Master of Science / Department of Biomedical Sciences / Sanjeev K. Narayanan / Cefovecin is an extended-spectrum long-acting third generation cephalosporin used to treat canine infections. The study objective was to determine the effect of cefovecin on the absolute number and antimicrobial susceptibility of fecal enteric bacteria in healthy dogs. Fourteen Beagles were randomly assigned to a treated (n = 7, 8 mg/kg cefovecin subcutaneously on day 1) or untreated (n = 7) group. LC/MS was used to determine plasma cefovecin concentration on day 14. E. coli, enterococci, and Salmonella were isolated and enumerated from fecal samples collected on days 0, 3, 7, 14, and 28. Antimicrobial resistance was determined using disc diffusion, MIC, and detected using PCR for the bla[subscript CMY-2] gene on select isolates. Mean plasma concentration of cefovecin on day 14 was 9.59 µg/mL in treated dogs; untreated dogs had no measurable plasma cefovecin. The absolute number of E. coli was lower in treated dogs on day 3 (P ≤ 0.0001), and the absolute number of cefovecin-resistant E. coli was higher in treated dogs on days 7 (P = 0.002), 14 (P = 0.004) and 28 (P ≤ 0.0001), compared to untreated dogs. Enterococci increased and were higher in the treatment group on day 7 (P = 0.0226). Isolation of Salmonella was rare. After cefovecin treatment, beta-lactam resistance was more common in fecal E. coli from treated dogs than untreated dogs, while resistance of enterococci was not altered. On day 28, treated dogs were 3.25 times more likely to carry the bla[subscript CMY-2] gene than untreated dogs (95% CI 1.27 – 8.35). The implications of these findings in clinically ill patients require further research.

Master of Science / Department of Clinical Sciences / Amy Rankin / Objective- To evaluate the use of a topical ophthalmic steroid (1% prednisolone acetate) and non-steroidal anti-inflammatory drug (0.1% diclofenac) on blood glucose concentrations, serum fructosamine concentrations, and clinical scores in diabetic dogs with cataracts using descriptive analysis. Animals- Twelve client-owned dogs with naturally-occurring, controlled (per history and physical examination), insulin-treated diabetes mellitus and cataract. A total of 40 dogs will be enrolled in the study, as determined by power analysis. Procedures- This was a prospective, randomized, double-masked, experimental study with 2 phases of data collection. Dogs were enrolled from October 2011 to March 2014 and were assigned to 1 of 2 treatments (Drug Red or Drug Blue) using blocked randomization; dogs received either 1% prednisolone acetate suspension or 0.1% diclofenac solution. Patient history, physical, and ophthalmic examinations were performed and a clinical score assigned at enrollment (Phase 1 [day 0]) and upon return (Phase 2 [day 32]). At these times, a complete blood count, serum chemistry, urinalysis, and serum fructosamine concentration were performed prior to hospitalization for up to 72 hours for continuous glucose monitoring. For 4 weeks (day 3 to 31), dogs returned home, and owners administered the dispensed ophthalmic medication 4 times daily to both eyes. Descriptive analysis of data was performed; statistical analysis will follow enrollment of 40 dogs. Results- Twelve dogs have completed the study, with 6 dogs assigned to each treatment group. Dogs received 4.44 or 0.44 mg/day of prednisolone acetate or diclofenac, respectively. Dogs assigned to Drug Red more commonly exhibited elevations in serum liver enzyme activity. Drug Red group showed a greater percent increase in fructosamine concentrations over time. Based on glucose curves alone (22 curves analyzed), an insulin dose increase was recommended for 12 curves. An insulin dose decrease and no dose change were recommended for 5 curves each. During treatment, 1 dog reportedly developed polyuria and polydipsia. Conclusions- Descriptive analysis revealed differences in some outcomes of interest among dogs treated with 2 different ophthalmic anti-inflammatory medications. Data collection is ongoing to determine if statistically significant differences exists for outcomes per group.

