances and
ownership. Correlations between genetic, Euclidean and temporal distances and ownership
were assessed using Mantel tests on continuous and binary matrices. Sensitivity of the
correlations between genetic and Euclidean as well as temporal distances was evaluated for
different Euclidean and temporal distance thresholds. An ORF5 sequence was identified for 132
of the 176 (75%) PRRSV positive sites; 122 were wild-type strains. The mean (min-max)
genetic, Euclidean and temporal pairwise distances were 11.6% (0–18.7), 15.0 km (0.04-45.7)
and 218 days (0–852), respectively. Significant positive correlations were observed between
genetic and ownership, genetic and Euclidean and between genetic and temporal binary
Info-CRIP No 6 – December 2012
18
distances. The relationship between genetic and ownership suggests either common sources of
animals or semen, employees, technical services or vehicles, whereas that between genetic and
Euclidean binary distances is compatible with area spread of the virus. The latter correlation was
observed only up to 5 km. This study suggests that transmission of PRRSV is likely to occur
between sites belonging to the same owner or through area spread within a 5 km distance.
Both should be considered in the perspective of prevention.
Porcine reproductive and respiratory syndrome virus diversity of Eastern Canada swine
herds in a large sequence dataset reveals two hypervariable regions under positive
selection
Delisle B, Gagnon CA, Lambert ME, D’Allaire S
Infect Genet Evol. 2012 Jul;12(5):1111-1119.
Porcine reproductive and respiratory syndrome virus (PRRSV) is known to be genetically highly
variable, but knowledge of sequence diversity from Eastern Canada and its degree of genetic
plasticity in or near the principal neutralizing epitope (PNE) in association with evolutionary
selective pressure is limited. The purposes of our study were to investigate the extent of strain
diversity, the existing glycotypes and the amino acid sites under selective evolutionary pressure
in its encoded protein, GP5, for a dataset of 1301 sequences (1998 to 2009). This was
addressed by partitioning and clustering into subgenotypes a large number of open reading
frame 5 sequences from the province of Quebec and analyzing the content of these
subgenotypes. The overall pairwise diversity was 12% and was comparable to what has been
reported around the world. The mean diversity for sequences within subgenotypes was around
7%. No marked variations in subgenotype emergence could be observed through time. Thirtyeight GP5 glycotype patterns were observed which included a newly identified site at position
N57 which was already present in 1998. These patterns possessed one to six N-glycosylation
sites in total and could be located in eight different positions. No obvious grouping of glycotypes
could be established in relation to subgenotypes. Positions N44 and N51 were confirmed to be
fixed N-glycosylation positions, whereas other positions where found to be shifting and located
in or near hypervariable regions (HVR) 1 and 2. Both HVR were under selective evolutionary
pressure in half of all subgenotypes including vaccine-like groups. Conversely, the PNE flanked
by both HVR was well conserved amongst most subgenotypes demonstrating potential
molecular constraint in a probable viral binding region. The analysis of this dataset increased
knowledge of evolutionary change inferred from genetic data, more specifically regarding the
implications of both HVRs in PRRSV diversity.
Gilt replacement strategies used in two swine production areas in Quebec in regard to
porcine reproductive and respiratory syndrome virus
Lambert ME, Denicourt M, Poljak Z, D’Allaire S
J Swine Health Prod. 2012;20(5):223–230.
The objectives of this study was to describe gilt replacement strategies in regard to porcine
reproductive and respiratory syndrome virus (PRRSV) and to assess differences between high
density (HD) and moderate density (MD) pig areas. A cross-sectional study was conducted in
breeding sites located in an HD (n = 68) and an MD area (n = 52) in Quebec between May 2005
and August 2008. A questionnaire on strategies used to introduce replacement gilts was
completed and PRRSV status was assessed by enzyme-linked immunosorbent assay or
reverse-transcription polymerase chain reaction. Sites housing at least one pig positive by either
test were classified as PRRSV-positive. Strategies were described according to herd
characteristics, PRRSV status, and area. Self-replacement and purchase of mature or immature
gilts were observed on 37%, 35%, and 28% of sites, respectively. In positive sites purchasing
mature gilts, 18% had a PRRSV-positive supplier, and gilts were introduced either directly into
Info-CRIP No 6 – December 2012
19
the sow herd (15%) or after isolation (41%) or acclimatization (44%). Most positive sites
purchasing immature gilts practiced acclimatization (93%), either by commingling gilts with
commercial pigs (93%) or inoculating serum (7%). Acclimatization processes were rarely
monitored through diagnostic procedures. Lower sow inventory, higher prevalence of PRRSV
infection, and higher frequency of self-replacement were observed in the HD compared to the
MD area. Negative and positive sites practicing voluntary exposure to PRRSV both clustered
spatially within the MD area. Replacement strategies may have weaknesses that should be
addressed to facilitate PRRSV management at the herd and regional levels.
Alopecia areata and humpy-back syndrome in suckling piglets
Drolet R, Denicourt M, D’Allaire S
Can Vet J. 2012; 53: 865-869.
This report describes in several nursing piglets an uncommon variant of humpy-back syndrome
associated with multiple rib fractures and multisystemic vasculitis and for the first time in swine a
skin disease consistent with alopecia areata. Both conditions were observed concurrently on the
farm and even in some piglets. Possible causes are discussed.
2013 CRIPA Scientific Communications
We invite you to participate to our next scientific communications for 2013! Place these
important dates on your agenda and keep an eye on the CRIPA Bulletin for further
information and registration.
May 8th
CRIPA Scientific Colloquium organized in the frame of the "81ième congrès de l’Acfas" at
Université Laval, Québec: "Le microbiote animal : Une question d’équilibre!"
May 9th
CRIPA 6th Annual Symposium
We thank Yannick Tremblay and Jenny-Lee Thomassin for the translation of the Info-CRIP 6.
Info-CRIP No 6 – December 2012
20