7th Poultry Days - Uniwersytet Rolniczy w Krakowie
Transcription
7th Poultry Days - Uniwersytet Rolniczy w Krakowie
University of Agriculture in Krakow Faculty of Animal Sciences International Conference on Biotechnology and Welfare in Animal Science with a session on „7th Poultry Days” Krakow • 23–24 June 2016 Editor-in-Chief Prof. dr hab. inż. Józef Bieniek Scientific Committee Chairman Prof. dr hab. Czesław Klocek Dean of Faculty of Animal Sciences, University of Agriculture in Krakow Members: Prof. Arieh Gertler, The Hebrew University of Jerusalem, Israel Prof. dr hab. Eugeniusz Herbut, National Research Institute of Animal Production in Krakow, Poland Prof. dr hab. Roman Kołacz, Wroclaw University of Environmental and Life Sciences, Poland Prof. Jerzy Niedziółka, University of Agriculture in Krakow, Poland Prof. Marek Spinka, Institute of Animal Science in Prague, Czech Republic Prof. dr hab. Dorota A. Zięba, University of Agriculture in Krakow, Poland Organizing Committee Chairman Dr hab. Piotr Micek, prof. UR Vice Dean for Organisation and Development of Faculty of Animal Sciences, University of Agriculture in Krakow Members: Dr hab. inż. Marcin Lis (Secretary), University of Agriculture in Krakow, Poland Dr Krzysztof Andres, University of Agriculture in Krakow, Poland Dr inż. Jadwiga Flaga, University of Agriculture in Krakow, Poland Dr inż. Małgorzata Gumułka, University of Agriculture in Krakow, Poland Mgr Beata Majka, University of Agriculture in Krakow, Poland Dr inż. Katarzyna Kirsz, University of Agriculture in Krakow, Poland Dr inż. Stanisław Łapiński, University of Agriculture in Krakow, Poland Dr inż. Ewa Ocłoń, University of Agriculture in Krakow, Poland Dr hab. Krzysztof Pawlak, University of Agriculture in Krakow, Poland The organizers are not responsible for the content of reports Pre-press preparation Publishing House of the University of Agriculture in Krakow Cover design Katarzyna Kirsz ISBN 978-83-64758-38-6 Publishing House of the University of Agriculture in Krakow 31-425 Kraków, al. 29 Listopada 46 tel. (12) 662 51 57, 662 51 59 e-mail: wydawnictwo@ur.krakow.pl www.wydawnictwo.ur.krakow.pl Patronat Honorowy Krzysztof Jurgiel – Minister Rolnictwa i Rozwoju Wsi Jacek Krupa – Marszałek Województwa Małopolskiego prof. dr hab. Włodzimierz Sady – Rektor Uniwersytetu Rolniczego w Krakowie Partnerzy Konferencji Miasto Kraków Polski Oddział Światowego Stowarzyszenia Wiedzy Drobiarskiej Krajowa Rada Drobiarstwa – Izba Gospodarcza w Warszawie Patronat Medialny Mecenat Sponsorzy Table of contents Plenary papers............................................................................................................................................................................... 13 Invited lectures Arieh Gertler Resistin/adiponectin/FGF21/palmitic acid interplay in SH-SY5Y neural cells and DIO rodents: a novel mechanism contributing to insulin resistance...................................................................................................... 15 Roman Kołacz Aktualne problemy dobrostanu drobiu • Current problems of poultry welfare............................................................ 16 Andrzej Sechman, Piotr A. Antos, Dorota Katarzyńska, Anna Hrabia Chicken ovary – a place of dioxin and polychlorinated biphenyl detoxification .......................................................... 21 Session 1. Animal welfare and product quality ....................................................................................................................... 23 Plenary lectures Piotr SJ Sowinski Roles, tasks and responsibilities of the Designated Veterinarian under Directive 2010/63/EU.................................. 25 Paweł Paraponiak Fatty acid profile of lamb meat from highly productive breeds of sheep reared under organic conditions .............. 27 Presentations Victoria I. Borodulina, N.A. Sadomov The influence of mycotoxin’s adsorbent of new generation “Funginorm” on the productivity of fattening pigs...... 28 Veronika Čoudková, J. Starý, Miroslav Maršálek Mycotoxins in dairy cows diet in south-west Bohemia .................................................................................................... 30 Bogusława Długosz, Romana Augustyn, Magdalena Pieszka, Jarosław Łuszczyński, Tomasz Próchniak, Barbara Jaklińska, Karina Świderska, Zenon Podstawski, Monika Stefaniuk-Szmukier, Weronika Pisarczyk Selected reproductive indicators for mares of the Hucul Pony breed ............................................................................. 31 Ewa Drąg-Kozak, Ewa Łuszczek-Trojnar, Magdalena Socha, Grzegorz Gosiewski, Włodzimierz Popek Bioaccumulation and elimination of cadmium in tissues of Prussian carp (Carassius gibelio B.) under controlled conditions............................................................................................................................................................. 33 Jan Dybała, Aleksandra Milewska, Joanna Wiśniewska, Aleksandra Cebulska, Wojciech Kapelański The analysis of welfare Visayan warty pig (Sus cebifrons) kept in selected zoos............................................................ 34 Kinga Fic, Aleksandra Woszczenko, Agnieszka Krzystanek, Natalia Sowińska The influence of frequency of enrichments replacement on behavior of laboratory guinea pigs ............................... 35 Iwona Guja, Stanisław Łapiński, Piotr Niedbała Is the fur chewing in animals caused by Slitrk1 gene mutation? ..................................................................................... 36 6 Andrzej Kaczor, Urszula Kaczor Effect of feeding technology on calf behaviour .................................................................................................................. 37 Andrzej Kaczor, Urszula Kaczor Effect of watering of cows in the milking parlour on their welfare ................................................................................. 38 Aldona Kawęcka, Michał Puchała, Anna Miksza-Cybulska Effect of slaughter age on physicochemical parameters of lamb meat............................................................................ 39 Aldona Kawęcka, Jacek Sikora Effect of housing system on slaughter traits of native Świniarka lambs ......................................................................... 40 Aldona Kawęcka, Ewa Sosin-Bzducha Meat quality in lambs of native breed fed the diet supplemented with maize DDGS .................................................. 41 Agata Kokocińska The impact of environmental enrichment on exploratory and hunting behavior of laboratory and companion animals .................................................................................................................................................................................... 42 Stanisław Łapiński, Iwona Guja, Katarzyna Mrozicka, Piotr Niedbała, Viktor A. Ilyukha Behaviour of Chinchilla lanigera male kept under farm conditions – preliminary studies ......................................... 44 Jarosław Łuszczyński, Kinga Dobrzańska, Magdalena Pieszka, Weronika Pisarczyk, Bogusława Długosz, Romana Augustyn, Zenon Podstawski, Monika Stefaniuk-Szmukier The differences in the biometric dimensions of the Silesian horses depending on the share of other breeds blood in their pedigrees ....................................................................................................................................................... 45 Elena L. Mikulich Influence of timely gutting fish on intensity of infection of muscle tissue by the Anisakis larvae ............................. 47 Małgorzata Natonek-Wiśniewska, Piotr Krzyścin The use of PCR and real-time PCR for qualitative and quantitative determination of horse meat ............................ 48 Patryk Nowacki, Ewa Łuszczek-Trojnar, Ewa Drąg-Kozak, Magdalena Socha, Włodzimierz Popek Concentration of heavy metals in increments of common carp’s scales (Cyprinus carpio L.) .................................... 49 Agnieszka Otwinowska-Mindur, Ewa Ptak The freezing point of milk of Polish Holstein-Friesian cows ........................................................................................... 51 Paweł Paraponiak Concentration of harmful gas components in organic sheep houses according to the season ................................... 52 Paweł Paraponiak Variation in lamb behaviour according to the housing system ....................................................................................... 53 Magdalena Pieszka, Anna Halek, Jarosław Łuszczyński, Bogusława Długosz, Romana Augustyn, Monika Stefaniuk-Szmukier, Zenon Podstawski, Weronika Pisarczyk Characterisation of the population of border collie dogs breed in Poland .................................................................... 54 Zenon Podstawski, Justyna Kądzior, Bogusława Długosz, Romana Augustyn, Monika Stefaniuk-Szmukier, Magdalena Pieszka, Jarosław Łuszczyński, Weronika Pisarczyk Characteristics of work of chosen hippotherapy centres in southern Poland .............................................................. 56 Jan B. Pyś, Zygmunt M. Kowalski, Marta Malkiewicz, Jadwiga Flaga, Łukasz Korytkowski, Vanessa Demey The effect of bacteria addition on the chemical and microbiological composition of the whole plant corn silage ............................................................................................................................................................................... 58 Jan B. Pyś, Zygmunt M. Kowalski, Marta Malkiewicz, Jadwiga Flaga, Łukasz Korytkowski, Vanessa Demey The effect of bacteria addition on the fermentation quality and aerobic stability of the whole plant corn silage ............................................................................................................................................................................... 60 Iwona Radkowska Changes in some fat parameters during storage of butter made from the milk of cows under two feeding systems..................................................................................................................................................................................... 61 Iwona Radkowska, Adam Radkowski Vitamin and cholesterol content of milk depending on type of feeding ........................................................................ 62 Adam Radkowski, Iwona Radkowska Effect of foliar fertilization with silicon on quality and nutritive value of meadow sward silages ............................. 63 Adam Radkowski, Iwona Radkowska Quality and nutritive value of silages made from selected grass-legume mixtures ...................................................... 64 Alicja Satoła, Ewa Ptak, Wojciech Jagusiak Genetic relationship between conformation traits and lactose percentage and urea concentration in milk of Polish Holstein-Friesian cows .......................................................................................................................................... 65 Ewa Sosin-Bzducha, Michał Puchała Effect of breed and aging time on physicochemical and organoleptic quality of beef.................................................. 66 Petr Tejml, Miloslav Šoch Effect of litter size on mortality of new born cavies’ youngs ............................................................................................ 68 Iwona Tomczyk-Wrona Effect of herd size on implementation of the genetic resources conservation programme for Hucul horses ........... 69 Iwona Tomczyk-Wrona Effect of implementing the genetic resources conservation programme on the distribution of Hucul horses in Poland ................................................................................................................................................................................. 71 Michał Tracz, Agnieszka Jackowska-Tracz, Krzysztof Anusz Polish collection and communication of inspection results system and food chain information system as tools for welfare improvement at slaughterhouse ......................................................................................................... 73 Dorota Wojtysiak, Magdalena Górska, Joanna Wojciechowska Effect of ageing time on microstructure and tenderness of pig semimembranosus muscle........................................ 74 Justyna Żychlińska-Buczek, Edyta Bauer, Andrzej Ochrem, Elżbieta Sowula-Skrzyńska, Grzegorz Skrzyński The effect of Pedi bolus specimen on the improvement of cows’ hooves health ........................................................... 75 Session 2. Biotechnology and animal models.......................................................................................................................... 77 Plenary lectures Viktor A. Ilyukha, Svetlana N. Sergina, Ekaterina P. Antonova, Evgeny A. Khizhkin, Lyudmila B. Uzenbaeva, Aleksandra G. Kizhina, Tatiana N. Ilyina, Irina V. Baishnikova, Stanisław Łapiński, Piotr Niedbała, Marcin W. Lis, Oleg V. Trapezov, Yongping Xu Fur-bearing animals as the models in biomedical research ............................................................................................. 79 Alicja Jozkowicz Transgenic mice in cancer research ..................................................................................................................................... 80 Presentations Romana Augustyn, A. Pustuła, Bogusława Długosz, Tomasz Próchniak, Magdalena Pieszka, Jarosław Łuszczyński, Weronika Pisarczyk, Monika Stefaniuk-Szmukier, Zenon Podstawski Influence endurance workout on selected physiological parameters of Arabian horses ............................................. 81 Agnieszka Baranowska, Beata Skowron, Katarzyna Ciesielczyk, Justyna Domagała, Piotr J. Thor Experimental gender related obesity effect of diet ............................................................................................................ 83 Aneta Bocheńska, Agnieszka Pietsch-Fulbiszewska, Natalia Sowińska, Katarzyna Kirsz Method of positioning the catheter into cisterna magna and fourth ventricle of the brain in sheep ......................... 84 Taras Chоkаn, Anna Radko, Serhiy Tarasjuk, Agnieszka Szumiec, Dominika Rubiś Intrаbreed variability of the Ukrainian Mountain Carpathian sheep genetic structure under using of the different types of molecular genetic markers .......................................................................................................... 85 Jarosław Chyb, Gabriela Cora, Piotr Lisowski, Magdalena Socha, Mirosława Sokołowska-Mikołajczyk The influence of orexin B on spontaneous and GnRH-stimulated LH secretion from dispersed pituitary cells of female Prussian carp................................................................................................................................................. 86 Iwona Dobosz, Katarzyna Knapczyk-Stwora Prenatal exposure to anti-androgen alters expression of transcription factor forkhead L2 (Foxl2) in the fetal porcine ovaries .................................................................................................................................................... 87 7 Ewelina Górowska-Wójtowicz, Laura Pardyak, Alicja Kamińska, Anna Hejmej, Małgorzata Kotula-Balak, Joanna Dulińska-Litewka, Barbara Bilińska Does 2-hydroxyflutamide affect localization of kinases involved in non-classical testosterone pathway in prostate cancer cell line LNCaP? ..................................................................................................................................... 88 Gabriela Halik, Andrzej Łozicki, Agata Koziorzębska, Mateusz Makarski, Magdalena Matusiewicz The impact of the ration supplemented with synthetic b-carotene and silage with a pumpkin as a source of natural b-carotene on the bioactive compounds in colostrum ................................................................................... 89 Alicja Kamińska, Laura Pardyak, Ewelina Górowska-Wójtowicz, Małgorzata Kotula-Balak, Barbara Bilińska, Katarzyna Szczęśniak, Anna Hejmej Effect of phosphodiesterase 8B ablation on mouse spermatogenesis and steroidogenesis .......................................... 90 Tatiana N. Ilyina, Irina V. Baishnikova, Viktor A. Ilyukha, Svetlana N. Sergina, Stanisław Łapiński Comparison of antioxidant status in farmed raccoon dog from Poland and Russia .................................................... 91 Katarzyna Kirsz, Małgorzata Szczęsna, Dorota A. Zięba Agonist and antagonist of 5-HT2B/2C receptor and ghrelin activate intracellular signal cascades responsible for melatonin biosynthesis in the ovine pineal gland ....................................................................................................... 92 Urszula Kaczor, Magdalena Luber, Mirosław Kucharski, Andrzej Kaczor T FOXO3 genetic variability in selected Polish sheep breeds .......................................................................................... 93 Mirosław Kucharski, Urszula Kaczor, Katarzyna Piórkowska, Karolina Pastuszak Identification of SNP c.*1116A > G and c.2218G > A in the SCD gene of sheep breeds raised in Poland................ 94 Mirosław Kucharski, Urszula Kaczor, Katarzyna Piórkowska, Katarzyna Zemsta Identification of SNP c.391 – 66C > T and c.529 + 27G > C in the PPARγ gene of sheep breeds raised in Poland ................................................................................................................................................................................. 95 Marta Kuchta-Gładysz, Katarzyna Andraszek, Piotr Niedbała, Agnieszka Otwinowska-Mindur, Anna Grzesiakowska, Olga Szeleszczuk Analysis of the structure of sperm chromatinin breeding foxes...................................................................................... 96 Magdalena Mika, Agnieszka Wikiera, Anna Antończyk, Anna Starzyńska-Janiszewska Effect of butylated hydroxytoluene on bioavailability of dietary nutritional compounds ........................................... 97 Edyta Molik, Michał Błasiak, Tomasz Misztal, Katarzyna Romanowicz, Dorota A. Zięba The role of light and exogenous melatonin in the regulation of prolactin secretion in sheep..................................... 98 Agnieszka Mostek, Mariola A. Dietrich, Błażej Westfalewicz, G. Hołownia, Andrzej Ciereszko Cryopreservation of bull semen produces an oxidative damage of sperm proteins ..................................................... 99 Maria Motal, Kamil Wartalski, Małgorzata Duda The pre-established reserve of porcine preantral follicles – advanced method of enzymatic isolation and identification ........................................................................................................................................................................... 101 Klaudia Nalik, Kinga Fic, Patryk Hartwich, Paweł Stręk, Michał Nowakowski Usefullness of disected sheep heads in paranasal sinuses endoscopy for physicians.................................................... 102 Piotr Niedbała, Gabriela Lipień, Olga Szeleszczuk, Marta Kuchta-Gładysz, Stanisław Łapiński, Iwona Guja Preliminary research of chosen biochemical compounds’ level in the semen of breeding chinchillas (Chinchilla lanigera)............................................................................................................................................................... 103 Marta Pasternak Analysis of KM1 allele frequency in intron 13 of the KIT gene in solid-coloured Hucul horses with white markings .............................................................................................................................................................. 104 Magdalena Socha, Jarosław Chyb, Mirosława Sokołowska-Mikołajczyk, Ewa Drąg-Kozak, Włodzimierz Popek In vitro LH secretion by female common carp (Cyprinus carpio L.) pituitary cells incubated with popular herbicide Roundup ................................................................................................................................................................ 105 Grzegorz Sokołowski, Ricardo Faundez, Zuzanna Sandowska-Markiewicz, Marta Gajewska, Anna Niwińska, Anita Tyl-Bielicka, Marek Woszczyński, Katarzyna Kermen-Germaniuk, Sebastian Dąbrowski, Monika Petrajtis-Golobow, Jerzy Ostrowski Application of time-lapse monitoring of embryo development (TLMED) in different mouse strains ..................... 106 8 Natalia Sowińska, Anna Filipczyk, Katarzyna Frankowska, Klaudia Nalik, Kinga Fic The role of cumulus cells in in vitro maturation and in vitro fertilization of oocytes of domestic cat ....................... 107 Monika Stefaniuk-Szmukier, Katarzyna Ropka-Molik, Katarzyna Piórkowska, Jarosław Łuszczyński, Magdalena Pieszka, Tomasz Szmotała, Bogusława Długosz, Romana Augustyn, Weronika Pisarczyk, Zenon Podstawski Genetic variability in equine GDF9 in Arab and Thoroughbred mares ......................................................................... 108 Małgorzata Szczęsna, Katarzyna Kirsz, Dorota A. Zięba Pregnancy-induced changes of leptin concentration in Polish Longwool ewes............................................................ 109 Magdalena Szyndler-Nędza, Katarzyna Ropka-Molik, Katarzyna Piórkowska, Aurelia Mucha, Marek Babicz, Tadeusz Blicharski Analysis of changes in the frequency of FST genotypes depending on inbreeding of native Puławska sows.......................................................................................................................................................................................... 110 Aleksey R. Unzhakov, Nikolay N. Tyutyunnik, Svetlana N. Sergina, Nadezhda P. Chernobrovkina, Ekaterina P. Antonova, Lyudmila B. Uzenbaeva, Irina V. Baishnikova, Viktor A. Ilyukha, Stanisław Łapiński, Piotr Niedbała Extract from pine needle enriched with L-arginine prevents the protein malnutrition of American mink (Neovison vison) kits .............................................................................................................................................................. 112 Vasyl Vlizlo, Natalia Kuzmina, Dmytro Ostapiv, Mykola Sharan, Iryna Yaremchuk, Julia Bodnar Glutathione peroxidase activity, respiration intensity and spermatozoa survival in bull ejaculates .......................... 113 Kamil Wartalski, Gabriela Gorczyca, Marta Hereta, Małgorzata Duda The evaluation of the ability to differentiate of putative stem cells from mature pig ovaries ...................................... 114 Jerzy Wiater, Janusz Karasiński Characterization and possible involvement of mesenchymal stem cells in endometrium of porcine uterus model ....................................................................................................................................................................................... 115 Jerzy Wiater, Janusz Karasiński, Jacek Jura, Zdzisław Smorąg, Ryszard Słomski, Marek Romek Pig with reduced expression of the Galα(1,3)Gal epitope as a potential donor of skin grafts – preliminary study......................................................................................................................................................................................... 116 Joanna Zubel-Łojek, Anna Latacz, Ewa Ocłoń The changes of CRP and IL-10 secretion from piglet lymphocytes in response to adiponectin ................................. 117 Session 3. ”7th Poultry Days” – current problems of poultry and ornamental bird breeding ......................................... 119 Plenary lectures Veronika Pistová, Henrieta Arpášová, Cyril Hrnčár The effect of the humic acid and herbal additive supplement on production and carcass parameters of broiler chicken .................................................................................................................................................................. 121 Maria Siwek, Anna Sławińska, Katarzyna Stadnicka, Aleksandra Dunisławska, Arkadiusz Płowiec, Giuseppe Maiorano, Marek Bednarczyk In ovo technology – efficient method of chicken microbiome stimulation ................................................................... 122 Presentations Michalina Adaszyńska-Skwirzyńska, Danuta Szczerbińska Effect of lavender essential oil (Lavandula angustifolia) on production indicators of broiler chicken ..................... 123 Krisztián Balogh, Nikolett Tokodi, Erika Zándoki, Judit Szabó-Fodor, Csilla Pelyhe, Andrea Bócsai, Miklós Mézes Effects of fumonisin contaminated diet on glutathione redox system and lipid peroxidation processes in broiler chickens ................................................................................................................................................................. 125 Mateusz Bucław Seasonal changes in the behavior of emu (Dromaius novaehollandiae) under farm conditions ................................ 126 Jolanta Calik The content of selected chemical components in hen eggs depending on egg production cycle .............................. 128 9 Jolanta Calik, Katarzyna Połtowicz, Robert Gąsior Effect of caponizing Yellowleg Partridge (Ż-33) cockerels on meat quality .................................................................. 129 Jolanta Calik, Michał Puchała, Joanna Obrzut Evaluation of productivity and egg quality in Sussex (S-66) laying hens during the first two years of production ......................................................................................................................................................................... 130 Magdalena Dykiel, Zofia Sokołowicz The welfare of hens in various systems of extensive farming .......................................................................................... 131 Małgorzata Dżugan, Monika Wesołowska, Marcin Lis, Teodora Król, Jakub Nowak, Jerzy W. Niedziółka In ovo study on calcium protective effect against cadmium embryotoxicity ................................................................ 132 Csaba Fernye, Márta Erdélyi, Krisztián Balogh, Zsolt Ancsin, Andrea Bócsai, Miklós Mézes Preliminary study on some parameters of meat quality and antioxidant defence of grey partridge (Perdix perdix) ....................................................................................................................................................................... 133 Marian Foltyn, Vojtěch Rada, Martina Lichovníková The infuence of rapeseed on growth performance in broiler diets ................................................................................ 134 Karolina Goździewska-Harłajczuk, Joanna Klećkowska-Nawrot, Karolina Barszcz The microscopic study of the conjunctiva-associated lymphoid tissue (CALT) in Indian peafowl (Pavo cristatus) ...................................................................................................................................................................... 135 Magdalena Graczyk, Krzysztof Andres, Ewa Kapkowska, Tomasz Szwaczkowski Monitoring of the genetic diversity of the Zatorska goose based on the pedigree information ................................ 136 Emília Hanusová, Anton Hanus, Cyril Hrnčár, Marta Oravcová Effect of biologically active substances on laying traits of Japanese quails .................................................................... 138 Eliska Horecka, Cenek Horecky, Lenka Kovarikova, Ales Knoll, Ales Pavlík Single nucleotide polymorphism of PMCA genes in ISA Brown laying hens .............................................................. 139 Cyril Hrnčár, Denis Porubský, Ján Weis, Henrieta Arpášová, Veronika Pistová, Jozef Bujko The effect of supplementation of enzyme preparation on base of pepsin on performance and carcass characteristics of Japanese quail .......................................................................................................................................... 140 Inessa B. Izmailovich, Marcin W. Lis The adsorbents of mycotoxins in poultry .......................................................................................................................... 141 Michał Jóźwiak, Krzysztof Wyrostek, Edyta Świętoń, Monika Olszewska-Tomczyk, Krzysztof Śmietanka, Zenon Minta Trends in current research on avian influenza viruses .................................................................................................... 142 Dorota Katarzyńska, Małgorzata Grzesiak, Andrzej Sechman Selection of the best reference gene in real-time qPCR studies in ovarian tissue of hens treated with silver nanoparticles ......................................................................................................................................................................... 143 Joanna Klećkowska-Nawrot, Karolina Goździewska-Harłajczuk, Karolina Barszcz, Artur Kowalczyk, Ewa Łukaszewicz Microstructural features of the Harderian gland in Common pheasant (Phasianus colchicus talischensis) and hybrid of Italian Amber and Common pheasant ...................................................................................................... 144 Paweł A. Kołodziejski, Ewa Pruszyńska-Oszmałek, Maciej Sassek, Daniela Chałupka, Tomasz Szwaczkowski, Paweł Maćkowiak, Leszek Nogowski Symbiotic regulates incretin (GIP and GLP-1) expression and secretion in chicken duodenum .............................. 145 Ľubor Košťál, Mária Horváth, Katarína Pichová Optimising behavioural tests for studying cognitive bias in poultry ............................................................................. 146 Józefa Krawczyk Quality of native breed hen eggs stored under different conditions .............................................................................. 147 Józefa Krawczyk, Joanna Obrzut Carcass muscling and colour in poulards from Sussex (S-66) hens ............................................................................... 148 Józefa Krawczyk, Michał Puchała The effect of breed and free-range production system on quality of meat from hens of two native breeds ............. 149 10 Lucie Kupčíková, Martina Lichovníková, Alena Przywarová, Vojtěch Anderle Carcass quality of fattening laying-type cockerels ............................................................................................................ 151 Martina Lichovnikova, Sarka Volna The effect of day length on broiler breeders’ performance at suboptimal females’ weight ......................................... 152 Ewa Łukaszewicz, Martyna Lasoń Asymmetry of roosters’ primary and secondary sexual traits ........................................................................................ 153 Ewa Łukaszewicz, Martyna Lasoń, Murray Bakst, Krzysztof Andres, Joanna Rosenberger, Artur Kowalczyk Comparison of embryo development at the oviposition of three goose breeds ........................................................... 154 Mariana Máčajová, Monika Buríková, Ivan Čavarga, Pavel Výboh, Boris Bilčík The use of the quail chorioallantoic membrane as an experimental in vivo model ..................................................... 155 Renata Muchacka, Ewa Sosnówka-Czajka, Iwona Skomorucha Activity of antioxidant enzymes and degree of lipid peroxidation in eggs from Greenleg Partridge hens depending on the housing system ...................................................................................................................................... 156 Mangesh Nakade, Krisztián Balogh, Erika Zándoki, Zsolt Ancsin, Andrea Bócsai, Márta Erdélyi, Miklós Mézes Effects of herbal mixture and short-term aflatoxin exposure on performance, some blood lipid peroxide and glutathione redox parameters of laying hens ............................................................................................................. 157 Joanna Nowak, Katarzyna Połtowicz, Katarzyna Piórkowska Effect of age of broiler chickens on the expression level of genes coding for calpain (CAPN1, CAPN2, CAPN3) and calpastatin (CAST) in breast muscles ......................................................................................................................... 158 Laura Pardyak, Alicja Kamińska, Ewelina Górowska-Wójtowicz, Anna Hejmej, Mariola Słowińska, Małgorzata Kotula-Balak, Andrzej Ciereszko, Barbara Bilińska Expression of aromatase in the testis, epididymis and ductus deferens of the turkey (Meleagris gallopavo) with yellow semen syndrome .............................................................................................................................................. 159 Krzysztof Pawlak, Bartosz Bojarski, Marcin Lis, Weronika Kanik, Jerzy Niedziółka, Barbara Tombarkiewicz The influence of electromagnetic field emitted by mobile telephony on chick hematology parameters .................. 160 Csilla Pelyhe, Benjámin Kövesi, Erika Zándoki, Miklós Mézes, Krisztián Balogh, Balázs Kovács Combined effect of T-2 toxin and deoxynivalenol on regulation of the glutathione redox system in broiler chicken .................................................................................................................................................................................... 161 Katarína Pichová, Boris Bilčik, Ľubor Košťál Thermographic evaluation of feather cover damage in laying hens as a tool for welfare assessment ....................... 162 Angelika Podbielska, Anna Radko, Agnieszka Szumiec Applications of the CHD locus in sex identification in birds ......................................................................................... 163 Katarzyna Połtowicz, Joanna Nowak Effect of early feed restriction on liveability, rearing performance and meat quality of broiler chickens ................. 165 Andrzej Posyniak, Krzysztof Niemczuk Effectiveness and safety of the use of antibiotics in fowl ................................................................................................. 166 Ewa Pruszyńska-Oszmałek, Paweł A. Kołodziejski, Maciej Sassek, Daniela Chałupka, Tomasz Szwaczkowski, Paweł Maćkowiak, Leszek Nogowski In ovo synbiotic administration affects on metabolic and hormonal profile in broiler chickens ............................... 167 Anna Radko, Agnieszka Szumiec, Angelika Podbielska, Tomasz Borkowski Application of 10 microsatellites DNA markers for parentage control in pigeons ...................................................... 168 Joanna Rosenberger, Artur Kowalczyk, Ewa Łukaszewicz, Zenon Rzońca Nesting behavior of Western capercaillie (Tetrao urogallus) females in the breeding aviary ...................................... 169 Maciej Sassek, Ewa Pruszyńska-Oszmałek, Paweł A. Kołodziejski, Daniela Chałupka, Tomasz Szwaczkowski, Leszek Nogowski, Paweł Maćkowiak In ovo synbiotic administration modulates trypsin, lipase and amylase activity in broiler chicken ......................... 171 Iwona Skomorucha, Ewa Sosnówka-Czajka Effect of adding herb extracts to drinking water on broiler chicken productivity, corticosterone level and heterophil: lymphocyte ratio during heat stress ........................................................................................................ 172 11 Iwona Skomorucha, Ewa Sosnówka-Czajka Effect of housing systems on some blood parameters of broiler chickens during a period of high temperatures .......................................................................................................................................................................... 173 Joanna Socha, Andrzej Sechman, Maria Mika, Anna Hrabia Effect of growth hormone on estrogen receptor and egg shell-specific protein gene expression in the chicken shell gland during pause in laying induced by fasting ............................................................................ 174 Ewa Sosnówka-Czajka, Iwona Skomorucha Effect of adding herb extract to water on rectal and radiated temperature of laying hens during the summer heat .................................................................................................................................................................... 175 Ewa Sosnówka-Czajka, Iwona Skomorucha Effect of breed of laying hens on production results under organic conditions ........................................................... 176 Ewa Sosnówka-Czajka, Iwona Skomorucha, Eugeniusz Herbut Effect of housing system on some parameters of egg quality and egg-laying behaviour in laying hens ................... 177 Kinga Stuper-Szablewska, Tomasz Szablewski, Sebastian Nowaczewski, Renata Cegielska-Radziejewska The comparison of microbial transmission within intensive (commercial farm) and extensive housing systems on an annual basis ................................................................................................................................................... 178 Kinga Stuper-Szablewska, Tomasz Szablewski, Sebastian Nowaczewski, Anna Ostrowska-Kołodziejczak, Anna Matysiak, Renata Cegielska-Radziejewska The evaluation of the effect of seasons on the microbiological contamination of selected elements of the environment in terms of health safety of laying hens kept in extensive housing system .................................. 180 Barbara Tombarkiewicz, Karolina Trzeciak, Marcin Lis, Bartosz Bojarski, Krzysztof Pawlak, Weronika Kanik, Zbigniew Podgórni, Jerzy Niedziółka The effect of methionine in ovo administration on chick (Gallus gallus domesticus) embryo development ............ 182 Ján Weis, Cyril Hrnčár, Matej Rindoš, Slavomír Mindek Effect of probiotic application as a spray to egg shell on hatchability of broiler ducks ................................................ 183 Gabriela Wójcik, Magdalena Trela, Andrzej Sechman, Marcin W. Lis Changes in thyroid hormone concentrations in chick blood during hatching after in ovo α tocopherol administration ........................................................................................................................................................................ 184 Anna Wyrobisz, Ewa Sosnówka-Czajka, Jerzy Kowal, Paweł Nosal Comparison of parasite infection in laying hens kept in different housing systems .................................................... 185 Małgorzata Natonek-Wiśniewska, Piotr Krzyścin The method of identifying species of poultry on the basis of sequence differences in the s-rRNA ........................... 186 Urszula Kaczor, Marek Stanisz, Marta Bykowska, J. Składanowska, Dorota Wojtysiak, Andrzej Kaczor Fatty acid profile of intramuscular fat in supraspinatus, longissimus lumborum and semimembranosus muscles of farmed fallow deer .............................................................................................................................................. 187 Dorota Wojtysiak, Marek Stanisz, Agnieszka Ludwiczak, Urszula Kaczor Histochemical muscle fibre characteristics of fallow deer meat ...................................................................................... 188 Index of authors ............................................................................................................................................................................. 189 Plenary papers International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Resistin/adiponectin/FGF21/palmitic acid interplay in SH-SY5Y neural cells and DIO rodents: a novel mechanism contributing to insulin resistance Arieh Gertler The Institute of Biochemistry, Food Science and Nutrition, The Hebrew University of Jerusalem, Rehovot, Israel e-mail: arieh.gertler@mail.huji.ac.il Obesity and type2 diabetes (T2D), the two most serious diseases in the Western world share several features such as insulin resistance and energy homeostasis deregulation driven by changes of adipose tissue secreted hormones. Two such adipokines: adiponectin, an insulinsensitizing hormone, and in contrast resistin known to promote insulin resistance are thus potential links between obesity and T2D. In addition, Fibroblast Growth Factor 21 (FGF21), predominantly produced by the liver, has similar effects as adiponectin in regulating glucose and lipid metabolism and insulin sensitivity. High circulating levels of saturated free fatty acids such as palmitate promote inflammation and insulin resistance in peripheral tissues. In contrast, polyunsaturated fatty acids such docosahexaenoic acid (DHA) exhibit a protective action. The deleterious effects of palmitate have been attributed to its potential action through Toll-like receptor 4 (TLR4). However, the potential link between palmitate and resistin regarding TLR4 signaling at the neuronal level is unknown. In the present study, we investigated whether central resistin promotes insulin resistance through the impairment of adiponectin signaling and by promoting FGF21 resistance. Chronic ICV resistin infusion to rats down-regulated both hypothalamic and hepatic APPL1, a key protein in adiponectin signaling, associated with decreased Akt/APPL1 interaction and an increased Akt association with its endogenous inhibitor TRB3. Resistin treatment also reduced the expression of adiponectin receptors in hypothalamus, liver, muscle and adipose tissue. Furthermore central resistin acting through TLR4 impaired insulin sensitivity consequently to the downregulation of FGF21 and its receptor components in the hypothalamus and peripheral tissues promoting FGF21 resistance. We also showed that resistin effects are abolished in TLR4 knock-out mice and in SH-SY5y human neural cells expressing TLR4 siRNAs. In summary, our study reveals novel mechanism explaining the link between insulin resistance and central resistin/ TLR4 pathway that impairs adiponectin signaling and in parallel promotes FGF21 resistance. Palmitate increased TLR4 expression promoting resistin signaling through this receptor and its recruitment to membrane lipid rafts. These effects were prevented when neuronal cells were pretreated with DHA. Blocking resistin signaling by resistin antagonist developed by our group that leads to reduced obesity in mice fed high fat diet (HFD) and abolishing insulin resistance may thus serve as a novel tool for clinical intervention. Supported by ISF grant no. 476/15. 15 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Aktualne problemy dobrostanu drobiu Current problems of poultry welfare Roman Kołacz Katedra Higieny Środowiska i Dobrostanu Zwierząt, Uniwersytet Przyrodniczy we Wrocławiu e-mail: roman.kolacz@up.wroc.pl Polska należy do czołowych producentów mięsa drobiowego w Europie, zajmując pierwsze miejsce pod względem produkcji i eksportu mięsa z kurcząt brojlerów i trzecie miejsce pod względem produkcji i eksportu mięsa indyczego. Do 2010 r. odnotowywano w Polsce zwiększający się poziom produkcji jaj spożywczych, który w 2010 r. wynosił 10,1 mld sztuk. W 2012 r. udział kur niosek utrzymywanych w systemach klatkowych stanowił 87% ogółu pogłowia kur niosek (38,37 mln sztuk). Tylko 13% kur utrzymywanych było w alternatywnych systemach chowu (chów ściółkowy – 10,9%, chów wolnowybiegowy – 1,8%, chów ekologiczny – 0,2%), co na tle państw UE dało dla systemów pozaklatkowych jeden z niższych wskaźników. Powyższa struktura utrzymała się w 2013 r. W 2014 r. produkcja jaj w Polsce wyniosła 10,26 mld sztuk, tj. o 2,1% więcej niż rok wcześniej. Te dwa sektory produkcji drobiarskiej, tj. hodowla brojlerów kurzych i kur niosek jaj konsumpcyjnych, są w UE regulowane odpowiednimi przepisami dotyczącymi warunków dobrostanu tych ptaków, a intensywne metody chowu tych zwierząt wciąż jeszcze spotykają się z krytyczną opinią społeczną. W 2006 r. w badaniach „Eurobarometer 2006” dotyczących świadomości i wiedzy na temat dobrostanu zwierząt gospodarskich wśród obywateli UE z 25 państw 16 wykazano, że 68% respondentów twierdziło, iż dobrostan kur jest zły lub bardzo zły, a tylko 32% badanych – że jest dobry i bardzo dobry. Również 44% respondentów opowiedziało się za tym, aby w pierwszej kolejności poprawić dobrostan kur, a następnie brojlerów kurzych (42%). Podstawą krytyki dotychczasowych klatek bateryjnych było ograniczenie lub całkowite wyeliminowanie naturalnych zachowań kur, takich jak: ruch, grzebanie, poszukiwanie i pobieranie pokarmu, siadanie na grzędzie, gniazdowanie, rozpościeranie skrzydeł i trzepotanie skrzydłami, stroszenie piór, zażywanie kąpieli piaskowych lub ścieranie pazurków. Implikacje behawioralne a dobrostan kur i brojlerów kurzych Ograniczenie możliwości manifestowania wzorców zachowania się drobiu utrzymywanego w intensywnych systemach, w tym również w systemie bateryjnym, może prowadzić do cierpienia ptaków zarówno w sferze psychicznej, jak i fizycznej, co manifestuje się patologicznymi formami zachowania lub urazami i schorzeniami. I tak, u kur niemających możliwości grzebania i dziobania pokarmu z podłoża obserwuje się wzrost częstotliwości zachowań anormalnych, objawiających się wydziobywaniem piór u innych pta- ków i kanibalizmem. Pterofagię i kanibalizm znacznie częściej obserwuje się u kur w chowie bateryjnym niż w innych systemach. Wydziobywanie piór jest dla ptaków bolesne, towarzyszy mu często krwawienie, a odsłonięta skóra jest bardziej narażona na liczne urazy zewnętrzne. Nawyk ten znacznie częściej występuje w systemach, gdzie kury pozbawione są grzędy, oraz wtedy, gdy kurom podaje się paszę, która jest przez nie szybko zjadana. W naturalnych warunkach kury przeznaczają około 50% czasu na poszukiwanie i pobieranie pokarmu. Przyczyną pterofagii może być także pozbawienie kur kąpieli piaskowych. W utrzymaniu klatkowym uniemożliwiającym manifestację tego wzorca obserwuje się tzw. puste kąpiele piaskowe, co według Baxtera (1994) świadczy o dużej frustracji ptaków i ich obniżonym dobrostanie. Ważną przyczyną pterofagii u drobiu jest także niedobór białka w paszy, a głównie aminokwasów siarkowych i witaminy K. Zaburzenia behawioralne u kur prowadzą w intensywnych hodowlach do kanibalizmu manifestującego się wydziobywaniem fragmentów ciała (odsłonięta skóra, palce, stek, grzebień, dzwonki, a u indyków korale), którego przyczyny bywają bardzo zróżnicowane i są często następstwem pterofagii. Należy podkreślić, że częstość występowania kanibalizmu u kur w systemie bateryjnym jest niższa niż w systemach podłogowych. Jedną z dyskusyjnych metod zapobiegania kanibalizmowi jest obcinanie dziobów u kurcząt, powodujące zarówno ostry, jak i chroniczny ból u ptaków oraz objawy depresji. Zgodnie z obowiązującym ustawodawstwem unijnym, w celu uniknięcia pterofagii i kanibalizmu państwa członkowskie mogą zezwolić na przycinanie dziobów pod warunkiem, że wykonuje je odpowiednio wykwalifikowany personel, a wiek kurcząt przeznaczonych do użytkowania nieśnego lub kurcząt brojlerów nie przekracza 10 dni. Obecnie toczy się ożywiona dyskusja w krajach UE dotycząca wprowadzenia przez poszczególne kraje członkowskie dobrowolnego zakazu skracania dziobów. Taki zakaz obowiązuje już w Szwecji, Norwegii i Austrii, a 1 sierpnia 2016 r. będzie wprowadzony w Niemczech. Zakazu takiego nie wprowadziła Wielka Brytania, uzasadniając to tym, że skutki jego wprowadzenia byłyby gorsze dla dobrostanu ptaków. Zaleca się natomiast stosowanie w skracaniu dziobów metody mniej bolesnej: lasera podczerwieni zamiast rozżarzonego noża. James V. Craig pisał w „Domestic Animal Behavior” (1981), że „okaleczenie, stosowane w praktyce hodowlanej, takie jak np. przycięcie dzioba kurzego, jest opisywane z dużym ładunkiem emocji, jednak mimo że powoduje chwilowy ból osobnikom, daje wiele korzyści dla dobra całej grupy”. Innym objawem frustracji, a jednocześnie zachowaniem rekompensującym zachowanie normalne, jest tzw. pseudogniazdowanie lub inaczej zachowanie nieśne „puste”, obserwowane u kur utrzymywanych w klatkach bateryjnych. Pozbawienie kur możliwości gniazdowania wywołuje u nich duży niepokój, jest przyczyną ich cierpienia psychicznego i stanowi jeden z ważniejszych problemów behawioralnych kur. Implikacje zdrowotne a dobrostan kur i brojlerów kurzych Stan zdrowia kur zawsze świadczy o poziomie dobrostanu, ale i dobrostan ptaków wpływa na ich zdrowotność. Schorzenia będące następstwem obniżonego poziomu dobrostanu, a jednocześnie wynikające z systemu czy technologii utrzymania nazywane są technopatiami. Do najczęściej wyliczanych technopatii u drobiu należą schorzenia kończyn. Schorzenia te są opisywane u tych kur, które utrzymywano w systemach ograniczających im niezbędną powierzchnię do takich ćwiczeń ruchowych, jak spacer, bieganie, rozpościeranie skrzydeł. Wykazano, że wytrzymałość kości na złamania u kur z chowu bateryjnego jest o 41% niższa w porównaniu do kości kur z chowu tradycyjnego. Syndrom „klatkowego zmęczenia kur” będących w dobrej nieśności i kondycji, u których następuje nagłe przewrócenie się na grzbiet z towarzyszącym paraliżem, jest często opisywany literaturze naukowej. U kur tych obserwuje się kruchość i łamliwość kości długich, deformację mostka i żeber. Ograniczenie swobody ruchu widoczne szczególnie w klatkach bateryjnych jest główną przyczyną słabości kończyn, które manifestują się jako deformacje, osteoporoza, złamania czy dyschondroplazje oraz słabość mięśni. Konsekwencjami słabości kończyn może być zwiększona częstotliwość złamań, kulawizny, silny ból, niemożność pobierania pokarmu i wody, a przez to cierpienie z powodu głodu i pragnienia, wycieńczenie organizmu i śmierć. Do schorzeń kończyn powstających na tle warunków utrzymania należy zaliczyć uszkodzenia palców i stopy. Stwierdzono, że u kur w klatkach bateryjnych częściej występują: rogowacenia stopy, liczne jej pęknięcia, niekontrolowany przerost pazura i złamania lub zwichnięcia palców. Wyposażenie klatek w taśmy ścierne służące do ścierania pazurów oraz w gniazda i grzędy znacznie redukowało te uszkodzenia. Zdrowotność stóp i palców nie budzi natomiast żadnych zastrzeżeń w systemie ściółkowym, chyba że dochodzi do pogorszenia się jakości ściółki. Typowym schorzeniem występującym u brojlerów kurzych i indyczych jest zapalenie skóry podeszwy stopy (Foot Pad Dermatitis – FPD). Jest to stan zapalny przechodzący w zmiany nekrotyczne na powierzchni podeszwy stopy ptaka. Zmiany te powodują ból i obniżają 17 dobrostan. Ptaki dotknięte pododermatitis mogą kuleć, w przypadku bólu obserwuje się u nich niechęć do poruszania się oraz oszczędzanie chorej nogi. Przy zajęciu chorobą obu nóg ptaki leżą, zmniejsza się pobieranie pokarmu, następuje ich chudnięcie i osłabienie. Najważniejszym czynnikiem etiologicznym tego schorzenia jest jakość ściółki. Wilgotna ściółka powyżej 30%, powstająca w wyniku wadliwego systemu poidełek lub występujących biegunek, powoduje rozmiękczenie skóry podeszwy, co wpływa na jej większą podatność na uszkodzenia. Czynnikiem sprzyjającym podrażnieniom skóry jest także za niskie lub za wysokie pH ściółki. Na wysokie pH ma wpływ zwiększone stężenie amoniaku w powietrzu i ściółce, a na niskie – kwas moczowy pochodzący z odchodów lub podawanie w pokarmie większej ilości kwasów. Zarządzanie mikroklimatem w budynkach dla brojlerów w zakresie temperatury, wilgotności i wartości amoniaku w powietrzu jest ważnym elementem prewencji FPD. Niektórzy autorzy twierdzą, że zwiększona obsada brojlerów na m2 podłogi także może być czynnikiem ryzyka w etiologii pododermatitis, głównie ze względu na większe zawilgocenie ściółki i emisję amoniaku. Przedstawione zaburzenia behawioralne, liczne urazy i schorzenia powodujące cierpienie i śmierć kur w systemach bateryjnych nie były akceptowane przez opinię publiczną. W Europie organizowano częste protesty przeciwko systemowi klatkowego utrzymania kur. W wyniku tych protestów oraz licznych raportów naukowych wskazujących na cierpienie kur utrzymywanych w systemach bateryjnych, a także braku stosownych przepisów dotyczących warunków utrzymania kurcząt brojlerów Parlament Europejski dokonał rewizji dotychczasowych przepisów utrzymania kur niosek, wydając w 1999 r. nową dyrektywę oraz ustanawiając w 2007 r. dyrektywę dotyczącą utrzymania brojlerów kurzych. Prawne i ekonomiczne aspekty ochrony dobrostanu drobiu w UE i w Polsce Przepisy prawa europejskiego i polskiego w zakresie ochrony dobrostanu drobiu dotyczą: • warunków utrzymania, • warunków transportu, • warunków uboju i uśmiercania. W Unii Europejskiej prawo dotyczące utrzymania drobiu jest regulowane przez trzy dyrektywy: 1. Dyrektywa UE 1998/58 ustanawiająca minimalne standardy utrzymania zwierząt gospodarskich (tą dyrektywą objęty jest inny drób niż nioski i brojlery kurze). 18 2. Dyrektywa UE 1999/74 ustanawiająca podstawowe standardy utrzymania kur nieśnych, która zastąpiła obowiązującą do końca 2002 r. dyrektywę 88/166. 