Graduate Student Research Symposium & Schofield Lecture ...€¦ · Graduate Student Research Symposium 2013 2 Introduction The tenth annual Ontario Veterinary College Graduate Student

Graduate Student Research Symposium 2013

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Graduate Student Research Symposium & Schofield Lecture

Conference Proceedings

November 13, 2013


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Introduction

The tenth annual Ontario Veterinary College Graduate Student Research Symposium provides an opportunity for graduate students at the OVC to come

together and celebrate their accomplishments. This year, we have the opportunity to showcase a wide variety of cutting-edge research here at the OVC. We hope you enjoy the presentations that OVC graduate students have

prepared with the support of their advisors and colleagues.

Table of Contents

Schedule.................................................................................................... 3 Keynote Speaker: ..................................................................................... 3 Poster Display Listing................................................................................ 4 Oral Presentation Schedule...................................................................... 6 Abstracts: Oral Presentations................................................................... 8 Abstracts: Poster Presentations............................................................... 21 Acknowledgements................................................................................... 45 Graduate Studies at OVC.......................................................................... 48 Speaker Index............................................................................................ 49


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Graduate Student Research Symposium Schedule

Time Event Location

10:00 am – 11:30 am Posters staffed by graduate students OVC Lifetime Learning Centre

LLC 1707 B & C (Cafeteria)

11:30 am – 12:30 pm Lunch Learning Commons

1:00 pm – 2:00 pm Oral presentations: Session I OVC LLC 1713 and 1715

2:00 pm - 2:10 pm Break

2:10 pm - 3:10 pm Oral presentations: Session II OVC LLC 1713 and 1715

3:10 pm - 3:20 pm Break

3:20 pm – 4:20 pm Oral presentations: Session III OVC LLC 1713 and 1715

4:30 pm – 5:30 pm Schofield Lecture OVC LLC 1714

5:30 pm – 6:30 pm Reception and Awards Ceremony OVC LLC 1707 A (Cafeteria)

Schofield Lecture

“Influenza at the animal-human interface” Dr. Ian York is the team leader for the Molecular Virology and Vaccines Team in the Immunology and Pathogenesis Branch, Influenza Division, Centers for Disease Control and Prevention in Atlanta

York’s current research is focused on antibody-based and cellular immunity to influenza viruses, links between immunity and viral evolution and pathogenicity, and evaluating and improving influenza vaccination. A Guelph graduate with degrees in veterinary medicine and immunology from OVC, York completed a PhD in virology at McMaster University as well as post-doctoral research fellowships at Harvard and the University of Massachusetts Medical School.

York’s talk will provide an overview of influenza evolution and epidemiology, focusing on connections between animal viruses and human disease. He will also discuss ongoing research in his lab and at CDC in general.


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Poster Displays – Room 1707 B & C POSTER SESSION WITH GRADUATE STUDENTS, ROOM 1707 - 10:00 – 11:30

1 Diel de Amorim, M DVSc Proteomic Analysis of the Uterine Flush Fluid from Mares with Endometritis

2 Floras, A DVSc N-terminal pro-C-natriuretic peptide and cytokine kinetics in dogs with endotoxemia

3 Ho, K DVSc Assessing platelet function in clinically healthy cats in response to commonly prescribed anti-platelet medications using three platelet function tests.

4 Sunohara-Neilson, J DVSc Development of an Indirect ELISA for detecting serum antibodies to Leporid herpesvirus 4

using a novel recombinant glycoprotein G* 5 Araujo, C MBS The Regulation of Cardiac CapZ Expression Under Models of Cardiomyopathy Stress

6 Sargent, R MPH Development of an evaluation framework for a multi-faceted and collaborative health promotion strategy at a public health unit

7 Garland, S MPH Summarizing Human Burden of Illness for Extended-Spectrum β-Lactamase-Producing Escherichia coli

8 Fournier, A MPH A scoping review and thematic analysis of the social and economic considerations of the role of wildlife in the transmission of pathogenic bacteria to the food chain

9 Quist, C MPH Hamilton's smoke-free parks and recreation areas by-law: Understanding awareness, support and behaviour change

10 Harding, S MPH Participatory Epidemiology: a Training Workshop 11 Wells, V MPH Antimicrobial Use in Canadian Animals and the Population Correction Unit 12 Swirski, A MSc Temporal trends in Giardia contamination in the Grand River in Waterloo, Ontario (2005-2011)

13 Reilly, K MPH Assessing the viability and impact of hybrid poultry on household livelihoods in central Lao PDR

14 Bishop-Williams, K MSc Mapping heat stress conditions for dairy cattle in southern Ontario- A common geographic

pattern from 2010-2012 15 Gallienne, J MSc The role of PRKAR1a in Canine Osteosarcoma 16 Kumagai, M MSc Development of a Transdermal Delivery System for a Topical NSAID, Meloxicam 17 MacMillan, B MSc Programmed death ligand 2 gene regulation in the context of lentiviral infection 18 Parr, J MSc Common confounders of dietary elimination trials contain the antigens soy, pork, and beef

19 Peacock, H MSc Pericyte coverage is synchronized with vessel outgrowth during tail regeneration in the leopard gecko (Eublepharis macularius)

20 Ternamian, C MSc Targeting Acute Lymphoblastic Leukemia with Oncolytic Virotherapy 21 Syed, Z MSc Oncolytic immunotherapy for the treatment of high-grade glioma

22 Masson, S MSc Canine mast cell tumour (MCT) cells: A model for studying human cancers driven by dysregulation of KIT and VEGFR2

23 Dynes, J MSc The Effects of Low-Dose Metronomic Chemotherapy Administration on Ovarian Function and Female Fertility

24 Remillard, L MSc Exploring the Impact of Pet-Loss on Pet Owners and Implications for Veterinarian-Client Communication

25 Ho, N PhD Metabolic modulators as chemotherapeutic sensitizers for colorectal cancer cells 26 Bujold, A PhD The effect of growth conditions on the expression of Actinobacillus suis adhesin genes

27 Tatone, E PhD Evaluation of a handheld device for the detection of β-hydroxybutyrate pre-calving in dairy cattle

28 Ferris, J PhD Cleavage and blastocyst rates, and sex ratio are altered by oocyte exposure to bisphenol A in Bos taurus

29 Gilchrist, G PhD Telomere and Telomerase Analysis in Bovine Preimplantation Embryos


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30 McCarty, C PhD Effect of Hoof Orientation and Ballast on Acceleration and Vibration in the Hoof and Distal Forelimb Following Simulated Impacts Ex Vivo

31 Pinelli, C PhD High IL-1beta expression in peritumoural stromal cells is associated with more aggressive tumours in an inflammatory mouse model of prostate cancer

32 Perkel, K PhD Understanding metabolomic differences between bovine embryos of different developmental competence

33 Pieper, L PhD Exploring Strategies for Johne’s Disease Prevention in Organic Dairy Herds in Ontario, Canada 34 Quach, A PhD A cytogenetic study of young breeding boars in Canada

35 Slifierz, M PhD Zinc-resistance gene czrC identified in methicillin-resistant Staphylococcus hyicus isolated from pigs with exudative epidermitis

36 Tessier, L PhD Isolation, immunophenotyping and lymphocyte suppressive properties of equine CB-MSC 37 Russell, S PhD Characterization of PIWI-like proteins in early bovine embryogenesis

38 Wong, V PhD Anti-chemotactic effects of canine protein C are dependent on the endothelial protein C receptor (ECPR) expressed in canine neutrophils

39 Russell, K PhD Platelet lysate as alternative to fetal bovine serum in equine mesenchymal stromal cell culture 40 Tscherner, A PhD Variation in miR-34 family expression in bovine gametes and early embryos

41 Tsimakouridze E PhD Diurnal gene expression differences as biomarkers of heart disease in a pressure overload-induced cardiac hypertrophy murine model

42 Gilbert, E PhD Ependymal cells express neural stem/progenitor cell markers during spinal cord regeneration in the leopard gecko (Eublepharis macularius)

43 Bondo, K PhD The Potential for Raccoons to Mechanically Transmit Salmonella

44 Yu, D PhD Tissue tropism and transduction efficiency of adeno-associated virus (AAV) vectors pseudotyped with AAV capsids isolated from domestic animal sources

45 Oluwole, O PhD Testis specific protein Y encoded (TSPY) copy number in various bull tissues

46 Stojsin, A PhD Ratio of serum and follicular fluid anti-mullerian hormone as a predictor of patient embryo utilization and pregnancy


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Oral Presentation Schedule- Room 1713

Time Presenter Topic

Degree Program of Presenters: MSc

13:00 James Ackford Optimizing Foreign Gene Expression in Recombinant Fowl Adenovirus 9 (FAdV-9) Vectors

13:15 Lesley Berghuis Stimulating innate immunity in feedlot cattle: Strategies to induce antimicrobial peptide gene expression

13:30 Jue Tang Estimation of the national sheep scrapie prevalence in Canada via abattoir surveillance

13:45 Samanta Russell The effects of 3TSR and combinational metronomic chemotherapy on epithelial ovarian cancer

Break

Degree Program of Presenters: PhD and DVSc

14:10 Cynthia Weijs Health professionals personal use of facebook: impacts on client impressions of credibility

14:25 Andreia Arruda Spatial and temporal characteristics of porcine reproductive and respiratory syndrome in swine herds from the Niagara

region of Ontario

14:40 Monica Baquero Early effects of bovine γδ T lymphocyte subsets on

macrophages infected with Mycobacterium avium subsp. paratuberculosis infection

14:55 Susan Bland Kidney injury molecule-1: a potential urinary biomarker in cats

Break

Degree Program of Presenters: PhD

15:20 Li Deng Fowl adenovirus 9 ORF1 is important in virus replication in vivo but not in vitro and is able to up-regulate the induction of type I

interferons

15:35 Wendy Pons A systematic review of waterborne disease outbreaks

associated with small drinking water systems in Canada and the United States

15:50 Tara Roberts Risk factors for antimicrobial resistance of Escherichia coli

isolates from Ontario broiler chicken flocks at chick placement: A comparison of three production system types

16:05 Steven Roche The Focus Farm approach to on-farm change for Johne’s

disease prevention and control: Perspectives from the Ontario dairy-cattle industry


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Oral Presentation Schedule- Room 1715

Time Presenter Topic

Degree Program of Presenters: MSc and DVSc

13:00 Alim Nazarali The role of methicillin resistant Staphylococcus pseudintermedius

colonization as a risk factor for development of surgical site infections in dogs undergoing TPLO

13:15 Shawn MacKenzie Accuracy of image guided percutaneous injection of the intervertebral disc in dogs – comparison of three modalities

13:30 Aitor Gallastegui Autonomic Innervation Of Upper and Lower Equine Airways; Anatomical identification and location

13:45 Lynn Williams Frozen and cultured equine umbilical cord blood MSC are equally lymphocyte suppressive in vitro

Break


14:10 Clemence Nash Dairy experts’ thoughts on assessing dairy cattle welfare on Canadian farms – A Delphi survey approach

14:25 Lindsay Bergeron The effects of relaxin peptide administration on rat cerebral vasculature: implications for treatment of ischemic stroke

14:40 Kayla Price Experimental manipulation of the poultry environment to affect the efficacy of live Eimeria vaccination in conventionally reared pullets

14:55 Neda Barjesteh Toll-like receptor ligands induce antiviral responses in chicken macrophages

Break


15:20 Kathy Zurbrigg Comparison of gross and histologic heart lesions and heart weights between market hogs that died in transit to the abattoir and control

hearts from the processing line

15:35 Marlene Paibomesai

The Role of DNA Methylation on Cytokine Production of Immune Response Biased Dairy Cattle

15:50 Bryan Griffin Downregulation of cell surface major histocompatibility complex

class I expression is mediated by the leftmost transcription unit of fowl adenovirus type 9

16:05 Faisal Alibhai Short Term Diurnal Rhythm Disruption Post Myocardial Infarction

Alters Early Inflammatory Responses and Worsens Long Term Outcome


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Oral Presentations Abstracts

Graduate Student Research Symposium


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Optimizing Foreign Gene Expression in Recombinant Fowl Adenovirus 9 (FAdV-9) Vectors James Ackford, Peter Krell and Éva Nagy Department of Pathobiology Fowl adenoviruses (FAdVs) are ubiquitous poultry pathogens with large DNA genomes that are capable of accommodating transgenes. Research in our laboratory aims to develop recombinant FAdVs for use in vaccine delivery and gene therapy. Previously, open reading frames at the left end of the genome of a non-pathogenic FAdV-9 strain were studied, identifying a 2.4kb non-essential region that can be used to generate recombinant FAdVs (recFAdVs). However, while experiments have demonstrated that antibodies (Ab) can be produced by intramuscular vaccination of chickens with recFAdVs, optimizing the host immune response generated by these recFAdVs remains a priority. One way to improve Ab production induced by the recFAdVs is to increase foreign gene expression and protein production by optimizing the expression cassette. I am testing various high expression promoters and enhancer elements to improve transgene expression in recFAdVs both in vitro and in vivo. Expression constructs were cloned using the vector pCI-Neo. Five promoters are studied: the immediate early cytomegalovirus (CMV) promoter, the artificial chicken β-actin/CMV early enhancer (CAG) promoter, the human elongation factor 1α (EF-1α), the chicken β-actin promoter, and fowlpox derived L2R promoter. Promoter activity is compared by measuring enhanced green fluorescent protein (EGFP) of transfected chicken liver (CH-SAH) cells at 12, 24, 36, and 48 hour post-transfection. The data suggest that the CMV and CAG promoters function optimally in CH-SAH cells. Future research will compare promoter expression levels in recFAdVs and in other avian cells.

Stimulating innate immunity in feedlot cattle: Strategies to induce antimicrobial peptide gene expression Lesley Berghuis, Jodi Bierworth, Mary Ellen Clark, Shayan Sharif Neil Karrow, Jeff Caswell Department of Pathobiology Bovine respiratory disease (BRD) is an economically devastating complex of bacterial and viral infections that greatly affects the beef industry across North America. Studies have shown that viral infections such as BVDV and glucocorticoid production in stressed calves inhibit the induction of tracheal antimicrobial peptide (TAP), a cationic β-defensin that has direct microbicidal effects within the respiratory tract. Lipopolysaccharide is known to stimulate TAP gene expression, probably via the TLR4–NF-κB signal transduction pathway, but the maximum effect is only observed after 16 hours of stimulation. This study investigated other agonists of TAP gene expression. PCR analysis of bovine tracheal tissue, lung tissue and unstimulated tracheal epithelial cells showed common mRNA expression for TLRs 1, 2, 3, 4, 5, 6 and 9. In addition to these TLRs, unstimulated epithelial cells showed mRNA expression for IL-17A receptor. Following these findings, tracheal epithelial cells were stimulated with pro-inflammatory cytokines such as IL-17A or IFN-α and agonists for TLRS 1,2,5,6 and 9. Quantitative RT-PCR analysis showed that TLR2 agonists−Pam3csk4 (TLR1/2) and FSL-1 (TLR2/6) −significantly induced TAP gene expression after only 8 hours of stimulation. IL-17A, flagellin (TLR5), and IFN-α also had stimulatory effects after 16 hours of stimulation, but little or no response was found with CpG ODN (TLR9) or lipoteichoic acid (TLR2). Therefore, certain TLR2 agonists may be of value to stimulate innate immunity in immunosuppressed feedlot cattle.


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Estimation of the national sheep scrapie prevalence in Canada via abattoir surveil lance

Jue Tang, Olaf Berke, Paula Menzies

Department of Population Medicine Classical scrapie is a naturally occurring fatal neurodegenerative disease of sheep and goats, causing great economic loss to farmers. In order to inform a future scrapie eradication program for Canada a national prevalence study has been conducted by the Canadian Food Inspection Agency (CFIA) during 2010 to 2012. Abattoir surveillance in the 10 Canadian provinces sampled sheep obex and lymph node tissues. Serial diagnostic testing was based on the Bio-Rad ELISA to screen for cases in combination with parallel testing using western blot (WB) and immunohistochemistry (IHC) to confirm cases. Prevalence estimates and respective confidence intervals, including the shrinkage estimate and Wilson score interval, were compared. Both stratified and non-stratified analyses by province were conducted and compared. Seven cases of classical scrapie were detected among 11,716 traceable sheep out of a total of 12,367 sampled sheep. The national prevalence is estimated at a level of 0.06% with a 95% Wilson confidence interval ranging from 0.0289% to 0.1233%. In conclusion, classical scrapie among sheep in Canada is rare and at a level of about 1 scrapie case per 1,700 sheep. This prevalence level is in the range of reports from other countries. Stratification by province was not effective for confidence interval estimation; therefore non-stratified results are reported here.

