preface - ontario veterinary college · infertility to innate immunity 2015 graduate student...

Preface

It is our great pleasure to welcome you to the 2015 Graduate Student Research

Symposium (GSRS). This volume contains the abstracts for the 94 presentations at

this year's symposium including 46 talks and 48 poster presentations. We would

like to thank the graduate students and faculty members for their participation as

well as the many sponsors of student research at the Ontario Veterinary College.

The Organizing Committee hopes that you will find this symposium to be

compelling and thought-provoking, increases awareness of research activity at the

college and provides a valuable opportunity for students and faculty to discuss

results and share ideas.

2015 GSRS Organizing Committee:

Dr. Gordon Kirby, Associate Dean, Research and Innovation

Elizabeth Lowenger, Manager, Student Affairs

Barb Gaudette, Graduate Studies and Research Assistant

Neda Barjesteh, Pathobiology

Emma Louth, Biomedical Sciences

Mehdi Emam, Pathobiology

Eamonn McGuinty, Population Medicine

Kaitlin Patterson, Population Medicine

May Kamleh, Population Medicine

Alisha Matte, Population Medicine

Emily Gilbert, Biomedical Sciences

Rashi Asthana, Clinical Studies

Carina Cooper, Clinical Studies

Shaimaa Abdelmegid, Biomedical Sciences

Professor Martin Sheldon is a veterinarian with an interest in the molecular mechanisms of infection and immunity. Professor Sheldon was in clinical practice for 14 years. He then moved to the Royal Veterinary College, where he participated in clinical teaching and developed his research interests.

Professor Sheldon completed a PhD in 2002 using a combination of clinical and laboratory studies. In 2006

he won a BBSRC Research Development Fellowship to move to full-time research and study fundamental questions about the biology of infection and immunity.

In 2008 he established a laboratory at Swansea University Medical School focussing on reproductive immunology. In 2013, Professor Sheldon’s research was recognised by the award of FRCVS.

2015 Schofield Memorial LectureWednesday, November 18

OVC Lifetime Learning Centre

Featuring:Dr. Martin Sheldon DVM, PhD

Topic: A Trail in Discovery from Animal Disease and Infertility to Innate Immunity

2015 Graduate Student Research Symposium&

Schedule of Activities:

10:00 - 11:30 AM Poster Presentations 1707 B/C12:30 - 3:30 PM Oral Presentations 1713/17153:30 - 4:30 PM Schofield Memorial Lecture 17144:30 PM Awards Ceremony & Reception 1707

GRADUATE STUDENT RESEARCH SYMPOSIUM – NOVEMBER 18, 2015

1 Goncalo Silva DVSc

A technique for determination of pulmonary artery pulse wave velocity

in horses

2 Charlotte Mitz MSc

Characterization of MAC-T cells as a New Model for the Study of TGFβ

Function in Bovine Mammary Gland Involution

3 Anuja Dharap MSc

Monitoring differences in estrous cycle length in summer and winter

seasons by measuring progesterone in water buffaloes

4 Adam Andrade MSc

Characterization of the TGFβ-TAZ Signalling Axis in Canine

Osteosarcoma Cell Lines

5 Joshua Antunes MSc

The role of microRNAs in bovine ovarian development and

angiogenesis

6 Anita Luu MSc

An investigation of the prognostic value of the TGFβ-Hippo signaling

crosstalk in canine osteosarcoma: a preliminary analysis in a tissue

microarray

7 Alham Delroba MSc

The role of the Par6-PI3K signaling axis in TGFβ tumor suppressor

function in breast cancer and its potential for therapeutic targeting

8 Casandra Merrill MSc

Mitochondrial dynamics in slow and fast growing preimplantation

bovine embryos

9 Melissa Ferguson MSc

Quitting a certainty for an uncertainty: The cardiac effects of

bisphenol S substitution for bisphenol A

10

Noeline

Subramaniam MSc

The Role of Blood Vessel Formation in Wound Healing

11 Jyoti Sharma MSc

Role of Hippo signaling pathway during bovine preimplantation

embryo development

12 Kathy Jacyniak MSc

A preliminary investigation of cardiac stem/progenitor cells in the

lizard heart (Eublepharis macularius)

13 Nina Kahlon MSc The Effects of Bisphenol A on Early Bovine Embryo Development

14 Rebecca McDonald MSc

Characterization of Neural Stem Progenitor Cells in the Brain of the

Leopard Gecko (Eublepharis macularius)

15 Joanna Smich MSc

Radiation Sensitivity of Canine Melanoma Cell Lines: Developing the

Optimal Radiotherapy Protocol

16 Georgia Kritikos MSc

Thiamine degradation in commercial pet foods stored long-term at

freezing temperatures

17 Diego Segovia BSc

Clinico-Pathological Characterization of Recently Isolated Newcastle

Disease Virus Strains in Quail

18

Wing Ka Amanda

Auyeung MSc

Mechanisms that allow oncolytic viral replication inside a tumor

despite pre-existing immunity against a virus-encoded antigen

19 Kathryn Smith MSc

Raptor mortality in Ontario, Canada: A retrospective summary (1991-

2014) and the effects of West Nile virus

20 X.T. Xie MSc

Prevalence of rotavirus, astrovirus and hepatitis E virus in Ontario

rabbit populations

21 Christine A. James MSc

Epidemiology of Ixodes scapularis on pet dogs living in an emerging

Lyme disease area in Ontario

22 Paige Vroom MSc

Microscopic Recovery of Regeneration in the Corallimorpharian,

Rhodactis sp.

23 Nahla El Skhawy MSc

T lymphocyte responses to Mycobacterium avium subspecies

paratuberculosis (MAP) -derived culture filtrate proteins and

membrane vesicles during early intestinal MAP infection

24 Mikayla Ross MSc

Bioactivity of Bovine Colostrum and Milk Exosomes and their

MicroRNA from High, Average, and Low Immune Responders on

Human Intestinal Epithelial Cells

25 Emily Arndt MSc

An investigation of the Streptococcus suis serotype distribution in

clinically ill and healthy pigs


26 Carlee Wright MSc

Contamination of Stored Drinking Water and Associations with Acute

Gastrointestinal Illness in a Canadian Inuit Community

27 Jilianne Hoffer MSc

A spatial and temporal analysis of private well water samples in

Oxford County, Canada 2005-2015

28 Emma Louth PhD

Developmental ethanol exposure disrupts attention performance and

prefrontal neuron function in male mice

29 Keith Russell PhD

Platelet lysate as alternative to fetal bovine serum in equine/canine

mesenchymal stem cell culture

30

Amir Hamed Haji

Alizadeh PhD

Characterization of clonal mesenchymal stem cell lines derived from

equine umbilical cord blood

31 Carolyn Parratt PhD MicroRNA involvement in estradiol-mediated synaptic plasticity

32 Ari Mendell PhD

The protective potential of GABAergic neurosteroids against

extracellular signal-regulated kinase-induced neurotoxicity:

preliminary findings.

33

Shaimaa

Abdelmegid PhD

Host defense-related proteins in bovine milk during subclinical

Staphylococcus aureus mastitis

34

Iman Mehdizadeh

Gohari PhD

Comparative genomic analysis of two netF-positive Clostridium

perfringens isolates associated with foal necrotizing enteritis and

canine haemorrhagic gastroenteritis

35

Megan Strachan-

Whaley PhD

Combining Oncolytic Viruses with Epigenetic Modifiers as a Novel

Therapy for Leukemia

36

Maria Rosales

Gerpe PhD

Understanding the Mechanism behind the Tissue Specificity and

Oncogenesis of JSRV amd ENTV in sheep

37 Joelle Ingrao PhD

Development of orf virus (parapoxvirus ovis) as a multivalent viral

vector platform against Toxoplasma gondii

38 Laura Bassel PhD

Identification of Bordetella bronchiseptica in fatal pneumonia of dogs

and cats

39

Rubee

Bavananthasivam PhD

Induction of innate responses in chicken embryos by administration of

encapsulated and free forms of a Toll-like receptor 4 ligand

40 Maureen Jarau PhD

Virulence diversity of Flavobacterium psychrophilum isolates in

Ontario and correlation of bacterial numbers in fish spleen with

mortality

41 Abbie Viscardi PhD

Pharmacokinetics and potential toxicity of high dose meloxicam in

preweaned piglets

42 Rob Mould PhD

Combining virotherapy with immunotherapy to treat osteosarcoma in

a preclinical and clinical model

43

Kathy

Matuszewska PhD

Combined Vessel Normalization and Oncolytic Virus Therapy in the

Treatment of Advanced Stage Ovarian Cancer

44 Kristin Bondo PhD

Epidemiology of Salmonella in Raccoons and their Environment on

Swine Farms and Conservation Areas

45 Stephanie Hughes PhD

Augmenting surveillance to minimize the burden of norovirus-like

illness in Ontario: Using TeleHealth Ontario data to detect the onset of

community activity

46 Emma Gardner PhD

Using an Agent-Based Model to Compare Two Diagnostic Tests for

Staphylococcus aureus Bovine Mastitis

47 Amanda Perri PhD

A comparison of using beta-hydroxybutyrate versus clinical signs to

identify anorexic nursery pigs

48 Alexandra Swirski PhD

A comparison of exposure to risk factors for giardiasis among non-

travelers, domestic travelers and international travelers in a Canadian

community (2006-2012)


12:30-13:22 ORAL PRESENTATIONS I

Room 1713 LLC Room 1715 LLC Room 1810 PATH Room 1812 PATH

12:30-12:43 1

Danielle Hopkins (MSc)

Investigation of the potential relationship between swine

influenza virus and Streptococcus suis infections in weanling pigs

Cristine Reitz (PhD)

CLOCK Coordinates T Cell Recruitment and Proliferation to

Benefit Outcome Post-Myocardial Infarction

Benoît Cuq (DVSc)

Reproducibility, Stability and Reference Ranges of Thrombin Generation using the Calibrated Automated Thrombogram in a

Population of Health Dogs

Russell S. Fraser (PhD)

Investigating infectious disease susceptibility in horses through

targeted next-generation resequencing of the collagenous

lectin gene family

12:43-12:56 2

Saranya Nair (MSc)

A Longitudinal Study of the Influence of Flavophospholipol on

Salmonella Shedding and Colonization in Naturally Infected

Pigs

Faisal Alibhai (PhD)

Disrupting the Key Circadian Regulator CLOCK leads to Age-

Dependent Heart Disease

Jacqueline Scott (DVSc)

Perioperative outcome and complications following

laparoscopic cholecystectomy in dogs: 20 cases (2008-2015)

Laurence Tessier (PhD)

RNA-seq profiling of the bronchiolar epithelium of horses

with recurrent airway obstruction: insights into pathologic response

to tissue damage in the lung

12:56-13:09 3

Craig Leroy (MSc)

Accuracy and intensity of estrus detection with activity monitoring systems for lactating dairy cows

Elena Tsimakouridze (PhD)

The Primary Benefits of Angiotensin-Converting Enzyme

Inhibition on Cardiac Remodeling Occur During Sleep Time in

Murine Myocardial Ischemia-Reperfusion (heart attack)

Guillaume Leblond (DVSc)

Biomechanical and Radiological Comparison between

Transarticular Screw Fixation and 2 Multi-Implant Constructs for

Ventral Atlantoaxial Stabilization in the Dog.

Diego Gomez (PhD)

Characterization of the fecal bacterial microbiota of healthy

and diarrheic calves

13:09-13:22 4

Beryl Chung (PhD)

The α4β2* nicotinic receptor and the developing hippocampal

formation

Cyrielle Finck (DVSc)

Structural and Functional Integrity of Microwave Ablation Antennas

with Repeated Use and Re-sterilization.

Kevin Stinson (PhD)

Evidence for early pathogen clearance in a subclinical Johne's

disease calf model

13:22-13:32 BREAK


13:32-14:24 ORAL PRESENTATIONS II

Room 1713 Room 1715 Room 1810 Room 1812

13:32-13:45 1

Matthew Little (PhD)

Factors associated with glucose tolerance, pre-diabetes, and type 2

diabetes in a rural community of south India: A cross-sectional study

Sarah Lepage (PhD)

Cartilage tissue engineering using equine cord blood-derived mesenchymal stromal cells

Delphine Laniesse (DVSc)

Rheological properties of a sustained-release formulation of butorphanol in a poloxamer 407

base and pharmacokinetics in Amazon parrots (Amazona ventralis)

Jacob van Vloten (PhD)

Harnessing immunogenic cell death to potentiate anti-cancer efficacy during ORFV-induced

oncolysis.

13:45-13:58 2

Kaitlin Patterson (PhD)

Seasonal variation of food security among the Batwa of Kanungu

District, Uganda

Kelsey Spence (PhD)

Characterization of the equine contact network at a single

equestrian show

Laura Bourque (DVSc)

Environmental Toxicants and Innate Pulmonary Defence: The

Effects of Polycylic Aromatic Hydrocarbons on in vitro

Expression of Beta Defensins in Tracheal Epithelial Cells

Katie Clow (PhD)

Distribution of ticks and the risk of Lyme disease and other tick-

borne pathogens of public health significance in Ontario, Canada

13:58-14:11 3

Laura Jane Weber (PhD)

Health-Seeking Behaviour in Response to Acute Gastrointestinal Illness among the Ugandan Batwa

Tanya Rossi (PhD)

Post-Exercise Cardiac Troponin I Release and Clearance in Normal

Standardbred Racehorses

Jennifer Kylie (DVSc)

Factors Affecting the Composition of the Rabbit Fecal Microbiota

Mehdi Emam (PhD)

The effect of immunological status, in-vitro treatment and culture time on expression of

eleven candidate reference genes in bovine blood mononuclear cells

14:11-14:24 4

Monica Baquero (PhD)

WC1+ γδ T lymphocytes from calves and heifers differentially modulate macrophage function

during early Mycobacterium avium subspecied

paratuberculosis infection.

Nariman Deravi (DVSc)

Prognostication of Canine T-cell Lymphoma

Alex Leveille (PhD)

An undescribed species of Hepatozoon infecting the blood of

northern leopard frogs in Southern Ontario is uncovered using mitochondrial genome

targets.

14:24-14:34 BREAK


14:34-15:26 ORAL PRESENTATIONS III

Room 1713 Room 1715 Room 1810 Room 1812

14:34-14:47 1

Christopher Pinelli (PhD)

Timp3 loss confers androgen-independence in a syngeneic mouse

model of castration-resistant prostate cancer

Amanda Macdonald (PhD)

Disease Assessment of Wild Turkeys (Meleagris gallopavo) in

Ontario

Charlotte Winder (DVSc)

Practices for disbudding and dehorning of dairy calves by

veterinarians and dairy producers in Ontario, Canada

Jessica Kilkenny (MSc)

Evaluation of Laparoscopic Skills in Doctor of Veterinary Medicine

Students

14:47-15:00 2

Courtney Schott (PhD)

Expression Patterns of Autophagy-Associated Proteins in Canine Appendicular Osteosarcoma

Alexandra Sawatzky (PhD)

Exploring pathways for achieving and sustaining good wellbeing

among Inuit in Nunatsiavut, Canada

Firdous Khan (DVSc)

Nitric oxide’s dose-dependent inhibition of uterine contractility: a

potential mechanism underlying persistent breeding-induced

endometritis in the mare

Rebecca Flancman (MSc)

Evaluation of the Impact of a Dental Prophylaxis on the Oral

Microbiota of Canines

15:00-15:13 3

Hannah Flint (PhD)

Risk factors associated with stranger-directed aggression in dogs

Elise Tatone (PhD)

A systematic review and meta-analysis of on-farm methods for

the detection of hyperketonemia in dairy cattle

Juliana Bonin Ferreira (DVSc)

Swine influenza - patterns and determinants of virus transmission

in a nursery herd

Jessica Reimann (MSc)

Perioperative analgesic use by Ontario veterinarians, 2012

15:13-15:26 4

Lauren Dawson (PhD)

Pain assessment and management in cats and dogs by veterinarians in

southwestern Ontario

Andreia Arruda (PhD)

The use of network analysis for investigation of risk factors for

porcine reproductive and respiratory syndrome (PRRS)

15:30-16:30

LLC 1714

SCHOFIELD LECTURE:

“A trail in discovery from animal disease and infertility to innate immunity”

Dr. Martin Sheldon DVM, PhD

16:30-17:00 RECEPTION & PRESENTATION OF AWARDS

Table of Contents

I

EVALUATION OF LAPAROSCOPIC SKILLS IN VETERINARY STUDENTS ____________________________ 1

KILKENNY, J1., SINGH, A1., KERR, C1., KHOSA, D2., AND FRANSSON, B3.

DEVELOPMENTAL ETHANOL EXPOSURE DISRUPTS ATTENTION PERFORMANCE AND PREFRONTAL

NEURON FUNCTION IN MALE MICE ______________________________________________________ 2

LOUTH, E., BIGNELL, W., TAYLOR, C. AND BAILEY, C.

DISTRIBUTION OF TICKS AND THE RISK OF LYME DISEASE AND OTHER TICK-BORNE PATHOGENS OF

PUBLIC HEALTH SIGNIFICANCE IN ONTARIO, CANADA _______________________________________ 3

CLOW, K.M., OGDEN, N. H., LINDSAY, L. R., MICHEL, P., PEARL, D., AND JARDINE, C. M.

EPIDEMIOLOGY OF SALMONELLA IN RACCOONS AND THEIR ENVIRONMENT ON SWINE FARMS AND

CONSERVATION AREAS _______________________________________________________________ 4

BONDO, K. J., PEARL, D. L., JANECKO, N., BOERLIN, P., REID-SMITH, R. J., PARMLEY, J., AND JARDINE, C. M.

AUGMENTING SURVEILLANCE TO MINIMIZE THE BURDEN OF NOROVIRUS-LIKE ILLNESS IN ONTARIO:

USING TELEHEALTH ONTARIO DATA TO DETECT THE ONSET OF COMMUNITY ACTIVITY ____________ 5

HUGHES, S. L., AND PAPADOPOULOS, A.

PRACTICES FOR DISBUDDING AND DEHORNING OF DAIRY CALVES BY VETERINARIANS AND DAIRY

PRODUCERS IN ONTARIO, CANADA _____________________________________________________ 6

WINDER, C., LEBLANC, S., HALEY, D., LISSEMORE, K., GODKIN, K. AND DUFFIELD, T.

PERIOPERATIVE OUTCOME AND COMPLICATIONS FOLLOWING LAPAROSCOPIC CHOLECYSTECTOMY IN

DOGS: 20 CASES (2008-2015) ___________________________________________________________ 7

SCOTT, J.1, SINGH, A.1, MAYHEW, P. D.2, CASE, J. B.3, RUNGE, J. J.4, AND GATINEAU, M.5

A SYSTEMATIC REVIEW AND META-ANALYSIS OF ON-FARM METHODS FOR THE DETECTION OF

HYPERKETONEMIA IN DAIRY CATTLE ____________________________________________________ 8

TATONE, E.1*, GORDON, J. 1, HUBBS, J.1, DEVRIES, T.2, LEBLANC, S.1, AND DUFFIELD, T.1

CHARACTERIZATION OF MAC-T CELLS AS A NEW MODEL FOR THE STUDY OF TGFΒ FUNCTION IN

BOVINE MAMMARY GLAND INVOLUTION ________________________________________________ 9

MITZ, C., AND VILORIA-PETIT, A.

II

BIOMECHANICAL AND RADIOLOGICAL COMPARISON BETWEEN TRANSARTICULAR SCREW FIXATION

AND 2 MULTI-IMPLANT CONSTRUCTS FOR VENTRAL ATLANTOAXIAL STABILIZATION IN THE DOG __ 10

LEBLOND, G.1, GAITERO, L.1, MOENS, N. M. M.1, LINDEN, A. Z.1, JAMES, F. M. K1, MONTEITH, G.1, AND

RUNCIMAN, R. J.2

COMPARATIVE GENOMIC ANALYSIS OF TWO NETF-POSITIVE CLOSTRIDIUM PERFRINGENS ISOLATES

ASSOCIATED WITH FOAL NECROTIZING ENTERITIS AND CANINE HAEMORRHAGIC GASTROENTERITIS.11

MEHDIZADEH GOHARI, I.1, KROPINSKI, A.1, WEESE, S.1, PARREIRA, V.1, WHITEHEAD, A.2, BOERLIN, P.1, AND

PRESCOTT, J.1

THE PRIMARY BENEFITS OF ANGIOTENSIN-CONVERTING ENZYME INHIBITION ON CARDIAC

REMODELING OCCUR DURING SLEEP TIME IN MURINE MYOCARDIAL ISCHEMIA-REPERFUSION (HEART

ATTACK) __________________________________________________________________________ 12

TSIMAKOURIDZE, E., ALIBHAI, F., REITZ, C., AND MARTINO, T.

THE EFFECT OF IMMUNOLOGICAL STATUS, IN-VITRO TREATMENT AND CULTURE TIME ON EXPRESSION

OF ELEVEN CANDIDATE REFERENCE GENES IN BOVINE BLOOD MONONUCLEAR CELLS ____________ 13

EMAM, M., THOMPSON-CRISPI, K., AND MALLARD, B.

CHARACTERIZATION OF THE EQUINE CONTACT NETWORK AT A SINGLE EQUESTRIAN SHOW ______ 14

SPENCE, K.L., GOH, B., O’SULLIVAN, T.L., POLJAK, Z., AND GREER, A.L.

DEPARTMENT OF POPULATION MEDICINE

REPRODUCIBILITY, STABILITY AND REFERENCE RANGES OF THROMBIN GENERATION USING THE

CALIBRATED AUTOMATED THROMBOGRAM IN A POPULATION OF HEALTHY DOGS. _____________ 15

CUQ, B.1, BLOIS, S.1, WOOD, D.2, ABRAMS-OGG, A.1 AND BÉDARD, C.3

STRUCTURAL AND FUNCTIONAL INTEGRITY OF MICROWAVE ABLATION ANTENNAS WITH REPEATED

USE AND RE-STERILIZATION ___________________________________________________________ 16

FINCK, C., ZUR LINDEN, A., SINGH, A., FOSTER, R. AND NYKAMP, S.

COMBINING ONCOLYTIC VIRUSES WITH EPIGENETIC MODIFIERS AS A NOVEL THERAPY FOR LEUKEMIA

_________________________________________________________________________________ 17

STRACHAN-WHALEY M, AU YEUNG A, KIM J, BIENZLE D, MUTSAERS A, WOOTTON S, BRIDLE B

FACTORS AFFECTING THE COMPOSITION OF THE RABBIT FECAL MICROBIOTA __________________ 18

KYLIE, J., WEESE, J. S. AND TURNER, P.V.

III

POST-EXERCISE CARDIAC TROPONIN I RELEASE AND CLEARANCE IN NORMAL STANDARDBRED

RACEHORSES _______________________________________________________________________ 19

ROSSI, T. M.1, PEARL, D. L.1, PYLE, W. G.2, MAXIE, M. G.3, KAVSAK, P. A.4, AND PHYSICK-SHEARD, P. W.1

UNDERSTANDING THE MECHANISM BEHIND THE TISSUE SPECIFICITY AND ONCOGENESIS OF JSRV AND

ENTV IN SHEEP _____________________________________________________________________ 20

ROSALES GERPE, M. R.1, WALSH, S. R.2, DE JONG, J.1, AND WOOTTON, S. K.1

MONITORING DIFFERENCES IN ESTROUS CYCLE LENGTH IN SUMMER AND WINTER SEASONS BY

MEASURING PROGESTERONE IN WATER BUFFALOES ______________________________________ 21

DHARAP, A.1, GILL, I.1, MORADI, F.1, ASHKAR, F. A.1, ROBERTSON, L.1, WEST, C.2, LITTKEMANN, M.2, SMITH, L.2,

ANTENOS, M.1, AND KING, W. A.1

CHARACTERIZATION OF THE TGFΒ-TAZ SIGNALLING AXIS IN CANINE OSTEOSARCOMA CELL LINES __ 22

ANDRADE, A. D.1, NEOGI, P.1, MUTSAERS, T.1, WOOD, G.2, AND VILORIA-PETIT, A.1

HEALTH-SEEKING BEHAVIOUR IN RESPONSE TO ACUTE GASTROINTESTINAL ILLNESS AMONG THE

UGANDAN BATWA __________________________________________________________________ 23

WEBER, L.J. AND INDIGENOUS HEALTH ADAPTATION TO CLIMATE CHANGE (IHACC) RESEARCH TEAM

A LONGITUDINAL STUDY OF THE INFLUENCE OF FLAVOPHOSPHOLIPOL ON SALMONELLA SHEDDING

AND COLONIZATION IN NATURALLY INFECTED PIGS _______________________________________ 24

NAIR, S., FARZAN, A., O’SULLIVAN, T., AND FRIENDSHIP, R.M.

RHEOLOGICAL PROPERTIES OF A SUSTAINED-RELEASE FORMULATION OF BUTORPHANOL IN A

POLOXAMER 407 BASE AND PHARMACOKINETICS IN AMAZON PARROTS (AMAZONA VENTRALIS)__ 25

LANIESSE, D.¹, SANCHEZ-MIGALLON GUZMAN², D., SMITH, D.¹, KNYCH, H.², MOSLEY, C.¹, PAUL-MURPHY, J.², AND

BEAUFRÈRE, H.¹

T LYMPHOCYTE RESPONSES TO MYCOBACTERIUM AVIUM SUBSPECIES PARATUBERCULOSIS (MAP) -

DERIVED CULTURE FILTRATE PROTEINS AND MEMBRANE VESICLES DURING EARLY INTESTINAL MAP

INFECTION_________________________________________________________________________ 26

EL SKHAWY, N., AND PLATTNER, N.

