Proceedings and

Abstracts MU College of Veterinary Medicine presents

The 35th Annual Phi Zeta Research Day

May 4, 2012

Phi Zeta Research Day Friday

May 4, 2012 Poster Presentations: 9:30 – 10:00 a.m. 10:00 –12:15 p.m.

Adams Conference Center E-125 Vet. Med. Bldg. Interns, Residents, Grad Students, Postdocs Veterinary Professional Students

Oral Presentations: 11:00 – 1:00 p.m.

W235 Vet. Med. Bldg. (Classroom) Interns, 1st year Residents & Grad. Students

Oral Presentations: 2:30 – 3:30 p.m.

W235 Vet. Med. Bldg. (Classroom) Veterinary Professional Students

Oral Presentations: 2:30 – 3:00 p.m. 3:00 – 4:00 p.m.

W233 Vet. Med. Bldg. (Classroom) 2nd & 3rd year Residents & Grad. Students Advanced Grad. Students & Postdocs

Keynote Address: 1:00 p.m., E-126 Vet. Med. Bldg., Auditorium

“An Opportunistic Researcher’s Guide to Making Lemonade” Dr. Carolyn J. Henry, DVM, MS Professor of Oncology, Department of Veterinary Medicine/Surgery Director, Scott Endowed Program in Veterinary Oncology Interim Associate Director of Research, Ellis Fischel Cancer Center Facilitator, MU One Health/One Medicine Mizzou Advantage Initiative

Social Hour: 4:30 p.m., Adams Conference Center Atrium Initiation/Awards Banquet: 5:30 p.m., H. Richard Adams Conference Center After Dinner Presentation

“Knock knock. Who’s there? Opportunity” Dr. Carolyn Henry Co-Sponsors: College of Veterinary Medicine Dean’s Office, Office of Research, Additional Sponsor: Nestle Purina

CVM/Phi Zeta Research Day May 4, 2012 POSTER CATEGORIES H. Richard Adams Conference Center (9:30 a.m. – 10:00 a.m., Interns, Residents, Graduate Students, Postdocs) (10:00 a.m. – 12:15 p.m., Veterinary Professional Students) Veterinary Professional Students # Presenter Department Advisor 1 Sarah Bardsley VM-3 Christopher Lorson 2 Ryan Birks VM-4 Deborah Fine 3 Lynn Brockway VM-2 Philip Johnson 4 Bailey Carr VM-2 Frank Booth 5 Taylor Cragen VM-1 Rebecca Johnson 6 Anthony Dank VM-2 John Middleton 7 Naclezhada Fridman VM-2 David Kline/Eileen Hasser 8 Lauren Henderson VM-2 Tim Evans 9 John Knouse VM-3 Kimberly Selting 10 Kristin Krebs VM-3 Sandra Axiak 11 Christine Kreuzberger VM-2 Craig Franklin 12 Jeremy Mercer VM-2 Harold Laughlin 13 Nicolette Meredith VM-3 Charles Brown 14 Jacob Moskowitz VM-2 Christopher Baines 15 Jeanna Pannone VM-2 Patrick Pithua 16 Whitney Phipps/Carrissa Wood VM-3 Deborah Fine 17 Tara Piech VM-2 Rebecca Johnson 18 Sarah Pierson VM-2 Philip Johnson 19 Sarah Prochnow VM-2 Amy DeClue 20 Catherine Ruggiero VM-2 Susan Schommer 21 Rebecca Saunders VM-2 Cheryl Heesch 22 Megan Scott VM-3 Charles Brown 23 Douglas Suntrup VM-3 Michael Calcutt 24 Douglas Suntrup VM-3 John Middleton 25 Sarah Timmerman VM-2 Aaron Stoker 26 Kristin Vinson VM-2 Rebecca Johnson 27 Erin Weber VM-2 Aaron Stoker 28 Kelsey Kevin VM-2 Elizabeth Bryda 29 Kaitlin Steele VM-3 Bill Stich Interns, 1st Yr Residents & Graduate Students # Presenter Department Advisor 30 Amanda Dolgins Vet Med/Surg Shannon Reed 31 Lindsay Donnelly Vet Med/Surg Jeffrey Bryan 2nd & 3rd Yr Residents & Graduate Students 32 Rachel Scott Vet Pathobiology Charles Brown 33 Carrie Lasky Vet Pathobiology Charles Brown 36 Matthew Cook Biomed Sci/DCRC Salman Hyder Advanced Graduate Students & Postdoctoral Fellows 34 Chamisa Herrera Vet Med/Surg Jeffrey Bryan 35 Erkan Osman Vet Path/LSC Christopher Lorson 2

CVM/Phi Zeta Research Day May 4, 2012 PRESENTATION CATEGORIES Interns, 1st Yr Residents & Grad Students (11:00 a.m. – 1:00 p.m.) – W-235 Vet. Med. Classroom Moderator: Charles Wiedmeyer # Presenter Department Advisor 1 Kevin Donnelly Vet Med/Surg Elizabeth Giuliano/Rajiv Mohan 2 Pamela Fry Vet Med/Surg John Middleton 3 Kirsty Husby Vet Med/Surg Shannon Reed 4 Ignacio Idoate Vet Med/Surg John Middleton 5 Kathryn Kaufman Vet Met/Surg Tony Mann 6 Farrah Monibi Vet Med/Surg-COL James Cook 7 Carmela Pratt Vet Pathobiology Deborah Anderson

KEYNOTE ADDRESS (Vet Med Auditorium, E126 Vet Med): 1:00 – 2:00 p.m.