The effects of 3TSR and combinational metronomic chemotherapy on epithelial ovarian cancer Samantha Russell , Jack Lawler, J im Petrik Department of Biomedical Sciences Epithelial ovarian cancer (EOC) is the most lethal gynecological cancer effecting women today. Due to the late onset of vague symptoms and a lack of reliable screening techniques, EOC is usually diagnosed in its advanced stages where the five-year survival rate is approximately 20%. Angiogenesis is the formation of new blood vessels from pre-existing vasculature and is believed to play an important role in the growth and metastasis of EOC. Thrombospondin-1 (TSP-1) is a potent antiangiogenic extracellular matrix glycoprotein produced endogenously. We have previously shown that 3TSR, a peptide containing the three type-1 repeat regions of TSP-1, can normalize tumor vasculature, increase tumor perfusion, and increase chemotherapy drug uptake in EOC solid tumors. Due to blood vessel normalization and increased tissue perfusion, we hypothesize that 3TSR will increase tissue uptake of chemotherapy drugs and synergize with metronomic chemotherapy delivery to induce regression of advanced stage disease. We treated our orthotopic, syngeneic mouse model with 3TSR alone and in combination with chemotherapy drugs administered with a maximum tolerated dose (MTD) and metronomic (MET) schedule. 3TSR combined with metronomic delivery of carboplatin and paclitaxel induced significant tumor regression. Combination of 3TSR with chemotherapy drug treatment increased survival compared to untreated mice, or those treated with the drugs individually. The results from this study suggest that 3TSR in combination with metronomic chemotherapy has a significant effect on tumor burden and may be used as a clinical treatment for EOC in the future.


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Health professionals personal use of facebook: impacts on cl ient impressions of credibil ity Cynthia Weijs, Jason Coe, Shannon Majowicz, Serge Desmarais, Andria Jones-Bitton Department of Population Medicine Facebook is nearly ubiquitous for both personal and business communication. As people increasingly seek health information online, opportunities for collision between private and professional lives have increased substantially. At present, there is no universally accepted etiquette for sharing content online. As a result health professionals need to be aware of the impression that content posted on social media profiles may make on clients to avoid reputation damage to themselves, their employer and the profession. Previous research shows that approximately 20% of early-career veterinarians do post content that could be considered “unprofessional”. However, “unprofessional” is not precisely defined, and perceptions of what is “unprofessional” may be evolving along with a general relaxation of social mores in society. Components of credibility – competence, trust and caring -- are considered the foundation of health professional-client relationships. We expected that “unprofessional” content would be recognized as such and result in lower ratings of credibility by members of the general public. Twelve Facebook profiles were developed from all combinations of gender (male/female), profession (veterinarian /physician /public health practitioner) and profile content (unprofessional/neutral). Members of the public participated in an online survey about first impressions of individuals based on their personal Facebook profile. After reviewing a randomly assigned Facebook profile, participants rated the individual on various personality characteristics including credibility (competence, trustworthiness and caring). Final results will be presented. This study will provide a better understanding of the impact that unprofessional content communicated via personal Facebook profiles may have on peoples’ impressions of health professionals.

Spatial and temporal characteristics of porcine reproductive and respiratory syndrome in swine herds from the Niagara region of Ontario. Andreia Goncalves Arruda, Zvonimir Poljak, Robert Friendship, Jane Carpenter, Karen Hand Department of Population Medicine The Niagara region of Ontario has been part of a porcine reproductive and respiratory syndrome (PRRS) area regional control and elimination program from 2010 to present. During this three-year period, an increase in the number of herds becoming positive was noticed during a few months of 2012. The objectives of this study were to investigate whether spatial dependence would change based on phase of the control program and to describe how PRRS status varied over the years. Seventy-five herds were enrolled on a voluntary basis and information on demographics, location and PRRS status was collected. The presence of clustering, clusters and temporal trends were investigated. The mean total number of animals per herd was 1,451 (75-4,500). Clustering analysis for the months of May 2011, June 2012 and August 2013 showed presence of spatial dependence in June 2012, but not at the other two time points. A risk map showed the central eastern region of Niagara to be at a higher risk for being positive during that time. Clusters were not detected at any of the three points in time. Herd level prevalence of PRRS in swine herds has been decreasing over time, with a slight increase from February to June of 2012. In conclusion, the prevalence of PRRS in swine herds in this geographical region appears to be decreasing over time, and the data suggests proximity to other herds might be a risk factor during unexpected incursion of PRRS virus in part of the study population.


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Early effects of bovine γδ T lymphocyte subsets on macrophages infected with Mycobacterium avium subsp. paratuberculosis infection

Monica Baquero, Brandon Plattner

Department of Pathobiology

Following initial Mycobacterium avium subspecies paratuberculosis (Map) infection, some calves apparently successfully clear the pathogen while others become persistently infected. The importance of γδ T lymphocytes during early anti-mycobacterial immunity is recognized; however in bovine paratuberculosis the mechanisms by which the distinct subsets (WC1+ and WC1-) of γδ T lymphocytes are involved in this process remain unclear. The main objective of the research is to understand the mechanisms whereby γδ T lymphocyte subsets contribute to initial clearance of Map or contribute to establishment of persistent infection and progression of disease. The first phase of the project seeks to understand how early γδ T lymphocyte subsets responses mediate macrophage function during Map infection. To achieve this objective monocyte-derived macrophages co-cultured in direct contact or in a transwell system with WC1+ and WC1- γδ T lymphocyte subsets, were infected with Map at an MOI of 10:1. After 24, 48 and 72 hours the expression of CD25 on γδ T lymphocytes, the expression of MHC-I and MHC-II on monocyte derived macrophages and Map viability were analyzed with flow cytometry. IL-17 was quantified using ELISA and NO was measured using the Griess reaction. Our preliminary data shows that activation and cytokine secretion by bovine WC1+ and WC1- γδ T lymphocytes in these co-culture systems are distinct, and this may modulate macrophage function (Map killing) during the first three days after infection with live Map. The effects of both γδ T lymphocyte subsets are distinct if they are in close contact with infected monocyte-derived macrophages.

Kidney injury molecule-1: A potential urinary biomarker in cats Susan Bland, Olivier Cote, Dorothee Bienzle Department of Pathobiology Kidney injury molecule-1 (KIM-1) is a transmembrane protein with extracellular immunoglobulin and mucin domains, and a variable length cytosolic domain. KIM-1 is normally undetectable in urine, but the ectodomain is shed from proximal renal tubular cells into urine with acute kidney disease in rats and humans. The objectives of this study were to determine the feline KIM-1 sequence, to assess expression of the protein in different regions of the kidney, and to determine its presence in urine samples from healthy and diseased cats. Primers to conserved regions of KIM-1 were applied to feline kidney mRNA that had been reverse transcribed to cDNA. Resulting amplicons were sequenced, which identified three isoforms of KIM-1 of 705, 810 and 894bp. The translated full-length feline protein was 81.8, 40.6 and 38.3% similar to canine, human and rat KIM-1 respectively, and contained similar functional domains as in other species. Subsequently the entire gene (approximately 22, 600bp) was amplified and the contiguous sequence assembled. KIM-1 localized to chromosome A1 in the feline genome. Immunohistochemical staining for KIM-1 identified that the protein was expressed almost exclusively in the S3 segment of proximal renal tubule cells in cats with acute kidney disease, but not in cats lacking kidney disease. Measurement of KIM -1 in urine with a rat immunoassay showed that cats with no (n=6) or chronic (n=9) kidney disease lacked detectable KIM-1 in urine, while samples from cats with acute kidney disease (n=5) has positive immunoassay results. These results suggest that 1) urine KIM-1 measurement may be a very sensitive and specific indicator of acute proximal tubule kidney injury in cats; and 2) available immunoassays may be suitable for clinical application.


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Fowl adenovirus 9 ORF1 is important in virus replication in vivo but not in vitro and is able to up-regulate the induction of type I interferons

Li Deng, Peter Krell , Ray Lu, Éva Nagy

Department of Pathobiology Fowl adenoviruses (FAdVs), as non-human adenoviruses, are being considered as potential vaccine vector alternatives to human adenoviruses. Particularly, the work in our laboratory has demonstrated that a mutant deletion virus FAdV-9Δ4 is a good vector candidate for recombinant poultry vaccines. ORF1, a deoxyuridine triphosphatase (dUTPase) homologue, is one of the genes deleted in FAdV-9Δ4, and it is hypothesized to be important in virus replication in vivo. However, the actual function of ORF1 is unknown. Our aim is to characterize the molecular features of ORF1 and to explore its functions. In the present study, it was found that the transcription of ORF1 started at 2 hours post-infection (hpi) and continued until the late phase; the expression of ORF1 protein started at 6 hpi and it was expressed until 24 hpi. In addition, localization studies demonstrated that ORF1 was localized to both cytoplasm and nucleus. In order to investigate the functions, ORF1 knockout virus, FAdV-9ΔORF1, was generated through site-directed mutagenesis and homologous recombination. The in vitro data showed that FAdV-9ΔORF1 possessed similar in vitro characteristics to wtFAdV-9, in terms of virus titers and replication curves. However, the in vivo study showed that FAdV-9ΔORF1 replicated less efficiently compared to wtFAdV-9, evidenced by virus titres in feces and viral genome copy number in tissues of infected chickens. Moreover, it was shown that FAdV-9ΔORF1 induced significantly lower interferons α and β at 12 hpi, compared to wtFAdV-9, suggesting that ORF1 up-regulates the induction of type I interferons.

A systematic review of waterborne disease outbreaks associated with small drinking water systems in Canada and the United States Wendy Pons, Andrew Papadopoulous, Scott McEwen, Andria Jones-Bitton, Ian Young, Katarina Pintar Department of Population Medicine

Small drinking water systems (SDWS) provide water to 16% of Canada’s population, including permanent residents in rural areas and transient populations of tourists and travellers. The term “SDWS” encompasses a variety of water systems and a standard definition for SDWS in North America does not exist. Previous outbreak summaries have shown that in both Canada and the United States, roughly half of the waterborne disease outbreaks occur in SDWS. Outbreak investigations can improve our knowledge about key water system deficiencies so that waterborne outbreaks can be more successfully prevented but they must be detailed enough to ensure the information is useful. This study used a systematic review to identify and analyze microbiological disease outbreak reports involving SDWS throughout Canada and the United States from 1970 to 2013. The review objectives were to identify the leading causes of waterborne disease outbreaks in SDWS and to assess the variety of terminology used to describe SDWS in these reports. This research found common themes that perpetuate through waterborne outbreaks in SDWS and has indicated areas of concern in reporting practices for SDWS if the data is to be useful for decision makers and operators.


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Risk factors for antimicrobial resistance of Escherichia coli isolates from Ontario broiler chicken flocks at chick placement: A comparison of three production system types Tara Roberts, Scot McEwen, Richard Reid-Smith, Jan Sargeant, Agnes Agunos, David Léger, Michelle Guerin Department of Population Medicine Antimicrobial use in chickens is thought to be a risk factor for the development of antimicrobial resistant bacteria that can infect humans via the food chain. Our objective was to determine risk factors for resistance of E. coli isolates obtained at chick placement, with an emphasis on identifying differences between production system types. Seventy-four conventional, thirty-four antimicrobial-free, and seven organically-raised flocks distributed throughout Ontario were enrolled. For each flock, two pooled dust samples (feeders/drinkers, and floors/walls/fans) were collected immediately before chicks arrived from the hatchery, and six pooled swabs of meconium from chick pads representing 300 birds per flock were collected upon arrival. A questionnaire gathered information on hatchery and barn-level factors, including antimicrobial use, vaccination, biosecurity, and management. E. coli was isolated from the majority of flocks. Isolates were tested for resistance to 15 antimicrobials. Differences between production types were identified only for trimethoprim-sulfa; controlling for clustering at the flock and producer levels, for environmental samples, the odds of resistance were lower for isolates from antimicrobial-free flocks compared to conventional flocks (P = 0.031). For chick pad samples, hatchery company, use of vaccines at the hatchery and age of breeder flocks were associated with resistance of E. coli isolates. For environmental samples, all-in-all-out practices, and disinfecting the barn before chick placement were associated with resistance. This study has shown that there were few differences between production types and that factors associated with resistance of E. coli isolates at placement differed between sample types and varied depending on antimicrobial agent.

The Focus Farm approach to on-farm change for Johne’s disease prevention and control: Perspectives from the Ontario dairy-cattle industry Steven Roche, David Kelton, Andria Jones Bitton, Michael Meehan, Michael Von Massow Department of Population Medicine Our goals were to successfully implement and rigorously evaluate the efficacy of a knowledge transfer (KT) program within the Ontario (ON) dairy-cattle industry, aimed at accelerating the uptake of on-farm management practices that are effective for prevention/control of Johne’s disease (JD). ON Focus Farms (ONFF), a learner-centered process, incorporates adult learning theory within an action research/experiential-learning framework to facilitate behavioural change. Producers engage in 4 full-day sessions where small groups discuss/view JD through a variety of lenses (e.g. veterinarian, consumer), and using many techniques (e.g. farm tours, risk assessments, round-table discussions). Pre-post surveys assessed changes in producer perception, attitudes and knowledge, while pre-post risk assessments evaluated JD and on-farm management. ONFF was initiated in 2010, with more than 200 producers participating across 8 regions of the province. Additionally, 80% of participating producers have reported making changes to their on-farm management as a result of ONFF. On average, ONFF respondents exhibited a moderate-good level of JD knowledge (76.5%) and a moderate-low risk of on-farm JD transmission (116/300) at the pre-survey. Average scores among control respondents did not significantly differ from ONFF respondents. At the post-survey, ONFF respondents’ average level of JD knowledge had significantly increased (81.2%) and their risk of on-farm JD transmission had significantly decreased (106/300). Average level of knowledge and risk of JD transmission among control respondents did not significantly change. ONFF accelerated the uptake of on-farm management practices to improve biosecurity, calf health and JD prevention/control, and should be considered an effective method for extension in ON.


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The role of methici l l in resistant Staphylococcus pseudintermedius colonization as a risk factor for development of surgical site infections in dogs undergoing TPLO. Al im Nazarali , Ameet Singh, Scott Weese, Noel Moens, Tom Gibson Department of Clinical Studies Tibial plateau leveling osteotomy (TPLO) is one of the most commonly performed surgical techniques to stabilize a cranial cruciate insufficient stifle in dogs. Numerous studies have reported high surgical site infection (SSI) rates for TPLO, and methicillin-resistant S. pseudintermedius has emerged as a leading cause of these infections. The objective of this study was to evaluate the impact of pre-operative MRSP colonization on TPLO SSIs. A prospective, multi-institutional study of dogs undergoing was undertaken. Within 24 hours of admission, samples of the nares, pharynx, rectum and surgical site were obtained for MRSP screening. Active surveillance of all patients was performed 30 days post-operatively and SSIs were documented according to standard definitions. The overall SSI rate was 21/338 (6.2%), with 9/21 (43%) caused by MRSP. Ten of 338 (3%) dogs were colonized with MRSP preoperatively. Two of ten (20%) MRSP colonized dogs developed MRSP SSI, compared to 7/323 (2.2%) dogs that were not colonized (P=0.026). 7/138 (5%) of dogs were colonized with MRSP at time of post-operative recheck, between 4 and 8 weeks after surgery. 4/7 (57%) of these dogs were colonized at time of initial admission to the hospital. The pre-operative MRSP colonization rate was consistent with studies of similar populations, as was the SSI rate and commonness of MRSP SSI. However, while there was a significant association between SSI and MRSP colonization, most MRSP infections developed in dogs that were not identified as colonized pre-operatively. Further study of the epidemiology and pathophysiology of MRSP TPLO SSI is needed.