Master of Public Health / Department of Diagnostic Medicine and Pathobiology / M. M. Chengappa / Rabies is the most deadly disease on earth and has a 99.9% human fatality rate. Rabies kills 61,000 humans annually and results in an economic burden of $124 billion USD annually. Each day 3.3 million people live with the risk of rabies. It is estimated that 95% of human rabies cases are a result of coming in contact with an infected canine, majority of these cases being children 15 years and younger. It is estimated that 1 person every 8 minutes dies of rabies. Rabies is a highly neurotropic disease which attacks the brain and central nervous system. Once clinical symptoms are presented, death is invariably the outcome as no cure exists for rabies. Rabies is 100% preventable in humans by proper wound management and proper administration of prophylaxis. Rabies can be adequately controlled in animal populations by contraception and animal rabies vaccine efforts. Whilst it is known that rabies can be prevented in humans and controlled in animal populations, further scientific efforts are still warranted to fully understand this deadly virus so that a cure can one day be discovered. As human and animal populations continue to grow, so does the cost and burden of this horrific disease. As a result, the importance of prophylaxis and passive immunity are critical in the event of medically managing an exposure, and preventing exposures. The World Health Organization has defined global recommendations for individuals and animals who have received prophylaxis to be adequately protected. Measuring this protection is performed using a variety of approved testing methodologies, virus-neutralizing assays and antigen-binding assays. Whilst the WHO recommendations were defined from clinical studies performed with virus-neutralizing assays, the assumption that these recommendations are suitable for the antigen-binding assays is inaccurate. The testing methodologies, virus-neutralization and antigen-binding, share similarities, as they are measuring an immune response to the rabies virus. However; enough differing characteristics are presented such that exact comparisons cannot be made. Establishing the same standards and recommendations for both testing methodologies will never be sufficient.

Master of Science / Department of Clinical Sciences / Maria S. Ferrer / Presence of anti-sperm antibodies (ASA) is associated with infertility in many species, including bulls but there is no standardized direct technique that allows detection of ASA bound to the sperm surface. The overall objective was to standardize a flow cytometric technique for detection of IgG and IgA directly attached to bovine sperm. The effects of fixation using phosphate buffer solution (PBS) or diluted formalin buffer solution (dFBS), exclusion of dead cells from the analysis, and aliquot variability were assessed using healthy bulls classified as Satisfactory Potential Breeders (SPB, n=9) and bulls with experimentally induced ASA (n=4) (Experiment1). The effect of freezing on the percentage of IgG- and IgA- bound sperm was assessed in samples from immunized bulls (n=4) (Experiment 2). Anti-sperm antibodies on the sperm surface were induced in yearling bulls by intramuscular injection of autologous semen and an adjuvant. Fixation of sperm cells did not affect the percentage of IgG- or IgA-bound sperm in any group of bulls. Exclusion of dead cell from the analysis did not affect the percentage of IgG-bound sperm (p= 0.0922 and p= 0.1525 for immunized and reproductively normal bulls, respectively). The exclusion of dead cells significantly increased the percentage of IgA-bound sperm in semen samples from immunized bulls (p= 0.0152) and significantly decreased the percentage of IgA- bound sperm in semen samples from reproductively normal bulls (p= 0.0012). Variability was < 10% in samples from immunized and reproductively normal bulls for percentage of IgG- and IgA-bound sperm. Freezing did not affect the percentage of IgG- (p=0.1287) or IgA-bound sperm (p=0.4175). Based on these results, fixation is neither necessary nor detrimental for analysis, and the percentage of antibody-bound cell should be calculated gated on the population of live cells only, especially when evaluating IgA binding. The percentage of ASA-bound sperm can be assessed on frozen-thawed samples. The development of this technique allows for further studies on ASA-bound sperm in populations of normal and abnormal bulls.