C O N C U R R E N T

S E S S I O N S

Veterinary Professional Students (2:30 – 3:30 p.m.), W235 Vet. Med. Classroom Moderator: Charles Wiedmeyer # Presenter Department Advisor 1 Danan Bradley VM-3 Tim Evans 2 Klay Lapa VM-2 Tony Mann 3 Jennifer Menning VM-4 Jeffrey Bryan 4 Zijin Zhou VM-2 Eileen Hasser 2nd/3rd Yr Residents & Graduate Students (2:30 – 3:00 p.m.), W233 Vet. Med. Classroom Moderator: Fred Wininger # Presenter Department Advisor 1 Carla Bermudez Vet Pathobiology Bill Stich 2 Charles Maitz Vet Med/Surg-IINMM Fred Hawthorne Advanced Grad Students and Postdoctoral Fellows (3:00 – 4:00 p.m.), W233 Vet. Med. Classroom Moderator: Fred Wininger # Presenter Department Advisor 1 Brandie Morgan Vet Med/Surg Joan Coates/Martin Katz 2 Anna Cunningham Vet Pathobiology Charles Brown 3 Glenn Jackson Vet Pathobiology Craig Franklin 4 Jacqueline Glascock Vet Path/Life Sciences Chris Lorson

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JUDGES: Veterinary Professional Students-Posters (10:00am-12:15pm, Adams) and Presentations (2:30pm-3:30pm, W235 Vet Med) (29 and 4) Name Position Department Eileen Hasser Professor Vet Bio Joan Coates Professor Vet Med and Surg Tim Evans Associate Prof. Vet Path Intern Presentations (11:00am-1:00pm, W235 Vet Med) (7) Name Position Sandra Axiak Assistant Prof. Fred Wininger Assistant Prof. Carol Reinero Associate Prof.

Department Vet Med and Surg. Vet Med and Surg. Vet Med and Surg.

Residents, Grad Students, Post Doc Posters (9:30am, Adams) (6) Name Position Kei Kurokii Assistant Prof. Kim Selting Asst. Teaching Prof. Amy DeClue Assistant Prof.

Department VMDL Vet Med Surg Vet Med and Surg

Residents, Advanced Graduate Students & Postdoctoral Fellows Presentations (2:30pm4:00pm, W233) (5) Name Position Department Craig Franklin Professor Vet Path Linda Berent Asst. Teaching Prof. VMDL Cathy Kovarik Assistant Prof. Biomed Sci

Moderators for Presentations: Intern Presentations (11:00am-1:00pm, W235 Vet Med): Chuck Wiedmeyer Veterinary Professional Students-Presentations (2:30pm-3:30pm, W235 Vet Med): Chuck Wiedmeyer Advanced Grad Students/Postdocs/Residents Presentations (2:30pm-4:00pm, W233): Fred Wininger

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ABSTRACTS

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POSTERS

TIME DEPENDENT DELIVERY OF SCAAV9-SMN IN A SMA TRANSGENIC MICE MODEL. S Bardsley, J Glascock, K George & CL Lorson (sponsor), College of Veterinary Medicine; Department of Molecular Microbiology and Immunology; Department of Veterinary Pathobiology. Spinal Muscular Atrophy (SMA) is an autosomal recessive neurodegenerative disorder found in 1:6000 individuals and is the leading genetic cause of infant mortality. SMA is caused by a homozygous deletion of Survival Motor Neuron-1 (SMN1). The clinical features of the disease are caused by specific degeneration of α-motor neurons in the spinal cord, leading to muscle weakness, atrophy and, in the majority of cases, premature death. Recent gene replacement studies have shown that using self-complementary Adeno Associated Virus serotype 9 (scAAV9-SMN) to deliver SMN to neonatal mouse models with SMA via intracerebroventricular injection (ICV) is able to rescue the phenotype. Previous studies have focused on delivering the AAV on postnatal day 2 but we would like to determine which day postnatal ICV injections of scAAV9-SMN have the greatest effect on prolonging the lifespan and improving neuromuscular function of SMA transgenic mice. Mice are genotyped postnatal day 1 and then SMA mice are injected with virus on postnatal days 2-6. Our purpose is to see how far postnatally transgenic SMA mice can be rescued. We hypothesize that earlier time point administration of AAV will work better than later time points due to the progressive loss of α-motor neurons. The project implications focus on the timing of delivery of AAV and its effects on functional recovery of the neuromuscular tissues as a possibility for future gene therapy treatments in humans. The newborn screening panel does not currently test for SMA so many individuals are not diagnosed until months or years after birth. While there is currently no cure for SMA, we hope to show that early diagnosis of SMA for severe individuals is important and necessary for future gene therapy replacement.