Accuracy of image guided percutaneous injection of the intervertebral disc in dogs – comparison of three modalit ies

Shawn Mackenzie, Jeff Caswell , Brigitte Brisson, Luis Gaitero, Heather Chalmers

Department of Clinical Studies

Image guided percutaneous approaches to the intervertebral disc have been used for administration of therapeutics and collection of diagnostic samples. An imaging modality which allows very accurate placement of a needle is desirable for obtaining diagnostic samples and for delivering therapeutic agents. This study evaluates the accuracy of the injection of a gelified ethanol therapeutic agent into the intervertebral discs of dog cadavers using 3 different imaging modalities; fluoroscopy, ultrasonography, and computed tomography. Injection of the material into the nucleus pulposus was achieved in 55 of 78 (71%) of intervertebral discs injected. Both CT and fluoroscopy were significantly more successful than US guided injections. There was no significant difference between CT and fluoroscopy. Injection success rates did not differ between different vertebral sites or between dog cadavers of different weights. Forty-nine (63%) of the injection sites had evidence of contamination following injection and 10 of these had contamination of structures within the vertebral canal. Our findings support that CT, fluoroscopy and ultrasound can all be used to inject a gelified ethanol product into the nucleus pulposus of the intervertebral disc in different regions of the spinal cord with moderate to high success rates; however there is a moderately high rate of contamination observed with this agent.


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Autonomic Innervation Of Upper and Lower Equine Airways; Anatomical identif ication and location Aitor Gallastegui, Norman Ducharme, Wayne McDonell , Laurent Viel Department of Clinical Studies The neural networks of the cervical and thoracic autonomic branches to the airways (sympathetic and parasympathetic) are poorly described in horses. The purpose of this study was to anatomically identify and define this network. Nerve branches in the regions of interest were dissected in 16 horses. At the level of the first rib the right vagus nerve connects with the middle cervical sympathetic ganglia (MCG) in 93.75% of the horses, and frequently with the cervico-thoracic sympathetic ganglia (CTG) or with sympathetic branches from the thoracic sympathetic trunk caudal to the CTG in 66% and 43.75% of the horses, respectively. The left vagus nerve only connects with the MCG in 31.25% of the cases, and with other regions of the thoracic sympathetic system in 25% of the cases. The R and L ventral thoracic vagal branches provide an average of 2 to 3 branches to their ipsilateral bronchi. A third small nerve branch appeared to innervate the laryngeal muscles reaching the larynx within the cranial laryngeal nerve originating at the thoracic vagus nerve, or the thoracic sympathetic trunk. This study showed that a) most innervation to the airways in horses is provided by the right vagus nerve, b) sympathetic and parasympathetic nerves intermingle in relation to the MCG and CTG, and lastly c) a novel branch connecting the thoracic vago-sympathetic nerves and the larynx in horses was identified.

Frozen and cultured equine umbil ical cord blood MSC are equally lymphocyte suppressive in vitro Lynn Wil l iams, Judith Koenig, Laurance Tessier, Thomas Koch Department of Clinical Studies Joint injury is responsible for significant loss in the equine industry. These injuries have recently been treated with intra-articular administration of mesenchymal stromal cells (MSC) despite a lack of experimental evidence. Pre-screening of culture expanded MSC has detected lymphocyte suppressive properties using mixed lymphocyte reactions (MLR). Immediately thawed MSC have not been evaluated using MLR. We hypothesize that frozen and cultured equine umbilical cord blood (CB) MSC have equal lymphocyte suppressive effect in vitro CB-MSC, previously isolated and frozen from five different foals were thawed and culture-expanded in in a routine manner. Cryovials of same CB-MSC stock were maintained in liquid nitrogen for simultaneous evaluation. Peripheral blood lymphocytes (PBL) were co-cultured in 96 well plates with allogeneic mitotically arrested PBL and mitotically arrested CB-MSC. The CB-MSC had been either thawed and cultured for 5-7 days or thawed immediately prior to use. BrdU staining, as proxy for cell proliferation was evaluated by flow cytometry. CB-MSC significantly suppressed PBL proliferation compared to control wells without CB-MSC (p<0.0001). The in vitro lymphocyte suppressive properties of equine CB-MSC were confirmed. The similar effect of frozen and cultured CB-MSC suggest cryopreserved cells could be stocked, thawed, and then injected at the time of injury without the need of sophisticated cell culture equipment. Further in vivo study is needed to assess possible in vivo anti-inflammatory effects of such CB-MSC.


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Dairy experts’ thoughts on assessing dairy cattle welfare on Canadian farms – A Delphi survey approach . Clemence Nash, David Kelton, Derek Haley

Department of Population Medicine

The Canadian dairy industry is constantly evolving. More emphasis is being placed on animal welfare within the industry and their Code of Practice for the care and handling of dairy cattle has recently been revised (2009). An increasing amount of research is being conducted on dairy welfare, however it remains unclear how industry experts are utilizing this knowledge and applying it on-farm. This study aims to investigate how key expert groups in the industry perceive animal welfare and to identify areas of agreement and disagreement between these groups. This project will also work towards developing realistic and optimum standards of care for dairy cattle. To accomplish these objectives we are using Delphi methodology: an iterative online survey process. This research is on-going - we will be presenting the results of the first round of surveys for this abstract. This survey included both open and close-ended questions on respondent demographics, preferred types of welfare measures and thoughts on welfare terminology. We have received 104 completed surveys so far. Of these, 26 were completed by dairy producers, 52 by veterinarians, 20 by researchers, 8 by government workers, 26 by extension workers and 8 by other non-specified groups. Experience ranged from 1.5 years to 59 years within dairy. Of these respondents, 83% specified they would use resource based measures to assess welfare, 74% specified they would use management based measures and 100% specified they would use animal based measures. This demonstrates the focus being placed on the animals themselves by dairy experts assessing welfare on-farm.

The effects of relaxin peptide administration on rat cerebral vasculature: implications for treatment of ischemic stroke. Lindsay H. Bergeron, Saad Enouri, Ron Johnson, Brian Wilson, Stephanie Nykamp, Alastair J.S Summerlee Department of Biomedical Science Ischemic stroke occurs when a cerebral vessel is occluded, resulting in tissue deprivation of oxygen and nutrients. Without sufficient reperfusion, the area of severely compromised blood flow, the ‘core’, becomes irreversibly damaged. The area surrounding the core, the ‘penumbra’, is transiently maintained by peripheral blood supply but is at risk of irreversible damage, making it an ideal target for medical intervention. Relaxin peptides, members of the insulin/relaxin superfamily, are potent vasodilators and have demonstrated protective effects in multiple ischemic tissues. We investigated the effects of relaxin peptides on infarct size in a rat stoke model. H2 relaxin, H3 relaxin or saline was injected intravenously into anaesthetized rats thirty-minutes prior to occlusion of the middle cerebral artery. Brains were removed four hours after stroke and coronal sections were stained for infarct size determination. These experiments were repeated with the addition of an intracortical injection of L-NIO, an endothelial nitric oxide synthase inhibitor, in order to determine the mechanism of observed effects. We also studied the effect of relaxin peptides on myogenic reactivity of cerebral vasculature. Pressure myography was used to study cannulated rat middle cerebral arteries treated with H2 or H3 relaxin (including appropriate controls). Relaxin peptides significantly reduced infarct size compared to saline-treated controls. Relaxin-mediated protection was abolished with L-NIO. Relaxin peptides altered myogenic reactivity in rat middle cerebral arteries. Results suggest that relaxin peptides provide protection to the ischemic brain through modifications of cerebral vasculature, possibly improving perfusion status of the penumbra, thus limiting the extent of brain injury.


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Experimental manipulation of the poultry environment to affect the efficacy of l ive Eimeria vaccination in conventionally reared pullets Kayla Price, Michele Guerin, John Barta Department of Pathobiology Live Eimeria vaccination stimulates immunity from the original vaccine oocysts and is enhanced through fecal-oral transmission (“cycling”). Vaccination is used successfully with birds reared on litter because sufficient environmental oocyst cycling takes place post inoculation; for birds reared on wire mesh flooring, low-level oocyst cycling is less efficient and chickens may remain susceptible. Environmental management becomes critical for vaccine success. Two trials evaluated biodegradable cage floor coverage (CFC) of the mesh floor (lasting ~5 weeks) at different percentages to enhance oocyst cycling. Pullets were vaccinated with a low-dose of oocysts at commercial vaccine proportions by oral-gavage (study 1) or spray vaccination (study 2). Birds were reared under commercial conditions from 0 to 6 weeks of age. In the primary study, birds were reared on 0%, 20%, 40% or 60% CFC whereas 0% or 40% CFC was used in the subsequent larger-scale study. At 6 weeks of age birds were challenged with mixed Eimeria species (study 1) or mixed and single species (study 2). Lesion scores, weight gains and oocyst shedding post challenge were assessed Experimental manipulation of the poultry environment to affect the efficacy of live Eimeria vaccination in conventionally reared pullets.

Toll - l ike receptor l igands induce antiviral responses in chicken macrophages Neda Barjesteh, Alexander Ian Vil lanueva, Shahriar Behboudi, Eva Nagy, Shavan Sharif Department of Pathobiology Chicken macrophages express several receptors for recognition of pathogens, including Toll-like receptors (TLRs). TLRs bind to pathogen-associated molecular patterns (PAMPs) leading to activation of macrophages. However, the effects of TLR ligand dose, time of administration and the mechanisms through which TLRs modulate the antiviral responses of chicken macrophages have not been well characterized. The present study was designed to test the hypothesis that treatment of chicken macrophages with TLR ligands reduces avian influenza replication in these cells. Furthermore, we sought to examine the aforementioned factors and determine the mechanisms involved in TLR-mediated antiviral responses of macrophages. Chicken macrophages were treated with TLR2, -3, -4, -7 and -21 ligands: Pam3CSK4, polyI:C, LPS, R848 and CpG respectively, with different doses and at different time points, pre- and post- avian influenza virus (AIV) H4N6 infection. The results revealed that the pre-treatment of macrophages with appropriate doses of Pam3CSK4, LPS and CpG reduced the replication of AIV in chicken macrophages. In addition, the relative expression of some genes was quantified at 3, 8 and 18 hours post-treatment with TLR2, -4 and -21 ligands. Pam3CSK4, LPS and CpG increased the expression of interleukin (IL)-1β, interferon (IFN)-γ, IFN-β and IFN-7. The expression of these genes correlated with the reduction of viral replication in macrophages. In conclusion, two main findings emerged in the current study; the dosage of TLR ligand used and the time of administration of TLR ligands determine the ability of macrophages to control AIV replication. These results shed more light on the process of immunity to AIV in chickens.


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Comparison of gross and histologic heart lesions and heart weights between market hogs that died in transit to the abattoir and control hearts from the processing l ine. Kathy Zurbrigg, Tony van Dreumel, Robert Friendship, Terri O’Sull ivan Department of Population Medicine The study objective was to determine if pre-existing diseases predisposed market hogs to sudden death during transport. Post-mortems on 47 in-transit loss (ITL) hogs from an Ontario abattoir were performed at the Animal Health Laboratory, University of Guelph. Twenty-eight hearts collected from hogs that did not die in transit were collected for comparison. Each heart was weighed and examined blindly by a veterinary pathologist (TVD). ITL hearts were compared to non-ITL hearts. Gross heart lesions were observed in 96% (45/47) of ITL hogs. Left ventricular hypertrophy was observed in 87% (41/47) of ITL cases. Affected hearts also had combinations of subaortic fibrosis, and dilation of the right ventricle, pulmonary artery and aorta. The 45 ITL cases with gross heart lesions had combinations of the following histologic lesions: myocardial cell disarray, medial hyperplasia of intramural coronary arteries and fibrosis and atrophy of myocardial fibres. No lesions were found on gross examination of the 28 control hearts and 29% (8/28) had no histologic lesions. The remaining 20 control hearts (71%) had histologic lesions similar to the ITL cases, although significantly milder (P<.05). The total heart weight of ITL hearts (448.60gm + 68.7SD) was greater than non-ITL hearts (372.80gm + 42.7SD) (P<.05). This was also true for the left ventricle and septum (278.55gm + 46.8SD vs 248.25gm + 27.74SD) and the right ventricle (97.85gm + 23.7SD vs 78.41gm + 15.31SD)(P<.05). The majority of hogs that died in transit to an Ontario abattoir had cardiac abnormalities that were significantly associated with death during transport.

The Role of DNA Methylation on Cytokine Production of Immune Response Biased Dairy Cattle

Marlene Paibomesai and Bonnie Mallard


Epigenetic modifications control gene expression without changes to the germline DNA sequence. These modifications, such as DNA methylation, have been extensively studied in the mouse CD4+ T-cell immune response (IR) and it has been shown that specific DNA methylation sites within the promoter region of IFN-γ and IL-4 have an influence on the overall gene expression and subsequent synthesis of these cytokines. Therefore, the objective of this study was to determine the role of DNA methylation at cytokine promoters in antibody (AMIR) and cell (CMIR) mediated immune response biased cattle. Blood was collected from HighAMIR-LowCMIR (HiAMIR; n=12) and HighCMIR-LowAMIR (HiCMIR; n=11) cows three weeks after calving. Isolated CD4+ T-cells were collected and stimulated for 24 hours with Concanavalin A at which time genomic DNA was collected for subsequent bisulfite pyrosequencing assay. It was previously shown that cells isolated from HiCMIR cows produced more IL-4, IL17A and a tendency to produce more IFN-γ 21 days after calving than CD4+ cells from HiAMIR cows. Preliminary results show DNA methylation change across methylation sites at the IL-4 promoter ranging from 30-60% methylation. Despite this range of methylation across CpG sites little difference was observed between unstimulated and stimulated CD4+ T-cells at specific CpG sites. A verifying population of CD4+ T-cells that were skewed to expressing high IL-4 transcript showed a 20% decrease in methylation across these CpG sites. Therefore, these CpG sites may not be responsible for immune response variation in IL-4 production of CD4+ T-cells from immune response biased dairy cattle.


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Downregulation of cell surface major histocompatibi l i ty complex class I expression is mediated by the leftmost transcription unit of fowl adenovirus type 9 Bryan Griff in and Éva Nagy Department of Pathobiology Adenoviruses (AdVs) are members of the family Adenoviridae and are nonenveloped, icosahedral viruses that infect a wide range of vertebrate hosts. Fowl adenoviruses (FAdVs) that infect birds are in the genus Aviadenovirus. Some FAdVs are associated with inclusion body hepatitis (IBH), characterized by hepatic necrosis and intranuclear inclusion bodies. While mammalian adenoviruses are well-studied, protein functions of the most studied FAdV, FAdV-1, have been demonstrated for only three of the predicted 19 genus-specific genes, ORF1, ORF22, and GAM-1. Although FAdVs do not have an E3 region, the region responsible for MHC-I downregulation in human ADVs, we hypothesized that one or more of the FAdV genus-specific genes down-regulate surface expression of BF2, the MHC-I molecule of the chicken. As determined by flow cytometric analysis with anti-chicken MHC-I, the normally high MHC-I cell surface expression of chicken hepatoma cells (CH-SAH) was reduced to approximately 15% of initial levels upon infection with FAdV-9 at a multiplicity of infection of 5. The left-end transcription unit deletion virus (FAdV-9∆4), in contrast, reduced cell surface MHC-I to approximately 50%. Western blotting of protein lysates of uninfected, parental, and knockout virus-infected CH-SAH with anti-chicken MHC-I antibody revealed that total MHC-I was reduced equally among the viruses. We, therefore, hypothesize that MHC-I is sequestered intracellularly via direct binding with an FAdV-9 protein(s) or through disruption of intracellular transport. Since the epitope recognized by the monoclonal anti-chicken MHC-I antibody was degraded upon para-formaldehyde fixation, cDNA encoding the dominantly expressed MHC-I was cloned into pFLAG-CMV-1 for visualization of MHC-I upon parental and mutant virus-infection via immunofluorescence analysis (IFA).