3. Dyrektywa Rady 2007/43/WE z dnia 28 czerwca 2007 r. w sprawie ustanowienia minimalnych zasad dotyczących ochrony kurcząt utrzymywanych z przeznaczeniem na produkcję mięsa. Dyrektywa Rady 1999/74/EC nie obowiązuje w stosunku do ferm użytkujących do 350 kur niosek i ferm utrzymujących zarodowe stada nieśne. Jednakże w stosunku do wspomnianych wyżej ferm obowiązują odpowiednie przepisy Dyrektywy 98/58/ EC, a w Polsce Rozporządzenie Ministra Rolnictwa i Rozwoju Wsi z dnia 28 czerwca 2010 r. w sprawie minimalnych warunków utrzymywania gatunków zwierząt gospodarskich innych niż te, dla których normy ochrony zostały określone w przepisach Unii Europejskiej (Dz. U. z 2010 r. Nr 116, poz. 778). Dyrektywa 99/74 określa warunki, jakim muszą odpowiadać klatki udoskonalone (często nazywane umeblowanymi), wprowadzane w miejsce dotychczas istniejących lub nowo budowanych ferm od 1 stycznia 2012 r. Kury w klatkach udoskonalonych muszą mieć zapewnione przynajmniej 750 cm2 powierzchni klatki na kurę, w tym 600 cm2 powierzchni dostępnej, niezabudowanej, a ogólna powierzchnia klatki nie może być mniejsza niż 2000 cm2. Ponadto klatki powinny być wyposażone w: • gniazdo, • ściółkę, umożliwiającą grzebanie i dziobanie, • grzędy (15 cm na kurę), • karmidła z dostępem bez ograniczeń (długość nie mniejsza niż 12 cm/kurę), • co najmniej dwa poidła kropelkowe lub kubeczkowe, dostępne dla każdej z kur znajdujących się w klatce lub 12 cm poidła rynienkowego przypadającego na 1 kurę, • urządzenia do skracania pazurów. Wymienione wyposażenie klatek powinno w pewnym stopniu zabezpieczać podstawowe potrzeby behawioralne kur. Wprowadzenie klatek udoskonalonych nie było automatyczną zamianą systemu klatek starego typu na nowe. W państwach UE spowodowało to duży spadek liczby kur utrzymywanych w systemach bateryjnych. W 25 państwach UE przed 2012 r. w klatkach bateryjnych starego typu (550 cm2/kurę) utrzymywanych było 79,6% kur, a obecnie utrzymywanych jest w systemach klatek wzbogaconych 56% kur. Na głębokiej ściółce utrzymywanych jest 26%, na wolnych wybiegach 14%, a w systemie ekologicznym 4%. W Polsce nadal obserwuje się wysoki wskaźnik utrzymania kur w systemie bateryjnym (88%), na głębokiej ściółce 2%, na wolnym wybiegu 10%, a w systemie ekologicznym zaledwie 0,002% kur. Obowiązek wdrożenia od 1 stycznia 2012 r. Dyrektywy 99/74 był znacznym obciążeniem finansowym dla hodowców. Modernizacja jednego stanowiska dla kury kosztowała 10–15 euro. Wzrósł także koszt produkcji, powodując nierówną konkurencję cenową na rynku europejskim, np. w zakresie proszku jajecznego importowanego z krajów trzecich, gdzie standardy utrzymania kur w bateriach są dalekie od europejskich. Minimalna powierzchnia klatki w USA i Kanadzie wynosi 430 cm2/kurę, a w Brazylii, Indiach i na Ukrainie zaledwie od 350 do 400 cm2 na kurę, czyli prawie dwa razy mniej niż w UE. Kolejne negocjacje WTO z Komisją Europejską zmierzające do zniesienia barier celnych ze względu na dobrostan zwierząt oraz 13. runda negocjacji w sprawie porozumienia handlowego pomiędzy UE a USA o wolnym handlu (TTIP) mogą być niekorzystne dla rolnictwa europejskiego, a szczególnie sektora drobiarskiego. Sytuacja ta dotyczy także mięsa drobiowego. Poubojowa ocena dobrostanu kurcząt brojlerów Przepisy Dyrektywy Rady 2007/43/WE w sprawie ustanowienia minimalnych zasad dotyczących ochrony kurcząt utrzymywanych z przeznaczeniem na produkcję mięsa weszły w życie w Polsce 30 czerwca 2010 r. na mocy Rozporządzenia Ministra Rolnictwa i Rozwoju Wsi (Dz. U. z 2010 r. Nr 56, poz. 344). Najważniejszym wymogiem tej dyrektywy jest zapis, aby maksymalna obsada kurcząt brojlerów w kurniku w żadnym momencie nie przekraczała 33 kg/m2. W formie odstępstwa od powyższej zasady zezwala się na zwiększenie obsady do 39 kg/m2 lub 42 kg/m2 pod warunkiem spełnienia dodatkowych wymogów określonych w dyrektywie. Zwiększona obsada do 42 kg/m2 wymaga zgody Powiatowego Lekarza Weterynarii, która może być wydana tylko wtedy, gdy wszystkie kontrole dobrostanu w kurniku są pozytywne, gdy skumulowany wskaźnik śmiertelność kurcząt nie przekracza wymaganej dyrektywą wartości oraz gdy zdrowotność kurcząt w badaniach poubojowych oceniana przez urzędowego lekarza weterynarii w rzeźni, na podstawie m.in. zmian na podeszwie łapek powstałych w wyniku pododermatitis, nie budzi jego zastrzeżeń. Ocenę tych zmian przeprowadza się na podstawie badania jednej łapki od 100 ptaków z jednego stada utrzymywanego w tym samym kurniku. Skala oceny to: • 0 – brak zmian, • 1 – powierzchowne zmiany, przebarwienia, • 2 – głębokie zmiany ze strupem i owrzodzeniem. Po dokonaniu oceny zmian według powyższego kryterium sumuje się liczbę punktów otrzymanych z poniższych iloczynów dla całej przebadanej partii 100 łapek: • liczba łapek ocenianych na 0 jest mnożona przez 0, • liczba łapek ocenianych na 1 jest mnożona przez 0,5, • liczba łapek ocenianych na 2 jest mnożona przez 2. Końcowa ocena partii łapek zależy od ogólnej liczby punktów. Jeżeli ogólna liczba punktów: • < 40 – bez uwag, • 41–80 – niski poziom nieprawidłowości, • 80 – wysoki poziom nieprawidłowości. Końcową ocenę poziomu dobrostanu odstawionych do ubojni kurcząt brojlerów przyjmujemy jako: • dobry poziom dobrostanu, gdy: skumulowany wskaźnik śmiertelności jest zgodny z przepisami (1% + 0,06% pomnożony przez wiek stada w dniu uboju podany w dniach) i ocena zmian na łapkach wynosi mniej niż 40 pkt, • średni poziom dobrostanu, gdy: skumulowany wskaźnik śmiertelności dziennej jest mniejszy od 5%, przy braku uzasadnienia takiego poziomu śmiertelności, lub/i ocena zmian na łapkach wynosi 41–80 pkt, • niski poziom dobrostanu, gdy: skumulowany wskaźnik śmiertelności dziennej jest wyższy niż 5%, przy braku uzasadnienia takiego poziomu śmiertelności, lub/i ocena zmian na łapkach jest większa niż 80 pkt. W przypadku stwierdzenia średniego lub niskiego poziomu dobrostanu urzędowy lekarz weterynarii informuje powiatowego lekarza weterynarii nadzorującego rzeźnię o wynikach badania i przesyła komplet dokumentacji dla badanej partii. Powiatowy lekarz weterynarii nadzorujący rzeźnię informuje powiatowego lekarza weterynarii nadzorującego gospodarstwo pochodzenia kurcząt o wynikach badania, przesyłając otrzymaną dokumentację. Po otrzymaniu powyższej informacji powiatowy lekarz weterynarii właściwy dla gospodarstwa pochodzenia kurcząt: • w przypadku stwierdzenia po raz pierwszy średniego poziomu dobrostanu w stadzie pochodzącym z danego gospodarstwa rozważa konieczność przeprowadzenia kontroli dobrostanu kolejnego stada utrzymywanego w danym gospodarstwie, biorąc pod uwagę dotychczasowe wyniki kontroli, wysokość zagęszczenia brojlerów oraz wielkość stada, • w przypadku stwierdzenia średniego poziomu dobrostanu po raz drugi dla przesyłki pochodzącej z innego stada brojlerów z tego samego gospodarstwa lub w przypadku stwierdzenia niskiego po- 19 ziomu dobrostanu przeprowadza kontrolę dobrostanu kolejnego stada utrzymywanego w danym gospodarstwie. Dobrostan drobiu a bezpieczeństwo żywności Utrzymanie wysokiego standardu dobrostanu zarówno kur, jak i kurcząt brojlerów ma szczególne znaczenie ze względu na bezpieczeństwo żywności. Chociaż na pozór związek pomiędzy poziomem dobrostanu drobiu a bezpieczeństwem żywności wydaje się odległy, to zagadnienia dobrostanu zwierząt zarówno w prawodawstwie UE, jak i w świadomości oraz oczekiwaniach konsumentów wysuwają się obecnie na pierwszy plan. Należy stwierdzić, że obniżonemu poziomowi dobrostanu zwierząt towarzyszy zawsze obniżona ich zdrowotność, i odwrotnie: chore zwierzę charakteryzuje się zawsze obniżonym dobrostanem. Jeżeli zatem przyjmiemy, że bezpieczna żywność może pochodzić tylko od zdrowych zwierząt, to zależność między dobrostanem zwierząt a bezpieczeństwem żywności nie jest ani pozorna, ani odległa. Obniżony poziom dobrostanu drobiu sprzyja zawsze powstawaniu chorób zakaźnych, warunkowo zakaźnych lub technopatii. Wysoki poziom dobrostanu to jednocześnie gwarancja dobrego stanu zdrowia zwierzęcia, a także eliminacja antybiotyków czy innych 20 lekarstw stanowiących potencjalne zagrożenie dla bezpiecznej żywności. Obniżenie poziomu dobrostanu drobiu ze względu na nieodpowiednie warunki utrzymania czy transportu może w wyniku immunosupresji doprowadzić do zakażenia całego organizmu drobnoustrojami żyjącymi w przewodzie pokarmowym, stwarzając kolejny element ryzyka dla bezpiecznej żywności. Generalnie stwierdza się, że produkowana obecnie żywność nie jest całkowicie bezpieczna. Ale jednocześnie nigdy wcześniej nie była bezpieczniejsza niż teraz, chociaż jej bezpieczeństwo nie było kwestionowane bardziej niż obecnie. Ta nieufność konsumentów nie jest bezpodstawna. Liczne doniesienia epidemiologiczne z ostatnich lat w Europie i na świecie wskazują, że wzrasta liczba czynników ryzyka zdrowia zwierząt, a przez to i liczba ognisk chorobowych. Równocześnie wprowadzane ostatnio daleko idące zmiany w systemie produkcji drobiarskiej nie dają całkowitej gwarancji, że mięso i jaja będą pozbawione zarazków oraz pozostałości po antybiotykach i substancjach obcych obecnych często w skażonym środowisku. Dlatego ochrona dobrostanu drobiu odgrywa podstawową rolę w produkcji bezpiecznej żywności, a przez to i w ochronie zdrowia publicznego. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Chicken ovary – a place of dioxin and polychlorinated biphenyl detoxification Andrzej Sechman, Piotr A. Antos, Dorota Katarzyńska, Anna Hrabia Department of Animal Physiology and Endocrinology, University of Agriculture in Krakow, Poland e-mail: rzsechma@cyf-kr.edu.pl Dioxins and polychlorinated biphenyls (PCBs) are chloroorganic compounds which belong to persistent environmental pollutants and endocrine disruptors. In the domestic hen (Gallus domesticus) they exhibit a diverse spectrum of unfavourable effects in the embryonic [3] and adult ovary [1, 2, 5]. In a target cell, dioxin and PCBs bind to the aromatic hydrocarbon receptor (AHR), translocate to the nucleus and induce a cytochrome P-4501A monooxygenase system which is involved in the first step of xenobiotic metabolism [4, 6]. To clarify whether chicken ovarian follicles may participate in dioxin and PCB detoxification, the effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 3,3’,4,4’,5-pentachlorobiphenyl (PCB 126; co-planar PCB congener) and 2,2’, 4,4’,5,5’-hexachlorobiphenyl (PCB 153; non-coplanar PCB congener) on mRNA expression of AHR1, CYP1A5 and CYP1A5 genes as well as their activity (measured by EROD and MROD activities) in chicken ovarian follicles were investigated. The white (WF) and yellowish (YF) nonhierarchical follicles and fragments of the theca (TL) and granulosa (GL) layers of the 3 largest yellow preovulatory follicles (F3–F1) were isolated from the chicken ovary and incubated in a medium supplemented with 1, 10 or 100 nM TCDD (n = 6), PCB 126 (n = 6) or 1, 10 or 100 µM PCB 153 (n = 6). In the collected tissues, expression of AHR1, CYP1A5 and CYP1A5 was measured by real-time qPCR while EROD and MROD activities by fluorescence methods. Results were statistically evaluated by ANOVA followed by Tukey test at P < 0.05. The experiment confirmed that chicken ovarian follicles express AHR1, CYP1A5 and CYP1A5. In WF and YF a stimulatory effect of all three xenobiotics on AHR1 mRNA levels was found. On the other hand, in the TL and GL of F3-F1 follicles a down-regulation of AHR1 following TCDD exposition was observed. PCB 153 stimulated AHR1 expression in the WF and the GL of F1 follicle. A strong stimulatory effect of TCDD and PCB 126 on CYP1A5 in the YF, and the TL and GL of F3–F1 follicles was noticed; the effects of these two chemicals on CYP1A5 expression were relatively lower. PCB 153 was able to stimulate these genes expression but only in the GL of the F1 follicle. All investigated chemicals in dose-dependent manner increased EROD and MROD activities in the chicken ovary in the following order: TCDD > PCB 126 > PCB 153. In summary, data of this experiment revealed that the chicken ovarian follicles express AHR1, CYP1A5 and CYP1A5 genes, and dioxin and PCBs modulate their transcription and stimulate EROD and MROD activity. It is suggested that chicken ovarian follicles are a place of dioxin and PCB detoxification. Supported by grants: N N303 561 339 and DS-3243/ KFiEZ. 21 References [1] Antos P.A., Błachuta M., Hrabia A., Grzegorzewska A.K., Sechman A. 2015. Toxicol. Lett., 237, 100–111. [2] Sechman A., Antos P., Katarzyńska D., Grzegorzewska A., Wojtysiak D., Hrabia A. 2014. Toxicol. Lett., 225, 264–274. [3] Sechman A., Hrabia A., Lis M.W., Niedziółka J. 2011. Toxicol. Lett., 205, 190–195. 22 [4] Sorg O. 2014. Toxicol. Lett., 230, 225–233. [5] Wójcik D., Antos P.A., Katarzyńska D., Hrabia A., Sechman A. 2015. Toxicol. Lett., 239, 73–80. [6] Yasui T., Kim E.Y., Iwata H., Franks D.G., Karchner S.I., Hahn M.E., Tanabe S. 2007. Toxicol. Sci., 99,101–117. Session 1 Animal welfare and product quality International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Roles, tasks and responsibilities of the Designated Veterinarian under Directive 2010/63/EU Piotr SJ Sowinski European Society Laboratory Animal Veterinarians (ESLAV) P Sowinski, DVM, LAVA Department of Bioengineering, Royal School of Mines Building, Imperial College London, South Kensington Campus, United Kingdom e-mail: p.sowinski@imperial.ac.uk Work and use of animals has been recently regulated on a European level by a new legislative act called Directive 2010/63/EU. The Directive requires that a range of veterinary services should be provided at all level and at all time. Directive 2010/63/EU provides for veterinarians a legal frame work to continue fulfilling this responsibility. Laboratory Animal Veterinarians are involved in a broad range of activities related to the functions of Ethical Committees, facility management, health monitoring, research projects, compliance with legislation and the training of personal and researchers. Considering the variety of sizes of establishments, the range of species, and the type of experiments for which animals may be used, it is in the view of European professional organisations, i.e.: the European Society of Laboratory Animal Veterinarians and the European College of Laboratory Animal Medicine that the veterinary care programme and the training of the designated veterinarian should be tailored to the specific needs of the environment in which they work. This presentation will be in the form of a discussion regarding whether the veterinarians are able to fulfil the obligations of the Directive as well as the expectations of animal users without compromising animal welfare. 25 Rola, zadania i odpowiedzialność wyznaczonego lekarza weterynarii w świetle Dyrektywy 201063/EU Piotr SJ Sowinski European Society Laboratory Animal Veterinarians (ESLAV) P Sowinski, DVM, LAVA Department of Bioengineering, Royal School of Mines Building, Imperial College London, South Kensington Campus, Wielka Brytania e-mail: p.sowinski@imperial.ac.uk Nowa europejska Dyrektywa 2010/63/EU wymaga od organizacji pracujących ze zwierzętami laboratoryjnymi udziału służby weterynaryjnej, regulując zadania i odpowiedzialność w zakresie doradczym. Z danych dostępnych z europejskich organizacji zawodowych wynika, że obecnie ponad 1000 lekarzy jest zatrudnionych w sektorze medycyny zwierząt laboratoryjnych. Zasiadają oni w komisjach etycznych, są członkami kierownictwa zwierzętarni, zajmują się monitoringiem zdrowia zwierząt, biorą udział w opracowaniu projektów naukowych, kontrolują przestrzeganie zapisów legislacyjnych, są członkami komisji bezpieczeństwa i higieny pracy, a także zajmują się szkoleniem zawodowym. Powinnością zawodową, etyczną i prawną weterynarza jest ochrona zdrowia zwierzęcia i jego dobrostanu. Istnieje zatem 26 potrzeba, aby lekarz weterynarii był włączony w opracowanie i wdrażanie programów odpowiedniej opieki nad zwierzętami. Biorąc pod uwagę wielkość placówek naukowych, złożoność prowadzonych tam prac badawczych, a także rozpiętość używanych gatunków zwierząt, rodzi się pytanie, czy lekarz wykształcony w zawodzie weterynaryjnym może sprostać tym wymaganiom, czy też program specjalistycznego kształcenia lekarza weterynarii powinien być dopasowany do potrzeb środowiska, w jakim przyjdzie mu pracować. Prezentacja jest formą dyskusji na temat funkcji i odpowiedzialności lekarza weterynarii w placówkach naukowo-badawczych, jego roli w ocenie projektów badawczych, a także w szkoleniu personelu i doradzaniu pracownikom naukowym. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Fatty acid profile of lamb meat from highly productive breeds of sheep reared under organic conditions Paweł Paraponiak Department of Animal Genetic Resources Conservation, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: pawel.paraponiak@izoo.krakow.pl The aim of the study was to compare the fatty acid content of meat from highly productive breeds of Suffolk and Pomeranian sheep, reared in an organic production system. The profile of fatty acids, including conjugated dienes of linoleic acid (CLA), was analysed in the samples of longissimus dorsi muscle collected from Suffolk (S) and Pomeranian sheep (P) ram-lambs (14 animals per group; experimental slaughter at 120 days of age). The lambs were pastured from late April/early May and forage formed the basis of their diet. The animals were also supplemented with ground cereals (approx. 0.40 kg/day/animal). The results showed similar values for both groups as well as a higher proportion of unsaturated fatty acids (in the region of 60 g/100 g of all analysed fatty acids) compared to saturated fatty acids (more than 40 g/100 g of all analysed fatty acids). The most abundant saturated fatty acids (SFA) were palmitic acid (C6, 23–24%) and stearic acid (c18, 12–13%). Oleic acid (c18-1) showed the highest concentration among both monounsaturated fatty acids (MUFA, 95%) and unsaturated fatty acids (UFA, 67–72%). The meat of P lambs had a significantly higher proportion of PUFA, PUFA-6 (P ≤ 0.05) and PUFA-3 (P ≤ 0.01). PUFA 6/3 was 7.6 in group S and 6.8 in group P. CLA content of S and P meat was rather low at 1.084 and 1.147 g/100 g of all analysed fatty acids, respectively. The average configuration of the health-promoting parameters of meat from experimental ram-lambs was probably the outcome of the considerable proportion of concentrates in the ration and the relatively short period of feeding pasture forage (higher weight gains and, as a consequence, the earlier achievement of required preslaughter weight reduced the pasture rearing period). 27 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The influence of mycotoxin’s adsorbent of new generation “Funginorm” on the productivity of fattening pigs Victoria I. Borodulina, N.A. Sadomov The Belarusian State Agricultural Academy, Gorki, Belarus e-mail: helenamikulich1971@gmail.com The infection of grain and animal’s feed with mycotoxins is currently a major problem of modern pig breeding. This problem causes considerable economic damage to the industry. Mycotoxins are products of the metabolism of fungi that infect corn and other feed crops. The pollution of grain with mycotoxins is possible at all stages of its production. The prevalence of mycotoxins in nature is considered as a global problem. However, in some regions of the world, some mycotoxins occur more often than others. In conditions of man-made stresses an urgent task is to find ways and means to increase the protective forces of an organism of animals, contributing to increase the efficiency. To carry out scientific experiment on the principle of analogues were selected 80 pigs. Fattening pigs were divided into 4 groups of 20 animals each. Average live weight of pigs was 55.9– 57.8 kg. In the control group used only the basic diet for feeding fattening pigs. In the 1st test group in the basic ration was added 0.5 kg/ton of “Funginorm” is an adsorbent of a new generation. In the 2nd test group was added – 1.0 kg/ton of adsorbent and in the third experimental group was added – 1.5 kg/ton of adsor- 28 bent. As the main diet for guinea pigs used the SC-26 feed. “Funginorm” is adsorbent of a new generation for poultry and pigs, used to suppress the development of molds and neutralization of mycotoxins in feed. As controlled indexes for fattening pigs of all experimental groups used their body weight, average daily gains and feed conversion. Results of researches for 1st month indicate that the most pronounced advantage in growth rate of pigs was detected in the second and the third experimental groups. The average daily weight gain of pigs second and third experimental groups through the study month was 580 g and 577 g, which is 11.5% and 11.0% was significantly higher than in the control group. The average daily weight gain was 550 g, which is higher than in the first test group than in control at 30 g. For the second month of the test to maintain the achieved advantage intensity body weight gain guinea pigs for fattening. The second experimental group of fattening pigs had average daily gain – 661 g, 12.2% was significantly higher than in the control group. The third test group average daily weight gain was 642 g, which was significantly higher than in the control group of 54 g, respectively. Preservation of fattening pigs all experimental groups was 100%. In general, for the entire period of the experience the average daily weight gain in the second test group was 622 g, which is 11.9% significantly higher than in control group. In the third test group average grain was 611 g, an increase of 9.9% was significantly higher compared with the control group. The inclusion of adsorbent in fattening pigs diet at doses of 0.5–1.5 g/kg of feed conversion and reduced feed conversion ratio increased feed of 3.5–12.4% compared with the control group. Throughout the experiment, the growth of the most energy intensive fattening pigs had two and three experimental groups that received a new generation of adsorbent “Funginorm” in an amount of 1.0 g/kg and 1.5 g/kg feed. They were superior to their peers in the control group to 11.9% and 9.9% respectively. The feeding of fattening pigs, in order to prevent, reduce the action of mycotoxins in feeds and enhance the productive performance. We recommend the use of a new generation of adsorbent «Funginorm» in doses of 1.0–1.5 g/kg of feed. 29 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Mycotoxins in dairy cows diet in south-west Bohemia Veronika Čoudková, J. Starý, Miroslav Maršálek Department of Animal Husbandry Sciences, Faculty of Agriculture, University of South Bohemia in České Budějovice, Czech Republic e-mail: nika.13@seznam.cz Mycotoxins may caused problems with health and production especially in the transition period of dairy cows. The aim of the study was to determine the amount of the risk of contamination of feed for dairy cows by three most surveyed mycotoxins, also find and indicate the type of feed with the potentially highest risk. Three major groups of mycotoxins were studied, Zearalenone (ZEA) for its hyperoestrogenic effects, T- toxin and Deoxynivalenol (DON) for immunosuppressive effects, epitheliotoxicity and hepatotoxicity. A total of 226 feed samples were provided by farmers from South-west region of the Czech Republic. The material was divided into four groups: grain mixes (n = 49), corn silage (n = 65), grass silage (n = 86) and hay (n = 26). Mycotoxin analyses were performed by ELISA method. All of samples were positive for the monitored mycotoxins. The highest levels of mycotoxins were detected in DON: 1529 µg/ kg feed in grain mixes, 1329 µg/kg feed in corn silage; T-2 was 968 µg/kg feed in grain mixes, 932 µg/kg feed in corn silage and 924 µg/kg feed in grass silage. The highest rate of overlimit concentrations of all studied mycotoxins was detected in corn silage, where 46% of the samples contained overlimit amount of DON 30 (limit 200 µg/kg feed), 35% samples of T-2 (limit 200 µg/kg feed) and 22% of ZEA (limit 100 µg/kg feed). The observed levels of mycotoxins correspond to occurence of appropriate mycotoxins in other areas of Europe. Based on the results the estimation of theoretical intake of mycotoxins in daily ration can be done. With model ration (14 kg grass silage, 22 kg corn silage and 5 kg grain mixes) dairy cows can consume up to 48.027 mg of DON per day, what demonstrate risk of total daily amount of consumed mycotoxin, which may caused decrease in production and feed intake The maximum acceptable levels of mycotoxins contamination must be strict regarding at the material used in animal nutrition, especially in roughage. Regular measurements of mycotoxins levels and early elimination of contaminated feed leads to reduction of the negative impacts on the health and production of dairy cows. This work would not have been possible without the financial support of Grant Agency of University of South Bohemia GAJU-019/2016/Z „Výživa a technika chovu jako cesta k pohodě zvířat a kvalitě produktů”. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Selected reproductive indicators for mares of the Hucul Pony breed Bogusława Długosz1, Romana Augustyn1, Magdalena Pieszka1, Jarosław Łuszczyński1, Tomasz Próchniak2, Barbara Jaklińska3, Karina Świderska1, Zenon Podstawski1, Monika Stefaniuk-Szmukier1, Weronika Pisarczyk1 1 Department of Horse Breeding, Institute of Animal Sciences, University of Agriculture in Krakow, Poland 2 Department of Biological Bases of Animal Production, University of Life Sciences in Lublin, Poland 3 Agriculture Farm „Kietrz” Sp. z o.o., Hucul Horse Stud „Gładyszów” e-mail: b.dlugosz@ur.krakow.pl Mares females are seasonally polyestral and showing reproductive activity during the long day, and loss of activity in the winter. Knowledge of the course of the seasonal cycles of sex mare is very important from the point of view of breeding, as appropriate determination of the time of ovulation increases the chances of fertilization of mares in the first estrus and foals, prior and following weaning from their mothers at the beginning of the grazing season. Foals with access for a long time for a fresh, green feed are stronger and more resistant and have better overall condition. The main objective of this thesis is to calculate each of the reproductive indicators for 238 mares of the Hucul Pony breed in years the 2000–2009 in the Hucul Stud “Gładyszów”. The pregnancy rate for the last 10 years is 87.3%, that is 10.9% more than in previous years. The fertility indicator reached 81.2%, and that value is also 8.3% larger than in previous years, but unfortunately the miscarriage indicator increased as well, reaching the average value of 4.6% – that is 0.6% more in comparison to calculations made in years the 1990– 2000. In 2001 the number of ponies separated from their mothers was the biggest – it reached 97.2%. The worse year in context of the results was 2009, because the number of unfertilized mares was the largest and the number of resorptions and registered dead ponies younger than 6 months was greater than in previous years as well. 31 Wybrane wskaźniki rozrodowe klaczy huculskich Bogusława Długosz1, Romana Augustyn1, Magdalena Pieszka1, Jarosław Łuszczyński1, Tomasz Próchniak2, Barbara Jaklińska3, Karina Świderska, Zenon Podstawski1, Monika Stefaniuk-Szmukier1, Weronika Pisarczyk1 Zakład Hodowli Koni, Instytut Nauk o Zwierzętach, Uniwersytet Rolniczy w Krakowie Katedra Biologicznych Podstaw Produkcji Zwierzęcej, Uniwersytet Przyrodniczy w Lublinie 3 Kombinat Rolny „Kietrz” Sp. z o.o., Stadnina Koni Huculskich „Gładyszów” 1 2 e-mail: Klacze to samice sezonowo poliestralne, wykazujące aktywność rozrodczą w czasie długiego dnia, a następnie zanik tej aktywności w zimie. Znajomość przebiegu sezonowości cykli płciowych klaczy jest bardzo istotna z punktu widzenia hodowlanego, ponieważ właściwe określenie terminu owulacji zwiększa szanse na zapłodnienie klaczy już w pierwszych rujach i wcześniejsze uzyskanie źrebiąt, a w następstwie odsadzenie ich od matek na początku trwania sezonu pastwiskowego. Źrebaki mające dostęp przez dłuższy czas do świeżej, zielonej paszy są silniejsze i bardziej odporne oraz mają lepszą ogólną kondycję. Celem pracy było obliczenie poszczególnych wskaźników rozrodu u 238 klaczy rasy huculskiej w Stadninie Koni Huculskich „Gładyszów”. Analizie zostały poddane dane dotyczące rozrodu klaczy w latach 2000–2009 pochodzące z dokumentacji hodowlanej stadniny takiej jak: karty klaczy, wykazy wyźrebień i stanówki. Określono następujące wskaźniki: źrebność, płodność, odsadzenia, jałowość, poronienia. 32 W badanym okresie średni wskaźnik źrebności wyniósł 87,3% i w stosunku do lat poprzednich uległ znacznemu podwyższeniu. Wskaźnik płodności, którego średnia w badanym okresie wyniosła 81,2%, zwiększył się w stosunku do uprzedniego dziesięciolecia, co świadczy o odpowiednim określeniu terminu krycia i dobrym doborze ogiera kryjącego. Średni wskaźnik odsadzeń dla badanych 10 lat wyniósł 87,8%. Niekorzystnie wzrósł wskaźnik poronień, kształtując się na średnim poziomie 4,6%, wyższym o 0,6% w stosunku do wskaźnika obliczonego w latach 1990–2000 (Osinka 2001). Najgorszym rokiem pod względem badanych wyników był rok 2009, kiedy to najwięcej klaczy jałowiło. W stosunku do lat ubiegłych również najwięcej było resorbcji i strat wśród źrebiąt do 6. miesiąca życia. Należy zwrócić większą uwagę na przyczynę wzrostu poronień, martwych urodzeń i upadków źrebiąt. Średnią wartość wskaźników rozrodu u klaczy z SKH „Gładyszów” można uznać za dobrą. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Bioaccumulation and elimination of cadmium in tissues of Prussian carp (Carassius gibelio B.) under controlled conditions Ewa Drąg-Kozak, Ewa Łuszczek-Trojnar, Magdalena Socha, Grzegorz Gosiewski, Włodzimierz Popek Department of Ichthyobiology and Fisheries, University of Agriculture in Krakow, Poland e-mail: e.drag-kozak@ur.krakow.pl Cadmium (Cd) is widely distributed in the aquatic environment and can cause severe damage to aquatic vertebrates. The aim of the work was to determine the accumulation level of Cd in scales and bone of Prussian carp (Carassius gibelio B.) during 7 weeks period of the exposure to three doses (0.04, 0.4 and 4 mg/L) of that metal in the water, and the effect of 3 weeks elimination period (after the previous 4 weeks exposure to that metal). Fish were divided into 4 groups: one control group (in water without the cadmium) and 3 experimental groups with addition of cadmium (0.04, 0.4 and 4 mg/L). After the period of 4 weeks of exposure all experimental groups were divided into two groups. One of them was still in Cd-polluted water (the same concentration), and the second one was transferred into un-polluted water for 3 weeks. After the every week of the experiment a random sample of 8 fish of each group were taken to measure Cd level in tissues. All samples were analyzed by AAS method using an atomic absorption spectrometer ATI UNICAM 929. Cadmium has been accumulated in the scales and bone since the beginning of the exposure because already after one week, the level of Cd significantly increased in all groups. The highest levels of cadmium were observed in the scales after one week of exposure (1.37 mg/kg) while in the bone after 5 weeks (1.32 mg/kg). The decrease of cadmium concentration in the scales and bone was observed after two and three weeks of exposure. A clear depuration of scales and bone was already observed at one week after termination of exposure. The low elimination was observed for the bone. Until the end of the experiment highly significant differences were observed in the scales and bone in the group which was exposed to the highest level of cadmium. Scales collected from live fish can be used for monitoring contamination of fish. This research was financed by the Ministry of Science and Higher Education of the Republic of Poland. 33 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The analysis of welfare Visayan warty pig (Sus cebifrons) kept in selected zoos Jan Dybała, Aleksandra Milewska, Joanna Wiśniewska, Aleksandra Cebulska, Wojciech Kapelański Department of Swine and Horses Breeding, University of Science and Technology in Bydgoszcz, Poland e-mail: dybala@utp.edu.pl Among the many tasks of zoos, the most important are: keeping rare animals, even of endangered species and their protection, as well. The reproduction of these animals determines maintenance of species and even their introduction to natural environment. The aim of the study was to assess the welfare of Visayan warty pig in zoos in Łódź and in Poznań and compare them with the applicable law in Poland. In both zoos 4 Visayan warty pigs were kept. The area of internal enclosures in Łódź and in Poznań was 12 m2. The area of external enclosures was respectively 280 m2 and 800 m2. The feed rations in both zoos were very similar: carrots, apples, beetroot, potatoes, seasonal fruits (eg. strawberries, plums), pears, oat flakes, boiled rice and corn, minced meat or one day old chicks, boiled eggs (2 pieces), cottage cheese were in their composition. 34 Visayan warty pig is quite rare species because lives only on two Philippine islands. The natural habitat of occurrence of this animal are dense forests. This animal is omnivorous, eating fruits, leaves, and animals, generally invertebrate. According to the Regulation of the Polish Minister of the Environment (Dz. U. 2005, Nr 5, poz. 32) the area of internal enclosure should be of 6 m2 for a couple + 10% for each additional animal. In both zoos the ground of enclosure is soft, allowing digging. There are also tree trunks, snag, that allow animals for hiding. The feed ration has very similar composition in both zoos, corresponding to their natural food preferences. Respectively in zoo in Łódź and in Poznań the animals have permanent access to water. These data shows that the welfare of Visayan warty pig in both analyzed zoos is fully preserved. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The influence of frequency of enrichments replacement on behavior of laboratory guinea pigs Kinga Fic, Aleksandra Woszczenko, Agnieszka Krzystanek, Natalia Sowińska Center of Experimental and Innovative Medicine, University Centre of Veterinary Medicine UJ-UR, Krakow, Poland e-mail: kinga.fic07@gamil.com Ensuring an appropriate level of animal welfare is an important element that has a direct impact on the animals and thereby affects the outcome of animal experiments. Enriching the environment of the animals through various items is a compulsory part of their maintenance. Adequate organization of the rotation of the enriching elements provides behavioral stimulation of animals and thus maintain a high level of animal welfare. The aim of the study was to determine how the frequency of replacement of three types of enrichments: berry discs, rolls of cardboard and wooden blocks contributed on the behavior of the guinea pigs. For 30 days, two groups of animals of 10 individual each, were recorded. In the first group items were exchanged with a higher frequency (four times a week), in the second with lower frequency (twice a week). In total, 33 hours of video materials were collected. Recordings were carried out in an irregular manner at different times of the day. The occurrence of particu- lar behaviors have been marked as number of events per group (e/g). Particular behaviors of guinea pigs, were classified as: positive, negative and sexual, based on the available literature. Data were compared with the Mann–Whitney U test, probabilities of less than 0.05 were considered statistically significant. Result of the present study showed that frequent exchange of enriching items did not influenced the frequency of positive behaviors like locomotion and popcorn. Social sniffing, also classified as positive, was twice as common in the lower frequency group (228 vs. 101 e/g, p < 0.05). Negative behaviors as biting, attacking and threatening intensified significantly in the group with higher frequency of items exchange, amounting 4 vs. 10, 3 vs. 13 and 7 vs. 23 e/g (p < 0.05), respectively. Based on our results we can conclude that too frequent rotation of enrichments items produces an increase of negative behaviors and reduces some positive behaviors of laboratory guinea pigs. 35 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Is the fur chewing in animals caused by Slitrk1 gene mutation? Iwona Guja, Stanisław Łapiński, Piotr Niedbała Department of Swine and Small Animal Husbandry, University of Agriculture in Krakow, Poland e-mail: iwona.guja@ur.krakow.pl Research on fur chewing in animals are carried out for years and did not explain the reason of occurring this obsessive compulsive spectrum disorder so far. Scientists all over the world try to discover what underlies the problem, which affects almost all vertebrate species. There are few genetic animal models of obsessive- compulsive disorders (OCD), like Hoxb8 knockout mice which manifests phenotype similar to trichotillomania or Slitrk5 knockout model which reveals OCD-like behavior, including anxiety and selfgrooming. Recent studies on Slitrk1 gene carried out on patients with OCD shows few mutations which could have impact in the disease development. Zuchner et al. (2006) characterized two non-synonymous sequence changes – G > A transition resulted in substitution of arginine for lysine (c.1751G>A, R584K) and replacement of serine by glycine (c.1777A > G, S593G). Ozomaro et al. (2013) described three mutations – L63L, T416S and N400I. First, L63L, does 36 not change the coding protein, but might have functional effect such as decreasing translation or impact on posttranslational modifications of protein. Second mutation, T416S, probably occurs in all people population without OCD and is functionally tolerated. The most interesting seems to be the third mutation, N400I, which is the missense change where asparagine is replaced by isoleucine. The functional effect of this mutation was proved in rat hippocampal neurons and mice cortical neurons (impact on neurite growth, which is stimulated by wild type Slitrk1). The most important is fact that both – Slitrk1 gene sequence and N400I mutations are conservative and occurs in many species. It gives a chance to repeat the research carried out by Ozomoro and Zuchner teams on animals in which OC spectrum disorders are observed, for example chinchillas. There would be a big chance to explain and understand a basis of fur chewing. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of feeding technology on calf behaviour Andrzej Kaczor1, Urszula Kaczor2 1 Department of Technology, Ecology and Economics of Animal Production, The National Research Institute of Animal Production, Balice near Krakow, Poland 2 Department of Animal Biotechnology, University of Agriculture in Krakow, Poland e-mail: andrzej.kaczor@izoo.krakow.pl Rearing of calves is one of the most difficult stages of cattle husbandry. The management system and also the scale of production largely determine the way milk or liquid feed is administered to calves. The introduction of automatic calf feeders enabled feeding calves in a group with an individual liquid feed ration formulated to meet their needs. Ethological studies provide valuable cognitive information about the usefulness of certain technological solutions from the perspective of animal comfort. The objective of the study was to determine the effect of management system and liquid feeding method on the behaviour of calves. The study was conducted with 30 Holstein-Friesian calves (Black-and-White variety) between 5 and 70 days of age. The control group (K) of 10 female calves were kept exclusively in individual igloo hutches with outside pens and fed 3 times daily with liquid feed from nipple buckets. The experimental group (D) of 20 female calves were kept in individual igloo hutches and then (until day 70) in group pens in an open-type calf shed, where they were fed from automatic calf feeders at all times. In the sixth week of life, 24-hour behavioural video observations were conducted to record lying (resting), standing and walking activities as well as liquid feed and concentrate intake. Frequency of feeder visits was also determined. During that period, animals received 6 kg of liquid feed and concentrate feed ad libitum. The results were statistically analysed with SAS ver. 10. The mean 24-h lying time of calves was 1005.6 min in group D (group pen) and 1055.2 min in group K. Calves from group D spent significantly more time standing and walking per 24 h (357.6 min) than did calves from group K (320.4 min) (P ≤ 0.05). The duration of concentrate feeding was 37.6 min/day in group D, being significantly shorter than in group K (52.4 min) (P ≤ 0.01). Calves from group D spent significantly more time on liquid feed intake (43.2 min) compared to those from group K (12.0 min) (P ≤ 0.01). The mean frequency of feeder visits was 12 times per day (range of 4 to 36 times). The management and liquid feeding systems did not significantly affect the total resting time of calves per day, but had a significant effect on differences in the duration of other activities. The greater range of movement, staying in a group and more frequent feeder visits in group D pen have increased standing and walking times as well as liquid feeding time during 24 h. Whereas shorter concentrate intake time was related to the management system, because group-housed animals compete for feed and consume it more quickly. 37 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of watering of cows in the milking parlour on their welfare Andrzej Kaczor1, Urszula Kaczor2 1 Department of Technology, Ecology and Economics of Animal Production, The National Research Institute of Animal Production, Balice near Krakow, Poland 2 Department of Animal Biotechnology, University of Agriculture in Krakow, Poland e-mail: andrzej.kaczor@izoo.krakow.pl During milking, cows lose considerable amounts of water with milk. This could lead to water deprivation, which causes discomfort in animals, especially those milked in parlours with no drinking bowls in the stalls. Cows that stand idle when being milked, focus their attention, among others, on udder handling and undesired movements. The aim of the study was to determine the effect of using drinking bowls in milking parlour on the behaviour of cows during milking and on water intake. The study was conducted with Holstein-Friesian cows of Black-and-White variety with daily yield of 25–35 kg milk. Cows were milked in a 2 × 6 herringbone milking parlour. Drinking bowls were placed in the milking stalls. The control group (K) were 76 cows milked in the parlour with no drinking bowls, and the experimental group (D) consisted of the same cows after placing the bowls in the milking parlour. Ethological studies were performed with video camera 1 day before and 30 days after the animals were watered during milking in the summer period. The study accounted for restless milking behaviours such as stepping and kicking the milking unit. Frequency and amount of water intake during milking was also 38 accounted for. The results were statistically analysed using Statistica ver. 9, based on one-way analysis variance. The total stepping behaviour of 76 cows during afternoon milking on day 30 of watering in the milking parlour (62 stepping events) was significantly lower than in cows milked on the day before the water bowls were placed in the parlour (89 stepping events) (P ≤ 0.01). Also Kicking of the milking unit by cows on day 30 of watering during milking also was much less frequent (4 kicking events) than in cows milked on the day before water bowl placement (12 kicking events). During milking the 76 cows drank water 73 times on day 30 after bowl placement. A single water intake by the cows was 8 sec and the amount of water intake was approx. 2 l/cow. The use of water bowls in the milking parlour had a positive effect on behaviour of cows in the parlour, thus improving their welfare. Giving animals access to water in the milking parlour to quench their thirst has made them less restless and nervous during milking. For this reason, the frequency of restless milking behaviour, such as stepping and kicking the milking unit has considerably decreased. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of slaughter age on physicochemical parameters of lamb meat Aldona Kawęcka, Michał Puchała, Anna Miksza-Cybulska Department of Animal Genetic Resources Conservation, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: aldona.kawecka@izoo.krakow.pl Wrzosówka is a native breed of pelt sheep with characteristic grey wool, high prolificacy and very good adaptation to harsh environmental conditions. Despite its low slaughter parameters, Wrzosówka lamb meat is considered a delicacy and many consumers compare it to game meat due to unique taste, aroma and colour of the meat. The objective of this study was to determine the effect of lamb slaughter age on physicochemical and organoleptic properties of meat obtained from Wrzosówka ram lambs. Six-month-old ram lambs were kept indoors under semi-intensive feeding conditions. Slaughter was performed on two dates: at 8 and 10 months of age. Raw leg muscle (m. biceps femoris) was analysed for pH of meat 24 and 48 h post mortem, drip loss from muscle sample during 24-storage, and L*, a* and b* coordinates 48 h postmortem using a Minolta CR 400. Roasted leg muscle was organoleptically evaluated for aroma, taste, juiciness and tenderness. The results of this study showed that slaughter age created significant differences in most of the physicochemical traits. pH measured after 48 h was higher (6.4 vs. 5.9), and drip loss was lower in the meat of older lambs. Meat from this group was darker (lower L* value) compared to that in the group of younger lambs (43.7 vs. 52.3), with a greater proportion of redness (higher a* value) – 20.0 vs. 16.8. The organoleptic evaluation of roasted leg showed significant differences in juiciness and tenderness. According to the testers, these parameters were more favourable for lamb meat obtained from younger animals. 39 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of housing system on slaughter traits of native Świniarka lambs Aldona Kawęcka, Jacek Sikora Department of Animal Genetic Resources Conservation, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: aldona.kawecka@izoo.krakow.pl The native breed of Świniarka sheep is very well adapted to local environmental conditions and is undemanding in feed. Świniarka sheep perform well in an extensive management system, on set-aside land, and are used with success for management of landscape, including environmentally valuable areas. Świniarka sheep provide very flavoursome meat, similar to game meat in taste. Lamb carcass known as Jagnięcina ze świniarki has been included in the List of Traditional Products. The purpose of the study was to compare meat performance of Świniarka ram lambs kept indoors and on pasture. The experiment was performed in Subcarpathia at a farm keeping native Świniarka sheep. Evaluation was made of 20 ram lambs aged 4 months, which were divided into two feeding groups. Ram lambs in the control group were kept in a sheep house; they received meadow hay and straw ad libitum as well as around 0.3 kg of crushed oats per lamb. In the second group, ram lambs remained on pasture throughout fattening and also received crushed oats. After reaching 7 months of age, the animals were 40 slaughtered and their carcasses subjected to slaughter analysis. At the start of the experiment, the body weight of 4-month-old lambs averaged 16.5 kg. At the end of the experiment (3 months later), the body weight was higher in the group of pastured lambs at 26.2 kg. Lambs reared indoors were 1.5 kg lighter. Dressing percentage was 38.2% in the control group and 39.2% in the pastured group, with a non-significant difference. The pastured group was superior for most of the carcass measurements except for carcass and loin length, which were greater in the confined lambs. The proportion of valuable cuts in the carcasses of ram lambs from both groups averaged 41%. Slaughter analysis showed that the lamb carcasses differed in the weight of cuts. The weight of leg, chump and shoulder was higher for the pastured group with highly significant differences. Also the meat content was significantly higher for pastured lambs at 993.3 g compared to 836.0 g in the control group. It is concluded that pasture management of Świniarka lambs had a positive effect on the slaughter parameters. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Meat quality in lambs of native breed fed the diet supplemented with maize DDGS Aldona Kawęcka, Ewa Sosin-Bzducha Department of Animal Genetic Resources Conservation, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: aldona.kawecka@izoo.krakow.pl The rapid development of biofuel production has increased the availability of plant processing by products, mainly rapeseed meal and cake, as well as dried distiller’s grains (DDGS). Due to their high nutritive value and relatively low price, these feeds are considered an excellent component of concentrate mixtures for farm animals. Maize DDGS is most widely used in sheep nutrition and it is characterized by a high content of crude protein, including a high proportion of ruminal bypass protein, as well as a high energy value. DDGS can successfully replace both energy and protein components in the concentrate mixture, and due to a high content of polyunsaturated fatty acids it can also modify fatty acid profile of the meat. The objective of the experiment was to determine meat quality of Wrzosówka lambs receiving a concentrate diet containing DDGS. The control group received a standard diet based on cereal components and soybean meal, and in the experimental group soybean meal and part of barley were replaced with DDGS. The fattening was carried out for 60 days (up to 8 months of age). Chemical composition was determined based on the samples of longissimus dorsi muscle. The composition of fatty acids was analysed with a VARIAN 3400 gas chromatograph. The meat of native Wrzosówka lambs fed the DDGS diet was characterized by a significant increase in the fatty acids content of C18:2 n-6 (9.73 vs. 6.59%, respectively) and CLA (1.59 vs. 0.58%, respectively). The meat of control lambs was found to contain more of linolenic acid C18:3 (n-3), which was reflected in a higher proportion of n-3 PUFA in the overall fatty acid pool. The introduction of DDGS into the concentrates had a beneficial effect on the CLA content but also caused a deterioration in n-6/n-3 PUFA in relation to the control group (13.61 vs. 7.06). Our study shows that the presence of DDGS in lamb concentrates allows modifying the fatty acid profile of meat fat. 41 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The impact of environmental enrichment on exploratory and hunting behavior of laboratory and companion animals Agata Kokocińska Institute of Genetics and Animal Breeding of the Polish Academy of Sciences, Jastrzębiec, Poland e-mail: a.kokocinska@ighz.pl Environmental enrichment significantly prevents from various kinds of behavioral disorders, such as obsessive – compulsive disorder, stereotypy, depression or anxiety and phobias. Exploratory behavior is a basic need of all animals, and inability to provide it, prevents the fulfillment of one of the five freedoms, namely the freedom to express normal behavior. Despite the confined space, which is limited to companion and laboratory animals by the human, this needs can be satiated to a certain extent. The best type of enrichments for macrosmatic animals (such is the vast 42 majority of laboratory and companion animals) is olfactory stimuli, stimulating the sense of smell. In addition, we distinguish a lot of environmental enrichments using the desire to gain the food (exploration and hunting behavior) which require a logical action (mental stimulation) and those that involve animal physically. Such solutions have a positive impact on the behavior of animals and prevent to psychosis, that in the case of laboratory animals should be considered an indirect effect of additional elements on the test results. Wpływ wzbogaceń środowiska na zachowania eksploracyjne i łowieckie zwierząt laboratoryjnych i towarzyszących Agata Kokocińska Instytut Genetyki i Hodowli Zwierząt Polskiej Akademii Nauk, Jastrzębiec e-mail: a.kokocinska@ighz.pl Wzbogacenia środowiska w znacznym stopniu zapobiegają różnego rodzaju zaburzeniom behawioralnym, takim jak zaburzenia obsesyjno-kompulsywne, stereotypie, depresja czy też lęki i fobie. Zachowania eksploracyjne są podstawową potrzebą wszystkich zwierząt. Brak możliwości jej zaspokojenia uniemożliwia osiągnięcie jednej z pięciu wolności, czyli wolności do wyrażania naturalnego zachowania. Mimo zamkniętej przestrzeni, którą człowiek stwarza zwierzętom towarzyszącym i laboratoryjnym, można tę potrzebę w pewnym stopniu zaspokoić. Najlepszym typem urozmaiceń dla zwierząt makrosmatycznych (a do takich należy zdecydowana większość zwierząt laboratoryjnych i towarzyszących) są bodźce zapachowe, stymulujące węch. Ponadto można wyróżnić całą gamę urozmaiceń środowiska wykorzystujących chęć zdobycia pożywienia (eksploracja, zachowania łowieckie), wymagających logicznego działania (stymulacja psychiczna), oraz takie, które angażują zwierzę fizycznie. Tego typu rozwiązania wpływają pozytywnie na behawior zwierząt i zapobiegają psychozom. Jednak w przypadku zwierząt laboratoryjnych należy mieć na uwadze pewien pośredni wpływ dodatkowych elementów na wyniki badań. 