DEVELOPMENT OF ORF VIRUS (PARAPOXVIRUS OVIS) AS A MULTIVALENT VIRAL VECTOR PLATFORM

AGAINST TOXOPLASMA GONDII _______________________________________________________ 27

INGRAO, J.C., VAN VLOTEN, J.P., SHAPIRO, K., BARTA, J.R., MENZIES, P.I., BRIDLE, B.W., AND WOOTTON, S.K.

IV

HARNESSING IMMUNOGENIC CELL DEATH TO POTENTIATE ANTI-CANCER EFFICACY DURING ORFV-

INDUCED ONCOLYSIS. _______________________________________________________________ 28

VAN VLOTEN, J. P., BRIDLE, B.W., AND WOOTTON, S.K.

ACCURACY AND INTENSITY OF ESTRUS DETECTION WITH ACTIVITY MONITORING SYSTEMS FOR

LACTATING DAIRY COWS _____________________________________________________________ 29

LEROY, C. AND LEBLANC, S.J.

USING AN AGENT-BASED MODEL TO COMPARE TWO DIAGNOSTIC TESTS FOR STAPHYLOCOCCUS

AUREUS BOVINE MASTITIS ___________________________________________________________ 30

GARDNER, E.1, KELTON, D.1, HAND, K.2, POLJAK, Z.1, AND GREER, A. L.1

PROGNOSTICATION OF CANINE T-CELL LYMPHOMA _______________________________________ 31

DERAVI, N.1, BERKE, O.2, BIENZLE, D.1

IDENTIFICATION OF BORDETELLA BRONCHISEPTICA IN FATAL PNEUMONIA OF DOGS AND CATS ___ 32

BASSEL, L.L.¹, ABDELAZIZ, K.¹², HARNESS, M.L.¹, CLARK, M.E.¹, REGISTER, K.B.³, CASWELL, J.L.¹

PLATELET LYSATE AS ALTERNATIVE TO FETAL BOVINE SERUM IN EQUINE/CANINE MESENCHYMAL

STEM CELL CULTURE _________________________________________________________________ 33

RUSSELL, K. A., GIBSON, T. W., CHONG, A., CO, C., AND KOCH, T. G.

A COMPARISON OF USING BETA-HYDROXYBUTYRATE VERSUS CLINICAL SIGNS TO IDENTIFY ANOREXIC

NURSERY PIGS _____________________________________________________________________ 34

PERRI AM1, FRIENDSHIP R1, HARDING JCS2, O’SULLIVAN TL1

MECHANISMS THAT ALLOW ONCOLYTIC VIRAL REPLICATION INSIDE A TUMOR DESPITE PRE-EXISTING

IMMUNITY AGAINST A VIRUS-ENCODED ANTIGEN ________________________________________ 35

AUYEUNG, W. K. A., MOULD, R. C., SPANGLER, H., KIM, J., AND W. BRIDLE, B. W.

AN UNDESCRIBED SPECIES OF HEPATOZOON INFECTING THE BLOOD OF NORTHERN LEOPARD FROGS

IN SOUTHERN ONTARIO IS UNCOVERED USING MITOCHONDRIAL GENOME TARGETS. ___________ 36

LEVEILLE, A. N., AND BARTA, J. R.

RAPTOR MORTALITY IN ONTARIO, CANADA: A RETROSPECTIVE SUMMARY (1991-2014) AND THE

EFFECTS OF WEST NILE VIRUS _________________________________________________________ 37

SMITH, K., CAMPBELL, D., PEARL, D., SALGADO-BIERMAN, F. AND NEMETH, N.

V

PREVALENCE OF ROTAVIRUS, ASTROVIRUS AND HEPATITIS E VIRUS IN ONTARIO RABBIT

POPULATIONS ______________________________________________________________________ 38

XIE, X. T., BIL, J. HAMMERMUELLER, J., AND TURNER, P.V.

THIAMINE DEGRADATION IN COMMERCIAL PET FOODS STORED LONG-TERM AT FREEZING

TEMPERATURES ____________________________________________________________________ 39

KRITIKOS, G, PARR, J, MARTIN, B, AND VERBRUGGHE, A

NITRIC OXIDE’S DOSE-DEPENDENT INHIBITION OF UTERINE CONTRACTILITY: A POTENTIAL

MECHANISM UNDERLYING PERSISTENT BREEDING-INDUCED ENDOMETRITIS IN THE MARE _______ 40

KHAN, F.A.1, CHENIER, T.S.1, MURRANT, C.L.2, FOSTER, R.A.3, HEWSON, J.4, AND SCHOLTZ, E.L.1

THE ROLE OF MICRORNAS IN BOVINE OVARIAN DEVELOPMENT AND ANGIOGENESIS ____________ 41

ANTUNES, J. AND PETRIK, J.

DISEASE ASSESSMENT OF WILD TURKEYS (MELEAGRIS GALLOPAVO) IN ONTARIO _______________ 42

MACDONALD, A. M., REJMAN, E., BARTA, J., AND NEMETH, N.

EXPLORING PATHWAYS FOR ACHIEVING AND SUSTAINING GOOD WELLBEING AMONG INUIT IN

NUNATSIAVUT, CANADA _____________________________________________________________ 43

SAWATZKY, A.1, CUNSOLO WILLOX, A.2, HARPER, S.1, SHIWAK, I.3, WOOD, M.4, FORD, J.5, THE INUIT MENTAL

HEALTH AND ADAPTATION TO CLIMATE CHANGE TEAM, AND THE RIGOLET INUIT COMMUNITY GOVERNMENT 6

AN INVESTIGATION OF THE PROGNOSTIC VALUE OF THE TGFΒ-HIPPO SIGNALING CROSSTALK IN

CANINE OSTEOSARCOMA: A PRELIMINARY ANALYSIS IN A TISSUE MICROARRAY ________________ 44

LUU, A1., SCHOTT, C.2, MUTSAERS, A.1,WOOD, G.2,VILORIA-PETIT, A.M1.

CARTILAGE TISSUE ENGINEERING USING EQUINE CORD BLOOD-DERIVED MESENCHYMAL STROMAL

CELLS _____________________________________________________________________________ 45

LEPAGE, S., KANDEL, R., AND KOCH, T.

THE ROLE OF THE PAR6- PI3K SIGNALING AXIS IN TGFΒ TUMOR SUPPRESSOR FUNCTION IN BREAST

CANCER AND ITS POTENTIAL FOR THERAPEUTIC TARGETING ________________________________ 46

DELROBA, A. AND VILORIA-PETIT, A. M.

EPIDEMIOLOGY OF IXODES SCAPULARIS ON PET DOGS LIVING IN AN EMERGING LYME DISEASE AREA

IN ONTARIO _______________________________________________________________________ 47

JAMES, C. A., PEARL, D. L., PEREGRINE, A. S., LINDSAY, L. R., AND JARDINE, C. M.

VI

A COMPARISON OF EXPOSURE TO RISK FACTORS FOR GIARDIASIS AMONG NON-TRAVELERS,

DOMESTIC TRAVELERS AND INTERNATIONAL TRAVELERS IN A CANADIAN COMMUNITY (2006-2012) 48

SWIRSKI, A., PEARL, D., PEREGRINE, A. AND PINTAR, K.

ENVIRONMENTAL TOXICANTS AND INNATE PULMONARY DEFENCE: THE EFFECTS OF POLYCYCLIC

AROMATIC HYDROCARBONS ON IN VITRO EXPRESSION OF BETA DEFENSINSIN TRACHEAL EPITHELIAL

CELLS _____________________________________________________________________________ 49

BOURQUE, L., RAVERTY, S., B. LILLIE AND CASWELL, J.

TIMP3 LOSS CONFERS ANDROGEN-INDEPENDENCE IN A SYNGENEIC MOUSE MODEL OF CASTRATION-

RESISTANT PROSTATE CANCER ________________________________________________________ 50

PINELLI, CJ., KIM, SD. AND WOOD, GA.

MICROSCOPIC RECOVERY OF REGENERATION IN THE CORALLIMORPHARIAN, RHODACTIS SP. _____ 51

VROOM, P., PETERS, E. AND LUMSDEN, J.

EVALUATION OF THE IMPACT OF A DENTAL PROPHYLAXIS ON THE ORAL MICROBIOTA OF CANINES 52

FLANCMAN, R. AND WEESE, JS.

CHARACTERIZATION OF CLONAL MESENCHYMAL STEM CELL LINES DERIVED FROM EQUINE UMBILICAL

CORD BLOOD ______________________________________________________________________ 53

HAJI ALIZADEH, AH. AND KOCH, T.

THE USE OF NETWORK ANALYSIS FOR INVESTIGATION OF RISK FACTORS FOR PORCINE REPRODUCTIVE

AND RESPIRATORY SYNDROME (PRRS) __________________________________________________ 54

ARRUDA, A G.1, FRIENDSHIP, R.1, CARPENTER, J.2, HAND, K.3 AND POLJAK, Z.1

RNA-SEQ PROFILING OF THE BRONCHIOLAR EPITHELIUM OF HORSES WITH RECURRENT AIRWAY

OBSTRUCTION: INSIGHTS INTO PATHOLOGIC RESPONSE TO TISSUE DAMAGE IN THE LUNG _______ 55

TESSIER, L., CÔTÉ, O., VIEL, L., CLARK, ME., DIAZ, A., ANDERS, S. AND BIENZLE, D .

FACTORS ASSOCIATED WITH GLUCOSE TOLERANCE, PRE-DIABETES, AND TYPE 2 DIABETES IN A RURAL

COMMUNITY OF SOUTH INDIA: A CROSS-SECTIONAL STUDY ________________________________ 56

LITTLE, M.1, HUMPHRIES, S.2, PATEL, K.3 AND DEWEY C1

WC1+ ΓΔ T LYMPHOCYTES FROM CALVES AND HEIFERS DIFFERENTIALLY MODULATE MACROPHAGE

FUNCTION DURING EARLY MYCOBACTERIUM AVIUM SUBSPECIED PARATUBERCULOSIS INFECTION. 57

BAQUERO, M. AND PLATTNER, B.

VII

TRENDS IN CAT ADMISSIONS TO GUELPH ANIMAL SHELTER BETWEEN 2011 AND 2015 ___________ 58

JANKE, N., BERKE, O., KLEMENT, E., COE, J. AND FLOCKHART, T.

CLINICO-PATHOLOGICAL CHARACTERIZATION OF RECENTLY ISOLATED NEWCASTLE DISEASE VIRUS

STRAINS IN QUAIL ISSING ____________________________________________________________ 59

SEGOVIA, D., MILLER, P. AND SUSTA, L.

RISK FACTORS ASSOCIATED WITH STRANGER-DIRECTED AGGRESSION IN DOGS _________________ 60

FLINT, H.1, COE, J.1, SERPELL, J.2, PEARL, DL.1 AND NIEL, L.1

PERIOPERATIVE ANALGESIC USE BY ONTARIO VETERINARIANS, 2012 _________________________ 61

REIMANN, J., DEWEY, C., BATEMAN, S., KERR, C. AND JOHNSON, R.

INDUCTION OF INNATE RESPONSES IN CHICKEN EMBRYOS BY ADMINISTRATION OF ENCAPSULATED

AND FREE FORMS OF A TOLL-LIKE RECEPTOR 4 LIGAND ____________________________________ 62

BAVANANTHASIVAM, J., ALKIE, T., HODGINS, DC. AND SHARIF, S.

AN INVESTIGATION OF THE STREPTOCOCCUS SUIS SEROTYPE DISTRIBUTION IN CLINICALLY ILL AND

HEALTHY PIGS ______________________________________________________________________ 63

ARNDT, E.1, FARZAN, V.1, SOLTES, G.2, MACINNES, J.2 AND FRIENDSHIP, R.1

MITOCHONDRIAL DYNAMICS IN SLOW AND FAST GROWING PREIMPLANTATION BOVINE EMBRYOS 64

MERRILL, C., PERKEL, K. AND MADAN, P.

CHARACTERIZATION OF THE FECAL BACTERIAL MICROBIOTA OF HEALTHY AND DIARRHEIC CALVES. 65

GOMEZ, DE.1, ARROYO, LG.2, VIEL, L.2 AND WEESE, JS1.

METEOROLOGICAL DRIVERS OF ACUTE RESPIRATORY INFECTIONS IN RURAL SOUTHWESTERN

UGANDA __________________________________________________________________________ 66

BISHOP-WILLIAMS, K.1, SARGEANT, J.1, BERRANG-FORD, L.2, EDGE, .V1, IHACC RESEARCH GROUP3 AND HARPER,

S.1

BIOACTIVITY OF BOVINE COLOSTRUM AND MILK EXOSOMES AND THEIR MICRORNA FROM HIGH,

AVERAGE, AND LOW IMMUNE RESPONDERS ON HUMAN INTESTINAL EPITHELIAL CELLS __________ 67

ROSS, M.1, ATALLA, H.1, 2, GALLO, N.1, SYJUECO, E.1 AND MALLARD, B.1, 2

VIII

VIRULENCE DIVERSITY OF FLAVOBACTERIUM PSYCHROPHILUM ISOLATES IN ONTARIO AND

CORRELATION OF BACTERIAL NUMBERS IN FISH SPLEEN WITH MORTALITY ____________________ 68

JARAU, M., DI NATALE, A., HUBER, P., MACINNES, J. AND LUMSDEN, J.

PHARMACOKINETICS AND POTENTIAL TOXICITY OF HIGH DOSE MELOXICAM IN PREWEANED PIGLETS

_________________________________________________________________________________ 69

VISCARDI, A.1, COETZEE, J.2 AND TURNER, PV.1

THE Α4Β2* NICOTINIC RECEPTOR AND THE DEVELOPING HIPPOCAMPAL FORMATION ___________ 70

CHUNG, B. AND BAILEY, C.

INVESTIGATING INFECTIOUS DISEASE SUSCEPTIBILITY IN HORSES THROUGH TARGETED NEXT-

GENERATION RESEQUENCING OF THE COLLAGENOUS LECTIN GENE FAMILY ____________________ 71

FRASER, RS., HAMMERMUELLER, JD., MEYER, A. AND LILLIE, BN.

MICRORNA INVOLVEMENT IN ESTRADIOL-MEDIATED SYNAPTIC PLASTICITY ___________________ 72

PARRATT, C., LAMARRE, J. AND MACLUSKY, N.

QUITTING A CERTAINTY FOR AN UNCERTAINTY: THE CARDIAC EFFECTS OF BISPHENOL S SUBSTITUTION

FOR BISPHENOL A ___________________________________________________________________ 73

FERGUSON, M. AND PYLE, WG.

THE ROLE OF BLOOD VESSEL FORMATION IN WOUND HEALING______________________________ 74

SUBRAMANIAM, N., PETRIK, J. AND VICKARYOUS, M.

ROLE OF HIPPO SIGNALING PATHWAY DURING BOVINE PREIMPLANTATION EMBRYO DEVELOPMENT

_________________________________________________________________________________ 75

SHARMA, J. AND MADAN, P.

A PRELIMINARY INVESTIGATION OF CARDIAC STEM/PROGENITOR CELLS IN THE LIZARD HEART

(EUBLEPHARIS MACULARIUS) _________________________________________________________ 76

JACYNIAK, K., HAWTHORNE, D., ROBERTSON, L. AND VICKARYOUS, M.

THE EFFECTS OF BISPHENOL A ON EARLY BOVINE EMBRYO DEVELOPMENT ____________________ 77

KAHLON, N., FERRIS, JC., MACLUSKY, N., KING, WA. AND FAVETTA, LA.

IX

CONTAMINATION OF STORED DRINKING WATER AND ASSOCIATIONS WITH ACUTE

GASTROINTESTINAL ILLNESS IN A CANADIAN INUIT COMMUNITY ____________________________ 78

WRIGHT, C.1, SHIWAK, I.2, FORD, J.3, FARAHBAKHSH, K.1, EDGE, V.1, RICG3, IHACC RESEARCH GROUP4 AND

HARPER, SL.1

INVESTIGATION OF THE POTENTIAL RELATIONSHIP BETWEEN SWINE INFLUENZA VIRUS AND

STREPTOCOCCUS SUIS INFECTIONS IN WEANLING PIGS _____________________________________ 79

HOPKINS, D., POLJAK, Z., FARZAN, V., GRGIĆ, H. AND FRIENDSHIP, R.

PAIN ASSESSMENT AND MANAGEMENT IN CATS AND DOGS BY VETERINARIANS IN SOUTHWESTERN

ONTARIO __________________________________________________________________________ 80

DAWSON, L.1, DEWEY, C.1, STONE, E.2, GUERIN, M.1, MOSLEY, C.3 AND NIEL, L.1

EVIDENCE FOR EARLY PATHOGEN CLEARANCE IN A SUBCLINICAL JOHNE’S DISEASE CALF MODEL. __ 81

STINSON, KJ. AND PLATTNER, B.

CHARACTERIZATION OF NEURAL STEM/PROGENITOR CELLS IN THE BRAIN OF THE LEOPARD GECKO

(EUBLEPHARIS MACULARIUS) _________________________________________________________ 82

MACDONALD, A., MCDONALD, R., GILBERT, E. AND VICKARYOUS, MK.

CLOCK COORDINATES T CELL RECRUITMENT AND PROLIFERATION TO BENEFIT OUTCOME POST-

MYOCARDIAL INFARCTION ___________________________________________________________ 83

REITZ, C., TSIMAKOURIDZE, E., ALIBHAI, F. AND MARTINO, T.

THE PROTECTIVE POTENTIAL OF GABAERGIC NEUROSTEROIDS AGAINST EXTRACELLULAR SIGNAL-

REGULATED KINASE-INDUCED NEUROTOXICITY: PRELIMINARY FINDINGS ______________________ 84

MENDELL, A. AND MACLUSKY, N.

A TECHNIQUE FOR DETERMINATION OF PULMONARY ARTERY PULSE WAVE VELOCITY IN HORSES__ 85

SILVA, G.1, GUEST, B.2, GOMEZ, DE.1, MCGREGOR, M.2, RUNCIMAN, J.2AND ARROYO, LG.1

COMBINING VIROTHERAPY WITH IMMUNOTHERAPY TO TREAT OSTEOSARCOMA IN A PRECLINICAL

AND CLINICAL MODEL _______________________________________________________________ 86

MOULD, R., KIM, J., WALSH, S., DE JONG, J., WOOD, G., WOOTTON, S., SUSTA, L., PETRIK, J., MUSTAERS, T., AND

BRIDLE, B.

HOST DEFENSE-RELATED PROTEINS IN BOVINE MILK DURING SUBCLINICAL STAPHYLOCOCCUS AUREUS

MASTITIS __________________________________________________________________________ 87

ABDELMEGID, S.1, PARK, A.2, ATALLA, H.3, MACKAY, A.1, CASWELL, J.3, KELTON, D.4 , KIRBY, G.1

X

SEASONAL VARIATION OF FOOD SECURITY AMONG THE BATWA OF KANUNGU, UGANDA ________ 88

PATTERSON, K., BERRANG-FORD, L., LWASA, S., NAMANYA, D., FORD, J., TWEBAZE, F., CLARK, S., DONNELLY, B.,

IHACC RESEARCH TEAM, HARPER, S.

A SPATIAL AND TEMPORAL ANALYSIS OF PRIVATE WELL WATER SAMPLES IN OXFORD COUNTY,

CANADA 2005-2015 _________________________________________________________________ 89

HOFFER, J., BERKE, O. AND ALTON, G.

DISRUPTING THE KEY CIRCADIAN REGULATOR CLOCK LEADS TO AGE-DEPENDENT HEART DISEASE _ 90

ALIBHAI, F., TSIMAKOURIDZE, E., LAMARRE, J. AND MARTINO, T.

EXPRESSION PATTERNS OF AUTOPHAGY-ASSOCIATED PROTEINS IN CANINE APPENDICULAR

OSTEOSARCOMA ___________________________________________________________________ 91

SCHOTT, CR. AND WOOD, GA.

SWINE INFLUENZA - PATTERNS AND DETERMINANTS OF VIRUS TRANSMISSION IN A NURSERY HERD 92

FERREIRA, JB., GRGIĆ, H., FRIENDSHIP, R., WIDEMAN, G., NAGY, Ѐ. AND POLJAK, Z.

RADIATION SENSITIVITY OF CANINE MELANOMA CELL LINES: DEVELOPING THE OPTIMAL

RADIOTHERAPY PROTOCOL ___________________________________________________________ 93

SMICH, J., SHAW, K., POIRIER, V., FLECK, A. AND MUTSAERS, A.

COMBINED VESSEL NORMALIZATION AND ONCOLYTIC VIRUS THERAPY IN THE TREATMENT OF

ADVANCED STAGE OVARIAN CANCER ___________________________________________________ 94

MATUSZEWSKA, K., SANTRY, L., PETRIK, J., BRIDLE, B. AND WOOTTON, S.

ACKNOWLEDGMENTS _______________________________________________________________ 95

1

Evaluation of laparoscopic skills in veterinary students

Kilkenny, J1., Singh, A1., Kerr, C1., Khosa, D2., and Fransson, B3.

1Department of Clinical Studies, OVC, 2Department of Population Medicine, OVC,3Veterinary Clinical Sciences, WSU

Objective - To compare FLS simulator-assessed laparoscopic skills in veterinary students with

level of training, open surgical experience, and non-surgical experiences.

Design – Single-institution, prospective cohort study.

Sample Population –145 veterinary students

Procedures – Veterinary students performed a peg transfer task, pattern cut task, and ligature

loop placement task on the FLS simulator. Self-reported open surgical experience and non-surgical

experiences were collected by means of a questionnaire. A Kruskal-Wallis test was used to detect

differences in laparoscopic skills between veterinary years, and the relationship between open

surgical skills and laparoscopic skills were assessed with a Spearman’s rank order correlation

coefficient (rs). A general linear model, fixed effects ANOVA was used to investigate predictors

of laparoscopic skills in a simulator.

Results – Individual (p>0.05) and combined (p=0.75) laparoscopic skills scores were not

significantly different between veterinary years, and laparoscopic skills were not significantly

(p=0.19, rs=-0.11) associated with open surgical experience. Craft experience was a significant

predictor of laparoscopic skills on MISTELS task 2 (p<0.05) and total scores (p=0.008). MISTELS

scores on task 3 decreased significantly (p=0.034) with every unit increase in open surgical

experience.

Conclusions and Clinical Relevance - Open surgical experience and prior VG experience did

not enhance laparoscopic skills in this cohort of veterinary students. These results suggest that

proficiency with laparoscopy in veterinary students may require laparoscopic-specific skills

training.

2

Developmental ethanol exposure disrupts attention performance and

prefrontal neuron function in male mice

Louth, E., Bignell, W., Taylor, C. and Bailey, C.

Department of Biomedical Sciences

Developmental exposure to ethanol can produce a variety of teratogenic effects known

collectively in humans as Fetal Alcohol Spectrum Disorder (FASD). Although deficits in attention

rank among the most common and persistent neurobehavioural components of FASD, mechanisms

underlying this outcome are not known. Our objective was to determine effects of developmental

ethanol exposure on attention behaviour and prefrontal neuron function in adult male C57BL/6

mice. Mice were exposed to either ethanol (treatment) or sucrose (control) via oral gavage during

the mouse equivalent of the human second and third trimesters. We first measured attention

performance using the five-choice serial reaction time test. In these experiments, adult mice from

the developmental ethanol treatment group performed with lower accuracy during initial training

sessions and then exhibited a greater rate of omissions under challenging training conditions.

Whole-cell electrophysiological recording of pyramidal neurons located within layer VI of the

medial prefrontal cortex of these same mice found decreased intrinsic excitability and increased

responses to excitatory neurotransmission in mice from the developmental ethanol treatment

group. Comparison of behaviour and neuron physiology identified significant correlations between

attention performance and nicotinic acetylcholine receptor function in layer VI neurons of control

mice, which were not present for mice that had been exposed to ethanol during development. These

findings suggest that the role of prefrontal nicotinic signaling to support attention behaviour may

be disrupted following developmental ethanol exposure.

3

Distribution of ticks and the risk of Lyme disease and other tick-borne

pathogens of public health significance in Ontario, Canada

Clow, K.M., Ogden, N. H., Lindsay, L. R., Michel, P., Pearl, D., and Jardine, C. M.

Department of Pathobiology

Over the past two decades the northward spread of Ixodes scapularis across Ontario, Canada

has accelerated and the risk of Lyme disease has increased. Active surveillance is a recognized

and effective method for detecting reproducing populations of I. scapularis. In this study, we

conducted field sampling consistent with an active surveillance approach from May to October

2014 at 104 sites in central, eastern and southern Ontario to determine the current distribution of

I. scapularis and other tick species, and enhance our understanding of the geographic risk

associated with Borrelia burgdorferi and other tick-borne pathogens of public health significance

in this region. Ixodes scapularis was present at 20 of the 104 sites visited. Individuals of the tick

species Dermacentor variabilis, Haemaphysalis leporispalustris and Ixodes dentatus were also

collected. Ixodes scapularis were positive by polymerase chain reaction (PCR) for B. burgdorferi

at 5 sites. These sites formed a significant spatial cluster in eastern Ontario. No ticks were PCR-

positive for Borrelia miyamotoi, Anaplasma phagocytophilum and Babesia microti. This study

provides an up-to-date picture of the distribution of Ixodes scapularis and other ticks species, and

the risk of B. burgdorferi and other pathogens of public health significance in central, eastern and

southern Ontario. This information may allow for more effective surveillance efforts and public

health interventions for Lyme disease and other tick-borne diseases in this region.