Short Term Diurnal Rhythm Disruption Post Myocardial Infarction Alters Early Inflammatory Responses and Worsens Long Term Outcome Faisal J. Al ibhai, Elena Tsimakouridze, Glen Pyle, Tami Martino

Department of Biomedical Science

Rationale: Circadian rhythms allow for coordination of animal physiology in a 24hr environment. This is achieved through synchronization with external light cues to produce rhythms that exhibit diurnal (day/night) variation. Following a myocardial infarction (MI) coordination of cellular processes is required to promote effective infarct healing. However, little is known about the role of diurnal rhythms in the pathophysiology post-MI. Hypothesis: Short term diurnal rhythm disruption adversely affects outcome post-MI in mice. Methods and Results: Mice were infarcted by left anterior descending coronary artery ligation between ZT 1-4, randomized to either a normal diurnal or disrupted environment for 5 days then maintained under normal diurnal conditions. Both disrupted-MI and normal-MI mice had similar infarct sizes at day 1 post-MI however, diurnal disruption altered inflammatory cytokine production and immune cell infiltration in the infarcted myocardium over the first 7 days post-MI. These early alterations led to impairments in scar formation and exacerbated cardiac remodeling in disrupted-MI mice by 1 week post-MI as assessed by echocardiography and histology. By 8 weeks post-MI alterations in early cardiac remodeling led to impaired myofilament function and significantly poorer cardiac structure and function as assessed by echocardiography and hemodynamics. Histological analysis revealed significantly greater infarct expansion and left ventricular dilation in mice subjected to short term diurnal rhythm disruption. Conclusions: Rhythm disruption post-MI worsens cardiac remodeling and progression to heart failure. The implications are significant; disrupted diurnal rhythms such as those found in modern intensive care environments adversely affect long term patient outcome.


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Poster Presentations Abstracts

Graduate Student Research Symposium


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Poster 1 Proteomic Analysis of the Uterine Flush Fluid from Mares with Endometrit is Mariana Diel de Amorim, Tracey Chenier, Bette Anne Quinn, El izabeth Scholtz, Cathy Gartley, Keith Betteridge, Tony Hayes Department of Population Medicine Bacterial uterine infections inflict major losses on the equine breeding industry, occurring in 25–60% of mares that fail to become pregnant. Endometrial cytology, culture and biopsy are routinely used to diagnose endometritis in mares. However, many mares fail to conceive after being diagnosed negative for endometritis. Our objective is to identify the major proteins in the uterine flush fluid of mares, and to identify those that change in association with endometritis. Low volume lavage (LVL) followed by endometrial biopsy was performed on 38 mares, including mares in estrus and diestrus with or without endometritis. Endometritis was diagnosed in mares with two or more of five criteria (uterine fluid or excessive edema on ultrasound or history of subfertility; 2 or more neutrophils per high power field on cytology; cloudy LVL; positive culture; and active inflammation on biopsy). Proteins in the supernatant of the LVL were identified by LC-MS/MS. Over 2000 proteins were identified and the 200 most abundant were compared among groups by 2-way ANOVA with SAS 9.3 software. Uterocalin/P19, Secretoglobin1A1 and Secretory phospholipase A2 changed significantly with cycle but not with endometritis. By comparison, Vanin 1, Vanin 2, Vanin 3, Connective tissue growth factor, Stanniocalcin and Annexin A2 were among those that were significantly (p ≤ 0.05) altered in mares with endometritis compared to controls. Further comparisons of these and other members of the uterine proteome are needed to determine which proteins are most useful in the diagnostic or treatment assessment of mares that fail to become pregnant.

Poster 2 N-terminal pro-C-natriuretic peptide and cytokine kinetics in dogs with endotoxemia

Alexandra Floras, Marie Holowaychuk, Dorothee Bienzle, Alexa Bersenas, Shayan Sharif, Tami Harvey, Geoffrey Wood

Department of Clinical Studies

Background: Rapid identification of infection in patients with sepsis enables prompt administration of antibiotics that are essential to improve survival. Serum N-terminal pro-C-natriuretic peptide (NT-proCNP) concentration can differentiate naturally occurring sepsis from non-septic inflammation, however, little is known about serum NT-proCNP concentrations in dogs during the course of sepsis. Objective: The purpose of this study was to determine serum NT-proCNP and cytokine kinetics in dogs with endotoxemia, an established model of canine sepsis. Samples: Eighty canine serum samples. Methods: Eight healthy adult Beagles were randomized to receive Escherichia coli O127:B8 lipopolysaccharide (LPS, 5 ug/kg, IV) or placebo (0.9% NaCl, equal volume, IV) in a randomized crossover study. Serum collected at 0, 1, 2, 4 and 24 hours was stored at -80oC for batch analysis. NT-proCNP was measured by ELISA and 13 cytokines and chemokines by multiplex magnetic bead-based assay. Results: Serum NT-proCNP concentrations did not differ significantly between LPS- and placebo-treated dogs at any time. When comparing serum cytokine concentrations in LPS- to placebo-treated dogs, IL-6, IL-10, TNF-α and KC-like were elevated at 1, 2, and 4 hours; CCL2 was elevated at 1, 2, 4 and 24 hours; and IL-8 and CXCL10 were elevated at 4 hours (p<0.05) in dogs treated with LPS. There were no differences in serum GM-CSF, IFN-γ, IL-2, IL-7, IL-15 or IL-18 between groups of dogs. Conclusions: In dogs, serum NT-proCNP concentration does not distinguish LPS-induced inflammation from health. Serum cytokines and chemokines significantly increased within 1-4 hours of LPS administration in dogs warrant further investigation.


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Poster 3

Assessing platelet function in cl inically healthy cats in response to commonly prescribed anti-platelet medications using three platelet function tests. Kimberly Ho, Anthony Abramms-Ogg, Shauna Blois, Gord Kirby, Lynne O’Sull ivan, Darren Wood Department of Clinical Studies Feline hypertrophic cardiomyopathy (HCM) increases the risk of developing fatal thromboembolism. Cats with HCM are commonly prescribed anti-platelet medications with limited proven efficacy of their use in this species. This study aimed to develop institutional reference intervals for three analyzers which evaluate platelet function by different means (Multiplate® (by impedance), Platelet Function Analyzer-100® (mechanically), and Plateletworks® (by enumeration), and to use these tests to evaluate the effect of commonly prescribed anti-platelet drugs on feline platelets. Atraumatic venipuncture was performed on 60 clinically healthy cats (mean age 5.51yr, range (1-15yr). Platelet function response to ADP and collagen ± arachidonic acid was assessed using the aforementioned tests and reference intervals were generated for each using EP Evaluator software (CI 95%). To evaluate platelet response to anti-platelet drugs, 24 clinically healthy cats were randomly assigned to receive one of 3 treatments on days 1 – 8: 5mg or 20.25mg ASA PO q72h or clopidogrel 18.75mg PO q24h. Additionally, cats receiving ASA also received dual-agent therapy with the addition of clopidogrel 18.75mg PO q24h on days 4-8. Blood samples were acquired on days 1, 4, and 8, to evaluate platelet function. Significant differences in platelet function were found in cats receiving clopidogrel alone, and in cats receiving clopidogrel and ASA at either dose, when platelet function was assessed using Plateletworks (p<0.05). No significant differences in platelet function were found in any treatment group when platelet function was assessed with Multiplate or PFA-100 (p<0.05). In conclusion, Plateletworks demonstrated an effect of clopidogrel therapy on feline platelet function..

Poster 4 Development of an Indirect ELISA for detecting serum antibodies to Leporid herpesvirus 4 using a novel recombinant glycoprotein G Janet Sunohara-Neilson, Éva Nagy, Susy Carman, Patricia V Turner Department of Pathobiology Leporid herpesvirus-4 (LHV4) is a recently identified alphaherpesvirus that infects domestic rabbits (Oryctolagus cuniculus). Viral infection results in nonsuppurative pneumonia with 80% morbidity and 20% mortality in natural infections. Experimentally, serum antibodies are produced 10 days after intranasal inoculation of rabbits. Antibody detection would be very useful in identifying latently infected animals but no diagnostic assays are currently available. To that end, full length glycoprotein G (gG) gene was amplified from genomic DNA of the concentrated virus isolated from a known positive case and cloned into a baculovirus vector. The cloned sequence is 2505 bp and encodes 835 amino acids. The recombinant protein is expressed in insect cells and cell lysates were examined by SDS-PAGE, Western blotting, and ELISA. Positive sera from experimentally infected rabbits detected a series of bands from 45-65 kDa that were interpreted as the glycosylated cleavage products of gG. This pattern has previously been demonstrated for a homologous protein of human herpesvirus-2. The infected insect cell lysate was used as the antigen in an indirect ELISA. Sera were collected at abattoirs from commercial rabbits produced at farms across Ontario to assay for potential viral exposure. Results from this new diagnostic assay agree with those obtained from positive and negative controls and abattoir sera using an in-house virus neutralization assay. This new gG-based indirect ELISA could be used to generate rapid results in outbreak situations or as a routine screening tool to identify latently infected rabbits.


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Poster 5 The Regulation of Cardiac CapZ Expression Under Models of Cardiomyopathy Stress Chris Araujo, Elena Tsimakouridze, Faisal Al ibhai, W. Glen Pyle, Tami A. Martino Department of Biomedical Sciences CapZ is a ubiquitous actin-binding protein which anchors sarcomeric actin to Z-discs in cardiac myocytes. We have previously demonstrated that a modest decrease in CapZ protects hearts against ischaemia-reperfusion injury. Whether CapZ regulation is an endogenous event that occurs with cardiac stress is unknown. BAG3-HSC70, as well as CapZ-interacting protein (CapZIP), alter CapZ which alters actin binding in vitro. The objectives of the current study were to investigate if CapZ regulatory pathways are modulated in heart failure and identify the consequent impact on CapZ. Myocardial samples were taken from dogs in the end stage of natural dilated cardiomyopathy (DCM) and from mice with ischaemic cardiomyopathy (ICM) due to coronary artery ligation. Myofilaments were isolated and subjected to immunoblotting. CapZ protein increased by 118% in canine DCM myofilaments. BAG3 and HSC70, which stabilize CapZ-actin binding, increased by 71% and 58% respectively. CapZIP phosphorylation also promotes CapZ binding in actin, and increased at S179 by 169%, while S108 remained unaffected. Total CapZIP protein levels exhibited no change in canine DCM samples. CapZ in murine ICM myofilaments decreased 27%. CapZIP phosphorylation decreased at S108 by 24%, and myofilament-associated HSC70 decreased by 22%. BAG3, CapZip S179 phosphorylation, and CapZIP levels were unaffected. These data present the novel findings that CapZ is differentially affected in heart failure. These changes involve condition-specific effects on BAG3-HSC70 and CapZIP pathways. A further understanding of CapZ regulation in the heart is integral to the development of CapZ as a therapeutic target in the heart.

Poster 6 Development of an evaluation framework for a multi -faceted and collaborative health promotion strategy at a public health unit Robbyn Sargent, Chris Harold, Meghan Randall , Grace Bermingham Department of Population Medicine The Region of Waterloo Public Health reviewed their sexual health services and found that their teen pregnancy and youth STI rates were higher than the provincial average. This led to the development of a Waterloo Region Sexual Health Youth Strategy that involves many community agencies and includes thirty activities that target youth and their parents. An evaluation framework was developed for the Waterloo Region Sexual Health Youth Strategy, which includes consideration of innovative evaluation techniques and development of evaluation indicators and questions. Evaluation approaches, data collection methods and process, and outcome objectives for each activity were outlined. Activities to be evaluated were prioritized based on importance, expected impact and how developmental they were. This was necessary as human and financial resources are very limited for this evaluation. A timeline for evaluation was defined to allow for most activities to be evaluated. Activities will be evaluated using a variety of techniques, ranging from outcome indicators to focus groups, surveys and community-wide statistics.


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Poster 7 Summarizing Human Burden of I l lness for Extended-Spectrum β -Lactamase-Producing Escherichia coli Sarah Garland, Carolee Carson, Rita Finley, Jane Parmley Department of Population Medicine

Extended-spectrum β-lactamase (ESBL) -production in Escherichia coli (ESBL-EC) confers resistance to penicillins, monobactams and the extended-spectrum cephalosporins, making clinical treatment of ESBL-EC infections challenging. We identified and described the literature reporting human burden of illness (BOI) measures due to infection with ESBL-EC. Scientific literature was identified through the use of a validated search string, screened based on relevance, and pre-defined inclusion/exclusion criterion. Descriptive summaries of the captured data were created for BOI measures with few reports or where cross-study synthesis was not possible. For BOI measures with three or more studies a stochastic simulation model was used to generate a quantitative summary measure of the BOI indicator. Meta-analyses were performed for mortality and antimicrobial use BOI measures. Forty-seven relevant articles were included, with 11 BOI measures identified. There were no Canadian studies identified. A lack of standardization for BOI measures existed across studies, making synthesis challenging. Higher cost, higher mortality, longer length of stay, more treatment failure, inappropriate initial antimicrobial therapy, higher intensive care unit admission, and longer time to adequate treatment were documented for persons with ESBL-EC compared to those with non-ESBL-EC. Sepsis and other BOI outcome measures may or may not be worse for ESBL-EC. Any antimicrobial use, β-lactams (general), cephems, or quinolones may be associated with increased risk of ESBL infection. There are many BOI measures identified in the literature; however, they lack standardization across studies. Infection with ESBL-EC may result in higher burden of illness and worse outcomes for patients, compared to infection with susceptible bacteria.

Poster 8 A scoping review and thematic analysis of the social and economic considerations of the role of wildl ife in the transmission of pathogenic bacteria to the food chain Alexandra Fournier, Ian Young, Judy Greig, Andri jana Rajic, Jeff LeJeune Department of Population Medicine The transmission of pathogenic bacteria to the food chain has caused damaging impacts on the agri-food industry and public health. Many agencies have concentrated management programs on removing the threat of pathogen transmission from wildlife to the food chain. However, current wildlife management practices have faced stakeholder opposition and resistance due to conflicting values and interests, and a better understanding of the social and economic considerations of this issue is needed. A mixed-method scoping review of the literature was conducted to summarize and describe the key research areas and socio-economic aspects of the issue, and to identify key factors for the success of potential risk prevention strategies. A total of 20,036 citations were screened and 141 relevant articles were identified that investigated contextual aspects of this issue. Thirty of these articles were prioritized by two reviewers for thematic analysis. We identified two key thematic areas that were consistent throughout the literature: 1) the role of wildlife in pathogenic bacteria transmission to the food chain as a ‘wicked problem’, which consisted of four sub-themes: 1a) social factors, 1b) governance, 1c) conservation and 1d) economic implications. The second key thematic area identified was: 2) promising approaches to prevent and mitigate these potential risks, which included three sub-themes: 2a) collaborative decision-making approaches, 2b) evidence-informed approaches and 2c) the importance of local context. The integration of this information will play a key role at improving the acceptability and sustainability of future strategies to reduce transmission of pathogenic bacteria between wildlife and food animals.


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Poster 9 Hamilton's smoke-free parks and recreation areas by-law: Understanding awareness, support and behaviour change Crystal Quist, Heidi McGuire, Ryan Kennedy, Stephanie Fi lsinger Department of Population Medicine The objective of this study was to evaluate the support, behaviour change and social norm change associated with smoke-free parks and recreation areas by-law implemented by the City of Hamilton on May 31, 2012. This evaluation employed a mixed-methods approach. Research methods included intercept surveys in city-owned parks and recreation areas with Hamilton residents and visitors, online surveys for recreational and neighbourhood associations, field observations of smoking behaviour in city parks and recreation areas, a focus group interview with tobacco enforcement staff and an audit of cigarette butt litter at selected sites. Intercept surveys were completed with 511 residents 18 years of age and older, who were recreation area users (80% response rate). More than 80% of respondents supported regulating smoking in outdoor recreational areas. In addition, sports and neighbourhood organizations surveyed reported similarly high levels of support. More than half of residents stated that they usually or always witness smoking/smoked themselves in these areas. And almost all of them reported often or always noticing litter caused by cigarette butts. The litter audit had similar findings; although there has been a decrease in the number of cigarette butts within these areas post-by-law. These findings suggest a strong level of support for smoke-free parks and recreation areas. Furthermore, the data illustrates the potential effects of policy on behaviour change and de-normalization of smoking. Therefore, the information obtained from this study provides evidence that outdoor smoke-free policy can support tobacco control efforts in health protection and smoking prevention that are occurring across Canada.