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SYSTEMIC INFLAMMATION IN THE HORSE: EFFECT OF RESVERATROL. LM Brockway, PJ Johnson (sponsor), JR Amorim, AR Honaker, RS Donaldson & AE DeClue, Comparative Internal Medicine Laboratory; Department of Veterinary Medicine and Surgery. Resveratrol (RES, trans-3,5,4’-trihydroxystilbene) is a phytoalexin commonly found in fruits and beverages, including red wine. Health benefits associated with the consumption of red wine have been significantly attributed to RES (the “French Paradox”). Recently, RES has been shown to exert a powerful antiinflammatory effect in laboratory animals subjected to experimental endotoxemia/sepsis. Bacterial sepsis, the systemic inflammatory response (SIR) to bacteria or their components, is a major cause of morbidity and mortality in adult horses and foals. SIR in horses significantly contributes to the pathological processes occurring in laminitis during gastrointestinal disease. The null hypothesis states that RES does not influence pro-inflammatory cytokine production in cultivated equine whole blood (Cwb) following stimulation by bacterial Pathogen Associated Molecular Patterns (PAMPs). Therefore, an in vitro Cwb study was conducted in which anti-inflammatory actions of RES were evaluated via cytotoxicity bioassay before and following inclusion of relevant PAMPs. Initial results suggested that RES had no effect on cytokine production. Measurement of other pro-inflammatory mediators is underway. Further studies are required to determine the overall effect of RES on the equine innate immune system during systemic inflammation. Evidence of an anti-inflammatory effect of RES in Cwb would provide supporting data to validate future investigations in living horses. If RES acts as an anti-inflammatory agent in horses, it could be employed during the treatment of gastrointestinal disease and help to prevent laminitis.

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V E T E R I N A R Y P R O F E S S I O N A L S T U D E N T S

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BREED-SPECIFIC VERTEBRAL HEART SIZE FOR THE DACHSHUND. RR Birks, SE Clay, LG Britt, SB Leach & DM Fine (sponsor), Department of Veterinary Medicine and Surgery. Heart disease (HD) is common in dachshunds, and is often monitored with radiographs. The radiographic vertebral heart size (VHS) was developed to objectively evaluate the size of the canine cardiac silhouette. A VHS reference range of 9.7±0.5 vertebrae was established using a mixed population of breeds. Subsequent studies have demonstrated that some breeds fall outside this reference range. We hypothesized the breed-specific VHS for normal dachshunds would not be within the generic reference range for the normal canine cardiac silhouette. Design was a retrospective and prospective cohort study. Twenty-four normal control (CON) dachshunds had thoracic radiographs (TXR) and an echocardiogram (Echo) performed. Medical records were reviewed for dachshunds with HD. Twenty-four radiographic studies from 15 dogs with a heart murmur or arrhythmia, TXR, and an Echo performed were identified. Five observers of varying levels of clinical training measured VHS for all 48 right and left lateral TXR. Measurements were performed on 2 separate days, at least 1 week apart and were recorded to the nearest 0.25 vertebra. The data was analyzed using Wilcoxon Rank-Sum analysis. The right and left VHS were correlated with Echo 2D long-axis and M-mode left atrial (LA) measurements and M-mode LA to aortic ratio. The Kappa reliability test was used to evaluate inter-observer and intra-observer variability. P11.0 vertebrae may be a reliable indicator for the presence of HD. Although HD VHS was significantly higher than CON VHS, there was overlap between groups, indicating VHS should be interpreted in conjunction with clinical and Echo findings. VHS demonstrated a strong correlation with the 2D LA echo measurement. Inter- and intra-observer variability showed substantial agreement, indicating VHS can be reliably performed by observers with varying clinical experience.

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BEHAVIORAL OUTCOMES OF SHELTER DOGS TRAINED BY COMBAT VETERANS. T Cragen, RA Johnson (sponsor), CA McKenney & S McCune, College of Veterinary Medicine; Research Center for Human-Animal Interaction (ReCHAI); WALTHAM Centre for Pet Nutrition, UK. Introduction: Each year thousands of dogs are abandoned to animal shelters due to illness, inadequate care, or behavioral issues. The latter is the most common reason for relinquishment, despite poorly behaved dogs being easily corrected with obedience training. In this study, shelter dogs were trained by combat veterans in basic behavioral commands to improve their basic behavior and adoption potential. Study Aim: To investigate the difference in behavior of shelter dogs participating in the training group with those in the control group. Methods: Dog assessments take place in the morning at the Central Missouri Humane Society. Dogs are assigned behavioral scores of 1-5 based on the animal’s ability to follow commands such as sit, stay, heel, down, leave it, and off during the Dog Obedience Skill Exam. The Exam is performed by the Dog Assessor who has been blinded to the dog’s placement in either the control group or experimental group that attends twice weekly training sessions with combat veterans. The Assessor assigns a 1-5 score during the Dog Obedience Skill Exam based on the dog’s ability to recall simple commands. A score of 1 is assigned if the dog performs immediately after the first command, a 5 is given if the dog will not perform after two commands. After data collection is complete, there will be a comparison of the behavioral scores between the control group and experimental group. Results: Data collection is ongoing. Preliminary findings will be presented on the poster. Conclusion: The intended outcome is that dogs in the experimental group that receive the optimal number of training sessions will receive better evaluations and will have improved behavior overall due to reinforcement and memory recall. Canines in the control group are likely to show poorer behavioral scores due to a lack of training. The adoption success of dogs from the treatment and control groups will be assessed in due course. This study will assess the benefit of basic obedience training in shelter dogs and its lasting effects on canine companions.