4

Epidemiology of salmonella in raccoons and their environment on swine

farms and conservation areas

Bondo, K. J., Pearl, D. L., Janecko, N., Boerlin, P., Reid-Smith, R. J., Parmley, J., and Jardine, C. M.


Salmonella has been detected in the feces of many wildlife species, including raccoons

(Procyon lotor); however, little is known about the epidemiology of Salmonella in raccoons living

in different habitat types. Our objective was to investigate demographic, temporal, and climatic

factors associated with carriage of Salmonella and associated antimicrobial resistance in raccoons

and their environment on swine farms and conservation areas. Using a repeated cross-sectional

study design, we collected fecal samples from raccoons and environmental samples (soil, manure

pits, dumpsters) on 5 swine farms and 5 conservation areas once every five weeks from May to

November, 2011-2013. Salmonella was detected in 6% (279/1093) of raccoon fecal samples, 6%

(88/1609) of soil samples, 30% (21/69) of manure pit samples, and 23% (7/31) of dumpster

samples. We detected Salmonella with reduced susceptibility to antimicrobials in 5% (14/279) of

raccoon fecal, 8% (7/89) of soil, 10% (2/21) of manure pit, and 0/7 dumpster samples. Using multi-

level multivariable logistic regression analyses, we found location type was not a significant (P >

0.05) explanatory variable for Salmonella occurrence in raccoon fecal, soil, or manure pit samples.

Salmonella in raccoon fecal and soil samples were associated with rainfall, sex, season,

temperature and year with various interaction effects among these variables. We detected a variety

of Salmonella serovars that infect humans and livestock in the feces of raccoons indicating that

raccoons living near humans, regardless of location type, in southwestern Ontario may have a role

in the epidemiology of Salmonella and associated antimicrobial resistance.

5

Augmenting surveillance to minimize the burden of norovirus-like illness

in Ontario: Using TeleHealth Ontario data to detect the onset of

community activity

Hughes, S. L., and Papadopoulos, A.

Department of Population Medicine

Norovirus, commonly referred to as the winter vomiting disease, is the most common cause

of gastroenteritis worldwide. In Canada, the total number of cases reported per year is second only

to the common cold. The disease is highly infectious, requires a low infectious dose, and is well-

known to cause large outbreaks in closely confined populations. While deaths are rare,

hospitalization and long-term sequelae are more likely to occur in at-risk populations, such as the

elderly or immunocompromised. Action to reduce the number of norovirus infections per year is

required due to its health and economic burden. It is estimated that norovirus infections cost the

United States 2.5 billion CAD and the United Kingdom close to 200 million CAD per year in

health care costs alone. While laboratory surveillance is practiced in Ontario to detect norovirus

outbreaks, early detection remains a challenge. This project aims to utilize syndromic surveillance

using TeleHealth Ontario data in order to develop an early warning system mitigating the impact

of norovirus outbreaks.

6

Practices for disbudding and dehorning of dairy calves by veterinarians

and dairy producers in Ontario, Canada

Winder, C., Leblanc, S., Haley, D., Lissemore, K., Godkin, K. and Duffield, T.


Disbudding and dehorning (DD) dairy calves are common procedures performed on-farm.

Appropriate DD practices are important for both the welfare of the calf and public perception of

the dairy industry. The objective of this study was to build on a 2004 survey of Ontario bovine

veterinarians and dairy producers to determine if DD protocols had changed over the past decade,

and to examine factors associated with the adoption of pain-control. An online survey (Qualtrics,

2005. Provo, UT), with a telephone option, was conducted in the fall of 2014. Members of the

Ontario Association of Bovine Practitioners, and a subset of licensed producers of the Dairy

Farmers of Ontario were invited to participate. Veterinary clinics reported performed DD for a

mean of 29% (SD=20) of their dairy clients. Seventy-two percent of veterinarians reported

changing DD practices since 2004. Changes included increased us of: an NSAID (60%), sedation

(34%), and local anesthetic (29%). Sixty-three percent of producers reported changing DD

practices in the past 10 years. Changes included the addition of local anesthetic (60%), sedation

(23%) or an NSAID (17%), and performing DD at a younger age (30%). The most common

influence cited for changes was the herd veterinarian (73%). The use of local anesthetic in this

survey was far higher than previously reported (62% vs. 22%). NSAID use by veterinarians was

also higher than previously reported (59% vs. 1.5%). A strong relationship between producer and

veterinarian was associated with adoption of pain-control. Identifying factors associated with best

practices may help veterinarians target appropriate educational opportunities for their dairy clients.

7

Perioperative outcome and complications following laparoscopic

cholecystectomy in dogs: 20 cases (2008-2015)

Scott, J.1, Singh, A.1, Mayhew, P. D.2, Case, J. B.3, Runge, J. J.4, and Gatineau, M.5

1Department of Clinical Studies, Ontario Veterinary College, 2Department of Surgical and Radiological Sciences, University of California -Davis, 3Department of Small Animal

Clinical Sciences, College of Veterinary Medicine, University of Florida, 4Department of Clinical Studies, Matthew J. Ryan Veterinary Hospital at the University of

Pennsylvania,5Centre Vétérinaire DMV.

Objective - To report the perioperative outcome and complications associated with dogs

undergoing elective laparoscopic cholecystectomy (LC).

Design - Retrospective study.

Animals – 20 client-owned dogs.

Procedures – Medical records of dogs that underwent LC were reviewed for signalment, history

and clinical, ultrasonographic, and surgical findings. In all cases laparoscopic exploratory was

performed using a multiport technique. Cystic duct dissection was performed with articulating

dissectors or right angled forceps. Cystic duct ligation was performed using surgical clips and

suture. Gall bladder dissection was performed with blunt dissection, vessel sealant device or

harmonic scalpel.

Results –6 dogs (30%) required conversion from LC to an open laparotomy (OC) due to inability

to ligate the cystic duct (n=3), evidence of gallbladder rupture (n=1), leakage from the cystic duct

during dissection (n=1) and cardiac arrest (n=1). 1 dog developed bile peritonitis 6 days post

operatively following LC. A four-port technique was used in 11 dogs, three-port in 7 dogs and

two-port 2 dogs. 7 dogs had placement of a SILS port. Cystic duct dissection was performed using

articulating dissector n=10 dogs, right angle forceps n=5 dogs. Cystic duct ligation was performed

using surgical clips n= 9 dogs, suture n= 6 dogs. All dogs were discharged from hospital and had

resolution of clinical signs.

Conclusions and clinical relevance – Using careful case selection, LC can be performed

successfully for uncomplicated gall bladder disease in dogs. The surgeon should be familiar with

a variety of methods for cystic duct ligation during LC to improve success rates.

8

A systematic review and meta-analysis of on-farm methods for the

detection of hyperketonemia in dairy cattle

Tatone, E.1*, Gordon, J. 1, Hubbs, J.1, DeVries, T.2, LeBlanc, S.1, and Duffield, T.1

1Department of Population Medicine, 2Department of Animal Biosciences

Several methods have been validated for the detection of hyperketonemia (serum beta-

hydroxybutyrate (BHB) > 1.2 mmol/L) in dairy cattle on-farm, however the reported sensitivity

and specificity of each method varies and no single study has compared them all. The objective of

this work was to perform a systematic review and meta-analysis to determine the method with the

highest combined sensitivity and specificity, and the optimal threshold for each method. A

comprehensive literature search resulted in 5,196 references. After removing duplicates and

performing relevance screening, 23 studies were included for the qualitative assessment and 17 for

the meta-analysis. The diagnostic accuracy was compared using the hierarchical summary receiver

operator characteristic (HSROC) method. Subgroup analysis was conducted for each index test to

examine the accuracy at different thresholds. The Precision Xtra® device had the highest summary

sensitivity (95%) and specificity (99%). The threshold employed (1.2-1.4 mmol/L) did not impact

the diagnostic accuracy of the test. The Ketostix® and KetoTest® strips were not different in overall

diagnostic accuracy and both methods had the highest summary sensitivity and specificity when

the trace and weak positive thresholds were used The Precision Xtra® had the highest test accuracy

of the on-farm methods included in the meta-analysis. The large variation in study results

reinforces the need for diagnostic test accuracy meta-analysis.

9

Characterization of MAC-T cells as a new model for the study of TGFβ

function in bovine mammary gland involution

Mitz, C., and Viloria-Petit, A.

Department of Biomedical Science

The bovine mammary gland undergoes episodes of remodeling, which involve transient

disruption of apical-basal polarity that parallels periods of massive cell death (apoptosis). This is

prominent during involution in the dry period, the time between halting of milk removal and the

subsequent calving. Transforming growth factor-β (TGFβ) is a conserved cytokine with

demonstrated roles in mammary gland involution and apoptosis. It is well documented that TGFβ

signaling can disrupt apical-basal polarity through canonical (Smad) and non-canonical signaling,

including activation of the Par6 pathway. Additionally, our group previously demonstrated that

the Par6 pathway mediates TGFβ-induced apoptosis in mammary cells. Based on this, we

-induced apoptosis in the involuting mammary gland is linked to its

capacity to disrupt apical-basal polarity. This study examined the potential use of the immortalized

bovine mammary epithelial cell line MAC-T as a model to address this hypothesis. MAC-T cells

were cultured either as monolayers or as three-dimensional (3D) structures on reconstituted

basement membrane (rBM). Monolayers were treated with 5 ng/mL of TGFβ1 for 24 hours, and

the expression of polarity markers Zonula Occludens-1 and E-cadherin, as well as

activation/expression of TGFβ signaling mediators, Smad2 and Par6, was examined by

immunoblotting. Patterns of Smad2 activation and expression of polarity markers in response to

-like structures on rBM indicate suitability of MAC-T cells

for the intended studies. Future studies will aim to identify a polarity-associated cell survival

signaling axis involving TGFβ and Par6.

10

Biomechanical and radiological comparison between transarticular screw

fixation and 2 multi-implant constructs for ventral atlantoaxial

stabilization in the dog

Leblond, G.1, Gaitero, L.1, Moens, N. M. M.1, Linden, A. Z.1, James, F. M. K1, Monteith, G.1, and

Runciman, R. J.2

1Department of Clinical Studies, 2School of Engineering

Numerous techniques have been described to stabilize canine atlantoaxial joint, yet there is

currently little data comparing these methods. The objective of this study was to compare the

biomechanical properties, and technical outcome of 3 ventral atlantoaxial stabilization techniques

in canine cadavers. 21 Beagle dog cadavers were randomly assigned 1 of 3 stabilization techniques

including transarticular screw fixation (TSF) and 2 multi-implant constructs (MI5 or MI6)

respectively composed of 5 or 6 screws embedded in polymethyl-methacrylate cement. All screws

were placed based on another study defining ideal implant placement using a new drill guide

prototype designed to improve accuracy of implant placement. C1-C2 spinal segments were then

extracted and computed tomography images were obtained to assess for anatomical reduction and

safety of implant placement. Ventral bending moment at failure and stiffness of the constructs

were then determined and compared between groups. Ventral stabilization was achieved

successfully in all cadavers. No major technical issues were encountered using the drill guide

prototype. Significant vertebral canal violation was identified in 4/91 screws (4.4%). Intervertebral

apposition was significantly better in the TSF group compared to MI5 and MI6 (p=0.018). Bending

moment at failure was lower in TSF group (3.66±1.23N.m) compared to MI5 (8.51±1.96N.m;

p=0.0004) and MI6 (7.73±2.31N.m; p=0.0021). Mean stiffness measured in the 0-5° region was

similar in all groups. TSF provided better intervertebral apposition but was significantly less

resistant to ventral flexion compared to 2 multi-implant fixation techniques in a Beagle cadaver

model.

11

Comparative genomic analysis of two netF-positive Clostridium

perfringens isolates associated with foal necrotizing enteritis and canine

haemorrhagic gastroenteritis.

Mehdizadeh Gohari, I.1, Kropinski, A.1, Weese, S.1, Parreira, V.1, Whitehead, A.2, Boerlin, P.1, and

Prescott, J.1

1Department of Pathobiology, University of Guelph, 2Department of Veterinary Clinical and Diagnostic Sciences, University of Calgary

The recent discovery of a novel beta-pore-forming toxin, NetF, which is strongly

associated with canine and foal necrotizing enteritis should improve our understanding of the role

of type A Clostridium perfringens strains in disease in these animals. The current study presents

the complete genome sequence of two netF-positive strains, JP55 and JP838, which were

recovered from a case of foal and canine necrotizing enteritis, respectively. Genome sequencing

was done using Single Molecule, Real-Time (SMRT) technology-PacBio and Illumina Hiseq2000.

The JP55 and JP838 genomes include a single 3.34 Mb and 3.53 Mb chromosome, respectively,

and both genomes include also five circular plasmids. Comparison of these two C. perfringens

chromosomes with three available reference chromosome sequences identified 48 (~ 247 kb) and

81(~ 430 kb) regions unique to JP55 and JP838, respectively. Some of these divergent genomic

regions in both chromosomes are phage- and plasmid-related segments. Sixteen of these unique

chromosomal regions (~69 kb) were shared between the two isolates. In addition, plasmid

annotation revealed that three plasmids were shared by the two newly sequenced genomes,

including a NetF/NetE toxins-encoding plasmid, a CPE/CPB2 toxins-encoding plasmid and a

putative bacteriocin-encoding plasmid. Comparative analysis showed that the beta-pore forming

toxins genes, netE, netF and netG, were located in unique pathogenicity loci on tcp-conjugative

plasmids. These results provide significant insight into the basis of canine and foal necrotizing

enteritis and are the first to demonstrate that netF resides on a large and unique plasmid-encoded

locus.

12

The primary benefits of angiotensin-converting enzyme inhibition on

cardiac remodeling occur during sleep time in murine myocardial

ischemia-reperfusion (heart attack)

Tsimakouridze, E., Alibhai, F., Reitz, C., and Martino, T.

Centre for Cardiovascular Investigations, Department of Biomedical Sciences, University of Guelph

Introduction: Many commonly prescribed drugs target the products of circadian genes, supporting

the new drug treatment strategy that chronotherapy (time-of-day treatment) can benefit patients

after myocardial infarction (MI, heart attack).

Hypothesis: Angiotensin converting enzyme inhibitors (ACEi), commonly prescribed to patients

post-MI, reduce adverse cardiac remodeling when taken at sleep-time instead of conventional

wake-time.

Methods and Results: To demonstrate the benefit of time-of-day ACEi we used the murine

myocardial ischemia-reperfusion (I/R) model and administered captopril at sleep vs wake-time.

We show that 1) sleep-time captopril administration better maintains heart weight:body weight

post-IR as compared to wake-time captopril. 2) Moreover, sleep vs wake-time captopril improves

heart structure and function, as demonstrated by echocardiography with decreased left ventricular

(LV) internal dimensions at diastole (4.51±0.04mm vs 4.86±0.13mm) and increased % ejection

fraction (62.21±1.10 vs 55.70±1.72). 3) Sleep vs wake-time captopril also better maintains LV

end-systolic volume (24.88±1.05ul vs 39.07±4.34ul) as shown by hemodynamic measurements.

4) Lastly, sleep vs wake-time captopril benefits scar morphometry as histological analysis showed

increased infarct thickness (2.37±0.19cm vs 1.56±0.13cm). To understand the underlying

mechanism of why sleep-time ACEi therapy is more beneficial than wake-time, we will use

microarrays and bioinformatics analyses.

Conclusion: We show that sleep-time ACEi improves cardiac structure, function, and outcome

post-I/R compared to wake-time ACEi and placebo. This novel strategy can benefit human and

animals with heart disease.

13

The effect of immunological status, in-vitro treatment and culture time on

expression of eleven candidate reference genes in bovine blood

mononuclear cells

Emam, M., Thompson-Crispi, K., and Mallard, B.


Feasibility of RNA quantification by real time RT-PCR has led to enormous utilization of this

method. However, real time PCR results need to be normalized due to the high sensitivity of the

method and also to eliminate technical variation. Normalization against a reference gene that is

constitutively transcribed and has minimum variation among samples is the ideal method.

Nevertheless, many studies have shown that there is no general reference gene with ideal

characteristics and candidate reference genes should be tested before being used as a “normalizer”

in each study. The current study investigated the effects of previous exposure of the host to

experimental test antigens and culturing time on the expression of 11 candidate genes when bovine

blood mononuclear cells (BMC) were cultured and treated in-vitro by two different antigens and

a mitogen. Our findings showed that the expression of the most common reference genes, β-actin

and GAPDH, are significantly decreased at 24 hours after culturing BMCs, even without any

treatment. After treatment of cultured BMCs with C. albicans whole yeast extract and Hen Egg

White Lysozyme (HEWL), only the expression of C-terminal binding protein 1 (CTBP1) and

RAD50 among all tested genes were consistent, respectively. In addition, expressions of CTBP1

and RAD50 were independent from previous exposure of the host to the antigen. The results of

this study demonstrated inconsistent expression of commonly used reference genes in untreated

cultured BMCs over time. As this condition applies to negative controls in real time RT-PCR study

designs, normalization against these genes can largely deceive the outcome, especially in kinetic

studies.

14

Characterization of the equine contact network at a single equestrian

show

Spence, K.L., Goh, B., O’Sullivan, T.L., Poljak, Z., and Greer, A.L.


Identifying the risk of infectious disease introduction and spread in the Ontario equine

population depends on understanding how horses contact one another as they travel. However, the

lack of available information on equine contact patterns in Ontario limits the ability to assess the

risk of a disease outbreak in the population. The objective of this study was to characterize the

network of potential contacts associated with a single equestrian show to determine how network

structure may affect equine influenza (EI) transmission. Horse show participants completed a

questionnaire asking about their horse’s “home” facility, travel patterns, and biosecurity practices.

Social network analysis was used to characterize the network, which included horses attending the

show and horses stabled at each of their home facilities (secondary contacts). Responses were

received for 80% (55/69) of horses attending the show. The two-mode undirected network

consisted of 820 nodes (41 locations and 779 horses). Eight percent of nodes in the network

represented horses attending the show, while 87% of nodes represented secondary contacts. The

median (range) degree of a horse in the network was 33 (1-105). The horse with the highest

centrality scores had contact with horses in three locations, indicating that it was the most

important node for potential disease spread in the network. An agent-based disease transmission

model is currently being developed to simulate the impact of an inadvertent introduction of EI to

an equestrian show. Examining the connectivity between horses attending an equestrian event is a

fundamental step in the identification of effective disease prevention strategies.

15

Reproducibility, stability and reference ranges of thrombin generation

using the calibrated automated thrombogram in a population of healthy

dogs.

Cuq, B.1, Blois, S.1, Wood, D.2, Abrams-Ogg, A.1 and Bédard, C.3

1Departments of Clinical Studies 2Department of Pathobiology,3Department of Pathology and Microbiology, University of Montreal

A current challenge in human and veterinary medicine is the early identification and treatment

of patients prone to thromboembolisms. Thrombin plays a central role in hemostasis and

thrombosis, and is involved in all in vivo hemostatic pathways. Calibrated automated

thrombogram (CAT) is a thrombin generation (TG) assay, and is a valid physiological test for

hemostasis. CAT results can be influenced in humans by several pre-analytical variables,

especially blood collection method. Objectives were to establish reference intervals for CAT

thrombin generation, and validate this method including assessment of pre-analytical and

biological variability in 41 healthy dogs. Two sampling methods were performed for each dogs:

direct venipuncture using a 21ga needle (jugular vein) and venipuncture using a 21ga butterfly

needle (saphenous vein). CAT assay consisted of the use of a fluorogenic thrombin substrate. TG

was initiated with the addition of 5pM of tissue factor. Storage stability was assessed at 6 time

points on 16 samples. 9 dogs had blood collected weekly to assess the intra-individual variability

of CAT at 4 time points for both methods. No significant difference between the different storage

lengths: 24h at 4oC; 24h, 1 week, 1 month or 2 months at -80oC compared to baseline could be

found. No significant difference was found between week 1, 2, 3 and 4. Interclass correlation

coefficients showed a poor repeatability for butterfly-obtained samples compared to direct

venipuncture. Coefficient of concordance showed poor agreement between jugular and saphenous

sample. CAT using samples obtained by direct venipuncture is stable and repeatable in healthy

dogs. Butterfly catheter assisted venipuncture is not a repeatable method and there was no

agreement with the direct venipuncture.

16

Structural and Functional Integrity of Microwave Ablation Antennas with

Repeated Use and Re-sterilization

Finck, C., Zur Linden, A., Singh, A., Foster, R. and Nykamp, S.

Department of Clinical Studies

Microwave ablation (MWA) is performed in humans as a minimally invasive treatment of

various cancers, and has been shown to reduce overall cost, morbidity and mortality compared to

open surgical procedures. A single use MWA antenna is ~$3200 CAD, which considerably limits

its use in veterinary patients. The objective of our study was to determine the structural and

functional integrity of MWA antennas with repeated use and re-sterilization. 17 saline cooled

MWA antennas were divided into three groups of five antennas and one group of two controls.

For each cycle, one, two and three ablations were performed in bovine cadaver livers for each

group respectively, except the controls. Antennas were cleaned then sterilized in hydrogen

peroxide plasma. Aerobic and anaerobic bacterial cultures were performed and the antennas

assessed for damage with stereo light microscopy. Four reprocessing cycles were completed. No

significant changes in the size of the ablation zones were noted (p=0.064). Progressive tearing of

the silicone coating the antennas was observed over the cycles with a negative influence of the

number of cycles on silicone tearing (p=1.12x10^-9). No bacterial growth was present. 13/15

MWA antennas remained functional for up to 4 reprocessing cycles when used at the maximum

recommended settings. Clinical significance of the tearing of the silicone coating remains to be

determined. The structure and function of the antennas remained intact during repeated use and

reprocessing, suggesting this practice could be performed in a clinical and research setting at the

clinicians discretion.

17

Combining Oncolytic Viruses with Epigenetic Modifiers as a Novel Therapy

for Leukemia

Strachan-Whaley M, Au Yeung A, Kim J, Bienzle D, Mutsaers A, Wootton S, Bridle B


Acute lymphoblastic leukemias (ALL) are a group of blood cancers that are derived from B-

cells (B-ALL), T cells (T-ALL), and rarely natural killer (NK) cells. There are many different

types of acute myeloid leukemias (AML). Companion animals, including canines and felines, can

also develop acute leukemias, but these diseases are poorly characterized and the prognoses remain

dismal. This study seeks a therapy for canine and feline acute leukemias. Oncolytic viruses (OV)

are viruses that specifically target tumour cells. This therapy can be very effective in treating solid

tumours, but performs poorly in leukemias where the cells are widely dispersed among normal

tissues with anti-viral defenses that can quench the infection. It has been established that epigenetic

modifiers can modulate the replication of OVs in solid tumours to enhance efficacy. In a reversal

of this paradigm, we propose that only limited infection with an OV will induce a robust anti-viral

response that can sensitize leukemic cells to the effects of epigenetic modifiers. The first aim of

this project is to assess the therapeutic efficacy of OVs and epigenetic modifiers as monotherapies

and in combination in vitro Initial studies were carried out in a mouse B-ALL cell line, and optimal

combinations have been identified. These combinations are currently being further tested in canine

and feline leukemia cell lines. Optimal combinations are also currently being tested in vivo using

a mouse model of B-ALL, and will be tested ex vivo in leukemic canine and feline blood samples.

The ultimate goal is to establish a therapy that could potentially proceed into a canine or feline

clinical trial.

18

Factors affecting the composition of the rabbit fecal microbiota

Kylie, J., Weese, J. S. and Turner, P.V.


The health and disease status of all mammals is significantly influenced by the gastrointestinal

(GI) microbiota. In addition to optimizing nutrient absorption from food, it also aids in developing

the GI immune system, protecting the host against invasion of pathogenic organisms. By studying

the GI microbiota of rabbits, we can begin to identify how changes within its composition can

influence the health of rabbits. The aim of the current study was to characterize the fecal microbiota

of rabbits and to identify how factors such as age, season, source, and feed were likely to influence

the composition. Pooled fecal samples were collected from rabbits from commercial farms

(n=100), laboratories (n=14), local shelters (n=15), and from pet rabbits (n=60). Following

bacterial DNA extraction, culture-independent 16s rRNA gene sequencing of the V4 region was

conducted on all samples to identify the components of the fecal microbiota as a reflection of the

overall GI microbiota. The composition of the fecal microbiota was consistent across all sources

with the predominant phylum being Firmicutes (65-73% of the total composition). Proteobacteria,

a phylum composed predominantly of pathogenic bacteria species, was significantly more

predominant in commercial rabbits than rabbits from other sources (14% vs 4-8%) and in summer

versus winter months (19% vs 9%). The proportion of Verrucomicrobia, a phylum associated with

fibrolysis, was significantly higher in rabbits routinely fed hay and vegetables. These results raise

a number of interesting questions regarding the impact that the rabbit fecal microbiota and various

factors that influence it have on predisposition to disease.