Poster 10 Participatory Epidemiology: a Training Workshop Shannon Harding, Jane Parmley, Morrison Karen Department of Population Medicine Participatory epidemiology (PE) is an adaptive methodology that can provide the contextual knowledge needed in all communities to better understand important disease issues. Recognizing a local interest in the topic, we conducted a training workshop at the Ontario Veterinary College in May 2013. Our goal was to have trainees learn about PE and be able to use participatory tools and techniques. The fieldwork component of the course consisted of a case study which investigated factors influencing enteric disease in Southern Ontario. Semi-structured interviews and focus groups were conducted with public health professionals. A content analysis was then performed on the qualitative results to identify common risk factor themes which included: travel, food handling, farm-to-fork, water, geography, demographics, and behaviours. Risk factor complexities and interviewee sensitivities to certain subject matter were also identified. To evaluate the success of the course, trainees completed pre and post workshop surveys. Results indicated that all respondents agreed that the workshop was successful and that it should be offered again. Key recommendations for future workshops were to have the workshop be longer to accommodate more fieldwork practice and to place a greater emphasis on learning more qualitative data analysis techniques


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Poster 11 Antimicrobial Use in Canadian Animals and the Population Correction Unit Victoria Wells, Carolee Carson, David Leger, Rita Finley Department of Population Medicine Antimicrobial resistance (AMR) is a concern in Canada, and one of the driving risk factors behind it is antimicrobial use (AMU), however antimicrobials are critical for treating infectious diseases. Different measures of AMU can provide different types of information and appropriate denominators are needed to provide the context of population exposure. The population correction unit (PCU) is one denominator for animal AMU data that adjusts the weight of antimicrobials distributed for use in animals (provided by the Canadian Animal Health Institute) by the biomass of the population at risk. Using the quantity of antimicrobials distributed for sale and this quantity adjusted by biomass, we have reported on AMU in animals in Canada from 2006 to 2012. Ionophores, an antimicrobial class considered of low importance to human medicine, and tetracyclines make up a large proportion of national animal AMU. The quantities of AMU vary across the provinces and in some cases there is a different ranking of drug classes. The reasons for these differences are currently unknown, but could be due to variation in the food animal species populations between provinces, differences in disease incidence, or differences in use practices. Using preliminary data, we compared Canada (mg/PCU) to similarly reporting countries within the EU. Canadian animal AMU appears to be generally increasing over time when corrected for biomass, but generally decreasing when measured as volume of drugs distributed for sale. Companion animal use represents 13% of the quantity of antimicrobials distributed. Future work on this project will involve engaging the agricultural industry to validate and improve the PCU denominator.

Poster 12 Temporal trends in Giardia contamination in the Grand River in Waterloo, Ontario (2005-2011). Alexandra Swirski, David Pearl, Andrew Peregrine, Katerina Pintar Department of Population Medicine Giardia lamblia, the etiologic agent of human giardiasis, is the most frequently identified intestinal parasite of people in North America. As part of the Public Health Agency of Canada’s C-EnterNet surveillance program, water samples were collected monthly from the Grand River in Waterloo, Ontario, from 2005 to 2011 and Giardia cyst concentrations were determined. Our study objectives were to describe the temporal pattern, both secular and seasonal, of water contamination with Giardia in the Grand River and to determine if water quality parameters may act as effective indicators for Giardia contamination of the Grand River. Multivariable linear regression models were produced with temporal and water quality parameter data to determine their association with Giardia cyst concentration in the river. Season and year were significant predictors for river contamination with Giardia. Cyst concentrations were significantly higher in the winter and spring compared to the summer and fall, and were significantly higher from 2005-2008 compared to 2009-2011. Nitrite concentration was a significant predictor within the water quality parameter model, and was negatively associated with Giardia contamination, after controlling for the volumetric flow rate of the river water. Interestingly, the seasonal pattern of water contamination found in this study does not match the temporal pattern of human giardiasis cases previously reported in Ontario. These findings are important to help guide our understanding of source water pathogen levels and to guide local decision-making around watershed management and public health protection.


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Poster 13 Assessing the viabil ity and impact of hybrid poultry on household l ivelihoods in central Lao PDR. Kristen Reil ly, Malavanh Cittavong, Anne Drew, Lampheuy Kaensombath, Kham Phommachanh, Blanaid Donnelly, Sonia Fevre

Department of Population Medicine Smallholder poultry are critical for alleviating poverty and food insecurity in developing countries. Education and training initiatives in Lao PDR aim to enhance poultry production and survival, however women in central regions are underserviced despite their prominent role in poultry rearing. Local chickens require few inputs but produce low yields of eggs and meat, whereas hybrid poultry breeds have potential for superior production but require greater investments in feed and vaccines. A pilot study conducted in three rural villages within Vientiane Capital assessed the viability of hybrid poultry for smallholder farmers. Participants (N = 41) received training, a manual, 10 hybrid chicks, 17kg of feed and mentoring visits. Surveys evaluated change over time using descriptive statistics, Chi-Square, Fisher’s Exact and McNemar tests. Local poultry vaccination increased by 40% after the intervention while no changes were observed in poultry sales. More women reported low-literacy than men (P < 0.05) but there were no gender differences in poultry vaccination, product sales or profit reinvestment. The greater availability of meat and eggs was the most commonly reported project benefit (55%) while hybrid poultry mortality (68%) and increased expenses (22%) were the most frequently reported project drawbacks. Results demonstrate that poultry-rearing is a shared household activity and not solely the responsibility of women (48%). Currently, hybrid poultry are not considered to be a viable means of poverty alleviation since farmers are unable to make necessary investments. Further research into household division of labour and community-based management approaches is needed to promote low-investment local poultry production.

Poster 14 Mapping heat stress conditions for dairy cattle in southern Ontario- A common geographic pattern from 2010-2012. Katherine Bishop-Wil l iams, Olaf Berke, David Pearl, David Kelton Department of Population Medicine In southern Ontario, climate change has resulted in an increased occurrence of heat waves, causing heat stress among humans and livestock, with potentially fatal consequences. Heat waves are defined as three consecutive days of temperatures greater than or equal to 32°C. Heat stress is a measure of discomfort relative to temperature and humidity in the ambient air. Maps visualizing the distribution of heat stress can provide information about related health risks and insight for control strategies. Weather data were collected from meteorological stations throughout southern Ontario for dry bulb temperature and dew point temperature. The Dairy Cow Heat Stress Index (HSI), was estimated by averaging the first three days for three heat waves, during 2010-2012. Geostatistical kriging was used to map three-day averages of maximum heat stress over periods involving a heat wave and control periods three weeks prior to and following heat waves. Average HSI for each period across southern Ontario ranged from 55 to 78 during control periods and from 65 to 84 during heat waves, surpassing levels where mortality is known to increase substantially. Heat stress followed a consistent geographic pattern with the most affected areas in the southern region of the study area, surrounding major metropolitan areas. These HSI maps indicate areas less optimal for dairy farming within the study boundary. Thus some areas currently used for dairy farming and at high-risk for heat stress mortality may require heat abatement strategies to sustain dairy cow production as heat waves become ever more frequent and intense.


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Poster 15 The Role of PRKAR1a in Canine Osteosarcoma Jacqueline Gall ienne, Geoffrey Wood, Jonathon Liu Department of Pathobiology Joint cartilage has poor intrinsic repair potential Canine osteosarcoma (OSA) is an aggressive neoplasm of malignant osteoblasts, and it represents the majority of all primary bone malignancies in the dog. These tumors typically metastasize early in the course of disease and metastatic lesions predominantly develop in the lung. The presence of gross, radiographically detectable metastases confer an especially grave prognosis in canine OSA because chemotherapy tends to be less effective at improving survival at this stage of cancer progression. Some OSA tumours express low levels of the tumour suppressor Prkar1a, commonly known as the R1alpha regulatory subunit of cyclic AMP-dependent protein kinase A. In both human and canine OSA, low Prkar1a-expressing tumours are associated with longer post-chemotherapy survival times compared to tumours with high Prkar1a expression. We hypothesize that low Prkar1a-expressing canine OSA cells will have an inherent greater sensitivity to chemotherapeutic drugs compared to high Prkar1a-expressing tumours. Here, we show three canine OSA cell lines with varying Prkar1a expression levels; the highest level of Prkar1a expression was identified by western blot from primary cells derived from an appendicular case of canine OSA. This cell line was chosen for siRNA-mediated knockdown of Prkar1a in vitro, with the intent to compare chemotherapy responses from the Prkar1a-knockdown treatment to the same cell line with a higher expression of Prkar1a. Ultimately, Prkar1a may provide a potential therapeutic target to sensitize canine OSA tumours to chemotherapy, leading to enhanced treatment efficacy and improved patient survival

Poster 16 Development of a Transdermal Delivery System for a Topical NSAID, Meloxicam Miyuki Kumagai, Ron Johnson, Joe Gabriele, Tom Gibson Department of Biomedical Sciences The purpose of this project is to evaluate a transdermal delivery system, Delivra®, for potential use in veterinary medicine. It is hypothesized that Delivra® will transport meloxicam locally and have equal or greater anti-inflammatory and analgesic effects compared to oral or injectable forms. To determine the efficacy of meloxicam transportation, 3 male and 3 female canines, at 20-40kg and 6-24 mo of age were enrolled in a 2-phase randomized crossover study with oral (0.2mg/kg loading dose, 0.1mg/kg dose q24h for 5 days) or transdermal (1mg/kg dose q24h for 6 days) treatment groups. Plasma, urine, synovial fluid and skin biopsy samples were collected at baseline and 8 hours post last dosing in each phase. To determine analgesic and anti-inflammatory effects of transdermally administered meloxicam, a carrageenan-induced hind-paw inflammation will be induced in male Sprague-Dawley rats, 200-225g. Rats are treated with injectable meloxicam (1.0mg/kg), transdermal meloxicam (1.0mg/kg), transdermal diclofenac (1.0mg/kg), transdermal ASU (1.0mg/kg), Delivra® base or inert cream and paw tissue and serum will be collected for histopathology, inflammatory biomarker gene expression, and drug level analyses. Results from the canine study indicate no adverse systemic effects with oral or transdermal treatment. Additionally, there were no significant differences in systemic (p=0.6290) or local (p=0.7502) meloxicam levels with oral or transdermal treatment. Thus, it can be concluded that transdermal delivery of meloxicam results in equal systemic and local levels when compared to the oral delivery route in the canine stifle. Results from the rat inflammation study are currently being obtained.


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Poster 17 Programmed death l igand 2 gene regulation in the context of lentiviral infection

Beth MacMillan, Olivier Cote, Mary Ellen Clark, Dorothee Bienzle


Programmed death ligand 2 (PD-L2) is a T-cell receptor (TCR) co-stimulatory molecule predominantly expressed on antigen-presenting cells (APCs). PD-L2 expression appears to be highly specific but regulation of its expression has not been well characterized. PD-L2 on APCs interacst with the membrane-bound receptor PD-1 on activated T-cells. This interaction transmits a negative signal to T-cells, modulating their effector function via anergy or apoptosis. Infection with viruses such as the feline immunodeficiency virus (FIV) leads to overexpression of PD-L2, inhibiting appropriate T-cell proliferation and cytokine production, enabling viral persistence. The mode by which FIV influences gene expression of PD-L2 is unknown. Therefore, the hypothesis that FIV interacts with regulatory sequences of PD-L2 to up-regulate expression will be addressed. To test this hypothesis, the upstream region of PD-L2 in the feline genome was amplified and sequenced to identify the promoter region and transcription factor binding sites. Transcription factors that putatively bind and may be involved in regulating PD-L2 gene expression include: STAT6, NF-kB, Oct-1, NFAT and GATA-3. Furthermore, FIV accessory proteins will be interrogated for protein motifs that may bind to the promoter of PD-L2. Promoter constructs will be created to determine the effect of transcription factor and FIV protein binding on expression of PD-L2 using reporter assays. Feline cell lines including FeT-J, MYA-1, Fcwf-4, and FL4 cells, have been examined for suitability in reporter assays. Characterizing regulatory mechanisms of PD-L2 will contribute to understanding the role of this pathway in persistent infection with viruses such as FIV, which in turn may allow design of specific therapeutics to reduce expression

Poster 18 Common confounders of dietary el imination trials contain the antigens soy, pork, and beef Jacqueline M. Parr and Rebecca L. Remillard Department of Clinical Studies Nutritionists and dermatologists recommend avoiding flavored over the counter (OTC) products and medications during dietary elimination trials because these products are thought to contain common proteins that may confound the trial. The objective of this study was to determine if there are soy, pork, and beef antigens in flavored OTC products and medications and if so could these antigens be identified. Seven products, three OTC products and four veterinary therapeutics, were tested using enzyme-linked immunosorbent assays (ELISA) for the presence of soy, pork, and beef antigens, in addition to positive and negative controls. All OTC test products produced ELISA results in agreement with their ingredient lists. ELISA testing of veterinary therapeutic products did not agree with their ingredient lists or product inserts because of other ingredients not listed. Veterinarians should contact manufacturers of oral therapeutics prior to prescribing them to determine other ingredients. Likewise, manufacturers should be contacted regarding 'natural and artificial flavors'. Lastly, gelatin capsules may contain either beef or pork proteins and should not be administered during a trial. In conclusion, flavored medications contain the common antigens soy, pork, and beef although they may or may not be listed on the ingredient list or product insert.


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Poster 19 Pericyte coverage is synchronized with vessel outgrowth during tai l regeneration in the leopard gecko (Eublepharis macularius) Hanna M. Peacock, Jim Petrik, Matt Vickaryous Department of Biomedical Sciences Formation of new tissue requires the simultaneous development of blood vessels to supply the tissue. The most common form of blood vessel formation, known as angiogenesis, begins when an endothelial sprout forms off of an existing vessel and grows into an endothelial tube. Pericytes are then recruited to cover the endothelial tube. Endothelial tubes lacking pericyte coverage may be pruned in response to the endogenous anti-angiogenic Thrombospondin-1 (TSP-1). Pericyte coverage of endothelial tubes is delayed or absent during pathological angiogenesis, (e.g., during tumour formation). As a result, these endothelial tubes are vulnerable to TSP-1. Angiogenesis also plays a role during regeneration, but little is known about the synchrony between endothelial tube formation and pericyte coverage during regeneration. To investigate the relationship between endothelial tube formation and the onset of pericyte coverage, we used an emerging model of regeneration, the leopard gecko (Eublepharis macularius). We hypothesize that during regeneration-mediated angiogenesis, pericyte coverage is synchronized with endothelial tube formation. To determine if endothelial tubes lacking pericytes were present during regeneration, we treated regenerating geckos with a mimetic of TSP-1. This treatment did not impair tail outgrowth or vascularization. To visualize endothelial cells and quiescent pericytes we used immunofluorescence. Quiescent pericytes initially localize to endothelial tubes in the proximo-central regions of the regenerate tail. As regeneration continues, quiescent pericyte coverage spreads to more superficial and distal tail regions. These results indicate that formation of endothelial tubes, and subsequent pericyte coverage, are synchronized during regeneration-mediated angiogenesis.

Poster 20 Targeting Acute Lymphoblastic Leukemia with Oncolytic Virotherapy Christian Ternamian and Byram Bridle Department of Pathobiology B-cell acute lymphoblastic leukemia (B-ALL) is a hematological disease characterized by the rapid expansion and metastasis of malignant lymphoblasts from the bone marrow. Treatment, consisting of chemotherapy, radiation therapy, as well as hematopoietic stem-cell transplantation for high-risk B-ALL patients, is relatively efficacious in children, but cure rates in adults remain relatively low. In both patient populations, central nervous system (CNS) involvement remains an impediment to durable remission. Furthermore, ALL treatment increases the risk of developing adverse health effects, including secondary neoplasms, organ toxicities, and reduced cognitive function. Oncolytic virotherapy is an emerging treatment modality that uses oncolytic viruses (OVs) to selectively destroy tumor cells, while sparing healthy cells. This discrepancy is thought to exist due to defects in the innate cellular immune response that arise during tumorigenesis. As expected of a heterogeneous population, certain tumor cells retain a quasi-functional antiviral immune response that can impede viral oncolysis. Interestingly, treatment with a novel class of epigenetic drugs known as histone deacetylase inhibitors (HDIs) can restore sensitivity to OVs in resistant tumor cells. It is hypothesized that OVs and HDIs will synergistically destroy malignant lymphoblastic leukemia cells. It is also hypothesized that successful induction of immunogenic cell death in tumor cells will elicit an enhanced antitumor immune response. Interestingly, IL-8 has recently been shown to play an important role in promoting immunogenic cell death. My objective will be to investigate the antitumor effects of OVs and HDIs, as well as their ability to induce immunogenic cell death in combination with IL-8.