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EFFECTS OF EXERCISE ON ADIPOCTYE PHENOTYPE. B Carr & J Company (Dr. FW Booth, sponsor). Department of Biomedical Sciences. Exercise is an effective method of decreasing body fat and preventing diseases such as cardiovascular disease and Metabolic Syndrome. Increased visceral fat mass is strongly associated with disease risk, while regular exercise is associated with decreased visceral fat mass and decreased disease risk. The purpose of this study was to investigate adipose tissue phenotype, including adipocyte size, in three distinct adipose tissue depots: epicardial heart (H), omental (OM), and subcutaneous (SQ). At four months of age thirteen pigs were randomly assigned to a sedentary (SED, n=6) or aerobic exercise (EX, n=5) group. Both groups (SED and EX) were fed 800g/day of a high fat diet. Pigs underwent treatment for 16-20 weeks during which the SED pigs were restricted to their pens and EX pigs performed a moderate intensity daily aerobic exercise-training program five times a week. At four months of age and again one week prior to sacrifice, the pigs were weighed and DXA scanned. On the day of sacrifice, H, OM, and SQ adipose tissue was collected and OM adipose tissue was removed and weighed. Post-treatment body weight was higher than pre-treatment body weight in both the SED and EX pigs, but was no difference between pre- or posttreatment body weight or total OM mass between EX and SED. Within EX pigs, H adipocytes were smaller than OM and SQ and SQ was smaller than OM. Within SED pigs, H adipocytes were smaller than OM and SQ. There was no treatment effect on any of the adipose depots. We conclude that there is no effect of exercise on body weight, OM mass or adipocyte size in H, OM, and SQ. Within both groups, H adipocytes were smaller than both OM and SQ, while only in the EX group was SQ smaller than OM.

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COMPARISON OF TWO TEAT SKIN DISINFECTION TECHNIQUES. AJ Dank & JR Middleton (sponsor), Department of Veterinary Medicine and Surgery. Mastitis is defined as inflammation of the udder most often caused by a bacterial intramammary infection. The annual economic impact of mastitis on the U.S. dairy industry exceeds two billion dollars making detection and treatment essential. The National Mastitis Council recommends scrubbing the teat end with 70% isopropyl alcohol to aseptically prepare it prior to sample collection or administration of intramammary antibiotics. While efficacy of this technique for skin preparation has been studied in other species, its efficacy in the preparation of teat skin has not been studied. The objective of this study was to assess the efficacy of two methods for teat skin disinfection, iodine pre-dip versus scrubbing with an alcohol soaked pad. All four teats of 15 lactating dairy cattle at the MU dairy were assigned to treatment groups such that diagonal teats were prepared with the same technique. Diagonal treatments were alternated between odd and even-numbered cows. Treatments and samplings were performed before milking. The skin of each teat apex was sampled using a single Bacti-Swab® before and after each cleaning method. The cotton tip of each swab was aseptically severed and placed in sterile Brain Heart Infusion (BHI) media. The BHI was agitated and an aliquot was immediately removed and plated in duplicate on blood agar. When necessary, appropriate dilutions were made to facilitate bacterial colony counting. Agar plates were incubated at 37°C and total bacterial colony counts performed at 24 and 48 hours. Mean 48 hr counts were used for data analysis. Data were analyzed within and between groups using non-parametric methods (P < 0.05). Following scrubbing with alcohol or iodine, bacterial loads were significantly reduced (P < 0.001; P = 0.006, respectively). However, no significant differences were detected between groups (P > 0.487). Furthermore, neither iodine nor alcohol preparation completely eliminated bacteria from the teat end. Hence, while these techniques clean the teat they do not appear to provide complete asepsis.

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SEROTONIN (5HT) MODULATES THE HYPOXIC VENTILATORY RESPONSE. N Fridman, D Kline (sponsor), S Friskey & E Hasser (sponsor). Veterinary Research Scholars Program, College of Veterinary Medicine; Dalton Cardiovascular Research Center. Introduction: A decrease in arterial oxygen (hypoxia) activates peripheral chemoreceptors and leads to increases in breathing to maintain arterial O2. Chemosensory afferents project to the nucleus tractus solitarius (nTS) in the brainstem. The nTS also receives projections from other brain regions, including the serotonergic raphé nuclei. These projections contribute to modulation and integration of chemoafferent input to influence breathing, sympathetic nerve activity and blood pressure in response to hypoxia. We hypothesized that serotonin (5-hydroxytryptamine, 5HT) in the nTS, derived from the raphé, plays an excitatory role on the hypoxic ventilatory response (HVR). Methods: Breathing was measured by plethysmography before and after systemic or nTS manipulation of 5HT. Rats were challenged with hypoxia (12% and 8% O2, 5min each) or normoxia (21% O2, 5min), and respiratory rate, tidal volume (VT), and minute volume (volume breathed/minute) were measured. Results: Systemic (intraperitoneal, n=4) administration of methysergide, a general 5HT antagonist, had no effect on baseline breathing compared to saline vehicle. Relative to normoxia, breathing 12% and 8% O2 progressively increased respiratory rate and VT under both conditions. However, the increase in VT at 12% O2 was significantly blunted by methysergide (Δ10±3%) compared to saline (Δ27±3%). Next, 5,7-dihydroxytryptamine (5,7-DHT), a serotonin toxin, was microinjected into the nTS to ablate nTS serotonergic fibers and compared to vehicle. Seven days later, rats again were subjected to hypoxic challenges. Similar to methysergide, preliminary data (n=2) suggest a decrease in relative VT at 12% O2 in 5,7-DHT rats. The increase in VT at 12% O2 was blunted with 5,7-DHT (Δ4±6%) compared to vehicle (Δ35±6%). Future directions: Immunohistochemistry will be done to confirm ablation of serotonin fibers within the nTS and verify that catecholaminergic cells were not affected. Conclusions: With immunohistochemical verification, data suggest that serotonin in the nTS plays an excitatory role in the HVR.