19

Post-exercise cardiac troponin i release and clearance in normal

standardbred racehorses

Rossi, T. M.1, Pearl, D. L.1, Pyle, W. G.2, Maxie, M. G.3, Kavsak, P. A.4, and Physick-Sheard, P. W.1

1Department of Population Medicine, Ontario Veterinary College, 2Department of Biomedical Sciences, Ontario Veterinary College, 3Animal Health Laboratory, Laboratory

Services Division, University of Guelph, 4Department of Pathology and Molecular Medicine, McMaster University

Reasons for performing study: There are currently no studies detailing cardiac troponin I (cTnI)

release and clearance in normal horses post-exercise using an analytically validated assay. These

data are essential for selecting appropriate sampling times in equine athletes with suspected

myocardial damage.

Objective: To plot the magnitude and time course of cTnI release and clearance, using a validated

cTnI assay, after maximal effort.

Methods: Five clinically normal Standardbred racehorses in full race training were included in the

study. Physical examinations were performed on subjects and blood samples were taken via

jugular venipuncture pre-exercise. Horses were exercised in harness at race intensity in groups on

a training track. A second blood sample was taken immediately post-exercise and an intravenous

catheter was then placed in a jugular vein. Hourly blood samples were taken for 24 hours. All

samples were collected in red top serum Vacutainer® tubes. Serum samples were stored at -80°C

until analysis. All samples were analyzed using the Abbott ARCHITECT STAT High Sensitivity

Troponin-I assay.

Results: Mean resting cTnI level was 1.33 ± 0.6 ng/L (range, 0.82 - 2.33 ng/L). All horses exhibited

an increase in cTnI level after exercise with peak elevation occurring 2 - 6 hours post-exercise

(mean, 4.6 ± 1.7 hours). Mean peak increase in cTnI level was 11.96 ± 9.41 ng/L (range, 1.72-

23.76 ng/L). All horses returned to baseline levels within 24 hours.

Conclusions: All horses experienced an increase in cTnI post-exercise, with the peak occurring 2-

6 hours post-exercise. Further studies are needed to determine the significance of these increases.

20

Understanding the mechanism behind the tissue specificity and

oncogenesis of JSRV and ENTV in sheep

Rosales Gerpe, M. R.1, Walsh, S. R.2, de Jong, J.1, and Wootton, S. K.1

1Department of Pathobiology, 2McMaster University

Jaagsiekte Sheep Retrovirus (JSRV) and Enzootic Nasal Tumour Virus (ENTV) are the

causative agents of two economically important cancers that affect sheep and goats. Their

Envelope (Env) glycoprotein is a potent oncogene that can rapidly transform cells in vitro and in

vivo through an as yet unidentified mechanism. Nevertheless, many of the signaling pathways

activated in lung and other types of cancer are also turned on in Env-transformed cells. Therefore,

studying these viruses is of great importance not only because they are animal pathogens, but

because their Env oncoprotein provides an excellent tool for probing mechanisms involved in

carcinogenesis. Though they induce tumours in different parts of the sheep respiratory tract, they

share high nucleotide and amino acid identity. To understand what drives tissue specificity, I have

generated JSRV-ENTV hybrid viruses in which regions of the JSRV genome have been replaced

with the corresponding ENTV sequences. These chimeras will be evaluated ex vivo and in vivo for

their ability to transform lung or nasal epithelium. To understand the mechanism involved in Env-

mediated transformation, I will perform co-IP and PLA experiments with JSRV Env and a list of

candidate binding partners previously identified in a yeast-2-hybrid screen. The biological function

of these interactions will also be analyzed in vitro and in vivo. Overall, this work will expand our

knowledge of betaretroviral pathogenesis and cellular transformation, and may lead to practical

applications for gene therapy.

21

Monitoring differences in estrous cycle length in summer and winter

seasons by measuring progesterone in water buffaloes

Dharap, A.1, Gill, I.1, Moradi, F.1, Ashkar, F. A.1, Robertson, L.1, West, C.2, Littkemann, M.2, Smith,

L.2, Antenos, M.1, and King, W. A.1

1Department of Biomedical Sciences, Ontario Veterinary College, 2Ontario Water Buffalo Company

Background: Water buffaloes produce good quality milk, which is a growing market in Canada.

Although they are capable of breeding throughout the year, increased conception rates have been

observed in winter compared to summer. Reproductive endocrine hormone profile in milk, as a

non-invasive method, has been established. Having been introduced to the Canadian environment,

water buffalo reproduction will be seasonally affected and analysis of milk progesterone profiles

can shed light on seasonality. Present study sets out to determine ovarian follicular dynamics by

monitoring the progesterone peak and its stability during summer (long days) vs. winter (short

days) to predict pregnancy and status of estrous cycle.

Methods: Milk was collected from randomly selected buffaloes during breeding season (n=3)

(November 25/2014 – January 2/2014) and non-breeding seasons (n=3) (May 21/2014 – June

21/2014). Progesterone was measured every day in summer and every other day in winter via an

ELISA assay.

Results: Summer months show abrupt rise and fall in progesterone with short elevated intervals

and remained low throughout the reproductive cycle. Progesterone peak elevation lasted 2 days.

Winter months showed an increase in progesterone with maintenance of peak over 10 days. Estrus

cycle length in winter was consistently 20 days among all buffaloes.

Conclusions: Preliminary results indicate that in non-breeding seasons, irregularity in

progesterone levels is seen with an instable peak. In breeding seasons, high levels of progesterone

were maintained over several days indicating active corpus luteum. Estrus cycle length was longer

in winter with a stable elevated progesterone peak.

22

Characterization of the TGFβ-TAZ signalling axis in canine osteosarcoma

cell lines

Andrade, A. D.1, Neogi, P.1, Mutsaers, T.1, Wood, G.2, and Viloria-Petit, A.1

1Department of Biomedical Sciences, 2Department of Pathobiology

Osteosarcoma (OSA) is the most common bone tumor in dogs, where it commonly

metastasizes to the lungs. Metastatic canine OSA is resistant to chemotherapy and is responsible

for patient mortality. OSA metastasis and chemoresistance mechanisms are generally unknown.

Transforming growth factor-β (TGFβ) is a highly conserved cytokine with roles in bone

development, bone disorders, cancer metastasis and chemoresistance. The action of TGFβ is

carried out through Smad transcription factors, which rely on TAZ to alter gene expression. TAZ

is a highly conserved transcriptional coactivator and an integral part of the Hippo pathway

controlling organ size, proliferation and differentiation, as well as bone mass and

osteoblastogenesis. TAZ was shown to mediate self-renewal capacity and cancer metastasis. We

hypothesized that the TGFβ-TAZ signalling axis mediates OSA progression and chemoresistance.

generated by our group. Next, we treated the cells for 2, 24 and 48 hours with 0, 0.5 and 5 ng/mL

TGFβ1, and determined the pattern of Smad2 activation and TAZ expression using

immunoblotting. 24-hour treated cells were also examined for their colony-forming ability after

doxorubicin treatment. All cell lines expressed TGFβ receptors and showed a robust TGFβ1

response, displaying dose-dependent Smad activation that varied with time and cell line. 24-hour

TGFβ1 treatment caused a dose-dependent TAZ increase in the majority of cell lines, and impacted

their ability to form colonies after doxorubicin treatment. Futures studies will investigate the

TGFβ-TAZ signalling axis in chemotherapy resistance.

23

Health-seeking behaviour in response to acute gastrointestinal illness

among the ugandan batwa

Weber, L.J. and Indigenous Health Adaptation to Climate Change (IHACC) Research Team


The Batwa, an Indigenous people who inhabit the southwest highlands of Uganda, were

recently forcibly evicted from their traditional, forested lands. Due to national conservation

projects, approximately 6,700 Batwa individuals were forced to transition from their former

hunter-gatherer lifestyle to an agricultural livelihood. This displacement eliminated access to

traditional food, medicines, and shelter, and shifted healthcare services for the Batwa. While some

individuals still rely on traditional medicines, others seek treatment at a nearby Western healthcare

facility. The purpose of this exploratory study was to examine why Batwa individuals choose one

form of healthcare over the other, or choose not to seek any healthcare, in response to Acute

Gastrointestinal Illness (AGI), which is highly prevalent in this population. From June 1 to July

30, 2014, field researchers from the IHACC project conducted 10 focus groups in 5 Batwa

settlements, and 11 key informant interviews, with healthcare workers, community health

coordinators, and clinical officers. This qualitative data was then analyzed with thematic analysis,

using a constant comparative method.

This study not only provided insight as to the Batwa’s perceptions of Western and traditional

medical care, but also created a deeper understanding of the barriers they experience to accessing

either form of care. Known as one of the world’s most vulnerable populations, the Batwa’s recent

displacement poses significant risks to their health and wellbeing. This study is intended to inform

public health interventions to reduce the burden of AGI and restore adequate healthcare,

biomedical or traditional, to the Batwa.

24

A longitudinal study of the influence of flavophospholipol on salmonella

shedding and colonization in naturally infected pigs

Nair, S., Farzan, A., O’Sullivan, T., and Friendship, R.M.


Background The heightened prevalence of multi-drug resistant Salmonella spp. in swine is

detrimental to food safety. Research suggests that flavophospholipol, a non-transferable antibiotic,

can alter the gut flora in favor of beneficial bacteria inhibiting the colonization of Salmonella in

animals.¹

Objective To investigate whether in-feed flavophospholipol can reduce the Salmonella shedding

and colonization in pigs.

Methods Forty-five 9-week-old pigs were purchased from a farm with Salmonella history and

housed at Ponsonby Animal Research Facility. Over the span of 10 weeks the pigs either received

4ppm of flavophospholipol (Flavomycin®, Huvepharma) in-feed (n=25) or a non-medicated feed

(n=20). Weekly fecal samples as well as various tissue samples were collected at slaughter and

cultured for Salmonella.

Results Salmonella was cultured from 89% (40/45) of pigs more than once; one pig tested positive

8 times. At slaughter, of 43 pigs, 6 tested positive for Salmonella in at least one tissue culture.

Salmonella was cultured from spleen, liver, lymph nodes, tonsil, and cecal content. The mean

prevalence of Salmonella shedding over 10 weeks in pigs that received flavophospholipol (27%)

was not significantly different from that in the control pigs (28%) (P>0.05).

Conclusion Naturally infected pigs subsequently given flavophospholipol at 4ppm in feed continue

to shed Salmonella and harbour Salmonella at slaughter similar to non-medicated pigs. However,

further studies are needed to investigate the preventive effect of flavophospholipol if administered

before pigs were infected with Salmonella.

25

Rheological properties of a sustained-release formulation of butorphanol

in a poloxamer 407 base and pharmacokinetics in amazon parrots

(amazona ventralis)

Laniesse, D.¹, Sanchez-Migallon Guzman², D., Smith, D.¹, Knych, H.², Mosley, C.¹, Paul-Murphy, J.²,

and Beaufrère, H.¹

¹Health Sciences Center, Ontario Veterinary College, ²Department of Veterinary Medicine and Epidemiology, School of Veterinary Medicine, University of California-Davis

Butorphanol is an opioid drug used routinely for pain management in psittacine birds.

However, the short 1-3 h period of antinociception demonstrated in Amazon parrots limits its

clinical application. Moreover, butorphanol is usually administered parenterally SQ, IM or IV as

a continuous rate infusion, due to its low oral bioavailability. Repeated handling is stressful,

multiple injections may induce pain, and IV catheter placement might not be feasible is some

situations, hence the need for a sustained-release opioid analgesic in psittacine birds. Poloxamer

407 (P407, Sigma-Aldrich) hydrogel has reverse gelation properties and functions as a sustained

release drug carrier. The rheological properties of P407 and P407-butorphanol were determined

over a range of temperatures. Butorphanol did not significantly alter the viscosity of the hydrogel,

and all formulations were in gel form at avian body temperature (39-41 °C). Sterilization of the

gel using 0.22μm filter units did not significantly alter its viscosity. Pharmacokinetic parameters

of a dosage of 15 mg/kg butorphanol tartrate in 25% P407 administered SQ were evaluated in

eight Hispaniolan Amazon parrots. Dosage and P407 concentration were based on the results of a

series of pilot studies. Blood samples were collected before injection, and at 0.08, 0.5, 1, 2, 4, 8

and 12h following drug administration. Plasma butorphanol concentrations were measured using

high-performance liquid chromatography – mass spectrometry. The results indicate that plasma

concentrations of butorphanol considered therapeutic can be reached, and that the duration of

action of P407-butorphanol may be longer than that of butorphanol tartrate.

26

T lymphocyte responses to mycobacterium avium subspecies

paratuberculosis (MAP) -derived culture filtrate proteins and membrane

vesicles during early intestinal MAP infection

El Skhawy, N., and Plattner, N.


Johne’s disease (JD) is chronic, progressive and eventually fatal granulomatous enteritis of

domestic ruminants caused by Mycobacterium avium subspecies paratuberculosis (MAP).

Infected animals progress through a lengthy subclinical infection period, during which definitive

diagnosis is virtually impossible. Importantly, progressive shedding of live MAP in the feces

occurs during subclinical infection, which represents a major source of bacteria for susceptible

herd mates and emphasizes the importance of early disease diagnosis for the control of this disease.

Host‐pathogen interactions and the development of anti‐mycobacterial T cell responses during

early intestinal MAP infection remain poorly understood, and this hinders development of novel

diagnostic testing strategies. Our hypothesis is that the early host immune responses contribute

directly to early MAP clearance from the host, and can be used to predict disease outcome.

Researchers have studied lymphocyte responses to MAP crude antigen or cell wall components;

however the role of MAP early‐secreted antigens during initiation of host immunity remains under

explored. To address this knowledge gap, we will screen T cells harvested from experimentally

infected calves against several potential early secreted antigens of MAP, including purified culture

filtrate (secreted) proteins and membrane vesicles obtained from MAP cultures. Our objective is

to correlate T lymphocyte responses to secreted MAP antigens with disease progression in the

intestine in order to understand development of early host T cell immunity and to identify antigens

with potential use in development of improved diagnostic testing strategies.

27

Development of orf virus (parapoxvirus ovis) as a multivalent viral vector

platform against Toxoplasma gondii

Ingrao, J.C., van Vloten, J.P., Shapiro, K., Barta, J.R., Menzies, P.I., Bridle, B.W., and Wootton, S.K.


Toxoplasma gondii is one of the most common causes of abortion in sheep in Canada.

Economic loses are significant and there are no effective means to prevent T. gondii in Canada.

Orf virus (ORFV) is a promising vaccine vector for infections requiring a predominantly TH1 cell-

mediated immune response. We hypothesize that reprogramming ORFV to express heterologous

antigens will stimulate a potent T-cell mediated immune response, thereby reducing tissue cyst

burden and/or abortion incidence in sheep. We aim to 1) develop ORFV which expresses T. gondii

antigens, 2) optimize T. gondii-expressing ORFV to elicit our desired immune response 3)

demonstrate efficacy of the vaccine in sheep. T. gondii antigens (SAG1, ROP2, and GRA2) will

be inserted into the VEGF loci (thereby removing this viral virulence factor). To develop a transfer

plasmid, regions flanking the VEGF gene have been sequenced, amplified, and cloned into

pBluescript. The T. gondii antigens have been developed as a gene block cassette, amplified and

cloned as a polyprotein separated by 2A cleavage sequences, and cloned into the transfer plasmid.

Insertion of T. gondii antigens will occur via homologous recombination and facilitated by the use

of the CRISPR-Cas9 system. Concurrently, the T. gondii antigen cassette has been cloned into an

Ad48 vector, which will be used for prime-boost regimens. BALB/c mice will be vaccinated in

order to evaluate safety, to characterize the immune response, and to determine ideal prime-boost

regimen. We will then perform a challenge study (M4 T. gondii strain) in pregnant sheep and

evaluate tissue cyst burden and abortion rate.

28

Harnessing immunogenic cell death to potentiate anti-cancer efficacy

during ORFV-induced oncolysis.

van Vloten, J. P., Bridle, B.W., and Wootton, S.K.


Immunogenic cell death (ICD) is a recently uncovered phenomenon in which cells committed

to death undergo a distinct cascade of events that culminate in the release of factors that alert the

immune system. This is in contrast to conventional apoptosis, which is immunologically silent.

Studies investigating the effect of anthracycline-induced cancer cell death demonstrated that

translocation of calreticulin (an endoplasmic reticulum protein) to the cell surface and extracellular

secretion of high-mobility group box 1 (HMGB1; a nuclear DNA-binding protein) drive

immunogenicity. Surface calreticulin acts as a pro-phagocytic signal, leading to increased

processing of tumor antigens by dendritic cells. Outside of the nucleus, HMGB1 functions as either

a chemokine or pro-inflammatory cytokine depending on its oxidation state. Together, calreticulin

and HMGB1 translocation enhance the immune response to tumor antigens, improving the efficacy

of anti-cancer therapies in murine models.

Oncolytic viruses preferentially infect cancer cells leading to cell death and the release of viral and

tumor antigens, acting as in situ cancer vaccines. Recent investigation has shown that combination

therapy with ICD-inducing chemotherapeutics enhances the immunogenicity of oncolytic

virotherapy, translating to improved therapeutic efficacy. We postulate that delivering the

dominant ICD signals, calreticulin to the cell surface and HMGB1 secretion, from a recombinant

oncolytic virus will mimic the improvements in combination therapy without systemic

chemotherapy. We will demonstrate this using a novel Orf virus-derived poxvirus system, in a

murine model of ovarian carcinoma.

29

Accuracy and intensity of estrus detection with activity monitoring

systems for lactating dairy cows

LeRoy, C. and LeBlanc, S.J.


Activity monitors are becoming a widely used tool for estrus detection in dairy cows. The objective

of this study is to assess the intensity and accuracy of estrus detection with automated activity

monitoring systems. Three commercial dairy farms with an activity monitoring system (Heatime,

SCR Inc, or Afi PedoPlus, Afikim) that was used for essentially all inseminations between 50 and

80 days in milk (DIM) were enrolled in this observational study. Herds were visited once weekly.

Blood samples were collected at weeks 5, 7, and 9 postpartum and from a subset of cows on the

day of insemination, to measure serum progesterone concentration. Cows were examined at week

5 for purulent vaginal discharge (PVD). Lameness and body condition were scored at week 7.

Cows were classified as anovular if all 3 blood samples from weeks 5, 7, and 9 had progesterone

< 1 ng/ml. Overall, 6.7% of cows were anovular (range of 3.6% to 8.2% among farms). Of these

cows, 39% had a lameness score of 3 or above (1 to 5 scale) and 29% had PVD. Among cows that

reached 80 DIM (n=419), 14% had not been inseminated by 80 DIM; of these, 12% were anovular,

23% were lame, 21% had PVD and 4% had a BCS of <2.5%. Among 352 blood samples taken on

the day of insemination (all services; 212 signaled by the activity system and 140 by Ovsynch),

8% and 3% had progesterone > 1 ng/ml i.e., the cows were not in estrus. In this sample, the

accuracy of detection of estrus by AAM was high, but a subset of cows would likely require

intervention for timely first AI and only a fraction of these were truly not cyclic.

30

Using an agent-based model to compare two diagnostic tests for

Staphylococcus aureus bovine mastitis

Gardner, E.1, Kelton, D.1, Hand, K.2, Poljak, Z.1, and Greer, A. L.1

1 Department of Population Medicine 2Strategic Solutions Group

Staphylococcus aureus is the most prevalent causative agent of mastitis in Canadian dairy

herds, and transmission via milking is the primary source of infection. Two milk-based diagnostic

tests are available with differing characteristics. Bacterial culture, the traditional gold standard,

and polymerase chain reaction (PCR), a recently developed test, have sensitivities of 53% and

91%, and specificities of 89% and 99%, respectively. This model aimed to explore the implications

of using different tests on the spread of S. aureus mastitis at the herd level. Using a stochastic

agent-based simulation, a population of 100 cows with homogenous mixing and a contact rate of

2 per day was modeled to reflect a typical Ontario dairy farm with twice daily milking.

Transmission probabilities were based on estimates derived from previous models. Diagnostic

testing was based on monthly screening using individual cow composite milk Somatic Cell Counts

(SCC), estimated from distributions of Ontario herds, and test positive cows were segregated from

the general herd to decrease S. aureus transmission. The choice of diagnostic test had a significant

impact on the incidence of S. aureus mastitis. The mean annual incidence was 0.42 cases per

lactation at risk with bacterial culture testing, and 0.18 cases per lactation at risk with PCR testing

(p<0.001) with 500 model iterations. Furthermore, the number of cows that were segregated was

much lower with PCR (μ=26, SD=5.2) than with culture (μ=72, SD=6.5). Thus, when combined

with effective interventions such as segregation of infectious animals, the greater sensitivity and

specificity of PCR can reduce the overall spread of S. aureus mastitis.

31

Prognostication of canine t-cell lymphoma

Deravi, N.1, Berke, O.2, Bienzle, D.1

1Departments of Pathobiology, 2Department of Population Medicine

Lymphoma is a common cancer in dogs frequently treated with chemotherapy. T-cell

lymphoma is thought to have poorer response than B-cell lymphoma. However, there are different

types of T-cell lymphoma with purportedly variable prognosis. Lymphocytes are grouped broadly

into B- or T-cell types by detection of CD79 or CD21 (B) and CD3 (T) cell markers, respectively,

using immunohistochemistry or flow cytometry. Detection of additional markers such as CD4 and

CD8 allow sub-classification of T-cells. To determine if specific T-cell lymphoma subtypes have

distinct response to chemotherapy and survival, flow cytometric data from 127 dogs with T-cell

lymphoma treated with combination chemotherapy at the Ontario Veterinary College over 12 years

were analyzed. Median overall survival time and progression free interval (PFI) were 179 and 148

days, respectively. One hundred and two dogs died from lymphoma, 25 dogs died or were

euthanized due to clinical deterioration within the first month after diagnosis, 31 survived more

than a year after start of therapy, and none died of lymphoma after surviving for two years. Absent

or low CD4 expression, high CD8 expression, high MHCII expression, and the use of single agent

or combination chemotherapy were associated with longer survival and PFI. Weight, sex, cell size,

and CD3 expression did not have a significant effect. These results suggest that specific

immunophenotypic markers correlates with survival, and certain types of T-cell lymphoma have a

very favourable prognosis. Future investigations will focus on the relationship between

immunophenotype, histopathological subtype and clinical outcome.

32

Identification of Bordetella bronchiseptica in fatal pneumonia of dogs and

cats

Bassel, L.L.¹, Abdelaziz, K.¹², Harness, M.L.¹, Clark, M.E.¹, Register, K.B.³, Caswell, J.L.¹

¹Department of Pathobiology, ²Department of Pathology, Faculty of Veterinary Medicine,

Beni-Suef University, ³USDA National Animal Disease Center

Infection with Bordetella bronchiseptica is a common cause of tracheobronchitis and upper

respiratory disease in dogs and cats, but can also lead to fatal pneumonia. Specific identification

of this pathogen is important due to risk of transmission to other animals, availability of vaccines

and potential zoonotic risk. This retrospective study used cases of fatal bronchopneumonia in dogs

and cats submitted to the Animal Health Lab, University of Guelph over a 5 year period to evaluate

the utility of identification of cilia-associated bacteria (CAB) on hematoxylin and eosin-stained

histologic sections to diagnose B. bronchiseptica. Detection of CAB was compared with results

from polymerase chain reaction (PCR) for flaA, immunohistochemistry (IHC) for pertatin, and

bacterial culture results. The study included 36 canine and 31 feline cases of fatal

bronchopneumonia. B. bronchiseptica was identified in 8/36 canine and 14/31 feline cases by

histological evaluation, IHC, PCR or bacterial culture. Histologic examination revealed bronchial

CAB in 4 canine and 5 feline cases; these were all positive by IHC and PCR. Of these, PCR was

positive in all cases and was the only test positive for B. bronchiseptica in 4 canine and 7 feline

cases. IHC was positive in 4 canine and 7 feline cases, positively labeling 2 additional cases not

detected with routine histologic evaluation. A subset of cases were cultured and B. bronchiseptica

was identified in 2/29 canine and 3/18 feline cases. All cases with positive bacterial culture results

had CAB identified on histologic sections. The presence of CAB appears to be a specific and useful

diagnostic feature of B. bronchiseptica pneumonia.

33

Platelet lysate as alternative to fetal bovine serum in equine/canine

mesenchymal stem cell culture

Russell, K. A., Gibson, T. W., Chong, A., Co, C., and Koch, T. G.


Mesenchymal stem cells (MSC) must be expanded to sufficient numbers in vitro for emerging

tissue regeneration therapies. Presently, expansion media (EM) for MSC is supplemented with

fetal bovine serum (FBS). However, issues with FBS are high cost, variability between batches,

and risk of complications from bovine antigens. Platelet lysate (PL) has shown promise as an

alternative to FBS in human studies.

Hypothesis: PL and FBS equally support proliferation of equine and canine MSC.

Objective: To determine how equine and canine MSC proliferate in EM enriched with PL

compared to FBS.