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Poster 21 Oncolytic immunotherapy for the treatment of high-grade glioma Zafir Syed and Byram Bridle Department of Pathobiology Primary brain cancer, specifically, high-grade glioma (HGG), in human patients has a median survival of 15 months. Despite aggressive treatment, virtually all tumours recur and are fatal. Novel therapies are required. Immunotherapy offers the prospect of killing malignant cells with exquisite specificity. Virus-vectored cancer vaccines represent one strategy to train the immune system to attack tumours. Recent pre-clinical research has shown promise in using vaccines to mount efficacious immune responses against tumours by targeting tumour-associated antigens (TAA). Oncolytic virotherapy involves using attenuated viruses that replicate in and kill cancerous but not normal cells. Recent research in a murine model of melanoma in the brain demonstrated that immunotherapy and oncolytic virotherapy (using Recombinant Vesicular Stomatitis Virus (rVSV)) could be synergized if rVSV expressing a TAA was used to boost rHuAd5-primed tumour-specific T cells. The objective of the present study is to assess the potential of using a rare serotype of adenovirus (i.e. human serotype 48) in combination with MG1 in a homologous prime-boost strategy to treat primary brain cancer. GL261 glioma cells will be implanted into the brains of C57BL/6 mice. Telomerase reverse transcriptase (TERT) is a leading candidate as a TAA. It is hypothesized that treatment of mice bearing GL261 tumours in the brain with rHuAd48-TERT followed by boosting with rMG1-TERT will result in the induction of high-magnitude TERT-specific immune responses that will extend survival.

Poster 22 Canine mast cell tumour (MCT) cells: A model for studying human cancers driven by dysregulation of KIT and VEGFR2 Sean Masson and Brenda Coomber Department of Biomedical Sciences Canine mast cell tumours (MCTs) are a common skin malignancy in dogs, largely driven by mutations in the c-kit gene. Tumours show KIT dysregulation, and the cancer cells signal through PDGFRs and VEGFRs. High grade/metastatic MCTs are resistant to conventional chemotherapeutics, thus canine MCT cells represent a valuable model for KIT dysregulation in human cancers. Better understanding of aberrant KIT (and other RTK) signalling in canine MCT cells could therefore improve design and use of targeted therapies for human cancers driven by c-kit mutations. Canine MCTs are currently treated with the TKIs toceranib or masitinib, with mixed results. Itraconazole, an anti-fungal agent shown to interfere with VEGFR2 glycosylation and hence signalling, may also affect KIT in these cells. The inhibitory effect of itraconazole on two MCT cell lines (MCT1: KIT wt; MCT2: KIT mutant) was evaluated individually and in combination with toceranib. Cell proliferation was quantified in response to itraconazole treatment (0, 100, 200, 400, 800 nM). Increases in proliferation were seen at low doses (100 and 200 nM) with no change at higher doses. Combination treatment with 0.1µM of toceranib did not affect cell growth. However, western blots showed reduced levels of phosphorylated KIT and VEGFR2 in both cell lines. More importantly, a shift towards lower molecular weight native proteins was also seen in MCT-2 cells, indicating itraconazole may interfere with receptor glycosylation in these cells. Itraconazole should therefore be considered a prospective novel therapeutic, warranting further investigation as a treatment for cancers driven by KIT and VEGFR2 dysregulation.


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Poster 23 The Effects of Low-Dose Metronomic Chemotherapy Administration on Ovarian Function and Female Ferti l i ty Jacqueline Dynes and Jim Petrik Department of Biomedical Sciences Chemotherapy preferentially targets rapidly dividing tumor cells, but can also be toxic to normally proliferative cells such as those found in the ovary. Destruction of ovarian cells can lead to early menopause or infertility. Traditional Maximum-Tolerated Dose (MTD) chemotherapy involves administration of high-dose cytotoxic drugs followed by a drug-free interval to allow recovery from harmful side effects. Low-Dose Metronomic (LDM) chemotherapy is an alternative approach where lower doses of chemotherapy are administered more frequently without requiring a period of rest. We hypothesize that LDM chemotherapy will be less toxic to ovarian cells and will not be as detrimental to fertility compared to MTD chemotherapy. Mice were treated with cyclophosphamide using either LDM or MTD chemotherapy scheduling, their ovarian cycles were synchronized, and ovaries were collected at various phases of the ovarian cycle. The ovaries were assessed for morphometric changes and markers of cell proliferation and apoptosis. There were significantly less apoptotic cells in preantral follicles and, based on preliminary follicle counts, the rate of new follicle recruitment also appeared to be lower in the LDM treatment group compared to the MTD chemotherapy group, suggesting less damage to the growing follicle population. These findings were supported in vitro as the viability of cultured rat granulosa cells treated with chemotherapy was significantly higher when treated with more frequent low doses compared to less frequent high doses of chemotherapy. Our results suggest a novel benefit of LDM chemotherapy scheduling that could be implicated in fertility preservation for young females undergoing chemotherapy

Poster 24 Exploring the Impact of Pet-Loss on Pet Owners and Implications for Veterinarian-Client Communication Liam Remillard, Michael Meehan, Jason Coe, David Kelton Department of Population Medicine Research has shown that grief due to pet loss (i.e. sudden death or euthanasia of a pet) is similar to the grief symptoms of people who have experienced the death of a significant other. Support groups and helplines for people who are experiencing grief are commonplace in society. However, only recently have hotlines emerged as a means of providing assistance for pet owners in dealing with the death of their pets. At present there is minimal research evaluating the value of these services and callers’ grief. The present study aims to identify factors associated with grief among the callers to the Ontario Veterinary College Guelph Pet Loss Support Hotline (PLSH). The first stage of research involved content analysis of written notes about reasons why callers contacted the hotline. General themes emerging from the analysis include a lack of social support, strong human-animal bond, symptoms of complicated grief and anger/resentment towards veterinary staff. Furthermore, callers noted behavioural, psychological and somatic challenges resulting from their loss. The results of this study informed the second stage of research involving developing a survey to be sent to hotline callers and members of the general public that have experienced pet loss within the past five years. The survey includes validated measures of grief, pet attachment, social support, general health, and questions about communication with veterinary staff about pet loss. Stage two findings will inform training of PLSH volunteers, improve our understanding of pet loss grief, and develop best-practice guidelines for veterinarian-client communications about pet loss.


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Poster 25 Metabolic modulators as chemotherapeutic sensitizers for colorectal cancer cells

Nelson Ho, Stacey Butler, Brenda Coomber

Department of Biomedical Sciences

Anti-cancer therapy targeting cancer metabolism is being actively explored following its recognition as an emerging hallmark of cancer. Compounds such as dichloroacetate (DCA), arsenic trioxide (ATO), metformin (MET), and 3-bromopyruvate (3BP) are able to alter cellular metabolism and inhibit proliferation of different cancer cell types. Current chemotherapeutic strategies against colorectal cancer generally involve the use of cytotoxic agent 5-fluorouracil (5-FU) in combination with various adjuvants. We therefore investigated the potential synergistic effects of combining 5-FU with different metabolic modulators against human colorectal cancer (CRC) cells. Several human CRC cell lines were treated with 5-FU alone or in combination with different concentrations of DCA (< 10 mM), ATO (< 10 µM), MET (< 10 mM) and 3BP (< 50 µM) for 24 – 72 hr, followed by assessment of cell numbers. We found that cancer cells were protected from the effects of 5-FU when treated in conjunction with the lowest concentrations of each compound tested. This cytoprotection was lost with increasing concentrations of each compound and additionally showed an additive effect in combination with 5-FU. We subsequently assessed apoptosis through caspase 3-cleavage detection to further demonstrate the synergistic potential in combining 5-FU with different metabolic modulators. It is the goal of this project to unravel therapeutic avenues that would lead to lower chemotherapy doses by combining them with metabolic adjuvants to help improve patient outcome while reducing the toxicity and subsequent side effects associated with current chemotherapeutic strategies in the clinical setting.

Poster 26 The effect of growth conditions on the expression of Actinobacil lus suis adhesin genes Adina Bujold and Janet MacInnes Department of Pathobiology Actinobacillus suis (A. suis) is a common commensal of the tonsils of the soft palate of swine, but in the presence of unknown stimuli it can invade the bloodstream and cause systemic disease. Its pathogenesis, including host colonization, is poorly understood. Thus, the objective of this study was to measure the expression of genes involved in attachment to tonsils under various growth conditions. From 40 adhesin genes identified by bioinformatics in a clinical isolate of A. suis (encoding 22 putative adhesins), 12 genes were chosen for analysis by qPCR. Aerobic cultures were grown at 37ºC with shaking at 200 rpm, in the presence and absence of 50 µM epinephrine. RNA was extracted from samples collected at exponential and stationary phases of growth, and qPCR conducted. The presence of physiological levels of epinephrine did not have a significant effect on expression of most adhesin genes tested, while significant differential expression of several adhesin genes was observed in response to growth phase. Type IV fimbrial subunit ppdD, fine tangled pili ftpA, and outer membrane protein ompA were up-regulated in stationary growth vs. exponential growth, while tight adherence (tad) genes tadD and tadG, fibronectin-binding ybaV, ompP2, and filamentous hemagglutinin (fha) transporter fhaC were down-regulated. Interestingly, fimbrial components of the tad locus and the fha were not significantly differentially expressed. To model the environment of tonsillar crypts, work is underway to assess the effect of anoxic static growth on expression of these adhesin genes. This should provide insight into how A. suis and other pathogens invade tonsils.


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Poster 27 Evaluation of a handheld device for the detection of β -hydroxybutyrate pre-calving in dairy cattle

El ise Tatone, Jessica Gordon, Stephen LeBlanc, Todd Duffield

Department of Population Medicine

Individual and herd ketone concentrations are commonly monitored in adult dairy cattle post-partum to identify individuals at-risk for metabolic disease and to identify potential improvements to management factors. The Precision Xtra® handheld meter has been validated for use in dairy cattle for b-hydroxybutyrate (BHBA) measurements post-calving as a convenient cow-side test for ketonemia. Non-esterified fatty acids (NEFAs) can be measured pre-partum to help predict post-partum risks earlier. However, this test is inconvenient and time consuming as samples must be processed and sent to a laboratory for evaluation. Recent research has identified BHBA cut points pre-partum of 0.6-0.8 mmol/L associated with increased risk of post-partum disease. The objective of the current research was to compare results of the Precision Xtra® handheld device with laboratory evaluation of serum BHBA pre-calving. The results of the Precision Xtra® were compared to laboratory BHBA and NEFA concentrations and evaluated by looking at concordance coefficients, sensitivity, specificity and Receiver Operator Characteristic curves at cut points of 0.6-0.9 mmol/L. The two tests had a moderate concordance correlation of 0.77 and the area under the curve for each cut point was high with values between 0.90-0.93. The Precision Xtra® had sensitivities of 84.6-92.8% and specificities of 75.5-98.5% depending on the cut point evaluated. Based on the moderate level of correlation and the good level of sensitivity and specificity, the Precision Xtra® is a useful tool in the detection of elevated BHBA pre-calving and may be helpful in identifying individuals at risk for post-partum disease.

Poster 28 Cleavage and blastocyst rates, and sex ratio are altered by oocyte exposure to bisphenol A in Bos Taurus Jacqueline Ferris, John Leatherland, Laura Favetta, Neil MacLusky, Al lan King Department of Biomedical Sciences Bisphenol A (BPA), an endocrine disrupting chemical, can have detrimental effects on fertility and reproductive success, having been linked to reproductive issues such as reduced in vitro fertilization (IVF) success in humans and recurrent miscarriages. Feminization of males and a sex skew towards females have been found in fish and amphibians in vivo as a result of early exposure to BPA. The current study evaluated the effects of BPA exposure during in vitro bovine oocyte maturation on embryo cleavage and blastocyst development rates as well as the sex ratio of resulting embryos surviving to blastocyst. To test our hypothesis that cleavage and blastocyst rates would be diminished, and sex ratio would be skewed towards females, oocytes were matured in in vitro maturation (IVM) media under various treatments (BPA concentrations of 3 and 30 µg/mL) and controls (no-treatment, vehicle and estradiol). Standard bovine IVF procedure was completed following maturation. Cleavage rates in the 30 µg/mL BPA treatment group were significantly lower compared to the no-treatment and vehicle controls. The proportion of cultured embryos to reach the blastocyst stage were significantly lower in the 30 µg/mL BPA treatment group compared to the no-treatment and vehicle controls. Oocyte treatment with 30 µg/mL BPA resulted in a higher number of females surviving to blastocyst compared to the no-treatment control. These results suggest that bovine oocyte exposure to BPA can compromise the resulting embryo's early developmental competence, and influence sex ratio of surviving blastocysts, thereby detrimentally impacting bovine reproduction.


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Poster 29 Telomere and Telomerase Analysis in Bovine Preimplantation Embryos Graham Gilchrist, W. Al lan King, Jon LaMarre Department of Biomedical Sciences Telomeres are tandem repeats of nucleic acid and specialized proteins that stabilize the ends of chromosomes from erosion and fusion with self or other chromosomes. Telomeres shorten with each round of cellular division and a build up of critically short telomeres induces a state of replicative senescence and therefore are considered a biomarker for aging and age-related pathologies. It has been hypothesized that the rate of telomere attrition differs in sexes and alternately, the differences in age-related pathologies is attributed to this difference in telomere length. However, little is known of any sex specific difference during in telomeres in embryo development. The purpose of this study was to identify the changes in telomere length and telomerase activity in developing embryos, including the differences between male and female bovine in vitro produced embryos. Embryos were produced using standard in vitro production procedures and the oocytes and embryos were collected at various time points from the germinal vesicle oocytes (GV), to the day 8 blastocysts stage. Relative telomere length was determined using real-time quantitative PCR (qPCR), and telomerase activity was determined using a qPCR-based assay. There is no statistically significant difference between male and female embryo relative telomere lengths at the various stages of development, however, there was a U-shaped trend seen across all stages. Relative telomerase activity was low from the oocytes to the 8-cell stage, and increased significantly at the blastocyst stage on a per embryo basis. These results indicate that telomere lengths are reset during embryogenesis and any differences in telomere attrition seen between males and females is not occurring during embryo development and occurs post-natal.

Poster 30 Effect of Hoof Orientation and Ballast on Acceleration and Vibration in the Hoof and Distal Forelimb Following Simulated Impacts Ex Vivo . Cristin McCarty, Jeffrey Thomason, Karen Gordon, Timothy Burkhart Department of Biomedical Sciences Reasoning: To establish baseline data for impact loading of the distal limb as a precursor to assessing the potential role of impact in injury and joint disease. Objectives: Examine the effect of three hoof-strike conditions (toe-first, flat, heel-first) and two specimen masses (with and without a ballast of approx. 2% body mass) on impact deceleration and vibration frequencies and energies at the hoof, first phalanx (P1) and third metacarpal (MC3). Design: Eight cadaver limbs were subjected to randomized, repeated controlled trials in which the hoof was struck by a pendulum impact testing machine (impact velocity 3.55m/s) under the three strike and two mass conditions. Methods: Data from three-axis accelerometers on the hoof, P1 and MC3 quantified, for all trials, the peak impact acceleration, frequencies in the first 6.4ms following impact, the frequency with the most energy, 95% of the total energy, and the frequency at 95% cumulative energy. The effects of the strike and mass conditions on each variable were statistically tested using repeated-measures ANOVA (α=0.05). Results: Signal energy reaching MC3 was 6-31% of that at the hoof. A heel-first strike produced the largest peak accelerations and highest frequencies among all strike conditions, and changing the mass had no effect regardless of strike condition. Conclusions: Large accelerations that occur upon impact of the hoof with the ground are attenuated by the distal structures of the equine limb, but still carry considerable energy within the signal that could be damaging to tissue and are dependent on hoof strike condition but not ballast. Potential relevance: Our results suggest that impact loading on the hoof could be a factor in contributing to bone injury and joint disease in the distal limb.