Master of Science / Department of Diagnostic Medicine/Pathobiology / Gary A. Anderson / Brucellosis is one of the major endemic zoonotic diseases worldwide, and it has history dating back to 1937 in Iraq when it was first isolated by an Iraqi physician. In order to establish a solution for the continuous devastating impacts of the disease in humans and livestock, the Brucellosis Control Program was established in 1995. The main responsibilities of this program were setting and implementing the appropriate strategies for controlling the disease. After the war in 2003, the United Nation organization for Food and Agriculture (FAO) developed a strategic plan to control the disease. The main goal of the project was to improve productivity in the livestock sector and reduce the prevalence of disease in small ruminants (sheep and goats) to less than 2%, and less than 0.2% in cattle and buffalo. Achieving such goals ultimately would reduce the disease incidence among the human population from more than 27.2 cases/100,000 persons in 2002, to less than 4 cases/100,000 people within 15 years. A serological surveillance was conducted and revealed the apparent prevalence of the disease in sheep and goats, cattle, buffalo, and camels was 6.51%, 1%, 1.48%, and 0.02%, respectively in Iraqi governorates except the three northern governorates of Kurdistan province . Based on surveillance results, a vaccination policy was the only appropriate strategy that could be chosen to control the disease. Four vaccination campaigns were implemented in 2006, 2007, 2008, and 2009, with a total number of vaccinated animals each year at 10099972, 4698482, 753153, and 1833482 head, respectively. The primary satisfactory outcome of the program was the apparent decline in livestock abortions leading to obvious increases in productivity. Regarding the incidence of brucellosis among the human population, the apparent decline in the middle and south of Iraq began with the vaccination phase of the control program in 2006. The results represented a significant decrease in human cases after only four vaccination campaigns of a program that was intended to continue for 15 years.

Master of Science / Department of Clinical Sciences / Rose M. McMurphy / Hypoventilation (PaCO2 > 45 mmHg) and large P(A-a)O2 gradients due to V/Q mismatch and shunt, are common during isoflurane anesthesia in horses. A fraction of inspired oxygen < 50% has been shown to improve ventilation and decrease intra-operative atelectasis in humans and some animals. The study compared the effects of two different fractions of inspired oxygen, 50% versus > 95%, on ventilation, respiratory pattern, and P(A-a)O2 gradient in isoflurane anesthetized horses. Eight mature horses were sedated with IV xylazine (1.0 mg/kg) and anesthetized with diazepam (0.05 mg/kg) and ketamine (2.2 mg/kg) twice. Anesthesia was maintained with isoflurane (ET1.5 vol%) in either 50 or > 95% oxygen for 90 minutes. Both treatments were randomly assigned to each horse with a one week interval in between treatments. Horses were positioned in dorsal recumbency, connected to a preloaded circle breathing system and allowed to spontaneously ventilate. Measurements included inspiratory and expiratory peak flow and time, tidal volume, respiratory frequency, ETCO2, CO2, O2, PaO2, PaCO2, pH, SaO2, heart rate, and arterial blood pressure. Calculated values included PAO2, P(A-a)O2, P(A-a)O2 rate of change, and physiologic dead space. FiO2 of 50% resulted in a lower PaO2, SaO2, PAO2, and P(A-a)O2. No significant change in PaCO2, ventilatory pattern, or any remaining measured variables was observed (p<0.05). The use of 50% oxygen and nitrogen as the carrier gas did not significantly change the ventilatory characteristics or improve oxygenation in isoflurane anesthetized horses. Repeatable respiratory rhythms characteristics were observed for horses while inspiring 50% and > 95% oxygen. A high A-a oxygen gradient with an equal rate of change overtime was still observed during both treatments.

