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Studies on Turkey Parvoviruses Dissertation

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Studies on Turkey Parvoviruses Dissertation STUDIES ON TURKEY PARVOVIRUSES DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Maria Vittoria Murgia Graduate Program in Veterinary Preventive Medicine The Ohio State University 2012 Dissertation Committee: Dr. Y. M. Saif (Advisor) Dr. D. J. Jackwood Dr. C-W Lee Dr. J. LeJeune Copyrighted by Maria Vittoria Murgia 2012 Abstract Turkey parvovirus belongs to the family Parvoviridae, subfamily Parvovirinae, genus parvovirus. It was identified in turkeys with enteritis for the first time in 1983. Since then there were no further reports on turkey parvovirus in the United States (US) until 2008 when Zsak et al. determined the partial sequence of the non structural (NS) gene of chicken and turkey parvoviruses and then in 2009 developed a PCR which was used to test fecal samples collected from various US states. A high prevalence of parvoviruses was detected by PCR in that study; however there was no information on the health status of the flocks from which the samples were collected. Moreover, there is no information regarding the pathogenesis of turkey parvoviruses and their involvement in the enteric diseases of poultry. The objectives of our studies were: 1. To determine the prevalence of parvovirus in various US states in different years 2. To determine the presence of other viruses in turkey poults in conjunction with parvoviruses 3. To develop a sensitive diagnostic assay for the detection of turkey parvoviruses in fecal samples 4. To determine the pathogenicity of parvovirus in SPF turkey poults. Intestinal content, feces or litter samples collected between 2000 and 2010 in four different states (Virginia, North Carolina, Pennsylvania, and Ohio) were used to address ii the first two objectives. Those samples were tested for parvovirus and other enteric viruses using PCR and RT-PCR with previously published primers and also with transmission electron microscopy. The overall prevalence of parvovirus (71.5%) was comparable with the previous studies. In our study, we had samples collected from birds of a wider age range and we compared the prevalence of parvoviruses in two age groups, 1-7 weeks and 8-19 weeks of age. We found that although parvovirus was widespread in both age groups, a significantly higher prevalence was detected in the older birds. Moreover, in the majority of the cases, parvovirus was detected in conjunction with other enteric viruses, such as astrovirus, reovirus and rotavirus. To support future surveillance and research, we develop Real-Time PCR test targeting a conserved region of the NS gene. This test showed the same analytical sensitivity and specificity compared to conventional PCR test and good intra and inter-assay repeatability. Moreover, it was shown to detect parvovirus’ DNA in field fecal samples. The real-time PCR is faster and it is possible to quantify the DNA which are advantages compared to the conventional PCR. Since we were not able to purify parvovirus, we determine the pathogenesis of parvovirus in combination with astrovirus and compare the outcome to that of astrovirus alone. In two different in vivo trials using 2-4 week-old SPF turkeys, we observed early replication of astrovirus in intestine and the parvovirus was detected in later time points when astrovirus shedding decreased. Our study shows potential viral interference among different enteric viruses and also shed light into the persistence or higher prevalence of parvovirus in turkeys of older ages in the field compared to astrovirus. iii Dedicated to my parents, my brother and my sister iv Acknowledgments I am very obliged to my advisor Dr. Yehia M. Saif for his guidance and support throughout my studies. He showed me how to look at everything with a positive attitude. He had always words of encouragement especially when the experiments did not work as expected. While working in his lab under his supervision I have grown as a scientist as well as a person and I will treasure this great experience for all of my life. I am grateful to my committee members Dr. Daral J. Jackwood, Dr. Chang-Won Lee, and Dr. Jeff LeJeune for their suggestions and comments. I am very thankful to Dr. Qiuhong Wang for her precious suggestions and comments. She always kept her door open when I needed her opinion. I would like to thank my labmates Dr. Abdul Rauf and former labmates Dr. Hadi Yassine and Dr. Yuxin Tang for their technical support and friendship. I am very thankful to Dr. Alex Rodriguez-Palaci and Dr. Kwonil Jung for their technical support and suggestions. I would like to thank Dr. Juliette Hanson, Kingsly Belin, Andrew Wright, Greg Myers, and Todd Root for their help with the animal care. I am very thankful to Ms. Robin Weimer and