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Poster 31 High IL-1beta expression in peritumoural stromal cells is associated with more aggressive tumours in an inflammatory mouse model of prostate cancer Christopher Pinell i , Hibret Adissu, Ir is Fang, Rama Khokha, Geoffrey Wood Department of Pathobiology Chronic inflammation is a contributing factor in the development of a number of different cancers, including prostate cancer. Tissue inhibitor of metalloproteinase-3 (TIMP3) is an endogenous inhibitor of inflammation and loss of TIMP3 has been documented in human prostate cancer, although the role of this gene in prostate cancer progression is not established. Since TIMP3-deficient mice show uncontrolled/enhanced inflammation in a variety of disease models, we crossed prostate-specific Pten homozygous knockout mice (ARR2Pb-Cre Pten flox/flox), an established mouse model of prostate cancer, into a Timp3-/- background to investigate the role of inflammation in prostate cancer development and progression. Timp3-/- mice had shorter tumour latency with increased peritumoural inflammatory cells, and increased tumour mRNA expression of COX-2, IL-1beta and monocyte chemoattractant protein-1 (MCP-1) compared to wildtype (WT) mice. Multiplex bead-based ELISA analysis of tumour homogenates revealed significantly higher IL-1beta and MCP-1 protein expression in Timp3-/- compared to WT mice. Immunohistochemistry for COX-2, IL-1beta and MCP-1 demonstrated negative labelling of tumour cells for all three inflammatory mediators, but positive cytoplasmic labelling of IL-1beta in peritumoural bone marrow-derived (BMD) cells and tumour-associated fibroblasts in all mice. There were significantly more BMD cells labelling positively for IL-1beta in Timp3-/- compared to WT mice (2.20±0.28 versus 1.10±0.25 % positive cells per 600x field respectively, p<0.05), and there was significant correlation between IL-1beta IHC scoring and multiplex ELISA data across all genotypes (r2=0.737, p<0.05). Thus a pro-inflammatory microenvironment, specifically BMD cells expressing IL-1beta, is associated with enhanced progression to invasive prostate cancer in this mouse model.

Poster 32 Understanding metabolomic differences between bovine embryos of different developmental competence

Kayla Perkel, Pavneesh Madan Department of Biomedical Sciences Introduction: In vitro, the most common and preferred method for evaluating an embryos developmental potential is by ascertaining the cleavage rate coupled with morphological assessment, in spite of the inherent limitations and inaccuracies of these types of evaluations. A recent shift in the literature has aimed to find alternative technologies, such as through the study of metabolomics, to identify embryos with the highest developmental potential through culture media analysis. We hypothesize that embryos developing at different rates differ in their metabolomic signature. The objective of this study was to determine the metabolomic signatures of fast (FG) and slow (SG) growing embryos at timed stages of development. Methodology: Standard IVM and IVF protocols were used on oocytes aspirated from abattoir obtained ovaries. Presumptive zygotes were placed individually in 40 µl culture drops. Media from FG embryos was collected at 2-cell (30 hours post fertilization or hpf), 4-cell (42 hpf), 8-cell (49 hpf), 16-cell (90 hpf), morula (144 hpf) and blastocyst (168 hpf) and from SG embryos 12 hr later to reach equivalent embryo stage. Nuclear magnetic resonance was performed on a 600MHz spectrometer. Results: Data indicates distinct differences in metabolomic signatures between SG and FG embryos using multivariate statistical analysis. Specifically, production and consumption of metabolites like pyruvate, lactate, alanine, glutamate, valine, leucine and isoleucine differed between SG and FG embryos. Conclusion: The results provide a first step towards the use of metabolomics for the development of a non-invasive tool for assessing embryo viability and the discovery of potential biomarkers.


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Poster 33 Exploring Strategies for Johne’s Disease Prevention in Organic Dairy Herds in Ontario, Canada Laura Pieper, Ann Godkin, Ulrike Sorge, Kerry Lissemore, Trevor DeVries, David Kelton Department of Population Medicine Johne’s disease (JD) or paratuberculosis is a chronic, infectious, gastrointestinal disease in cattle caused by Mycobacterium avium spp. paratuberculosis. To minimize the effects of this disease, the Ontario dairy industry implemented a risk assessment based JD control program in 2010. Local veterinarians fill out an on farm management questionnaire and based on that, give recommendations for JD prevention. However, organic farmers might not be able or willing to implement those recommendations due to their regulations or beliefs. Therefore, the objective of this study was to explore strategies for JD prevention on organic dairy farms in Ontario, Canada. Semi-structured interviews (individual interviews: n=3, focus groups: n=3) have been conducted between April and June 2013 with a total of 16 Ontario veterinarians and organic dairy farmers. The interviews were audio-recorded, transcribed and analyzed using thematic analysis. Farmers and veterinarians agreed that there might be hesitation among organic farmers to change the major aspects of their calving and calf management in the recommended way. However, they also mentioned certain risk reducing behaviors that could be implemented more easily, such as increasing barn cleanliness and keeping a closed herd. The interviewees identified risk areas that are not captured by the risk assessment and expressed their perceived lack of knowledge about several topics concerning JD. With this information we would be able to modify the risk assessment to more precisely capture the possibility for disease transmission on organic farms and to provide veterinarians with recommendations for farms that work according to organic management standards.

Poster 34 A cytogenetic study of young breeding boars in Canada Anh Quach, Revay Tamas, Mariana Macedo, W. Allen King Department of Biomedical Sciences The fact that chromosome rearrangements are major etiologic factors behind subfertility has been shown in many studies, which have revealed three major types of abnormalities; reciprocal translocations, centric fusions and inversions. It is also observed that, although not fully explained, “de novo” chromosome rearrangements occur more frequently in pig than in other species. In our previous pilot study (TA Quach et al. 2009) we showed that chromosome abnormalities in the Canadian swine population are very high (2.5%). Therefore, the aim of this study is to detect the potential carriers of chromosomal anomalies in a much larger sample size. In total, 300 young boars from 4 different breeds (Duroc, Landrace, Pietrain and Yorkshire) were karyotyped by G-band techniques. Four previously unreported reciprocal translocation including rcp(1;5), rcp(3;4), rcp(8;13) and rcp(7;15) and one Robertsonian translocation rob(13;17) were found, thus the frequency of chromosome abnormalities in this study is 1.67%. We are able to determine that rcp(3;4) and rob(13;17) were inherited from their dams and rcp(8;13) was a “de novo” event. Prolificacy of rcp(3;4) and rcp(7;15) translocation carriers was noted to be reduced (24% and 38%, respectively) while it was slightly reduced 9% in carriers of rob(13;17). More studies need to be carried out to further investigate the effect of these translocations.


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Poster 35 Antimicrobial resistance in Salmonella and generic E. coli isolated from raccoons and skunks and their environments

Mackenzie Slif ierz, Jeonghwa Park, Robert Friendship, Scott Weese


Staphylococcus hyicus is the causative agent of exudative epidermitis (greasy pig disease) in pigs which is characterized by skin lesions and a greasy exudate. The objective of this study was to determine whether methicillin-resistant S. hyicus (MRSH) carries the zinc-resistance gene (czrC). Twenty-eight MRSH isolates from 28 pigs divided across 15 different farms were screened for czrC using PCR. Fourteen (50%) pigs were colonized with MRSH that carried czrC, representing 8 (53%) of the 15 farms from which MRSH was isolated. The finding that czrC is common amongst MRSH and distributed across multiple Ontario farms raises concerns because pigs are commonly exposed to high levels of in-feed zinc oxide. This exposure may cause selection of these multidrug resistant bacteria carrying czrC, and make treatment more difficult and costly for exudative epidermitis infections. In addition, of the pigs carrying czrC positive MRSH, 64% (9/14) were reported to be raised without exposure to antibiotics, and this association was statistically significant (P<0.05, Fischer’s Exact). This association was foreseeable as zinc oxide is a popular alternative to in-feed antibiotics, and zinc-resistance may explain the persistence of multidrug-resistant staphylococci on antibiotic-free farms. Further research is needed to explore the effect of high levels of in-feed zinc oxide on methicillin-resistant staphylococci in pig production.

Poster 36 Isolation, immunophenotyping and lymphocyte suppressive properties of equine CB-MSC Laurence Tessier, Dorothee Bienzle, Lynn B Wil l iam, Thomas G Koch


Multipotent mesenchymal stromal cells (MSC) have attracted interest for their potential use in cytotherapy, due in part, to their ability to secrete immunosuppressive factors. We hypothesized that MSC can be consistently isolated from equine cord blood (CB), have unique and reproducible marker expression, which includes cytoplasmic Toll-like receptor (TLR) 3 and TLR4, and in vitro suppress lymphoproliferation. Isolation success, progenitor frequency and cell population doubling times were calculated from 9 CB samples. Five additional CB-MSC samples were phenotyped using flow cytometry and quantitative polymerase chain reaction (qPCR) assays. CB-MSC immune regulatory properties were evaluated in mixed lymphocyte reactions (MLR) (n=4). TLR3/4 expression of untreated, poly(I:C)-, or LPS-treated CB-MSC was determined with qPCR and immunocytochemistry (ICC) (n=4). CB-MSC isolation success was 100% (9 of 9 samples). Progenitor frequency ranged from one MSC per 1.61 to 22.5x105 nucleated cells. The cell-doubling times were as follows: passage 2 to 3 (n=9): 1.67 ± 0.15, passage 3 to 4 (n=9): 1.68± 0.15, passage 4 to 5 (n=5): 1.89 ± 0. 58 days. Cultured CB-MSC expressed CD29, CD44 and CD90 but not MHC I, MHC II, CD4, CD8, CD11a/18, and CD73 before and after cryopreservation. CB-MSC significantly suppressed lymphocyte proliferation in MLR. CB-MSC constitutively expressed TLR4 as detected by ICC and qPCR. In conclusion, equine CB is a reliable source of MSC that are cryo-tolerant and express a stable phenotype in culture. Mechanisms of lymphosuppression via TLR3/4 or other molecules warrant further investigation.


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Poster 37 Characterization of PIWI-l ike proteins in early bovine embryogenesis Stewart Russell , Jonathan LaMarre Department of Biomedical Sciences PIWI proteins comprise a subfamily of the Argonaute proteins that bind to specific 26-32 nt RNAs (piRNAs). They are required for the suppression of retrotransposons in the mammalian male germline, with additional uncharacterized roles likely in the gametes of both sexes. Although the majority of research has focused on gonadal functions of PIWI proteins, recent studies have demonstrated the presence of these proteins and their associated piRNAs in early embryos. Bovine embryo development is an economically important process that also represents a translational model for human development. We postulate that PIWI-like proteins are present in bovine early embryos and that they participate in the regulation of gene expression during the reprogramming events of embryogenesis. Four predicted sequences encoding PIWI proteins are present in the bovine genome. We performed RT-PCR with multiple primer sets for each predicted gene using testes, ovaries and oocytes as the tissue sources. Transcripts for PIWI-like proteins 1 and 2 were identified in the testis and ovary, as well as immature and mature oocytes. PIWIL1 protein was detected using antibodies in bovine testis, ovary, and early embryo stages. Finally, two different transcript variants of bovine PIWIL1 were successfully cloned from a testis cDNA library. These data demonstrate that a subset of the PIWI-like proteins are present in early bovine embryos and suggest important roles such as the maintenance of genome integrity during this critical period of development. Future studies will investigate the piRNAs bound to PIWIL1 and PIWIL2, the expression profile of potential targets, and the roles these molecules play in embryo development.

Poster 38 Anti -chemotactic effects of canine protein C are dependent on the endothelial protein C receptor (ECPR) expressed in canine neutrophils Valerie M Wong, Oliver Côté, Dorothee Bienzle, M. Anthony Hayes, R. Darren Wood Department of Pathobiology We previously purified and characterized canine protein C zymogen (CnPC) but little was known about canine EPCR. The goals of this study were to determine if EPCR was expressed in canine neutrophils and to determine the role of EPCR in neutrophil chemotaxis. EPCR cDNA was amplified by PCR from RNA of neutrophils. By flow cytometry, a slight but significant increase in mean fluorescence (p=0.003) was detected in neutrophils incubated with RCR-379, an anti-human protein C antibody, compared to controls. Neutrophils were labelled with calcein AM and incubated with either CnPC or canine activated protein C (CnAPC) (0.1, 1, 10, or 100 ng/mL), with or without RCR-379 (5 mg/L). Treated neutrophils were then allowed to migrate through a filter membrane towards interleukin-8 or complement component C5a. Untreated neutrophils served as positive control. Migration was quantified by fluoresence measurement and expressed as chemotaxis index (fluorescence of test sample/fluorescence of positive control). Without RCR-379, chemotaxis indices for cells treated with either CnPC or CnAPC at all tested concentrations were all significantly <1 (p=0.0002 to 0.007). With RCR-379, chemotaxis indices for cells treated with either CnPC or CnAPC at all but the lowest concentration tested (0.1 ng/µL) were not significantly different from 1 (p=0.08 to 0.7). The effects of RCR-379 on migration were independent of activation status of protein C. In summary, we showed that (1) EPCR was expressed in canine neutrophils and that (2) the anti-chemotactic effects of CnPC or CnAPC were dependent on EPCR and distinct from anticoagulant properties.


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Poster 39 Platelet lysate as alternative to fetal bovine serum in equine mesenchymal stromal cell culture Keith Russell and Thomas Koch Department of Biomedical Sciences Expansion media for mesenchymal stromal cells (MSCs) is typically supplemented with fetal bovine serum (FBS) to provide growth factors necessary for the cells to thrive. However, potential problems associated with FBS are high content variability, increasing costs, and risk of immunorejection of transplanted cells from introduced xenogeneic substances. An alternative to FBS showing potential in human studies is platelet lysate (PL), a concentrated solution of growth factors released from platelet-rich plasma (PRP) by a freeze/thaw procedure. The hypothesis of the study was that equine MSCs expanded in PL-supplemented culture would proliferate at equal rates to those in FBS-supplemented culture while maintaining MSC morphology. The objectives were to generate PRP and PL, determine how equine MSCs grow in medium enriched with PL at various concentrations, and compare PL to FBS. We were successful in concentrating platelets by 4-5 times. A single freeze/thaw cycle released platelet-derived growth factor (PDGF-BB) from platelets to levels 20 times that of PRP. For MSC culture, PL appears to outperform FBS at concentrations up to 20%. Beyond this threshold, PL appears to become detrimental to MSCs, while FBS continues to follow a dose-dependent response with increased cell proliferation at higher FBS concentrations. These results suggest PL is a feasible alternative to FBS in MSC expansion culture. The impact of PL on MSC differentiation potential and other physiological functions warrants investigation. From a clinical perspective, the suggestion of a PL safety margin raises questions about an increasingly common tissue regenerative therapy: the concurrent use of PRP and MSCs.

Poster 40 Variation in miR-34 family expression in bovine gametes and early embryos Al l ison Tscherner, Natasha Smith, Jon LaMarre Department of Biomedical Sciences Differentiated gametes are haploid cells that fuse to form zygotes in sexually reproducing organisms. During gamete maturation transcription is silenced, providing a unique environment for the study of RNA biology in which post-transcriptional regulatory mechanisms predominate. The microRNA-34 family (miR-34 a, b, and c) is found in gametes of several species and appears important for gametogenesis and embryogenesis. In somatic cells, miR-34 is a component of the p53 network, where it impedes proliferation by silencing oncogenic targets. Its role in reproduction is incompletely understood and may be species-specific. In mice miR-34c is abundant in sperm, absent in oocytes and required for the first zygotic cleavage event. In contrast, miR-34 is present in oocytes of Drosophila and Danio. We have investigated the miR-34 family in bovine gametes and early embryos using qRT-PCR to examine miRNAs, their precursors and a known mRNA target. miR-34a, b, and c were detected in spermatozoa, with miR-34c demonstrating the highest relative expression although levels varied greatly between isolates of individual bulls. miR-34a and c, but not b, were present in oocytes and 2-cell embryos. Interestingly, the primary transcript of the miR-34b/c bicistron is abundant in testes (which contain developing spermatocytes) but was undetectable in mature spermatozoa and oocytes, suggesting that processing occurs before completion of meiotic maturation. Finally, synaptotagmin1 mRNA increased markedly during oocyte maturation, suggesting that the expression of this miR-34 target can escape miRNA-dependent repression in oocytes. The presence of the miR-34 family in bovine gametes implicates it in fertility, and variation in miR-34c levels between bulls offers potential for its use as a biomarker of male reproductive competence.