Proliferation assays were performed with equine cord blood-, canine adipose-, and canine bone

marrow-derived MSC in EM with 5% to 60% PL or FBS. Pooled PL was produced through a

double centrifugation method, followed by a freeze/thaw cycle. Canine cultures were assessed for

effects of long-term exposure to PL by staining for adipogenesis, chondrogenesis, and

osteogenesis.

MSC proliferated with a dose-dependent response with no significant difference found between

PL and FBS up to a 30% concentration. Beyond, proliferation fell in the PL-cultured cells, while

a continued dose-dependent proliferation response was noted in the FBS-cultured cells. Long-term

PL cultures did not support MSC expansion, and spontaneous adipogenic differentiation was

discovered even in short-term PL cultures.

We determined that PL supports MSC short-term, but not long-term. This raises questions about

whether platelet-derived products and MSC co-injected in vivo will have additive or subtractive

effects. More work is needed to determine the interactions between platelets and MSC in vitro and

in vivo.

34

A comparison of using beta-hydroxybutyrate versus clinical signs to

identify anorexic nursery pigs

Perri AM1, Friendship R1, Harding JCS2, O’Sullivan TL1

1Department of Population Medicine 2Large Animal Clinical Sciences, University of Saskatchewan

The process of weaning pigs influences structural changes in the small intestine and piglet

behaviour which can result in anorexia. Pigs that do not eat are able to maintain body condition

for approximately a week by utilizing fat stores and therefore when housed in large groups are

difficult to identify at an early stage. The clinical signs used by producers to determine if pigs are

anorexic include loss in body condition (thinness) and abnormal oral behaviour (chomping).

Elevated levels of beta-hydroxybutyrate (BHB) have been found in anorexic nursery pigs. The

objective of this study was to determine if pigs identified at 4-7 days post-weaning using clinical

signs were anorexic based on elevated BHB levels (ketosis). A total of 243 pigs were sampled

from 4 Ontario and 4 Saskatchewan farms. Thirty pigs per farm were selected based on observation

of their abnormal oral behaviour (Chomp; n=10), their poor body condition, (Thin; n=10), or their

healthy appearance (Control; n=10). Pigs with BHB values > 0.1 mmol/L were defined as ketotic.

Non-parametric analyses were used to examine differences between the groups and their BHB

values. Thin and Chomp pigs had elevated mean BHB values compared to Control pigs (P<0.001)

and (P<0.05), respectively. However, less than 15% of the pigs selected based on clinical signs

were ketotic, indicating that most were eating. In conclusion, this study found that relying on

clinical signs (thinness) and oral behaviour abnormalities (chomping) alone is not an accurate way

to identify anorexic pigs. Thus, it is necessary to use BHB testing to help identify affected animals

at an early stage and to gain a better understanding of post-weaning anorexia.

35

Mechanisms that allow oncolytic viral replication inside a tumor despite

pre-existing immunity against a virus-encoded antigen

AuYeung, W. K. A., Mould, R. C., Spangler, H., Kim, J., and W. Bridle, B. W.


Oncolytic Viruses (OV) carrying transgenes encoding tumour-associated antigens have the

ability to boost pre-existing transgene-specific T cell responses, while simultaneously mediating

direct oncolysis in the tumour. Besides improving survival, we have toxicity data that suggests the

pre-existing T cell responses against the OV-encoded transgene makes the OV safer due to

clearance of off-target infections. Nonetheless, one mystery remains unsolved: How is the OV able

to replicate and mediate oncolysis in a host with existing immunity against a virus-encoded

antigen? Two possible mechanisms are hypothesized: (1) An acute virus-induced lymphopenia

transiently removes transgene-specific T cells from the tumour environment, and (2) Tumour-

infiltrating T cells are dysfunctional due to the immunosuppressive tumor microenvironment. Our

preliminary data suggests these two events together provide a window of opportunity for the OV

to replicate in the tumour, despite the existence of virus-specific T cells. Our results show a

reduction in relative numbers of activated transgene-specific T cells in tumour and blood samples

of melanoma-bearing mice treated with the transgene-expressing OV. This suggests that boosting

with the replicating virus induced transient lymphopenia in tumours. We also compared the

percentage of transgene-specific CD8+ T cells that were capable of degranulation and production

of multiple cytokines, as a functional analysis. The data suggest the T cells found in tumors are

functionally impaired compared to those in the blood. Our results provide insight into the

mechanisms that allow viral-mediated oncolysis within hosts with existing virus-specific T cells.

36

An undescribed species of hepatozoon infecting the blood of northern

leopard frogs in Southern Ontario is uncovered using mitochondrial

genome targets.

Leveille, A. N., and Barta, J. R.


Hepatozoon spp. are heteroxenous apicomplexan blood parasites (Adeleorina: Apicomplexa)

commonly found in the erythrocytes of poikilothermic vertebrates and other hosts. Hepatozoon

species have large, thick walled, polysporocystic oocysts found in various blood-feeding

invertebrate definitive hosts. Two morphologically similar Hepatozoon species, Hepatozoon

catesbianae and Hepatozoon clamatae, infecting Ontario frogs use Culex territans (Culicidae:

Diptera) as definitive hosts. Early attempts to differentiate these parasites using sequence data

(nuclear 18S or ITS rDNA) were equivocal. Until recently, mitochondrial (mt) sequences from

adeleorinid parasites were unavailable. DNA barcoding of apicomplexan parasites has largely

focused on cytochrome b (cytb) and, more recently, cytochrome c oxidase subunit I (COI)

sequences. Our recent completion of a Hepatozoon sp. mt genome indicated that cytochrome c

oxidase III (COIII) was the most discriminating species-level locus for identifying Hepatozoon

spp. genetically. DNA was extracted from blood samples from Green (Lithobates clamitans), bull

(Lithobates catesbeianus) and northern leopard frogs (Lithobates pipiens) captured in various

locations in Ontario. A ~660bp barcode target covering the 3’ end of the mt COIII gene was PCR-

amplified and products sequenced directly. Sequences (n=23) matched H. catesbianae (n=10) and

H. clamatae (n=9), but 4 were a novel genotype only detected in leopard frogs. This study

highlights that the biodiversity of such parasites has likely been grossly underestimated because

of the inability to distinguish morphologically similar parasites in the blood of their vertebrate

hosts.

37

Raptor mortality in Ontario, Canada: a retrospective summary (1991-

2014) and the effects of west Nile virus

Smith, K., Campbell, D., Pearl, D., Salgado-Bierman, F. and Nemeth, N.


Observed mortalities in raptors and other wildlife are often associated either directly or

indirectly with human activities. As predators, raptors help maintain high biodiversity levels and

ecosystem balance across many habitats. Therefore, it is important to understand causes of

mortality among these ecologically important species. We retrospectively evaluated postmortem

diagnostic data from raptors submitted to the Ontario/Nunavut node of the Canadian Wildlife

Health Cooperative from 1991 to 2014 (n=1,404). Submissions encompassed 29 species, the most

common of which were red-tailed hawks (Buteo jamaicensi) (n=298; 21.2%) and great horned

owls (Bubo virginianus) (n=230; 16.4%). Trauma (n=703; 50.1%) accounted for the majority of

deaths among all raptor species, and was most commonly due to collision with objects such as

motorized vehicles and buildings (n=330; 46.9%), followed by emaciation (n=224; 16.0%). West

Nile virus (WNV) was the most common infectious cause of mortality (n=77; 5.5%). Data from

red-tailed hawks and great horned owls were further analyzed for the potential impacts of WNV

on mortality, including associations of demographic factors on the probability of WNV infection.

The odds of mortality due to infectious causes in red-tailed hawks were significantly greater after

the arrival of WNV to Ontario (2001-2014) compared to before (1991-2000) (Odds ratio=2.69,

p=0.0028). These results support the notion that among raptor carcasses recovered and submitted

for evaluation, human-associated deaths (e.g., due to trauma) are relatively common, and that for

some species, WNV is an important infectious cause of mortality.

38

Prevalence of Rotavirus, Astrovirus and Hepatitis E virus in Ontario rabbit

populations

Xie, X. T., Bil, J. Hammermueller, J., and Turner, P.V.


The Ontario commercial rabbit industry produces an important alternative source of meat, and

Ontario farms represent approximately 40% of Canadian operations. Production losses from birth

to weaning range up to 36%, largely from infectious enteric and respiratory diseases. In 2011, an

emerging hepatitis E virus (HEV) was discovered in commercial U.S. rabbitries with a prevalence

of 36%. Rabbit HEV is suggested to be zoonotic and is thought to contribute to rabbit enteritis

complex (REC). Rabbit rotavirus and astrovirus are also significant co-factors in REC. The

purpose of this study was to assess the prevalence of rotavirus, astrovirus, and HEV in Ontario

commercial meat rabbits and compare to other rabbit populations, including research, animal

shelters, and pets. Pooled rabbit fecal samples (n=101) were collected from 28 commercial farms,

and pooled and individual fecal samples were obtained from 7 research facilities, 4 animal shelters,

and 64 companion rabbits. Samples were extracted and PCR assays conducted to look for the

presence of rabbit-specific rotavirus, astrovirus and HEV. To date, our results show that of Ontario

commercial rabbit farms tested, 50% were positive for astrovirus and 1 was positive for rotavirus,

with no detected rabbit HEV of 101 pooled samples. Rabbit HEV was identified in 3 of 64 pet

rabbit fecal samples, with no positives for any of these 3 viruses detected to date in research or

shelter rabbit feces. These results provide a better understanding of the prevalence of viral

infections in different rabbit populations, including the potential for these viruses to cause

economic losses for industry and identification of zoonotic disease risk.

39

Thiamine degradation in commercial pet foods stored long-term at

freezing temperatures

Kritikos, G, Parr, J, Martin, B, and Verbrugghe, A

Department of Clinical Studies

Thiamine (vitamin B1) is an essential dietary nutrient required for carbohydrate metabolism

and nucleotide production. Commercial dog and cat foods are required to contain 2.25 and 5.6

mg/kg on a dry matter basis (DMB), respectively, based on requirements set forth by the National

Research Council (NRC). Little is known about the effect of freezing on thiamine degradation;

thus, the objective of this study was to determine the effect of freezing on thiamine degradation in

samples of dry dog and cat foods. Newly-manufactured samples of 3 dog foods and 3 cat foods

were tested for baseline thiamine content using HPLC, then each sample was separated into

subsamples and stored at either room temperature or at -20°C. Thiamine content was analyzed

again after 5 weeks and 3 months. The baseline thiamine content among all diets was 14.48 mg/kg

DMB. An ANOVA found significant differences in thiamine content between each of the 6 diets

(p<0.01), though there was no significant treatment-related difference (p = 0.10). Further analysis

showed no significant difference in overall thiamine content between frozen and room temperature

samples (p=0.46), though differences were present between the 5-week and 3-month time points

(p=0.03). There was no interaction between time and storage temperature (p=0.92). These results

suggest that storage times may affect thiamine content despite different storage conditions.

However, as all samples at all time points met NRC requirements for thiamine content, the clinical

significance of these findings may be minimal. A longer timeframe may provide more information

on the long-term effects of freezing on thiamine content in commercial dry pet foods.

40

Nitric oxide’s dose-dependent inhibition of uterine contractility: a

potential mechanism underlying persistent breeding-induced

endometritis in the mare

Khan, F.A.1, Chenier, T.S.1, Murrant, C.L.2, Foster, R.A.3, Hewson, J.4, and Scholtz, E.L.1

1Department of Population Medicine, 2Department of Human Health and Nutritional Sciences, 3Department of Pathobiology, 4Department of Clinical Studies, University of

Guelph.

Mares susceptible to persistent breeding-induced endometritis (PBIE) have increased uterine

nitric oxide (NO) concentrations and decreased uterine contractility. Nitric oxide may have a role

in the development of PBIE based on a hypothesized inhibitory effect on uterine contractility.

However, it is not known if NO inhibits spontaneous uterine contractility in the mare.

Reproductive tracts were collected from eight non-pregnant mares euthanized at the Ontario

Veterinary College. Using an in-vitro muscle contractility model, full-thickness uterine tissue

strips from each mare were randomly allocated to each of two groups: 1) SNAP (S-nitroso-N-

acetylpenicillamine, an NO donor); or 2) NAP (N-acetyl-D-penicillamine, vehicle and time-

matched control). These were treated at 15 min intervals with increasing concentrations (10-7 M to

10-3 M) of SNAP and NAP, respectively. Contractility data was recorded was and analyzed using

a linear mixed-effects model. A significant interaction effect of group-by-concentration was

observed. The mean contractility after treatment with 10-4 M and 10-3 M SNAP were significantly

lower than the pre-treatment contractility and the mean contractility after treatment with lower

SNAP concentrations. In contrast, contractility did not change significantly in the NAP treated

controls. These results indicate that NO has a dose-dependent inhibitory effect on spontaneous

uterine contractility in the mare. The presence of increased NO concentrations in the uteri of mares

susceptible to PBIE coupled with the findings from our study constitute a potential mechanism

underlying development of PBIE in the mare.

41

The role of microRNAs in bovine ovarian development and angiogenesis

Antunes, J. and Petrik, J.


Background: Ovarian follicle development is a tightly regulated process. MiRNAs have recently

emerged as potentially being central players in regulating the ovarian cycle. Previous studies have

identified numerous miRNAs associated with both ovarian follicular and luteal development as

well as ovarian angiogenesis. Their exact role in regulation however, still remains unclear.

Hypothesis: MiRNAs are integral to the regulation of ovarian function and coordinate ovarian

angiogenesis and follicular development, and altered miRNA expression contributes to the onset

and progression of ovarian dysfunction and reproductive disorders in the bovine species.

Objectives: 1) Identify the expression of miRNAs in bovine ovarian structures.

2) Evaluate the role of miRNAs in regulating ovarian angiogenesis and follicular and luteal

development.

Results: To date we have shown that pericytes offer a protective effect to endothelial cells through

IF. We have also shown there to be preferential miRNA expression throughout bovine follicular

development. Endothelial cells were evaluated post-VEGF/3TSR treatment and miRNAs targeting

each factor were observed to be reciprocally expressed.

Impact: The results from these studies will provide new data about the biological mechanisms that

regulate ovarian angiogenesis and follicular and luteal development. Additionally, we will

illuminate the roles of miRNAs in ovarian follicular development and angiogenesis. Although

changes in expression of pro- and anti-angiogenic factors have been described, there is currently a

significant lack of understanding of how these factors are regulated and reciprocally inhibit each

other’s expression.

42

Disease assessment of wild turkeys (meleagris gallopavo) in ontario

MacDonald, A. M., Rejman, E., Barta, J., and Nemeth, N.


Numerous disease-producing agents pose a threat to the health of both wild and domestic

animals. Since their reintroduction to Ontario in 1984, wild turkeys (Meleagris gallopavo) have

increased in number and geographic range yet information on diseases affecting them is lacking.

A retrospective analysis of wild turkey diseases diagnosed over a 20-year period at the Canadian

Wildlife Health Cooperative was performed and provides a long-term outlook of detected diseases.

Additionally, pathogens that cause disease outbreaks or decrease the fitness of commercial turkeys

(e.g., Mycoplasma spp., Eimeria spp., avian pox virus, and avian influenza viruses) were targeted

for testing in 152 hunter-harvested wild turkeys in Ontario to determine their frequency and spatial

distribution. The majority (98.7%) of individuals tested positive for ≥1 Mycoplasma spp. (of 6

identified species), 76.0% samples tested positive for Eimeria spp., 2 had avian pox skin lesions,

and 152 wild-turkeys tested negative for avian influenza viruses. Additional pathogen testing will

include Salmonella, E. coli, Campylobacter spp., Histomonas meleagridis, and

lymphoproliferative disease virus. Data generated from this research will establish baseline disease

prevalence for future monitoring of disease emergence in wild turkeys and will assess potential

pathways for disease transmission between domestic and wild turkeys. It will support emergency

management preparedness and the formation of disease prevention strategies for the Ontario

poultry sector, and contribute to conservation and management strategies to help ensure

sustainable populations of wild turkeys in Ontario.

43

Exploring pathways for achieving and sustaining good wellbeing among

Inuit in Nunatsiavut, Canada

Sawatzky, A.1, Cunsolo Willox, A.2, Harper, S.1, Shiwak, I.3, Wood, M.4, Ford, J.5, the Inuit Mental

Health and Adaptation to Climate Change team, and the Rigolet Inuit Community Government 6

1Department of Population Medicine, University of Guelph, 2Departments of Nursing, cross-appointed with Indigenous Studies, Cape Breton University, 3 'My Word: Storytelling

and Digital Media Lab', 4Department of Health and Social Development, Nunatsiavut Government, 5Department of Geography, McGill University, 6Rigolet, Nunatsiavut

Inuit across the Circumpolar North are experiencing dramatic and rapid environmental

changes, compounded by various shifts in social, cultural, and economic realms. Despite these

challenges, Inuit in Canada are actively working to maintain their culture and traditions while

advancing both individual and collective wellbeing. Improving and sustaining good wellbeing

starts with recognizing how wellbeing is conceptualized from Inuit perspectives, and further

understanding the socio-cultural contexts that support it. This case study is part of a multi-year

community-based project based in the Inuit region of Nunatsiavut, Labrador. Drawing from 100

in-depth interviews, the main goal is to explore pathways that Inuit in this region use to support

good wellbeing. The objectives are: (1) explore conceptualizations of wellbeing from the

perspectives of Nunatsiavut Inuit; (2) describe how good wellbeing is achieved and sustained; and

(3) compare pathways for good wellbeing at both individual and community levels. Nunatsiavut

Inuit consider the land to be an underlying determinant for all dimensions of wellbeing,

emphasizing its roles as healer, teacher, connector, and kin. Indeed, the land shapes and reinforces

pathways leading to cultural revitalization, supportive relationships, and a stronger sense of

community – all of which contribute to good wellbeing in Nunatsiavut. Overall, this study provides

a foundation from which to move forward and address challenges to wellbeing in changing

environments. It will help inform locally-appropriate, culturally-safe research and policy in the

Canadian North that uphold Inuit worldviews and goals for good wellbeing.

44

An investigation of the prognostic value of the TGFβ-Hippo signaling

crosstalk in canine osteosarcoma: a preliminary analysis in a tissue

microarray

Luu, A1., Schott, C.2, Mutsaers, A.1,Wood, G.2,Viloria-Petit, A.M1.

1Department of Biomedical Science, 2Department of Pathobiology

Osteosarcoma (OSA) is a commonly diagnosed bone tumor in canines. The prognosis for OSA is

poor; increasing the need for novel therapeutic targets and prognostic factors. Transforming

growth factor beta (TGFβ) and transcriptional co-activator with a PDZ-binding motif (TAZ) are

two factors well documented for their ability to promote osteoblast differentiation and cancer

progression. Thus, it is hypothesized that the levels of active TGFβ signaling and TAZ, alone or

in combination, will correlate with one or more prognostic factors of OSA progression: tumor

grade, metastasis and overall survival (OS). To address this hypothesis, a tissue microarray (TMA)

was immunolabeled to detect the levels of phosphorylated Smad2 (pSmad2), a key effector of

classical TGFβ signaling, and TAZ. The levels of pSmad2 and TAZ were scored using the Allred

method and classified as high or low by comparing to a cut-off. Fisher’s exact test was used to

seek the associations between pSmad2 or TAZ levels, and tumor grade or alkaline phosphatase

(ALP) status. Correlations between pSmad2 and TAZ levels and OS or metastasis were evaluated

using Kaplan-Meier plots and log-rank tests. Neither pSmad2 nor TAZ levels were found to be

significantly associated with tumor grade, ALP status, metastasis or OS. However, compared to

low pSmad2 or TAZ patients, patients with high pSmad2 or TAZ levels had a shorter median time

to metastasis, hazard ratio of 1.926 and 1.562, respectively. This correlation was improved when

these two markers were used in combination. These results suggest that pSmad2 and TAZ have

potential prognostic value in canine OSA and warrant further investigation with a larger data set.

45

Cartilage tissue engineering using equine cord blood-derived

mesenchymal stromal cells

Lepage, S., Kandel, R., and Koch, T.


Focal cartilage defects often precede osteoarthritis in horses. As cartilage tissue is intrinsically

slow to heal, our objective is to generate in vitro osteochondral-like plugs using equine cord blood-

derived mesenchymal stromal cells (CB-MSCs) and a bone substitute to treat focal defects. These

MSCs can be induced to form cartilage using a membrane culture system. However, this system

is unreliable, as cells can contract on the membrane and lose their chondrogenic potential.

Therefore, we hypothesize that this membrane culture system can be improved through the

inhibition of ROCK, a regulator of cell contractility. In addition, we hypothesize that

osteochondral-like plugs can be generated with CB-MSCs and a bone substitute using two

methods: a one-step method involving direct seeding of CB-MSCs onto a bone substrate, and a

two-step method involving pre-differentiation of CB-MSCs in membrane culture, then seeding the

chondrocytes onto a bone substrate. 10 CB-MSC lines were seeded on PTFE membranes and

chondrogenic media was added with or without the addition of ROCK inhibitor (ROCKi).

Approximately 50% of the cell lines failed to adhere to the membrane after chondrogenic media

was added without ROCKi, whereas 100% of the lines adhered with ROCKi. In a feasibility study,

the one-step and two-step method of osteochondral construct generation were explored with one

CB-MSC cell line. After 4 weeks, both methods produced hyaline-like cartilage on top of the bone

substrate as evidenced through histology and biochemical analysis. These promising results serve

as a prelude to further optimize the generation of these constructs to serve as a replacement for

osteochondral grafts.

46

The role of the Par6- PI3K signaling axis in TGFβ tumor suppressor

function in breast cancer and its potential for therapeutic targeting

Delroba, A. and Viloria-Petit, A. M.


Transforming growth factor beta (TGFβ) signaling promotes breast cancer metastasis, but

also inhibits tumor growth during the early stages of cancer by promoting apoptosis. We previously

observed that Par6, a key modulator of apical-basal polarity, is also a critical mediator of TGFβ-

induced apoptosis in early cancerous mammary cells. This requires Par6 phosphorylation by TGFβ

and correlates with a reduction in the activity of phosphatidylinositol 3-kinase (PI3K)/Akt. Based

on this we hypothesize that a Par6-PI3K signaling axis functions within a cell polarity-associated

signaling network that is crucial in determining cell death in response to TGFβ, and propose that

PI3K inhibition could potentially rescue TGFβ-dependent apoptosis in advanced breast cancer

cells. To address this, we first aimed at demonstrating a physical interaction between Par6 and

PI3K. We performed Par6 and p85 (PI3K regulatory subunit) immunoprecipitation and

immunoblotting on the lysates obtained from metastatic mammary cells over-expressing

exogenous flag-tagged wild type Par6 or a dominant negative mutant Par6 that cannot be

phosphorylated in response to TGFβ. Our results indicate that Par6 and p85 interact within a

complex that also include TGFβ receptor I and the small GTPase Cdc42, and that this Par6-p85

interaction is constitutive, but further enhanced by TGFβ-Par6 phosphorylation. We continue to

demonstrate this interaction and establish its requirements with endogenously expressed proteins

in human breast cancer cell lines. Our ultimate goal is to use this knowledge to develop improved

strategies to treat advanced breast cancers that rely on a functional TGFβ-PI3K interaction for

survival.

47

Epidemiology of Ixodes scapularis on pet dogs living in an emerging Lyme

disease area in Ontario

James, C. A., Pearl, D. L., Peregrine, A. S., Lindsay, L. R., and Jardine, C. M.


Invasion of the black-legged tick, Ixodes scapularis, is a growing human and animal health

concern in Ontario. Both humans and dogs are at risk of tick bites while participating in outdoor

activities, and both are susceptible to infection with pathogens transmitted by this tick including

Borrelia burgdorferi and Anaplasma phagocytophilum, the causative agents of Lyme disease and

anaplasmosis, respectively. Our objectives were to identify the species of ticks found on pet dogs

in a newly emerging area for Lyme disease in southeastern Ontario and determine if I. scapularis

ticks in this area were infected with B. burgdorferi or A. phagocytophilum. Working with 20

veterinary clinics, we collected 395 ticks, from 256 dogs, from April through August, 2015. Five

species of ticks were identified (#ticks/#dogs): I. scapularis (209/178), Ixodes cookei (28/20),

Dermacentor variabilis (152/56), Amblyomma americanum (4/3), and Rhipicephalus sanguineus

(2/2). PCR was used to detect B. burgdorferi in 14 I. scapularis from 10 dogs and A.

phagocytophilum in 3 I. scapularis from 1 dog. Ticks will be collected and PCR tested through

the fall of 2015. Using logistic regression, we will examine the associations between pet

demographics, travel history, home location and ectoparasite control and (i) the likelihood of a dog

carrying I. scapularis ticks and (ii) the odds of a removed tick being infected with B. burgdorferi

or A. phagocytophilum. This study will provide fundamental epidemiological information for

veterinarians and their clients regarding the risks associated with tick-borne diseases.

48

A comparison of exposure to risk factors for giardiasis among non-

travelers, domestic travelers and international travelers in a Canadian

community (2006-2012)

Swirski, A., Pearl, D., Peregrine, A. and Pintar, K.