Ms. Hannah Gehman for their kindness and help during all these years. I am thankful to my fiancé Christian Cruz and his family for sharing with me this experience v Last but not least, I am obliged to my parents, Ms. Anna Franca Diana and Mr. Severino Murgia, by brother, Francesco, and my sister, Valeria, for supporting always my choices and for being close to me even if there is an ocean that physically separates us. I am appreciative to my aunt Mrs. Silvana Murgia, my grandmas Ms. Maria Mameli and Ms. Bonaria Argiolas for their encouragements throughout my life. I also would like to remember my grandpa Mr. Salvatore Murgia, which passed away few years ago, for his smile and his words of support. vi Vita July 1996 ........................................................High School Diploma, ........................................................................Liceo Classico “De Castro”, Oristano, Italy October 2003 ..................................................Laurea in Biotechnology, Bologna ........................................................................University, Bologna, Italy October to March 2004 ..................................Graduate visitor at the Virology Laboratory, .................................... Department of Veterinary Public Health and ....................................Animal Pathology, University of Bologna, ....................................Bologna, Italy March 2003 to July 2004 ...............................Visiting Scholar at Food animal Health ....................................Research Program, OARDC/OSU July 2004 to February 2005 ...........................Contract at Food animal Health ....................................Research Program, OARDC/OSU April 2005 to November 2005 .......................Contract at the Istituto Zooprofilattico ....................................Sperimentale Della Lombardia e Dell’Emilia ....................................Romagna, Brescia, Italy May 2006 to present ......................................Graduate Research Associate, Veterinary Preventive Medicine, The Ohio State University vii Publications 1. Tibor Farkas, Brittney Fey, Edwin Hargitt III, Mark Parcells, Brian Ladman, Maria Murgia, Yehia Saif. Detection of novel picornaviruses in chickens and turkeys. Virus Gene, accepted for publication November 2011. 2. Abdul Rauf, Mahesh Khatri, Maria V. Murgia, Kwonil Jung and Yehia M. Saif. Differential modulation of cytokine, chemokine and Toll like receptor expression in chickens infected with classical and variant infectious bursal disease virus. Veterinary Research (2011), 42(1):85. 3. Rauf A., Khatri M., Murgia M.V., Saif Y.M. Expression of perforin-granzyme pathway genes in the bursa of infectious bursal disease virus-infected chickens. Dev Comp Immunol (2011), 35(5): 620-627. 4. Tang, Y., Murgia, M.V., Ward, L., Saif, Y. M. Pathogenicity of turkey astroviruses in turkey embryos and poults. Avian Diseases (2006), 50(4): 526- 531. 5. Tang, Y., Murgia, M.V., Saif, Y. M. Molecular characterization of the capsid gene of two serotypes of turkey astroviruses. Avian Diseases (2005), 49(4): 514- 519. Fields of Study Major Field: Veterinary Preventive Medicine viii Table of Contents STUDIES ON TURKEY PARVOVIRUSES .................................................................. 1 DISSERTATION .............................................................................................................. 1 Abstract .............................................................................................................................. ii Acknowledgments ............................................................................................................. v Vita ................................................................................................................................... vii Publications .................................................................................................................... viii Fields of Study ................................................................................................................ viii Table of Contents ............................................................................................................. ix List of Tables ................................................................................................................... xii List of Figures ................................................................................................................. xiii Chapter 1: Literature Review of Animal Autonomously Replicating Parvoviruses .. 1 1.1 Etiology ................................................................................................................ 1 1.2 Epidemiology ...................................................................................................
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