Master of Science / Department of Clinical Sciences / Daniel Ulan Thomson / Consulting veterinarians (CV; n=23) representing 11,295,000 head of cattle on feed in the United States and Canada participated in a beef cattle health and well-being recommendation survey. Veterinarians were directed to an online survey to answer feeder cattle husbandry, health and preventative medicine recommendation questions. The CV visited their feedyards 1.7 times per month. All CV train employees on cattle handling and pen riding while only 13% of CV speak Spanish. All CV recommend IBR and BVD vaccination for high-risk (HR) calves at processing. Other vaccines were not recommended as frequently by CV. Autogenous bacterins were recommended by 39.1% CV for HR cattle. Metaphylaxis and feed-grade antibiotics were recommended by 95% and 52% of CV, respectively, for HR calves. Banding was more frequently recommended than surgical castration as calf body weight increased. The CV recommended starting HR calves in smaller pens (103 hd/pen) and allowing 13 inches/hd of bunk space. The CV indicated feedlots need to employ one feedlot doctor per 7,083 hd of HR calves and one pen rider per 2,739 hd of HR calves. Ancillary therapy for treating respiratory disease was recommended by 47.8% of CV. Vitamin C was recommended (30.4%) twice as often as any other ancillary therapy. Cattle health risk on arrival, weather patterns and labor availability were most important factors in predicting feedlot morbidity while metaphylactic antibiotic, therapy antibiotic and brand of vaccine were least important. This survey has provided valuable insight into feeder cattle health recommendations by CV and points to needed research areas.

Master of Science / Department of Diagnostic Medicine/Pathobiology / Weiping Zhang / Porcine epidemic diarrhea virus (PEDV) causes acute diarrhea to pigs at all ages, resulting in high mortality rate of 80-100% in piglets less than one week old. Within one year after the outbreak in April 2013, PEDV has rapidly spread in the US and causes the loss of over 10% of the US pig population. Monoclonal antibody (mAb) is a key reagent for rapid diagnosis of PEDV infection. In this study, we produced a panel of mAbs against nonstructural protein 8 (nsp8), spike(S) protein, and nucleocapsid (N) protein of PEDV. Four mAbs were selected, which can be used in various diagnostic assays, including indirect immunofluorescence assay (IFA), enzyme-linked immunoabsorbent assay (ELISA), Western Blot, immunoprecipitation (IP), immunohistochemistry (IHC) test and fluorescence in situ hybridization (FISH). The mAb 51-79 recognizes amino acid (aa) 33-60 of nsp8, mAb 70-100 recognizes aa1371-1377 of S2 protein, and mAb 66-155 recognizes aa 241-360 of N protein, while mAb 13-519 is conformational. Using the mAb70-100, the immunoprecipitated S2 fragment was examined by protein N-terminal sequencing, and cleavage sites between S1 and S2 was identified. In addition, this panel of mAbs was further applied to determine the infection site of PEDV in the pig intestine. IHC test result showed that PEDV mainly located at the mid jejunum, distal jejunum and ileum. Results from this study demonstrated that this panel of mAbs provides a useful tool for PEDV diagnostics and pathogenesis studies.