The purpose of our study was to determine how demographic and exposure factors related to

giardiasis vary between travel and endemic cases. Exposure and demographic data were gathered

by public health inspectors from giardiasis cases reported from the Region of Waterloo from 2006-

2012. Logistic regression models were fit to assess differences in exposure to risk factors for

giardiasis between international travel-related cases and Canadian acquired cases while controlling

for age and sex. Multinomial regression models were also fit to assess the differences in risk

profiles between international and domestic travel-related cases and endemic cases. Travel-related

cases (both international and domestic) were more likely to go camping or kayaking, and consume

untreated water compared to endemic cases. Domestic travel-related cases were more likely to

visit a petting zoo or farm compared to endemic cases, and were more likely to swim in freshwater

compared to endemic cases and international travel-related cases. International travelers were

more likely to swim in an ocean compared to both domestic travel-related cases and endemic cases.

These findings demonstrate that travel related and endemic cases have different risk exposure

profiles which should be considered for appropriately targeting health promotion campaigns.

49

Environmental toxicants and innate pulmonary defence: the effects of

polycyclic aromatic hydrocarbons on in vitro expression of beta

defensinsin tracheal epithelial cells

Bourque, L., Raverty, S., B. Lillie and Caswell, J.


Bacterial pneumonia is an important cause of morbidity and mortality in cetaceans which may

be related to toxicant-induced suppression of respiratory immunity through exposure to oil spills

and terrestrial run-off. Polyaromatic hydrocarbons (PAHs) are constituents of fossil fuels which

are easily aerosolized and metabolized by the airway epithelial aryl hydrocarbon receptor (AHR).

AHR can interact with and affect NF-қB signalling which is the transcription factor responsible

for inducible expression of respiratory β-defensins (antimicrobial peptides). We hypothesize that

PAHs could suppress β -defensin expression in tracheal epithelial cells through AHR stimulation.

We modeled this response by measuring the effects of specific PAHs (benzo(a)pyrene (BAP) and

naphthalene) on the inducible expression of the β -defensin tracheal antimicrobial peptide (TAP)

in bovine tracheal epithelial cell culture. Primary cultures of bovine tracheal epithelial cells

(TECs) were exposed to concentrations of PAH between 0.05 to 5 µM over time, with or without

lipopolysaccharide (LPS) stimulation, and were analyzed for TAP gene expression by quantitative

RT-PCR. After 24 and 8 hours of BAP exposure, TECs had marked induction of TAP gene

expression by almost 10 fold compared to positive controls. In the case of naphthalene, TAP gene

expression was significantly suppressed at 36 hours of exposure compared to controls. Further

experiments with phenanthrene and crude oil extracts are ongoing. This data shows that PAHs

affect β -defensin gene expression, suggesting a mechanism by which environmental contaminants

can suppress innate respiratory defences and thereby lead to pulmonary disease.

50

Timp3 loss confers androgen-independence in a syngeneic mouse model

of castration-resistant prostate cancer

Pinelli, CJ., Kim, SD. and Wood, GA.


Inflammation has been linked to the development and progression of prostate cancer. Tissue

inhibitor of metalloproteinase-3 (TIMP3) is an endogenous inhibitor of inflammation, specifically

through inhibition of matrix metalloproteases (MMPs) and other enzymes, and its loss has been

linked to prostate cancer development. Our lab previously used Timp3 knockout (KO) mice

crossed into prostate-specific Pten conditional KO mice to investigate the role of Timp3 loss in

prostate cancer, and found increased inflammation and MMP activity with shorter tumor latency

in Timp3 KO prostate tumours compared to wildtype (WT). The objective of this study was to

determine if the pro-inflammatory microenvironment within Timp3 KO prostates plays a role in

the development of castration-resistant prostate cancer in an oncogene-driven mouse model. RM2

cells are murine prostate cancer cells derived from fetal urogenital sinus cells overexpressing Myc

and Ras in a C57BL/6J strain background. These syngeneic cells were injected orthotopically into

the left dorsolateral prostate lobe of Timp3 WT and KO mice, and allowed to grow for 10-12 days,

at which point mice underwent castration or sham surgery. As expected, WT mice injected with

androgen-sensitive RM2 cell tumours had significantly smaller tumours 6 days after castration

compared to sham surgery. In contrast, castrated Timp3 KO mice had larger tumours than sham

Timp3 KO mice, suggesting that Timp3 deficiency leads to castration-resistant prostate cancer.

These findings implicate heightened inflammation and MMP activity in the switch from androgen-

dependent to castration-resistant prostate cancer.

51

Microscopic Recovery of Regeneration in the Corallimorpharian,

Rhodactis sp.

Vroom, P., Peters, E. and Lumsden, J.


Corals are highly susceptible to damage as they are sessile animals. Their ability to regenerate

lost or damaged tissue is an important defense mechanism. There is little known about the

mechanics of coral regeneration and there is variability between different species. A histological

description on any corallimorpharian is lacking. We examined the microscopic morphology of

regenerating Rhodactis sp. in order to identify phases of the regeneration process as well as

determine the length of time until completion. Fragmentation of Rhodactis sp. is performed by

bisecting the animal, including the oral cavity, with a scalpel. Fifteen individuals were fragmented

on day 0 and 3 fragments were taken at the following time points: 0 and 12hr and 1, 2, 4, 8, and

14d and these were compared to 3 unharmed individuals. Corals were fixed in Bouin’s for 24hr,

paraffin embedded, cross sectioned at 4µm and stained with H&E. Patterns of tissue thicknesses

and cell densities in the healing fronts of the wound compared to normal were measured over time

however no significant differences were seen. The wound size from one healing epithelial edge to

the other did significantly decrease over time until the epithelium was continuous by 14d.

Descriptions of normal Rhodactis sp. histology as well as changes that occurred during

regeneration were described. These studies form the basis for the examination of factors like

nutrient load, light intensity, temperature and pH that may have an influence the rate of

regeneration.

52

Evaluation of the impact of a dental prophylaxis on the oral microbiota of

canines

Flancman, R. and Weese, JS.


The oral cavity houses a rich and complex bacterial population – one that likely plays

important roles in both health and disease. It has been only recently, with the advent and adoption

of next-generation sequencing, that a more complete understanding of the canine oral microbiome

exists. The longer-term effects of dental prophylaxis on the oral microbiota of dogs have not been

studied previously. The objective of this study was to evaluate the impact of a dental prophylaxis

on the oral microbiota of canines.

30 healthy dogs from a research colony had plaque samples taken just prior to, and one week

after a dental cleaning. A subset of 10 dogs was also sampled at two weeks and five weeks post-

dental cleaning.

A total of 5,984,826 sequences were obtained from 80 samples. Prior to the dental

prophylaxis, the phyla Spirochaetes (33%), Firmicutes (23%), Proteobacteria (18%), and

Actinobacteria (1%) predominated. One week after the dental cleaning, the phyla Proteobacteria

(32%) predominated, followed by Firmicutes (20%), Spirochaetes (12%), Actinobacteria (4%).

Treponema was the most commonly identified genus prior to (32%) and one week (12%) after

dental prophylaxis. One week post-dental prophylaxis, there were higher abundances of the genera

Pasteurella (1% vs. 5%), Actinomyces (0.5% vs. 2%), and Neisseria (0.4% vs. 2%) than prior to

the procedure. Jaccard analysis demonstrates community membership clusters based on time of

collection and gradually returns to the membership of the pre-dental time point at five weeks post-

dental prophylaxis.

Overall, the results indicate a diverse, but relatively stable oral microbiota in dogs.

53

Characterization of clonal mesenchymal stem cell lines derived from

equine umbilical cord blood

Haji Alizadeh, AH. and Koch, T.


Equine mesenchymal stem cells (MSCs) from umbilical cord blood are a heterogeneous group

of cells and lineage-committed progenitors, with a wide variety of regenerative properties. For a

large number of therapeutic applications, ex-vivo culture to produce adequate quantities of MSCs

is necessary. The present study investigates the morphology, proliferation and immuno-

phenotyping analysis of clonal eCB-MSCs. We employed a single cell clone culture method in

order to establish clonal cell population from equine Cord Blood-derived MSCs. The clonal

populations were grouped into fast-growing and slow-growing clones based on their proliferation

rates. The fast growing clones displayed 90-100% greater proliferation rate than the slow growing

clones. Both fast and slow-growing clones had similar cell morphology. Flow cytometry analysis

for the phenotype determination using putative MSC surface markers did not reveal any difference

between the two clonal population indicating a need for further molecular studies. This study

provides a basis for more detailed investigation to analyze differentiation potential in eCB-MSC

clones.

54

The use of network analysis for investigation of risk factors for porcine

reproductive and respiratory syndrome (PRRS)

Arruda, A G.1, Friendship, R.1, Carpenter, J.2, Hand, K.3 and Poljak, Z.1

1Department of Population Medicine, University of Guelph, 2Woodstock, ON, 3Strategic Solutions Group, Puslinch, ON

The importance of networks in the spread and control of infectious diseases has been

recognized in the last decade for different animal diseases. The objectives of this study were to

describe static relationships between swine sites and their service providers including

transportation, feed, semen, gilt and boar companies; and to extract parameters from network

analysis to be used in risk factor analysis for porcine reproductive and respiratory syndrome

(PRRS).

Data was obtained from the Ontario area regional control and elimination projects.

Demographics, biosecurity and network data were collected using a standardized questionnaire;

and disease status was obtained through historical information and on-farm sampling by herd

veterinarians. Descriptive statistics were conducted using STATA 13 and network analysis was

conducted using UCINET 6. Swine sites were divided into different clusters with specific network-

related characteristics, and these were offered as predictors in generalized linear mixed models

that had PRRS status (positive or negative) as the outcome of interest. A total of 816 swine sites

were enrolled in the study. The most fragmented network was the gilt network and the most

connected network was the feed network, which connected over 90% of all sites. Statistical

analysis showed a negative association between being part of a cluster of sites that had lower

average number of indirect connections with other sites and being positive for PRRS (OR = 0.15,

P = 0.03). Even though the number of indirect connections among sites appears to be important in

the odds of being PRRS positive, the type and frequency of contacts should be considered in future

studies.

55

RNA-seq profiling of the bronchiolar epithelium of horses with recurrent

airway obstruction: insights into pathologic response to tissue damage in

the lung

Tessier, L., Côté, O., Viel, L., Clark, ME., Diaz, A., Anders, S. and Bienzle, D .


Recurrent airway obstruction (RAO) is a chronic inflammatory lung disease affecting mature

horses similar to asthma of humans. The airway epithelium is the first defense against inhaled

airborne irritants, and impaired response is thought to be central to the pathology.

We hypothesized that expression of genes involved in inflammation, immune response and

tissue repair differs in horses with RAO exposed to an asthmatic challenge.

Changes in epithelium after exposure to an inhaled challenge were determined in horses with

and without RAO by transcriptome sequencing (RNA-seq) to identify differentially expressed

genes. Genome-based alignment and assessment of differential expression were performed with

STAR and edgeR (R), respectively. Gene set enrichment analysis following asthmatic stimulus

was also performed for both groups with Gene Set Enrichment Analysis (GSEA) software and

Search Tool for the Retrieval of Interacting Genes/Proteins (STRING).

There were 111 genes (FDR < 0.05) differentially expressed between horses with and without

RAO following an asthmatic challenge, and 81 of these were up-regulated. Genes up-regulated in

RAO horses were most significantly enriched for involvement in neutrophil and other leukocyte

migration and chemotaxis, inflammation, cell proliferation, response to wounding, coagulation and

hemostasis. Down-regulated genes suggest a difference in the circadian clock. These changes were

not observed in horses without RAO. Genes that differed most between groups and their associated

gene sets likely account for unique aspects of pathogenesis.

56

Factors associated with glucose tolerance, pre-diabetes, and type 2

diabetes in a rural community of south India: A cross-sectional study

Little, M.1, Humphries, S.2, Patel, K.3 and Dewey C1

1Department of Population Medicine, University of Guelph, 2Department of Sociology and Anthropology, University of Guelph, 3Department of International Development Studies,

Menno Simons College, University of Winnipeg

India’s national rural prevalence of type 2 diabetes has quadrupled in the past 25 years. Despite

the growing rural burden, few studies have examined putative risk factors and their relationship

with glucose intolerance and diabetes in rural areas. We undertook a cross-sectional study to

determine the prevalence of impaired fasting glucose (IFG), impaired glucose tolerance (IGT), and

type 2 diabetes in a rural area of south India. In addition, we determined which factors were

associated with type 2 diabetes. We sampled 3% of the adult population from 17 villages using a

randomized household-level sampling technique. Each participant undertook a questionnaire that

included basic descriptive information and an assessment of socioeconomic position, physical

activity, and dietary intake. Height, weight, waist and hip circumference, and blood pressure

measurements were taken. An oral glucose tolerance test was used to determine diabetes status.

We used stepwise logistic model building techniques to determine associations between several

putative factors and type 2 diabetes. 753 participants were included in the study. The age and sex-

standardized prevalence of IFG was 3.1%, IGT was 5.9%, and type 2 diabetes was 10.3%. Factors

associated with type 2 diabetes included physical activity [OR 0.81], rurality [OR 0.82],

polyunsaturated fat intake [OR 0.96], body mass index [OR 1.78], waist to hip ratio [OR 1.56],

and tobacco consumption [OR 2.67]. Our study contributes to the growing body of research

suggesting that diabetes is a significant concern in rural south India. Associated risk factors should

be considered as potential targets for reducing health burdens in India.

57

WC1+ γδ T lymphocytes from calves and heifers differentially modulate

macrophage function during early Mycobacterium avium subspecied

paratuberculosis infection.

Baquero, M. and Plattner, B.


Susceptibility for infection with Mycobacterium avium subspecies paratuberculosis (Map),

the causative agent of bovine paratuberculosis is highest in young calves and declines with age.

The percentage of γδ T lymphocytes, innate cells known to be involved during early Map infection

in calves, in peripheral blood also decreases significantly with age. The specific mechanisms by

which γδ T lymphocytes modulate the function of macrophages, the preferred host cell for Map,

during the early anti-mycobacterial immune response in calves remain poorly understood. The

objective of this study was to compare how WC1+ γδ T lymphocytes mediate macrophage function

during Map infection in calves with heifers. To achieve this objective, Map-infected monocyte-

derived macrophages (MDMs) obtained from young calves (30 – 40 days of age) and heifers (6 –

8 months of age), were co-cultured either in direct contact or not with autologous WC1+ γδ T

lymphocytes. Important mediators of host immunity against Map including nitrites, IL-17A, IFN-

γ and IL-4 from cell culture supernatants were measured. CD25 expression on γδ T cells and MHC-

I /MHC-II expression on MDMs were measured; the viability of Map recovered from MDM

cultures after Map infection was assessed. In both calves and heifers Map viability was

significantly reduced when WC1+ γδ T lymphocytes were co-cultured in contact with Map-

infected MDMs. Responses in heifers are highly variable compared to calves, however individual

variability was observed in both age groups. In conclusion, this study showed that WC1+ γδ T

lymphocytes from calves and heifers differentially modulate Map-infected MDM functions in

vitro.

58

Trends in cat admissions to Guelph animal shelter between 2011 and

2015

Janke, N., Berke, O., Klement, E., Coe, J. and Flockhart, T.


In recent years, there has been a growing concern regarding an increasing homeless cat

population, and how it can be successfully and humanely reduced. Shelters are constantly

overwhelmed by the number of cats being admitted. In 2014, the Guelph Humane Society

implemented the Capacity for Care program at its shelter. The purpose of this program is to reduce

the number of cat admissions to control for overcrowding, decrease length of stay, increase

adoption rates, and decrease euthanasia rates by improving the welfare of the cats while in the

shelter and finding alternative strategies for feral cats. Currently, there are very few studies that

assess the effectiveness of the Capacity for Care program.

In this study, newly developed time series analysis methods for count data are used to

determine if there was a significant change in the number of cats entering the shelter and their

average length of stay. Between January 2011 and September 2015, a total of 3164 cats were

admitted to the Guelph Humane Society, excluding those dead on arrival. The number of cat

admissions has decreased significantly since the Capacity for Care program was implemented. The

admissions show a strong seasonal peak over the summer months. From 2011 to 2015 the average

monthly number of cats admitted to the Guelph Humane Society decreased from 62 to 37.

In conclusion: The size of the cat overpopulation problem in Guelph has changed considerably

in recent years. Whether this is a direct effect of the Capacity for Care Program or other

confounding effects due to changes in weather or predator-prey relations needs further

investigation.

59

Clinico-pathological characterization of recently isolated Newcastle

disease virus strains in quail issing

Segovia, D., Miller, P. and Susta, L.


Newcastle disease (ND), caused by virulent strains of Newcastle disease virus (NDV), is a

devastating disease of poultry worldwide. NDV belongs to the genus Avulavirus, and contains a

negative-sense, single-stranded RNA genome 15.2 kb in length. In quail, the pathogenesis and the

potential for transmission of NDV strains is poorly understood. To characterize the ability of

virulent NDV strains to cause disease in quail, groups of 14, three-week-old Japanese quail

(Coturnix japonica) were experimentally inoculated with four highly virulent NDV strains: two

quail-isolated (N2 and N23) strains, and two poultry-isolated (Israel and Pakistan) strains. Each

bird was infected with 10^8 EID50 units of virus. At day 2 post-infection (pi), 14 non-infected

quail were mixed to each infection group (contact) to assess transmission. In inoculated quail, mild

to moderate virulence was shown, with highest mortality being 29% for N2 strain, and below 10%

for others. Only two N2-infected birds showed neurological signs typical of ND, such as head

tremor and ataxia, eyelid hemorrhages were seen in both quail-isolated strains. Microscopic lesions

were only observed in N2-, Israel-, and Pakistan-infected birds, and consisted of non-suppurative

encephalitis, gliosis, and white matter vacuolation that observed initially at day 7 through to day

10 pi. Contact birds showed no clinical signs or lesions. This study shows that virulent NDV strains

can only minimally replicate and be transmitted between quail. Furthermore, the relatively low

morbidity, mortality, and mild to moderate degrees of microscopic lesions observed in this study

suggest a certain degree of natural resistance to NDV infection.

60

Risk factors associated with stranger-directed aggression in dogs

Flint, H.1, Coe, J.1, Serpell, J.2, Pearl, DL.1 and Niel, L.1

1Department of Population Medicine, 2School of Veterinary Medicine. University of Pennsylvania

Aggression in canines is a safety concern both for humans and animals, and can lead to

decreased animal welfare in affected dogs due to abuse, neglect, relinquishment or euthanasia. Our

objective was to explore risk-factors for stranger-directed aggression in dogs using a previously

validated, owner-completed canine behaviour questionnaire (C-BARQ). Scores for stranger-

directed aggression, stranger-directed fear, non-social fear and touch sensitivity were calculated

using factors developed by Hsu & Serpell (2003) and were then dichotomized. Data were analyzed

using multivariable logistic regression models, with household as a random effect (n=16,170

dogs). This model indicated that fear of strangers (OR: 5.18; p<0.001), touch sensitivity (OR: 1.37;

p<0.001), being acquired from a pet store (OR: 1.68 vs breeder; p=0.002), being acquired from a

friend or relative (OR: 1.67 vs breeder; p<0.001) and being a neutered male (OR: 1.62 vs intact

female; p<0.001) increased the risk of being aggressive towards strangers. The age the dog was

acquired (OR: 0.87/yr; p<0.001) and the breed group were also significantly associated with

stranger-directed aggression, with hounds being the least likely (referent), and mixed breeds the

most likely to be aggressive (OR:3.58; p<0.001). The random effect of household made a

significant contribution to the model, with an ICC of 0.417, indicating the household environment

has a role in stranger-directed aggression. While a number of risk-factors for stranger-directed

aggression have been identified, our results indicate that it is important to further explore the

effects of the household environment.

61

Perioperative analgesic use by Ontario veterinarians, 2012

Reimann, J., Dewey, C., Bateman, S., Kerr, C. and Johnson, R.


Veterinarians are responsible for pain management of animals undergoing routine surgeries.

The objectives of the current study were to describe the routine use of analgesics for common

surgeries in dogs and cats by Ontario veterinarians, and to compare routine use of analgesics

between species and surgeries. Two online questionnaires, one focused on surgery in dogs, and

the other in cats, were used to gather information about veterinarians’ routine use of analgesics for

six specific surgeries in each species. Use of analgesics between species, and among surgeries,

were tested with Chi-square analyses. In total, 100 and 139 veterinarians responded to the dog and

cat questionnaires, respectively. Meloxicam, a non-steroidal anti-inflammatory agent, was the

most commonly used analgesic across the two species and all surgeries, used by 52%-79% of

veterinarians (p<0.05). The most commonly used opioid was hydromorphone, by 30% - 69% of

veterinarians (p<0.05). Between 8% and 10% of veterinarians did not report using analgesics for

dogs undergoing ovariohysterectomy and castration surgeries, respectively. Similarly, 16% and

22% of veterinarians did not use analgesics for cats for ovariohysterectomy and castration

surgeries, respectively. Greater than 60% of veterinarians reported administering analgesic drugs

pre-emptively to both dogs and cats for all surgeries. Analgesic drugs were more often dispensed

for dogs than for cats (p<0.05). Continuing education for veterinarians needs to focus on routine

use of analgesics for dogs and cats, and also on the basic understanding of pre-emptive, and multi-

modal analgesia, and the importance of dispensing analgesic drugs post-operatively.

62

Induction of innate responses in chicken embryos by administration of

encapsulated and free forms of a Toll-like receptor 4 ligand

Bavananthasivam, J., Alkie, T., Hodgins, DC. and Sharif, S.


Toll-like receptor (TLRs) are a family of innate receptors that recognize pathogen-associated

molecular patterns (PAMPs), including double-stranded RNA, CpG DNA and lipopolysaccharide.

Upon recognition of their ligand, TLRs initiate responses, manifested by activating cells and

inducing expression of cytokines that help to mediate adaptive immune responses. TLR ligands

(TLR-Ls) have the potential to be used prophylactically (alone) or as vaccine adjuvants to promote

host immunity. Further, encapsulating TLR-Ls in nanoparticles such as poly(d,l-lactic-co-glycolic

acid) may prolong responses through sustained release. Innate responses to various TLR-Ls have

been demonstrated in vitro and in vivo in chickens but have not been well studied in chicken

embryos. Therefore, in this study, encapsulated and free forms of lipopolysaccharide (LPS) (in

two different doses) were administered to embryonated eggs at 18 days of embryonation. Spleen,

lung and bursa of Fabricius were collected at 6, 18 and 48 hours post-stimulation (hps) to evaluate

expression of cytokine genes. Results suggest that free and encapsulated forms of LPS have

different effects in different tissues. In spleen, the encapsulated form of LPS upregulated interferon

(IFN)-γ, interleukin (IL)-1β and IL-10 whereas in lung and bursa of Fabricius, the free form of

LPS upregulated IL-1β, IL-10 and IFN-α, IL-1β. Significant upregulation of cytokine genes

occurred at 48 hps. Further studies are needed to evaluate the period in which encapsulated LPS

can induce protective responses against pathogens affecting young chicks.

63

An investigation of the Streptococcus suis serotype distribution in

clinically ill and healthy pigs

Arndt, E.1, Farzan, V.1, Soltes, G.2, MacInnes, J.2 and Friendship, R.1

1Department of Population Medicine, 2Department of Pathobiology

Streptococcus suis infection in pigs causes significant mortalities and production loss in swine.

Healthy carrier pigs may carry any of 35 serotypes of the microorganism. The study objective is

to investigate the serotype distribution of S. suis isolates recovered from clinical and healthy-

carrier pigs.

Sixteen farms were visited and samples collected from healthy pigs as follows: vaginal and

nasal swabs from up to 5 milking sows, nasal swabs from one of each sow’s suckling piglets, and

tonsilar swabs from up to 5 nursery and 5 grower-finisher pigs. Samples from pigs with clinical

signs include tissue samples from tonsils and/or lymph nodes, and tonsilar and meningeal swabs.

Samples were cultured and identified for S. suis at AHL. Isolates from each positive sample were

serotyped using coagglutination method.

In total, 386 samples were collected from 315 pigs (262 healthy and 53 sick). S. suis was

isolated from 67% of samples. Of 324 isolates, 181 from healthy and 40 isolates from sick pigs

were serotyped. Serotypes found in clinically ill pigs were types 8 (10%), 34 (7.5%), 4, and 23

(5% each), 2, 3, 7, 14, 22, and 25 (2.5% each); whereas serotypes from healthy carriers were types

6 ( 9.4%), 9 (4.4%), 34 (2.8%), 1, 22, and 23 (1.7% each), 14, and 19 (1.1% each), 3 and 5 (0.6%

each). In total, 94 (52%) and 15 (38%) of isolates from healthy and sick pigs were untypable,

respectively.

While the distribution of serotypes appears different between healthy and sick pigs, more sick

pigs need to be tested. A large portion of isolates were untypable or autoagglutinated by

coagglutination method and a multiplex PCR is needed to geno-serotype the isolates.

64

Mitochondrial dynamics in slow and fast growing preimplantation bovine

embryos

Merrill, C., Perkel, K. and Madan, P.


The early embryo primarily depends on mitochondria for energy production. We have

previously shown that metabolite levels differ in early embryos. Our goal therefore is to observe

mitochondrial function during the preimplantation stages of development in order to elucidate

potential molecular mechanisms behind impaired embryonic development. Firstly, In vitro

fertilized bovine embryos were randomly selected for staining with live probes TMRM

(mitochondrial membrane potential) and CM-H2DCFDA (reactive oxygen species; ROS). Time-

lapse images were obtained using a live confocal microscope equipped with an on-stage incubator.

