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Master of Science / Department of Animal Sciences and Industry / Barry J. Bradford / The prevalence of fatty liver in transition dairy cattle has been reported to be as high as 50%. There are a few reliable on-farm diagnostic tools and even fewer methods to effectively prevent fatty liver. Non-alcoholic steatohepatitis, an advanced form of non-alcoholic fatty liver in humans, is accurately diagnosed with a commercial blood test that detects plasma cytokeratin-18 (CK18) fragments released during hepatocyte apoptosis. A study was performed using 89 Holstein cows in early lactation to determine if CK18 could serve as a novel indicator of liver triglyceride (TG) content. Although no previous work has been done with CK18 in bovine plasma, our results indicated that CK18 fragments were present in plasma. However, CK18 concentrations did not correlate with liver TG content or other measures of liver function, suggesting it is not a reliable diagnostic tool. Nevertheless, based on liver TG, plasma non-esterified fatty acid (NEFA), and plasma β-hydroxybutyric acid (BHBA) concentrations, this sample population as a whole was not suffering from severe metabolic problems or fatty liver, making it possible that plasma CK18 fragments are elevated only in the most extreme cases. Currently, there is no widely-adopted preventative strategy for fatty liver. A second study was performed to evaluate if encapsulated niacin (EN) could prevent liver TG accumulation during the transition period. Twenty-four primiparous (n=9) and multiparous (n=13) cows were randomly assigned to receive 0 or 24 g of dietary EN, beginning 3 weeks prior to expected calving until 21 days postpartum. Feeding EN did not influence liver TG content, but decreased plasma NEFA concentrations, suggesting inhibition of lipolysis. Multiparous EN cows also experienced depressed dry matter intake (DMI) in the 4 days prior to calving. However, even when EN reduced DMI, plasma NEFA was still suppressed. A novel finding was the prolonged clearance of caffeine in plasma on day 7 postpartum in EN-treated animals. In contrast to other studies, this dose and delivery method of EN did not result in an increase in plasma NEFA after EN treatment ended. These research projects determined that plasma CK18 is likely not a useful diagnostic tool for mild to moderate bovine fatty liver and that feeding EN can inhibit lipolysis but may influence DMI as well. This is one of the first studies into the metabolic effects of feeding EN, and further research is needed in this field.

Master of Science / Department of Clinical Sciences / Amy J. Rankin / Objective – To measure blood aqueous-barrier breakdown following aqueocentesis using various needle sizes and to monitor the intraocular pressure (IOP) response. Animals – 24 healthy, adult dogs received treatment (24 treated eyes, 24 contralateral eyes); 3 dogs were untreated controls (6 control eyes). Procedures – Dogs receiving treatment were divided into 3 equal groups (25-, 27-, or 30- gauge needle aqueocentesis). In each dog the treated eye was determined randomly, the contralateral eye was untreated. Dogs that did not have aqueocentesis performed in either eye were used as controls. Aqueocentesis at the lateral limbus was performed under sedation and topical anesthesia. Anterior chamber fluorophotometry was performed before and after aqueocentesis on day 1. On days 2-5 sedation and fluorophotometry were repeated. Intraocular pressure was measured with a rebound tonometer at multiple time points. Results – Aqueocentesis resulted in blood-aqueous barrier breakdown in all treated eyes with barrier reestablishment present by day 5 detected by fluorophotometry. On day 2 the contralateral untreated eyes of all groups also showed statistically significant increased fluorescence (P < 0.05) following treatment of the opposite eye, but these values were not statistically significantly greater than untreated controls. In treated eyes there was no statistical difference in fluorescein concentration or IOP between 27- and 30-gauge needles. Use of the 25- gauge needle resulted in a statistically significant increase in anterior chamber fluorescence on days 3 and 5. It also caused a statistically significant increase in IOP 20 minutes following aqueocentesis as compared to the 27- and 30-gauge needles. Aside from this transient ocular hypertension, rapid resolution of ocular hypotony following aqueocentesis was observed in all treatment groups. Conclusions and Clinical Relevance – Aqueocentesis using a 25-gauge needle resulted in a greater degree of blood-aqueous barrier breakdown and a brief state of intraocular hypertension following paracentesis. Use of a 27- or 30-gauge needle is recommended for aqueous paracentesis. A consensual ocular reaction appeared to occur in dogs following unilateral traumatic blood-aqueous barrier breakdown and may be of clinical significance. Statistical significance was limited in this study due to high variability and large standard deviations.