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PHARMACOKINETICS AND BIOLOGICAL EFFICACY OF DOMPERIDONE IN SOUTH AMERICAN CAMELIDS. L Henderson (Dr. T Evans, sponsor), Department of Veterinary Pathobiology. PROBLEM: Maternal agalactia is a common problem encountered by newborn South American camelids (SAC). Oral administration of the D2-dopamine receptor antagonist domperidone has been used to treat agalactia in horses. However there are NO studies investigating the GI absorption /efficacy of domperidone given to SAC for the treatment of agalactia. Several medications used in horses (monogastrics) are not absorbed from GI tracts of SACs (three-compartment stomach). ANECDOTAL reports suggest that the equine dosage of 1.1 mg of domperidone/kg BW is insufficient to treat agalactic SAC, but 2.2 mg domperidone/kg BW is reportedly effective. RESEARCH OBJECTIVES AND HYPOTHESES: The purpose of this study was to determine if domperidone can be measured in SAC serum following oral administration and if there is a corresponding rise in serum concentrations of prolactin. It was hypothesized that orally administered domperidone will be absorbed and detectable in the serum of treated female SACs, resulting in increased serum prolactin concentrations. EXPERIMENTAL DESIGN AND METHODS: This study utilized a single group, crossover design in which each of four, sexually mature female alpacas received a single oral dose of domperidone (2.2 mg domperidone/kg BW). Serial blood samples were collected from an intravenous jugular catheter just before and over a 72-hour period following administration of domperidone, and serum was collected from each of these samples and frozen for later analyses for domperidone and prolactin. Using high performance liquid chromatography, it was determined that domperidone was less than the limit of detection at each time point measured. Another dose of domperidone (4.4 mg domperidone/kg BW) was administered to each alpaca, and blood samples were collected and analyzed for domperidone, as before. Serum from each time point was analyzed for prolactin using a radioimmunoassay (results pending). RESULTS: ALL measured concentrations of domperidone were less than the limit of detection for these analyses. CONCLUSIONS: Domperidone was either not absorbed or was cleared rapidly by the first-pass effect. The pending results of the prolactin analyses are critical for accurate interpretation of these experimental results. It is possible that very low concentrations of domperidone might increase prolactin concentrations. On the other hand, domperidone absorption from the SAC GI tract might not be sufficient enough to cause a rise in serum prolactin concentrations in these species..

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SERUM THYMIDINE KINASE AS A BIOMARKER FOR EARLY DETECTION OF CANCER AND MONITORING OF RESPONSE TO TREATMENT. JA Knouse & K Selting (sponsor), Department of Veterinary Medicine and Surgery. Thymidine kinase (TK) is an enzyme that plays a key role in a salvage pathway of DNA synthesis. TK exists in two forms in the mammalian body, thymidine kinase 1 (TK1) found in the cytosol and thymidine kinase 2 (TK2) found in the mitochondria. TK1 is cell cycle specific and is elevated in cells which are actively dividing or undergoing cell death. Cancers are composed of cells that are rapidly dividing and it has been noted that TK levels are elevated in individuals with certain types of cancers; such as hemangiosarcoma (HSA) and B-cell lymphoma (LSA). The purpose of this study is to examine a cancer screening tool, developed by Veterinary Diagnostics Institute, Inc. (VDI), which is designed to detect TK1 levels in the blood of dogs. Samples are being collected across the United States on a voluntary basis. The normal reference range for TK is 0 to 5.6 U/L in a healthy animal. Dogs with TK levels within the normal reference range are followed for one year using a questionnaire that is mailed to the owners. For dogs with high TK levels, thoracic radiographs and abdominal ultrasound are performed in an effort to detect occult neoplasia. In addition, TK levels are monitored over time. VDI provides test kits and information for the owner’s veterinarian to obtain blood samples, and asks that a follow-up sample be taken 30 days later if moderately elevated, or 60 days later if mildly elevated. Samples are assayed at VDI and the results are shared with the owner and their veterinarian. This study will collect and organize the data to examine the accuracy of the test. The overall goal of the project is to develop an early screening test for cancer which could lead to earlier treatment.