Embryos that cleaved slower displayed higher TMRM and CM-H2DCFDA fluorescence. We then

observed embryos that were separated into slow and fast growing groups at timed stages (2-cell,

4-cell, 8-cell) and further stained with TMRM and CM-H2DCFDA. Slow growing 2-, 4- and 8-

cell embryos displayed higher TMRM and CM-H2DCFDA fluorescence than fast growing

embryos. These results indicate that slow embryos may be associated with overcompensation in

the OXPHOS pathway. High levels of ROS found in slow growing embryos may be a result of the

high mitochondrial activity. The third experiment used qPCR to determine the GLYCOX

(GAPDH, HK) and OXPHOS (ATP5b, COX5a) genes involved in developmental delay of

embryos. Slow 8-cell embryos expressed higher levels of ATP5b and both slow morula and

blastocyst embryos had high expression of GAPDH. This shows a possible compensation in slow

embryos in response to impaired mitochondrial function. Future studies will assess the effects of

OXPHOS enhancer coenzyme Q10 on gene expression.

65

Characterization of the fecal bacterial microbiota of healthy and diarrheic

calves.

Gomez, DE.1, Arroyo, LG.2, Viel, L.2 and Weese, JS1.

1Departments of Pathobiology and 2 Departments of Clinical studies

Microbiota plays a key role in health and disease, and its disruptions (dysbiosis) can trigger

diarrhea. This study aimed to characterize the fecal bacterial microbiota of healthy and diarrheic

calves.

The fecal microbiota of 20 healthy (H) and 20 diarrheic (D) calves (<4 wk) from 2 farms (F1

and F2) was characterized using the Illumina MiSeq platform targeting the V4 region of the 16S

rRNA gene. H-F1 group had higher bacterial richness than H-F2 (Chao-1, P<0.008) but no other

differences were observed. H-F1 and H-F2 were significantly different in community membership

(Jaccard index) and structure (Yu and Clayton index) (Parsimony and AMOVA, P<0.05). H and

D groups (H-F1 vs. D-F1 and H-F2 vs. D-F2) were significantly different in community

membership and structure (Parsimony and AMOVA, P<0.05). The H-F1 had higher relative

abundance of Actinobacteria (median: 22%) than D-F1 (8%) (P=0.0006), whereas H-F2 had lower

abundance of Firmicutes (44%) than D-F2 (55%) (P=0.04). LefSe analysis showed that the

Bifidobacterium genus was enriched in H groups, while Oligella, Lachnospiracea incertae sedis,

unclassified Deinoccoccales and unclassified Lachnospiracea genera were enriched in D groups.

PICRUSt analysis revealed overrepresentations of genes for tetracycline biosynthesis, metabolism

of energy, porphyrins, and lipids in D group and overrepresentations of genes for protein digestion

and absorption, vitamin B6 metabolism and biosynthesis of vancomycin group antimicrobials in

H group.

This study provides the basis for generation of hypothesis to investigate the roles that different

bacterial taxa and dysbiosis play in the development of diarrhea in calves.

66

Meteorological Drivers of Acute Respiratory Infections in Rural

Southwestern Uganda

Bishop-Williams, K.1, Sargeant, J.1, Berrang-Ford, L.2, Edge, .V1, IHACC Research Group3 and

Harper, S.1

1 Department of Population Medicine, University of Guelph , 2Department of Geography, McGill University, Montreal, 3 Indigenous Health Adaptation to Climate Change Research

Group, Montreal (J. Ford, A. Llanos, C. Carcamo, S. Lwasa, D.B. Namanya)

An estimated 1.9 million children less than 5 years old died due to acute respiratory illness

(ARI) in 2000. Rural populations in southwestern Uganda are highly climate-sensitive and may be

experiencing ARI that is linked to meteorological variables. Clinical data was collected from

Bwindi Community Hospital, from January 2011- December 2014 for children, and from

December 2011- July 2014 for adults. Daily meteorological readings for temperature and

precipitation were collected from an online repository. Logistic regression models were used to

test the associations between clinically reported ARI and demographic characteristics or

meteorological variables. A total of 27% of clinical records included a diagnosis of ARI, and

proportions were higher on the pediatric ward (36.5%). All variables significant in univariable

analysis at a lenient p-value (p<0.20) were also significant in the final multivariable model

(p<0.05). Females were 0.89 times as likely to be diagnosed with ARI than males (p=0.028), and

children aged less than 5 years were significantly more likely to be diagnosed with ARI than any

other age group (p<0.001). When controlling for age, sex, and precipitation, the odds of ARI

increased 1.08 times for every 1°C increase in daily minimum temperature (p=0.004). When

controlling for age, sex, and minimum temperature, the odds of ARI increased 1.01 times for every

1mm increase in daily precipitation (p=0.02). The proportion of ARI cases was lowest in February

(16%), and highest in August (33%). This study is the first to analyze the impacts of meteorological

drivers on ARI in rural East Africa, and illustrates a need for monitoring of ARI as climates change.

67

Bioactivity of bovine colostrum and milk exosomes and their microrna

from high, average, and low immune responders on human intestinal

epithelial cells

Ross, M.1, Atalla, H.1, 2, Gallo, N.1, Syjueco, E.1 and Mallard, B.1, 2

1Department of Pathobiology, 2Department of Animal Biosciences

Bovine milk contains bioactive components with immune-regulatory potential in humans who

consume dairy products. Expression of bioactive components in milk are often controlled by non-

coding RNA molecules known as microRNA (miRNA). Colostrum and milk miRNAs are enclosed

in exosomes, conferring their protection from degradation and potentially promoting uptake by

recipient cells. While dairy cows classified as high immune responders (HIR) have colostrum/milk

with greater immunoglobulin and beta-lactoglobulin compared to average (A) and low (L)

responders, the expression of colostrum/milk exosome-derived miRNA is unknown. Furthermore,

the bioactivity of these exosome-derived miRNA at the human intestinal epithelial barrier remains

to be explored. Therefore the purpose of this research is to evaluate the functional role of these

exosomes at the intestinal epithelial interface using human cell lines. Exosomes from the

colostrum/milk of H, A, and LIRs were isolated by differential ultracentrifugation. Electron

microscopy and immune-gold labeling of exosomes confirmed the presence of exosomes in

colostrum and milk. Additionally, exosomes were quantified by BCA protein assay and ELISA

ExoEL-CD63 kit. To assess functionality, exosomes are co-cultured with human intestinal

epithelial cells (IECs). An MTT assay is conducted to determine if exosomes are cytotoxic or

promote the viability of IECs. Uptake of PKH67 labelled exosomes by IECs is conducted to assess

the bioavailability of bovine milk exosomes in humans. Finally, transfer of exosomal miRNA to

IECs is validated using qPCR. This research will help determine the functional importance of

bovine milk in gastrointestinal health.

68

Virulence diversity of Flavobacterium psychrophilum isolates in Ontario

and correlation of bacterial numbers in fish spleen with mortality

Jarau, M., Di Natale, A., Huber, P., MacInnes, J. and Lumsden, J.


Flavobacterium psychrophilum is a Gram-negative fish pathogen, known to be the causative

agent of bacterial cold-water disease (BCWD) in freshwater-reared salmonids. This bacterium is

well recognized as the most significant cause of mortality and decreased growth of fish production

in farmed rainbow trout (Onchorhynchus mykiss) in Ontario. Although F. psychrophilum isolates

have been mostly reported to be phenotypically homogenous, knowledge on the virulence diversity

of different F. psychrophilum isolates is still lacking. Experimental trials using large numbers of

fish are required to determine virulence, however trials are laborious and time consuming. If an

endpoint, short of, but correlated to fish mortality, was available significant time and cost could

be saved. Furthermore, mortality is increasingly an unsuitable endpoint from an animal care

perspective. In this present study, we conducted preliminary infection trials using rainbow trout

juveniles to determine the virulence of 21 different F. psychrophilum isolates from our culture

collection and significant variation between isolates was noted. FPG 101 was highly virulent and

was used in a subsequent trial, in which FPG 101 was injected intraperitoneally at low (106),

medium (107) and high (108) doses in rainbow trout juveniles. Fish spleens were collected and

bacterial numbers were enumerated using quantitative polymerase chain reaction (qPCR) and

splenic bacterial colony forming units (CFUs). Results from both qPCR and splenic CFUs will be

compared with cumulative mortality to potentially allow trials to be terminated earlier.

69

Pharmacokinetics and potential toxicity of high dose meloxicam in

preweaned piglets

Viscardi, A.1, Coetzee, J.2 and Turner, PV.1

1Department of Pathobiology, 2Veterinary Diagnostic Laboratory, Iowa State University

Meloxicam is a nonsteroidal, anti-inflammatory drug licensed for use in food animals in North

America. The label dose for pigs is 0.4mg/kg, determined from studies in sows and grower pigs

experiencing chronic inflammatory pain. However, recent work has suggested that this dose may

not be efficacious when provided to piglets experiencing acute pain (for example, after castration

and tail docking) and that a higher dose may be needed. The pharmacokinetics of meloxicam have

not been studied in piglets at the age when they are routinely processed (<10 d.o.), nor have

possible toxicity effects been evaluated for an increased dose. These issues are important to ensure

drug safety at a higher dose. The objective of this study was to determine the pharmacokinetics

and possible toxicity of 1.0mg/kg meloxicam when given IM to 8 d.o. piglets. Prior to drug

administration, 5 piglets (3M, 2F) were anesthetized and instrumented with a jugular catheter for

blood collection. 24h after surgery, piglets (n=12 [6M, 6F], 5 with catheters) were given a single

IM injection of 1.0mg/kg meloxicam. Blood was collected at 0, 0.5, 1, 2, 4, 8, 12, 24, 36 and 48h

post-injection and plasma was separated and sent for meloxicam concentration analysis. 48h after

meloxicam injection, piglets were euthanized and a gross evaluation was conducted. Samples of

the stomach, liver, and kidney were taken for microscopic evaluation. Pharmacokinetic parameters

were calculated, indicating persistence of therapeutic plasma drug levels for at least 12h post-

administration. There was no gross or microscopic evidence of toxicity, suggesting that this dose

is safe to administer to piglets for analgesia evaluation.

70

The α4β2* nicotinic receptor and the developing hippocampal formation

Chung, B. and Bailey, C.


The hippocampal formation (HF) plays an important role in learning, memory and attention.

The normal development and function of this compound brain region depends on cholinergic

neurotransmission mediated by the nicotinic class of acetylcholine receptors(nAChRs). We have

recently demonstrated in mice that the α4β2* nAChR mediates direct inward currents and

facilitates excitation in principal neurons of the HF sub-region known as the cornu ammonis area

1(CA1). This signaling is developmentally regulated and is greatest during the first two weeks of

postnatal life. As the HF also comprises the cornu ammonis area 3(CA3), dentate gyrus (DG),

subiculum (SUB) and entorhinal cortex (EC), we sought to characterize α4β2* nAChR function

within the principal neurons of these sub-regions during this developmental period. Whole-cell

electrophysiological responses to acetylcholine (ACh) were recorded in principal neurons of the

CA1, CA3, DG, SUB and EC layer VI (EC-VI). We found that the magnitude of α4β2* nAChR

responses varied across HF sub-regions. Principal neurons in the SUB and EC-VI exhibited greater

inward currents and a greater rise in membrane potential when compared with neurons in the CA1,

CA3 and DG, while principal neurons in the CA1 exhibited greater inward currents when

compared with neurons in the CA3 and DG. Interestingly, we found no differences in the ability

of ACh to accelerate action potential firing frequency among these sub-regions. Our findings

demonstrate that functional α4β2* nAChRs are present in principal neurons of the HF in young

postnatal mice and that the characteristics of their responses to ACh vary across sub-regions of

this compound brain region.

71

Investigating infectious disease susceptibility in horses through targeted

next-generation resequencing of the collagenous lectin gene family

Fraser, RS., Hammermueller, JD., Meyer, A. and Lillie, BN.


Collagenous lectins are a family of soluble pattern recognition receptors that recognize

carbohydrate motifs on the surface of pathogens, and play an important role in innate immunity to

infectious disease. Genetic mutations in collagenous lectins, in various species, predispose animals

to a variety of infectious diseases. In this case-control study, we used a high-throughput, targeted

re-sequencing approach to investigate the relationship between genetic variation in equine

collagenous lectin genes and susceptibility to disease. Liver from normal (n = 35) and diseased (n

= 54) horses was obtained from animals submitted for post-mortem examination to OVC and AHL.

Animals were grouped together by diagnosis and their DNA was pooled in equal amounts, creating

a total of 24 groups containing 4-5 horses each. Targeted re-sequencing was performed on an

Illumina MiSeq and resulted in 3.4 Gb of sequence data and a mean read depth of 39x per horse.

After implementing quality control filters, we identified 2407 single nucleotide polymorphisms

(SNPs). Of these, 102 were present in the coding regions (44 missense, 58 synonymous). In silico

analysis of the missense SNPs identified 15 mutations with potential to disrupt collagenous lectin

protein structure or function. Differences in allele frequencies between the normal and diseased

populations were investigated using a Chi-square test and 18 SNPs were significantly

overrepresented in either the normal or diseased population. The SNPs discovered in this

experiment represent potential genetic contributions to disease susceptibility, and will serve as

candidates for further population-level genotyping.

72

MicroRNA involvement in estradiol-mediated synaptic plasticity

Parratt, C., LaMarre, J. and MacLusky, N.


Estrogens have long been known to have important effects on the brain with respect to

synaptic plasticity. However, the mechanisms underlying these effects remain incompletely

understood. MicroRNAs (miRs), a class of small, non-coding RNAs, have been implicated in a

number of processes in the central nervous system including differentiation and synaptic plasticity.

Estrogens influence miRNA expression in classically hormone-responsive tissues such as the

breast and uterus. It remains unknown whether estrogens also influence miRs in the brain.

MicroRNAs present a possible mechanism for the action of estrogens on synaptic plasticity. To

determine the effects of estrogens on hippocampal miRNA expression, female CD1 mice were

ovariectomized and following a period of 7-10 days were left untreated or treated with an oil

control or 3 μg/kg estradiol. Mice were euthanized 40 minutes, 6 hours or 12 hours post-injection.

Hippocampi were removed. Total RNA was extracted using the miRNeasy Mini Kit (Qiagen). 2μg

of good quality RNA from 6 hours animals was subjected to next-generation sequencing of small

RNAs. Following analysis, 4 miRNAs were shown to significantly change following estradiol

administration. Results were validated via qPCR, showing a significant difference between vehicle

and estradiol-treated animals. However, vehicle animals also showed a significant difference from

untreated controls. These results suggest an effect of injection on microRNA expression that is

prevented by estradiol treatment. Stress has been shown to decrease dendritic complexity in CA3

of the hippocampus, and this effect can be reversed by estradiol treatment.

73

Quitting a certainty for an uncertainty: The cardiac effects of bisphenol S

substitution for bisphenol A

Ferguson, M. and Pyle, WG.

Centre for Cardiovascular Research, Department of Biomedical Sciences

Bisphenol A (BPA) was used in the production of plastic products until recently. The negative

effects of BPA on health have come into focus in recent years leading to a replacement of BPA

with Bisphenol S (BPS) in many products. Like BPA, BPS is an endocrine disruptor due to the

structural similarities with 17-β estradiol, and it has been suggested that in the heart they both

signal through the Estrogen Receptor-beta (ER-beta) pathway. BPS exposure has a similar

widespread exposure as BPA and can also leach from materials. Previous studies have identified

rapid, pro-arrhythmic effects of BPS in excised mice hearts through the non-genomic disruption

of calcium handling. The effects of BPS on cardiac contractility and myofilament function are

unknown. In this study, we investigated the non-genomic effects of BPS by acutely treating

excised mice hearts with a physiologically relevant dose of BPS. We observed that BPS treatment

decreased left ventricular developed pressure by 10% over a 15 minute treatment. The decline in

contractility was driven largely by impaired systolic function. We will determine if the observed

contractile changes derive from altered actomyosin cycling and the covalent modification of

myofilament proteins. Understanding the cardiac effects of BPS is critical in determining if the

substitution of this chemical for other endocrine disruptors is safe, or if it comes with similar health

concerns.

74

The role of blood vessel formation in wound healing

Subramaniam, N., Petrik, J. and Vickaryous, M.


Wound healing is a well-characterized process that often results in scar formation. Although

scars help restore homeostasis and prevent foreign pathogen entry, they are structurally and

functionally imperfect, resulting in diminished thermoregulatory capacity, decreased tensile

strength and various psychological effects. Whereas scar formation is the most common mode of

wound healing in mammals, other species, including lizards, are capable of healing scar-free.

While many details of the scar-free wound healing process remain poorly understood, emerging

evidence indicates that blood vessel formation plays an important role. Here we test the role of

blood vessel formation during scar-free wound healing using a regenerative-competent lizard

model, the leopard gecko (Eublepharis macularius). Leopard geckos normally heal without

scarring following the creation of a full-thickness excisional skin injury. My first objective is to

alter the vascular response to wounding by inhibiting the endogenous pro-angiogenic factor

Vascular Endothelial Growth Factor (VEGF) using the small molecule inhibitor vatalanib. I predict

that treatment with vatalanib will result in a significant decrease in blood vessel formation, leading

to incomplete regeneration. My second objective seeks to exaggerate blood vessel formation at the

wound site with the exogenous application of recombinant VEGF. I predict that amplifying the

angiogenic response during wound healing will result in scar formation. These findings will be

critical in understanding the biology of scarification.

75

Role of Hippo signaling pathway during bovine preimplantation embryo

development

Sharma, J. and Madan, P.


Blastocyst formation is an important milestone during preimplantation embryo development.

During murine preimplantation embryogenesis, Hippo signaling pathway is responsible for cell

polarization thereby leading to the segregation of two distinct cell lineages during blastocyst

formation. However, the molecular mechanism of this process remains poorly understood.

Therefore, our goal is to characterize the Hippo signaling pathway during bovine preimplantation

embryogenesis and elucidate the role of this signaling pathway during blastocyst formation. Post

fertilization, bovine embryos were collected at various stages of preimplantation development (1-

cell, 2-cell, 4-cell, 8-cell, 16-cell, morula and blastocyst). Embryos were stained with MST1,

MST2 (Mammalian Sterile Twenty Like 1 and 2), TAZ (Tafazzin) and CRB3 (Crumbs 3)

antibodies, which are downstream mediators of this pathway, and visualized using laser confocal

microscopy. MST1 and MST2 proteins were localized in the cytoplasm during all the stages of

development. TAZ was localized in the cytoplasm of the embryos upto the 8-cell stage, following

which the expression shifted to the nucleus in some of the blastomeres. CRB3 proteins were

located in the cytoplasm during all the stages. Localization of MST1/2 and CRB3 demonstrates

that this pathway is active in bovine embryos. Nuclear localization of TAZ could be due to change

in polarity of the individual blastomeres as they shift from being apolar to polar. Further studies

related to regulation of Hippo signaling pathway constituents will enhance our knowledge about

compaction and cavitation and improve embryo production rates.

76

A preliminary investigation of cardiac stem/progenitor cells in the lizard

heart (Eublepharis macularius)

Jacyniak, K., Hawthorne, D., Robertson, L. and Vickaryous, M.


Resident populations of stem/progenitor cells are known to play a role in normal physiological

repair and have also been implicated to contribute to injury-mediated tissue regeneration. These

populations have been identified within various tissues across vertebrates, including the heart.

More recently, their presence has been identified within the hearts of urodeles and neonatal mice,

both reported to be capable of cardiac regeneration. Here we provide preliminary evidence of the

presence of resident cardiac stem/progenitor cells (CSPCs) in a model capable of multi-tissue

regeneration, the leopard gecko (Eublepharis macularius). Immunostaining for Proliferating Cell

Nuclear Antigen (PCNA) revealed that gecko cardiomyocytes continue to proliferate into

adulthood. Next, we performed a 5-bromo-2’-deoxyuridine (BrdU) pulse-chase experiment to

identify slow cycling cells within the normal (uninjured) heart. We found BrdU label-retaining

cells at the 20-week chase within the epicardium and ventricular myocardium and trabeculae,

indicating that quiescent or slow-cycling cells are present. Finally, we observed that select BrdU

label-retaining cells at 20 weeks were also immunopositive for the cardiac progenitor marker c-

kit. Based on these findings, we predict that putative cardiac stem/progenitor cells are present in

the gecko heart. Future research will focus on creating a puncture wound within the ventricular

wall to characterize the response of CSPCs following a direct injury to the heart.

77

The Effects of Bisphenol A on Early Bovine Embryo Development

Kahlon, N., Ferris, JC., MacLusky, N., King, WA. and Favetta, LA.


Bisphenol A (BPA) is an endocrine disrupting chemical that can have detrimental effects on

reproductive health. BPA is one of the highest volume chemicals produced worldwide and can be

found in a wide range of products. BPA was detected in over 90% of North Americans and has

been linked to infertility and reproductive failure, such as reduced in vitro fertilization success.

Expression levels of specific survival, stress, and epigenetic markers are determinants of oocytes

competence. To investigate changes correlated with BPA exposure heat shock protein (HSP70)

and DNA methyltransferase (DNMT1) were selected. HSP70 is an upstream inhibitor of apoptosis,

can be up regulated during toxic stress. DNA methylation is an epigenetic modification and

DNMT1 is the predominant form in mammals. Bovine oocytes were in vitro matured following

exposure to BPA concentrations of 0.5, 0.05 and 0.005 mg/mL and controls (no treatment and

vehicle). These concentrations represent environmentally significant levels of BPA (FDA

guidelines). Cleavage rates in the 0.5 and 0.05 mg/mL BPA group were significantly lower

compared to the controls, while no differences were detected in the lowest group. The blastocyst

rate was significantly lower in all the three treatment groups compared to controls. Western Blot

and qPCR analysis of HSP70 showed no significant differences between the treatment groups,

suggesting an alternative apoptosis pathway could be active. There were no significant differences

in DNMT1 mRNA expression, however there was a significant decrease in protein expression in

the highest group. These results suggest that oocyte exposure to BPA can compromise early

embryo development.

78

Contamination of stored drinking water and associations with acute

gastrointestinal illness in a Canadian Inuit community

Wright, C.1, Shiwak, I.2, Ford, J.3, Farahbakhsh, K.1, Edge, V.1, RICG3, IHACC Research Group4 and

Harper, SL.1

1University of Guelph, 2Rigolet Inuit Community Government, 3McGill University, 4Indigenous Health Adaptation to Climate Change Research Group

One of the highest self-reported incidence of enteric illness in global peer-reviewed literature

occurs in Inuit communities in the Canadian Arctic. This could be, in part, due to the common

practice of collecting water in plastic containers for later consumption. This research aims to

understand drinking water collection practices, risk factors for contamination, and the possible

association with self-reported acute gastrointestinal illness (AGI) in order to inform safe water

management. The study included a census survey in Rigolet, Nunatsiavut in June 2014 that

examined self-reported AGI and practices related to water, including collection, storage, and

consumption. Samples were collected from drinking water containers in homes and analyzed for

most probable number of E.coli and total coliforms. Water temperature, turbidity, and physical

traits of containers were also recorded. Prevalence of AGI during the month before the survey was

18.9% (95% CI 14.1%-24.4%), which is substantially higher than in other parts of Canada. While

treated tap water is available in homes, 83.4% (95% CI 78.0%-87.7%) of households stored water

in containers, and analysis showed that 1.2% (95% CI 0.3%-4.9%) of this water tested positive for

E. coli, and 24.7% (95% CI 18.6%-32.0%) tested positive for total coliforms. Further analysis will

explore associations between drinking water collection and storage practices, container

contamination, and self-reported AGI outcomes. The ultimate goal of the study will be to use the

generated knowledge to inform sustainable interventions, while developing the community’s

capacity to understand potential factors increasing risk of waterborne disease.

79

Investigation of the potential relationship between swine influenza virus

and Streptococcus suis infections in weanling pigs

Hopkins, D., Poljak, Z., Farzan, V., Grgić, H. and Friendship, R.


The objective of this research project is to determine if swine influenza virus predisposes

piglets to succumb to S. suis infection resulting in clinical disease and increased mortality on swine

farms. These pathogens are commonly present on swine farms and sporadic outbreaks of

streptococcal infection are an important cause of post-weaning mortality. Wang et al. (2013)

conducted a cell culture study showing increased adherence and virulence of S. suis bacteria when

swine epithelial cells were pre-infected with classical influenza H1N1 virus. Our study will explore

this research further by looking at naturally infected piglets to determine if an in-vivo relationship

between these pathogens exists. The interaction will be analyzed using a case-series study design,

where piglets will be selected if they are demonstrating clinical signs for acute meningitis caused

by S. suis infection. Cases will be selected based on clinical signs for acute meningitis and post-

mortem examinations will be conducted. Bacterial swabs from the meninges will be cultured and

S. suis isolates serotyped by co-agglutination and PCR technology to confirm the diagnosis and

differentiate between bacterial strains. Nasal swabs, blood samples will be taken from both cases

and controls to determine if there was a previous infection with influenza, confirmed using

antibody titers and laboratory testing. Finally, porcine genome sequencing of cases and controls

will be analyzed for potential genetic markers common to pigs succumbing to S.suis infection.