Doctor of Philosophy / Department of Diagnostic Medicine/Pathobiology / Daniel U. Thomson / Bovine viral diarrhea virus (BVDV) has emerged as one of the most important infectious diseases in cattle. One particular important manifestation, after successfully establishing an in utero infection of the fetus during the first trimester, is the development of a persistently-infected BVDV (PI-BVDV) calf. Persistently infected BVDV animals are a continuous source of virus and can shed the virus in virtually all secretions and excretions, including nasal discharges, saliva, semen, urine, tears, milk, and, to a lesser extent, feces. The objectives of this research were to determine: 1) the effects of short term exposure (13 – 18 days on feed (DOF)) to PI-BVDV feeder cattle; 2) the outcome of testing and removing PI-BVDV feeder calves at time of feedlot arrival on health, performance, and carcass characteristics; 3) the survival of BVDV on materials associated with livestock production; and 4) characterization of testing and longitudinal prevalences for PI-BVDV beef cattle. Testing and removing PI-BVDV calves at 13 to 18 DOF was too late to remove a morbidity effect due to PI-BVDV exposure. However, mortality, performance, and carcass characteristics were not different in cattle exposed to PI-BVDV cattle. Additionally, there were no harmful outcomes when newly arrived feeder cattle were exposed to a PI-BVDV animal for one to two days following feedlot entry. A non-cytopathic, Type 1b, BVDV was capable of surviving after application to various materials used in livestock production. BVDV tended to survive longer on non-porous materials than porous materials. When in the presence of mucus, BVDV was protected from degradation for longer periods of time than when not in the presence of mucus. There was no difference in overall PI-BVDV prevalence within cattle sampled in 2006 and 2007. Cattle that weighed less than 300 lbs. had a greater likelihood of being PI-positive than cattle with increased weights. Several months of the year had a greater likelihood of having PI-positive animals. Based on operation, cow-calf and stocker operations had a greater likelihood of having PI-positive animals than did feedlot operations.

La leptospirose est une zoonose à distribution mondiale dont la prévalence chez le chat varie géographiquement de 4.8% à 35%. Bien que l’exposition féline à Leptospira spp. soit rapportée dans des études sérologiques, les conséquences cliniques de cette maladie chez le chat sont peu connues. Le but principal de cette étude était de comparer le statut sérologique et de porteur (PCR urinaire) de Leptospira spp. entre des chats sains et des chats atteints de maladie rénale (insulte rénale aigue et maladie rénale chronique de stades IIb, III et IV). Une étude préliminaire pour valider la sensibilité et la spécificité analytiques de la PCR de Leptospira spp. réalisée par le Laboratoire de Diagnostic Moléculaire de la FMV sur l’urine de chat a été effectuée. La validation in vitro a démontré que la technique de PCR est efficace pour déterminer la présence de leptospires pathogènes dans l’urine du chat. Dans le cadre de l’étude principale, 251 chats ont été recrutés entre janvier 2010 et mars 2012,. De ceux-ci, 240 ont été inclus et divisés en 2 groupes (chats sains (C=125) et chats atteints de maladie rénale (MR=115) en se basant sur un examen physique ainsi que sur des résultats d’hématologie, de biochimie et d’analyse d’urine. Tous les chats recrutés ont également été examinés sérologiquement par test de micro-agglutination pour la présence d’anticorps contre Leptospira spp. (résultat considéré positif si ≥1 :100) et par PCR pour la présence de Leptospira spp. dans l’urine. Le pourcentage prédit de séropositivité pour Leptospira spp. était significativement plus élevé chez les chats atteints de maladie rénale (13,7%) que chez les chats sains (5%) (p=0,02). Les sérovars impliqués étaient Pomona (n=16), Bratislava (n=8) et Grippotyphosa (n=1). De plus, les chats séropositifs pour Pomona présentaient des titres significativement plus élevés que pour les autres sérovars (p=0,04). L’excrétion de Leptospira spp. a été confirmée par PCR