43 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Behaviour of Chinchilla lanigera male kept under farm conditions – preliminary studies Stanisław Łapiński1, Iwona Guja1, Katarzyna Mrozicka1, Piotr Niedbała1, Viktor A. Ilyukha2 Department of Swine and Small Animal Husbandry, University of Agriculture in Krakow, Poland Institute of Biology, Karelian Research Centre, Russian Academy of Science, Petrozavodsk, Russia 1 2 e-mail: s.lapinski@ur.krakow.pl Chinchilla breeding in Poland has 60 years of tradition. During this time, not only the scale of production, but also technology and attitude to animals have changed. Nowadays welfare is one of the most important aspects in modern animal production. Breeders need to take care not only of good feeding, health and housing area but also mental comfort. It is necessary to keep high level of welfare. Modern fur animals breeding should develop in the way of adapting production technologies to improve quality of animals life. The presented study concerned the behavior of chinchilla male in the polygamous breeding system. Chinchillas were kept on two farms with different experience in family life of animals (2 months and 2 years). In both farms the behavior of males was observed in the variant with access or no access to an individual cage. Using data from video recording, ethograms were prepared in order to analyse daily activities for each individual, different farms or breeding system (with or without individual cage 44 for a male). It was determined that neither farm nor breeding system had any influence on chinchillas’ daily activities. It was observed that male chinchillas spent 80% of their time resting and only 20% intaking food and water or other activities. Their activity levels increased during evening and night hours, and decreased during the day. Whether they had access to individual cages or not, male chinchillas spent most of time in the corridor. In older and more mature families, where animals were more familiar with each other, male chinchillas spent less time in the corridor (approximately 50%) than males in younger families (approximately 88%). The experiment proved that male chinchillas with unlimited access to their individual cages spent only 10% of the time in there. This proves that access to individual cage for a male in polygamous system is not required for improving welfare of chinchillas. Financed by DS-ZHTChiDI 3266/2015. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The differences in the biometric dimensions of the Silesian horses depending on the share of other breeds blood in their pedigrees Jarosław Łuszczyński, Kinga Dobrzańska, Magdalena Pieszka, Weronika Pisarczyk, Bogusława Długosz, Romana Augustyn, Zenon Podstawski, Monika Stefaniuk-Szmukier Department of Poultry Breeding, Institute of Animal Science, University of Agriculture in Krakow, Poland e-mail: m.pieszka@ur.krakow.pl The aim of the study was to analyze the biometric dimensions of Silesian mares and stallions depending on the participation of blood of other breeds in their pedigrees. We analyzed 838 silesian horses (459 mares, 379 stallions) registered in the I volume of VII section of the Stud Book of Silesian Horses. Depending on the percentage share of Oldenburg and Thoroughbred horses blood in pedigrees studied horses were divided into 6 groups. It was found that in the study population 52.9% mares and 57.7% stallions had in their pedigrees infusion of other breeds blood. The height at withers among mares and stallions with infusion of Oldenburg and Thoroughbred blood was highly significantly and significantly higher compared respectively to mares with 12.5% of Oldenburg blood and Silesian stallions and mares without the addition of blood of other breeds. Together with increasing of blood of Oldenburg and Thoroughbred horses decreasing of girth circumference among Silesian stallions was noted. The largest girth circumference was found in Silesian stallions without the share of foreign blood what was highly significantly and significantly different in comparison to other groups of stallions. The lowest cannon circumference was characterized for Silesian mares and stallions with 12.5–50% share of Thoroughbred blood. According to the exterior evaluation the highest notes were obtained by mares with 12.5–75% share of Thoroughbred and Oldenburg blood and stallions with 25% share of Oldenburg blood g which highly significantly and significantly different compared to other analyzed groups of horses. The criteria given in the breed pattern for horses of old type was met by mares without any share of foreign blood or only 25% Oldenburg blood share as well as stallions without any share of foreign blood, with Oldenburg blood or stallions with share of Oldenburg and Thoroughbred blood together. Horses in new type were represented only by the Silesian mares with pedigrees containing more than 25% of the Oldenburg blood, in the case of stallions none of the groups did not meet the established requirements. 45 Różnice w wymiarach biometrycznych koni śląskich w zależności od udziału w ich rodowodach krwi koni innych ras Jarosław Łuszczyński, Kinga Dobrzańska, Magdalena Pieszka, Weronika Pisarczyk, Bogusława Długosz, Romana Augustyn, Zenon Podstawski, Monika Stefaniuk-Szmukier Zakład Hodowli Koni, Instytut Nauk o Zwierzętach, Uniwersytet Rolniczy w Krakowie e-mail: m.pieszka@ur.krakow.pl Celem pracy była analiza wymiarów biometrycznych klaczy i ogierów śląskich w zależności od udziału w rodowodach krwi koni innych ras. Analizie poddano 838 koni śląskich (459 klaczy, 379 ogierów) wpisanych do I działu VII tomu Księgi Stadnej Koni Śląskich. W zależności od procentowego udziału krwi koni oldenburskich i pełnej krwi angielskiej w rodowodach koni śląskich, klacze i ogiery tej rasy podzielono na 6 grup. Stwierdzono, że w badanej populacji 52,9% klaczy oraz 57,7% ogierów miało w rodowodach dolew krwi koni innych ras. Wzrost klaczy i ogierów z dolewem krwi oldenburskiej oraz pełnej krwi angielskiej okazał się wysoce istotnie i istotnie większy w porównaniu odpowiednio do klaczy z 12,5% dolewem krwi oldenburskiej i do ogierów i klaczy śląskich bez dolewu obcej krwi. Wraz ze zwiększającym się dolewem krwi koni oldenburskich i pełnej krwi angielskiej zmniejszał się obwód klatki piersiowej ogierów śląskich. Największy obwód klatki piersiowej stwierdzono u ogierów śląskich bez dolewu obcej krwi, co wysoce istotnie i istotnie róż- 46 niło się w porównaniu z pozostałymi grupami ogierów. Najmniejszym obwodem nadpęcia cechowały się zarówno klacze, jak i ogiery śląskie, u których w rodowodach stwierdzono udział koni pełnej krwi angielskiej na poziomie 12,5–50%. Pod względem pokroju najwyżej oceniono klacze z 12,5–75% dolewem krwi oldenburskiej i pełnej krwi angielskiej oraz ogiery z 25% dolewem krwi oldenburskiej, co wysoce istotnie i istotnie różniło się w porównaniu z innymi grupami badanych koni. Kryteria podane we wzorcu rasowym dla koni w starym typie spełniały klacze bez dolewu obcej krwi lub z dolewem krwi oldenburskiej nie większym niż 25% oraz ogiery bez dolewu obcej krwi, z krwią oldenburską lub ogiery z jednoczesnym dolewem krwi oldenburskiej i pełnej krwi angielskiej. Konie w nowym typie były reprezentowane tylko przez klacze śląskie, w rodowodach których płynęło więcej niż 25% krwi oldenburskiej, w przypadku ogierów żadna z grup nie spełniała ustalonych wymagań. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Influence of timely gutting fish on intensity of infection of muscle tissue by the Anisakis larvae Elena L. Mikulich The Belarusian State Agricultural Academy, Gorki, Belarus e-mail: helenamikulich1971@gmail.com The Republic of Belarus has not a sea access and imports marine fish mainly from the Atlantic, Arctic and Pacific. The fish are mostly stored frozen (about 80%). The fish inhabit the natural fisheries can be infected by various types of parasites e.g. nematodes of genus Anisakis (Dujardin 1845). The most common representative of this genus is A. simplex, which larvae infect the internal organs and skeletal musculature of host. The aim of this work was to study of the intensity of the salmon muscle infection by A. simplex larvae in dependent to term and manner of gutting. The frozen carcass of pink salmon (weight 820– 970 g), five ungutted and ten gutted undecapitated fish, were examined parasitologically. The Anisakis larvae were found in all analyzed carcasses. However, they were found more often in ungutted fish. The largest number of larvae was detected in the muscle tissue of abdominal walls (in a single case even 135 pieces), on the surface of the abdominal cavity and internal organs (5–9 pieces) and few (1–2 pieces) in the head end of the muscles of the abdominal wall. Quite another picture of infection intensity was in the gutted salmons. There were found 10–16 parasites on the fish. This can be explained the fish are emptied as soon as possible and less larvae can penetrate from the abdomen through into the muscles. The obtained data confirm opinion that post-harvest Anisakis larvae migrate from the body cavity in contiguous muscle tissue, resulting in lost value of the raw materials. 47 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The use of PCR and real-time PCR for qualitative and quantitative determination of horse meat Małgorzata Natonek-Wiśniewska, Piotr Krzyścin Department of Animal Genomics and Molecular Biology, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: malgorzata.natonek@izoo.krakow.pl The authenticity of the food is now a serious problem for scientists, consumers and the food industry at all levels of the production process. In this sector, there is time to counterfeiting or unfair supplementation products not complying with the declaration. EU rules very strictly and regulate in detail the issues which necessitates the development of techniques that can catch the prohibited practice. In recent years, attention has been paid to the possibility of using molecular methods to identify the species because of their high sensitivity and specificity, as well as short and low costs of analysis. The aim of the work undertaken to develop a sensitive and specific method of qualitative and quantitative determination of the component horse contained in meat and dairy products. 48 Qualitative identification method was developed using PCR technology, and the TaqMan probe qPCR and were used to identify quantitative. The study was carried out with DNA from horse meat at 100%, 12%, 1.5%, 0.5%, 0.05%. Detection and quantification limits are 0.005% and 0.05%. The results obtained for quantitation of independent isolation of DNA from the same samples are reproducible (RSDcT ≤ 0.21%). Designated concentrations are accurate (for c ≥ 0.05 DC is ≤ 5.01%). The method enables the determination of DNA horse food, regardless of the degree of processing of the sample. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Concentration of heavy metals in increments of common carp’s scales (Cyprinus carpio L.) Patryk Nowacki, Ewa Łuszczek-Trojnar, Ewa Drąg-Kozak, Magdalena Socha, Włodzimierz Popek Department of Ichthyobiology and Fisheries, University of Agriculture in Krakow, Poland e-mail: e.trojnar@ur.krakow.pl The aim of the study was to determine the concentration of heavy metals Cu, Cd, Zn, Ni, Mn, Fe and Pb in scales of carp coming from two different fish farms: Experimental Station in Mydlniki supplied with the Rudawa River and private carp farm in Ludynia supplied with underground water source, and located at area used for hunting. An additional goal was to verify thesis that the level of metals in scales is a permanent indication of environmental contamination. After the decapitation of 3-year-old carp all their scales were collected for heavy metals analysis. Some of scales were left as the whole, but the rest of them were divided precisely into three parts accord- ing to annual growth rings. Additionally the hepatopancreas and kidney were taken for metal level analysis. In all samples the concentration of studied metals were determined by atomic absorption method using the spectrometer ATI UNICAM 929. There were significant differences between concentration of metals in the scales of carp coming from Ludynia and from Mydlniki. Statistically significant differences between metal contents in particular increments of scales were observed in fish coming from both farms. Obtained results confirm usefulness of fish scales as the indicator of environmental contamination with heavy metals. 49 Koncentracja metali ciężkich w pierścieniach przyrostów rocznych łusek trzyletnich karpi (Cyprinus carpio L.) Patryk Nowacki, Ewa Łuszczek-Trojnar, Ewa Drąg-Kozak, Magdalena Socha, Włodzimierz Popek Katedra Ichtiobiologii i Rybactwa, Uniwersytet Rolniczy w Krakowie e-mail: e.trojnar@ur.krakow.pl Celem pracy było określenie koncentracji metali ciężkich, takich jak Cu, Cd, Zn, Ni, Mn, Fe i Pb, w łuskach karpi pochodzących z dwóch gospodarstw rybackich: z Rybackiej Stacji Doświadczalnej UR w Mydlnikach, której stawy są zasilane wodą z rzeki Rudawy, i z prywatnego gospodarstwa karpiowego w Ludyni, którego stawy zasilane z podziemnych źródeł znajdują się na terenach wykorzystywanych do polowań. Dodatkowym celem tej pracy była próba weryfikacji tezy, że poziom metali w łuskach jest trwałym odzwierciedleniem stanu skażenia środowiska. Od trzyletnich karpi pobrano wszystkie łuski, z których część pozostawiono w całości, natomiast pozostałe podzielono 50 precyzyjnie według pierścieni przyrostów rocznych na 3 części. Poziom metali określono metodą AAS przy użyciu spektrometru ATI UNICAM 929. Wykazano istotne różnice w poziomie stężenia metali w łuskach pomiędzy grupami karpi z gospodarstwa w Ludyni i w Mydlnikach. Analiza statystyczna wykazała też istotne różnice średnich koncentracji metali w poszczególnych pierścieniach przyrostów rocznych łusek ryb z obu gospodarstw. Metale ciężkie trwale akumulują się w pierścieniach przyrostów rocznych łusek, co potwierdza przydatność tej tkanki jako wskaźnika zanieczyszczenia środowiska metalami ciężkimi. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The freezing point of milk of Polish Holstein-Friesian cows Agnieszka Otwinowska-Mindur, Ewa Ptak Department of Genetics and Animal Breeding, University of Agriculture in Krakow, Poland e-mail: rzmindur@cyf-kr.edu.pl The objective of this study was to estimate the influence of milk composition on the freezing point of first-lactation milk of Polish Holstein-Friesian cows. Data comprised 1,612,419 milk samples from the first lactations of 313,744 Polish Holstein Friesian cows and were made available by the Polish Federation of Cattle Breeders and Dairy Farmers. The cows calved in 19,402 herds in 2013 and 2014. Six-factor analysis of variance using the GLM procedure in SAS was applied to milk freezing point. The following factors were included in the analysis: daily milk yield (up to 16 kg, 16–24 kg, > 24 kg), fat content (up to 3.6%, 3.6–4.4%, > 4.4%), protein content (up to 3.1%, 3.1–3.5%, > 3.5%), urea content (up to 150 mg/L, 150–270 mg/L, > 270 mg/L), lactose content (up to 4.75%, 4.75–5.00%, > 5.00%) and somatic cell count (up to 400,000 in mL, > 400,000 in mL). The mean freezing point of milk (–0.5351ºC) – and more than 95% of all milk samples – did not exceed the quality limit for the freezing point of cow’s raw milk, which, following Polish standards, was set at –0.52ºC. The effects of all examined milk components on the freezing point of milk (fat, protein, urea and lactose content, somatic cell count) were highly significant (P < 0.001). Only daily milk yield class did not affect it significantly (P = 0.0662). On average, the freezing point was highest for lower daily milk yield and for lower fat, protein, urea and lactose content. Milk with somatic cell counts over 400,000 had a slightly higher freezing point (–0.5337ºC) than milk qualified to the “extra” class, i.e. up to 400,000 somatic cells in mL (–0.5354ºC). 51 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Concentration of harmful gas components in organic sheep houses according to the season Paweł Paraponiak Department of Technology, Ecology and Economics of Animal Production, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: pawel.paraponiak@izoo.krakow.pl The objective of the study was to evaluate the concentration of harmful gases in organic sheep houses during the summer and winter seasons. Above-normal concentrations of harmful gas components, especially in the winter period, could have a direct negative effect on animal productivity and on the quality of products obtained. The concentration of CO2, NH3 and H2S was recorded (Dräger Multiwarn 2 gas analyser) in four organic sheep houses (hall-type construction, deep litter, gravity ventilation, normal stocking density) in 6 measurement series. In all the facilities, no presence of the most harmful compound (H2S) was found in both the summer and winter periods. CO2 content varied considerably between 0.04 and 0.20% (P ≤ 0.01), as did NH3 concentration, which ranged from 1 to 8 ppm (P ≤ 0.05). The measurements were 52 significantly higher in the winter period (0.11‒0.20% and 4‒8 ppm, respectively) than in the summer period (0.04‒0.14% and 1‒6 ppm, respectively), which was due to weaker air circulation during this period. A concentration map of different gases in the buildings showed that their highest accumulation was in the central part and near the walls deprived of inlet ventilation holes, whereas in the regions nearer the walls with ventilation holes their concentration was lower. In conclusion, the concentration of the analysed gases did not exceed the acceptable limits, and most of the measurements, especially those taken in summer, showed much lower levels, which indicates that stocking density, flooring quality and efficiency of the ventilation system inside the house were appropriate. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Variation in lamb behaviour according to the housing system Paweł Paraponiak Department of Technology, Ecology and Economics of Animal Production, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: pawel.paraponiak@izoo.krakow.pl The aim of the study was to determine the effect of housing lambs in open and semi-open systems on their behaviour. Behaviour was recorded and analysed for a total of 60 Świniarka ram-lambs of uniform age. Equal sized groups (20 animals per group) were kept in three different housing systems: open on pasture (O; permanent pasture with access to overhead shelter, permanent grazing system); open rotational grazing on pasture (K; permanent pasture, variable rotational grazing system); and semi-open backyard [P; animals kept on green area (not a pasture) with free 24-hour access to the sheep house]. The results of the study show considerable (even 10%) differences in the basic types of observed sheep behaviour depending on the housing system (P ≤ 0.01). In general, the large pasture area along with greater distribution of feed (forage) stimulated the higher walking activity of ram-lambs from groups O and K in terms of pasture exploration – movement of animals and feed intake itself (25.0, 31.9% and 24.1, 34.2%, respectively). With the increasing pasture area and the simultaneously decreasing proportion of the most palatable plants (legumes and herbs), animals were engaged in greater walking activity, spreading into a larger area in search of the most palatable food, leaving as a result part of the sward ungrazed (this was considerably limited under the rotational system). The lower walking activity of lambs from group P (19.3%, P ≤ 0.01) resulted from the smaller outdoor area available (which in practice mainly extended the living space of animals but in terms of fodder base its contribution was marginal) and above all the easier access to roughage deposited in feeders translated into both shorter eating time (24.0%) and relatively smaller mobility of the animals. As a result, animals from the semi-open system were engaged in less walking activity and spent considerably more time lying down and standing (36.4 and 20.3%, respectively, P ≤ 0.01). 53 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Characterisation of the population of border collie dogs breed in Poland Magdalena Pieszka, Anna Halek, Jarosław Łuszczyński, Bogusława Długosz, Romana Augustyn, Monika Stefaniuk-Szmukier, Zenon Podstawski, Weronika Pisarczyk Department of Horse Breeding, Institute of Animal Science, University of Agriculture in Krakow, Poland e-mail: m.pieszka@ur.krakow.pl Border collie, dogs bred in the UK to herding sheep, well find themselves also in canine sports and field and rubble rescue. Amazing is their help for people with disabilities or with depressive disorders. Border collie, both in the utility and exhibition type, are increasingly popular around the world. In our country most of the breeding is based on obtaining a dog with a beautiful conformation and correct body structure, free from genetic diseases. The first dogs of this breed were imported into Poland in 1996, since then, the number of dogs increases together with the number of kennels, there are now more than 100. In 2014, the population of dogs registered by the Polish Kennel Club has increased compared to 2009, almost double the largest increase was seen in the number of bitches (2009: 271, 2014: 511) and breeding bitches (2009: 93, 2014: 206). According to statistics, the number of litters increased from 37 to 82 and also puppies 54 born: the number of males born in 2014 was twice as high (229) than in 2009 (114). Also the number of females since 2009 (108) increased to 209, but in the last two years, it remained at similar level. Among the provinces with the largest number of representatives of border collie following can be distinguished: Wielkopolskie, Dolnośląskie, Śląskie, Mazowieckie and Małopolskie. Most breeding females were recorded in the province Mazowieckie (96), sires in Śląskie (18 pcs.) and the maximum number of litters was found in Dolnośląskie (16 litters). Kennels having the largest number of dogs of this breed are: Energizer, Alderaan, Never Never Land, Q’Celtic, Forest Land Chilabo, Alchera, Legendary Star Dust and Viva La Tia. The most beautiful dogs in the exhibition were Alderaan PALO ALTO, Trucharm MY Brillantino, Locheil I’M THE DIVA and Tehya DESTINY OF HER OWN. Charakterystyka populacji psów rasy border collie w Polsce Magdalena Pieszka, Anna Halek, Jarosław Łuszczyński, Bogusława Długosz, Romana Augustyn, Monika Stefaniuk-Szmukier, Zenon Podstawski, Weronika Pisarczyk Zakład Hodowli Koni, Instytut Nauk o Zwierzętach, Uniwersytet Rolniczy w Krakowie e-mail: m.pieszka@ur.krakow.pl Border collie, psy wyhodowane w Wielkiej Brytanii do pasienia owiec, świetnie odnajdują się też w sportach kynologicznych oraz w ratownictwie terenowym i gruzowym. Zadziwiająca jest także ich pomoc osobom niepełnosprawnym lub z zaburzeniami depresyjnymi. Psy rasy border collie, zarówno w typie użytkowym, jak i wystawowym, są coraz bardziej popularne na całym świecie. W Polsce większość hodowli opiera się na uzyskaniu psów o pięknym eksterierze i prawidłowej budowie ciała, wolnych od chorób genetycznych. Pierwsze psy tej rasy sprowadzono do Polski w 1996 roku. Od tego czasu zwiększa się nie tylko ich liczba, ale również liczebność hodowli, których obecnie jest już ponad 100. W 2014 roku populacja psów rejestrowanych przez ZKwP wzrosła w porównaniu do roku 2009 prawie dwukrotnie, przy czym największy wzrost był widoczny w liczbie suk (2009: 271 szt., 2014: 511 szt.) i suk hodowlanych (2009: 93 szt., 2014: 206 szt.). Według statystyk, z 37 do 82 wzrosła liczba miotów oraz szczeniąt w nich urodzonych: liczba urodzonych samców w 2014 roku była dwukrotnie wyższa (229 szt.) niż w 2009 roku (114 szt.). Także liczba suczek od 2009 roku (108 szt.) zwiększyła się do 209 szt., jednak w ostatnich dwóch latach utrzymywała się ona na podobnym poziomie. Wśród województw z największą liczbą przedstawicieli rasy border collie można wyróżnić: wielkopolskie, dolnośląskie, śląskie, mazowieckie oraz małopolskie. Najwięcej suk hodowlanych odnotowano w woj. mazowieckim (96 szt.), reproduktorów w woj. śląskim (18 szt.), a najwyższa liczba miotów została stwierdzona w woj. dolnośląskim (16 miotów). Hodowle posiadające największą liczbę psów tej rasy to: Energizer, Alderaan, Never Never Land, Q’Celtic, Forest Land Chilabo, Alchera, Legendary Star Dust oraz Viva La Tia. Najbardziej utytułowane psy wystawowe to Alderaan PALO ALTO, Trucharm MY BRILLANTINO, Locheil I’M THE DIVA oraz Tehya DESTINY OF HER OWN. 55 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Characteristics of work of chosen hippotherapy centres in southern Poland Zenon Podstawski, Justyna Kądzior, Bogusława Długosz, Romana Augustyn, Monika Stefaniuk-Szmukier, Magdalena Pieszka, Jarosław Łuszczyński, Weronika Pisarczyk Department of Horse Breeding, Institute of Animal Science, University of Agriculture in Krakow, Poland e-mail: z.podstawski@ur.krakow.pl One of the most popular methods used in the therapy of children is treatment involving horses. While hippotherapy has been known for many years, the availability of the literature and research on the subject is still very limited. The horse has served the man in different ways. In addition to the use for military, transportation and agriculture purposes, we recognized the therapeutic relevance of this animal. The impact of hippotherapy, established through a continuous dialogue between the patient and the horse’s movement, almost always is properly coupled with the psychological aspect – communication with the living creature. Through direct contact with the body of the horse the sensormotor sphere is activated, allowing the feeling of heat, 56 odors, body awareness, current position in space, planning movements. The variety of texture in the construction of the outer horse hair softness, roughness mane and tail, to a large extent contribute to the proper sphere of sensory stimulation. The aim of the study was to analyse the activity and organization of centres of Horse Assisted Therapy. The analysis included the activity of four randomly selected centres. Based on interviews and observations results demonstrated that: They were wellorganized institutions, the purpose of their activities was primarily to help disabled patients, all centres were established timetables, employed highly specialized staff. Charakterystyka działalności wybranych ośrodków hipoterapeutycznych w Polsce południowej Zenon Podstawski, Justyna Kądzior, Bogusława Długosz, Romana Augustyn, Monika Stefaniuk-Szmukier, Magdalena Pieszka, Jarosław Łuszczyński, Weronika Pisarczyk Zakład Hodowli Koni, Instytut Nauk o Zwierzętach, Uniwersytet Rolniczy w Krakowie e-mail: z.podstawski@ur.krakow.pl Jedną z coraz popularniejszych metod stosowanych w terapii dzieci są zajęcia z udziałem koni. Chociaż hipoterapia jest znana od wielu lat, dostępność do literatury oraz badań na ten temat bywa wciąż bardzo ograniczona. Od zarania dziejów człowiek wykorzystywał konie. Koń służył i pomagał człowiekowi na różne sposoby. Oprócz wykorzystywania do celów militarnych, transportu osób i ładunków czy pracy na roli, bardzo wcześnie dostrzeżono również znaczenie terapeutyczne tego zwierzęcia. Oddziaływanie hipoterapii, odbywające się poprzez ciągły dialog pomiędzy pacjentem a koniem, jego ruchem, praktycznie zawsze jest odpowiednio sprzężone z aspektem psychologicznym – komunikacją z żywą istotą. Poprzez bezpośredni kontakt z ciałem konia aktywizowana zostaje sfera sensomotoryczna oznaczająca odczuwanie ciepła, zapachów, świadomość ciała, jego aktualnego położenia w przestrzeni, planowania ruchów. Różnorodność faktury w budowie zewnętrznej konia, miękkość sierści, szorstkość grzywy i ogona, w dużym stopniu sprzyjają właściwej stymulacji sfery czuciowej. Celem pracy była analiza działalności i organizacja ośrodków prowadzących działalność z zakresu hipoterapii. Analizą objęto działalność czterech, losowo wybranych ośrodków. Na podstawie przeprowadzonych wywiadów i obserwacji własnych wykazano, że: ośrodki były dobrze zorganizowanymi instytucjami, celem ich działania była przede wszystkim pomoc pacjentom, wszystkie ośrodki miały ustalone plany zajęć, zatrudniano wyspecjalizowaną kadrę psychologiczną, lekarsko-terapeutyczną. 57 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The effect of bacteria addition on the chemical and microbiological composition of the whole plant corn silage Jan B. Pyś1, Zygmunt M. Kowalski1, Marta Malkiewicz1, Jadwiga Flaga1, Łukasz Korytkowski1, Vanessa Demey2 1 Department of Animal Nutrition and Dietetics, University of Agriculture in Krakow, Poland 2 Lallemand Animal Nutrition, Blagnac, France e-mail: rzpys@cyf-kr.edu.pl During this study, the whole maize was ensiled (Pioneer 9400, FAO 270, dry matter 419,7 g ∙ kg–1, particles length 8–10 mm) and the following silages were prepared: C0 – without addition of bacterial inoculant and CI – CIII with addition of bacterial inoculant (1g of preparation). Inoculants used in the study contained respectively: CI – Lactobacillus plantarum 6.0 × 104, Pediococcus acidilactici 3 × 103, P. pentosaceus 1 × 104; CII – L. plantarum 2.0 × 105, P. acidilactici 1 × 105, P. pentosaceus 1 × 105, Propionibacterium acidipropionici; CIII – P. acidilactici 1 × 105, P. acidipropionici 1.5 × 105 CFU∙ g of fresh forage. The lowest (P < 0.05) DM (402.9 g ∙ kg–1) was found in the silage CIII, while in the other silages the DM content equaled: C0 – 409.9, CI – 410.4 and CII – 58 408.3 g ∙ kg–1. Bacterial additives used in the study did not affect ash, crude protein, NDF and ADF content in all silages, but decreased (P < 0.05) water soluble carbohydrates in CI – 2.3, CII – 1.3 and CIII – 1.1 g ∙ kg–1 DM silage, in comparison to the control (C0 – 3.8 g ∙ kg–1). After opening of the container, the amount of lactic acid bacteria differed significantly (P < 0.05) between silages and equaled as follows: C0 – 4.9065, CI – 3.9164, CII – 5.2829 and CIII – 4.6013 log10CFU ∙ g–1 of fresh matter. The highest (P < 0.05) amount of yeast was found in CIII silage (4.9607), comparing to the other silages: C0 (4.5694), CI (4.5195) and CII (4.7556) log10CFU ∙ g–1 of fresh matter. Molds, in the amount of 0.8000 log10CFU ∙ g–1 of fresh matter, was isolated only from C0 silage. Table 1. Chemical and microbiological composition of whole plant maize silage Item C0 CI CII CIII Dry matter g · kg–1 409.9b 408.3b 414.2a 402.9c g · kg–1 of dry matter Crude ash 39.0a 40.3a 36.7b 37.5ab Crude protein 81.2 79,0 79,5 79.6 NDF 319.7b 327.1a 316.9b 315.1b ADF 192.3 192.0 192.9 193.3 WSC 3.8a 1.3a 1.1a 1.1a log10 jtk · kg–1 of fresh matter a, b, c, d LAB 4.9065b 5.2892a 4.4046c 3.9164d Yeast 4.5694b 4.7556a 3.9543c 4.51195b Molds 0.8000b 0.0000b 0.0000b 0.0000b – numbers marked with different letters differ significantly P < 0.05. 59 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The effect of bacteria addition on the fermentation quality and aerobic stability of the whole plant corn silage Jan B. Pyś1, Zygmunt M. Kowalski1, Marta Malkiewicz1, Jadwiga Flaga1, Łukasz Korytkowski1, Vanessa Demey2 1 Department of Animal Nutrition and Dietetics, University of Agriculture in Krakow, Poland 2 Lallemand Animal Nutrition, Blagnac, France e-mail: rzpys@cyf-kr.edu.pl During this study, the whole maize was ensiled (Pioneer 9400, FAO 270, dry matter 419.7 g ∙ kg–1, particles length 8–10 mm) and the following silages were prepared: C0 – without addition of bacterial inoculant and CI – CIII with addition of bacterial inoculant (1 g of preparation). Inoculants used in the study contained respectively: CI – Lactobacillus plantarum 6.0 × 104, Pediococcus acidilactici 3 × 103, P. pentosaceus 1 × 104; CII – L. plantarum 2.0 × 105, P. acidilactici 1 × 105, P. pentosaceus 1 × 105, Propionibacterium acidipropionici; CIII – P. acidilactici 1 × 105, P. acidipropionici 1.5 × 105 CFU ∙ g of fresh forage. The lowest (P < 0.05) DM content (402.9 g ∙ kg–1) was found 60 in the silage CIII. Bacterial additives used in the study did not affect the pH and NDF content in silages. The lowest (P < 0.05) amount of N-NH3 (59.8 g ∙ kg–1 of total-N) and ethanol (19.9 g ∙ kg–1 DM) was found in silage CIII. Used additives increased (P < 0.05) lactic acid content (62.4 g ∙ kg–1 DM) and reduced (P < 0.05) acetic acid content (14.8 g ∙ kg–1 DM) in the silage CIII, comparing to the concentration of this acids in the silage C0 (54.4 i 29.1 g ∙ kg–1 DM, respectively). The aerobic stability period of CI, CII and CIII silages was significantly longer (P < 0.05) than that of the C0 silage (121 h) and equaled respectively: 168, 162 and 141 h. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Changes in some fat parameters during storage of butter made from the milk of cows under two feeding systems Iwona Radkowska Department of Technology, Ecology and Economics of Animal Production, The National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: iwona.radkowska@izoo.krakow.pl The main determinant of the technological process of butter making and of the quality and shelf life of the end product is the quality milk. Raw material quality is largely determined by feeding method and housing conditions of the cows, their health, and the hygienic quality of milk. The aim of the study was to evaluate parameters of butter made from the milk of cows under two feeding systems: TMR (group I) and pasture (group II). During control milkings, milk for butter making was collected three times per season from 10 cows in each group. The properly prepared cream was churned in a batch-type butter churn for 45‒60 minutes until the granules were the size of 2‒3 mm. After separation of buttermilk, butter granules were rinsed 3 times with water at 11°C, and then kneaded until water droplets were evenly distributed. Butter was analysed after 1, 15 and 30 days of storage. The basic composition, acid value, peroxide value, TBA, iodine value, fat melting point and fat solidification point were determined at the Department of Animal Product Processing of the University of Agriculture in Cracow. Butter made from the milk of cows from group II was characterized by significantly (p ≤ 0.05) higher acid number, peroxide value, TBA, and iodine number. Peroxide value was slightly exceeded in fat samples after 15-day storage of butter from group II (0.36 meq oxygen ∙ kg‒1 fat) and after 30-day storage of butter from group I (0.35 meq oxygen ∙ kg‒1 fat). It is accepted that oxidative rancid smell and flavour are perceptible with peroxide values above 1. An important quality criterion of milk fat is the proportion of unsaturated fatty acids expressed as iodine number. Butter made from the milk of cows from group II was characterized by higher iodine number at every stage of storage. The unsaturation index of milk fat is the main factor determining its nutritive and healthpromoting value, as well as butter consistency. In the present study, butter from group II had significantly (p ≤ 0.05) lower melting and solidification temperature. Excessively hard butter consistency, which often occurs during the winter feeding of cows, is caused, among others, by the low content of unsaturated fatty acids. Fat melting and solidification points illustrate changes in fat consistency depending on temperature, and butter spreadability depends on their value. 61 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Vitamin and cholesterol content of milk depending on type of feeding Iwona Radkowska1, Adam Radkowski2 Department of Technology, Ecology and Economics of Animal Production, National Research Institute of Animal Production, Balice near Krakow, Poland 2 Department of Grassland Management, Institute of Plant Production, University of Agriculture in Krakow, Poland 1 e-mail: iwona.radkowska@izoo.krakow.pl Milk is an important and well-studied source of fatsoluble vitamins, in particular vitamins A and E. The concentration of these vitamins in milk depends on various factors, including feeding season, management conditions, stage of lactation, herd yield, udder health, and genetic factors. Feeding cows with green roughages has a positive effect on increasing the content of vitamins A, E, C and b-carotene in milk. The aim of the study was to determine the effect of Simmental dairy cow feeding system on the vitamin A and E and cholesterol content of milk. Cows were divided into three experimental groups. Cows in group I were kept in a barn and fed TMR, cows in group II were pastured for approximately 6 h and fed in a barn with TMR (half the diet of cows from group I), and cows in group III were kept on pasture and their feeding was based on pasture sward. The experiment was conducted from May to October. During control milkings, milk for chemical analyses was collected three times per season from 10 cows in each group. Vitamins A and E were determined according to SOP M.001 methodology ver. 6 of 19 March 2012 “Simultaneous determination of vitamins A and E in 62 solid or bulk materials”, and cholesterol according to SOP M.023a ver. 1 of 20 October 2011 “GC determination of cholesterol in meat and milk”. Vitamin A content was the lowest in group I (0.36 µg ∙ ml‒1), followed by 0.40 ug ∙ ml‒1 in group II and significantly higher (p ≤ 0.05) in group III (0.50 ug ∙ ml‒1). Similar relationships were observed for vitamin E. The lowest content was found in group I (1.01 ug ∙ ml‒1), followed by 1.16 ug ∙ ml‒1 in group II and 1.39 ug ∙ ml‒1 in group 3. Milk cholesterol content showed considerable variation. Significantly (p ≤ 0.05) the lowest content was found in the milk of cows from group III (13.86 mg ∙ 100 g‒1), followed by 15.84 mg ∙ 100 g‒1 in group II and the highest (p ≤ 0.05) cholesterol content in the milk of cows from group I (17.71 mg ∙ 100 g–1). In summary, it is concluded that pasture feeding of cows has a positive effect on the milk vitamin content and contributes to reducing milk cholesterol. Fresh pasture sward is characterized by higher levels of vitamin E and provitamin A compared to conserved feeds, which is why the milk of grazed cows contains more vitamins than the milk of cows fed TMR. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of foliar fertilization with silicon on quality and nutritive value of meadow sward silages Adam Radkowski1, Iwona Radkowska2 Department of Grassland Management, Institute of Plant Production, University of Agriculture in Krakow, Poland 2 Department of Technology, Ecology and Economics of Animal Production, The National Research Institute of Animal Production, Balice near Krakow, Poland 1 e-mail: a.radkowski@ur.krakow.pl An important criterion for evaluating the quality of roughages for ruminants is the energy and protein value. Feed which are given to animals should meet the requirement for maintenance and production. In turn, a major concern in plant production is to reduce the effects of adverse environmental factors, especially drought periods. These negative effects can be attenuated by using preparations that stimulate processes in plants, which allow for activating stresscoping mechanisms. Silicon is a component that stimulates the growth and development of plants. This element is not considered as the components necessary for the life of plants, or to macronutrients or micronutrients, however, it is in significant quantities in most of the plants (especially in monocotyledons) and this amount is comparable with the contents of calcium, magnesium and phosphorus. Silicon reduces the toxic effect of excess manganese and iron, positive effects on the ionic balance in plants. It strengthens the cell walls, increases the resistance of plants to fungal diseases and adverse environment, including very low temperature and water stress. However, an excessive amount of silicon decreases the digestibility of plants. This relationship is important with respect to the grass, which are used as the only feed in animal nutrition. Species of the grass family are characterized by differences in silicon content. The objective of the study was to determine the effect of foliar fertilization of meadow sward with silicon on nutritive value of silage as well as quality and basic chemical composition of milk. Analysis of the silage for chemical composition showed that higher crude protein content and lower crude fibre content were obtained for silages made from meadow sward fertilized foliarly with silicon compared to control silages. The silage obtained from silicon-fertilized plants was characterized by higher PDIN and PDIE values as well as higher energy value (UFL, UFV). Foliar fertilization of meadow vegetation with silicon had a positive effect on cow productivity and improved the microbiological quality of milk. 63 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Quality and nutritive value of silages made from selected grass-legume mixtures Adam Radkowski1, Iwona Radkowska2 Department of Grassland Management, Institute of Plant Production, University of Agriculture in Krakow, Poland 2 Department of Technology, Ecology and Economics of Animal Production, The National Research Institute of Animal Production, Balice near Krakow, Poland 1 e-mail: a.radkowski@ur.krakow.pl The aim of the study was to compare the chemical composition and nutritive value of three types of clover-grass mixtures differing in the proportion of red clover (10, 20 and 40%) and perennial meadow sward as a control object. The field trial was performed in 2013–2015 in a private farm. The experiment was conducted on acid brown soil (pHKCl 5.2) of class V. Each plot had an area of 1200 m2. The experimental silages were prepared from first cut. The material was harvested in two stages, first the forage was cut with a rotary mower at the heading-flowering stage, and then it was dried by a single tedding. The dry matter content ranged 35–40%. After six weeks from preparation of silages were started feeding. Before feeding, representative samples for chemical analyses were taken from silages to determine the content of basic components according to the Weende method, pH on a pH meter, and ammonia level using Conway’s method. The organic acids content was determined by gas chromatography. Nutritive value was estimated with INRA units using Winwar software. 64 The grass-legume mixtures were characterized by significantly higher content of organic and mineral components compared to the control mixture. The crude protein and crude fat content of the silages from grass-legume mixtures (average for these three objects) was higher by 16 and 21%, respectively, and the content of P, K, Ca, Mg and Na was higher (average values for the three objects grass-legume mixtures) by 12, 15, 19, 13 and 14%, respectively, compared to the control. The silages contained (in 1kg of DM) in terms of energy value from 0.77 to 0.92 UFL and from 0.68 to 0.83 UFV and the protein value from 82 to 98 for PDIN and from 80 to 93 for PDIE. The results show that 20 and 40% participation of red clover significantly improves the nutritional value of the obtained silages. The nutritional value will depend on the species composition of sward because not all plants have the same value as forage. In agricultural practice should be greater use for the production of silages grassy-clover sward. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Genetic relationship between conformation traits and lactose percentage and urea concentration in milk of Polish Holstein-Friesian cows Alicja Satoła, Ewa Ptak, Wojciech Jagusiak Department of Genetics and Animal Breeding, University of Agriculture in Krakow, Poland e-mail: a.satola@ur.krakow.pl The objective of this study was to estimate the genetic correlations of lactose percentage and urea concentration in milk with conformation traits related to udder and legs of Polish Holstein-Friesian cows. Data consisted of 5.813 test-day records and type scores of 791 primiparous cows. Two descriptive traits (udder, feet and legs, scored from 50 to 100) and 11 linearly scored traits (describing udder: fore udder height, rear udder height, central ligament, udder depth, udder width, fore teat placement, teat length, rear teat placement; describing legs: rear legs – side view, foot angle, rear legs – rear view, on a scale of 1 to 9) were included in the analysis. Genetic correlations were calculated based on (co)variances estimated using the Bayesian method via Gibbs sampling and the multitrait animal model. Genetic correlations between lactose content and conformation traits ranged from –0.18 to 0.23. Genetic correlations between milk urea concentration and conformation traits ranged from 0.02 to 0.39. Average genetic correlations with daily lactose percentage were higher than 0.15 (ignoring sign) only for udder (descriptive trait) and a few linearly scored traits: central ligament, udder depth, rear teat placement, and rear legs – rear view. Milk urea content was weakly or moderately genetically correlated with six type traits: udder, and five linearly scored traits: fore udder height, central ligament, udder width, teat length, and rear legs – side view. Genetic correlations between these traits were higher than 0.15 (ignoring sign). Our results showed that type traits connected with udder were more highly correlated with both lactose and milk urea content than leg conformation traits. Selection for improvement of legs will not affect lactose percentage and milk urea content, but increases in both lactose percentage and milk urea concentration may be expected as an indirect response to selection for better udder. 65 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of breed and aging time on physicochemical and organoleptic quality of beef Ewa Sosin-Bzducha, Michał Puchała Department of Animal Genetic Resources Conservation, The National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: ewa.sosin@izoo.krakow.pl The experiment used the meat of 16 Polish Red-andWhite and Simmental bulls slaughtered at the age of 24 months. Bulls received maize silage and hay or ensiled hay supplemented with concentrate. Animals were kept in breed groups in a free-stall building on straw, and had constant access to water. Physicochemical and organoleptic analyses were carried out on meat (m. thoracis, n = 64) aged for 1, 7, 14 and 21 days in vaccum bags. Analysis of physicochemical properties include changes in pH, loss during ripening, the heating loss, color (CIE Lab system) and shear force (V-blade, Warner-Bratzler). Meat with a different degree of maturity undergo thermal treatment was evaluated by a sensory changes in the intensity and specificity of odor and taste, as well as discoloration of meat, juiciness and tenderness. Statistical analysis was performed with Statistica ver. 9.1 66 software (2009). The meat from the Polish Red-andWhite breed was characterized by more yellow colour (5.96 vs. 4.79) and distinctly higher chroma (21.93 vs. 21.06) compared to Simmental meat. The meat from the conserved breed was more tender (69.28 vs. 93.61 N). During aging, lightening of muscle colour, increased redness and chroma, and improved tenderness were observed. High scores were given for aroma intensity and taste desirability. The highest scores were awarded to meat aged for 7 and 14 days. The evaluation showed that meat tenderness and delicate texture steadily improved until day 14, after which they remained at a similar level or slightly deteriorated after 21 days of aging. Meat from the conserved breed generally had better scores, which could be influenced by better component scores for delicate texture and juiciness, as well as taste intensity. Wpływ rasy i czasu dojrzewania na jakość fizykochemiczną i organoleptyczną wołowiny Ewa Sosin-Bzducha, Michał Puchała Dział Ochrony Zasobów Genetycznych Zwierząt, Instytut Zootechniki PIB, Balice k. Krakowa, Polska e-mail: ewa.sosin@izoo.krakow.pl Materiał doświadczalny stanowiło mięso pochodzące od 16 buhajków rasy polskiej czerwono-białej i simentalskiej, ubitych w wieku 24 miesięcy. Buhajki żywione były standardowo, kiszonką z kukurydzy i sianem lub sianokiszonką uzupełnianą paszą treściwą. Zwierzęta utrzymywano w grupach rasowych, alkierzowo, systemem wolnostanowiskowym na słomie ze stałym dostępem do wody. Analiza fizykochemiczna i organoleptyczna została przeprowadzona na mięsie (m. thoracis, n = 64) dojrzewającym w workach próżniowych przez 1, 7, 14 i 21 dni. Analiza cech fizykochemicznych obejmowała zmiany pH, ubytku podczas dojrzewania, ubytku termicznego, barwy (system CIE Lab) oraz siły cięcia (V-blade, Warnera-Bratzler). Mięso o różnym stopniu dojrzałości poddane obróbce termicznej oceniano sensorycznie pod względem zmian intensywności i swoistości smaku oraz zapachu, a także zmian barwy mięsa, soczystości i delikatności. Analizę statystyczną przeprowadzono za pomocą programu Statistica ver. 9,1 (2009). Mięso pochodzące od rasy polskiej czerwono-białej charakteryzowało się bardziej żółtą barwą (5,96 vs. 4,79) oraz zdecydowanie większym natężeniem barwy (21,93 vs. 21,06) w stosunku do mięsa pochodzącego od rasy simentalskiej. Mięso pochodzące od rasy zachowawczej charakteryzowało się większą kruchością (69,28 vs. 93,61 N). W trakcie dojrzewania obserwowano rozjaśnienie barwy, wzrost wysycenia w kierunku czerwieni i zwiększenie natężenia barwy, a także poprawę kruchości mięsa. Wysoko oceniono intensywność zapachu mięsa oraz pożądalność smaku. Najlepiej oceniano mięso dojrzewające przez 7 i 14 dni. Ocena wykazała poprawę kruchości i delikatności mięsa następującą stopniowo do 14. dnia, a następnie utrzymanie parametru na zbliżonym poziomie lub nieznaczny spadek jego oceny po 21 dniach dojrzewania. Mięso pochodzące od rasy zachowawczej zostało lepiej ocenione ogólnie, na co wpływ mogły wywrzeć lepsze oceny cząstkowe w zakresie delikatności i soczystości, a także intensywności smaku. 67 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of litter size on mortality of new born cavies’ youngs Petr Tejml, Miloslav Šoch Faculty of Agriculture, University of South Bohemia in České Budějovice, Czech Republic e-mail: tejml@seznam.cz These days breeding of guinea pig pets is very popular in Europe and other continents. It has a big potential to the future, because guinea pigs can be kept and bred also in town flats. Cavia sows have in one litter from one to six youngs. The embryonal sacs are very solid and have to be ruptured by their mothers. This is one of big problems in a cavy breeding, because due to it up to 40% of youngs are suffocated during the birth. The aim of this study was to find out the effect of litter size on mortality new born youngs. The moni- 68 toring was carried out in four cavy studs of hobby cavies with different animal hygiene conditions and various type of breeding technology. From the obtained results we can see that mortality of new born youngs was statistically significant in small and large litters. The lowest mortality was proved in middle size litters with three or four youngs. The factor of breeding technology did not have any influence on the mortality. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of herd size on implementation of the genetic resources conservation programme for Hucul horses Iwona Tomczyk-Wrona Department of Animal Genetic Resources Conservation, National Research Institute of Animal Production, Balice near Krakow, Poland email: iwona.wrona@izoo.krakow.pl Hucul is a primitive breed of horses, which means that their development has been strongly influenced by the natural environment. Because of this, the natural, stable and feeding conditions are of utmost importance to the breed, which is included in the genetic resources conservation programme. Essential for horses, which are a social species, is the size of the group in which they live. It is an important factor influencing not only the physical development, but, perhaps more importantly, the mental development of these animals. Polish and foreign authors emphasize the role of herd and its size on the ontogenic development of the horses. The possibility of living in a herd allows young horses to develop their herd instinct, which is particularly important for stallions. Where horses are only tethered under stable management, the visual and olfactory contact with more animals itself has a positive effect on maintaining the relatively normal mental state of the animals. The best solution would be to keep horses only in large herds, preferably in the open systems. Unfortunately, for economic reasons, the realities most often look completely different. Herd size and the possibility of living in a herd is particularly important for primitive horses such as Huculs. The first formal programme for conservation of the breed was developed in 2000 and provided for a limited number of 210 jobs (allocated each year in advance) for mares aged 1 year old and above, as well as for stallions owned by the herds included in the conservation programme. The payment scheme has changed since 2005. It was required to own at least three adult mares that have been entered into the studbook and selected for the conservation programme. Since 2008, the limit of mares has been decreased to 2 per herd. This met the expectations of herd owners applying for subsidy, but was detrimental to the horses and their breeding, where 5 brood mares per herd is considered the minimum for breeding and selection work. In 2005, 505 mares from 85 herds were selected for the conservation programme. Most herds (45.9%) had 4 to 10 mares, 41.2% herds had 3 mares, 5.9% herds had between 11 and 20 mares, and 3.5% of the herds had more than 20 mares. In 2015, 1225 mares in 264 herds were included in the Hucul horse conservation programme. This means that in 2015 the number of eligible mares increased 2.4-fold and the number of herds increased 3.1-fold compared to 2005. The number of mares in 69 the herds (2015) is as follows: 58.0% herds have 2 or 3 mares, 35.6% herds have from 4 to 10 mares, 4.9% herds have from 11 to 20 mares, and 1.5% herds have more than 20 brood mares. The number of herds with 4 to 10 mares increased from 45.9% in 2005 to 58.0% in 2015, but the proportion of herds with 11–20 and more than 20 mares has decreased. The increase in the population of Hucul horses participating in the conservation programme is a highly beneficial outcome of the conservation programme, but unfortunately it is not paralleled by an increase in the number of mares in the herd. The introduction of such a low 70 minimum number of two mares per herd caused that not only breeders, but also horse “keepers” joined the conservation programme, which is not good for the horses themselves. The incomplete herd impact coupled with inadequate knowledge of the methods of primitive Hucul horse breeding among the owners, may adversely influence the physical and mental development of the horses. Work done as part of the multiannual programme 17– 1.10.9. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of implementing the genetic resources conservation programme on the distribution of Hucul horses in Poland Iwona Tomczyk-Wrona Department of Animal Genetic Resources Conservation, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: iwona.wrona@izoo.krakow.pl Huculs, the small primitive mountain horses are considered one of the oldest Polish breeds with fixed genotype. The breed originated in the East Carpathian Mountains (Wooded Carpathians), in the upper reaches of the Cheremosh, Prut, Putylka, Moldova, Suceava and Tysa Rivers and in the area of Bukovina. They are descendants of various horse types: Tatar, Oriental, Arabian, Turkish, Przewalski’s, as well as horses with Noric blood. However, the exact origin of the Hucul breed is unknown until today. Importantly, the breed has developed under the influence of the environment – harsh mountain climate, scarce feed, and primitive living conditions. Their name comes from Hucul highlanders, people with unique culture for whom horses played a very important role in their daily lives. The first literary reference to Hucul horses dates from 1603 when Dorohostajski described them in “Hippika” [Equestrianism] as excellent mountain horses that perform very well in the harshest conditions. Since 1979, they have reached the status of a conservation breed and are considered as the national cultural and breeding heritage. Work on the domestic conservation programme for farm animal genetic resources was begun in Poland in 1999. In May 2000, the Ministry of Agriculture accepted the implementation of conservation programmes for different populations, including the Hucul horse population. The subsidy granted at that time as part of the biological fund covered only 210 jobs. The conditions changed in 2005, when the Hucul conservation programme was expanded to include 505 mares (32.3% from the Małopolskie province, 24.9% from the Podkarpackie province, 10.1% from the Śląskie and Opolskie provinces, and 8.5% from the Wielkopolskie province). The least Hucul mares (below 1%) were in the Dolnośląskie, Mazowieckie and Pomorskie provinces. Zachodnio-Pomorskie and Podlaskie were the only provinces with no Hucul horses participating in the conservation programme. The year 2015 has brought major changes, because the number of Hucul mares increased 2.4-fold to 1225 in relation to 2005. The distribution has also changed because now the Hucul horses are found in all the provinces and the number of mares under 71 the conservation programme is comparable in each province. Today most Hucul mares are found in the Podkarpackie pro-vince (25.9%) followed by the Małopolskie (20.6%), Lubelskie (10.0%), Łódzkie (7.1%), Wielkopolskie (6.3%), Warmińsko-Mazurskie (5.6%) and Mazowieckie (4.9%) provinces. It should be noted that the Hucul breed is perfectly compatible with conditions of the conservation programme. The fact 72 that Huculs are found in all regions of Poland is indicative of the excellent adaptability of the breed to various environmental conditions, from native mountain areas to lowlands. Work done as part of the multiannual programme 17– 1.10.9. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Polish collection and communication of inspection results system and food chain information system as tools for welfare improvement at slaughterhouse Michał Tracz, Agnieszka Jackowska-Tracz, Krzysztof Anusz Department of Food Hygiene and Public Health Protection, Warsaw University of Life Science, Poland e-mail: michal_tracz@sggw.pl Official veterinarians are essential part of a modern meat inspection model which important part deals with animal welfare protection. Task in area of welfare protection are divided between official veterinarians, food business operators and livestock producers. Exchange of information between these actors is important to support them in making inform decisions in area of animal welfare and health. Food chain information and collection and communication of inspection results (FCI/CCIR) are key points in modernization of meat inspection at a slaughterhouse level. FCI and CCIR were designed to support data exchange between all parties responsible for animal health and welfare. FCI is provided by livestock suppliers and consist of information on animals health and welfare status. CCIR is an important task of official veterinarians. Quality data provided by official veterinarians through CCIR supports animal health, welfare and food safety as well as environmental protection and sustainability. Aim of the study was to determine if Polish legal system and procedures of data exchange performed by official veterinarians at slaughterhouse are compliant to European Union legal requirements in area of animal welfare. Current Polish legal system do not fully facilitate the ability to share and use food chain information up and down the food chain. Analysis of polish legal requirements in area of FCI and CCIR leads to the conclusion that a formal system is compliant with the requirements of the law but in practice technical solutions do not fully enable the realization of the set goals of animal welfare protection. 73 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of ageing time on microstructure and tenderness of pig semimembranosus muscle Dorota Wojtysiak, Magdalena Górska, Joanna Wojciechowska Department of Animal Anatomy, University of Agriculture in Krakow, Poland e-mail: wojtysiakd@wp.pl One of the most important meat traits for consumer satisfaction is meat tenderness. Meat tenderness depends on several factors such as muscle fibre composition, sarcomere length, pH, intramuscular fat content and rate of tenderization. An important role in shaping meat tenderness is also played by connective tissue, which has been shown to be a critical factor in meat tenderness. Therefore, the aim of the current study was to investigate the effect of ageing time on muscle fibre traits, structural properties of intramuscular connective tissue and tenderness of m. semimembranosus in pigs. The study used 12 Polish Landrace × Polish Large White pigs slaughtered at 105 kg body weight. The muscle samples of m. semimembranosus were collected immediately after slaughter (45 min) and after storage of meat for 24 h, 96 h and 168 h at +4°C. Muscle samples were taken to categorize muscle fibre types (I, IIA and IIB) according to their NADH-TR activity and immunohistochemical determination of slow myosin heavy chain. The collagen architecture of 74 the muscles was examined under a scanning electron microscope JSM-5410. Fibre type percentage, fibre size and the following meat quality parameters, such as shear force and collagen content were determined. The current finding indicate that m. semimembranosus is a typical white muscle, rich in type IIB glycolytic fibers. Moreover, it was found that after 24 h storage the diameter of type I, IIA and IIB fibres decreases significantly. The analysis of intramuscular connective tissue under scanning electron microscopes demonstrated that, with ageing time, the arrangement of collagen fibrils in the perimysium become looser and less regular and form a loose network structure. In the case of meat quality parameters, no effect of meat storage time on the amount of total collagen was observed. In contrast, the amount of soluble collagen and shear force decreased gradually with time of storage. Funded by DS-3253. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The effect of Pedi bolus specimen on the improvement of cows’ hooves health Justyna Żychlińska-Buczek1, Edyta Bauer2, Andrzej Ochrem1, Elżbieta Sowula-Skrzyńska3, Grzegorz Skrzyński1 1 Department of Cattle Breeding, Institute of Animal Science, University of Agriculture in Krakow, Poland 2 Department of Genetics and Animal Breeding, University of Agriculture in Krakow, Poland 3 Department of Technology, Ecology and Economics of Animal Production, The National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: jzychlin@ar.krakow.pl The main goal of every dairy farmer is to achieve maximum profits. Issues related to the diseases of the hoof are nowadays one of the three main reasons for culling cows from the herd. What in addition generates supplementary costs of breeding. The aim of this thesis was to determine the effect of Pedi bolus specimen on the reduction the amount of lameness and improvement of selected reproductive indicators of dairy cows in the herd. The con- ducted research shows that the using of bolus Pedi has a beneficial effect on increasing hoofs health. Period between pregnancies for animals treated with Pedi bolus was satisfying, whereas ratio for OMW related to the calving period exceeded normative value (above 405 days). It has been reported a 14% effectiveness increase of the first fertilization and decrease about a half of the cows number requiring three-time insemination. 75 Wpływ preparatu Pedi bolus na poprawę zdrowotności racic krów Justyna Żychlińska-Buczek1, Edyta Bauer2, Andrzej Ochrem1, Elżbieta Sowula-Skrzyńska3, Grzegorz Skrzyński1 1 Zakład Hodowli Bydła, Instytut Nauk o Zwierzętach, Uniwersytet Rolniczy w Krakowie Katedra Genetyki i Metod Doskonalenia Zwierząt, Uniwersytet Rolniczy w Krakowie 3 Dział Technologii, Ekologii i Ekonomiki Produkcji Zwierzęcej, Instytut Zootechniki – Państwowy Instytut Badawczy, Balice k. Krakowa 2 e-mail: jzychlin@ar.krakow.pl Głównym celem każdego hodowcy bydła mlecznego jest osiągnięcie maksymalnego zysku. Problemy związane z chorobami racic to obecnie jeden z trzech powodów brakowania krów ze stada, co generuje dodatkowe koszty. Celem pracy było określenie wpływu preparatu Pedi bolus na zmniejszenie liczby kulawizn oraz poprawę wybranych wskaźników reprodukcyjnych krów mlecznych w stadzie. Z przeprowadzonych badań wynika, że preparat Pedi bolus wpływa 76 korzystnie na poprawę zdrowotności racic. Okres międzyciążowy u zwierząt, u których zastosowano preparat, kształtował się na zadowalającym poziomie, natomiast wartość okresu międzywycieleniowego (OMW) przekraczała wartości normatywne (powyżej 405 dni). Zanotowano 14-procentowy wzrost skuteczności pierwszego zapłodnienia. Natomiast o połowę zmniejszyła się liczba krów wymagająca trzykrotnego zabiegu inseminacji. Session 2 Biotechnology and animal models International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Fur-bearing animals as the models in biomedical research Viktor A. Ilyukha1, Svetlana N. Sergina1, Ekaterina P. Antonova1, Evgeny A. Khizhkin1, Lyudmila B. Uzenbaeva1, Aleksandra G. Kizhina1, Tatiana N. Ilyina1, Irina V. Baishnikova1, Stanisław Łapiński2, Piotr Niedbała2, Marcin W. Lis2, Oleg V. Trapezov3, Yongping Xu4 Institute of Biology, Karelian Research Centre, Russian Academy of Science, Petrozavodsk, Russia 2 Faculty of Animal Science, University of Agriculture in Krakow, Poland 3 Institute of Cytology and Genetics, Siberian Branch, Russian Academy of Sciences, Novosibirsk, Russia 4 Dalian University of Technology, China 1 e-mail: ilyukha@bio.krc.karelia.ru Restriction of crossing, gene drift, and a certain degree of inbreeding in the course of industrial domestication of the fur animals resulted in that many recessive mutations brought to specialized fur farms “under cover” of the standard phenotype from natural populations (where they emerged stochastically) have become homozygous and phenotypically expressed. These mutations have been demonstrated to have a wide spectrum of pleiotropic effects. In 2014 Adam S. Wilkins with colleagues proposed that the domestication syndrome results predominantly from mild neural crest cell deficits during embryonic development. Most of the modified traits, both morphological and physiological, can be readily explained as direct or indirect consequences of such deficiencies. Some of these modified traits in domesticated animals are very similar with signs of several human disorders and consequently might be used in biomedical research. For example, homozygous white animals such as Hedlund mink (hh) have symptoms similar to the Waardenburg syndrome; minks with the Aleutian coat color gene (a/a) and sapphire foxes (b/b p/p s/s) have characteristics associated with the Chédiak-Higashi syndrome. Moreover, it is possible to use the carnivorous furbearing animals as models in ecological research for studying applied ecological issues such as pollution, disease, and global climate change. Some of the characteristics of the mammals make them useful models for studying these environmental problems are their trophic importance and environmental sensitivity. The husbandry and life history of animals reared in captivity are well understood, and this has permitted controlled experiments to be conducted. The study was carried out under state order (projects № 0221-2014-0001) and it was partially funded by the President of the Russian Federation grant for Leading Scientific School #1410.2014.4 and partially of the grant for Russian-Polish Interacademic Cooperation. 79 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Transgenic mice in cancer research Alicja Jozkowicz Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Kraków, Poland email: alicja.jozkowicz@uj.edu.pl Insights into the molecular and cellular basis of cancer biology are to a large extent derived from small animal models, especially transgenic mice. The use of systemic or conditional and tissue-specific knock-out or knock-in gene targeting strategies, including the inducible and tissue-specific Cas9-dependent mutagenesis, have allowed the generation of loss-of-function or gain-of-function mutations in tumor suppressors or proto-oncogenes. Such models become a basic tool in studies on resistance to drug treatment, oncogene cooperation, or role of microenvironment in tumor growth and formation of metastasis. Moreover, lineage tracing has recently been used to explore the tumor heterogeneity and to analyze neoplastic stem cells and cancer cell subpopulation dynamics. Finally, the study of immune response to growing tumor can be refined by use of patient-derived xenograft in humanized mice, with immune system reconstituted by human hematopoietic cells, or by conditional cell depletion in mice expressing a diphtheria toxin receptor under the control of cell-type specific promoter. Importantly, modification of even minor parameter may significantly influence the model used. In 80 the simplest approach, most frequently applied in the screening of anticancer drug effectiveness, the suspension of cancer cells is injected subcutaneously into mouse. Instead, we have proposed the subcutaneous injection of a single spheroid. Direct comparison of the tumor development from the same number of cells delivered as a cell-suspension or cell-spheroid revealed that the latter grew significantly slower, but were better vascularized and less necrotic. Moreover, they were more heavily infiltrated by mesenchymal stromal cells and hematopoietic progenitor cells, and contained more cells with cancer initiating cell phenotype. Accordingly, spheroid model is the more suitable and sensitive tool for analysis of anticancer drugs which target tumor angiogenesis or cancer stem cells or tumor microenvironment. Thus, mouse models are indispensible for understanding the tumor biology and can be very helpful in translational studies of anticancer therapies. However, all of them have some limitations, and can be sensitive to even modest changes in experimental protocols. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Influence endurance workout on selected physiological parameters of Arabian horses Romana Augustyn1, A. Pustuła1, Bogusława Długosz1, Tomasz Próchniak2, Magdalena Pieszka1, Jarosław Łuszczyński1, Weronika Pisarczyk1, Monika Stefaniuk-Szmukier1, Zenon Podstawski1 1 Institute of Animal Science, Department of Poultry Breeding, University of Agriculture in Krakow, Poland 2 Department of Biological Basis of Animal Production, University of Life Sciences in Lublin, Poland e-mail: r.augustyn@ur.krakow.pl This paper examines the influence of the pressures exerted on the endurance races horses. The research was conducted on Arabian horses of different ages and different training advancement. The aim of this study was to investigate whether gradually increasing intensity of effort during the training season affects the level of selected indicators of blood and physiological parameters of Arabian endurance horses. 3 blood collection were conducted, at each a material of horses at rest, right after exercise and 30 min after the end of exercise was tested. Moreover, after each training session the heart rate, movement and dehydration were examined. The number of haematocrit, leukocytes, erythrocytes and platelets, level of haemoglobin, value of MCV, MCHC, contains of to- tal plasma protein (CK) and creatine kinase were determined. The results indicated that horses were good preparing for the beginning of the season and in the vast majority their blood parameters were contained within the reference values. The level of total plasma protein and haematocrit indicate to the good state of water and electrolyte management and small variations CK determined for insignificant destructive effects of exercise on the horses muscle tissue. Strong individual characteristics of obtained results shows that only year-round research and determination of individual horse predisposition will allow to precise determination of progress in training and adequate reaction from the horse body on the effort. 81 Wpływ treningu rajdowego na wybrane parametry fizjologiczne koni czystej krwi arabskiej Romana Augustyn1, A. Pustuła1, Bogusława Długosz1, Tomasz Próchniak2, Magdalena Pieszka1, Jarosław Łuszczyński1, Weronika Pisarczyk1, Monika Stefaniuk-Szmukier1, Zenon Podstawski1 Instytut Nauk o Zwierzętach, Zakład Hodowli Koni, Uniwersytet Rolniczy w Krakowie Katedra Biologicznych Podstaw Produkcji Zwierzęcej, Uniwersytet Przyrodniczy w Lublinie 1 2 e-mail: r.augustyn@ur.krakow.pl W pracy zbadano wpływ obciążeń, jakim poddane zostały konie startujące w sportowych rajdach konnych. W doświadczeniu wzięły udział konie czystej krwi arabskiej w różnym wieku i w różnym zaawansowaniu treningowym. Celem pracy było zbadanie, czy stopniowo wzrastająca intensywność wysiłku w czasie sezonu treningowego wpływa na poziom wybranych wskaźników krwi oraz parametrów fizjologicznych koni rajdowych. Przeprowadzono 3 pobrania. Podczas każdego zbadano krew koni w spoczynku, zaraz po wysiłku i 30 min po zakończeniu wysiłku. Ponadto, po każdym treningu zbadano tętno, ruch oraz odwodnienie. Oceniano liczbę hematokrytową, liczbę leukocytów, erytrocytów i płytek krwi, poziom hemoglobiny, wartość MCV, MCHC, zawartość białka całkowitego oraz kinazy kreatynowej. Uzyskane wyniki wskazywały na dobre przygotowanie koni do sezonu startowego oraz – w zdecydowanej większości – mieściły się w normach referencyjnych. Poziom 82 białka całkowitego i hematokrytu wskazywał na dobry stan gospodarki wodno-elektrolitowej, a małe wahania powysiłkowego CK stanowiły o mało destrukcyjnym wpływie wysiłku na tkankę mięśniową koni. Trening rajdowy (wysiłek długotrwały) spowodował wzrost spoczynkowych wartości płytek krwi oraz krwinek białych. Początek sezonu treningowego charakteryzował się największym wahaniem zawartości białka całkowitego, a wraz z kolejnymi treningami różnice te były znacznie mniejsze. Badane konie wykazywały się zadowalającym przygotowaniem kondycyjnym do startów w zawodach, czego dowodzi szybki powrót analizowanych parametrów krwi do normy po wysiłku. Silnie osobniczy charakter uzyskanych wartości świadczy o tym, że dopiero całoroczne badania i ustalenie indywidualnych predyspozycji konia pozwoli na precyzyjne określenie postępów w treningu i adekwatnej reakcji ze strony organizmu konia na wysiłek. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Experimental gender related obesity effect of diet Agnieszka Baranowska1, Beata Skowron1, Katarzyna Ciesielczyk1, Justyna Domagała2, Piotr J. Thor1 1 Department of Pathophysiology, Jagiellonian University Medical College, Krakow, Poland 2 Department of Microbiology, Jagiellonian University Medical College, Krakow, Poland e-mail: abaranowska@cm-uj.krakow.pl Introduction: High-calorie diet is responsible for excessive weight gain. Obesity has recently become world epidemics, affecting not only adults but also children, which makes it the biggest health problem in the world. Yet the underlying mechanism remains a matter of debate. Objectives: The aim of this study was to clarify the role of gender in high fat diet induced obesity in pups and adult animals. Materials and methods: Female rats were fed low/ high fat diet during mating, pregnancy and lactation. The offspring and adult rats fed different diet had their body weight and temperature measurements taken twice a week. On the 21st day of the experiment the animals underwent anesthesia in order to have their blood samples collected for lipid profile. Results: After 3 weeks on HF diet female pups body weight was higher than in control group (p < 0.05). Contrary to the female pups, the increase in body weight was higher (p < 0.05) in male pups and occurred after 2 and 3 weeks. In adult female rats body weight increased after 2 weeks on HF, while in adult male group such weight gain was observed no sooner than after 3 weeks. After three weeks of the experiment body weight was correlated positively (r = 0.941) with lipid profile of adult both gender groups on HF diet. Conclusions: In male pups group body weight increased faster and achieved higher values then in female pups. On the contrary, in adult group of females body weight increased faster than in male rats and achieved similar values. 83 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Method of positioning the catheter into cisterna magna and fourth ventricle of the brain in sheep Aneta Bocheńska1, Agnieszka Pietsch-Fulbiszewska1, Natalia Sowińska1, Katarzyna Kirsz2 1 Center for Experimental and Innovative Medicine, University Center for Veterinary Medicine UJ-UR, Krakow, Poland 2 Department of Animal Biotechnology, University of Agriculture in Krakow, Poland e-mail: a.pietsch-fulbiszewska@ur.krakow.pl Drugs infusion through a catheter into the cisterna magna (CM) and repeated sampling of the cerebrospinal fluid (CSF) allow to determine the impact of chemicals on the central nervous system without passing drugs through the peripheral circulation and blood-brain barrier. Developing an effective, reproducible and minimally invasive method of collecting CSF is particularly useful for exploring central nervous system disorders and study the interactions of the nervous and endocrine systems. Surgical field included area between the occipital protuberance to the spinous process of the fourth cervical vertebra, the width of the wings of the first cervical vertebra. The animal was placed on the right side. A catheter was introduced into atlanto – occipital space to the cisterna magna. Puncture site was determined by median line and intracranial-dorsal edge of the wings of the atlas, caudally to the external occipital protuberance. The set of disposable Uniever combined epidural anesthesia minitray (polyurethane catheters, radiopaque, 90 mm, 18G) was used. Puncture at an angle of approx. 75–80 degrees, nasally, towards to the cisterna magna was performed. The 84 needle was inserted into the mid-length. When the CSF appeared through the guide – needle the catheter was placed, and then the needle was removed. The free end of the catheter was sutured to the skin of the animal and secured by dressing. The control of the location of the catheter was carried out by fluoroscopic examination and computed tomography. In the bone window, in the sagittal cross-section of the three-dimensional reconstruction portion of the catheter was visualized in the cerebrospinal reservoir and the tip was in the lateral recess of the fourth ventricle of the brain. The present modified method of injection into the CM is accurate, replicable, and safe. It might be applicable for assessing the pharmacological effects and mechanisms of the central action of drugs on the nervous system in sheep. Research supported by DS 3242/KBZ/2015 (Method of positioning the catheter into cisterna magna and fourth ventricle of the brain in sheep). International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Intrаbreed variability of the Ukrainian Mountain Carpathian sheep genetic structure under using of the different types of molecular genetic markers Taras Chоkаn1, Anna Radko2, Serhiy Tarasjuk3, Agnieszka Szumiec2, Dominika Rubiś2 Institute of Animal Biology NAAS, Lviv, Ukraine National Research Institute of Animal Production, Balice near Krakow, Poland 3 Institute of Fisheries NAAS, Kyiv, Ukraine 1 2 e-mail: tchokan@ukr.net The genetic structure of the population is determined mainly by the diversity of the gene pool, which includes both the general properties of species and genetic features that have arisen under adaptation of the population to the specific conditions of its existence. Thus the analysis of the genetic structure of Mountain Carpathian sheep with different color wool, its monitoring using different types of molecular genetic markers is the issue of vital importance. The research of genetic structures using different types of molecular genetic markers (DNA markers – from 11 microsatellite DNA sequences recommended by the FAO for assessing the biodiversity of sheep and genetic and biochemical systems – 13 loci). Microsatellite (STR) in most cases, are characterized by species conservatism, but the analysis revealed a number of markers specific by close groups. In total 106 alleles were detected, the number of which ranges from 3 (locus MAF65) in sheep with white wool to 12 (locus INRA63) in sheep with gray wool. The high polymorphic information content (PIC) it was averaged 0.740 in groups of sheep with gray wool and ranged from 0.525 (locus Oar304) to 0.851 (locus HSC). In our studies, the highest polymorphism was typical for loci INRA063 in sheep with black wool, which consisted of 13 alleles while the PIC was 0,865 and the observed heterozygosity (HO) – 0.878. The results of studies on the genetic structure of genetic and biochemical systems indicate the presence of intrabreed genetic differentiation. The data analysis proved that certain genetic and biochemical systems, particularly Tf and PN, can be used as markers for specific characteristics of the genetic structure of the studied groups of animals. For the distribution of allelic variants of genes and genotypes studied groups differ from one another, which of course is the result of different factors of natural and artificial selection and different approaches in the process of breeding them. 85 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The influence of orexin B on spontaneous and GnRH-stimulated LH secretion from dispersed pituitary cells of female Prussian carp Jarosław Chyb, Gabriela Cora, Piotr Lisowski, Magdalena Socha, Mirosława Sokołowska-Mikołajczyk Department of Ichthyobiology and Fisheries, University of Agriculture in Krakow, Poland e-mail: j.chyb@ur.krakow.pl Orexins are hypotalamic nauropeptides involved in numerous physiological processes, such as (among others) the regulation of appetite and sleep. Recent data show their role as factors influencing the secretory activity of pituitary gland. Therefore, the aim of this study was to evaluate the influence of orexin B on LH secretion from the pituitary cells of female Prussian carp (Carassius gibelio Bloch). For the experiments, the females in two different periods of reproductive cycle (resting stage of gonad development and periovulatory period) were used. After netting from pond, fish were transferred to 400 L aquaria and acclimated for 5 days. After deep anaesthesia followed by decapitation, the pituitaries were removed and transferred to an incubation medium containing antibiotic-antimycotic. After rinsing and 7-hour enzymatic dispersion of pituitaries using collagenase A, the dispersed cells were transferred to sterile 96-well microplates at amount of 50 000 cells per 250 µl of medium and subjected to 48 hours of preincubation. Then, the plates were rinsed and filled with incubation 86 medium containing orexin B at the concentrations of 10–9, 10–8 and 10–7 M or with medium containing the combination of above mentioned concentrations of orexin B and synthetic native sGnRH at the concentration of 10–8 M, each group consisted of 6 replicates. After 24-hour incubation the medium from each well was transferred to 1,5 mL Eppendorf tubes until LH determination using ELISA. In both periods of reproductive cycle orexin B stimulated spontaneous LH secretion in a dose dependent manner, being more potent in females during the resting time of gonad development, while it had no significant influence on GnRH-stimulated LH release. These results suggest that orexin B may play a positive role on Prussian carp reproduction through a direct stimulatory influence on LH secretion. Research was financed by the Ministry of Science and Higher Education of the Republic of Poland DS 3202/ KIiR. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Prenatal exposure to anti-androgen alters expression of transcription factor forkhead L2 (Foxl2) in the fetal porcine ovaries Iwona Dobosz, Katarzyna Knapczyk-Stwora Department of Endocrinology, Institute of Zoology, Jagiellonian University in Kraków, Poland e-mail: katarzyna.knapczyk@uj.edu.pl In many mammals, including pigs, formation of the primordial follicles begins during fetal development and involves the breakdown of egg nests, oocytes separation and subsequent recruitation of somatic cells (pregranulosa cells). Follicular transition from primordial to primary stage occurs when oocytes are enclosed by a single layer of cuboidal granulosa cells. It is known that transcription factor Foxl2 is a key regulator of granulosa cells differentiation. Interestingly, primordial follicle arrest occurs in Foxl2-null mice due to inhibition of ovarian granulosa cell differentiation. Previously, we have revealed that the prenatal exposure of porcine fetuses to the anti-androgen flutamide during mid- and late pregnancy resulted in delayed folliculogenesis. Thus, we hypothesized that this phenomena may, at least partially, originate from the disruption in the Foxl2 expression. In order to study the effect of anti-androgen on Foxl2 expression in the fetal porcine ovaries, pregnant gilts were injected with flutamide (50 mg/kg bw) between days 43–49, 83–89 or 101–107 of gestation. The fetal ova- ries (n = 5–12/each group) were obtained on day 50 (GD50), 90 (GD90) or 108 (GD108) of pregnancy. To assess Foxl2 expression real-time PCR and immunohistochemistry were performed. In the fetal ovaries, flutamide treatment decreased Foxl2 mRNA level in all examined groups. Foxl2 was immunolocalized exclusively in the nuclei of somatic cells surrounding ovarian egg nests in all examined fetuses. Moreover, this transcription factor was also present in the nuclei of pregranulosa and granulosa cells of primordial and primary follicles in the GD90 and GD108 groups. Concluding, diminished androgen action in porcine fetal ovaries induces downregulation of Foxl2 mRNA expression, which may affect regulation of gene expression in pregranulosa and granulosa cells and consequently lead to delay in follicle formation and its transition to the primary stage. Supported by K/DSC/002997. 87 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Does 2-hydroxyflutamide affect localization of kinases involved in non-classical testosterone pathway in prostate cancer cell line LNCaP? Ewelina Górowska-Wójtowicz1, Laura Pardyak1, Alicja Kamińska1, Anna Hejmej1, Małgorzata Kotula-Balak1, Joanna Dulińska-Litewka2, Barbara Bilińska1 Department of Endocrinology, Jagiellonian University in Kraków, Poland Chair of Medical Biochemistry, Medical College, Jagiellonian University in Kraków, Poland 1 2 e-mail: ewelina.gorowska@uj.edu.pl Testosterone has been shown to act via non-classical pathway in androgen-dependent prostate cancer cell line LNCaP. In this way, testosterone activates classical androgen receptor (AR) which triggers non receptor tyrosine kinase Src activation and stimulation of the EGF receptor (EGFR). EGFR undergoes autophosphorylation and leads to binding mitogen-activated protein kinases (MAPK: Raf, MEK and ERK1/2) to phosphotyrosines which results in CREB-dependent gene transcription. Moreover, complex between AR, Src and regulatory subunit of PI3K – p82α is required for androgen-stimulated PI3K/Akt pathway activation. The aim of this study was to assess the effect of an anti-androgen 2-hydroxyflutamide (HF) on the localization of AR and kinases involved in MAPK and PI3K/Akt pathways in LNCaP cells. The LNCaP cells were cultured in vitro and treated for 15 minutes with HF. Changes in AR and kinases 88 localization were visualized by immunocytochemistry using specific rabbit polyclonal antibodies against: AR, Akt, c-Src, Raf-1 and ERK1/2. Immunostaining for AR was localized in control and HF-treated LNCaP cells in the cytoplasm, perinuclear region and frequently as single dots in the nucleus. The positive signal of Akt was found in the cytoplasm and along the cell membrane of control cells whereas HF-treated cells exhibited exclusively diffused cytoplasmic staining. Immunopositive staining for c-Src, Raf-1 and ERK1/2 was detected in the cytoplasm of both control and HF-treated LNCaP cells. Taken together, HF administration affects localization of Akt which may suggest changes in the activation of PI3K/Akt pathway in androgen-dependent prostate cancer cell line LNCaP. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The impact of the ration supplemented with synthetic ß-carotene and silage with a pumpkin as a source of natural ß-carotene on the bioactive compounds in colostrum Gabriela Halik, Andrzej Łozicki, Agata Koziorzębska, Mateusz Makarski, Magdalena Matusiewicz Department of Feed Science and Biotechnology, Warsaw University of Life Sciences, Poland e-mail: gabrysiahalik@wp.pl Bovine colostrum has various usages, mainly is the first nutrient-rich food that can get newborn. It is easily digestible, contains easily digestible nutrients, provides the first line of defense for the content of immunoglobulin. It works such compounds as immunoglobulins, lysozym, lactoferrin makes colostrum food exceptional properties preventative and curative (Szulc 2012). Main of study was the effect of the ration of cows in the last dry period, enriched with natural and synthetic b-carotene on the bioactive composition of colostrum. The study was conducted on 40 simmental cows breed from the Experimental Institute of Animals PIB Odrzechowa. The animals were divided into four groups of 10 individuals each. The colostrum was collected from all cows up to 5 hours after calving, separated it into 50 ml tubes and then frozen at –20°C. The content of immunoglobulins, PRP, lysozym and lactoferrin was determineted. The control group(I) was feeding grass and maize silages and concentrate additive, in the II group additional 400 mg synthetic beta-carotene /day/ animal was introduced. Natural b-carotene, in the III group was introduced to the pumpkin silage (replacing it partly silage corn, so as to obtain the above and 400 mg of b-carotene to the control group), and in the IV group of 50% corn silage replaced pumpkin silage balancing b-carotene. A higher concentration of immunoglobulin found in the groups receiving silage with a pumpkin and a synthetic b-carotene. Differences statistically significant P ≤ 0.05, differences statistically highly significant P ≤ 0.01. Colostrum from group IV was characterized by the highest concentration of lactoferrin and lysozyme (P-value) differences in relation to group I. Were statistically significant differences in groups II, III, IV compared to group I. Effect of rations on the affected immunoglobulin levels in colostrum – higher levels of IgG, IgM and IgA were in the group of synthetic b-carotene, and pumpkin silage compared group I. Colostrum animals in IV group was characterized by the highest concentration of lactoferrin and lysozyme-statistically significant differences in relation to I group. 89 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of phosphodiesterase 8B ablation on mouse spermatogenesis and steroidogenesis Alicja Kamińska, Laura Pardyak, Ewelina Górowska-Wójtowicz, Małgorzata Kotula-Balak, Barbara Bilińska, Katarzyna Szczęśniak, Anna Hejmej 1 Department of Endocrinology, Institute of Zoology, Jagiellonian University in Kraków, Poland e-mail: ewelina.gorowska@uj.edu.pl The 3’–5’-cyclic adenosine monophosphate (cAMP) plays an important role in many biological processes acting as secondary messenger in intercellular signal transduction pathways. The cAMP-dependent protein kinase (PKA) signaling pathway regulates gene expression and the activity of numerous enzymes. The level of intracellular cAMP is maintained by the balance between adenylyl cyclase and phosphodiesterases. Phosphodiesterase 8B (PDE8B) is the enzyme which catalyzes hydrolysis of the cyclic phosphate bond in cAMP to generate inactive metabolite adenosine 5-monophosphate (5’AMP). The aim of the present study was to examine the role of PDE8B in the regulation of the proteins crucial for spermatogenesis (androgen receptor (AR) and N-cadherin) and steroidogenesis (StAR, 3b-hydroxysteroid dehydrogenase and aromatase) in mouse testis. The study was performed on testicular tissue isolated from adult Pde8b knock-out mice (KO) and wild-type C57BL/6 mice. Morphological analysis was carried out by hematoxylin-eosin staining. Locali- 90 zation and expression of the proteins were analyzed using immunohistochemistry and Western blot, respectively. In the testis of KO mice morphological alterations of the seminiferous epithelium were observed. Immunohistochemical analysis showed delocalization and decreased staining intensity of N-cadherin and AR in KO testis when compared to the controls. Decrease of AR, N-cadherin, StAR and aromatase protein, and increase of 3bHSD protein expression were demonstrated in the testis of KO mice (*P < 0.05) compared to wild-type mice by WB analysis. In summary, disturbances in the structure of the seminiferous tubules, altered localization and expression of studied proteins observed in KO indicate that PDE8B is an important regulator of cAMP pool involved in the control of both spermatogenesis and steroidogenesis. Supported by K/ZDS/005402. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Comparison of antioxidant status in farmed raccoon dog from Poland and Russia Tatiana N. Ilyina1, Irina V. Baishnikova1, Viktor A. Ilyukha1, Svetlana N. Sergina1, Stanisław Łapiński2 1 Institute of Biology, Karelian Research Centre, Russian Academy of Sciences, Petrozavodsk, Russia 2 Department of Swine and Small Animals Breeding, Institute of Animal Sciences, University of Agriculture in Krakow, Poland e-mail: ilyina@bio.krc.karelia.ru Changes in natural photoperiod trigger modifications in many physiological, morphological, and behavioral processes that are believed to promote survival in seasonally breeding animals. Seasonality results in metabolic adjustments that include changes in the antioxidant defense system that protects organisms against the excess of reactive oxygen species and this plays an important role in adaptation to environmental factors. The aim of the investigation was to conduct a comparative study of various indicators of the antioxidant system activity in farmed raccoon dog (Nyctereutes procyonoides Gray) from Poland and Russia in autumn period. Tissue samples (liver, kidney, heart, lung, spleen, skeletal muscle) were collected in the late autumn when animals show willingness to winter conditions and examined for indicators of the antioxidant system – vitamin A and E content, reduced glutathione (GSH) level, activities of super- oxide dismutase (SOD) and catalase. The data suggest differences in some indicators between animals from the two regions. The largest differences between values were observed in the liver and skeletal muscles. Liver SOD activity, GSH content and vitamin levels were significantly higher in animals from Russia than in raccoon dogs from Poland. Also skeletal muscle of raccoon dog from the colder region had higher enzyme activities but lower α-tocopherol level than animals from the warmer region. However, no differences in the values between animals from different localities were detected in heart that demonstrated high stability of antioxidant defense in this tissue. Results suggest that raccoon dogs from higher latitudes possess stronger antioxidant capacity than animals from lower latitudes. These differences appear to be related to the availability of energy and the relative severity of climatic conditions in Russian locality. 91 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Agonist and antagonist of 5-HT2B/2C receptor and ghrelin activate intracellular signal cascades responsible for melatonin biosynthesis in the ovine pineal gland Katarzyna Kirsz, Małgorzata Szczęsna, Dorota A. Zięba Department of Animal Biotechnology, University of Agriculture in Krakow, Poland e-mail: k.kirsz.ur@gmail.com Recent research has indicated a participation of ghrelin (GHRL) and serotonin receptors type 2 in the regulation of melatonin (MEL) biosynthesis and secretion from the ovine pineal gland (PG). Herein, we examined the role of GHRL, mCPP (a mixed agonist of the 5-HT2B/2C receptor) and RS 102221 (a highly selective antagonist of the 5-HT2C receptor) in the activation of inositol 1,4,5-triphosphate (IP3), diacylglycerol (DAG) and protein kinase C (PKC), leading to release of MEL from the ovine PG explants. Pineal glands were collected after sunset from 8 ewes during short days (November). The PG were transected sagittally into strips (~ 30 mg) with each equilibrated in 1.0 ml of DMEM for 60 min, followed by a 4-h incubation in medium alone (Control) or in medium containing: GHRL (100 ng/ml); GHRL (100 ng/ml) + m-CPP (10 µM); RS 102221 (10 µM) +m-CPP (10 µM). The PG explants were harvested every 60-min, frozen in liquid nitrogen and stored at –80°C until ELISA for IP3, DAG and PKC determinations. Samples of medium were stored at –20°C until RIA for 92 MEL. There was no effect of experimental factors on IP3 concentrations in the culture of the PG explants. However, treatment with GHRL decreased (P < 0.05) DAG concentrations as compared with controls and GHRL + mCPP group. The RS 102221 + mCPP combination reduced (P < 0.05) DAG and PKC concentrations in comparison to all experimental groups. Exogenous GHRL and RS 102221 + mCPP inhibited (P < 0.05) the secretion of MEL as compared to controls, whereas treatment with GHRL + mCPP increased (P < 0.01) the concentration of MEL in relation to all experimental groups. These results indicate that GHRL and 5-HT2 receptors are involved in the regulation of MEL biosynthesis and secretion through activation of DAG/PKC intracellular signal cascade. This work was supported by grant from the National Science Center (NCN 2012/05/B/NZ4/02408) and the Ministry of Science and Higher Education of Poland (BM4241/KBZ/2015). International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 T FOXO3 genetic variability in selected Polish sheep breeds Urszula Kaczor1, Magdalena Luber1, Mirosław Kucharski1, Andrzej Kaczor2 1 Department of Animal Biotechnology, University of Agriculture in Krakow, Poland 2 Department of Technology, Ecology and Economics of Animal Production, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: rzkaczor@cyf-kr.edu.pl FOXO proteins are a subgroup of the Forkhead family of transcription factors. This family is characterized by a conserved DNA-binding domain (the ‘Forkhead box’, or FOX) and comprises more than 100 members in humans, classified from FOXA to FOXR on the basis of sequence similarity. In invertebrates, there is only one FOXO gene, termed daf-16 in the worm and dFOXO in the fly. Intriguingly, FoxO transcription factors extend longevity in invertebrates. In mammals, there are four FOXO genes, FOXO1, 3, 4, and 6. FOXO transcription factors are at the interface of crucial cellular processes, orchestrating programs of gene expression that regulate apoptosis, cell-cycle progression, oxidative stress resistance, and may also extend mammalian lifespan. They also regulate energy metabolism and development of a number of tissues. Recent studies also provide some evidence for an association between ageing, human longevity, and variation in the FOXO3 gene. The genetic variability in FOXO3 may potentially affect sheep lifespan either directly or indirectly by impacting on factors that lead to reduced productivity and increased likelihood of culling. The aim of the study was to identify polymorphisms in exon 2 of the FOXO3 gene in 90 domestic sheep of three breeds: Polish Mountains sheep, Suffolk meat sheep, and prolific Olkuska sheep. The test gene consists of 4 exons wherein exon 2 encodes coding sequence domains of FH (well folded Forkhead DNA-binding domain) and conserved regions CR2 or CR3, including DNA binding sites of other genes. This region is highly conservative in many species. The method of PCR-MSSCP was used and the presence of two patterns of MSCP emerging from different turnouts was revealed. The sequencing of the PCR product patterns varying in MSSCP showed the presence of SNPs which change the amino acid sequence of synthesized proteins of p. 285 T > I (g. 845A > C) and p. 288 E > D (g. 825 C > G) (NCBI: NM_001267889.1). 93 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Identification of SNP c.*1116A > G and c.2218G > A in the SCD gene of sheep breeds raised in Poland Mirosław Kucharski1, Urszula Kaczor1, Katarzyna Piórkowska2, Karolina Pastuszak1 Department of Animal Biotechnology, University of Agriculture in Krakow, Poland Department of Genomics and Molecular Biology, National Research Institute of Animal Production, Balice near Krakow, Poland 1 2 e-mail: mirkuch@op.pl Stearoyl-CoA desaturase encoded by the SCD gene is an oxidoreductase class enzyme that catalyzes the introduction of the first double bond in fatty acids. Due to its role, stearoyl-CoA desaturase influences the fatty acid profile of adipose tissue and determines the quality of animal derived products, i.e. meat and milk. SCD protein mainly determines the ratio of saturated (SFA) to unsaturated fatty acids (UFA). The aim of the study was to analyse genetic variation in the stearoyl-CoA desaturase gene. Biological material was collected from 150 sheep aged 2–3 years, which belonged to five breeds: Ile de France, Kołudzka, Olkuska, Polish Mountain Sheep, and Suffolk. Genomic DNA was isolated from peripheral blood collected at one time from each animal. High resolution melting analysis (HRMA) as a method of screening for polymorphic sites was used 94 to study genetic variation in 11 fragments containing regulatory, coding sequences and splicing sites of the SCD gene. Sequencing of selected samples showed the presence of two mutations, c.*1116A > G and c.2218G > A, in the 3’-UTR region. RFLP-PCR reactions using RsaI and BccI enzymes, respectively, were also designed. For the c.*1116A > G mutation, alleles A (0.42) and G (0.58) and genotypes GG (0.33), GA (0.49) and AA (0.18) were identified. Analysis of the occurrence of c.2218G > A substitution revealed that only one animal (of the Suffolk breed) has the GA genotype, and no homozygote with the AA mutation was found in any of the breeds. The results obtained will serve to show the possible effect of the c.*1116A > G mutation in the region of the stearoyl-CoA desaturase gene on sheep meat quality. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Identification of SNP c.391 – 66C > T and c.529 + 27G > C in the PPARγ gene of sheep breeds raised in Poland Mirosław Kucharski1, Urszula Kaczor1, Katarzyna Piórkowska2, Katarzyna Zemsta1 Department of Animal Biotechnology, University of Agriculture in Krakow, Poland Department of Genomics and Molecular Biology, National Research Institute of Animal Production, Balice near Krakow, Poland 1 2 e-mail: mirkuch@op.pl Peroxisome proliferator-activated receptors (PPAR) are ligand-activated transcription factors that play a significant role in several biological pathways associated with lipid, protein and carbohydrate metabolism. PPARγ-activated receptors play a key role in adipogenesis and lipogenesis. In addition, they contribute to oxidation of long-chain fatty acids and are responsible for enhancing gluconeogenesis and increasing insulin sensitivity. The PPARγ protein coding gene is responsible for lipid metabolism, thus affecting the content of individual fatty acids in depot fat and in the intramuscular fat of muscle tissue. Biological material was collected from 150 sheep aged 2–3 years, which belonged to five breeds: Ile de France, Kołudzka, Olkuska, Polish Mountain Sheep, and Suffolk. Genomic DNA was isolated from peripheral blood. High resolution melting (HRM) analysis was used to investigate genetic variation in 8 fragments containing regulatory, coding sequences and splicing sites of the PPARγ gene. Sequencing of selected samples showed the presence of c.391 – 66C > T and c.529 + 27G > C mutations in the region of introns 4 and 5. RFLP-PCR reactions using the BccI and HpyCH4III. enzymes, respectively, were designed to identify mutations in the entire population. In the analysed population, diversified in terms of breeds, identical allele and genotype distributions were obtained for the c.391 – 66C > T and c.529 + 27G > C mutations: alleles C/G = 0.10 and T/C = 0.90 and genotypes CC/GG = 0.01, CT/GC = 0.18 and TT/ CC = 0.81. In the case of both SNPs, the high frequencies of the alleles with the mutation may be indicative of significant differences in the genetic structure of the analysed breeds compared to the Iranian breeds of sheep in which these mutations were identified for the first time. 95 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Analysis of the structure of sperm chromatinin breeding foxes Marta Kuchta-Gładysz1, Katarzyna Andraszek2, Piotr Niedbała2, Agnieszka Otwinowska-Mindur3, Anna Grzesiakowska1, Olga Szeleszczuk1 Department of Animal Anatomy, University of Agriculture in Krakow, Poland Department of Swine and Small Animal Husbandry, University of Agriculture in Krakow, Poland 3 Department of Genetics and Animal Breeding, University of Agriculture in Krakow, Poland 1 2 e-mail: m.kuchta-gladysz@ur.krakow.pl While testing semen of breeding animals, steps are taken towards possibly most accurate evaluation of sperms’ ability to fertilization. The most recent reports indicate the fact that the structure of sperms’ chromatin and DNA integrity significantly influence sperms’ functions. The aim of the research was to determine condensation level in the structure of chromatin of fresh semen as well as the damage degree in DNA chain of the chilled semen of tested breeding foxes. The semen for analysis was collected by means of a manual method while insemination procedure used on the farm. 38 foxes including 15 Alopex lagopus and 23 Vulpes vulpes were used in the experiment. Sperms of one-year breeding foxes were the subject of survey. Directly after collecting, macroscopic and microscopic evaluation of the semen with concentration of 15 × 106 sperms/L mL of semen was performed. Estimation of chromatin’s structure was conducted by means of three techniques of staining: aniline blue (AB, Franken et al. 1999), chromomycin (CMA3, Lolis et al. 1996) and acridine orange (AO, Tejda et al. 1984). To evaluate differences in DNA integrity of the fresh sperms (0h) and the chilled ones (4º C for 24h / 48h / 72h), the semen was diluted to the concen- 96 tration of 10 × 106/5µl and the comet test was used (SCGE, Singh et al. 1988). The sperms were evaluated by means of the fluorescent microscope Olympus BX 50, using the lens Olympus UplanApo 100x/1.35/Oil Iris/∞/0.17. 500 sperms stained with each method were evaluated for each animal. The difference between species were investigated by the one-way analysis of variance with repeated measurements, using SAS. The Tukey Kramer test was used for multiple comparison. The average percentage of head DNA was changing during chilled: in the case of blue foxes from 99.12% (SE = 0.038%) to 97.83% (0.062%) and in Vulpes vulpes decreased from 99.37% (SE = 0.028%) to 98.05% (SE = 0.045%). When the head DNA was analyzed, the highly significant difference (P < 0.01) between species was observed. Similarly, the head DNA differed highly significantly (P < 0.01) in chilled semen (0h / 24h / 48h / 72h), within species. Sperms with proper histones retention were mostly observed in the result of staining by AB, CMA3 and AO. The average share of sperms with increased, improper content of histones was on the similar levels regardless the staining method, both in Vulpes vulpes (0–4%) and Alopex lagopus (0–3%). International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of butylated hydroxytoluene on bioavailability of dietary nutritional compounds Magdalena Mika, Agnieszka Wikiera, Anna Antończyk, Anna Starzyńska-Janiszewska Department of Food Biotechnology, University of Agriculture in Krakow, Poland e-mail: m.mika@ur.krakow.pl The addition of butylated hydroxytoluene (BHT) to the diet in animal studies may influence the bioavailability of nutritional compounds. A diet applied in nutritional studies is usually prepared once at the beginning of an experiment and thus it is stabilized with preservatives. Antioxidants are added at small amounts of 0.001–0.01%. The results of the in-vitro study showed that the addition of BHT at a dose of 0.01% to the diet containing butter and wheat roll (1:4) resulted in the increase of fat, carbohydrates and protein bioavailability. In order to estimate the bioavailability of dietary compounds, the model product was subjected to the in-vitro digestion and next the hydrolysate was analysed for the intestinal absorption (using Caco-2 cell monolayers). The sample with BHT addition was characterised by the higher bioavailability of protein, glucose, cholesterol and triacylglycerols as compared to the control one. BHT was proven to inhibit the activity of digestive enzymes when applied at 10 times higher concentration than the one used in our study. It is also established that the peroxidative attack, initiated by Fe2+-ascorbate, resulted in the reduction of sodium-dependent glucose transport and lowered the bioavailability of lipids. However, in the case of our study no differences in the concentration of lipid peroxidation products were observed between the sample containing BHT and the control. Thus, further studies concerning the mechanism of BHT effect on the absorption of nutrients from the digestive tract are needed. 97 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The role of light and exogenous melatonin in the regulation of prolactin secretion in sheep Edyta Molik1, Michał Błasiak1, Tomasz Misztal2, Katarzyna Romanowicz2, Dorota A. Zięba1 Department of Animal Biotechnology, University of Agriculture in Krakow, Poland Department of Endocrinology, The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, Jablonna near Warsaw, Poland 1 2 e-mail: rzmolik@cyf-kr.edu.pl In seasonal breeding animals, the basic environmental stimulus regulating sexual activity of the body is changing the length of day light. The aim of the study was to determine the effect of exogenous melatonin on prolactin secretion in sheep during pregnancy. The material of experiment was 60 ewes (Polish Longwool Sheep). Animals were divided into 3 groups (Group 1 : n = 20, Group 2 : n = 20, Group 3 : n = 20). Fertilization for the first group was carried out on September 15, lambing were held on 15 February. For Group 2 and 3 were performed on 15th January. The measurement of concentration of melatonin (MLT) and prolactin (PRL) carried out by RIA method. The first blood sampling was performed 6 weeks before lambing in all groups of sheep, then mothers Group 3 implanted subcutaneous implants of melatonin (Ceva Animal – Melovine 18 mg). The second blood sampling carried out in 57 day of lactation. Blood samples were collected each time, every 10 minutes, from the 98 time of the sunset through a catheter inserted into the jugular vein. 6 weeks before lambing highest concentration of MLT occurred in animals Group 1 (January) (148.15 ± 12.4 pg/ml), while the lowest in sheep Group 2 (May) (68.0 ± 7.6 pg/ml). The lowest concentration of prolactin has been found in animals in Group 1 (12.11 ± 2.6 ng/ml), and the highest in sheep Group 3 (322.0 ± 18.1 ng/ml). The studies from 57 day of lactation have shown that the highest concentration of MLT occurred in sheep Group 3 (106.4 ± 10.7 pg/ml), and the lowest in Group 2 (22.44 ± 8.75 pg/ml). By analyzing changes in the levels of prolactin in the test group of sheep in a 57 day lactation, it was shown that the highest concentration of PRL occurred in animals from Group 1 (110.55 ± 5.5 ng/ml), while the lowest in sheep Group 3 (17.17 ± 2.07 ng/ ml). The study showed that the presence of exogenous melatonin during pregnancy reduced the secretion of prolactin in 57 day of lactation. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Cryopreservation of bull semen produces an oxidative damage of sperm proteins Agnieszka Mostek, Mariola A. Dietrich, Błażej Westfalewicz, G. Hołownia, Andrzej Ciereszko Institute of Animal Reproduction and Food Research of Polish Academy of Science, Olsztyn, Poland e-mail: a.mostek@pan.olsztyn.pl Artificial insemination is biotechnological advancement that has made a significant contribution to the genetic improvement, particularly in bulls breeding. The technique of artificial insemination could not be conducted on a large scale without cryopreservation of sperm. However, cryopreservation reduces the functional and structural integrity of bull spermatozoa, and is associated with reactive oxygen species (ROS) production. Oxidative stress during freezing–thawing process of mammalian semen can cause functional and structural damage to spermatozoa that impair sperm motility, membrane integrity, and fertilizing potential. The aim of this study was to investigate the oxidative modifications of bull spermatozoa proteins triggered by cryopreservation process. Western blotting, in conjunction with 2D PAGE (2D-Oxyblot) and MALDI TOF spectrometry, was employed to identify and quantify the specifically-carbonylated spermatozoa proteins. We identified 13 protein spots representing 8 proteins showing significant (p < 0.05) differences in protein oxidation in response to cryopreservation. These proteins include ropporin, glutathione-S-transferase, phosphatidylethanolamine binding protein, actin binding protein, actin related protein, calicin, triosephosphate isomerase and uncharacterized protein C15orf26. Many among the identified proteins are associated with actin what suggests oxidative damage of cytoskeleton during cryopreservation process. In conclusion, we have demonstrated for the first time that cryopreservation of bull semen produces an oxidative damage of specific sperm proteins. Further research is needed to explore the mechanisms of the particular protein oxidation changes and to establish the relationship between those changes and sperm quality. 99 Kriokonserwacja nasienia buhaja powoduje oksydacyjne uszkodzenia białek plemnikowych Agnieszka Mostek, Mariola A. Dietrich, Błażej Westfalewicz, G. Hołownia, Andrzej Ciereszko Instytut Rozrodu Zwierząt i Badań Żywności Polskiej Akademii Nauk w Olsztynie e-mail: a.mostek@pan.olsztyn.pl Sztuczne zapłodnienie jest powszechnie stosowaną biotechniką w rozrodzie zwierząt gospodarskich, która przyczyniła się do genetycznego udoskonalenia populacji zwierząt gospodarskich, w szczególności bydła. Technika sztucznego zapłodnienia nie mogłaby być prowadzona na szeroką skalę bez kriokonserwacji nasienia. Jednakże kriokonserwacja zmniejsza funkcjonalną i strukturalną integralność plemników byka, co wiąże się z wytwarzaniem reaktywnych form tlenu (ROS). Stres oksydacyjny podczas procesu zamrażania i rozmrażania nasienia ssaków może powodować uszkodzenia funkcjonalne i strukturalne plemników, które upośledzają ich ruchliwość, integralność błony komórkowej oraz zdolność do zapłodnienia. Celem niniejszej pracy było zbadanie zmian poziomu utlenienia białek plemnikowych, wywołanych przez proces kriokonserwacji. Do identyfikacji i określenia specyficznych zmian karbonylacji białek plemników zastosowano metodę western blotting, w po- 100 łączeniu z elektroforezą żelową 2D PAGE (Oxyblot) oraz spektrometrią mas MALDI TOF. Zidentyfikowano 13 plam białkowych reprezentujących 8 białek wykazujących istotne (p < 0,05) różnice w poziomie utlenienia białek w odpowiedzi na kriokonserwację nasienia. Do białek tych należą: roporyna, transferaza glutationowa, białko wiążące fosfatydyloetanoloaminę, białko wiążące aktynę, białko związane z aktyną, calicyna, izomeraza triozofosforanowa, niescharakteryzowane białko C15orf26. Wiele spośród zidentyfikowanych białek jest powiązanych z aktyną, co sugeruje występowanie oksydacyjnych uszkodzeń cytoszkieletu w procesie kriokonserwacji. Podsumowując, po raz pierwszy wykazano, że kriokonserwacja plemników byka powoduje oksydacyjne uszkodzenia poszczególnych białek plemników. Konieczne są dalsze badania ukierunkowane na zbadanie mechanizmów poszczególnych zmian utleniania białek oraz relacji pomiędzy tymi zmianami a jakością nasienia. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The pre-established reserve of porcine preantral follicles – advanced method of enzymatic isolation and identification Maria Motal, Kamil Wartalski, Małgorzata Duda Department of Endocrinology, Institute of Zoology, Jagiellonian University in Kraków, Poland e-mail: kamil.wartalski@doctoral.uj.edu.pl Proper follicular development is necessary for the emergence of functional oocytes. Growth and development of the follicle from the primary to the mature Graafian follicle stage requires the presence of multiple regulatory factors. Knowledge of the mechanisms allowing for the proper development of preantral follicles under physiological conditions is important because on its basis in vitro follicular maturation can be performed. So far, the greatest difficulty in the studies on in vitro preantral follicular development was low isolation efficiency and low survival rate of cultured structures. Atresia of follicles was caused by their improper isolation from ovarian cortex, resulting in their damage and incapability of development and survival. The purpose of this study was to develop an efficient and effective method of preantral follicles isolation from the ovarian tissue of sexually immature pigs. After isolation of the cortical part of the ovary obtained from slaughterhouses tissue was cut into small pieces. Slices were placed in a variety of enzymes, to determine both safe and effective mixture. After that, histochemical and immunohistochemical staining was performed to determine the morphology and ultrastructure of isolated follicles. Results of this study confirmed the effectiveness of the enzymatic isolation of preantral follicles of large mammals. The most effective isolation of the preantal follicles was achieved with liberase. Microscopic observations has proven that this enzyme allowed for obtaining high amounts of integral follicles. Because of the preservation of the correct morphology and ultrastructure enzymatically isolated follicles are likely to be successfully used for further in vitro development. Receiving follicles under extracorporeal conditions requires further research on the factors involved in the process of follicular development. Supported by DEC-2013/09/B/NZ9/00226 from National Science Center Poland. 101 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Usefullness of disected sheep heads in paranasal sinuses endoscopy for physicians Klaudia Nalik1, Kinga Fic1, Patryk Hartwich2, Paweł Stręk2, Michał Nowakowski1, 3 1 Center for Experimental and Innovative Medicine, University Center for Veterinary Medicine UJ-UR, Krakow, Poland 2 Chair and Department of Otolaryngology, UJ CM, Krakow, Poland 3 Department of Medical Didactics, UJ CM, Krakow, Poland e-mail: k.nalik@ur.krakow.pl Endoscopy is one of the least invasive methods of diagnosing and treatment of nose and sinus diseases. Proper personnel training continues to be the biggest obstacle for its widespread use. Performing surgeries within a narrow visual field, propriosensoric dissociation and lack of 3D vision, seem to be the biggest obstacles in this area. Training during actual surgeries is a controversial topic, so searching for an adequate animal model has a high priority. Purpose: Evaluation of using sheep heads as a training method for: (1) increasing the ability to use equipment, (2) increasing manual dexterity, (3) planning procedures. A group of 48 laryngologists participated in training of the use of medical equipment using prepared sheep heads as a model. Groups of 12 people were divided into pairs and trained simultaneously, supervised by two laryngologists and two technicians. Effects of training were then evaluated by the supervisors. Sheep heads were prepared using R. Mla- 102 dina protocol. Karl Storz equipment was used for training. All of the personnel finished the training. 56% of participants rated the animal model as adequate and 21% as good or very good representation of working with humans. 52% of participants stated that the sheep material allows to adequately train planning and performing procedures on humans, 43% rated it as good or very good. Training of endoscopy equipment usage (82% participants rated it as good or very good) and increasing manual dexterity (82% participants rated it as good or very good, 18% rated it as adequate) showed best results. According to supervisors, everyone from the study group increased their abilities significantly. Before training, manual dexterity of the participants was rated as poor for 52% of the group and 48% as good or adequate. After training 28% were rated as poor and 72% as good or adequate. Prepared sheep heads are adequate training model for physicians. Training methodology seems to have room for optimization. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Preliminary research of chosen biochemical compounds’ level in the semen of breeding chinchillas (Chinchilla lanigera) Piotr Niedbała1, Gabriela Lipień1, Olga Szeleszczuk2, Marta Kuchta-Gładysz2, Stanisław Łapiński1, Iwona Guja1 1 Department of Swine and Small Animal Husbandry, University of Agriculture in Krakow, Poland 2 Department of Animal Anatomy, University of Agriculture in Krakow, Poland e-mail: p.niedbala@ur.krakow.pl The aim of the surveys was to determine fructose and citric acid levels in breeding chinchilla males semen plasma. The tests were performed in 40 ejaculates collected by means of electroejaculation method from young chinchilla males 9–10-month-old (n = 20) and animals at the age from 2 to 8 years (n = 20). The semen was collected in two periods: summer–autumn (n = 20) and winter–spring (n = 20). The level of fructose was determined using Mann’s method and the level of citric acid by Speck’s method with Mann’s modification. Preliminary surveys showed that the content of fructose in semen plasma stood at the average level of 241 in the range from 68 to 538 mg/100 ml. Its slightly higher content was stated in the ejaculates of 9–10-month-old males soon after obtaining sexual maturity. At the time, it accounted for the average level of 270 mg/100 ml. In the semen plasma of multiannual males, its amount was 211 mg/100 ml. Small differences between seasons were also observed. Slightly higher rate of mean fructose amounting to 248 mg/100 ml was stated in the winter–spring period than in the summer–autumn one (233 mg/100 ml). The average level of citric acid which testi- fies to androgenic activity of testicles amounted to 0.5 mg/100 ml of plasma in the range from 0.081 to 1.16 mg/100 ml. The level of this compound in both groups was approximate and respectively amounted to 0.493 mg/100 ml in younger animals and 0.507 mg/100 ml in multiannual ones. However, bigger differences were observed between seasons. In the summer–autumn one, the level of citric acid amounted to 0.471 mg/100 ml and in the winter–spring one it was 0.533 mg/100 ml. Preliminary surveys showed that the age of animals as well as the season do not influence significantly the change of fructose and citric acid levels in plasma of breeding chinchillas’ semen. Only the significant correlation (r = 0.731) between fructose and citric acid levels in the semen plasma of 9–10-month-old males in the winter–spring period was observed. Supported by the Polish Ministry of Science and Higher Education/National Science Centre under project No NN311297235 and University of Agriculture Research Funding No DS 3266/DSSAB/2015. 103 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Analysis of KM1 allele frequency in intron 13 of the KIT gene in solid-coloured Hucul horses with white markings Marta Pasternak Department of Animal Genetic Resources Conservation, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: marta.pasternak@izoo.krakow.pl Mutations in the KIT gene, which affect hair and coat pigmentation have been observed in many studies with humans and different species of animals such as mice, pigs or horses. One such change is substitution in intron 13 of the KIT gene in horses with tobiano pattern. The mutation creates a cutting site for the restriction enzyme MspI. Earlier research showed, however, that the presence of KM1 allele (with an additional cutting site) cannot be considered the direct cause of piebaldism because it is also found in some horses of solid colour. The aim of this study was to analyse KM1 allele frequency in solid-coloured Hucul horses with white markings, for the purpose of determining the relationship between this mutation and the occurrence of undesirable markings on legs and head. Blood samples collected from the horses were analysed by PCR-RFLP. Genotype frequency in the 104 experimental group (52 solid-coloured horses with white markings) was 0.39 for KM0 homozygotes, 0.48 for KM0/KM1 heterozygotes and 0.13 for KM1 homozygotes. In the control group (61 solid-coloured horses without white markings), the frequency was 0.53 for KM0 homozygotes, 0.39 for KM0/KM1 heterozygotes and 0.08 for KM1 homozygotes. Allele frequencies were 0.63 for KM0 and 0.37 for KM1 in the experimental group and 0.74 for KM0 and 0.26 for KM1 in the control group. Based on the results obtained, no clear relationship was found between KM1 allele and the occurrence of white markings in the studied horse group. Comparison of the results with phenotypic evaluation also revealed no correlation between genotype variant and the size and extent of markings. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 In vitro LH secretion by female common carp (Cyprinus carpio L.) pituitary cells incubated with popular herbicide Roundup Magdalena Socha, Jarosław Chyb, Mirosława Sokołowska-Mikołajczyk, Ewa Drąg-Kozak, Włodzimierz Popek Department of Ichthyobiology and Fisheries, University of Agriculture in Krakow, Poland e-mail: m.socha@ur.krakow.pl The aim of the study was to determine the direct effect of Roundup on LH secretion by pituitary cells collected from common carp (Cyprinus carpio L.). In Poland, Roundup was sold with the information connected with its biodegradation, but it was shown that it did not degrade in natural environment. Roundup was detected in the tissue of animals and humans, moreover it was also shown that this herbicide has hormonal activity. Roundup might be used to control aquatic vegetation in ponds, lakes so it was interesting to investigate its effect on fish reproduction. Gonadotrophs were obtained from 3 sexually mature female of Common carp (mean body weigh about 4.24 kg) at the time of natural spawning (June). Gonad maturity was specified as a percentage of body weight (gonadosomatic index GSI) and was about 12.87%. Collected pituitary glands were chopped into small pieces and exposed to medium containing collagenase A. The dispersed pituitary cells were incubated with different concentrations of Roundup (10, 100 and 1000 ng mL-1 medium) or/and salmon gonadotropin-releasing hormone analogue (sGnRHa) at the concentration of 10-7 M. The luteinizing hormone (LH) levels in the cultured medium were measured after 24 hours of incubation by ELISA method. After 24 hours of incubation in the control culture the LH level was 24,303 ng mL-1 medium. Statistically important stimulation (33.99 ng mL-1 medium) was observed in group incubated with sGnRHa as compared to the control. Roundup given at the highest concentration caused statistically significant increase in LH secretion (44.88 ng mL-1 medium) after 24 h of incubation in comparison to control group as well as to the lowest tested concentration of Roundup. Obtained results show the possible impact of Roundup on (female) common carp reproduction at the level of pituitary, by direct stimulating action on LH secretion, what might suggest more careful using of this popular herbicide specially in the area close to fish farm. 105 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Application of time-lapse monitoring of embryo development (TLMED) in different mouse strains Grzegorz Sokołowski1, Ricardo Faundez2, Zuzanna Sandowska-Markiewicz1, Marta Gajewska1, Anna Niwińska2, Anita Tyl-Bielicka1, Marek Woszczyński1, Katarzyna Kermen-Germaniuk1, Sebastian Dąbrowski2, Monika Petrajtis-Golobow2, Jerzy Ostrowski1 1 The Department of Genetics, Maria Skłodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland 2 The Department of Large Animal Diseases with Clinic, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Center for Biomedical Research (CBB-SGGW), Warsaw, Poland e-mail: gsokolowski@coi.waw.pl Time Lapse Monitoring of Embryo Development – TLMED – is the imaging system to observation the exact sequence of consecutive events in real-time. Monitoring of embryo development is the new method which permit an appropriate embryo selection by the identification of the embryo showing the highest developmental potential. The purpose of this study was to check the developmental potential of embryos from different mouse strains, using TLMED technique. Animals were 3 (8–10 weeks aged) female mice from every A.CA/W, BALB/cW, C57BL/6W, C3H/W, C3Hwad/W, CBAmut/W, superovulated (eCG-5 IU, hCG-5 IU with 42–48-hour interval), mated with the male mice form the same strain. After 24 hours, animals were euthanized, necropsy was performed and embryos were flushed out from the oviducts as routine breeding procedures. Only 1-, 2-, 3- cell cleavages with normal morphology were obtained (abnormal or unfertilized were discarded). Embryos were cultured in the CSC medium containing 10% SSS (Irvine sci, USA) for 96–120 hours in three gas atmosphere (6% 106 CO2, 5% O2, 89% N2) incubator. The morphometrical imaging and assessment were provided using Primo Vision Time-Lapse Embryo Monitoring System (Vitrolife, Sweden). From C3H/W and C3Hwad/W it was obtained respectively 9/16 and 11/16 blastocysts. C57BL/6W and CBAmut/W are characterised by lower fraction of embryos with high developmental potential – 3/8 and 6/16 blastocysts obtained. After 96-hour culture, most of BALB/cW and A.CA/W embryos were dead or stopped the development at early stages. Re-analysis with strong pre-selection of embryos (only 2-, 3-cell embryos were cultured with Primo Vision monitoring) allowed to obtain later stages – 3/60 blastocysts form A.CA and 2/40 form BALB/cW). Embryos form different mouse strains show variable developmental potential and timing. Time-lapse analyze of embryo development may be crucial method for selection of embryos with high developmental and implantation potential to increase the efficiency of cryo-banking and embryo transfer. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The role of cumulus cells in in vitro maturation and in vitro fertilization of oocytes of domestic cat Natalia Sowińska, Anna Filipczyk, Katarzyna Frankowska, Klaudia Nalik, Kinga Fic Center of Experimental and Innovative Medicine, University Center of Veterinary Medicine UJ-UR, Krakow, Poland e-mail: kinga.fic07@gmail.com Cumulus cells (CC) are important for oocytes growth, maturation and fertilization. They supply the oocytes with nutrients, but during cryopreservation they decrease the permeation of the cryoprotectant into the oocytes, affecting the results. Studies performed on cat oocytes showed positive influence of some co-culture systems on resumption of meiosis and fertilization. The aim of the present study was to evaluate the effect of the co-culture of denuded oocytes with CC or COC on the results of in vitro maturation (IVM) (experiment 1) and in vitro fertilization (IVF) (experiment 2). Immature oocytes were collected from ovaries of domestic cat. In experiment 1, oocytes were in vitro matured for 24 h in four groups: (i) DO, (ii) DO co-cultures with CC, (iii) DO co-cultured with COC, (iv) COC as a control group and then denuded mechanically after incubation in 0.2% hyaluronidase. In experiment 2, COC were IVM for 24 h, then oocytes were divided into four groups analogous to that in exp. 1 and IVF. Presumptive zygotes were cultured up to 7 days. In the end of each experiment oocytes/embryos were stained in Hoechst 33342 (2 µg/ml solution) and observed under the epifluorescence microscope. In experiment 1 of maturation rate of oocytes decreased significantly in all experimental groups, when compared to the control group. Maturation rate amounted 21% (5/24), 14% (4/28), 33% (8/24) and 50% (18/36) (p < 0.05), in a group (i), (ii), (iii) and (iv), respectively. In experiment 2, embryos up to morula stage developed in all experimental groups. DO and oocytes cultured with COC during fertilization showed lower cleavage rate – 36% (9/25) and 25% (3/12) than those co-cultured with loose CC and from the control group 43% (12/28) and 42% (16/38), respectively. Results of this study show that cumulus cells connected with oocyte into an cumulus oocyte complex are irreplaceable in the process of maturation of domestic cat oocyte, but addition of loose CC might be useful in the process of IVF. 107 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Genetic variability in equine GDF9 in Arab and Thoroughbred mares Monika Stefaniuk-Szmukier1, Katarzyna Ropka-Molik2, Katarzyna Piórkowska2, Jarosław Łuszczyński1, Magdalena Pieszka1, Tomasz Szmotała2, Bogusława Długosz1, Romana Augustyn1, Weronika Pisarczyk1, Zenon Podstawski1 Department of Horse Breeding, University of Agriculture in Krakow, Poland Department of Animal Genomics and Molecular Biology, National Research Institute of Animal Production, Balice near Krakow, Poland 1 2 e-mail: zpodstawski@ar.krakow.pl In horses, multiple ovulation resulting implantation of multiple embryos are adverse. It has been postulated that multiple ovulation (MO) may have genetic background. Despite the low heritability rate there is few evidence that this trait is incorporated with sire lines. Numerous mutations has been found across species associated with premature ovarian insufficiency, twinning’s, infertility or ovarian hiperstymulation syndrome. Two major: genes: bone morphogenetic protein 15 (BMP15) and growth and differentiation factor 9 (GDF9) are recognized as playing a crucial role in folliculogenesis and controlling the ovulation rate. Thus the aim of presented study is structural analysis of GDF9 in horses. Besides we perform variation screening of investigated gene in population of TB vs. Arab horse breeds as Thoroughbred which are recognized as having high rate of multiple pregnancy (20 %) whilst Arabs are recognized with low estimation of this trait. DNA samples from five mares with confirmed spontaneous, repetitive multiple ovulation (at least 5 108 seasons of repetitive multiple ovulation in cycles under control of ultrasound scan) and five with no history of MO were Sanger sequenced for entire GDF9 regions (Chr14:42748507-42752672) Sequencing of GDF9 shown high level of conservativity however intronic region covers STS (submitted to NCBI, accession numbers: KX139147; KX139148; KX139149): g.42750467GT[7]GA[9], g.42750467GT[8]GA[11], g.42750467GT[8]GA[12]. None of them were claimed as functional according to ovulation rate. Thus we genotyped this STS on sequencer CEQ8000 Genetic Analysis System by Fragment Analysis application (Beckman Coulter) in mares grouped by breed Arabians (n = 73) and Thoroughbreds (n = 53). Genotype frequencies were estimated for two breeds and were compare using a Chi 2 test. The analysis indicate that none of the haplotype frequencies were significant in any breeds so we could suppose that variation in BMP15 GDF9 are not linked with multiple ovulation in horses. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Pregnancy-induced changes of leptin concentration in Polish Longwool ewes Małgorzata Szczęsna, Katarzyna Kirsz, Dorota A. Zięba Department of Animal Biotechnology, University of Agriculture in Krakow, Poland e-mail: m.szczesna@ur.krakow.pl Leptin is an adipose-derived hormone characterized by wide spectrum of actions, particularly in the regulation of food intake, energy homeostasis and reproduction. Pregnancy is a state of physiological hyperphagia, which is an adaptive response that facilitates energy storage in preparation for the high metabolic demands of gestation and subsequent lactation. The aim of this study was to investigate the leptin secretion depending on the stage of pregnancy in Polish Longwool sheep. The experiments were performed on 4 pregnant ewes. Females were 3 to 4 years old, weighed 60 ± 5 kg, and were fed ad libitum. Ewes were housed under natural photoperiodic and thermoperiodic conditions (19°57’E, 50°04’N) and were mated in October. Blood samples (5 ml) were collected at 15-min intervals beginning at dusk and were continued for 4 hrs under the red light. Experimental treatments were repeated at 2-weeks intervals starting before mating (–15 d.g.; day of gestation) and continuing between 30 and 135 day of pregnancy. Leptin concentration in the plasma were determined using RIA. Results of experiments provided that leptin concentration increased during the first half of pregnancy and after peaking at day 60 (4.01 ± 0.30 ng/ml), it began to decline. Concentration of leptin observed in periparturient ewes at 135 d.g. (1.44 ± 0.13 ng/ml) was comparable to concentration noticed before mating (1.42 ± 0.09 ng/ml) and during the first month of pregnancy (1.21 ± 0.09 ng/ml). These findings confirm that, in seasonally breeding ewes, pregnancy-induced hyperphagia occurs despite of the high concentration of leptin in midgestation and support the hypothesis that pregnant females become resistant to the anorectic actions of leptin. Research supported by NCN 2013/09/B/NZ4/01532. 109 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Analysis of changes in the frequency of FST genotypes depending on inbreeding of native Puławska sows Magdalena Szyndler-Nędza1, Katarzyna Ropka-Molik2, Katarzyna Piórkowska2, Aurelia Mucha1, Marek Babicz3, Tadeusz Blicharski4 Department of Animal Genetics and Breeding, National Research Institute of Animal Production, Balice near Krakow, Poland 2 Department of Genomics and Animal Molecular Biology, The National Research Institute of Animal Production, Balice near Krakow, Poland 3 Department of Swine Breeding and Production Technology, University of Life Sciences in Lublin, Poland 4 Institute of Genetics and Animal Breeding of the Polish Academy of Sciences, Jastrzębiec, Poland 1 e-mail: aurelia.mucha@izoo.krakow.pl Increased inbreeding in a population has a negative effect mainly on reproductive and health traits. In small populations, it is recommended to monitor the inbreeding and genetic variation based on the polymorphism of marker loci alleles. Follistatin, a cytokine isolated from the ovarian follicular fluid of pigs, has multiple activities. Blowe et al. [2006], who underlined the physiological role of follistatin, proposed the FST gene as a marker of reproductive traits in sows. The aim of the study was to analyse the effect of the inbreeding coefficient on changes in the frequency of FST genotypes (follistatin; 16q14) in a small number native Puławska breed. Based on pedigree analysis, it was classified into five groups according to the coefficient of inbreeding F: (F < 0.008 (1); 0.008 ≤ F < 0.0313 (2); 0.0313 ≤ F < 0.0625 (3); 0.0625 ≤ F < 0.098 (4); F ≥ 0.098 (5)). A total of 131 sows were chosen from the groups for further study and samples of biological material (hair bulbs) were collected from the animals. Most of the analysed animals had the homozygous genotype FST BB (59.38%), followed by heterozygous FST AB (30.47%) and FST 110 AA genotypes (10.16%). The chi-square test based on Hardy-Weinberg equilibrium demonstrated that the studied group of sows is not in genetic equilibrium (P = 0.0496). Detailed analysis of the changes in FST genotype frequency in different inbreeding groups showed the lack of genetic equilibrium in group two (0.008 ≤ F < 0.0313, P = 0.0155) and above all in group five which had the highest inbreeding coefficient (F ≥ 0.098, P = 0.0000). A relationship was also observed between the inbreeding coefficient and the frequency of FST genotypes (Spearman correlation). As the inbreeding coefficient increased to F < 9.8%, the number of animals with FST BB genotype increased significantly (r = 0.199; P ≤ 0.05). However, when the inbreeding value of sows exceeded F ≥ 9.8%, the number of animals with FST BB and FST AB genotypes decreased. In summing up the results, it is concluded that the increase in the inbreeding coefficient of Puławska sows beyond F ≥ 9.8% was related to a reduced number of sows with the FST B allele (FST BB and FST AB) in their genotype. Analiza zmian frekwencji genotypów genu FST w zależności od zinbredowania loch rodzimej rasy puławskiej Magdalena Szyndler-Nędza1, Katarzyna Ropka-Molik2, Katarzyna Piórkowska2, Aurelia Mucha1, Marek Babicz3, Tadeusz Blicharski4 Dział Genetyki i Hodowli Zwierząt, Instytut Zootechniki – PIB w Balicach k. Krakowa Dział Genomiki i Biologii Molekularnej Zwierząt, Instytut Zootechniki – PIB w Balicach k. Krakowa 3 Katedra Hodowli i Technologii Produkcji Trzody Chlewnej, Uniwersytet Przyrodniczy w Lublinie 4 Instytut Genetyki i Hodowli Zwierząt PAN w Jastrzębcu 1 2 e-mail: magdalena.szyndler@izoo.krakow.pl Wzrost wartości inbredu w populacji wpływa negatywnie przede wszystkim na cechy związane z rozrodem i zdrowotnością. W małych populacjach zalecane jest monitorowanie inbredu oraz zmienności genetycznej na podstawie wskaźników polimorfizmu alleli loci markerowych. Folistatyna należy do cytokin, wydzielonych z płynu pęcherzykowego jajników świni, o wielokierunkowym oddziaływaniu. Blowe i inni [2006], podkreślając fizjologiczną rolę folistatyny, zaproponowali gen FST jako marker cech rozpłodowych loch. Celem pracy była analiza wpływu wartości współczynnika inbredu na zmianę frekwencji genotypów genu FST (follistatin; 16q14) u mało licznej rodzimej rasy puławskiej świń. W oparciu o analizę rodowodową podzielono ją na pięć grup o określonej wartości współczynnika inbredu F (F < 0,008 (1); 0,008 ≤ F < 0,0313 (2); 0,0313 ≤ F < 0,0625 (3); 0,0625 ≤ F < 0,098 (4); F ≥ 0,098 (5)). Z wyznaczonych grup wytypowano do dalszych badań 131 loch, od których pobrano materiał biologiczny – cebulki włosowe. U analizowanych osobników najwięcej miało genotyp homozygotyczny FST BB (59,38%), następ- nie heterozygotyczny FST AB (30,47%) i FST AA (10,16%). Analiza Ch2 zgodnie z prawem Hardy’ego-Weinberga wykazała, że objęta badaniami grupa loch nie jest w równowadze genetycznej (P = 0,0496). Analizując szczegółowo zmiany frekwencji genotypów FST w poszczególnych grupach inbredu, stwierdzono brak równowagi genetycznej w grupie drugiej (0,008 ≤ F < 0,0313, P = 0,0155) i przede wszystkim w grupie piątej o najwyższej wartości współczynnika inbredu (F ≥ 0,098, P = 0,0000). Wykazano również zależność między wartością współczynnika inbredu a frekwencją genotypów genu FST (korelacja Spearmana). Wraz ze wzrostem współczynnika inbredu (do wartości F < 9,8%) istotnie wzrastała liczba osobników o genotypie FST BB (r = 0,199; P ≤ 0,05). Jednak przekroczenie wartości inbredu loch F ≥ 9,8% skutkowało zmniejszeniem liczby osobników o genotypie FST BB i FST AB. Podsumowując uzyskane wyniki, można stwierdzić, że wzrost wartości współczynnika inbredu loch rasy puławskiej powyżej F ≥ 9,8% związany był ze zmniejszeniem liczby loch posiadających w genotypie allel FST B (FST BB i FST AB). 111 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Extract from pine needle enriched with L-arginine prevents the protein malnutrition of American mink (Neovison vison) kits Aleksey R. Unzhakov1, Nikolay N. Tyutyunnik1, Svetlana N. Sergina1, Nadezhda P. Chernobrovkina2, Ekaterina P. Antonova1, Lyudmila B. Uzenbaeva1, Irina V. Baishnikova1, Viktor A. Ilyukha1, Stanisław Łapiński3, Piotr Niedbała3 Institute of Biology, Karelian Research Centre, Russian Academy of Science, Petrozavodsk, Russia Forestry Research Institute, Karelian Research Centre, Russian Academy of Science, Petrozavodsk, Russia 3 Institute of Animal Sciences, University of Agriculture in Krakow, Poland 1 2 e-mail: uar@bio.krc.karelia.ru Protein malnutrition leads to metabolic changes which are accompanied by a reduced growth rate in kits and a restriction of the biotic potential of an organism. This might be of particular importance for strict carnivores, for example, cat and mink (Neovison vison) because of their higher protein requirements than in other domestic mammals. Prevention or treatment of mammals’ protein malnutrition during the early postnatal period of life might be achieved by the supplementation of the diet with L-arginine. A search for the new sources of this essential amino acid led to use the extract from pine needle enriched with L-arginine in mink diet. Moreover, pine needles are the source of numerous vitamins, microelements and also 18 free amino acids. This study aimed to investigate the effect of extract from pine needle enriched with L-arginine on weight gain and protein metabolism in hypotrophic mink kits during the early post-weaning period of rapid growth. A total of 20 two-months-old mink kits of the standard dark-brown genotype with signs of hypotro- 112 phy were used. They were divided into two groups: control and experimental. The control animals were fed the basal diet and the experimental animals were fed the basal diet supplemented with 5 ml extract from pine needle containing 7.5 mg L-arginine / kg body weight for 14 days. Weight gain was higher and serum transaminases’ activities were lower (AST – by the 23.3%, ALT – by the 26.0%) in mink kits of experimental group compared with those of control one. Decrease in enzymes’ activities indicates an improvement in the hepatic function and protein metabolism. In conclusion, feeding mink kits with the extract from pine needle enriched with L-arginine, increased their viability by 50–60%. The study was carried out under state order (projects № 0221-2014-0001 and № 01201257867) and partially of the grant for Russian-Polish Interacademic Cooperation. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Glutathione peroxidase activity, respiration intensity and spermatozoa survival in bull ejaculates Vasyl Vlizlo, Natalia Kuzmina, Dmytro Ostapiv, Mykola Sharan, Iryna Yaremchuk, Julia Bodnar Institute of Animal Biology NAAS, Lviv, Ukraine e-mail: m_sharan@ukr.net The aim was to study correlation between activity of glutathione peroxidase, oxygen consumption intensity and spermatozoa survival in bull ejaculates. For experiments, freshly obtained bull ejaculates (n = 48), with volume 2–5 ml, concentration 0.8–1.2 × 109 cells, and quantity of moving spermatozoa – 75–80% were used. In ejaculates were studied: respiration intensity – polarographycally in phosphate-saline buffer Dulbecco, activity of glutathione peroxidase (GPx) – with Elman reagent and spermatozoa survival at temperature 2–4°С (hours). Statistical analysis of the experimental results was conducted by the Student-t test using the correlation ratio (η). Bull ejaculates characterize by GPx activity 0.79 ± 0.092 µmol GSH/min × mg of protein, respiration intensity 8.1 ± 0.6 ng-atom О/min × 0.1 ml of semen and spermatozoa survival at temperature 2–4°С 114 ± 31.8 hours. It was determined, that when activity of GPx was more than 0.80 µmol GSH/min × mg of protein, semen respiration intensity was highest – 9.4 ± 0.30 ng-atom О/min × 0.1 ml of semen. It decreased on 28.7% (p < 0,01), when GPx activity was 0.60 µmol GSH/min × mg of protein. The correlation between enzyme activity and oxygen consumption was – η = 0.298. Addition of inhibitors of mitochondrial respiratory chain did not alter the correlation between respiratory intensity and GPx activity: high oxygen consumption after addition of sodium fluoride (η = 0.432), amital (η = 0.332), sodium azide (η = 0.215) and Na2EDTA (η = 0.289) was manifested when enzyme activity was over 0.80 µmol GSH/min × mg of protein and had lower value when GPx activity was less than 0.80 µmol GSH/min × mg of protein. The highest spermatozoa survival (123 ± 8.1 hours) was registered when GPx activity was more than 0,80 µmol GSH/min × mg of protein, whereas when enzyme activity was less than 0,60 µmol GSH/min × mg of protein, spermatozoa survival was lowest (93 ± 8.6 hours; p < 0.001). The correlation between enzyme activity and survival of spermatozoa equalled – η = 0.416. 113 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The evaluation of the ability to differentiate of putative stem cells from mature pig ovaries Kamil Wartalski, Gabriela Gorczyca, Marta Hereta, Małgorzata Duda Department of Endocrinology, Institute of Zoology, Jagiellonian University in Kraków, Poland e-mail: kamil.wartalski@doctoral.uj.edu.pl The ovaries are organs in which the presence of stem cells has not yet been indisputably proven. Mature ovaries may contain adult stem cells (ASCs). ASC is a pool of cells that has an unknown ability to differentiate. According to one hypothesis, ASCs may be the source of cancer stem cells (CSCs) by the accumulation of genetic mutations. There is a great similarity of morphological and functional stem cells to cancer cells, which requires explanation. This will allow to understand the etiology of ovarian cancers and processes such tumor initiation and its growth. This is important because ovarian tumors are the second most common in women after breast cancer. In order to obtain a pool of putative stem cells (PSCs) the magnetic beads insulated with epitope SSEA-4 were used. The obtained cells were cultured in vitro for 7 and 14 days in medium DMEM/F12 with the addition of SCF (stem cells factor) and special supplement B-27. 114 During the culture, the observed characteristic clusters containing ASCs were observed. Cells in clusters showed immunolocalization of proliferation markers such as Ki-67 and PCNA. We also showed the presence of protein markers for stem cells such as Oct-3/4, c-Kit, Sox2 and GDF-9 protein associated with the development of primary follicles. These results were obtained both on paraffin sections of the ovary and cells from culture. Similar results were obtained in control tissue – neonatal porcine ovaries. At the protein level we were able to demonstrate the expression of Ki-67; PCNA; c-Kit and GDF-9. Loading control constituted b-actin. The transcript level also revealed the presence of GDF-9. It also indicated expression of c-Kit receptor, and Ki-67. This work was supported by a grant K/DSC/002993. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Characterization and possible involvement of mesenchymal stem cells in endometrium of porcine uterus model Jerzy Wiater, Janusz Karasiński 1 Department of Cell Biology and Imaging, Jagiellonian University in Kraków, Poland e-mail: jerzy.wiater@doctoral.uj.edu.pl Stem cells are characteristic for various organs, which have high potential for remodelling and regeneration. The uterus is an organ with outstanding ability to regenerate its complex tissue structures. It is composed of two main compartments: the endometrium and myometrium. The endometrium undergoes about 400 cycles of shedding, proliferation and differentiation during female reproductive period. Mesenchymal stem cells (MSCs) are self-renewing and multipotent cells, which may differentiate into more than one cell type. We have shown the presence of stem or progenitor cells in the porcine endometrium by demonstrating their in vitro differentiation into adipogenic and osteogenic lineages and expression of some mesenchymal stem cell markers by RTPCR. Hence, we aim to further characterize these cells and study their potential to differentiate into various lineages. Experiments were performed on cryostat sections and in vitro cultured cells from porcine uteri. For characterization and localization of MSCs markers we used Western blot and immunohistochemistry meth- ods, and antibodies against MSCs markers CD29, CD44, CD90, CD105, CD140b, against vimentin, as well as markers for proliferating cells PCNA and Ki67, and pluripotency marker Oct-3/4. Cells cultured in vitro expressed MSCs markers such as: CD29, CD44, CD105, CD140b and vimentin, as well as marker for proliferating cells PCNA. In tissue sections immunofluorescent signals from MSCs were localized in the endometrium close to the endometrial glands and blood vessels which may suggest their potential involvement in gland formation and angiogenesis. Western blot analysis showed the presence of CD44, CD105, CD140b, vimentin, PCNA and Oct-3/4 signals. Our results indicate that MSCs or progenitor cells may be responsible for regeneration of endometrial tissue structures such as luminal, glandular and vascular epithelium. This work was supported by grant from the National Science Centre: NN 303 804 240. 115 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Pig with reduced expression of the Galα(1,3)Gal epitope as a potential donor of skin grafts – preliminary study Jerzy Wiater1, Janusz Karasiński1, Jacek Jura2, Zdzisław Smorąg2, Ryszard Słomski3, 4, Marek Romek1 1 2 Department of Cell Biology and Imaging, Institute of Zoology, Jagiellonian University in Kraków, Poland Department of Animal Reproduction Biotechnology, National Research Institute of Animal Production, Balice near Krakow, Poland 3 Institute of Human Genetics of the Polish Academy of Sciences, Poznan, Poland 4 Department of Biochemistry and Biotechnology, Poznań University of Life Sciences, Poznan, Poland e-mail: jerzy.wiater@doctoral.uj.edu.p Xenotransplantation of the living organs as well as individual tissues or cells from pig to human is a promising method which will increase the number of available grafts and shorten the waiting time for a transplant. The organ size and physiology of the pig is almost identical to human. Some animal materials are already used in humans, such as biotechnology dressing of skin from transgenic pigs, but these are treated so that they contain no living cells. The major difficulty in xenotransplantation is high risk of hyperacute rejection of xenograft so reducing the expression of the Galα(1,3)Gal by the transgenesis may solve this problem. Therefore, the aim of this study was to determine the level of expression of Galα(1,3) Gal epitope in skin from transgenic pig. In our experiments we used skin obtained from pigs designed to express human: α-galactosidase (αGal), α1,2-fucosyltransferase (α1,2FT) and αGal × α1,2FT. Experiments were performed on cryostat sections and in vitro cultured keratinocytes and fibroblasts. For localization and determination of ex- 116 pression level of Galα(1,3)Gal we used lectin BS-IB4 conjugated with AlexaFluor 647 flourochrome, while for αGal and α1,2FT specific antibodies against this proteins. The fluorescence intensity was analysed with confocal microscope LSM 510 META Axiovert 200 M (Carl-Zeiss, Germany). Our results show that the level of Galα(1,3)Gal expression in the skin of transgenic pigs is lower compared to control non-transgenic porcine skin. The lowest expression of Galα(1,3)Gal was recorded in skin from double transgenic pig (αGal × α1,2FT). Moreover, there were no differences between keratinocytes and fibroblasts cultured in vitro. In conclusion skin of transgenic pig with living cells may be a promising material for xenotransplantation. This work was supported by grant from National Center for Research and Development No. INNOMED/I/17/ NCBR/2014 and grant No. K/DSC/002994 from Jagiellonian University in Kraków. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The changes of CRP and IL-10 secretion from piglet lymphocytes in response to adiponectin Joanna Zubel-Łojek, Anna Latacz, Ewa Ocłoń Department of Animal Physiology and Endocrinology, University of Agriculture in Krakow, Poland e-mail: e.oclon@ur.krakow.pl It is now known that adipose tissue is a dynamic endocrine organ, secreting a number of biologically active proteins, also known as adipokines. These peptides could participate in the development of obesity-related complications through regulation of inflammatory responses. Most adipokines are proinflammatory. In contrast, adiponectin exerts an anti-inflammatory effect and the actions of this protein involve a variety of mechanisms that are poorly defined. The present study was carried out to examine the in vitro influence of adiponectin addition on the cytokines (CRP and IL-10) lymphocytic secretion in piglets during inflammation. Studies were performed on 10 – week old piglets (Polish Landrace, n = 24), which were divided into control group (I) and three experimental groups; II (with obesity), III (with acute inflammation) and IV (with obesity and acute inflammation). Peripheral blood lymphocytes (PBL) were isolated from fresh heparinized piglet blood using standard gradient sedimentation technique. Isolated lymphocytes were placed in medium (RPMI 1640, 10% fetal calf serum, L-glutamine, penicillin, streptomycin) and cultured for 72 h with or without 1nM of adiponectin. The enzyme-linked immunosorbent assay method was used for estimation of CRP and IL-10 in culture supernatant. The results obtained indicated that CRP lymphocytic secretion depends on physiological condition and significantly increased during inflammation and obesity, whereas secretion of IL-10 decreased only in obese animals (p < 0.05). In addition, adiponectin in vitro supplementation inhibits CRP (from 36% to 70%) and stimulates (23‒492%) IL-10 secretion (p < 0.05). To summarize, obtained result show that adiponectin has a direct anti-inflammatory impact on the piglet lymphocytes. Supported by: grants from NCN (NN 311 227 138) and DS (3243/KFiEZ). 117 Session 3 ”7th Poultry Days” – current problems of poultry and ornamental bird breeding International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The effect of the humic acid and herbal additive supplement on production and carcass parameters of broiler chicken Veronika Pistová, Henrieta Arpášová, Cyril Hrnčár Department of Poultry and Small Animal Husbandry, Faculty of Agrobiology and Food Resources, Slovak University of Agriculture, Slovakia e-mail: xpistova@uniag.sk In this study the effect of humic acids and dietary herbal additive: clove (Syzygium aromaticum), lavender (Lavendula angustifolia) and black pepper (Piper nigrum L.) on production and carcass parameters of Ross 308 broiler chicken (n = 60) were studied. The chicken of the control group were fed with complete feed mixtures without any additives. Chicken in treatment T1 were fed a diet containing 1% of humic acid and drank a water containing 150 mg/l of herbal additive. Chicken in treatment T2 were fed with complete feed mixture without any additives and drank a water containing 150 mg/l of herbal additive. From production parameters the body weight, feed intake and feed conversion were observed. From carcass parameters the carcass weight, weight of heart, liver, gizzard and carcass yield were evaluated. The results shout that the body weight was significantly higher (P ≤ 0.05) in treatments groups compared to the control group (the order of the groups: 1796.6 ± 188.2; 2053.1 ± 197.8 and 2140.2 ± 300.5 g ± SD). The feed intake was in the control group 3.10 kg, in the treatment T1 3.00 kg and in the treatment T2 3.13 kg. The carcass weight was in the treatment T2 significantly higher (P ≤ 0.05) in compare with control group (values in the order of the groups: 1293.8 ± 179.5; 1428.9 ± 156.5 and 1532.6 ± 283.1 g ± SD). In the treatment T1 was weight of gizzard (44.7 ± 7.6 g ± SD) significantly higher (P ≤ 0.05) compared to control group (35.5 ± 5.8 g ± SD). In the treatment T2 was weight of liver (74.1 ± 12.5 g ± SD) significantly higher (P ≤ 0.05) compared to control group (50.1 ± 11.5 g ± SD). In conclusion, supplement by humic acid and herbal additive can improve production parameters of broiler chicken. 121 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 In ovo technology – efficient method of chicken microbiome stimulation Maria Siwek1, Anna Sławińska1, 2, Katarzyna Stadnicka1, Aleksandra Dunisławska1, Arkadiusz Płowiec1, Giuseppe Maiorano2, Marek Bednarczyk1 1 University of Science and Technologies, Bydgoszcz, Poland 2 University of Molise, Campobasso, Italy e-mail: siwek@utp.edu.pl Digestive tract of animals harbors living and metabolizing microorganisms. They influence on physiological functions of the host and are considered fundamental drivers of some physiological traits, including metabolism and immune system. In EU ban on antibiotics in 2006 directed interests of the poultry producers to use natural growth promoters (NGPs), such as prebiotics, probiotics and synbiotics. These NGPs stimulate colonization of the chicken intestines with beneficial microflora. In poultry farming, chicks are given NGPs at first day after hatching (in-feed or inwater). The early timing of guts colonization with commensal bacteria is crucial, otherwise they tend to get colonized by pathogens. For this purpose, we have optimized an in ovo technology for delivery of NGPs to the chicken embryo at 12th day of egg incubation. We have tested several prebiotics (e.g. inulin, GOS – galactooligosaccharides and RFO – raffinose family oligosaccharides), probiotics (e.g. Lactococcus spp., Lactobacillus spp.) and synbiotics (e.g. RFO + Lacto- 122 coccus lactis subsp. cremoris) in combination with in ovo technology. All of them proved to have beneficial effects on egg hatchability, chicken mortality rate, on broiler chicken growth and meat quality traits. There are clear differences between effects of different NGPs delivered in ovo on the host. These differences were determined based on gene expression level, bacteriology, histology of the gastrointestinal track, physiological, immunological and biochemical parameters. In conclusion, NGPs injected in ovo are beneficial for the host but their effect strictly depends on the bioactive compound used; they might act towards improvement of metabolic or immune traits. Detailed results will be discussed during presentation. Research projects partially funded by National Science Centre in Cracow, Poland (UMO-2013/11/B/ NZ9/00783) and Ministry of Education, Universities and Research in Rome, Italy (RBSI14WZCL) International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of lavender essential oil (Lavandula angustifolia) on production indicators of broiler chicken Michalina Adaszyńska-Skwirzyńska1, 2, Danuta Szczerbińska2 Poultry Farm Michalina Adaszyńska-Skwirzyńska, Żabówko, Poland Department of Poultry and Ornamental Bird Breeding, West Pomeranian University of Technology, Szczecin, Poland 1 2 e-mail: madaszynska@zut.edu.pl Currently the use of natural biologically active substances is promoted in many branches of industry and in agriculture, e.g. those with the action similar to withdrawn antibiotic growth stimulants. For instance, the properties of lavender oil can be used in animal production. The aim of this study was to estimate the effect of an addition of the lavender essential oil (Lavandula angustifolia) to drinking water on the production indicators. The study was carried out on a commercial poultry farm and experimental animals were broiler Ross 308 chickens. A total of 300 of one-day chicks were divided into three experimental groups: I group (control) – 100 specimens (water without addition of essential oil); II group – 100 specimens (addition of oil to water at a concentration of 0.2 cm3 L-1; III group – 100 specimens (addition of oil to water at concentration of 0.4 cm3 L-1). Commercial lavender oil with antimicrobial properties was administered throughout the rearing cy- cle. Water with oil was available for chicken for 6 h/ day. Chickens were fed ad libitum with mixed feeds delivered from the Feed-Processing Plant in Wałcz (PZZ Wałcz). Birds were provided with standard conditions of lighting and temperature according to the guidelines of the chick supplier. The chicken body weight and feed consumption were controlled during the experiment. To control the body weight, the chickens were weighed six times: on 1, 7, 14, 28, 35 and 42 day of fattening. Based on the carried out study it was stated that an addition of oil is characterized by a positive effect on body weight gains in broilers. The highest mean weight was recorded in the group of chickens that were given water with an addition of lavender oil (group III – oil concentration of 0.4 cm3 L-1). In this case the weight gain in relation to the control group at the end of rearing amounted to 3.7%, and the feed conversion rate (FCR) was 1.26. In the same group also the lowest mortality rate was recorded (1.0%). 123 Wpływ olejku lawendowego (Lavandula angustifolia) na wskaźniki produkcyjne kurcząt broilerów Michalina Adaszyńska-Skwirzyńska1, 2, Danuta Szczerbińska2 Ferma Drobiu Michalina Adaszyńska-Skwirzyńska, Żabówko Zakład Hodowli Ptaków Użytkowych i Ozdobnych, Zachodniopomorski Uniwersytet Technologiczny w Szczecinie 1 2 e-mail: madaszynska@zut.edu.pl Obecnie w wielu gałęziach przemysłu i w rolnictwie promuje się stosowanie naturalnych substancji biologicznie czynnych, np. o działaniu zbliżonym do wycofanych antybiotykowych stymulatorów wzrostu. Właściwości olejku lawendowego mogą zostać wykorzystane w produkcji zwierzęcej. Celem badań było określenie wpływu dodatku do wody pitnej olejku eterycznego z lawendy lekarskiej (Lavandula angustifolia) na wskaźniki produkcyjne. Badania przeprowadzono na fermie komercyjnej (Żabówko), a zwierzętami doświadczalnymi były kurczęta brojlery Ross 308. Jednodniowe pisklęta, w liczbie 300 osobników, przydzielone zostały do trzech grup doświadczalnych: I grupa kontrolna – 100 osobników (woda bez dodatku olejku eterycznego), II grupa – 100 osobników (dodatek do wody olejku w stężeniu 0,2 cm3 L-1, III grupa – 100 osobników (dodatek do wody olejku w stężeniu 0,4 cm3 L-1). Komercyjny olejek lawendowy o właściwościach przeciwdrobnoustrojowych był podawany przez 124 cały okres odchowu. Woda z olejkiem była dostępna dla kurcząt przez 6 h/dobę. Kurczęta żywiono bez ograniczeń mieszankami paszowymi dostarczanymi z Wytwórni Pasz w Wałczu (PZZ Wałcz). Ptaki miały zapewnione standardowe warunki oświetlenia oraz temperatury zgodne z wytycznymi dostawcy piskląt. W czasie trwania doświadczenia kontrolowana była masa ciała kurcząt oraz spożycie paszy. W celu kontroli masy ciała kurczęta ważono sześciokrotnie: w 1., 7., 14., 28., 35. oraz 42. dniu tuczu. Na podstawie przeprowadzonych badań stwierdzono, że dodatek olejku ma pozytywny wpływ na przyrosty masy ciała brojlerów. Najwyższą średnią wagę odnotowano w grupie kurcząt, którym podawano wodę z dodatkiem olejku lawendowego (grupa III – stężenie olejku 0,4 cm3 L-1). W tym przypadku wzrost masy w stosunku do grupy kontrolnej na koniec chowu wyniósł 3,7%, a współczynnik konwersji paszy (FCR) 1,26. W tej samej grupie odnotowano także najniższy wskaźnik śmiertelności (1,0%). International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effects of fumonisin contaminated diet on glutathione redox system and lipid peroxidation processes in broiler chickens Krisztián Balogh, Nikolett Tokodi, Erika Zándoki, Judit Szabó-Fodor, Csilla Pelyhe, Andrea Bócsai, Miklós Mézes Department of Nutrition, Faculty of Agricultural and Environmental Sciences, Szent István University, Hungary e-mail: balogh.krisztian@mkk.szie.hu Fumonisins are secondary metabolites of the corn pathogen fungus Fusarium verticillioides. Based on their structural similarities the different fumonisins are clustered into four groups. In feedstuffs and feeds the fumonisin B1 and B2 (FB1 and FB2) are the most frequent. They alter the sphingolipid metabolism in animals and they also have carcinogenic effects. The purpose of our study was to investigate the effect of feed-borne fumonisin (FUM) exposure on the amount and activity of glutathione redox system and on the lipid peroxidation processes of broiler chickens. A total of 68 3-week old broiler cockerels were divided into two groups: a control and a FUM treated one. The control group was fed broiler commercial grower diet, while the treated group received the same diet artificially contaminated with fumonisins (30 mg FB1 + FB2/kg feed) for 14 days. Samplings (n = 6/group) were done at day 1, 2, 3, 7 and 14 of mycotoxin exposure. At every sampling blood and post mortem liver, spleen and kidney samples were taken, in which the parameters of glutathione redox system (reduced glutathione (GSH) concentration and gluathione-peroxidase (GPx) activity) and also the meta-stable end product of the lipid peroxidation processes, the malondialdehyde (MDA) concentration were measured. GSH concentration in the spleen of the treated group was significantly higher at day 1, 2 and 7 of sampling. The increased amount of co-substrate resulted in elevated GPx activity at day 2, 3 at 7 of FUM exposure as compared to the control. In kidney samples FUM treatment increased the GSH concentration and GPx activity in kidney at day 3 of the experiment. Lower GSH concentrations were measured at days 2, 3 and 7 in blood plasma, which led to an increase in lipid peroxidation processes as measured by the MDA concentrations. The study was supported by the János Bolyai Research Scholarship (BO/261/13) of the Hungarian Academy of Sciences and by the OTKA (PD-104823). 125 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Seasonal changes in the behavior of emu (Dromaius novaehollandiae) under farm conditions Mateusz Bucław Department of Poultry and Ornamental Bird Breeding, West Pomeranian University of Technology, Szczecin, Poland e-mail: mateusz.buclaw@zut.edu.pl The year season strongly affects the behaviour of emus, which is a source of unique problems in their adaptation to the farm environment. While creating the farming environment for any livestock animals, the specificity of their behaviour should be taken into account, since this has an impact on the animal welfare and the productivity of the herd. The aim of this study was to present selected elements of the emu behavior during the off breeding season, the breeding preparation period and the breeding season. The study focuses on feed, water and sand intake, plumage care, resting, agonistic behavior, and other activities, mostly standing still, aimless walking or observing the surrounding. The troop consisted of 11 emus (5 females and 6 males) aged 12 years, housed in a free-ranging system. Observations were carried out in three, 3-weeklong stages, during which 60-minute watching sessions took place on 3 randomly chosen days. 126 A significant effect was found of each season on the behavior patterns of emu related to feeding, pecking, sand intake, plumage care, resting, and agonistic behavior. Plumage care and agonistic behavior took most time during the off breeding season. The longest periods of resting, feeding and sand intake were observed during the breeding preparation period, which suggests that birds prepared for the breeding season by saving energy and gathering reserve material in the body. Feeding and water drinking behavioural patterns were similar in all the seasons. Most of the time, regardless of the period of observation, the emu spent on such activities as standing still, walking aimlessly, watching the sourrounding area, which are affected by their housing system. The emu belongs to birds whose behavior is considerably affected by the season. Seasonality and the resulting changes in behavior are associated with reproduction, which is superior to the remaining activities. Sezonowe zmiany w zachowaniu się emu (Dromaius novaehollandiae) utrzymywanych w warunkach fermowych Mateusz Bucław Zakład Hodowli Ptaków Użytkowych i Ozdobnych, Zachodniopomorski Uniwersytet Technologiczny w Szczecinie e-mail: mateusz.buclaw@zut.edu.pl Sezon ma znaczący wpływ na zachowanie się emu, co powoduje unikatowe problemy ich adaptacji jako zwierząt hodowlanych. Specyficzne zachowania zwierząt muszą zostać uwzględnione przy tworzeniu warunków środowiskowych utrzymania, gdyż determinuje to ich dobrostan i produktywność. Celem niniejszej pracy było przedstawienie wybranych elementów zachowania się emu w okresie spoczynku, wchodzenia w sezon rozrodczy oraz sezonu rozrodczego. W pracy analizowano zagadnienia dotyczące: pobierania pokarmu, wody i piasku oraz porządkowania upierzenia, odpoczynku, zachowań agonistycznych i pozostałych, związanych głównie ze staniem w bezruchu, chodzeniem bez celu czy obserwowaniem otoczenia. Badane stado liczyło 11 ptaków (5 samic i 6 samców) w wieku 12 lat, utrzymywanych systemem otwartym. Obserwacje prowadzono w trzech trzytygodniowych etapach, w których przez losowo wybrane 3 dni prowadzono 60-minutowe obserwacje, dwa razy dziennie. Stwierdzono istotny wpływ poszczególnych sezonów na behawior związany z pobieraniem pokarmu, dziobaniem i pobieraniem piasku, porządkowaniem upierzenia, odpoczynkiem, a także na zachowania agonistyczne. Najwięcej czasu na porządkowanie upierzenia i zachowania agonistyczne emu poświęcały w okresie spoczynku. Najdłuższy czas odpoczynku oraz pobierania pokarmu i piasku stwierdzono w okresie wchodzenia w sezon rozrodczy, co sugeruje przygotowywanie się ptaków do sezonu rozrodczego poprzez oszczędzanie energii i magazynowanie w organizmie materiału zapasowego. Zachowania związane z żerowaniem i pobieraniem wody w przypadku poszczególnych sezonów były zbliżone. Najwięcej czasu, bez względu na okres obserwacji, emu poświęcały na czynności związane głównie ze staniem w bezruchu, chodzeniem bez celu czy obserwowaniem otoczenia, na które wpływa ich fermowe utrzymywanie. Emu należą do ptaków, u których sezon w znaczący sposób oddziałuje na ich zachowanie. Sezonowość i zmiany w zachowaniu są związane z rozrodem, który podporządkowuje sobie pozostałe zachowania. 127 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The content of selected chemical components in hen eggs depending on egg production cycle Jolanta Calik Department of Animal Genetic Resources Conservation, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: jolanta.calik@izoo.krakow.pl The experiment used eggs from four layer breeds covered by the genetic resources conservation programme: Greenleg Partridge (Z-11), Yellowleg Partridge (Ż-33), Sussex (S-66), Leghorn (G-99) as well as from Hy-Line Brown commercial layers, kept on a farm at the Experimental Station of the National Research Institute of Animal Production in Chorzelów near Mielec. The aim of the study was to evaluate the content of some chemical components in hen eggs depending on egg production cycle. Chemical analyses of eggs were performed at 33 and 83 weeks of age at the Institute’s Central Laboratory. The highest crude protein and ash content of albumen at 33 weeks of age were found in eggs from Hy-line Brown layers, and at 83 weeks of age in S-66, G-99 and Hy-Line Brown hens, the eggs of which also had the highest dry matter and ash content. The highest crude protein content, especially in the second year 128 of production, was noted in yolks from Z-11 and Ż-33 hens of conservation breeds. Significantly lower protein content, at both 33 and 83 weeks, was observed in yolks of eggs from G-99 and Hy-Line Brown hens. The yolks of eggs from S-66 layers had the highest concentration of fat, which tended to increase with the age of the hens. Cholesterol content ranged from 13.90 to 14.78 at 33 weeks and from 14.29 to 15.48 mg/g yolk at 83 weeks of age, with Z-11 hens producing eggs with the lowest cholesterol percentage of all breeds. The high nutrient content observed in the second production cycle suggests the appropriateness of using hens for the period of two years. This study was supported from the Multiannual Program no. 17-1.10.9 financed by the Ministry of Agriculture and Rural Development. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of caponizing Yellowleg Partridge (Ż-33) cockerels on meat quality Jolanta Calik1, Katarzyna Połtowicz2, Robert Gąsior3 1 Department of Animal Genetic Resources Conservation, National Research Institute of Animal Production, Balice near Krakow, Poland 2 Department of Animal Genetic and Breeding, National Research Institute of Animal Production, Balice near Krakow, Poland 3 Central Laboratory, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: jolanta.calik@izoo.krakow.pl Capon production has recently attracted increasing interest in Poland and across the world. According to the EU Commission Regulation No 543/2008 of 16 June 2008, “a capon is a male fowl castrated surgically before reaching sexual maturity and slaughtered at a minimum age of 140 days; after castration the capons must be fattened for at least 77 days”. The aim of the study was to determine the effect of caponization on the body weight and selected meat quality parameters of Yellowleg Partridge (Ż-33) cockerels. Ż-33 cockerels were randomly assigned to two groups with 40 birds. Group I (control) consisted of uncastrated cockerels and birds from group II were castrated at 9 wk of age. The castration was performed under local anesthesia by a veterinary surgeon. Birds were kept until 24 wk of age and fed the same diet ad libitum. Dressing percentage, proportion of breast and leg muscles, giblets and abdominal fat were de- termined postmortem. Physical characteristics of meat: pH, CIE L*a*b* colour, drip loss, cooking loss, and shear force were determined and sensory evaluation was performed. It is concluded from the results obtained that the castration of Ż-33 cockerels had a positive effect on increasing of body weight, dressing percentage and carcass muscling. Capon muscles, especially leg muscles, were characterized by better water holding capacity and tenderness, lower drip loss, lower cooking loss and better sensory scores. Therefore, capon production may provide poultry meat as a niche product for consumers who look for meat of special quality. This study was supported from the Multiannual Program no. 17-1.10.9 financed by the Ministry of Agriculture and Rural Development. 129 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Evaluation of productivity and egg quality in Sussex (S-66) laying hens during the first two years of production Jolanta Calik, Michał Puchała, Joanna Obrzut Department of Animal Genetic Resources Conservation, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: jolanta.calik@izoo.krakow.pl Due to the increasing costs of rearing resulting from increasing prices of chicks, feeds and energy carriers, keeping layers for one season may be unprofitable and it might be appropriate to prolong their use for the next two or more production periods. The aim of the study was to analyse the productivity and egg quality of Sussex (S-66) laying hens during the first two years of production. The study was performed on 200 hens. The health of the birds during the first two years of production was very good. Laying rate averaged 53.76% during the first year of production (21 to 56 weeks of age) and 48.58% in the second (63 to 99 weeks of age). The quality of eggs was assessed (30 eggs per test) at 21, 33, 43, 56, 73, 83 and 93 weeks of age. Average egg weight increased from 44.05 (week 21) to 62.89 g (week 93). The increase in egg weight was paralleled by a significant increase in 130 yolk and shell weight. Eggs showed good parameters of albumen (albumen height and Haugh units) and shell quality, but these traits deteriorated significantly towards the end of the second egg production period (83 to 93 weeks). It was established from the results obtained that keeping Sussex (S-66) hens for the second production cycle has no adverse effect on their health. The good productivity and egg quality, which persisted into the second year of production, shows that it is appropriate to use Sussex (S-66) hens for the period of two years. This study was supported from the Multiannual Program no. 17-1.10.9 financed by the Ministry of Agriculture and Rural Development. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The welfare of hens in various systems of extensive farming Magdalena Dykiel1, Zofia Sokołowicz2 Faculty of Agriculture and Rural Development, State Higher Vocational School in Krosno, Poland Department of Animal Production, Faculty of Biology and Agriculture, University of Rzeszów, Poland 1 2 e-mail: dykiel@op.pl The study presents the examined welfare of commercial hens (group I) and Greenleg Partridge hens (group II) housed in various systems of extensive farming, i.e. in the floor raising system without exercise area, free range and organic system. In 20th, 36th and 54th weeks of age, observations concerning the behavior of hens were conducted in each of the studied methods of housing system. The number of physically active hens, sitting hens, hens showing signs of aggression and “playing” hens was registered. In all periods of evaluations, both in free range and organic system, regardless of weather conditions, Greenleg Partridge hens willingly benefited from the exercise area. In the free range, 73.06% of hens stayed on the run, and about 40% of commercial hens took advantage of the run. In the organic breeding, the part of hens using the run from group I amounted to 52.22%, and from group II – 76.67%. The carried out researches showed that the participation of comfort behavior in particular farming system was as follows: 15.74% in the floor raising system, 25.05% in free range and 21.57% in organic housing system. In the free range and organic farming, symptoms of aggression were not found neither in a group of Greenleg Partridge hens nor in a group of commercial hens. In a group of hens kept without an access to run, the frequency of aggressive behaviors reached the level of 2.78% in group I and 3.61% in group II. Symptoms of aggression occurring in the floor raising system were mostly mild, but there were also cases of pecking, fighting and chasing away defeated individuals, preening and even pulling out feathers. Attacked hens responded to these actions through the cry, escape or revenge. 131 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 In ovo study on calcium protective effect against cadmium embryotoxicity Małgorzata Dżugan1, Monika Wesołowska1, Marcin Lis2, Teodora Król3, Jakub Nowak4, Jerzy W. Niedziółka2 Department of Chemistry and Food Toxicology, University of Rzeszow, Poland Department of Veterinary and Animal Reproduction and Welfare, University of Agriculture in Krakow, Poland 3 Department of Cell Biology and Electronic Microscopy, The Jan Kochanowski University in Kielce, Poland 4 Technological Incubator, Stalowa Wola, Poland 1 2 e-mail: mdzugan@ur.edu.pl The aim of this study was to determine if simultaneous calcium (Ca) supplementation of hen’s egg could reduce cadmium (Cd) embryotoxicity during in ovo experiment. On day 4th of incubation egg albumen (60 eggs/group) was injected with cadmium alone (50 nmol/egg) or in combination with calcium (5 or 25 µmol/egg). Moreover, saline and saline with 5 or 25 µmol Ca injection were used in control groups. Hatching results and embryopathological evaluation were performed. For both studied metals their distribution in tissues of 1-day old chicks was examined by LA-ICP-OES method. The tissues structure was evaluated by transmission electron microscope. The activity of N-acetyl-β-D-glucosaminidase in chicks blood, kidney and liver was tested. Administration of cadmium decreased the hatchability of eggs (35.7%) compared to the control group (52.7%). A simultaneous administration of calcium, at a dose of 5 and 25 µmol/egg, gradually improved hatching results (43.9 and 47.3%, respectively). Similarly, the accumulation 132 of cadmium in liver, kidneys and blood was limited by calcium (P < 0.05). During the combined administration of Cd and Ca (at a higher dose) the level of N-acetyl-β-D-glucosaminidase activity in chicks blood and kidney was similar to the level of the control group (P > 0.05). Ultrastructural analysis of liver and kidney cells indicated that the most sensitive to the cadmium were mitochondria, endoplasmic reticulum and lysosomal system. Moreover, when tissues of chicks exposed to the combined effect of Cd and Ca during embryogenesis were examined, stabilizing effect of Ca ions on the cell structure, in particular on mitochondria, was observed. Conclusion: the supply of Ca seems to protect against Cd-induced disorders during the development of chick embryo. Funded by: NCN project NN 304 291 140 and DS ZWRiDZ/2016. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Preliminary study on some parameters of meat quality and antioxidant defence of grey partridge (Perdix perdix) Csaba Fernye, Márta Erdélyi, Krisztián Balogh, Zsolt Ancsin, Andrea Bócsai, Miklós Mézes Department of Nutrition, Faculty of Agricultural and Enviromental Sciences, Szent István University, Hungary e-mail: fernye.csaba@gmail.com Grey partridge was one of the most significant small game in Hungary in the 1960s and 70s. Their population dropped dramatically due to the rapid changes in the agricultural management. In spite of the small amount of partridge available for the consumers these birds provide palatable red meat. However, there is no data available about the physical and chemical characteristics of partridge meat. Purpose of present study was to study some physical meat quality and antioxidant parameters in partridge meat. Grey partridge birds at 20 weeks of age were exterminated and post mortem heart, liver, spleen and kidney tissue samples were taken. For meat quality measurements the pectoral muscle of the birds was sampled. To characterize antioxidant status, malondialdehyde (MDA) content was measured in the liver, spleen, kidney and heart homogenate and in blood plasma. Reduced glutathione (GSH) content and glutathione-peroxidase (GPx) activity were measured in blood plasma and 10,000 g supernatant fraction of tissue homogenates. For meat quality analysis pH, colour and water holding capacity were determined. MDA content of the liver, heart, kidney and spleen were 10.74 ± 0.55 µmol/g, 9.83 ± 1.14 µmol/g, 11.65 ± 3.98 µmol/g and 14.54 ± 5.46 µmol/g, respectively. GSH contents of the liver, heart, kidney and spleen samples were 2.72 ± 0.16 µmol/g protein, 2.61 ± 0.25 µmol/g protein, 2.50 ± 0.21 µmol/g protein, and 2.71 ± 0.24 µmol/g protein, respectively. GPx activity of the liver, heart, kidney and spleen samples were 2.83 ± 0.45 U/g protein, 3.41 ± 0.26 U/g protein, 2.28 ± 0.16 U/g protein and 3.06 ± 0.17 U/g protein, respectively. The mean pH value of the meat samples was 5.90 ± 0.20. Considering colour, lightness was found to be 32.88 ± 2.00, while intensity of red colour was 10.49 ± 1.88, whereas yellow colour was 3.32 ± 1.01, respectively. Our results revealed, that the measured parameters are comparable to that of other poultry species, such as chicken or turkey. 133 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The infuence of rapeseed on growth performance in broiler diets Marian Foltyn1, Vojtěch Rada1, 2, Martina Lichovníková2 Výkrm Třebíč s.r.o., Czech Republic Department of Animal Breeding, Mendel University in Brno, Czech Republic 1 2 e-mail: marian.foltyn@centrum.cz The aim of this study was to determine the effect of feeding full-fat rapeseed on growth performance and feed conversion ratio in broiler diets. Single sex (male) ROSS 308 broiler chickens were used in the experiment. A total of 800 9-day-old chickens were randomly distributed among 4 dietary treatments using 2 replicate pens per treatment and 100 birds per pen. The dietary treatments consisted of the control diet (0% of rapeseed) and diets with the 8%, 12% 134 and 15% of rapeseed. The best growth performance (2708.9 ± 16.28 g) in 36 day was in group containing 12% rapeseed in the diet. The significanty (P < 0.05) worst growth performance (2590.3 ± 16.62 g) was in group feeding diet containing 15% of rapeseed. Feed conversion ratio was similar in all groups (1.61 kg/kg in groups containing 0% and 8% rapeseed and 1.59 kg/kg in groups contaning 12% and 15% rapeseed). International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The microscopic study of the conjunctiva-associated lymphoid tissue (CALT) in Indian peafowl (Pavo cristatus) Karolina Goździewska-Harłajczuk1, Joanna Klećkowska-Nawrot1, Karolina Barszcz2 Division of Animal Anatomy, Department of Animal Physiology and Biostructure, Wrocław University of Environmental and Life Sciences, Poland 2 Department of Morphological Sciences, Warsaw University of Life Sciences, Poland 1 e-mail: k.gozdziewska.wroc@gmail.com The conjunctiva-associated lymphoid tissue (CALT) is a component of mucosa-associated lymphoid tissue (MALT), thus CALT represents a part of immune system of the body that is located in eyelids. The eyelids structure is support by lymphocyte recirculation and it forms the common conjunctival immune system, which provide immune protection of the eyeball area. The study was conducted on the 10 females of peafowl (3–4 weeks old, 1.5, 2–3 and 4 years old birds). Tissue sections were stained with haematoxylin and eosin, Masson-Goldner trichrome, azan-trichrome and methyl green-pyronin Y. The light microscopy investigation were performed with using of Zeiss Axio Scope A1 light microscope (Carl Zeiss, Jena, Germany). The obtained results showed the presence of well-developed CALT: diffuse lymphoid tissue within the connective tissue of the eyelids and the numerous lymphatic follicles mostly under the conjunctival epithelium, in all of the examined birds. The numerous lymphocytes were observed within the loose connective tissue as well as the numerous goblet cells were present within the conjunctival epithelium. The shape of goblet cells was similar as in other avian species. The examination of the CALT in Indian peafowl can be contribute to enlarge of our knowledge about the physiology and functioning of the eye and the nasopharynx in this bird species, what is very important in prevention of the respiratory system diseases in domestic, ornamental and wild birds. 135 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Monitoring of the genetic diversity of the Zatorska goose based on the pedigree information Magdalena Graczyk1, Krzysztof Andres2, Ewa Kapkowska2, Tomasz Szwaczkowski1 Department of Genetics and Animal Breeding, Poznan University of Life Sciences, Poland Department of Swine and Small Animal Breeding, University of Agriculture in Krakow, Poland 1 2 e-mail: maggra@up.poznan.pl Populations covered by the genetic resources conservation program require special monitoring of their genetic variability. It may be estimated based on the polymorphism of genetic markers , or recorded traits or pedigree information. From the economic point of view the third approach is the most desirable. It requires reliable and complete pedigree data. The aim of the study was to estimate the genetic conservation index and other parameters supporting the management of the Zatorska goose population. The analysis covered 437 of the currently living individuals of the Zatorska goose population hatched between 2012–2015. The following parameters were estimated: genetic conservation index (GCI), completeness of the pedigree (CP), inbreeding coefficient (F), coancestry coefficient (CC), genetic diversity (GD) and effective population size (Ne). A similar analysis was conducted for 5704 individuals hatched between 1990–2015. These calculations were carried out using ENDOG v4.8 program. 136 Completeness of the pedigree of the present population was high and reached 97.43% (for males: 97.92%, for females: 96.95%). Inbreeding level is low and reached 5.30% (for males: 5.56%, for females: 5.21%). The average CC was 7.13%, which means that this percentage of alleles is common for all animals. GCI is balanced for males and females and reached 3.61 and 3.59, respectively. Despite the varied number of individuals between 1990–2015, GCI has been stable (3.0–3.5) for the last 20 years with a very high concurrent genetic diversity (92.76%). Effective population size estimated on the basis of individual increase in inbreeding is 82 individuals. Comparative analysis of the estimated parameters for the whole and present population indicates stabilisation, which is beneficial from the perspective of the genetic resources conservation of the Zatorska goose. Monitorowanie zmienności genetycznej gęsi zatorskiej na podstawie informacji rodowodowych Magdalena Graczyk1, Krzysztof Andres2, Ewa Kapkowska2, Tomasz Szwaczkowski1 Katedra Genetyki i Hodowli Zwierząt, Uniwersytet Przyrodniczy w Poznaniu Katedra Hodowli Trzody Chlewnej i Drobnego Inwentarza, Uniwersytet Rolniczy w Krakowie 1 2 e-mail: maggra@up.poznan.pl Populacje objęte programem ochrony zasobów genetycznych wymagają szczególnego monitorowania zmienności genetycznej. Może być ona szacowana na podstawie polimorfizmu markerów genetycznych, rejestrowanych cech i informacji rodowodowych. Z ekonomicznego punktu widzenia trzecie podejście jest najbardziej pożądane. Wymaga ono jednak wiarygodnych i kompletnych danych rodowodowych. Celem badań było oszacowanie indeksu zachowania zmienności genetycznej i innych parametrów wspomagających zarządzanie populacją gęsi zatorskiej. Analizą objęto 437 obecnie żyjących gęsi zatorskich, wyklutych w latach 2012–2015. Oszacowano następujące parametry: indeks zachowania zmienności genetycznej (GCI), kompletność rodowodu (PC), współczynnik inbredu (F), współczynnik wspólnego pochodzenia (CC), zróżnicowanie genetyczne populacji (GD) oraz efektywną wielkość populacji (Ne). Analogiczną analizę przeprowadzono dla 5704 osobników wyklutych w latach 1990–2015. Obliczenia wykonano za pomocą programu ENDOG v4.8. Kompletność rodowodowa obecnie żyjącej populacji była wysoka i wynosiła 97,43% (u samców: 97,92%, u samic: 96,95%). Poziom inbredu jest niski i wynosi 5,30% (u samców: 5,56%, u samic: 5,21%). Średni współczynnik wspólnego pochodzenia wynosił 7,13%, co oznacza, że taki procent alleli jest identyczny dla wszystkich osobników. GCI był wyrównany dla samców oraz samic i wynosił odpowiednio: 3,61 i 3,59. Pomimo znacznych zmian w liczebności populacji w latach 1990–2015 indeks GCI od 20 lat utrzymuje się na stałym poziomie (3,0–3,5), przy jednocześnie bardzo wysokim współczynniku zróżnicowania genetycznego (92,76%). Efektywna wielkość populacji szacowana na podstawie przyrostu inbredu wynosi 82 osobniki. Analiza porównawcza oszacowań parametrów dla całej populacji i obecnie żyjącej wskazuje na stabilizację, co jest korzystne z perspektywy ochrony zasobów genetycznych gęsi zatorskiej. 137 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of biologically active substances on laying traits of Japanese quails Emília Hanusová, Anton Hanus, Cyril Hrnčár, Marta Oravcová National Agriculture and Food Centre – Research Institute for Animal Production, Nitra, Lužianky, Slovakia e-mail: hanusova@vuzv.sk The objective of this study was to compare the effect of additions of humic acids and probiotic in the diet on laying traits in Japanese quails (Coturnix japonica). The birds involved in experiment were farmed in the National Agriculture and Food Centre – Research Institute for Animal Production Nitra (NAFC – RIAP Nitra). Birds were housed in four-stage cage technology. Feed mixture contained 11.7 MJ ME and 200.0 g crude protein throughout the experiment. Feed and water were given ad libitum. The design consisted of one control group and two experimental groups. In the first experimental group, birds were added Humac Natur (min. 62% humic acids in dry matter) at a dose of 3 g per 1 kg of feed mixtures. In the second experimental group, birds were added probiotic based on Bacillus subtilis (min. 1 × 107 CFU) and Lactobacillus paracasei (min. 1 × 107 CFU) at a dose of 1 g per kg of feed mixtures. Daily laying traits were 138 observed during 215 days (7 months) in each group. The best laying traits were observed from 12 to 20 weeks of age in each group. In the first part of the experiment duration (July to October), birds of the first experimental group with the Humac Natur addition had the best laying traits; in the second part of the experiment duration (November to January), birds of the control group had the best laying traits. Laying traits evaluated throughout the whole experiment were better in birds of the first experimental group added Humac Natur addition in comparison to birds of the second experimental group added probiotic. Laying traits in birds of the control group were also better than these in bird of the second experimental group (added probiotic). Research indicates that birds added Humac Natur are better laying traits than birds added probiotic. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Single nucleotide polymorphism of PMCA genes in ISA Brown laying hens Eliska Horecka, Cenek Horecky, Lenka Kovarikova, Ales Knoll, Ales Pavlík Mendel University in Brno, Czech Republic e-mail: pavlik@mendelu.cz The experiments were performed in 110 ISA BROWN egg production hens, kept from 15 to 26 week of age in enriched (furnished) housing technology. Therefore, the present objective was to investigate the presence of SNPs of ATP2B1 gene. Plasma membrane calcium-transporting ATPase 1 gene (ATP2B1) in hens is located on chromosome 1, region 43 273 706 – 43 305 815 bp. ATP2B1 gene has 21 exons. In this study three exons were genotyped. In the investigated group of animals only allele without deletion in exon 10 and only allele A in exon 12 were found. In exon 8 only genotypes CC/CC, TT/CC and TT/TT were found. 139 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The effect of supplementation of enzyme preparation on base of pepsin on performance and carcass characteristics of Japanese quail Cyril Hrnčár, Denis Porubský, Ján Weis, Henrieta Arpášová, Veronika Pistová, Jozef Bujko Department of Poultry Science and Small Animal Husbandry, Slovak University of Agriculture in Nitra, Slovakia e-mail: cyril.hrncar@uniag.sk The objective of this study was to investigate the effects of three levels of supplement prepared on the base of pepsin (0.8 ml, 1.0 and 1.2 ml) in 1l of drinking water on performance and carcass characteristics of Japanese quails. Growth performance traits including weight gain, feed consumption and mortality were recorded. At the end of the experiment, five birds per treatment were slaughtered and carcass characteristics were measured. The results of this study indicated that supplementation of enzyme beneficial af- 140 fected body weight, feed consumption and mortality in quails. All carcass traits, except for abdominal fat weight, were not influenced by enzyme supplementation. The abdominal fat was higher in control birds. We found that the addition of enzyme in 1.2 ml per 1 liter of water affected growth potential of the Japanese quail the most. It is concluded that enzyme can be successfully incorporated in nutrition of Japanese quail as a growth promoter. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The adsorbents of mycotoxins in poultry Inessa B. Izmailovich1, Marcin W. Lis2 Department of Animal Nutrition, Belarusian State Agricultural University in Horky, Belarus Department of Veterinary, Animal Reproduction and Welfare, University of Agriculture in Krakow, Poland 1 2 e-mail: insera@tut.by Mycotoxins are an invisible group of feed components of varying toxicity and the waste products of mold fungi. The first mycotoxin adsorbents are natural minerals. The basis for the second generation of the adsorbents have become organic substances. The third generation of adsorbents consists of two parts: mineral and organic. Representative of such adsorbents is phytomineral “Fungisorb”. We studied the effect of the adsorbent of mycotoxins on the efficiency of growing broilers. Studies have shown that the use of “Fungisorb” facilitated the intensification of growth of broiler chickens while reducing cost of feed per unit of live weight gain. By the end of the experiment the difference in body weight between the control and experimental groups was 88.2 g, which is 4.0% higher than the control. At the same time the overall cost of feed to weight gain in the experimental group were higher by 3.8 kg than in the control, and the cost of feed to live weight gain per 1 kg were below by 3.6% increase in the experimental group, indicating that a higher ratio of nutrients feed the chickens of experimental group. In the blood of chickens of experimental group compared to the control was observed a statistically significant difference in the content of red blood cells (2.57 ± 0.08) 1012/L against (2.28 ± 0.07) in the control, which provided more substantial opportunities for the effective implementation physiological functions and more rapid growth of experienced livestock. In addition the content of leukocytes was (28.3 ± 0.43) 109/L in experience against (26.9 ± 0.31) in the control; hemoglobin (89.4 ± 1.32) g/L in experiment against (87.5 ± 1.22) in the control. Cellular and humoral factors to protect the body from the chickens of experimental group had advantages. Thus, the use in rations of broiler chickens adsorbent “Fungisorb” stimulates metabolism, strengthens the resistance of the body, increases the growth rate of young growth, while reducing the cost of feed to weight gain, which is economically efficient. Studies supported by DS 3263. 141 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Trends in current research on avian influenza viruses Michał Jóźwiak, Krzysztof Wyrostek, Edyta Świętoń, Monika Olszewska-Tomczyk, Krzysztof Śmietanka, Zenon Minta Department of Poultry Diseases, National Veterinary Research Institute, Puławy, Poland e-mail: michal.jozwiak@piwet.pulawy.pl Avian influenza virus (AIV) causes the highest economic losses in poultry amongst all infective agents. Due to their high variability, both highly and low pathogenic AIV are capable of adaptation to new hosts species, including humans. Research on avian influenza viruses are currently carried out in several main directions. The role of wild birds as a reservoir of LPAIV in the pathogenesis of infections in poultry is underlined. Numerous studies focus on determination of adaptability of these viruses, including molecular characterization of the direction of their genetic variability. Next generation sequencing, which enables sequencing of whole viral genomes, plays a key role. Methods of genetic engineering facilitate studies on the significance of mutations and the role of individual genes. The analysis of nucleotide sequences allows to determine pathogenicity, zoonotic potential and virus resistance to antiviral agents. The participation of some compounds of host immune system in inducing virus 142 variability has not been yet excluded. Much attention has been paid to the role of environmental stressors in virus adaptation. The role of coinfections (especially with immunosuppresive viruses) in AIV adaptation is particularly underlined. The mammalian model (eg. mouse BALB/c) has become a promising object in research on viral variability and zoonotic potential of AIV. A further step in this study may be the use of strains of mice with innate immune defect. The trials of evaluation of protective properties of newly constructed vaccines, mostly vector vaccines, are undertaken. The use of viral, bacterial and yeast vectors is described. Great emphasis is placed on the studies of innate immune response in birds, based on the level of expression of proinflammatory cytokines and pattern recognition receptors and on identification of the potential factors inducing this response. Many studies are devoted to the role of TLR receptors in the signaling pathway of immune response. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Selection of the best reference gene in real-time qPCR studies in ovarian tissue of hens treated with silver nanoparticles Dorota Katarzyńska, Małgorzata Grzesiak, Andrzej Sechman Department of Animal Physiology and Endocrinology, University of Agriculture in Krakow, Poland e-mail: katdorota@op.pl To achieve accurate and reproducible expression profiles of selected genes by real-time qPCR it is crucial to use reliable housekeeping genes. It should be validated for each target cell or tissue type to confirm that it remains unaltered at all points of the experiment. The aim of this research was to evaluate the stability of 4 reference genes and to select the best one for normalization of the gene expression analysis in the ovarian tissues of hens treated with colloidal silver nanoparticles (AgNPs). Four genes were selected for the validation: HPRT (hypoxanthine phosphoribosyltransferase), HMBS (hydroxymethylbilane synthase), VIM (vimentin), which were reported to be the most stable in the chicken ovary according to Olias et al. (2014), and 18S rRNA gene as one of the most commonly used reference gene. The expression levels of these four genes were measured in 51 samples collected from ovarian follicles of control hens (n = 17) and hens receiving the 13 nm AgNPs (n = 17) or 50 nm AgNPs (n = 17) for 14 consecutive days. The stability of the reference genes was determined by three different descriptive statistics, i.e. geNorm, NormFinder and BestKeeper, which produced highly comparable results. The HPRT gene was characterized by the lowest Cq variation and was recommended as the most suitable gene by the BestKeeper algorithm. Similar results were obtained using the geNorm and NormFinder. Outputs from these programs demonstrated that HPRT and VIM genes were the most stable. In conclusion, to achieve accurate comparison of gene expression levels, two or more reference genes should be used for data normalization. The combination of HPRT/VIM genes are suggested to be the most suitable for gene expression evaluation in the all chicken ovarian samples. References Olias P., Adam I., Meyer A., Scharff C., Gruber A.D. 2014. Reference genes for quantitive gene expression studies in multiple avian species. PLoS One, 9(6), e99678. Supported by NCN grant no 2014/15/N/NZ9/01435. 143 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Microstructural features of the Harderian gland in Common pheasant (Phasianus colchicus talischensis) and hybrid of Italian Amber and Common pheasant Joanna Klećkowska-Nawrot1, Karolina Goździewska-Harłajczuk1, Karolina Barszcz2, Artur Kowalczyk3, Ewa Łukaszewicz3 1 Department of Animal Physiology and Biostructure, Faculty of Veterinary Medicine, University of Environmental and Life Sciences in Wroclaw, Poland 2 Department of Morphological Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Poland 3 Division of Poultry Breeding, Institute of Animal Breeding, University of Environmental and Life Sciences in Wroclaw, Poland e-mail: lestat_v@poczta.onet.pl The goal of this study was a characterisation of the histological and histochemical morphology of the Harderian gland in the wild and a hybrid of the wild and domestic birds. The staining with haematoxylineosin, methyl green-pyronin Y, periodic acid-Schiff, alcian blue pH 2.5, aldehyde fuchsin and Hale’s dialyzed iron were performed. In both species the Haredrian glands had multilobar tubuloacinar structure type I. The Harderian glands in birds were located in the orbit near the inter-orbital septum, between medial rectus muscle, pyramidal muscle of the third eyelid and ventral oblique muscle. In a common pheasant the gland was wider in the proximal and distal part. The common pheasant had more elongated lobes of the Harderian gland than in the hybrid one. In the common pheasant, the glandular cells were 144 darkly stained – serous secretion and lightly stained – mucous secretion. In the hybrid the glandular cells had seromucous secretion. Central lobular space, interacinar space, and apical parts of the acini of the Harderian glands were filled with many lymphocytes and plasma cells especially in the common pheasant, where all centers of big lobes were abundantly filled with plasma cells. The plasma cells dominated in the common pheasant`s Harderian gland, while in the hybrid a similar quantity of lymphocytes and plasma cells were present. The periodic acid-Schiff positive cells were dominant in the hybrid. The stainings with periodic acid-Schiff, Hale’s dialyzed iron, alcian blue pH 2.5 and aldehyde fuchsin demonstrate an acid, carboxylated mucopolysaccharides, in glandular cells cytoplasm in the examined birds. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Symbiotic regulates incretin (GIP and GLP-1) expression and secretion in chicken duodenum Paweł A. Kołodziejski, Ewa Pruszyńska-Oszmałek, Maciej Sassek, Daniela Chałupka, Tomasz Szwaczkowski, Paweł Maćkowiak, Leszek Nogowski Department of Animal Physiology and Biochemistry, University of Life Sciences in Poznan, Poland e-mail: pawelbigi@o2.pl There is an evidence that synbiotics have positive impact on the gastrointestinal track. Since 2006, when the European Union banned use of antibiotic growth promoters in poultry nutrition, researchers have tried to find alternatives to these compounds. Problems with finding the alternatives to replace antibiotics in poultry nutrition and potential strategies for supporting animal husbandry are some of the most intensively studied and the most widely discussed areas within the husbandry sciences. Natural alternatives to antibiotics for poultry production are prebiotics and probiotics, alone or in combination (synbiotics). In addition, an interesting aspect of the research is the method of administration. In ovo technique there is a new method of supplementation which allows for application synbiotics in early developmental stage. Simultaneously, enabling the modulation of microbiota in gastrointestinal tract immediately after hatching. Incretins are a group of metabolic hormones which include: GIP (gastric inhibitory peptide) and GLP-1 (glucagon-like peptide-1). Incretins slow down the rate of absorption of nutrients into the blood stream by reducing gastric emptying and may directly reduce food intake. We decided to investigate effect of in ovo symbiotic (SYN1 – Lb. salivarius 3154 + Bi2tos, SYN2 Lb. plantarum 3036 + RFO) administration on incretin expression and secretion in chicken broilers. We found a decrease of GLP-1 and GIP expression in chicken duodenum, and the effect of SYN1 was stronger than SYN2. Moreover, we investigated incretin level in blood serum, and we found statistically significant changes in GIP and GLP-1 concentrations between experimental groups. Results of the experiment indicate that synbiotics regulate incretins signaling and secretion in broiler chickens. This project has received funding from European Union’s Seventh Framework Programme for research, technological development demonstration under grant agreement No. 311794 the Ministry of Science and Higher Education No. W171/7.PR/2014. 145 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Optimising behavioural tests for studying cognitive bias in poultry Ľubor Košťál1, Mária Horváth1, Katarína Pichová1, 2 1 Slovak Academy of Sciences, Institute of Animal Biochemistry and Genetics, Ivanka pri Dunaji, Slovakia 2 Faculty of Natural Sciences, Comenius University in Bratislava, Slovakia e-mail: lubor.kostal@savba.sk Although the connection between emotions and cognition is broadly accepted in humans, the potential of this interaction as a source of information about the animal emotions has been discovered only recently. Since the seminal study of Harding et al. (2004) there have been published many studies exploring the cognitive bias in numerous species, including birds and poultry. In our laboratory we are studying this phenomenon for several years. We have used two model animals – Japanese quail and laying hen and several methodological approaches. The most of our studies are based on the operant conditioning techniques. For this purpose we have developed custom operant conditioning chambers for quail and domestic chicken with touch-screen monitor. These devices are driven by the Biopsychology Toolbox, a free, open source Matlab Toolbox for the control of behavioural experiments. There are many factors affecting operant discrimination learning as well as the ambiguous stimuli tests. One of them is the value of the reward. 146 We have used pelleted food (requires food deprivation to keep birds motivated) as well as highly palatable mealworms (does not require food deprivation). To enable testing without food deprivation we have developed automatic mealworm dispenser. We have tested also the other approaches. We attempted to train quail on a go/no-go procedure to discriminate between two visual stimuli (cardboard lids of white and dark grey) associated with outcomes of a different value (palatable and unpalatable mealworms hidden underneath). Nevertheless, this technique that worked in starlings (Bateson and Matheson 2007) did not prove to work in quail. We have also adopted the spatial judgment task of Burman et al. (2008) for the use in quail. Using suitable reward (mealworms) and punishment (white noise) it proved to be applicable in quail (e.g. for studying the effect of mothering). In the presentation we will discuss perspectives of the cognitive bias testing in poultry. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Quality of native breed hen eggs stored under different conditions Józefa Krawczyk Department of Animal Genetic Resources Conservation, The National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: jozefa.krawczyk@izoo.krakow.pl The aim of the study was to determine the quality of eggs from 6 breeds/lines of hens included in the conservation programme after 28 days of storage in different temperature and relative humidity conditions. Tests were conducted with class M eggs (53–63 g) from 36-week-old hens: Greenleg Partridge (Z-11), Yellowleg Partridge (Ż-33), Sussex (S-66), Leghorn (H-22), Rhode Island Red (R-11) and Rhode Island White (A-33). After laying, eggs were divided into three groups with 20 eggs per group. Group I were fresh eggs, group II were refrigerated eggs (average temperature 4–5°C, 29–36% humidity), and group III were cool-stored eggs (average temperature 14–16°C, 40–45% humidity). Interior egg quality and shell quality for eggs from groups II and III were evaluated after 28 days of storage. Eggs from native and local breeds hens were found to show considerable variation in many quality traits. Cool-stored eggs for 28 days were characterized by a significant deterioration in thick albumen quality compared to eggs stored in a refrigerator. Poorest results in this respect were obtained by eggs from Greenleg Partridge hens. These results suggest that eggs from native hen breeds should be refrigerated because when stored at 14– 16°C they lose too much of their weight, while thick albumen quality parameters may downgrade them from marketable class A. 147 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Carcass muscling and colour in poulards from Sussex (S-66) hens Józefa Krawczyk, Joanna Obrzut Department of Animal Genetic Resources Conservation, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: joanna.obrzut@izoo.krakow.pl The aim of the study was to determine the effect of poulardization of Sussex (S-66) hens on body weight and selected meat quality parameters. The experiment used 100 Sussex (S-66) pullets, half of which were poulardized at 10 weeks of age. The control group were unsterilized pullets. Throughout rearing, all birds were fed ad libitum the same diets and kept on litter with outdoor access, under optimal environmental conditions. At 23 weeks of age, the pullets and poulards reached a body weight of 1617 g and 1726 g, respectively. The poulard group was characterized by 148 higher dressing percentage and weight of breast and leg muscles (P ≤ 0.05). Compared to hens, poulards had a greater content of abdominal fat (by 22.8 g), but with large variation in this trait the differences were not statistically confirmed. The groups also differed in carcass colour, which was lighter (L*) and yellower (b*) in poulards compared to hens. The results obtained show that the poulardization of Sussex (S66) pullets had a beneficial effect on increasing body weight and improving carcass meatiness. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The effect of breed and free-range production system on quality of meat from hens of two native breeds Józefa Krawczyk, Michał Puchała Department of Animal Genetic Resources Conservation, National Research Institute of Animal Production, Balice near Krakow e-mail: michal.puchala@izoo.krakow.pl The objective of the study was to determine the effect of breed and free-range production system on quality of meat from hens of two native breeds, Greenleg Partridge (Z-11) and Rhode Island Red (R-11). The experimental material consisted of 120 hens of each breed, which were assigned to two treatment groups differing in the housing system: 60 layers were kept on litter without outdoor access (C) and 60 layers were raised on litter with access to free range (FR). At 56 weeks of age 8 hens were randomly chosen from each group, slaughtered and subjected to slaughter analysis. It was found from the study that carcasses from 56-week-old general-purpose hens were characterized by poor muscle development, considerable fat content, low tenderness, and high water holding capacity. In the meat of hens that completed their first year of egg production, the profile of fatty acids was beneficial from the standpoint of human nutrition. The free-range production system reduced carcass fatness, enhanced carcass and meat yellowness, and increased the proportion of polyunsaturated fatty acids (both n-6 and n-3) in breast and leg muscles while causing no significant changes in the content of saturated fatty acids. The free-range system improved the meat quality traits in Greenleg Partridge hens (Z-11) to a greater extent than in Rhode Island Red hens (R11). The meat of the Z-11 breed was found to contain less saturated and more unsaturated fatty acids compared to the meat of R-11 hens. 149 Wpływ wybiegowego systemu chowu na jakość mięsa kur dwóch rodzimych ras Józefa Krawczyk, Michał Puchała Dział Ochrony Zasobów Genetycznych Zwierząt, Instytut Zootechniki – Państwowy Instytut Badawczy, Balice k. Krakowa e-mail: michal.puchala@izoo.krakow.pl Celem badań było określenie wpływu rasy i wybiegowego systemu chowu na jakość mięsa dwóch wybranych rodzimych ras kur, tj. zielononóżki kuropatwianej (Z-11) i karmazyna (R-11). Badaniami objęto po 120 kur każdej rasy, które przydzielono do dwóch grup różniących się systemem utrzymania: 60 niosek utrzymywano na ściółce bez dostępu do wybiegu, a 60 na ściółce z dostępem do zielonych wybiegów. W wieku 56 tygodni wybrano losowo po 8 kur z każdej grupy, które poddano ubojowi, a następnie ocenie wartości rzeźnej. Stwierdzono, że tuszki badanych 56-tygodniowych kur ogólnoużytkowych charakteryzują się słabym umięśnieniem, znacznym otłuszczeniem, małą kruchością i dużą wodochłonnością. Mięso badanych kur po pierwszym roku użytkowania nieśnego 150 charakteryzowało się korzystną z punktu widzenia dietetyki człowieka strukturą kwasów tłuszczowych. Wybiegowy system chowu kur wpłynął na zmniejszenie otłuszczenia tuszek, zwiększenie wysycenia barwy tuszek i mięsa w kierunku żółci oraz zwiększenie udziału wielonienasyconych kwasów tłuszczowych PUFA zarówno szeregu n-6, jak i n-3 w mięśniach piersiowych i mięśniach nóg, nie powodując istotnych zmian w zawartości nasyconych kwasów tłuszczowych. Stwierdzono większy wpływ wybiegowego systemu chowu na poprawę cech jakości mięsa u kur zielononóżka kuropatwiana (Z-11) niż u karmazynów (R-11). Wykazano, że mięso kur Z-11 zawiera mniej nasyconych, a więcej nienasyconych kwasów tłuszczowych w porównaniu z mięsem kur (R-11). International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Carcass quality of fattening laying-type cockerels Lucie Kupčíková, Martina Lichovníková, Alena Przywarová, Vojtěch Anderle Department of Animal Breeding, Mendel University in Brno, Czech Republik e-mail: kupcikovalucie@seznam.cz Laying-type cockerels in most countries with developed poultry industry are euthanized immediately after hatching and they are regarded as waste, which is very unethical. Fattening of the cockerels could be an alternative to their killing after hatch. Therefore an experiment was conducted. Males of four laying-type DOMINANT hybrids from the company Hatchery Studenec, s.r.o., two hundred of each, were used in the experiment. Cockerels were fattened on litter until 18 weeks of age. At the age 12 and 18 weeks the carcass yield and the carcass quality of 10 cockerels of each hybrid were evaluated. At 12 weeks of age there was no significant difference (P > 0.05) among the hybrids in live body weight, carcass weight, and thigh and breast muscle proportion. The highest carcass yield was found in D 853 hybrid. At 18 weeks of age the highest live body weight and carcass yield was in hybrid D 853. Anyway there was no significant difference in breast yield among the hybrids. The significantly lowest thighs proportion was found in the hybrids D 109 (P < 0.05). 151 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The effect of day length on broiler breeders’ performance at suboptimal females’ weight Martina Lichovnikova, Sarka Volna Faculty of Agri Sciences, Mendel University in Brno, Czech Republic e-mail: lichovmartina@gmail.com The aim of the study was to compare 13 h and 14 h day lengths on broiler breeders’ performance, while the females had suboptimal body weight at the time of mixing males and females (19 weeks). Two houses on the same farm with the same hybrid combination, technologies and with the same age of the flock were used in this experiment. Approximately 5,400 females were housed in each flock. At 19th week of age the average female weight in both flocks was 234 g lower than the recommendation for this hybrid is in this age. Daily feed intake was increased in both flocks to rich the final body weight, however the live body weight was lower till 21st week of age. At the age of 155 days the day length was firstly prolonged from 8 h to 11 h and consequently the day length was pro- 152 longed till day 186. From this age 13 h day length was used in one flock and 14h in the second one. Hatching eggs weight from 28th to 35th weeks of age was significantly lower (P < 0.05) in flock with 14 h day length in comparison with the flock with 13 h day length (61.9 g vs 60.8 g). Between eggs weight from 28th to 57th weeks of age there was no significant difference (P > 0.05). There was no significant (P > 0.05) difference between the flocks in laying intensity, flock uniformity, proportion of double yolk eggs or hatching eggs. The day length 13 h is sufficient for broiler breeders. Longer lay length can have negative effect on hatching eggs weight at the beginning of laying period in flock with suboptimal body weight at time of mixing males and females. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Asymmetry of roosters’ primary and secondary sexual traits Ewa Łukaszewicz, Martyna Lasoń Department of Poultry Breeding, Wrocław University of Environmental and Life Sciences, Poland e-mail: martyna.lason@up.wroc.pl Asymmetry of roosters’ bilateral traits refers to the random deviations from symmetry of their development. It may reflect the males ability to cope with either environmental or social stress. Males with more symmetrical sexual traits tend to benefit from greater reproductive success than males exhibiting high degree of asymmetry. Moreover, females prefer to mate with more symmetric males. The main goal of the study was to determine the degree of fluctuating asymmetry (FA) of primary (testes) and secondary (wattles) sexual traits of four different lines of roosters. Testes and wattles were collected from roosters in semen production at 40 weeks of age from two laying (ISA Brown and Lohmann Brown) and two broiler breeders (Hubbard Flex and Hubbard F15) lines (15 males per line) immediately after decapitation. Testes and wattles length, width and testes thickness were measured to the nearest 0.1 cm using digital callipers. Testes were also weighed on digital scale to the nearest 0.1 g. The degree of relative FA between the left (L) and right (R) trait was calculated according to the following formula: |L – R|/(L + R). These measures did not deviate from normal distributions with a mean value of zero (Shapiro-Wilks test, P > 0.005 in all cases), which indicates that the traits demonstrate FA. The results of the study show that degree of asymmetry of measured bilateral traits varies between 0.02 to 0.09. The only difference in FA measurements between the lines was found in wattle length, it was significantly (P ≤ 0.05) greater for Hubbard Flex than for Hubbard F15 and Lohmann Brown. The other FA measurements did not differ between the lines. Data obtained show that bilateral traits of all four rooster lines was symmetrical, which means that the males did not suffer from any elevated level of stress. Moreover, there was no evident differences in the degree of FA between meat-type and laying lines of roosters. 153 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Comparison of embryo development at the oviposition of three goose breeds Ewa Łukaszewicz1, Martyna Lasoń1, Murray Bakst1, Krzysztof Andres2, Joanna Rosenberger1, Artur Kowalczyk1 1 Department of Poultry Breeding, Wrocław University of Environmental and Life Sciences, Poland 2 Department of Swine and Small Animal Breeding, University of Agriculture in Krakow, Poland e-mail: martyna.lason@up.wroc.pl Geese are characterized by low reproductive traits and show many characteristics of wild birds, i.e. short, seasonal reproductive period, low number of laid eggs, fertility and hatchability rarely exceeding 80%, as well as high embryo mortality. All these factors result in low number of goslings (about 30) obtained from one goose per year. Taking the above and a large variety of goose breeds into consideration, the optimisation of egg handling and incubation conditions, adapted even to breed specific requirements, become extremely important. These might be achieved by getting deep knowledge about early embryo development. The main goal of project was to determine whether goose genotype affects embryo development i.e., if embryos from different breeds are in the same stage of development at the oviposition. Freshly laid eggs from three breeds: White Koluda, Bilgoraj, Zatorska, were storage at 14–16°C before examination. After distinguish the egg status (fertilized 154 or not), blastoderm was isolated and cleaned off the egg-white and yolk, and then analysed under the stereo microscope and staged according to Eyal-Giladi and Kochav (EGK, Roman numerals) and Hamburger and Hamilton (HH, Arabic numerals) procedures. From each breed at least twenty blastoderms were staged. There was a wide range in embryo stages despite the breed. Blastoderms between Stage X EGK to 2 HH appeared in all breeds. White Koluda embryos were the least developed, majority of them did not exceed Stage X EGK. Most of Bilgoraj and Zatorska goose embryos were at Stage X EGK or XIII EGK, which is characterized by complete hypoblast formation. The results showed no evident differences in embryo development of examined goose breeds however, since the other studies indicate that stage of embryo development at the oviposition determine it further survival during egg storage, they provide information allowing to improve the effectiveness of goose breeding. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The use of the quail chorioallantoic membrane as an experimental in vivo model Mariana Máčajová1, Monika Buríková1, 2, Ivan Čavarga1, 3, Pavel Výboh1, Boris Bilčík1 1 Institute of Animal Biochemistry and Genetics, Slovak Academy of Sciences, Bratislava, Slovakia 2 Cancer Research Institute BMC, Slovak Academy of Sciences, Bratislava, Slovakia 3 St Elizabeth Oncological Institute, Bratislava, Slovakia e-mail: mariana.macajova@savba.sk An excellent alternative to the use of experimental animals for in vivo testing (e.g. toxicity, potential drugs and biomaterials testing, cancer diagnosis and treatment) is avian chorioallantoic membrane (CAM). CAM is an extraembryonic membrane which forms a primary respiratory and excretion organ of the embryo. It is thin and transparent, with dense capillary network. CAM is structurally similar to the retina, buccal mucosa, lungs, placenta and blood brain barrier tissues and offers multiple advantages over other in vivo models. In addition to the low cost, simplicity, and possibility to continuously visualize the test site, CAM model fulfils the 3Rs concept. Most frequently used is chicken CAM, however, Japanese quail CAM can be used with similar results. In our experimental work we used ex ovo technique with quail embryo. Tested substances can be applied systematically or topically on CAM surface. We examined angiogenic effect of several compounds (e.g. endogenous peptide leptin, ghrelin, heparin) admin- istered in ovo or on ex ovo culture. Changes in blood vessel density were quantified by the fractal analysis. Results clearly showed stimulating effect of leptin on blood vessel development. In another series of experiments we explored possible use of quail CAM for photodynamic diagnosis and therapy of cancer. Photodynamically active drug Hypericin (HYP) applied on tumour implanted on CAM surface in fluorescent light better visualized location of tumour, compared to white light. Addition of low density lipoproteins as a delivery molecule improved even more detectability of tumour position. Our results indicate that Japanese quail CAM model is useful tool for the study of antivascular therapy and tumor angiogenesis, development of new biophotonic techniques as well as novel drug testing. This research was supported by VEGA grant No. 2/0102/15. 155 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Activity of antioxidant enzymes and degree of lipid peroxidation in eggs from Greenleg Partridge hens depending on the housing system Renata Muchacka1, Ewa Sosnówka-Czajka2, Iwona Skomorucha2 Department of Animal Physiology and Toxicology, Institute of Biology, Pedagogical University of Cracow, Poland 2 Department of Technology, Ecology and Economics of Animal Production, National Research Institute of Animal Production, Balice near Krakow, Poland 1 e-mail: rmuchacka@up.krakow.pl The aim of the study was to determine the activity of antioxidant enzymes and the level of reduced glutathione (GSH) and malondialdehyde (MDA) in eggs of hens kept in three housing systems. The experiment used hens of the Greenleg Partridge conserved breed. Birds were fed ad libitum standard diets based on layer concentrates. Throughout the experiment, birds had free access to water. At 18 weeks of age, hens were assigned to experimental groups differing in the housing system: group I – furnished cages (750 cm2/bird), group II – litter (9 birds/m2), group III – litter (9 birds/m2) with outdoor access (2.5 m2/bird). At 44 weeks of age, 36 eggs (6 eggs from each group) were collected to determine the yolk and albumen concentrations of MDA and GSH, and the ac- 156 tivity of antioxidant enzymes: superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT). The results were statistically analysed by oneway analysis of variance and significant differences were estimated with Duncan’s multiple range test. The activity of SOD and GPx in yolk and albumen was significantly higher for hens kept on litter (groups II and III) compared to caged birds (group I). The highest GSH concentration was observed in yolk and albumen of eggs from hens in group III, and the lowest MDA concentration was found in eggs of hens from groups II and III. These results suggest that housing system can influence the antioxidant status, and thus the quality of eggs from Greenleg Partridge hens. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effects of herbal mixture and short-term aflatoxin exposure on performance, some blood lipid peroxide and glutathione redox parameters of laying hens Mangesh Nakade, Krisztián Balogh, Erika Zándoki, Zsolt Ancsin, Andrea Bócsai, Márta Erdélyi, Miklós Mézes Department of Nutrition, Faculty of Agricultural and Environmental Sciences, Szent István University, Hungary e-mail: drmangesh23@gmail.com Cereals, the most commonly used feedstuffs are often contaminated with mycotoxins, secondary metabolites of moulds. Aflatoxin (AF) is a hepatotoxic mycotoxin, and, as others, it is thought to provoke oxygen free radical formation and induce oxidative damage in livestock. A body has some protection against reactive oxygen metabolites in form of antioxidant defense system. Many of herbs are capable of scavenging free radicals, thus phytobiotics might be useful antioxidant additives in poultry nutrition. The purpose of the present study was to investigate the short-term effect of AF (0.17 mg/kg) on lipid peroxidation and some glutathione redox parameters in layers, and to evaluate the efficacy of an herbal mixture (Herbamix® Basic Premix 600 mg/kg) for counteracting short-term (36-hours) aflatoxicosis. Thus 60 Bovans Goldline hens were divided in four groups (Control, Aflatoxin, Herbamix, Aflatoxin + Herbamix) and 5 birds of each group were slaughtered and sampled at 12th, 24th, 36th hours of the experiment. Parameters of the lipid peroxidation (malondialdehyde, MDA) and the glutathione redox system (reduced glutathione (GSH) content and glutathione peroxidase (GPx) activity) were measured in blood plasma and red blood cell (RBC) haemolysate. The results revealed that AF did not have an effect on feed intake, body and liver weight, but MDA content was increased, in blood plasma and RBC haemolysate, as the effect of the combined treatment at 36 hours of the trial. GSH content was significantly reduced due to the Herbamix treatment in RBC haemolysate, but no other changes were found. The activity of GPx was lower in all treated groups in blood plasma, and the difference was significant at 36 hours as compared with the control, while no change was found in RBC haemolysate. The results are contradictory with previous findings, probably due to the short-term exposure, and a moderate modifying effect of Herbamix on AF toxicity. 157 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of age of broiler chickens on the expression level of genes coding for calpain (CAPN1, CAPN2, CAPN3) and calpastatin (CAST) in breast muscles Joanna Nowak1, Katarzyna Połtowicz1, Katarzyna Piórkowska2 Department of Animal Genetics and Breeding, National Research Institute of Animal Production, Balice near Krakow, Poland 2 Department of Animal Genomics and Molecular Biology, National Research Institute of Animal Production, Balice near Krakow, Poland 1 e-mail: joanna.nowak@izoo.krakow.pl The objective of the study was to determine the effect of broiler chicken age on the expression level of genes coding for calpain 1 (CAPN1), calpain 2 (CAPN2), calpain 3 (CAPN3) and calpastatin (CAST) in breast muscles. Ross 308 cockerels were studied. At 35 (group I) and 42 days of age (group II), 20 chickens whose body weight was similar to average weight at a given age, were selected from each group for slaughter. Within 15 min postmortem, samples of pectoralis superficialis muscles were collected from chickens to isolate RNA and to determine the expression level of selected genes. After 24-hour chilling of carcasses at 4°C, chickens were subjected to slaughter analysis. On this basis, the percentage of breast muscles in relation to carcass weight with giblets was calculated. The expression data were analysed using Sequence Detection System Software v. 2.0 (Applied Biosystems). The results were statistically analysed with Statistica 10 PL. 158 The final body weight of 42-day-old chickens (2657 g) was higher by 864 g on average (P ≤ 0.01). The dissection results show that significantly better breast muscling was characteristic of birds subjected to a 7 days longer rearing period (P ≤ 0.01). A higher transcript level of the examined genes was characteristic of breast muscles from younger chickens. However, no statistically significant differences in the expression of CAST, CAPN1 and CAPN2 genes in breast muscles from birds of different age groups were found. Significantly stronger expression of the CAPN3 gene was observed in breast muscles from younger birds (group I; P ≤ 0.05). Analysis of the correlations showed that expression of the CAPN3 gene was significantly negatively related to body weight (–0.383; p = 0.018) and percentage of breast muscles (–0.468; p = 0.003). Our study demonstrated that the age of chickens has an effect on expression of the CAPN3 gene in breast muscles. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Expression of aromatase in the testis, epididymis and ductus deferens of the turkey (Meleagris gallopavo) with yellow semen syndrome Laura Pardyak1, Alicja Kamińska1, Ewelina Górowska-Wójtowicz1, Anna Hejmej1, Mariola Słowińska2, Małgorzata Kotula-Balak1, Andrzej Ciereszko2, Barbara Bilińska1 1 Department of Endocrinology, Institute of Zoology, Jagiellonian University in Kraków, Poland Department of Molecular Andrology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Olsztyn, Poland 2 e-mail: ewelina.gorowska@uj.edu.pl Yellow semen syndrome (YSS) is endemic within the population of domestic turkeys. No prior information concerning the YSS is available. Most of the existing evidence points to a viral origin, although other causes of the YSS appearance are not excluded. Yellow semen, when used for insemination, is characterized by lower quality and reduced fertility. Since an essential role of estrogens in male fertility has been reported in mammals and several lower vertebrate species, we aimed to examine the expression of aromatase in the testis, epididymis and ductus deferens of YSS turkeys compared with the turkeys producing white semen (WS). The study was performed on tissue samples collected from six adult YSS turkeys and six WS. Morphological analysis of testicular, epididymal and ductal deferens cells was performed by staining with hematoxylin-eosin, whereas localization of aromatase and its expression were analyzed by immunohistochemistry and Western blot, respectively. In testes of both YSS and WS turkeys, fully differentiated germ cells, reflecting normal spermatogenesis were observed. No obvious alterations were found in the epididymis and ductus deferens morphology of YSS turkey compared to WS individuals. Immunohistochemical analysis revealed aromatase as localized to Leydig cells and germ cells residing in the basal compartment of the seminiferous tubules. Immunopositivity was detected in the cell cytoplasm, whereas nuclei were negative. Intensity of the staining for aromatase increased along the reproductive tissues being weak to moderate in testicular cells, strong in epididymal cells, and very strong in epithelial cells and smooth muscle cells of the ductus deferens. Moreover, statistically significant increase in aromatase expression at protein level (P < 0.001) was detected in all the YSS tissues compared to those of WS turkeys. Altogether, the results indicate the potential relationship between estrogens and appearance of the YSS. Supported by K/ZDS/005402. 159 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The influence of electromagnetic field emitted by mobile telephony on chick hematology parameters Krzysztof Pawlak, Bartosz Bojarski, Marcin Lis, Weronika Kanik, Jerzy Niedziółka, Barbara Tombarkiewicz Department of Veterinary Science, Animal Reproduction and Animal Welfare, Institute of Veterinary Sciences, University of Agriculture in Krakow, Poland e-mail: rzpawlak@cyfronet.pl Artificial electromagnetic fields (EMF), which are a consequence of human progress, begin to play a considerable role in shaping the Earth’s electromagnetic environment. Among these, the fields generated by mobile telephony deserve special attention due to their prevalence. There are no data in the literature concerning the impact of EMF on the hematological picture of animal organisms in the embryo stage. Therefore, this study attempted to determine the effects of a 1800 MHz electromagnetic field on chick hematology parameters. There were three hundred sixty eggs used in the experiment. They were kept in incubators. The eggs incubated using standard procedures were divided into 2 equinumerous groups: group I (control) and group II (experimental). The experiment was conducted in three sequential test series. In each series, embryos from experimental incubators were exposed daily to electromagnetic field from the first to the last day of incubation. EMF exposure time was as follows: Series I – 26 min per day (13 times per 2 minutes), Series II – 40 min per day 160 (10 times per 4 minutes), and Series III – one 40-minute emission. Blood samples were collected from newly hatched chicks. The samples were obtained from 10 exposed and 10 control individuals. The total number of erythrocytes (RBC), the total number of white blood cells (WBC), hematocrit (Hct) value, and the total hemoglobin concentration (Hb) were determined. The obtained results showed the influence of the EMF on the image of the chicks blood. The significant increase of the WBC was observed as well as the value of Hct and Hb concentration were increased in the animals exposed to the EMF for 26 minutes a day. Whereas in animals that were exposed to the EMF for 40 minutes per day not only the mentioned parameters were increased, but also the RBC and MCHC. No significant changes were noted in MCV and MCH values. This research was financially supported by the statutory activity: DS 3263. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Combined effect of T-2 toxin and deoxynivalenol on regulation of the glutathione redox system in broiler chicken Csilla Pelyhe1, Benjámin Kövesi1, Erika Zándoki2, Miklós Mézes1, Krisztián Balogh1, Balázs Kovács1 Faculty of Agricultural and Environmental Sciences, Szent István University, Hungary Mycotoxins in the Food Chain Research Group, Hungarian Academy of Sciences – Kaposvár University, Hungary 1 2 e-mail: pelyhe.csilla@gmail.com The aim of this study was to evaluate the combined effect of T-2 toxin and deoxynivalenol (DON) in 3-week-old broiler chickens, at the doses of A: 0.25 mg T-2 toxin and 5.0 mg DON/kg feed, B: 1.25 mg T-2 toxin and 12.5 mg DON/kg feed and C: 2.5 mg T-2 toxin and 25 mg DON/kg feed for 14-days. The changes in gene expression of the GPx4 (phospholipid hydroperoxide glutathione peroxidase), GSS (glutathione synthetase) and GR (glutathione reductase) were measured. Liver samples were taken on days 1, 2, 3, 7 and 14 after the beginning of a feeding trial. Changes of the gene expression of target genes and endogenous control gene, GAPDH (glyceraldehyde 3-phosphate dehydrogenase) were determined in the liver, using qPCR with TaqMan probes. GPx4 is described in avian and fish species to be far more important in the antioxidant defense system as compared to mammals; elevated on day 1, but showed only minor changes later. GSH plays important role in the antioxidant system, protecting cells from oxidative stress. GSS, which is the final enzyme in the glutathione (GSH) biosynthesis pathway, showed an inhibition and a slow, but continuous, elevation in gene expression, which exceeded the control levels after 14-days. During the observed period GR, which catalyzes the reduction of glutathione disulfide (GSSG) to GSH, repairing its antioxidant abilities, showed decreased levels which later elevated and exceeded control levels, but decreased on day 14 again. The members of glutathione-redox system responded to the mycotoxin exposure in gene expression levels, which can be in association with emerging formation of reactive oxygen species (ROS). The transcription of antioxidant enzymes are regulated by Nrf2-ARE pathway, which reacts sensitively to oxidative stress and modulates the transcript of the antioxidant defense system. This study was supported by the János Bolyai Research Scholarship (BO/261/13) of the Hungarian Academy of Sciences and Hungarian National Research fund (OTKA PD-104823). 161 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Thermographic evaluation of feather cover damage in laying hens as a tool for welfare assessment Katarína Pichová1, 2, Boris Bilčik1, Ľubor Košťál1 1 Institute of Animal Biochemistry and Genetics, Slovak Academy of Sciences, Ivanka pri Dunaji, Slovakia 2 Faculty of Natural Sciences, Comenius University in Bratislava, Slovakia e-mail: katarina.pichova@savba.sk The quality of the feather cover represents established poultry welfare indicator because it can reflect the health as well as outcome of animal-housing and animal-animal interactions. The infrared thermography for its noninvasiveness and repeatability is considered as a potential alternative to commonly used visual feather scoring. The aim of this study was to measure feather cover damage of laying hens kept in two housing systems and to compare the outcome of the infrared thermography evaluation with the standardized feather scoring system based on the Welfare Quality® poultry protocol. Thirty beak trimmed Dekalb White laying hens were starting from 16 weeks of age housed in litter pens (n = 15) or enriched cages (n = 15). During the next four months the feather damage was assessed in two week intervals and then once again at week 38 of the experiment. The assessment was carried out on four body regions: head and neck, back and rump, belly, and underneck and breast using feather scoring and two parameters derived from 162 the infrared thermographic measurement (the difference between the body surface temperature and the ambient temperature and the proportion of featherless areas, represented as areas with the temperature higher than 33.5°C). Data were analysed using the generalized linear mixed model with housing, body region and time as the fixed effects. The effects of housing, time, region and housing x time interactions on feather damage assessed by the feather scoring and by both infrared thermography measures were all significant (P < 0.001). Feather scoring as well as the thermography parameters revealed higher damage in enriched cages compared to deep litter pens with belly and back and rump being the most affected body regions. There was significant positive correlation between the thermographic parameters and the feather scoring (P < 0.001). These findings support the feasibility of infrared thermography for the assessment of the feather cover quality in laying hens. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Applications of the CHD locus in sex identification in birds Angelika Podbielska, Anna Radko, Agnieszka Szumiec Department of Animal Genomics and Molecular Biology, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: angelika.podbielska@izoo.krakow To identify the sex of birds, techniques of molecular biology are used increasingly, because nearly 50 percent of birds are sexually monomorphic. The CHD gene has been widely used to determine the sex of birds, due to the high degree of conservatism and the presence of polymorphic variants of the Z and W chromosomes. PCR products are usually visualized on agarose gel, where PCR amplification of the CHD gene produces a double (ZW) and single (ZZ) bands in females and males, respectively. In our laboratory, we have implemented a technology for identifications the bands using capillary electrophoresis. 69 individuals were tested from genera Columba and 45 individuals from genera Aprosmictus, Cacatua, Myiopsitta, Platycercus, Polytelis and Psittacula. Genomic DNAs were extracted from feathers and buccal swabs. DNA isolates were amplified by PCR for the CHD marker with primers, which was labelled with fluorescent dye: 6-FAM. Sex-specific amplification of CHD gene was amplified using the QIAGEN Multiplex PCR Kit, the amplified products were separated on 3100xl Genetic Analyzer and genotyped using GeneMapper software (Applied Biosystems). PCR products were obtained for all samples. Variation in the intronic sizes of CHD-W and CHD-Z gene was detected by capillary electrophoresis. Two different picks of PCR products was produced from CHDW and CHD-Z genes in females and one pick of PCR products from CHD-Z gene in males. The tested material had a high genetic diversity, were identified an evident difference in products size. Additionally 5 gene variants in the range of 268–320 bp were observed. 163 Wykorzystanie konserwatyzmu genetycznego sekwencji mikrosatelitarnej genu CHD do identyfikacji płci u ptaków Angelika Podbielska, Anna Radko, Agnieszka Szumiec Dział Genomiki i Biologii Molekularnej Zwierząt, Instytut Zootechniki – Państwowy Instytut Badawczy, Balice k. Krakowa e-mail: angelika.podbielska@izoo.krakow.pl W związku z faktem, że blisko 50% ptaków należy do gatunków monomorficznych, do identyfikacji płci coraz częściej wykorzystuje się techniki biologii molekularnej. Z uwagi na wysoki stopień konserwatyzmu i występowanie polimorficznych wariantów na chromosomach Z i W, gen CHD znalazł szerokie zastosowanie do oznaczania płci u ptaków. Do jego analizy stosuje się techniki elektroforetyczne oparte na wizualizacji produktu PCR na żelu agarozowym. Po uzyskaniu obrazu w postaci dwóch lub jednego prążka ustala się odpowiednio żeńską lub męską płeć badanego osobnika. Obecnie w Laboratorium Genetyki Molekularnej IZ PIB wdraża się metodę identyfikacji płci z zastosowaniem techniki elektroforezy kapilarnej. Dotychczas przebadano 78 osobników z rodzaju Columba i 45 osobników z rodzajów Aprosmictus, Cacatua, Myiopsitta, Platycercus, Polytelis i Psittacula. Materiał stanowił genomowy DNA uzyskany z piór 164 i wymazów. Reakcję PCR przeprowadzono z zastosowaniem wyznakowanej fluorescencyjnie barwnikiem 5-FAM sekwencji mikrosatelitarnej genu CHD. Produkty PCR poddano analizie fragmentów długości w sekwenatorze 3100xl Genetic Analyzer, a wyniki w postaci pików określonych liczbą par zasad odczytano w programie GeneMapper® Software 4.0. Dla wszystkich osobników otrzymano produkty PCR. Dla badanych samic uzyskano 2 produkty PCR różnej długości z chromosomów CHD-W i CHD-Z, natomiast dla samców dwa produkty tej samej długości z chromosomów CHD-Z. Pozwoliło to na obserwację w obrazie elektroforetycznym jednego lub dwóch pików, co umożliwiło ustalenie płci. Uzyskano duże zróżnicowanie długości fragmentów DNA świadczące o dużym zróżnicowaniu genetycznym badanego materiału. Otrzymano 5 wariantów genu w zakresie 268–320pz. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of early feed restriction on liveability, rearing performance and meat quality of broiler chickens Katarzyna Połtowicz, Joanna Nowak Department of Animal Genetics and Breeding, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: katarzyna.poltowicz@izoo.krakow.pl The aim of the study was to determine the effect of feed restriction during the early period of rearing broiler chickens on production results and meat quality. In total, 540 sexed Ross 308 chicks (cockerels) were randomly assigned to two groups with 270 birds per group and reared on litter until day 42 of age under optimal environmental conditions. All birds were fed the same complete starter, grower and finisher diets containing 22, 20.5 and 20.5% CP and 2990, 3130 and 3130 ME/kg, respectively, and had free access to drinking water. Throughout rearing, chickens from group I (control) were fed ad libitum, whereas birds from group II (experimental) received about 66% of the control feed intake at 2 and 3 weeks of age. Throughout the study, individual body weight, feed consumption, and number of dead birds were determined every week. At the end of the rearing period, 30 chickens with average body weight were selected from each group to estimate dressing percentage and to determine carcass and meat quality. The results were statistically analysed with analysis of variance and Duncan’s test. The body weight of chickens from groups I and II averaged 2806 g and 2527 g, respectively (P ≤ 0.01). The birds from both groups that differed in body weight also showed differences in carcass fatness, but a greater proportion of abdominal fat was observed in group II. There were no differences in dressing percentage and carcass percentage of breast and leg muscles. Restricting body weight gain through feed restriction did not improve the physicochemical characteristics of broiler meat and did not change the incidence of muscles characterized by higher drip loss. This course of action, however, reduced mortality and improved feed conversion (kg feed/kg gain). 165 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effectiveness and safety of the use of antibiotics in fowl Andrzej Posyniak, Krzysztof Niemczuk Department of Pharmacology and Toxicology, The National Veterinary Research Institute, Puławy, Poland e-mail: aposyn@piwet.pulawy.pl Intensification of farming, changing living conditions and effective technologies to gain body weight can lead to new, previously unknown health problems. Changing the structure of the herd, crowding a limited space can promote the rapid spread of bacterial infections. Therefore, before veterinary doctors it is a unique challenge, because bred varieties can vary quite significantly physiology and biochemical course. Therefore, at the stage of accession to the treatment it plays an important role knowledge, which allows you to select the drug and to determine the dosage. For use allowed a significant number of antibiotics, which have a different mechanism of action and scope. The doctor is able to find an appropriate antibiotic for the treatment of bacterial infection, and the use of antibiotics accordance with the instructions may lead to growing antibiotic resistance and the lack of therapeutic success in treating successive flocks of birds. 166 Furthermore, in the wrong, too high a dosage may lead to a direct threat to animal health, the negative impact on the body of the dispensing doctor or operating personnel. Another no less serious threat may be residues of antibiotics, which can negatively affect the bacterial flora of the human gastrointestinal tract. With prolonged exposure, they can modify the composition of the bacterial flora and increase drug resistance. In addition to the above mentioned side effects of massive use of antibiotics on farms is environmental pollution are eliminated active substances and/or products of their metabolism, which after getting into the soil, go into the water and become a secondary source of danger to humans and other living organisms. This problem is all the more important as drugs, depending on the chemical, physical and environmental conditions may be stable and its biological activity for several months or even several years. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 In ovo synbiotic administration affects on metabolic and hormonal profile in broiler chickens Ewa Pruszyńska-Oszmałek, Paweł A. Kołodziejski, Maciej Sassek, Daniela Chałupka, Tomasz Szwaczkowski, Paweł Maćkowiak, Leszek Nogowski Department of Animal Physiology and Biochemistry, University of Life Sciences in Poznan, Poland e-mail: ewaprusz@up.poznan.pl There is evidence that synbiotics have positive impact on the gastrointestinal microflora and they have antibacterial, immunostimulatory, and anti-inflammatory properties. However, the effect of bioactives on hormonal profile and lipid-carbohydrate metabolism is not entirely clear. In the present study, synbiotics were given using technology in ovo. This is a new, very promising technology, consisting in the administration of the bioacitve compounds, directly inside the eggs during their incubation. The aim of the present experiment was to evaluate the effects of in ovo synbiotics administration on hormonal profile and lipids and glucose levels in serum blood of broiler chickens. On d 12 of incubation, the eggs were divided into control group and two experimental groups (SYN1 and SYN2) treated with different synbiotics, administered in ovo. The SYN1 group was injected with a solution containing Lb. salivarius 3154 + Bi2tos, and SYN2 group received Lb. plantarum 3036 + RFO. On days 1, 3, 7, 14, 21, and 42, cockerels were slaughtered and blood was collected. The concentration of insulin, glucagon, leptin and corticosteron was determined in serum blood. Furthermore, we measured blood lipids and glucose levels. We found a rise of glucagon and leptin levels in SYN1 group compared to control broiler chicken, respectively, at 21 and 7 days of age the birds. The effect of synbiotics on corticosteron concentration was noticed only at the beginning of the experiment (on day 4). The cholesterol, triglycerides and glucose concentrations determined in this study were found to be similar among control and experimental groups, whereas free fatty acids level significantly differ between the groups, however, only in the oldest birds. This project has received funding from European Union’s Seventh Framework Programme for research, technological development demonstration under grant agreement No. 311794 the Ministry of Science and Higher Education No. W171/7.PR/2014. 167 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Application of 10 microsatellites DNA markers for parentage control in pigeons Anna Radko1, Agnieszka Szumiec1, Angelika Podbielska1, Tomasz Borkowski2 1 Department of Animal Genomics and Molecular Biology, National Research Institute of Animal Production, Balice near Krakow, Poland 2 Veterinary Laboratory, Coba Diagnostic, Sośnicowice, Poland e-mail: anna.radko@izoo.krakow.pl Microsatellite DNA became an international standard for individual identification and parentage verification of animals. We tested the polymorphism of 10 microsatellite markers and their usefulness for parentage verification in pigeons. Samples were collected from 69 individuals of Polish, German and Belgian breeds from different locations in Poland. The assay involved the following 10 loci: CliµD01, CliµD16, CliµD32, CliµT13, CliµT17, PG2, PG3, PG5, PG6, PG7 and the bird sex marker CHD. We used genomic DNA extracted from feathers and buccal swab samples. DNA extracts were amplified by PCR for the all microsatellite and CHD marker, together in one multiplex reaction. Each of the forward primers was labeled with fluorescent dye: 6-Fam, Vic, Ned and Pet. Markers were amplified using the QIAGEN Multiplex PCR Kit. The amplified products were separated on 3100xl Genetic Analyzer and genotyped using GeneMapper software (Applied Biosystems). 168 In the study we detected 60 alleles, which number per locus ranged from 2 (PG5) to 8 (CliµD01 and CliµT17). Based on the frequency of identified alleles, expected heterozygosity (He) and observed (Ho) were determined as Ho = 0.88 and He = 0.78, respectively. The average negative inbreeding coefficients was Fis = –0.03. The lowest polymorphism in the present study was noted at the PG5 (PIC = 0.34 and H = 0.47), where two alleles identified, and PG6 (PIC = 0.43 and H = 0.59) for which two of three alleles occurred with higher frequency in excess of 94%. The average power of discrimination (PD) was 0.83. The combined power of discrimination values for all 10 loci reached the high value as 0.9999999. The cumulative probabilities of parentage exclusion, when one parent is known, and when two parents are known (PEc1 and PEc2) were 0.9726 and 0.9982, respectively. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Nesting behavior of Western capercaillie (Tetrao urogallus) females in the breeding aviary Joanna Rosenberger, Artur Kowalczyk, Ewa Łukaszewicz, Zenon Rzońca Department of Poultry, Wrocław University of Environmental and Life Sciences, Poland e-mail: joanna.rosenberger@up.wroc.pl Capercaillie (Tetrao urogallus) is forest grouse that need active protection, including releasing captive birds from breeding centers into wild populations. Despite years of research, biology of this species is still poorly understood. Eggs incubation conditions is an aspect which require revision, because the results of artificial breeding (in incubators) are not satisfactory. Understanding the natural breeding process can help to increase hatchability. Out of the ten females selected for observation, eight started the incubation. The studies have focused mainly on length and frequency of the recesses (corresponding to egg cooling) and the egg turning frequency. It has been observed that birds left the nest more frequently between 15:00 and 19:00, however there were not significant differences (p = 0.106). Individual factor did not affect on the preferred hours of recesses (p = 0.207). The total length of the recesses did not depend on the individual preferences (p = 0.326), but was dependent on incubation day (p < 0.001). On the first day females spent outside the nest more time, what suggest that the instinct of incubation, did not fully activate. Nest leaving frequency was depended on individual (p < 0.001). Bird 6 and 4, left the nests significantly more often than the others. This behavior could be caused by weaker condition of females or individual differences. On the other hand, the number of recesses did not affect the total length of stay out of the nest. Egg turning rate frequency was affected by day of incubation (p < 0.001). Females turned eggs more often during first four and last three days of incubation. An hour affect the frequency of turning (p < 0.001). During the day, the females turned eggs on average 1.0–1.3 times/hour, and less at night, between 20:00 and 2:00. Individual factor had an impact on the turning frequency (p < 0.001). Female number 3 turned eggs most frequently (1.4 turns/ hour.), the lower frequency of turnings was found in female number 7 (0.8 turns/hour). 