80

Pain assessment and management in cats and dogs by veterinarians in

southwestern Ontario

Dawson, L.1, Dewey, C.1, Stone, E.2, Guerin, M.1, Mosley, C.3 and Niel, L.1

1 Department of Population Medicine, 2 Department of Clinical Studies ,3 404 Veterinary Emergency and Referral Hospital

Effective recognition and treatment of pain is important to ensure optimal animal welfare in

veterinary patients. To explore veterinarians’ approaches to post-operative pain assessment and

management in cats and dogs, verbal interviews were conducted with 30 companion animal

veterinarians working in southwestern Ontario. Three observers independently scored interview

transcripts, and reliability and descriptive statistics were computed. Based on weighted Kappa

statistics, interview scoring had substantial inter-observer (Kw = 0.83, 0.73) and near-perfect intra-

observer (Kw = 0.92) agreement, suggesting this series of questions forms a reliable tool for

collecting pain management data in veterinary hospitals. The majority of veterinarians relied

primarily on observation of behaviour from a distance to identify post-operative pain in their

patients (60% used this for cats, 47% for dogs); only approximately 30% of veterinarians used

behavioural responses to interaction, such as reactivity to touch and palpation, which may be more

reliable than distance observations. Furthermore, few veterinarians used established pain scales

(10% used them for cats, 7% for dogs). For ovariohysterectomies, while 86% of veterinarians

provided analgesia at recommended perioperative time points, 90% did not dispense analgesics

according to recommended durations for discharged patients. Overall, results indicate that post-

operative pain assessment and management, as reported by the majority of these veterinarians, did

not follow guidelines for best practices outlined in the veterinary medical literature.

81

Evidence for early pathogen clearance in a subclinical Johne’s disease calf

model.

Stinson, KJ. and Plattner, B.


Johne’s disease (JD) is a chronic progressive enteric infection of ruminants cause by

Mycobacterium avium subsp. paratuberculosis (MAP). Cattle are infected at a young age by

ingesting MAP shed in milk or feces of an infected animal, and undergo a lengthy subclinical

periods which can last years. During this time, MAP is shed asymptomatically in the feces of the

animal. This represents a major source environmental MAP contamination and transmission within

the herd. Clinical disease manifests as chronic diarrhea, wasting, and invariably death. In Canada,

JD an estimated 30-60% of herds, with an annual economic impact of $15M. Despite this high

prevalence, it is clear that not all animals exposed to MAP develop disease, however it is unclear

if a subset of animals are resistant to infection, or become infected and subsequently clear the

disease. Calves were experimentally infected using a previously described surgical model, and

euthanized at 4, 12, 20, and 28 weeks post infection (WPI). Animals euthanized at 28 WPI

demonstrated markedly reduced lesion scores, and reduced detection of MAP from intestinal

tissues compared to calves euthanized at earlier time points. Despite residual lesions and presence

of mucosal IgG responses, 50% of these animals showed no detectable MAP, suggesting that these

animals may have cleared the infection.

82

Characterization of neural stem/progenitor cells in the brain of the

leopard gecko (Eublepharis macularius)

Macdonald, A., McDonald, R., Gilbert, E. and Vickaryous, MK.


Lizards are a promising model of amniote central nervous system regeneration. Many lizard

species are capable of regenerating a functional spinal cord following tail loss. The regenerate

spinal cord appears to arise from activated populations of neural stem/progenitor cells (NSPCs) in

the original spinal cord. The regenerative capacity of the brain is not yet well-established. To

identify whether NSPCs are present in the adult brain of the leopard gecko, we used a

bromodeoxyuridine (BrdU) pulse-chase experiment and a panel of common NSPC markers. A

population of cells bordering the ventricular system (especially the lateral ventricle) are slow-

cycling (retaining BrdU for 20 weeks) and express widely recognized NSPC markers Sox2, Sox9,

GFAP and Musashi-1. Unexpectedly, following tail loss a subset of these cells begin to proliferate

(as evidenced by PCNA immunostaining) and alter their protein expression. These findings offer

compelling evidence that endogenous NSPC populations are present in the adult lizard brain.

Furthermore, these populations appear to be responsive to distal CNS injury, although their role in

repair and regeneration is not yet known.

83

CLOCK coordinates T cell recruitment and proliferation to benefit

outcome post-myocardial infarction

Reitz, C., Tsimakouridze, E., Alibhai, F. and Martino, T.


Rationale: Myocardial infarction (MI; heart attack) initiates a temporally orchestrated

inflammatory response in the heart in order to repair damaged tissue. The role of the circadian

system in coordinating adaptive immune (T cell) responses in the heart post-MI is unknown.

Hypothesis: We hypothesize that T cells are critical to infarct healing and their responses are

coordinated by the circadian factor, CLOCK.

Methods and Results: We use our established murine myocardial ischemia/reperfusion (I/R)

model. First, I show that T cells infiltrate the infarcted myocardium and contribute to post-I/R

healing, using flow cytometry. Second, I determine that the heart-draining mediastinal lymph

nodes are the site of T cell activation post-I/R, by injection and tracking of fluorescent particles in

the myocardium. Third, I demonstrate T cell activation and proliferation in these lymph nodes

post-I/R using in vitro and in vivo techniques. Lastly, to investigate the role of the clock mechanism

in T cell responses, I use ClockΔ19/Δ19 mice which lack a functional circadian clock.

ClockΔ19/Δ19 mice show increased T cell activation in the mediastinal lymph node post-I/R

compared to wild type mice. In addition, ClockΔ19/Δ19 mice have reduced T cell recruitment to the

myocardium, coupled with increased innate immune cell infiltration. Thus, CLOCK is critical to

post-I/R infarct healing and CLOCK disruption can adversely affect outcome.

Conclusions: These studies provide the first evidence that the circadian clock coordinates T

cell responses critical for infarct healing post-I/R. Targeting circadian T cell responses can provide

a novel strategy to improve outcome in heart attack patients.

84

The protective potential of GABAergic neurosteroids against extracellular

signal-regulated kinase-induced neurotoxicity: preliminary findings

Mendell, A. and MacLusky, N.


It has been nearly 30 years since the discovery of neurosteroids in the brain. Despite extensive

research in the last three decades, the precise role of these mysterious substances in normal cellular

function remains obscure. Neurosteroid administration has been widely reported to influence

various neuronal processes, including cognition and behaviour, as well as development and aging.

In many cases, these effects have been linked to non-steroidogenic mechanisms, including

modulation of gamma-aminobutyric acid (GABA) receptor activity. While it has become apparent

that neurosteroid administration has diverse, powerful effects on the brain, the reports to date

present as many questions as answers.

Conventional sex steroids are known to exert many of their beneficial effects through the

regulation of the extracellular signal-regulated kinase (ERK) pathway. However, it has been well

documented that abnormal, prolonged activation of ERK is an indication of neuronal stress and

toxicity, and contributes to cell death. This toxic activation of ERK has been implicated in

amyloid-induced nicotinic cholinergic receptor over-activation in Alzheimer’s disease.

Recently, we discovered that the GABAergic testosterone metabolite, 5⍺-androstane-3⍺,17β-

diol (3⍺-diol), can protect against ERK activation induced by cellular stressors. Preliminary

evidence suggests that 3⍺-diol provides this protection against neurotoxic stress without disrupting

normal neuronal ERK signaling. If these neuroprotective effects extend to the abnormal, long-term

activation of ERK caused by neurotoxins such as amyloid beta protein, the implications for human

disease could be substantial.

85

A technique for determination of pulmonary artery pulse wave velocity in

horses

Silva, G.1, Guest, B.2, Gomez, DE.1, McGregor, M.2, Runciman, J.2and Arroyo, LG.1

1Department of Clinical studies and 2School of Engineering

Calcification of the tunica media of the main pulmonary arteries has been observed in a large

proportion of young racehorses. In humans, medial calcification is the most important cause of

increased arterial stiffness, and has been implicated in the pathogenesis of microvascular diseases.

Pulse wave velocity (PWV) is a marker of arterial stiffness. This study aimed to develop a

technique for determination of pulse wave velocity of the main pulmonary arteries of horses.

Seven healthy adult horses were sedated, and continuously monitored with

electrocardiography during the procedure. The pulmonary artery (PA) trunk was cannulated via

right heart catheterization, with a catheter introducer sheath (9Fr x 100cm). Introducer placement

was guided with echocardiography. A custom-made dual pressure sensor catheter (PSC) (7Fr x

170cm) was inserted through the introducer sheath, and into one of the main branches of the PA.

The position of the PSC in one of the main branches of the PA was confirmed with thoracic

radiography and pressure measurements were recorded. The time delay of the pulse waves between

the two sensors was used to calculate PWV.

The PSC placement was successfully achieved in all horses (6/6), without significant

complications, aside from transient arrhythmias. The catheter was more commonly located on the

left PA (5/6). The mean (±SD) PWV was 3.0±1.3m/s.

This study demonstrated the feasibility of a technique to determine PA-PWV in standing

horses. The technique developed may allow further investigation of the effect of calcification of

large pulmonary arteries in the development of microvascular disorders in horses.

86

Combining virotherapy with immunotherapy to treat osteosarcoma in a

preclinical and clinical model

Mould, R., Kim, J., Walsh, S., de Jong, J., Wood, G., Wootton, S., Susta, L., Petrik, J., Mustaers, T.,

and Bridle, B.


Osteosarcomas (OS) are rare forms of bone cancers that have grim prognoses. It is diagnosed

10 times more frequently in dogs than humans, with a very similar disease progression. Canine

osteosarcomas are almost uniformly fatal and survival can usually only be extended to one year

with aggressive interventions. Immunotherapy harnesses the power of a patient’s own immune

system for highly specific systemic targeting of malignant cells. Heterologous vaccination

strategies targeting tumor-associated antigens (TAA) are promising alternatives to current cancer

treatments. A replication-deficient adenovirus (human serotype 48) expressing the TAA survivin

(overexpressed by OS) will be used to prime osteosarcoma-specific T cell responses. Subsequent

delivery of an attenuated oncolytic Maraba virus (MV) expressing survivin is expected to boost

OS-specific T cells to massive numbers. An added benefit of MV is its ability to replicate in and

lyse tumor cells. This combination of oncolytic virotherapy with immunotherapy will be evaluated

in the context of a clinical trial in OS-bearing dogs. In parallel, we aim to develop a next-generation

version of this therapy in a preclinical model. It is hypothesized that IV delivery of clodronate, a

bis-phosphonate drug, to deplete macrophages, will improve several aspects of the therapy: 1.

Increase tumour-specific T cells by removing splenic marginal zone macrophages that inhibit

secondary T cell responses, 2. Increase the oncolytic viruses that reach the tumor since

macrophages remove viruses from blood, and 3. Dampen tumor-induced immunosuppression by

removing myeloid-derived suppressor cells.

87

Host defense-related proteins in bovine milk during subclinical

Staphylococcus aureus mastitis

Abdelmegid, S.1, Park, A.2, Atalla, H.3, MacKay, A.1, Caswell, J.3, Kelton, D.4 , Kirby, G.1

1 Department of Biomedical Sciences ,2 Department of Molecular and Cellular Biology, 3 Department of Pathobiology, 4

Department of Population medicine

Staphylococcus aureus is the most common contagious pathogen associated with bovine

subclinical mastitis. Current diagnosis of S. aureus mastitis is based on bacteriological culture of

milk samples and somatic cell counts (SCC) which lacks both specifity and sensitivity. Milk

contains a range of proteins of moderate or low abundance that contribute to host defense. The

objective of this study was to identify protein biomarkers of S. aureus subclinical mastitis using

gel-based and gel-free proteomic approaches. Milk samples collected from healthy control cows

(n=7) and cows that are S. aureus-positive (n=7), were fractionated to obtain the whey. Two

different proteomic approaches were used to separate and to identify milk proteins; a) In-solution

digestion followed by separation and identification of the peptides through (LC-MS/MS) and b)

(2D-DIGE) followed with in-gel digestion and identification through (LC-MS/MS). In total, 337

milk proteins were identified in both infected milk whey and control using direct LC-MS/MS with

FDR of 1.0%. Over 60% of the proteins were common to both groups, and 80 proteins were

significantly up/down regulated in S. aureus infected milk. Notably, 83 proteins were identified in

mastitic whey uniquely and associated with host defense. Furthermore, Significant differences in

protein expression detected in mastitic samples by 2D-DIGE-MS approach including

serotransferrin, polymeric immunoglobulin receptor, complement C3 and other host defense-

related proteins. To our knowledge, this is the first study that utilizes 2D-DIGE to characterize

protein expression in milk whey from cows with mastitis.

88

Seasonal variation of food security among the Batwa of Kanungu, Uganda

Patterson, K., Berrang-Ford, L., Lwasa, S., Namanya, D., Ford, J., Twebaze, F., Clark, S., Donnelly,

B., Ihacc Research Team, Harper, S.


Climate change is projected to increase the burden of food insecurity (FI) globally, particularly

among populations that depend on subsistence agriculture. The impacts of climate change will

have disproportionate effects on populations with higher existing vulnerability. Indigenous

peoples consistently experience higher levels of FI than their non-Indigenous counterparts, and are

more likely to be dependent upon land-based resources. This study aimed to understand the

sensitivity of the food system of an Indigenous African population, the Batwa of Kanungu District,

Uganda, to seasonal variation. A concurrent, mixed methods (quantitative and qualitative) design

was used. Six cross-sectional retrospective surveys provided quantitative data to examine the

seasonal variation of self-reported household FI within 10 communities. This was complemented

by qualitative data from 14 focus group discussions with community members (n=76 participants),

as well as semi-structured interviews (n=15) with local key informants, health workers and

governmental representatives. The dry season was associated with increased FI among the Batwa

in the quantitative surveys and in the qualitative interviews. During the dry season, the majority of

Batwa households reported greater difficulty in acquiring sufficient quantities and quality of food.

However, the qualitative data indicated that the effect of seasonal variation on FI was modified by

employment, wealth and community location. These findings highlight the role social factors play

in mediating seasonal impacts on FI, and support calls to treat climate associations with health

outcomes as non-stationary and mediated by social sensitivity.

89

A spatial and temporal analysis of private well water samples in Oxford

County, Canada 2005-2015

Hoffer, J., Berke, O. and Alton, G.


In Canada, more than four million residents are dependent upon private well water for

everyday use and at risk of suffering from illness caused by bacterial contamination (Jones et al.,

2006). Knowledge of the degree of contamination and burden of disease associated with

contaminated well water is limited. To determine if any change in private well water contamination

levels occurred across postal code areas, a spatial analysis was conducted on bacterial test results

derived from 24,540 voluntarily submitted private well water samples for 59 postal code areas in

Oxford County. Additionally, a space-time analysis was conducted to determine if contamination

levels changed over time and across postal code areas. Several clusters of elevated contamination

levels were identified and both geographic location and variation over time were examined.

Finally, time-series analysis was conducted to assess the impact of current educational and

promotional campaigns, on the compliance to current sampling recommendations. Through the

identification and analysis of spatial and temporal trends, specific geographic locations have been

identified as in need of particular attention when initiating public health interventions and

educational programs.

90

Disrupting the key circadian regulator CLOCK leads to age-dependent

heart disease

Alibhai, F., Tsimakouridze, E., LaMarre, J. and Martino, T.


Introduction: Circadian rhythms are important for healthy cardiovascular physiology and

molecular gene expression. However, the role of the circadian mechanism factor CLOCK in the

heart is not known.

Hypothesis: CLOCK is a key regulator of healthy cardiac growth and renewal pathways and

loss of CLOCK function causes heart disease.

Methods and Results: First, to determine the role of CLOCK in the heart we assessed male

CLOCK mutant (clockΔ19/Δ19) vs. wild type (WT) mice from 4-21 months of age. Cardiovascular

physiology was similar at 4 months; however, clockΔ19/Δ19 mice developed dilated cardiomyopathy

by 21 months evidenced by a) increased LV dilation and reduced ejection fraction

(echocardiography), b) increased fibrosis (histopathology), and c) hypotension (in vivo

hemodynamics). Next, to investigate the mechanisms underlying development of dilated

cardiomyopathy we examined CLOCK regulation of cardiac growth pathways. At 21 months

clockΔ19/Δ19 hearts had increased PI3K and WNT signalling as shown by altered miRNA

expression, increased AKT and GSK-3β phosphorylation and increased β-CATENIN abundance.

Lastly, to show that targeting the circadian mechanism can modulate heart growth, we treated 21

month mice with SR9009 (REV-ERB agonist) for 28 days. SR9009 significantly reduced heart

weight and AKT phosphorylation in WT mice. The beneficial effect of SR9009 on cardiomyocytes

will be validated in primary heart cell cultures.

Conclusion: These findings are the first to show that loss of CLOCK function causes heart

disease. This study has important implications for individuals subjected to circadian rhythm

disruption such as shift workers and individuals with sleep disorders.

91

Expression patterns of autophagy-associated proteins in canine

appendicular osteosarcoma

Schott, CR. and Wood, GA.


The majority of dogs diagnosed with appendicular osteosarcoma (OSA) eventually succumb

to pulmonary metastasis despite undergoing amputation of the affected limb, followed by adjuvant

chemotherapy. The proportion of dogs alive one year after diagnosis is improved by post-surgical

chemotherapy, but long term survival is still poor. Autophagy is a process involving cellular self-

digestion and recycling in response to adverse environmental conditions and is a survival

mechanism of normal, non-neoplastic cells. Taking advantage of this mechanism, neoplastic cells

can enhance their survival following exposure to chemotherapy. We performed

immunohistochemistry for autophagy-related proteins on a tissue microarray (TMA) composed of

naïve appendicular tumours from dogs who began standard of care treatment. Phospho-mTOR,

phospho-AKT1, phospho-S6 ribosomal protein (S6RP), Beclin-1, LC3, and p62 were examined.

There were three distinct intensities of cytoplasmic Beclin-1 labeling. Strong cytoplasmic labeling

was observed only in cases with <1 year survival (5/14) but not in cases with >1 year survival

(0/6). OSA cells exhibited a mixture of diffuse and punctate LC3 cytoplasmic labeling. Intense

phospho-S6RP labeling was identified in regions of osteoid matrix production. Phospho-mTOR

labeling was most prominent in the nuclear compartment, except for cells undergoing mitosis

which had strong cytoplasmic positivity. This preliminary study identifies Beclin-1 as a promising

candidate chemoresponsiveness-biomarker. These and other autophagy proteins will be examined

using a large canine appendicular OSA TMA (>200 cases), in search of an autophagy signature

correlated with survival.

92

Swine influenza - patterns and determinants of virus transmission in a

nursery herd

Ferreira, JB., Grgić, H., Friendship, R., Wideman, G., Nagy, Ѐ. and Poljak, Z.


The objectives of this study are to describe the dynamics of transmission of influenza in a

multi-source nursery herd and identify risk factors for recurrent infection, since classical outbreak

patterns have not been observed.

An all-in/all-out nursery barn, with piglets from 5 different sow sources, was included in the

study. Within 2 hours of arrival, 400 pigs were selected for the initial virological testing. Nasal

swabs were collected weekly for virus isolation from 81 and 75 pigs, for two trials, respectively.

Serology was performed for 8 different influenza viruses. Risk factor analysis for virological

positivity and likelihood of recurrent infection were conducted using a logistic regression model.

In study 1 all pigs were virus-positive at some point in time, with 53% being positive more

than once. At one specific moment all pigs were positive and shedding virus. Overall, 43% of pigs

were recurrently positive. In study 2 a different pattern was obtained with less than 50% of pigs

being positive. There was a significant difference in positivity of pigs among the sow sources

(p<0.01). In study 1, pigs with higher titers of heterologous maternal immunity were more likely

to have recurrent infections (p<0.01).

It was concluded that influenza virus could circulate during the nursery phase in a cyclical

pattern. Virological positivity was associated with the source herd, with high proportion of pigs

having recurrent infection. The likelihood of recurrent infections was higher for pigs with higher

level of heterologous (within-subtype) maternal immunity, which could explain ongoing issues

in the nursery phase of production.

93

Radiation sensitivity of canine melanoma cell lines: developing the

optimal radiotherapy protocol

Smich, J., Shaw, K., Poirier, V., Fleck, A. and Mutsaers, A.


In vitro assays of radiosensitivity and radiation-induced damage repair can be used to optimize

clinical radiation therapy protocols for cancer treatment. The linear quadratic model determines

radiosensitivity by generating alpha/beta ratios for each cell line. Sublethal damage repair

capability may be assessed using a split-dose response assay wherein two doses of radiation are

delivered within variable time points. Human melanoma cells have a relatively low alpha/beta

ratio, leading to the conclusion that melanoma is substantially radioresistant and coarse-

fractionated radiation protocols obtain the maximal tumour response. Consequently, clinical

practice for canine melanoma treatment also uses coarse-fractionated radiation schedules.

However, the radiosensitivity of canine melanoma has not been previously reported. The objective

of this study was to determine the alpha/beta ratio and sublethal damage repair capabilities of

canine melanoma cell lines. Four established and one novel cell line were utilized and clonogenic

survival assays were conducted. The majority of the melanoma cell lines showed substantially low

alpha/beta ratios. In addition, little impact of inter-dose time interval was observed. Together,

these results support the use of coarse-fractionated radiation therapy as the optimal treatment

approach. Future clinical investigations may further establish the optimal radiation dose and

schedule to maximize tumour response while minimizing the adverse effects on surrounding tissue.

Furthermore, the establishment and characterization of this novel canine melanoma cell line may

be useful to improve our understanding of canine melanoma biology and treatment.

94

Combined Vessel Normalization and Oncolytic Virus Therapy in the

Treatment of Advanced Stage Ovarian Cancer

Matuszewska, K., Santry, L., Petrik, J., Bridle, B. and Wootton, S.


Epithelial Ovarian Cancer (EOC) is the most lethal gynecological cancer, taking 1,750

Canadian lives per year (CCS, 2015). A ‘silent killer’, EOC fails to present obvious symptoms at

early stages, forcing diagnosis at advanced stages when current treatment strategies are largely

ineffective. This demonstrates the need for innovative approaches to combat advanced EOC.

Oncolytic virus therapy uses natural or engineered viruses, which replicate in tumor cells but leave

non-neoplastic tissues unharmed. While this may seem like a seamless tactic in the fight against

cancer, the fault in oncolytic virotherapy is the reduced tumor perfusion that results with repeated

administration of the virus. By normalizing the tumor vasculature prior to viral delivery, we aim

to increase virus infiltration, thereby optimizing viral attack on the tumor. Using an orthotopic,

syngeneic, immunocompetent mouse model of EOC, we aim to normalize the tumor vasculature

via metronomic treatment with 3TSR, a powerful regulator of angiogenesis. Subsequently, avian-

associated Newcastle Disease Virus (NDV), which causes only minor illness in humans, will be

administered as a naturally occurring oncolytic virus. For our second aim, we will engineer an

adenovirus encoding full-length murine survivin protein for the purpose of quantifying whether

3TSR-mediated vessel normalization is able to enhance delivery of vaccine-induced T-cells into

the tumor microenvironment. Successively, we will engineer NDV incorporating anti-CTLA4 to

remove the immune checkpoint blockade and enhance availability of T-cells.

95

Acknowledgments

Many thanks to the funding agencies for providing financial support for our

students and their research projects.

Blake Graham Fellowship

Canadian Wildlife Health Cooperative (CWHC),

CIHR,

Equine Guelph

IDRC

McGill University

National Science and Engineering Research Council (NSERC)

National Sciences and Engineering Research Council (NSERC)

NSERC Discovery Grant,

NSERC scholarship,

OMAFRA University of Guelph Partnership

Dairy Farmers of Ontario Doctoral Research Assistantship

Ontario Graduate Scholarship

Ontario Veterinary College Fellowship

Ontario Veterinary College

OVC Pet Trust

Public Health Agency of Canada (PHAC)

SSHRC

The Banting Research Foundation Discovery Award,

United States Department of Agriculture (USDA)

University of Guelph OVC Scholarship

96

Thanks to our judges for making the time and effort to provide positive feedback

to our student researchers:

Faculty:

Bartlewski, Pawel

Bienzle, Dorothee

Bridle, Byram

Castro, Luis

Coomber, Brenda

Favetta, Laura

Jardine, Claire

Kilkenny, Jessica

Koch, Thomas

Kulkarni, Raveendra

Lillie, Brandon

Menzies, Paula

O'Sullivan, Lynne

Petit, Alicia

Plattner, Brandon

Scholtz, Elizabeth

Sinclair, Melissa

Susta, Leonardo

Vickaryous, Matt

Students:

Abrahams, Miranda

Abdelmegid, Shaimaa

Asthana, Rashi

Balmori-Cedeno, Jaramar

Barjesteh, Neda

Bekele-Yitbareka, Alexander

Briah, Ritesh

Clow, Katie

Cooper, Carina

Day, Lindsay

Emam, Mehdi

Gilbert, Emily

Jacyniak, Kathy

Kritikos, Georgia

Laursen, Adrianna

Lin, Joanne

Louth, Emma

Milwid, Rachael

Rossi, Tanya

Santarossa, Amanda

Tatone, Elise

Tscherner, Allison

Winder, Charlot

preface - ontario veterinary college · infertility to innate immunity 2015 graduate student...

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