dans l’urine de huit chats. Des 26 chats séropositifs, quatre (C=2, MR=2) se sont également révélés PCR positifs. La prévalence a été plus élevée chez les chats du groupe MR (5.3%; 6/113) lorsque comparée à celle du groupe C (1.6%; 2/125), mais cette différence ne s’est pas révélée statistiquement significative (C=0,9% , MR= 5,5% ; p = 0,09). L’âge, le sexe et le milieu de vie (urbain versus rural) n’ont pas influencé le statut sérologique ou d’excrétion pour Leptospira spp. Le pourcentage prédit de séropositivité était significativement plus élevée chez les chasseurs (p<0.01) et pendant les mois de juin à août (p=0.02). La présence d’un autre chat à la maison a également significativement augmenté ce pourcentage (p<0.01), mais la présence d’un chien ne l’a pas influencé. Lors de l’évaluation du PCR par le modèle GGE, seules les variables « contact avec raton laveur » et « contact avec mouffettes » sont ressorties statistiquement significatives (p≤0.03). Le rôle que joue Leptospira spp. comme agent étiologique de maladie rénale chez le chat demeure incertain. Toutefois, la différence significative de statut sérologique entre les chats sains et les chats atteints de maladie rénale suggère que la leptospirose pourrait être une cause sous-diagnostiquée de maladie rénale chez cette espèce. Dans cette étude, plusieurs porteurs asymptomatiques ont été identifiés, ce qui suggère que l’espèce féline puisse être un acteur sous-estimé dans la transmission de la bactérie aux humains. / Leptospirosis is a globally widespread zoonosis, with prevalence in cats varying from 8.8% to 35% depending on geographical localization. Although serologic evidence of feline exposure exists, clinical disease is rarely reported. This study aimed to compare seropositivity and urinary PCR status for Leptospira spp. between healthy cats (H) and cats with kidney disease (KD: acute kidney injury (AKI) and chronic KD stages IIb, III and IV). The analytical sensitivity and specificity of the PCR performed by the Laboratoire de Diagnostic Moléculaire of the FMV were evaluated. In vitro validation showed that the PCR technique is effective for determining the presence of pathogenic leptospires in the urine of cats. A total of 251 cats were recruited, from which 240 cats were enrolled. Cats were enrolled from January 2010 to March 2012 and divided into two groups, H (n=125) and KD (n=115), based on complete blood count, serum biochemistry profile and urinalysis. Leptospira spp. serology by microscopic agglutination test (titers ≥ 1:100 considered positive) and urinary PCR were performed in all cats. Predicted seropositivity for Leptospira spp. was statistically different between groups; being 5% and 13,7% in the H and KD groups respectively (p=0.02). Serovars involved were Pomona (n=16), Bratislava (n=8) and Grippotyphosa (n=1), with titers being significantly higher for Pomona (p=0.04). The excretory status was confirmed by a positive urine PCR in eight cats. Of the 26 seropositive cats, four (H=2, KD=2) were also PCR positive. The prevalence of PCR positive cats was higher in the KD group (5.3%; 6/113) compared with the H group (1.6%; 2/125), but the difference between groups did not reach statistical significance (0.9% in H, 5.5% in KD; p=0.09). Age, sex and rural versus urban environment did not influence serologic or PCR status for Leptospira spp. Predicted seropositivity was greater between June and August (p =0.02) and in known hunters (p<0.01). The presence of another cat at home also increased significantly the predicted seropositivity (p<0.01), although the presence of a dog did not. When evaluating the PCR status of cats in the GEE model for individually tested variables, only the variables “contact with raccoons” and “contact with skunks” were statistically significant (P ≤ .03). Although the precise role of Leptospira spp. as an etiologic agent of feline KD remains unclear, the significant difference in the serologic status found between H and KD cats suggests that it may be an under-diagnosed cause of KD in cats. Several asymptomatic carriers were identified, suggesting that cats could be underestimated players in the transmission of the bacteria to humans.