169 Zachowania lęgowe samic głuszca (Tetrao urogallus) w hodowli wolierowej Joanna Rosenberger, Artur Kowalczyk, Ewa Łukaszewicz, Zenon Rzońca Zakład Hodowli Drobiu, Uniwersytet Przyrodniczy we Wrocławiu e-mail: joanna.rosenberger@up.wroc.pl Głuszec (Tetrao urogallus) jest kurakiem leśnym wymagającym czynnej ochrony, w tym zasilania dziko żyjących populacji osobnikami pochodzącymi z hodowli zamkniętych. Mimo lat badań, biologia tego gatunku wciąż jest niedostatecznie poznana. Jednym z aspektów wymagających rewizji są warunki inkubacji jaj, gdyż wyniki sztucznych lęgów (w inkubatorach) są niezadowalające. Poznanie procesu lęgów naturalnych może przyczynić się do zwiększenia wylęgowości piskląt. Spośród dziesięciu samic wytypowanych do obserwacji osiem przystąpiło do inkubacji zakończonej wylęgiem. W badaniach skupiono się głównie na długości i częstotliwości przerw (odpowiadających chłodzeniu jaj) oraz częstotliwości obracania jaj. Zaobserwowano, że ptaki nieco częściej opuszczały gniazdo pomiędzy godziną 15:00 a 19:00, nie były to jednak istotne różnice (p = 0,106). Czynnik osobniczy nie wpływał na preferowane godziny przerw (p = 0,207). Łączna długość przerw nie zależała od indywidualnych preferencji osobnika 170 (p = 0,326), ale była uzależniona od dnia inkubacji (p < 0,001). Pierwszego dnia samice przebywały poza gniazdem dłużej, co sugeruje, że instynkt wysiadywania dopiero się uaktywniał. Częstotliwość opuszczania gniazda była uzależniona osobniczo (p < 0,001). Samice 6 i 4 istotnie częściej opuszczały gniazdo niż pozostałe. Zachowanie takie mogło wynikać ze słabszej kondycji dwóch głuszek lub różnic osobniczych. Z drugiej strony liczba przerw nie wpłynęła na łączną długość przebywania poza gniazdem. Na częstotliwość obrotów jaj wpływał dzień inkubacji (p < 0,001). Samice częściej obracały jaja podczas pierwszych czterech oraz ostatnich trzech dni inkubacji. Godzina miała wpływ na częstotliwość obracania jaj (p < 0,001). W ciągu dnia samice obracały jaja średnio 1,0–1,3 razy/godz., natomiast w godzinach nocnych, pomiędzy 20:00 a 02:00, rzadziej. Czynnik osobniczy miał wpływ na częstotliwość obracania jaj (p < 0,001). Najczęściej jaja obracała samica 3 (1,4 obrotów/godz.), najrzadziej samica 7 (0,8 obrotów/godz.). International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 In ovo synbiotic administration modulates trypsin, lipase and amylase activity in broiler chicken Maciej Sassek, Ewa Pruszyńska-Oszmałek, Paweł A. Kołodziejski, Daniela Chałupka, Tomasz Szwaczkowski, Leszek Nogowski, Paweł Maćkowiak Department of Animal Physiology and Biochemistry, University of Life Sciences in Poznan, Poland e-mail: macsass@up.poznan.pl Probiotics, prebiotics and their combination – synbiotics are a relatively new alternative to antibiotics which are banned in European Unions in poultry nutrition. Synbiotics affect the host’s welfare by improving its survival, growth and health by activating the metabolism and modulating microflora of the gastrointestinal tract. Synbiotic selection and administration are some of the most intensively studied in medical and also husbandry sciences. In ovo technique symbiotic injection is a solution which allows for application of synbiotics in the early developmental stage and modulation of gastrointestinal tract microbiota immediately after the hatching. In this study we decided to investigate effect of symbiotic on enzyme (amylase, lipase and trypsin) activity in pancreas and duodenum content. On the 12th day of incubation, the eggs were randomly divided into three experimental groups and treated in ovo with different solutions. Control group (n = 10) was injected with physiological saline. SYN1 experimental groups (n = 10) were injected with a solution containing Lb. salivarius 3154 + Bi2tos and SYN2 experimental group (n = 10) received Lb. plantarum 3036 + RFO (raffinose family oligosaccharides). On the days 1, 3, 7, 14, 21, and 42, 10 cockerels were sacrificed by cervical dislocation. The activity of pancreatic enzymes was determined based on the collected material. For investigation of amylase, lipase and trypsin activities the colorimetric assay kits were used. We detected some changes in enzyme activity, even on pancreatic level (enzyme activity were measured in pancreas homogenates). However, more visible changes were observed in chicken duodenum content. Such results suggest that in ovo administration of synbiotics can have a long lasting effect in chicken physiology and husbandry. This project has received funding from European Union’s Seventh Framework Programme for research, technological development demonstration under grant agreement No. 311794 the Ministry of Science and Higher Education No. W171/7.PR/2014. 171 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of adding herb extracts to drinking water on broiler chicken productivity, corticosterone level and heterophil: lymphocyte ratio during heat stress Iwona Skomorucha, Ewa Sosnówka-Czajka Department of Technology, Ecology and Economics of Animal Production, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: iwona.skomorucha@izoo.krakow.pl Ross 308 broiler chickens were assigned to 4 experimental groups. During the study, birds were exposed for 6 days to high daily air temperatures (30°C). In addition, from 21 days of rearing, water drinkers of birds from groups 2, 3 and 4 were supplemented with extracts of herbs: lemon balm (Melissa officinalis L.), sage (Salvia officinalis L.) and common nettle (Urtica dioica L.) at 2 ml/l water, respectively. Broilers were kept in compartments on litter at a stocking density of less than 33 kg/m2 and fed ad libitum standard diet. Body weight, feed conversion and mortality were recorded throughout the study. During days of high air temperature (days 31, 33 and 36), blood was collected from 10 chickens per group to determine corticosterone levels, and blood smears were made to calculate the heterophil to lymphocyte (H:L) ratio. These tests were also performed 3 days after the reduction in air 172 temperature (20.7°C), i.e. on day 39 of rearing. The results were statistically analysed with analysis of variance and significant differences were estimated using Duncan’s test. In the first period of rearing, a statistically significant difference in feed conversion (kg/ kg gain) was noted between groups I, II and III and group IV. Group IV was characterized by the lowest bird mortality. Broiler chickens showed reduced corticosterone levels on days 36 and 39 compared to day 31 of the study. On day 33, birds from group III had a lower H:L ratio compared to groups I (p ≤ 0.01) and II (p ≤ 0.05). In conclusion, supplementing the drinking water with herbs failed to improve body weight and feed conversion in broilers exposed to high air temperatures. Birds receiving the nettle extract in water were characterized by better health compared to the other experimental groups. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of housing systems on some blood parameters of broiler chickens during a period of high temperatures Iwona Skomorucha, Ewa Sosnówka-Czajka Department of Technology, Ecology and Economics of Animal Production, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: iwona.skomorucha@izoo.krakow.pl The experiment was conducted during the summer production cycle. Ross 308 broiler chickens assigned to two experimental groups: I – birds kept on litter without access to pasture, II – birds raised on litter with access to pasture from the first day of rearing. During the second rearing period, when outdoor temperatures were high in the 4th and 6th week of the experiment, blood was collected in 10 birds per group to determine the levels of thyroid hormones, glucose, corticosterone, and hematocrit. These tests were performed between 13:00 and 15:00 on days 24, 27, 36 and 41 of rearing. Similar tests were done on the 3rd day after the decrease in external temperature, i.e. on day 31 of growth. The results were statistically analysed with one-way analysis of variance and the differences were estimated using Duncan’s test. In broilers from group II, T3 decreased and T4 increased with an increase in air temperature (p ≤ 0.01). In our study, broiler chickens reared indoors had higher glucose concentrations on days 27 and 31 as well as higher corticosterone concentration on day 27 of the experiment. Broiler chickens raised with outdoor access were characterized by stable blood glucose levels, while the level of corticosterone decreased significantly during the second period of high external temperatures. In birds confined indoors, blood glucose and corticosterone levels increased during the 4th week of growth, i.e. during the first heat wave, and decreased in the 6th week, i.e. during the second period of increased temperature. In both group I and II, blood hematocrit was highly significantly the highest on day 31 compared to the other test days. In summary, even a small increase in ambient temperature beyond the recommendation did influence the broiler’s body homeostasis regardless of the housing system. Broiler chickens with outdoor access during high external temperatures had a normal thermoregulatory system, as indicated by the decrease in the level of T3 thyroid hormone. 173 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of growth hormone on estrogen receptor and egg shell-specific protein gene expression in the chicken shell gland during pause in laying induced by fasting Joanna Socha, Andrzej Sechman, Maria Mika, Anna Hrabia Department of Animal Physiology and Endocrinology, University of Agriculture in Krakow, Poland e-mail: rzhrabia@cyf-kr.edu.pl The avian oviduct is a side of growth hormone (GH) synthesis and endocrine/paracrine/autocrine actions. Recent studies revealed that GH, beside orchestration of ovarian steroid synthesis, regulates gene expression of some egg specific-proteins in the chicken oviduct during maturation. Since concentration of plasma GH decreases during a pause in egg laying in hens, in the present investigation the effect of GH administrations during the pause in laying on concentration of estradiol (E2) and its receptors and egg shellspecific protein gene expression in the chicken shell gland were investigated. The pause in egg laying was induced by food deprivation for 5 days, followed by feeding every second day and from day 10 hens were fed ad libitum. On day 6, 13 and 17–20 of experiment the shell gland was isolated from hens: (1) control, (2) fasted and (3) fasted and injected with chicken GH. In tissues, concentration of steroid (RIA) and mRNA of E2 receptors (ERalpha, ERbeta), ovocleidin-116, ovocalyxin-32 and -36 (real-time PCR) were examined. 174 The pause in egg laying occurred between the days 6 and 13. Fasting caused: (1) decrease in the tissue concentration of E2 on the day 6 and 13, (2) increase in gene expression of both ERs, and ovocalyxin-36 on the day 6 of the experiment. Administrations of GH (1) slightly abolished the fasting-induced decrease in tissue E2 concentration, (2) elevated the expression of ERalpha and ERbeta on the days 6 and 13 of experiment, (3) decreased the expression of ovocalyxin-36 and ovocleidin-116 on the day 6 of experiment. The results obtained indicate that GH, by regulation of E2 concentration and ERs gene expression as well as expression of the egg shell-specific proteins produced in the shell gland, may determine the rate of regression, rejuvenation and functioning of this organ in chickens during molting. Supported by: NCN grant no. 2011/01/B/NZ4/03665 and DS-3243/KFiEZ. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of adding herb extract to water on rectal and radiated temperature of laying hens during the summer heat Ewa Sosnówka-Czajka, Iwona Skomorucha Department of Technology, Ecology and Economics of Animal Production, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: ewa.sosnowka@izoo.krakow.pl The aim of the study was to determine the effect of supplementing drinking water with an antioxidant herb extract of sage and common nettle during the summer heat on rectal and radiated temperature of laying hens kept in furnished cages. The experiment was conducted from May to the end of September on 435 Hy-Line laying hens, which were kept in furnished cages (750 cm2/bird) from 16 weeks of age and assigned to three experimental groups. Group 1 (control) received no supplements in drinking water. In groups 2 and 3, water drinkers were supplemented from 20 to 44 weeks of age with extracts of sage (Salvia officinalis L.) and common nettle (Urtica dioica L.) at 2 ml/l water, respectively. Birds were exposed to elevated air temperatures throughout rearing. Chickens received ad libitum standard diets based on layer concentrates. Birds had free access to feed and water throughout the study. During days of high air temperature (week 28 of rearing), 10 hens per group were measured for radi- ated temperature of feathered and unfeathered body parts and for rectal temperature. The same measurements were made 3 days after the temperature dropped to 20.1°C (week 29 of rearing). The results were statistically analysed by one-way analysis of variance and significant differences were estimated with Duncan’s multiple range test. Birds from experimental groups 2 and 3, which received herb extracts in water, exhibited a decrease in rectal temperature at 29 weeks of rearing (when temperature inside the house was around 20°C) compared to the measurements made at higher air temperatures (p ≤ 0.01). On the first day of high temperature, radiated temperature of both feathered and unfeathered parts was higher in control birds and in birds receiving the sage extract in water. In all the groups, radiated temperature was observed to decrease during the 29th week of rearing. 175 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of breed of laying hens on production results under organic conditions Ewa Sosnówka-Czajka, Iwona Skomorucha Department of Technology, Ecology and Economics of Animal Production, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: ewa.sosnowka@izoo.krakow.pl The act on organic agriculture recommends that preference should be given to native breeds characterized by relatively good health and adaptation to local climatic conditions. However, choosing the breed of hens for producing organic table eggs may largely determine the profitability of production, because bird productivity will be the main factor affecting the efficiency of organic farming. Therefore, the aim of the experiment was to evaluate the productivity of hens of different native breeds kept according to organic farming principles on a certified poultry farm. Subjects were 16-week-old pullets of conserved breeds originating from organically raised Greenleg Partridge and Rhode Island Red (RIR) laying hens. Layers were fed semi-intensively according to organic farming principles. Feed mixtures consisted of organic components. Throughout the experiment, birds had free access to feed and water. Pullets were kept on litter at a density of 6 birds/m2 of floor space and had access to green paddocks (4 m2/bird). The paddocks were equipped with shelters and drinkers. 176 Production results were monitored during the study. The results were statistically analysed by oneway analysis of variance and significant differences were estimated with Duncan’s range test. Until 28 weeks of rearing, Rhode Island Red hens were characterized by highly significantly higher egg production compared to Greenleg Partridge hens. Later in the egg production period, differences between the studied breeds were statistically significant. RIR hens were also characterized by lower feed intake per egg. Breed of laying hens had no effect on the health and mortality of organically raised layers. The results show that RIR hens kept in accordance with the rules of organic farming achieve better performance compared to Greenleg Partridge layers. As a consequence, Rhode Island Red laying hens should be the breed of choice for producing organic table eggs. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of housing system on some parameters of egg quality and egg-laying behaviour in laying hens Ewa Sosnówka-Czajka, Iwona Skomorucha, Eugeniusz Herbut Department of Technology, Ecology and Economics of Animal Production, National Research Institute of Animal Production, Balice near Krakow, Poland e-mail: ewa.sosnowka@izoo.krakow.pl There is evidence in the literature that laying hens in the free-range system relatively often lay eggs outside the nest, preferring secluded free-range sites. This leads to a higher incidence of dirty eggs. Other data suggests that the highest egg production and the smallest number of dirty eggs are obtained in the cage system. Therefore, the aim of the study was to evaluate the effect of housing laying hens in furnished cages and on litter with free-range access on some parameters of egg quality and on the choice of egg-laying site by hens. The experiment used Hy-Line commercial layers from 16 to 44 weeks of age. Hens were fed ad libitum standard diets based on layer concentrates. Throughout the experiment, birds had free access to water. Pullets were assigned to two experimental groups differing in the housing system: group 1 – furnished cages (750 cm2/bird), group 2 – litter (9 birds/m2) with access to grass paddocks (2.5 m2/ bird). Paddocks were equipped with drinkers and shelters providing protection from adverse weather. During the study, production results and the number of dirty, misshapen and cracked eggs were monitored. The number of eggs laid outside the nest was also recorded. The results were statistically analysed by one-way analysis of variance and significant differences were estimated with Duncan’s range test. Layers kept in the barn system with outdoor access were characterized by higher egg production throughout rearing compared to hens from the cage system. The number of eggs laid outside the nest was significantly higher for the furnished cage system compared to the free-range system. The number of cracked eggs was higher in the cage system compared to group 2. The housing system of Hy-Line commercial layers had no effect on the number of dirty and misshapen eggs. The results obtained show that rearing system influences the laying behaviour of hens and the number of cracked eggs, but does not necessarily determine the number of dirty eggs. 177 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The comparison of microbial transmission within intensive (commercial farm) and extensive housing systems on an annual basis Kinga Stuper-Szablewska1, Tomasz Szablewski2, Sebastian Nowaczewski3, Renata Cegielska-Radziejewska2 Department of Chemistry, Poznań University of Life Sciences, Poznan, Poland Department of Food Quality Management, Poznań University of Life Sciences, Poznan, Poland 3 Department of Animals Breeding and Product Quality Assessment, Poznań University of Life Sciences, Suchy Las, Poland 1 2 e-mail: sebnow@up.poznan.pl Respirable dust, which then as a result of agglomeration of the particles with the participation of water becomes part of the settled particulate matter is a large source of microbiological contamination. It includes next elements with irritation, inflammation and allergenic (eg. spores and fragments from mold). Removal of volatile impurities microclimate of the house is extremely important for both animal welfare and their health. This threat, however, has not yet been examined in relation to extensive housing systems in which hens are kept in a smaller cast with access to the runway and the control of the rearing conditions is greatly reduced. A comprehensive assessment of the microclimate of the house and the transmission of contaminants outside the building on an annual basis in terms of microbial contamination. They were compared two types of laying hens on litter: extensive and commercial housing systems. Dust inside the commercial 178 house contain more microorganisms than extensive system. Nevertheless, the size of microbiological contaminants contained in the dust collected inside the extensive henhouse not significantly different from the results obtained in intensive. The content of microflora in the dust settled on the outside of the house system intensive covered to a large extent (approx. 80%) with individual groups of microorganisms identified in the dust inside the house, which indicates a high transmission of microorganisms. Based on the results of microbiology dust taken from inside and outside the house intensive found that a large number of microorganisms is removed from the microclimate of the house thanks to the forced ventilation fans. In the case of the house extensive where the dust is not removed from its interior to a large extent, residual indoor livestock may be the cause of diseases of birds. Porównanie transmisji wyziewów mikrobiologicznych z kurników wielkotowarowych i przyzagrodowych w cyklu rocznym Kinga Stuper-Szablewska1, Tomasz Szablewski2, Sebastian Nowaczewski3, Renata Cegielska-Radziejewska2 Katedra Chemii, Uniwersytet Przyrodniczy w Poznaniu Katedra Zarządzania Jakością Żywności, Wydział Nauk o Żywności i Żywieniu, Uniwersytet Przyrodniczy w Poznaniu 3 Katedra Hodowli Zwierząt i Oceny Surowców, Wydział Medycyny Weterynaryjnej i Nauk o Zwierzętach, Uniwersytet Przyrodniczy w Poznaniu, Suchy Las 1 2 e-mail: sebnow@up.poznan.pl Pył respirabilny jest źródłem dużego zanieczyszczenia mikrobiologicznego. W jego skład wchodzą elementy mające działanie drażniące, zapalne oraz alergizujące (m.in. zarodniki i fragmenty grzybni pleśni). Usuwanie zanieczyszczeń lotnych mikroklimatu kurnika jest niezwykle ważne ze względu zarówno na dobrostan zwierząt, jak i ich zdrowotność. Zagrożenie to nie zostało jeszcze zbadane w odniesieniu do chowu przyzagrodowego, w którym utrzymywane są nioski w mniejszej obsadzie z dostępem do wybiegu, a kontrola warunków chowu jest znacznie ograniczona. Przeprowadzono kompleksową ocenę mikroklimatu kurnika oraz transmisji zanieczyszczeń na zewnątrz budynku w cyklu rocznym pod względem skażenia mikrobiologicznego. Porównane zostały dwa rodzaje chowu kur niosek na ściółce: przyzagrodowy i wielkotowarowy. Pył wewnątrz kurnika wielkotowarowego zawierał większą liczbę mikroorganizmów niż w systemie przyzagrodowym. Niemniej wielkości zanieczyszczeń mikrobiologicznych zawartych w pyle pobranym wewnątrz kurnika w chowie przyzagrodowym nie różniły się znacznie od wyników uzyskanych w chowie wielkotowarowym. Zawartość mikroflory w pyle osiadłym na zewnątrz kurnika w systemie wielkotowarowym pokrywała się w dużej mierze (ok. 80%) z wartościami poszczególnych grup mikroorganizmów zidentyfikowanych w pyle wewnątrz kurnika, co wskazuje na dużą transmisje mikroorganizmów. Na podstawie uzyskanych wyników mikrobiologii pyłu pobranego z wnętrza oraz z zewnątrz kurnika wielkotowarowego stwierdzono, że duża ilość mikroorganizmów jest usuwana z mikroklimatu kurnika dzięki wymuszonej wentylatorami wentylacji. W przypadku kurnika przyzagrodowego pył nie jest usuwany z jego wnętrza w istotnym stopniu, zalega wewnątrz pomieszczenia inwentarskiego i może być przyczyną chorób ptaków. 179 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The evaluation of the effect of seasons on the microbiological contamination of selected elements of the environment in terms of health safety of laying hens kept in extensive housing system Kinga Stuper-Szablewska1, Tomasz Szablewski2, Sebastian Nowaczewski3, Anna Ostrowska-Kołodziejczak1, Anna Matysiak1, Renata Cegielska-Radziejewska2 Department of Chemistry, Poznań University of Life Sciences, Poznan, Poland Department of Food Quality Management, Poznań University of Life Sciences, Poznan, Poland 3 Department of Animals Breeding and Origin Materials, Poznań University of Life Sciences, Suchy Las, Poland 1 2 e-mail: sebnow@up.poznan.pl The aim of an experiment was the evaluation of the effect of seasons on the microbiological contamination of selected elements of the environment in terms of health safety of laying hens kept in extensive housing system. The study was conducted to analyse of microbial contamination of all environmental elements (feed, litter, dust and microclimate of the henhouse). Research was carried out on an annual basis. The samples were taken from 5 henhouses with a small stocking density of birds (less than 1 per m2) and with avail- 180 able gritty run. The total number of aerobic bacteria (OLB), Pseudomonas and Enterobacteriaceae genus, as well as microscopic fungi (qualitatively and quantitatively) were determined. The season had a significant impact on microbiological contamination of the environment and the microclimate of the henhouse. The results showed that spring and autumn are the seasons conducive to development of mold. On the other hand, autumn and winter are characterized by a significantly higher bacterial contamination. Ocena wpływu pory roku na zanieczyszczenie mikrobiologiczne wybranych elementów środowiska kurnika w aspekcie bezpieczeństwa zdrowotnego kur niosek utrzymywanych w systemie przyzagrodowym Kinga Stuper-Szablewska1, Tomasz Szablewski2, Sebastian Nowaczewski3, Anna Ostrowska-Kołodziejczak1, Anna Matysiak1, Renata Cegielska-Radziejewska2 Katedra Chemii, Uniwersytet Przyrodniczy w Poznaniu Katedra Zarządzania Jakością Żywności, Wydział Nauk o Żywności i Żywieniu, Uniwersytet Przyrodniczy w Poznaniu 3 Katedra Hodowli Zwierząt i Oceny Surowców, Wydział Medycyny Weterynaryjnej i Nauk o Zwierzętach, Uniwersytet Przyrodniczy w Poznaniu, Suchy Las 1 2 e-mail: sebnow@up.poznan.pl W Polsce od wielu lat utrzymuje się liczna grupa konsumentów zainteresowana nabywaniem jaj z chowu przyzagrodowego, ocenianych jako żywność wysokiej jakości i jednocześnie bezpieczna dla zdrowia, ale tańsza niż żywność ekologiczna, potwierdzona certyfikatem. Zapewnienie dobrostanu kur niosek utrzymywanych w systemie przyzagrodowym jest tematem często pomijanym w odniesieniu do innych, wielkotowarowych systemów chowu. Jednym z istotnych czynników związanym z dobrostanem niosek jest ich zdrowotność, na którą niewątpliwie wpływ ma poziom zanieczyszczenia mikrobiologicznego środowiska kurnika. Źródłem patogennej mikroflory oraz mykobiota mogą być: pasza, ściółka, gleba, pył, woda oraz elementy budynków inwentarskich. Prowadzone badania stanowiły kompleksową analizę zanieczyszczenia mikrobiologicznego wszystkich elementów tworzących środowisko kurnika (pasza, ściółka, pyły oraz elementy mikroklimatu kurnika), w którym utrzymywano nioski w systemie przyzagrodowym. Badania prowadzone były w cyklu rocznym. Próby pobierano z 5 kurników z niewielką obsadą ptaków (mniej niż 1 szt./m2) mających do dyspozycji piaszczysty wybieg. Nioski, mieszańce ROSA, w mo- mencie rozpoczęcia pobierania prób miały 20 tygodni życia. W kurniku materiał ściołowy stanowiła słoma. Próby pyłu osiadłego (w kurniku i na zewnątrz), paszy oraz ściółki zostały poddane badaniom mikrobiologicznym. Oznaczano ogólną liczbę bakterii tlenowych (OLB), bakterie z rodzaju Pseudomonas oraz Enterobacteriaceae, a także ilościowo i jakościowo grzyby mikroskopowe. Na podstawie przeprowadzonych badań stwierdzono, że pory roku miały istotny wpływ na zanieczyszczenie mikrobiologiczne elementów środowiska oraz mikroklimatu kurnika. Uzyskane wyniki wykazały, iż wiosna i jesień są porami roku sprzyjającymi rozwojowi grzybów pleśniowych, natomiast jesień i zima cechują się istotnie wyższym wzrostem zanieczyszczenia bakteriami szczególnie z grupy Enterobacteriaceae. Wśród badanych elementów środowiska kurnika ściółka i pył były najbardziej zanieczyszczone. Analiza dyskryminacyjna całościowego zanieczyszczenia 5 badanych kurników wykazała istotne różnice w zanieczyszczeniu mikrobiologicznym badanych obiektów i wskazała, że to pył (zarówno respirabilny, jak i osiadły) ma największy wpływ na stan mikrobiologiczny kurnika. 181 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The effect of methionine in ovo administration on chick (Gallus gallus domesticus) embryo development Barbara Tombarkiewicz, Karolina Trzeciak, Marcin Lis, Bartosz Bojarski, Krzysztof Pawlak, Weronika Kanik, Zbigniew Podgórni, Jerzy Niedziółka Department of Veterinary Science, Animal Reproduction and Welfare, Institute of Veterinary Sciences, University of Agriculture in Krakow, Poland e-mail: rztombar@cyf-kr.edu.pl The aim of this study was to determine the impact of various doses of methionine administered at early stage of embryogenesis on the course of and postnatal development, as well as selected hematological and biochemical blood parameters of the domestic chicken embryo (Gallus gallus domesticus). The experimental material constituted 125 eggs from the broiler breeder flock of Ross 308 line divided on five groups: control group (O), shame group (K) – injected in ovo by physiological saline and three experimental groups, which were injected by 5 mg, 25 mg and 50 mg of methionine. Biochemical (the concentration of uric acid and homocysteine in the blood plasma and the glucose in whole blood) and hematological (blood hemoglobin content, hematocrit value, RBC, WBC with leukogramem, MCV, MCH, MCHC) analyzes of blood were performed as well as the histological analyzes of selected organs. 182 It was proved that excess of methionine may interupt the embryogenesis process and cause malformations and also affect the histological image of pancreas, kidney and liver. The administration of methionine at 4th day of embryogenesis increases mortality immediately after manipulation. The chick kidney microstructure in the first day after hatching (D1) contain both regular renal corpuscules, as well as impaired-structure ones. In the pancreases of chicks injected by the highest dose of methionine a lot of large vacuoles were observed in pancreases cells, which may indicate on fatty degeneration of this organ. Any hematological nor biochemical changes of blood parameters were noted. This research was financially supported by the statutory activity: DS 3263. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Effect of probiotic application as a spray to egg shell on hatchability of broiler ducks Ján Weis, Cyril Hrnčár, Matej Rindoš, Slavomír Mindek Department of Poultry Science and Small Animal Husbandry, Slovak University of Agriculture in Nitra, Slovakia e-mail: jan.weis@uniag.sk The aim of this study was to evaluate the effect of application of Lactobacillus fermentum as a spray on hatchability of broiler ducks. Totally 160 hatching eggs were divided to two groups. In experimental group (n = 80) we applied probiotic strain Lactobacillus fermentum CCM 7158 with concentration of 1x109 colony forming units (CFU) as a spray to egg shell on 24th day of incubation. The control group (n = 80) was without any application. A total egg hatchability, hatchability of fertile eggs set, dead in shells and saleable ducks were measured. Application of probiotic strain Lactobacillus fermentum caused statistically significant increase(P < 0.01) total hatchability, hatchability of fertilised eggs, saleable ducks and statistically significant (P < 0.01) decreased dead in shells. 183 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Changes in thyroid hormone concentrations in chick blood during hatching after in ovo α tocopherol administration Gabriela Wójcik1, Magdalena Trela1, Andrzej Sechman2, Marcin W. Lis1 1 Department of Veterinary, Animal Reproduction and Welfare, University of Agriculture in Krakow, Poland 2 Department of Animal Physiology and Endocrinology, University of Agriculture in Krakow, Poland e-mail: rzlis@cyf-kr.edu.pl Plasma thyroid hormone concentration in the chick embryo sharply increases toward the end of the incubation. It results from mobilization of the organism to hatching, associated with the yolk sac retraction, the onset of pulmonary respiration and initiation of endothermic responses. The hatching is also very stressful and tiring for the chick, therefore there are attempts to improve of chick health by an in ovo supplementation of the chick embryo with antioxidative vitamins such as ascorbic acid (vitamin C) or tocopherols (vitamin E). Since the E hypervitaminosis can inhibit the thyroid gland activity in mammals and birds, it is interesting to investigate the effect of in ovo administration of α-tocopherol (α-Toc) on thyroid hormone concentrations in the blood plasma of the chick embryo during hatching. At the 17th day of incubation the embryonic chicken eggs (broiler line Ross 308, n = 40 eggs/ group) were injected with 0 (control), 0.5 or 5.0 mg α-Toc/egg dissolved in 0.1 ml arachid oil. Blood samples were collected on embryonic on the day 18 (E18), during internal (IP) and external pipping (EP), and from hatched chicks (H) and at 6–12 hours after the hatching (D1). The concentrations of thyroxine (T4) and triiodothyronine (T3) in plasma samples were 184 determined by RIA method. Results were statistically evaluated by a two-way ANOVA followed by Tukey test at p < 0.05. In the control group, the highest levels of T4 and T3 were found during IP (7.7 and 10.4 nmol/L, respectively). Following the IP, a rapid decrease in these hormones was observed. In comparison to the control group, the administration of 0.5 and 5.0 mg α-Toc reduced T4 and T3 plasma concentrations at the IP by 58% and 70% (P ≤ 0.05), and 54% and 50% (P ≤ 0.05), respectively. Simultaneously, T4 and T3 levels in blood of the H were more than 25% higher in comparison to control group (P ≤ 0.05). Moreover, the calculated T3/T4 ratio was similar in the control and in the 0.5 mg α-Toc treated chicks during the whole hatching period (1.2–1.4); in the experimental group treated with the higher dose of α-Toc an increase in T3/T4 rate was observed (2.1–3.1; P ≤ 0.05). In summary, the increased availability of α-tocopherol at the end of the embryogenesis may affect the synthesis of thyroid hormones in hatching chicks. Studies supported by DS ZWRiDZ/2016. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Comparison of parasite infection in laying hens kept in different housing systems Anna Wyrobisz1, Ewa Sosnówka-Czajka2, Jerzy Kowal1, Paweł Nosal1 Department of Environmental Zoology, Institute of Animal Sciences, University of Agriculture in Krakow, Poland 2 Department of Technology, Ecology and Economics of Animal Production, National Research Institute of Animal Production, Balice near Krakow, Poland 1 e-mail: a.wyrobisz@gmail.com Welfare of laying hens depends on many overlapping factors. In particular, poultry farming should provide appropriate housing system that may have influence on health condition. Many diseases pose a serious threat to the poultry welfare, however, ailments caused by parasites should be regarded as very essential. Furthermore, the range of their occurrence can be related to the housing system. The aim of the presented study was to compare the parasite infection level in commercial line of laying hens from different housing systems. The research focused on Hy-Line hens housed in barn, barn with paddock or enriched cage system. Parasitic fauna composition, as well as the level of infection was determined by ante-mortem (coproscopical) and post-mortem (dissection) analyses. Faeces samples were examined twice – before (November) and after (March) winter time. The coproscopical analysis revealed the presence of coccidian oocysts (Eimeria sp.) and nematode eggs (Ascaridia sp., Heterakis sp.), whereas in dissected material only nematode (Ascaridia sp.) specimens were found. The hens kept in a barn were slightly more infected with Eimeria sp. than these with access to a paddock. In contrast, the majority of analyzed animals from barn with paddock system were infected with Ascaridia sp. Both the prevalence and intensity of the infection reached high values. Despite stated differences in nematode infection between each housing system in coproscopical analysis, postmortem results were not consistent with them. Only hens kept in enriched cages were parasites-free. The obtained results indicate that the level of infection with parasites from different groups (coccidia or nematodes) is highly related to the used housing system. Although, the enriched cage housing system is considered to be the least animal-friendly, parasitological approach has contradicted it. Hens kept in barn, as well as barn with paddock system should be routinely controlled, as a part of animal welfare. 185 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 The method of identifying species of poultry on the basis of sequence differences in the s-rRNA Małgorzata Natonek-Wiśniewska, Piotr Krzyścin Department of Genomics and Molecular Biology of Animals, National Research Institute of Animal Production, Balice near Krakow e-mail malgorzata.natonek@izoo.krakow.pl The EU regulations require manufacturers of food and animal feed accurate information about the qualitative composition of the product. Therefore, to develop and implement methods to verify the declared composition of the real has become an important challenge in recent years. The aim of this study was to develop a method to quickly determine the presence of poultry: chicken (Gallus gallus), ducks (Anas platyrhynchos), geese (Anser anser), turkeys (Meleagris gallopavo) and guinea fowl (Numida meleagris) based on the analysis of the region s-rRNA mitochondrial DNA. The research material was a sample of meat five species of poultry from which the DNA was isolated using a set of Axfood (A & A Biotechnology). 186 Using the Primer3 program primers were designed: 5’CCCACCTAGAGGAGCCTGTT and 5’GGCGGTATGTAGGCTGTGTT giving a PCR product with a size of 81 bp. Designated starters are characterized by high species specificity ‒ they are homologous to the species for which they were designed, while other species of farm animals show no similarities. The proposed method allows the identification of the poultry in one PCR reaction which significantly reduces the time and cost of a separate determination of each species. Introduction of the practice of laboratory developed methods contribute to the implementation of the EU requirements regarding the qualitative analysis of food products and animal feed. International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Fatty acid profile of intramuscular fat in supraspinatus, longissimus lumborum and semimembranosus muscles of farmed fallow deer Urszula Kaczor1, Marek Stanisz2, Marta Bykowska2, J. Składanowska2, Dorota Wojtysiak3, Andrzej Kaczor4 Department of Animal Biotechnology, University of Agriculture in Krakow, Poland Department of Animal Breeding and Product Quality Assessment, Poznań University of Life Sciences, Poland 3 Department of Animal Anatomy, University of Agriculture in Krakow, Poland 4 Department of Technology, Ecology and Economics of Animal Production, The National Research Institute of Animal Production, Balice near Krakow, Poland 1 2 e-mail: rzkaczor@cyf-kr.edu.pl The world population of farmed Cervidae continues to grow and now exceeds 5 million. Meat production is the most popular purpose of Cervidae farming in Poland and around the world. The high nutritive value of their meat, which is high in protein and minerals and low in fat and cholesterol, is greatly appreciated. The aim of the study was to analyse the intramuscular fat content and fatty acid profile of supraspinatus (SS), longissimus lumborum (LL) and semimembranosus (MS) muscles in ten 18-month-old male fallow deer (Dama dama) raised in an organic farm located in north-western Poland. The analysed muscles had a low content of intramuscular fat: 0.73% (LL) and 0.77 (MS) vs 1.21% (SS) (P < 0.01). In the analysed muscles, stearic acid C18:0 and palmitic acid C16:0 dominated among saturated fatty acids (SFA), while linolenic acid C18:2 among polyunsaturated fatty acids (PUFA). The fatty acid profile of LL and MS dif- fered significantly from that found for SS. SFA content of the above muscles was 33.3% and 33.8% vs 43.6%, respectively (P < 0.01) (% of total acids determined). PUFA content was high at 46.6% and 48.1% vs 34.1%, respectively (P < 0.01). n-3 PUFA values for LL and MS vs. SS were 12.8% and 13.9% vs 11.% (P < 0.01), and the atherogenic index (IA) was 0.23 and 0.24 vs 0.39 (P < 0.01). The content of hypocholesterolemic acids (DFA) and hypercholesterolemic acids (OFA) in SS differed significantly from that in LL and MS. At the same time, the lowest DFA/OFA values were found for SS muscle. The results obtained show the high nutritive value of fallow deer meat, but the location of adipose tissue causes significant differences in the fatty acid profile. Meat sampled from the shoulder (supraspinatus muscle) has a higher content of intramuscular fat but shows a less favourable profile of the analysed fatty acids. 187 International Conference on Biotechnology and Welfare in Animal Science with a session on “7th Poultry Days” • Krakow • 23–24 June 2016 Histochemical muscle fibre characteristics of fallow deer meat Dorota Wojtysiak1, Marek Stanisz2, Agnieszka Ludwiczak2, Urszula Kaczor3 Department of Animal Anatomy, University of Agriculture in Krakow, Poland Department of Animal Breeding and Product Quality Assessment, Poznań University of Life Sciences, Poland 3 Department of Animal Biotechnology, University of Agriculture in Krakow, Poland 1 2 e-mail: wojtysiakd@wp.pl In recent years fallow deer farming has become extremely popular. Fallow deer meat is characterized by a specific taste of meat. The unique properties of fallow deer meat are caused by a low content of intramuscular connective tissue and a different histochemical muscle fibre composition compared to farm animals. It is well known that muscle fibre type composition of different skeletal muscles may be associated with post-mortem changes in the conversion of muscle to meat and subsequently meat quality. Therefore, the aim of this study was to compare fibre type composition in semimembranosus, supraspinatus, and longisimus lumborum muscles of fallow deer. A total of 10 carcasses from male of fallow deer (Dama dama) shot at 18 months during winter in the farm of the Złotowski district were evaluated. Shortly after being shot muscle samples were taken from the mid-part of semimembranosus, supraspinatus, and longisimus lumborum muscles and frozen in liquid nitrogen. To determine the muscle fibre types, we used a modified combined method of NADH-tetrazolium 188 reductase activity and immunohistochemical determination of the slow myosin heavy chain izoforms. The results obtain indicated that semimembranosus, supraspinatus, and longisimus lumborum muscles have a heterogenic structure and consists of mixture of fibre types having different contractile and metabolic properties namely, type I (slow-twitch oxidative), IIA (fast-twitch oxidative) and IIB (fast-twitch glycolytic) fibres. Moreover, analysis of the muscle fibre composition showed that muscle type has a significant effect on both muscle fibre percentage and size. Thus, the supraspinatus muscles of fallow deer are the most oxidative, as evidenced by the highest percentage of type I fibres compared to the longissimus lumborum and semimembranosus muscles. Regarding muscle fibre size, the longissimus lumborum muscle has significantly the smallest muscle fibre types I and IIA than other muscles. Funded by DS-3253. Index of authors A Adaszyńska-Skwirzyńska Michalina 123, 124 Ancsin Zsolt 133, 157 Anderle Vojtěch 151 Andraszek Katarzyna 96 Andres Krzysztof 136, 137, 154 Antonova Ekaterina P. 79, 112 Antończyk Anna 97 Antos Piotr A. 21 Anusz Krzysztof 73 Arpášová Henrieta 121, 140 Augustyn Romana 31, 32, 45, 46, 54, 55, 56, 57, 81, 82, 108 B Babicz Marek 110, 111 Baishnikova Irina V. 79, 91, 112 Bakst Murray 154 Balogh Krisztián 125, 133, 157, 161 Baranowska Agnieszka 83 Barszcz Karolina 135, 144 Bauer Edyta 75, 76 Bednarczyk Marek 122 Bilčík Boris 155, 162 Bilińska Barbara 88, 90, 159 Blicharski Tadeusz 110, 111 Błasiak Michał 98 Bocheńska Aneta 84 Bodnar Julia 113 Bojarski Bartosz 160, 182 Borkowski Tomasz 168 Borodulina Victoria I. 28 Bócsai Andrea 125, 133, 157 Bucław Mateusz 126, 127 Bujko Jozef 140 Buríková Monika 155 Bykowska Marta 187 C Calik Jolanta 128, 129, 130, 155 Cebulska Aleksandra 34 Cegielska-Radziejewska Renata 178, 179, 180, 181 Chałupka Daniela 145, 167, 171 Chernobrovkina Nadezhda P. 112 Chоkаn Taras 85 Chyb Jarosław 86, 105 Ciereszko Andrzej 99, 100, 159 Ciesielczyk Katarzyna 83 Cora Gabriela 86 Čavarga Ivan 155 Čoudková Veronika 30 D Dąbrowski Sebastian 106 Demey Vanessa 58, 60 Dietrich Mariola A. 99, 100 Długosz Bogusława 31, 32, 45, 46, 54, 55, 56, 57, 81, 82, 108 Dobosz Iwona 87 Dobrzańska Kinga 45, 46 Domagała Justyna 83 Drąg-Kozak Ewa 33, 49, 50, 105 Duda Małgorzata 101, 114 Dulińska-Litewka Joanna 88 Dunisławska Aleksandra 122 Dybała Jan 34 Dykiel Magdalena 131 Dżugan Małgorzata 132 E Erdélyi Márta 133, 157 F Faundez Ricardo 106 Fernye Csaba 133 Fic Kinga 35, 102, 107 Filipczyk Anna 107 Flaga Jadwiga 58, 60 Foltyn Marian 134 Frankowska Katarzyna 107 G Gajewska Marta 106 Gąsior Robert 129 Gertler Arieh 15 Gorczyca Gabriela 114 189 Gosiewski Grzegorz 33 Goździewska-Harłajczuk Karolina 135, 144 Górowska-Wójtowicz Ewelina 88, 90, 159 Górska Magdalena 74, 136, 137 Graczyk Magdalena 136, 137 Grzesiak Małgorzata 143 Grzesiakowska Anna 96 Guja Iwona 36, 44, 103 H Halek Anna 54, 55 Halik Gabriela 89 Hanus Anton 138 Hanusová Emília 138 Hartwich Patryk 102 Hejmej Anna 88, 90, 159 Herbut Eugeniusz 177 Hereta Marta 114 Hołownia G. 99, 100 Horecka Eliska 139 Horecky Cenek 139 Horváth Mária 146 Hrabia Anna 21, 174 Hrnčár Cyril 121, 138, 140, 183 I Ilyina Tatiana N. 79, 91 Ilyukha Viktor A. 44, 79, 91, 112 Izmailovich Inessa B. 141 J Jackowska-Tracz Agnieszka Jagusiak Wojciech 65 Jaklińska Barbara 31, 32 Jozkowicz Alicja 80 Jóźwiak Michał 142 Jura Jacek 116 73 K Kaczor Andrzej 37, 38, 93, 187 Kaczor Urszula 37, 38, 93, 94, 95, 187, 188 Kamińska Alicja 88, 90, 159 Kanik Weronika 160, 182 Kapelański Wojciech 34 Kapkowska Ewa 136, 137 Karasiński Janusz 115, 116 Katarzyńska Dorota 21, 143 Kawęcka Aldona 39, 40, 41 Kądzior Justyna 56, 57 Kermen-Germaniuk Katarzyna 106 Khizhkin Evgeny A. 79 Kirsz Katarzyna 84, 92, 109 Kizhina Aleksandra G. 79 Klećkowska-Nawrot Joanna 135, 144 Knapczyk-Stwora Katarzyna 87 Knoll Ales 139 Kokocińska Agata 42, 43 Kołacz Roman 16 Kołodziejski Paweł A. 145, 167, 171 190 Korytkowski Łukasz 58, 60 Košťál Ľubor 146, 162 Kotula-Balak Małgorzata 88, 90, 159 Kovács Balázs 161 Kovarikova Lenka 139 Kövesi Benjámin 161 Kowal Jerzy 185 Kowalczyk Artur 144, 154, 169, 170 Kowalski Zygmunt M. 58, 60 Koziorzębska Agata 89 Krawczyk Józefa 147, 148, 149, 150 Król Teodora 132 Krzystanek Agnieszka 35 Krzyścin Piotr 48, 186 Kucharski Mirosław 93, 94, 95 Kuchta-Gładysz Marta 96, 103 Kupčíková Lucie 151 Kuzmina Natalia 113 L Lasoń Martyna 153, 154 Latacz Anna 117 Lichovníková Martina 134, 151, 152 Lipień Gabriela 103 Lis Marcin W. 79, 132, 141, 160, 182, 184 Lisowski Piotr 86 Luber Magdalena 93 Ludwiczak Agnieszka 188 Ł Łapiński Stanisław 36, 44, 79, 91, 103, 112 Łozicki Andrzej 89 Łukaszewicz Ewa 144, 153, 154, 169, 170 Łuszczek-Trojnar Ewa 33, 49, 50 Łuszczyński Jarosław 31, 32, 45, 46, 54, 55, 56, 57, 81, 82, 108 M Maćkowiak Paweł 145, 167, 171 Máčajová Mariana 155 Maiorano Giuseppe 122 Makarski Mateusz 89 Malkiewicz Marta 58, 60 Maršálek Miroslav 30 Matusiewicz Magdalena 89 Matysiak Anna 180, 181 Mézes Miklós 125, 133, 157, 161 Mika Magdalena 97 Mika Maria 174 Miksza-Cybulska Anna 39 Mikulich Elena L. 47 Milewska Aleksandra 34 Mindek Slavomír 183 Minta Zenon 142 Misztal Tomasz 98 Molik Edyta 98 Mostek Agnieszka 99, 100 Motal Maria 101 Mrozicka Katarzyna 44 Mucha Aurelia 110, 111 Muchacka Renata 156 N Nakade Mangesh 157 Nalik Klaudia 102, 107 Natonek-Wiśniewska Małgorzata 48, 186 Niedbała Piotr 36, 44, 79, 96, 103, 112 Niedziółka Jerzy 132, 160, 182 Niemczuk Krzysztof 166 Niwińska Anna 106 Nogowski Leszek 145, 167, 171 Nosal Paweł 185 Nowacki Patryk 49, 50 Nowaczewski Sebastian 178, 179, 180, 181 Nowak Jakub 132 Nowak Joanna 158, 165 Nowakowski Michał 102 O Obrzut Joanna 130, 148 Ochrem Andrzej 75, 76 Ocłoń Ewa 117 Olszewska-Tomczyk Monika 142 Oravcová Marta 138 Ostapiv Dmytro 113 Ostrowska-Kołodziejczak Anna 180, 181 Ostrowski Jerzy 106 Otwinowska-Mindur Agnieszka 51, 96 P Paraponiak Paweł 27, 52, 53 Pardyak Laura 88, 90, 159 Pasternak Marta 104 Pastuszak Karolina 94 Pavlík Ales 139 Pawlak Krzysztof 160, 182 Pelyhe Csilla 125, 161 Petrajtis-Golobow Monika 106 Pichová Katarína 146, 162 Pieszka Magdalena 31, 32, 45, 46, 54, 55, 56, 57, 81, 82, 108 Pietsch-Fulbiszewska Agnieszka 84 Piórkowska Katarzyna 94, 95, 108, 110, 158 Pisarczyk Weronika 31, 32, 46, 54, 55, 56, 57, 81, 82, 108 Pistová Veronika 121, 140 Płowiec Arkadiusz 122 Podbielska Angelika 163, 164, 168 Podgórni Zbigniew 182 Podstawski Zenon 31, 32, 45, 46, 54, 55, 56, 57, 81, 82, 108 Połtowicz Katarzyna 129, 158, 165 Popek Włodzimierz 33, 49, 50, 105 Porubský Denis 140 Posyniak Andrzej 166 Próchniak Tomasz 31, 32, 81, 82 Pruszyńska-Oszmałek Ewa 145, 167, 171 Przywarová Alena 151 Ptak Ewa 51, 65 Puchała Michał 39, 66, 67, 130, 149, 150 Pustuła A. 81, 82 Pyś Jan B. 58, 60 R Rada Vojtěch 134 Radko Anna 85, 163, 164, 168 Radkowska Iwona 61, 62, 63, 64 Radkowski Adam 62, 63, 64 Rindoš Matej 183 Romanowicz Katarzyna 98 Romek Marek 116 Ropka-Molik Katarzyna 108, 110, 111 Rosenberger Joanna 154, 169, 170 Rubiś Dominika 85 Rzońca Zenon 169, 170 S Sadomov N.A. 28 Sandowska-Markiewicz Zuzanna 106 Sassek Maciej 145, 167, 171 Satoła Alicja 65 Sechman Andrzej 21, 143, 174, 184 Sergina Svetlana N. 79, 91, 112 Sharan Mykola 113 Sikora Jacek 40 Siwek Maria 122 Składanowska J. 187 Skomorucha Iwona 156, 173, 175, 176, 177 Skowron Beata 83 Skrzyński Grzegorz 75, 76 Sławińska Anna 122 Słomski Ryszard 116 Słowińska Mariola 159 Smorąg Zdzisław 116 Šoch Miloslav 68 Socha Joanna 174 Socha Magdalena 33, 49, 50, 86, 105 Sokołowicz Zofia 131 Sokołowska-Mikołajczyk Mirosława 86, 105 Sokołowski Grzegorz 106 Sosin-Bzducha Ewa 41, 66, 67 Sosnówka-Czajka Ewa 156, 172, 173, 175, 176, 177, 185 Sowinski Piotr SJ 25, 26 Sowińska Natalia 35, 84, 107 Sowula-Skrzyńska Elżbieta 75, 76 Stadnicka Katarzyna 122 Stanisz Marek 187, 188 Starý J. 30 Starzyńska-Janiszewska Anna 97 Stefaniuk-Szmukier Monika 31, 32, 45, 46, 54, 55, 56, 57, 81, 82, 108 Stręk Paweł 102 Stuper-Szablewska Kinga 178, 179, 180, 181 Szablewski Tomasz 178, 179, 180, 181 Szabó-Fodor Judit 125 Szczerbińska Danuta 123, 124 191 Szczęsna Małgorzata 92, 109 Szczęśniak Katarzyna 90 Szeleszczuk Olga 96, 103 Szmotała Tomasz 108 Szumiec Agnieszka 85,163, 164, 168 Szwaczkowski Tomasz 136, 137, 145, 167, 171 Szyndler-Nędza Magdalena 110, 111 Ś Śmietanka Krzysztof 142 Świderska Karina 31, 32 Świętoń Edyta 142 T Tarasjuk Serhiy 85 Tejml Petr 68 Thor Piotr J. 83 Tokodi Nikolett 125 Tombarkiewicz Barbara 160, 182 Tomczyk-Wrona Iwona 69, 71 Tracz Michał 73 Trapezov Oleg V. 79 Trela Magdalena 184 Trzeciak Karolina 182 Tyl-Bielicka Anita 106 Tyutyunnik Nikolay N. 112 U Unzhakov Aleksey R. 112 Uzenbaeva Lyudmila B. 79, 112 V Vlizlo Vasyl 113 Volna Sarka 152 Výboh Pavel 155 W Wartalski Kamil 101, 114 Weis Ján 140, 183 Wesołowska Monika 132 Westfalewicz Błażej 99, 100 Wiater Jerzy 115, 116 Wikiera Agnieszka 97 Wiśniewska Joanna 34 Wojciechowska Joanna 74 Wojtysiak Dorota 74, 187, 188 Woszczenko Aleksandra 35 Woszczyński Marek 106 Wójcik Gabriela 184 Wyrobisz Anna 185 Wyrostek Krzysztof 142 X Xu Yongping 79 Y Yaremchuk Іryna 113 Z Zándoki Erika 125, 157 Zemsta Katarzyna 95 Zięba Dorota A. 92, 98, 109 Zubel-Łojek Joanna 117 Ż Żychlińska-Buczek Justyna 75, 76