Le souffle équin est une maladie inflammatoire chronique des petites voies respiratoires, très fréquente chez les chevaux gardés à l’intérieur avec de la paille et du foin moisi et poussiéreux. Les signes cliniques peuvent être prévenus par le contrôle de l’environnement et soulagés par l’administration de corticostéroïdes systémiques et inhalés. L’objectif de cette étude était de déceler les effets secondaires présents sur des chevaux atteints de souffle traités à la fluticasone (Flovent 250 μg HFA®, 2000 μg BID, pendant six mois, et puis 2000 μg SID, pendant six autres mois) par le cortisol sérique et la présence d’ulcères gastriques. Cinq chevaux exempts de maladie respiratoire et onze chevaux atteints du souffle ont été gardés à l’intérieur d’une écurie avec du foin moisi et de la paille dans le but de provoquer la maladie chez le groupe atteints du souffle. Une fois les chevaux atteints de souffle devenus symptomatique, ils ont été divisés en deux groupes : un premier groupe traité avec de la fluticasone, nourri avec du foin et gardé sur une litière de paille, et un deuxième groupe non traité nourrie avec de la moulée et gardé sur une litière de ripe, pendant six mois. Par la suite, les deux groupes ont été mis au pâturage. Le cortisol a été mesuré par Immunoessai enzymatique par chimiluminescence (CEIA, Immunolite 1000, Siemmens®) les 12e et 10e jours avant et les 7e, 28e, 80e, 160e, 200e, 250e, 290e et 320e jours après le début du traitement afin de déterminer le degré de suppression du cortisol sérique. On a également fait une suivi de la présence d`ulcères gastriques à l`aide de vidéo endoscopique. La fluticasone inhalée deux fois par jour cause une diminution du cortisol sérique les 28e, 80e et 160e jours, mais elle n’entraîne pas d’effets sur le score des ulcères gastriques. Les pellets de luzerne causent quant à elles, une augmentation du score des ulcères gastrique chez les animaux exempts de maladie respiratoire. / Recurrent Airways Obstruction (RAO) is a small airways inflammatory disease, very common in horses stabled in mouldy-dusty hay and straw environments. The clinical signs are prevented by environmental control, relieved by systemic and inhaled corticosteroids. Our objectives were to determine whether inhaled corticosteroids cause a suppression of cortisol levels and gastric ulceration in RAO horses treated with fluticasone 2000μg (Flovant HFA®) BID for 6 months and 2000μg SID for another 6 months. Five (5) healthy horses were used as controls and eleven (11) RAO affected horses were stabled in a moldy-dusty environment to induce disease exacerbation. Once they were symptomatic, they were divided into two groups, the treated group was kept on hay/straw and the untreated group was fed with pellets food and bedded on wood shavings six months. Afterwards, all horses were pasture for the next 6 months. Serum cortisol was mesured by Immuno-essai enzymatique par chimiluminescence (CEIA, Immunolite 1000®, Siemmens) 12, 10 days before and 7, 28, 80, 160, 200, 250, 290, 320 days after treatment initiation, in order to determine cortisol suppression. Fluticasone administered twice a day reduces blood cortisol levels after 28, 80 and 160 days, but did not cause any change in gastric ulcers. However, pellets slightly increased gastric ulcer scores in healthy horses.

Master of Science / Department of Clinical Sciences / Walter Renberg / Objective: To evaluate the safety and effect of a single simultaneous intra-articular and intravenous injection of autologous adipose-derived stromal vascular fraction (SVF) and platelet rich plasma (PRP) on coxofemoral osteoarthritis (OA) in dogs. Methods: This was a randomized, double-blind, placebo-controlled prospective pilot trial of simultaneous intra-articular and intravenous SVF and PRP for coxofemoral OA. Dogs with coxofemoral OA causing signs of lameness or discomfort were evaluated by orthopedic exam, visual lameness score, Canine Brief Pain Inventory (CBPI), goniometry, visual analogue scale (VAS), and pressure-sensitive walkway (PSW) at week 0 (baseline), and at 4, 8, 12 and 24 weeks after injection. Joint radiographs were scored at 0 and 24 weeks. Results: Twenty two client-owned dogs with naturally occurring OA of the coxofemoral joints were enrolled (12 placebo-control, 10 SVF-treated). CBPI pain severity scores were lower in the treatment group at 24 weeks compared to the placebo group (p=0.042). The VAS score for the treatment group was significantly greater at 0 weeks than at 4, 8, or 24 weeks (p<0.05). When dogs with low quartile baseline PVF (25th percentile) were compared, the treatment group had statistically higher PVF at all post-injection time points when compared to the placebo group. After SVF injection, fewer dogs in the treated group were lame compared to the control group. Clinical Significance: This study is the first to utilize objective data from PSW as an outcome measure for dogs treated with SVF and PRP for coxofemoral OA. No adverse events were noted. Improvements in some measured parameters in the treated dogs compared to those in the placebo group.