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MICROBIOTA DYSBIOSIS ASSOCIATED WITH DIET CHANGE IN MICE. C Kreuzberger, M Esmail, M Myles, R Livingston & C Franklin (sponsor), Research Animal Diagnostic Laboratory, Department of Veterinary Pathobiology. Determining the cause of sporadic death of research mice can be very challenging because these animals often present with minimal to no premonitory signs or pathognomonic postmortem lesions. In some cases, scattered necrosis of enterocytes associated with mild goblet cell hyperplasia is present but no known pathogen can be found. In these cases, microbiota dysbiosis is often suspected, but difficult to document, because optimal samples (i.e. samples collected before and after death) are not available. In a recent study, approximately 10% of mice fed an experimental diet high in carbohydrates and fats were found dead within 48 hours of diet change. Mice that died exhibited the aforementioned lesions of enterocyte necrosis and goblet cell hyperplasia. To determine if these deaths were associated with microbiota dysbiosis, feces from additional mice fed control or experimental diets were collected at 0, 48 and 144 hours after diet change and analyzed using Automated Ribosomal Intergenic Spacer Analysis (ARISA). Results showed that the ARISA profiles of microbiota in mice fed the experimental diet differed between 0 and 48 hours but began to normalize to 0 hour patterns by 144 hours. Differences in microbiota were further analyzed in one mouse that died 48 hours after diet change. To this end, ARISA bands that exhibited increases in intensity between 0 and 48 hours were cloned and sequenced. Preliminary results indicated that one band represented Helicobacter ganmani and the increased bacterial load from 0 to 48 hours was confirmed by real-time PCR. Cloning and sequencing of an additional band revealed that it contained multiple bacterial species closely related to Lactobacillus animalis, Lactobacillus johnsonii, and Bacteroides thetaiotaomicron. This study demonstrates that microbiota profile changes can be associated with acute death in mice experiencing changes in diet and that ARISA provides a powerful tool to monitor such changes.

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EFFICACY OF NANOPARTICULATE PACLITAXEL, CTI52010, IN A CANINE PROSTATIC CARCINOMA CELL LINE. K Krebs, A Axiak (sponsor) & S Kumar, Veterinary Research Scholars Program, College of Veterinary Medicine.

Introduction: Paclitaxel is an effective chemotherapeutic in many forms of human cancer; however, it’s formulation with Cremophor EL as a solvent causes serious side effects in dogs such as anaphylactic shock, precluding its use in veterinary clinical oncology. Nanoparticulate paclitaxel formulation CTI52010 is a novel form of paclitaxel produced without excipients or solvents, and dosages and toxicities have been determined in a phase I clinical trial in normal dogs. Currently, there is no effective method for treating prostate cancer in dogs, which is an aggressive and highly metastatic cancer. The objective of this study is to determine if the nanoparticulate paclitaxel formulation CTI52010 is effective in causing a decrease in cell viability in canine prostatic carcinoma cell line. We hypothesize that the nanoparticulate paclitaxel formulation CTI52010 will be effective at decreasing the viability of the canine prostate cells in culture in a dose-dependent manner. Materials and Methods: CTI52010 suspension in normal saline was obtained from CritiTech Inc (Lawrence, KS). Canine prostate carcinoma cell (Ace-1) viability in the presence of various concentration of the drug (5- 50 M) was performed using WST-1 assay (Roche Applied Science) according to the manufacturer’s instructions. Drug concentrations were determined using pharmacokinetic data from a phase I trial in normal dogs to utilize clinically achievable plasma concentrations. Fluorescence microscopic analysis was performed to assess the nature of microtubules in presence of the drug using antibody for  -tubulin (Cell Signaling Technology, Danvers, MA). Nucleus was counterstained with DAPI. Results: Canine prostate carcinoma cell viability decreases with increased drug concentration. A 40% decrease in viability was observed with 5 M CTI52010 treate drug concentration (up to 200 M) resulted in further 1.4 microscopy analysis revealed that the microtubule polymer mass increased with drug concentration. While at low drug concentration (15 M) le wasan observed, irregularat meshwork higher doses of microtubu (50 M) loose or compact bundles could be visualized. Conclusions: The nanoparticulate paclitaxel formulation CTI52010 was effective at decreasing the viability of the canine prostate cells in culture in a dose-dependent manner as is expected of a cytotoxic chemotherapy agent. A phase II clinical trial is indicated in dogs with prostate cancer to determine efficacy in vivo based on these results.

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EXPRESSION OF ADIPOKINES IN ADIPOSE TISSUE OF A FAMILIAL HYPERCHOLESTEROLEMIC SWINE MODEL. JT Mercer, AA Arce-Esquivel & MH Laughlin (sponsor), Department of Biomedical Sciences. Adipose tissue is a multifunctional organ that has the ability to store lipids and to secrete peptides called adipokines. Adipokines have a complex role including anti-inflammatory as well as pro-inflammatory effects. Studies suggest that anti-inflammatory adipokines (i.e. adiponectin) decrease with obesity and pro-inflammatory adipokines (i.e. resistin) increase with obesity. Inflammatory adipokines have been implicated in the pathogenesis of a number of disease states including metabolic syndrome and cardiovascular disease. PURPOSE: This study aims to determine the expression of adipokines in adipose tissue of an animal model of atherosclerosis. In addition, this study intends to evaluate the effects of exercise training on expression of these adipokines in adipose tissue. We hypothesize that exercise training would result in decreased expression of resistin, monocyte chemotactic protein-1 (MCP-1), tumor necrosis factor alpha (TNF alpha), and macrophage scavenger receptor A (SRA) whereas exercise would increase expression of adiponectin by the adipose tissue. METHODS: Sections of adipose tissue including epicardial (coronary and non-coronary), omental, and subcutaneous fat from Familial Hypercholesterolemia (FH) exercise (n=8) and sedentary (n=4) pigs were dissected and immersed in neutral-buffered 10% formalin for ≥ 24 h using standard techniques. The sections were processed to paraffin embedment, and 5 μm sections were cut with an automated microtome. Immunostaining was performed using adiponectin, resistin, TNF alpha, MCP-1, and SRA antibodies. Sections will be examined using an Olympus BX60 photomicroscope and photographed with a Spot Insight digital camera at 20X magnification. Image Pro Plus software will be used to quantify the positive area of staining. RESULTS: Currently we are in the process of collecting data from photomicrographs and analyzing data.