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Poster 41 Diurnal gene expression differences as biomarkers of heart disease in a pressure overload-induced cardiac hypertrophy murine model. Elena Tsimakouridze, Marty Straume, Peter Podobed, Heather Chin, Jon LaMarre, Ron Johnson, Monica Antenos, Gordon Kirby, Al l ison Mackay, Patsy Huether, Jeremy Simpson, Michael Sole, Gerard Gadal, Tami Martino Department of Biomedical Sciences The integrity of circadian gene expression underlies cardiovascular health and disease, however time-of-day profiling of genomic biomarkers in cardiovascular disease (CVD) has never been examined. We use a time-of-day chronomics approach to investigate global gene expression changes in CVD using microarrays and bioinformatics on samples collected over 24hrs. We used a Transverse Aortic Constriction (TAC) murine model of pressure-overload cardiac hypertrophy. After 1 (acute) and 8-wks (chronic), SHAM vs TAC hearts were collected every 4h for 24hrs.RNA was isolated for microarrays and RTPCR. We created a de-novo algorithm, DeltaGene, to identify circadian genes from microarray data as potential biomarkers of CVD. A second set was used to purify proteins for Western blot. Results: The top 300 DeltaGene biomarker candidates were pared to 20 genes, using knowledge-based platforms, for RTPCR validation. The significant time-of-day changes were: uncoupling protein 3 (Ucp3), pyruvate dehydrogenase kinase 4 (Pdk4), E1B interacting protein (Bnip3), solute carrier family25 member22 (Slc25a22), and lipase hormone sensitive (Lipe). We tested whether time-of-day biomarkers could be indicative of disease progression by comparing 1- vs. 8-week TAC, and found unique profiles for Pdk4, Lipe, Ucp3, Slc25a22, Bnip3, thymosin beta10, kinesin5B, and tissue inhibitor of metalloproteinase 2. Because protein expression is functionally relevant, we identified time-of-day profiles for analogous cardiac proteins TIMP2, PDK4, and LIPE. Rhythmic gene and protein expression may have clinical applications for biomarker discovery. This approach is generally applicable to disease and can lead to new discoveries to benefit patients.

Poster 42 Ependymal cells express neural stem/progenitor cell markers during spinal cord regeneration in the leopard gecko (Eublepharis macularius) Emily Gilbert and Matt Vickaryous Department of Biomedical Sciences

The leopard gecko (Eublepharis macularius) is an innovative model for the study of spinal cord regeneration. Unlike mammals, the leopard gecko is able to fully regenerate its spinal cord following amputation. Although the source of endogenous neural stem/progenitor cells (NSPCs) giving rise to the new spinal cord remains unclear, ependymal cells are commonly implicated. Ependymal cells are specialized neuroglia lining the central canal of the spinal cord. To investigate the role of ependymal cells during spinal cord regeneration in the leopard gecko, we conducted a spatiotemporal characterization of several markers of NSPCs, including Sox2, Sox9 and Vimentin using immunohistochemistry/immunofluorescence. In addition, we used PCNA to identify proliferating cells. Our data reveals that prior to injury, ependymal cells are immunonegative for all NSPC markers with only a small population demonstrating evidence of proliferation. Immediately following tail loss, the majority of ependymal cells are PCNA+, and a subset of ependymal cells begin to express Sox2 and Vimentin. Sox2 and Vimentin expression increases throughout regeneration. Sox9 expression does not begin until the later stages of regeneration, and is restricted to a limited subset of ependymal cells. Our findings point towards ependymal cells as a key source of NSPCs during spinal cord regeneration.


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Poster 43 The Potential for Raccoons to Mechanically Transmit Salmonella Kristin Bondo, Samantha Kagan, Nicol Janecko, David Pearl, Richard Reid-Smith, Patrick Boerl in, Jane Parmley, Claire Jardine Department of Pathobiology The potential role of raccoons in transmitting Salmonella biologically (through feces) and mechanically (on paws) is unknown. We isolated Salmonella from rectal and paw swabs of raccoons and from soil and livestock manure lagoon samples in areas where raccoons were trapped on swine farms and conservation areas in southern Ontario from June – October 2012. Salmonella was isolated from 26% (117/476) of raccoon rectal swabs, 18% (56/418) of raccoon paw swabs, 9% (36/497) of soil samples, and 35% (4/13) of livestock lagoon samples. The prevalence of Salmonella was significantly greater in raccoon rectal and paw swabs than in soil samples (p < 0.001 and p = 0.006, respectively); however, the prevalence of Salmonella from rectal swabs was significantly greater than paw swabs (p < 0.001). Of 418 raccoons, 40 (9.6%) tested positive for Salmonella from both paw and rectal swabs and 35 (8.4%) tested negative from rectal swabs but positive from paw swabs. Of 21 Salmonella serovars isolated, 9 were found only in rectal swabs, 4 only on paws, 1 only in manure, and none were unique to soil. Overall, serovars Oranienburg (68/236), Typhimurium (43/236), and Newport (38/236) were the most commonly detected and resistance was found in 4/236 Salmonella isolates. Salmonella serovars in raccoon rectal swabs did not always match what was found on paws from the same individual or from samples in their environment. Our results suggest that raccoons have the potential to transmit Salmonella biologically and mechanically, although the source of Salmonella exposure for raccoons was not clear

Poster 44 Tissue tropism and transduction efficiency of adeno-associated virus (AAV) vectors pseudotyped with AAV capsids isolated from domestic animal sources Darrick Yu, Kelsie Jagt, Jenny Stodola, Kevin Stinson Department of Pathobiology Adeno-associated virus (AAV) is a small, single stranded DNA virus commonly associated with adenovirus infection. AAV is not associated with any known malignancy and thus vectors derived from AAV make promising gene transfer vectors. AAVs derived from animal species may be especially attractive due to a lack of pre-existing immunity within the human population. Animal tissues and animal derived adenovirus stocks were PCR screened for the presence of AAV sequences. Two such sequences were isolated and analyzed: one a bovine AAV related sequence derived from a caprine adenovirus stock, and the other a porcine AAV sequence derived from pig colon. The capsid sequences of each were characterized with respect to amino acid differences from previously reported capsid sequences. The bovine capsid sequence isolated from a caprine adenovirus stock possessed 2 amino acid changes from the previously described bovine AAV capsid sequence, and the porcine capsid possessed 13 amino acid changes as well as a novel four amino acid insertion not previously described. These two capsids may have novel transduction properties due to their divergence from previously reported capsids.


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Poster 45 Testis specif ic protein Y encoded (TSPY) copy number in various bull t issues Olutobi Oluwole, Anh Quach, Tamas Ravey, Laura Favetta and Allan King Department of Biomedical Sciences Testis specific protein Y encoded (TSPY) is a multi-copy gene present on the male specific region of the Y chromosome. It is involved in the normal physiological function of the testes as a proliferation factor during spermatogenesis and thus has been implicated in spermatogenesis and fertility. It is expressed in gonocytes and spermatogonia of embryonic testes as well as in spermatogonia and spermatocytes of adult testes and in some types of tumors. Studies have shown that bulls have from 50-200 copies of TSPY. The objective of this study was to determine whether TSPY copy number varies among different tissues within individual bulls. Blood and other tissue samples (such as seminal vesicles, liver and muscle) were collected from 6 mature bulls. DNA was extracted from the blood and tissues using the phenol chloroform method. The DNA was analyzed by quantitative real time PCR to determine TSPY copy number. Preliminary results show that TSPY copy number varies among the different tissues analyzed compared with whole blood. This study has shown that not only does TSPY copy number vary among individual bulls but also among different tissues within individuals. Further investigations are needed to confirm and determine the cause of the difference in TSPY copy number in various tissues. This study will provide more information on the role of TSPY copy number variation in male fertility.

Poster 46 Ratio of serum and foll icular f luid anti -Müllerian hormone as a predictor of patient embryo uti l ization and pregnancy Anja Stojsin, Michael Neal, Shilpa Amin, Edward Hughes, Mernoosh Faghih, Megan Karnis, Daniel Gil l is, Al lan King Department of Biomedical Sciences

This study was conducted to evaluate the strength of the serum (SE) and follicular fluid (FF) anti-Müllerian hormone (AMH) ratio as a predictor of a patient embryo utilization probability (EUP), and pregnancy. SE and FF AMH levels were measured (pmol/L) and compared to cycle characteristics and outcomes of women (n=58) undergoing IVF at a university affiliated fertility clinic. Data were controlled for patient age, oocytes retrieved, zygotes created, and embryo quality. EUP was calculated as a measure of embryo competence. Logistic regression method was used to investigate the relationship between the response and predictor variables. Pregnancy outcome was compared by t-test or chi-square analysis where appropriate, with p<0.05 considered statistically significant. SE:FF AMH ratio was inversely correlated with EUP and was statistically significant (p=0.0252; 95% CI: -0.0732, -0.0048). Pregnant (P) and non-pregnant (NP) patients had a similar mean SE AMH levels (13.04 vs 14.33, p=0.332). However, FF AMH levels were higher in patients with a positive pregnancy (26.62) compared to their NP counterparts (11.68; p=0.051). When the AMH measured in FF was higher than the SE AMH (FF:SE ratio≥1.5) the pregnancy per cycle started was 70.0% compared to 47.4% in patients with a FF:SE ratio<1.5 (p=0.1). By knowing the ratio of SE to FF AMH concentrations and the patient age, it is possible to calculate the overall EUP that could be used to optimize the overall number of embryos that need to be created, transferred or frozen. In turn, this may enhance IVF effectiveness by clarifying often difficult embryo transfer decisions.


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Acknowledgements

Graduate students at this event presented research projects supported by: ( in alphabetical order):

Agriculture Adaptation Council

Agriculture and Agri-Food Canada

Angell Animal Medical Center

American Association of Bovine Practitioners

Animal Health Laboratory

Animal Health Strategic Investment

Atlantic Innovation Fund and OVC Pet

Trust Fund

Association of Local Public Health Agencies

Canadian Food Inspection Agency

Canada Research Chairs program

Canadian Bovine Mastitis Research Network

Canadian Cancer Society Research Institute

Canadian Centre for Swine Improvement Inc.

Canadian Cooperative Wildlife Health Centre

Canadian Dairy Network (DairyGen Council)

Canadian Institutes of Health Research

Canadian Poultry Research Council

Canadian Swine Health Board

Chicken Farmers of Ontario

Chilean Government

Dairy Farmers of Canada

The Danish Research Agency for Technology, Innovation and Production

Dairy Farmer’s of Ontario Doctoral Research Assistantship

Danish Agency for Technology and

Production and Innovation Equine Guelph

Hamilton Public Health Services

International Livestock Research Institute

Natural Sciences and Engineering Research Council

Nick Natale Innovation Grant of the Canadian Cancer Society

NSERC Collaborative Research and Development

NSERC Discovery Grant

NSERC Idea to Innovation

Ontario Cattleman’s Association

Ontario Graduate Scholarship

Ontario Ministry of Agriculture, Food and Rural Affairs


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Ontario Ministry of Research and Innovation

Ontario Ministry of Health and Long-term

Care

Ontario Sheep Marketing Agency

OVC Fellowship and Scholarship Fund

OVC Incentive Fund

OVC Pet Trust Fund

Poultry Industry Council

Public Health Agency of Canada

Prostate Cancer Canada

The XXIII World Veterinary Congress Foundation and Aeroplan

USDA National Institute of Food and Agriculture Grant

Vanier Canada Graduate Scholarships

Veterinarians without Borders

Vietnamese Overseas Scholarship Program


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Graduate Student Research Symposium Organizing

Committee

Dr. Gordon Kirby, Associate Dean, Research and Innovation Barb Gaudette, Research and Graduate Affairs Assistant

Elizabeth Lowenger, Manager, Student Affairs Andreia Arruda, Population Medicine Robbyn Sargent, Population Medicine

Kelsey Gahnsmith, Biomedical Sciences Courtney Schott, Pathobiology

Shawn MacKenzie, Clinical Studies Faisal Alibhai, Biomedical Sciences

Special Thanks to Faisal Al ibhai (Biomedical Sciences) for preparing the proceedings booklet

Oral & Poster Presentation Judges

Faculty & Post-Doctoral Fellows

Byram Bridle Brenda Coomber

Marie Holowaychuk Bettina Kalisch Brandon Lillie John Lumsden Paula Menzies

Anthony Mutsaers Terri O'Sullivan

Andrew Papadopoulos David Pearl

Andrew Peregrine Zvonimir Poljak

Students

Faisal Alibhai

Andreia Arruda Cathy Bauman

Lindsay Bergeron Kate Bishop-Williams

Etran Bouchouar Emily Brouwer

April Clyburne-Sherin Maryse Darch

Russell Fraser William Gow

Shawn MacKenzie Clemence Nash Peter Podobed Courtney Schott

Alexandra Swirski Krysia Walczak Lauren Wallar

Graduate Student Research Symposium 2013 ______________________________________________________________

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Graduate Studies at OVC

More information about graduate studies at the Ontario Veterinary College, University of Guelph, is available online at www.ovc.uoguelph.ca or by contacting

the graduate secretary of the department of interest:

Department of Biomedical Sciences: Wendy Arthur, Graduate Secretary

(519) 824-4120 Ext. 54900 e-mail: [email protected]

Department of Clinical Studies: Deyna Dinesen, Graduate Secretary


Department of Pathobiology: Donna Kangas, Graduate Secretary


Department of Population Medicine: Rebecca Knaggs, Graduate Secretary


More information about admission to graduate studies at the University of Guelph is available at:

http://www.ovc.uoguelph.ca/research/en/researchtrainingopportunities/futuregraduatestudents.asp


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Index of Speakers ( in alphabetical order)

Name Page Name Page Ackford, J 9 Peacock, H 31 Alibhai, F 20 Perkel, K 37 Araujo, C 24 Pieper, L 38 Arruda, A 11 Pinelli, C 37 Baquero, M 12 Pons, W 13 Barjesteh, N 18 Price, K 18 Bergeron, L 17 Quach, A 38 Berghuis, L 9 Quist, C 26 Bishop, K 28 Reilly, K 28 Bland, S 12 Remillard, L 33 Bondo, K 43 Roberts, T 14 Bujold, A 34 Roche, S 14 Diel de Amorim, M 22 Russell, K 41 Dynes, J 33 Russell, S 10 Ferris, J 35 Russell, S 40 Floras, A 22 Sargent, R 24 Fournier, A 25 Slifierz, M 39 Gallastegui, A 16 Stojsin, A 44 Gallienne, J 29 Sunohara, J 23 Garland, S 25 Swirski, A 27 Gilbert, E 42 Syed, Z 32 Gilchrist, G 36 Tang, J 10 Griffin, B 20 Tatone, E 35 Harding, S 26 Ternamian, C 31 Ho, K 23 Tessier, L 39 Ho, N 34 Tscherner, A 41 Kumagai, M 29 Tsimakouridze, E 42 Li, D 13 Weijs, C 11 Mackenzie, S 15 Wells, V 27 MacMillan, B 30 Williams, L 16 Masson, S 32 Wong, V 40 McCarty, C 36 Yu, Darrick 43 Nash, C 17 Zurbrigg, K 19 Nazarali, A 15 Oluwole,O 44 Paibomesai, M 19 Parr, J 30


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Documents

Graduate Student Research Symposium & Schofield Lecture ...€¦ · Graduate Student Research Symposium 2013 2 Introduction The tenth annual Ontario Veterinary College Graduate Student