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ROLE OF THE MITOCHONDRIAL PROTEIN C1QBP IN THE DEVELOPMENT OF BREAST CANCER. JE Moskowitz, AM McGee & CP Baines (sponsor), Veterinary Medicine; Dalton Cardiovascular Research Center; Department of Biomedical Sciences; Department of Medical Pharmacology and Physiology. Complement 1q-Binding Protein (C1qbp) is a mitochondrial protein upregulated in many cancers including breast cancer. However, whether C1qbp plays a critical role in the development of breast cancer has not been thoroughly investigated. We therefore hypothesized that C1qbp plays a key role in the proliferation of breast cancer cells. C1qbp was overexpressed in normal human epithelial cells (MCF10A) using an adenovirus. Conversely, C1qbp was depleted in an aggressive, highly metastatic human breast cancer cell line (MDA-MB-231) using a C1qbp specific shRNA. Indices of proliferation and the expression of putative mediators of C1qbp-induced oncogenesis such as BRCA2, lipocalin, MSH, and p53, etc., were then analyzed in each of the cell lines. At this time, we have already confirmed successful overexpression of C1qbp in the MCF10A cells. Similarly we have confirmed a 90-95% knockdown of C1qbp in the shRNA-transfected MDAMB-231 cancer cells. These cells were then analyzed for the various indices described above. Preliminary data indicates increased proliferation in the C1qbp-overexpressing MCF10A cells and reduced proliferation in the C1qbp-depleted MDA-MB-231cells. Results also indicate that C1qbp downregulates the tumorsuppressor protein p53. Together, these analyses will provide novel and important insight into the role C1qbp plays in the development of breast cancer and the potential molecular mechanisms by which it does so. Such studies will hopefully lead to the development of new anti-C1qbp therapies for the treatment of breast cancer.

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SLAUGHTERHOUSE SEROSURVEILLANCE FOR CAPRINE PARATUBERCULOSIS. J Pannone & P Pithua (sponsor), Department of Veterinary Medicine and Surgery. Paratuberculosis caused by Mycobacterium avium subsp. paratuberculosis (MAP) is widely recognized as a disease that is responsible for significant economic losses across the cattle, sheep, and goat production industries. Decreased milk yield, lower reproductive efficiency, decreased growth rates, increased mortality, and early culling risks have been associated with MAP infection in cattle, yet no current data exists on the prevalence of MAP infection in goats in the United States. A seroprevalence study performed using convenience sampling of meat and dairy goats at a slaughter facility in central Missouri will provide an estimate of the prevalence of paratuberculosis by detecting MAP specific antibodies in the serum samples via ELISA testing. Individual data collected for each goat in the categories of breed, gender, and production purpose will be used to identify individual animal risk factors associated with MAP infection.

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DETERMINING MEAN ARTERIAL BLOOD PRESSURE IN SEMI-WILD LOXODONTA AFRICANA USING A CARDELL MULTIPARAMENTER MONITOR. WE Phipps, CC Wood, WK Suedmeyer & DM Fine (sponsor), Department of Veterinary Medicine and Surgery. The objective of this study was to evaluate the systolic, diastolic, and mean arterial blood pressures of working and captive free ranging African elephants (Loxodonta africana) in order to establish normal values for this species. Thirteen elephants between the ages of 2 to 28 years old of both genders (9 males, 4 females) were utilized for this study. Each elephant was examined daily before the readings were taken. The elephants were then fitted with a blood pressure cuff around the base of the tail. Their heart rates, systolic, diastolic, and mean arterial pressures, were measured via the caudal vertebral artery with a Cardell Multiparameter Monitor 9401. Each elephant was measured five times on three consecutive days except for one elephant that was measured fifteen times on a single day as training to accept the blood pressure cuff took more time. The median systolic, diastolic, and mean arterial pressures for all elephants were 178 mmHg, 131 mmHg, 147 mmHg, respectively. The median heart rate for all elephants was 50 beats per minute. Male gender, age, and inactivity significantly increased arterial pressures.

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POTENTIAL THERAPEUTIC EFFECTS OF TRAINING SHELTER DOGS ON COMBAT VETERANS WITH PTSD. TL Piech, RA Johnson (sponsor), CA McKenney & S McCune, University of Missouri College of Veterinary Medicine; Research Center for Human-Animal Interaction (ReCHAI); WALTHAM Centre for Pet Nutrition, UK. Introduction: Post-traumatic stress disorder (PTSD) is an anxiety disorder resulting from a psychologically traumatic event that has widespread prevalence in military populations. Reported incidences of PTSD are likely to be underestimated, as many veterans are reluctant to seek counseling due to the negative stigma associated with the disorder. Shelter dogs, depending on humans for training and often presenting with a high-anxiety demeanor, may provide an opportunity for the veterans to relax, exercise, and regain a sense of well-being in a community dog training environment. Study Aim: To investigate the effects of training shelter dogs on combat veteran PTSD symptoms. Methods: Investigational Review Board approval was obtained. The research team includes a psychologist. Veterans were randomly assigned to a control or treatment group. Veterans in the treatment group were paired with a shelter dog and are participating in a dog walking and obedience training program for one hour, twice per week over 24 weeks. The training program involves teaching the dogs commands and behavioral skills with the aim of increasing adoption rate and reducing adoption relinquishment. A nationally validated, reliable PTSD Checklist- Military Version and blind evaluation were used to score veterans on specified PTSD symptom parameters pretest, midtrial, and posttest. Evaluations completed by the veterans in the treatment group after each training session were assessed for possible indications of reduction in PTSD symptoms. Results: Six veterans in the treatment group are participating in the training program. Data collection is ongoing. Preliminary findings will be presented on the poster. Veteran evaluation comments, such as "I have had a busy day at work, and reiterate that when I have exposure to a dog, I feel much better" and "The dog exhibited marked improvement in all phases of training” suggest that the study is benefiting both the shelter dogs and veterans. Conclusions: This study may encourage development of PTSD support programs that are beneficial for both veterans and shelter dogs.

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EFFECT OF TLR2 SIGNALING DEFICIENCY ON THE CHEMOKINE RESPONSE TO A BORRELIA BURGDORFERI INFECTION. NR Meredith, A Ritzman & CR Brown (sponsor), Department of Veterinary Pathobiology. Lyme disease is a tick-transmitted inflammatory disorder caused by the spirochete, Borrelia burgdorferi. Inflammatory diseases such as arthritis and carditis are associated with Lyme disease. Arthritis in mice is first detectable 1-2 weeks after infection, peaks at 4 weeks and then resolves over the next several weeks. B. burgdorferi expresses several lipoproteins that activate TLR2 and it was once thought that mice deficient in TLR2 would not develop an inflammatory response to this pathogen. However, TLR2 knockout mice infected with B. burgdorferi actually developed a worsened form of arthritis and had 50 times more spirochetes present in joint tissues. Other TLRs are not thought to compensate for the loss of TLR2, since mice deficient in the common signaling adapter, MyD88, also developed arthritis in this model. Since TLR2 knockout mice developed an exacerbated inflammatory response following B. burgdorferi infection, we wanted to determine if there was a difference in chemokine production in the joints between TLR2 knockout and wild type C3H mice. The mice were infected by injecting B. burgdorferi into the hind footpads and groups of five mice were sacrificed weekly for four weeks. Ankle tissues were homogenized and levels of chemokines were quantified by ELISA.

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STRATIFIN EXPRESSION IN THE HOOF LAMELLAR INTERFACE OF LAMINITIC HORSES. SR Pierson, CE Wiedmeyer, A Culp, DY Kim, TJ Reilly & PJ Johnson (sponsor), Department of Veterinary Medicine and Surgery. Background: Laminitis is an extremely painful and debilitating disease of horses. Results of a proteomics analysis demonstrated that stratifin is differentially expressed in laminitic hoof lamellar interface (HLI) tissue. Stratifin is a keratinocyte-derived anti-fibrogenic factor, also known as 14-3-3, with potent MMP-1 stimulatory effects on proximate dermal fibroblasts. Abnormal stratifin production could contribute to extracellular matrix (ECM) dysregulation, a well-recognized component of laminitis in horses. Moreover, stratifin promotes keratinocyte cell survival (inhibitor of apoptosis) and is important for regulation/organization of the cytoskeleton. Therefore, stratifin may be important because elevated protease-mediated ECM degradation, increased keratinocyte apoptosis, and keratinocyte dysadhesion resulting from defective hemidesmosome/cytoskeletal association have all been demonstrated in the pathogenesis of laminitis. Objective: • Determine if a difference exists regarding total stratifin expression in HLI tissue from laminitic (acute and chronic) and unaffected horses using Western blot (WB) analysis. • Determine if a difference exists regarding stratifin localization in the HLI using immunohistochemistry(IHC). Results: Differential expression of stratifin between normal and laminitic HLI tissues was not detected using WB or IHC. Clinical importance: Results of these studies failed to demonstrate abnormality of stratifin in laminitis.

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EFFECTS OF OPIOIDS ON PHAGOCYTIC FUNCTION, OXIDATIVE BURST CAPACITY AND APOPTOSIS IN DOGS. S Prochnow, DH Yu, S Axiak, J Dodam & AE DeClue (sponsor), Department of Veterinary Medicine and Surgery. Opioids alter immune responses and apoptotic pathways during sepsis in multiple species. Although, opioids are commonly used as analgesic drugs in dogs with sepsis, immunomodulatory and apoptotic effects these drugs are largely unknown in this species. The objective of this study was to evaluate the exvivo effect of morphine, fentanyl or buprenorphine on phagocytic function, oxidative burst production capacity and lymphocyte apoptosis in dogs. Blood from six healthy, adult dogs was incubated with morphine (200 ng/mL), fentanyl (10 ng/mL), buprenorphine (10 ng/mL) or control (saline) for 3 (phagocytic function) or 8 (apoptosis) hours. Neutrophil phagocytosis of opsonized E. coli, respiratory burst capacity after opsonized E. coli or phorbol 12-myristate 13-acetate (PMA) stimulation, and annexin binding to apoptotic lymphocytes were evaluated using flow cytometry. Data were compared using RMANOVA or Friedman’s test and Student-Newman-Keuls post hoc analysis (p