Bien que les infections à Besnoitia tarandi sont documentées chez l’espèce Rangifer sp. depuis 1922, les données récoltées sur l’écologie et la distribution de cette parasitose demeurent rares. Les objectifs de cette étude ont donc été (i) d’identifier le meilleur tissu à échantillonner pour détecter les infections à Besnoitia tarandi dans les populations de caribous, (ii) de calculer la sensibilité et la spécificité de l’examen visuel comparativement à l’examen microscopique et (iii) d’identifier les facteurs de risques intrinsèques et extrinsèques associés à cette parasitose afin (iv) de comparer la prévalence et la densité des kystes parasitaires entre certains troupeaux. Nos résultats suggèrent que l'examen microscopique du derme superficiel d’une section de peau provenant du tiers moyen antérieur du métatarse devrait être privilégié pour dépister les infections par B. tarandi et en évaluer l'intensité. L’examen microscopique est également un outil très sensible comparativement à l’examen visuel des kystes parasitaires. Besnoitia tarandi, qui semble être absent du Groenland, a été observé dans environ un tiers des caribous nord-américains. Une variation saisonnière de prévalence et d'intensité de B. tarandi a été détectée; le parasite étant plus abondant chez cet hôte intermédiaire durant la période de l'automne/hiver comparativement à celle du printemps/été. Cet effet saisonnier pourrait être associé à une augmentation de l'abondance du parasite suite à la saison des insectes (i.e. été), supportant ainsi le rôle présumé des arthropodes piqueurs comme vecteurs de la maladie. Cette différence saisonnière pourrait aussi être expliquée par la diminution de la charge parasitaire par le système immunitaire et/ou par un taux de survie inférieur des animaux les plus parasités durant la saison froide. Les niveaux d'infection étaient légèrement plus élevés chez les mâles que chez les femelles, ce qui suggère soit une diminution du taux de mortalité, soit une exposition accrue ou une plus grande susceptibilité au parasite des mâles en comparaison aux femelles. La densité d’infection supérieure dans le troupeau Rivière-aux-Feuilles (Nunavik) suggère des niveaux d'exposition au parasite plus élevés et/ou une diminution des niveaux de résistance de ces caribous à ce protozoaire. Les résultats de cette étude démontrent que B. tarandi peut réduire les chances de survie des caribous infectés. Il sera donc important de continuer à surveiller les infections à B. tarandi surtout en cette période de changements climatiques. / Although Besnoitia tarandi has been described in Rangifer sp. for over 80 years, understanding of its ecology and distribution is still relatively limited. The objectives of this study were (i) to determine the anatomical sampling site that enhances detection of Besnoitia tarandi infections in caribou populations; (ii) to evaluate the relative sensitivity and specificity of the in situ macroscopic assessment for diagnosis of besnoitiosis compared to the microscopic evaluation; and (iii) to identify potential risks factors associated with the prevalence and intensity of B. tarandi cysts and compare these across different populations. Our results suggest that calculating the number of cysts present in the superficial dermis of a skin section of the anterior mid-third of the metatarsus region by microscopic examination should be favored in order to monitor the presence and intensity of B. tarandi infections. Macroscopic assessment of B. tarandi cysts was not a sensitive method compared to that of a microscopic analysis. Besnoitia tarandi seems to be absent from Greenland but has been encountered in approximately one third of North-American caribou. A seasonal effect was noticed in the prevalence and intensity of B. tarandi infections; the parasite being more abundant in caribou sampled during the fall/winter period compared to the spring/summer period. This effect could reflect the increase abundance of B. tarandi following the end of the insect season (i.e. summer), supporting the role of arthropods as vectors of transmission. Reduction of the parasite burden by the immune system and/or a lower winter survival rate of severely infected caribou could explain the seasonal difference. The slightly higher prevalence in males suggests lower mortality rate, higher exposure and/or higher susceptibility to the parasite in infected males when compared to infected females. The apparent higher density of infection by B. tarandi in the Rivière-aux-Feuilles herd (Nunavik, Québec) suggests either higher exposure to the parasite or reduced resistance of caribou from this herd. The findings of this study suggest that infection by B. tarandi might reduce survival of individual animals; hence, the dynamic between this parasite and its hosts is worth further investigation, especially in these days of changing Arctic environment.

Peu d’études ont exploré l’utilisation du traitement prolongé pour les mammites cliniques et aucune ne portait sur l’utilisation du traitement prolongé à base de ceftiofur pour le traitement des mammites cliniques légères à modérées. L’objectif de cette étude était d’évaluer l’efficacité du traitement intra-mammaire prolongé à base de ceftiofur pour les mammites cliniques légères à modérées, en considérant toutes les bactéries responsables ou seulement Staphylococcus aureus (S. aureus) et les streptocoques. Des vaches laitières (n = 241) de 22 élevages du Québec et de l’Ontario ont été inclues. L’étude a été conçue comme un essai clinique à allocation aléatoire. Pour chaque cas de mammite clinique légère à modérée, 125 mg d’hydrochloride de ceftiofur (Spectramast® LC) a été administré par voie intra-mammaire une fois par jour pour 2 ou 8 jours. Le pourcentage de guérison clinique 21 jours après la fin du traitement était de 89% (n = 98/110 pour chaque groupe, p = 0,95). Les pourcentages de guérison bactériologique 21 jours après la fin du traitement pour les groupes 2 jours et 8 jours étaient 32% (n = 15/47) et 61% (n = 25/41) respectivement pour toutes les bactéries (p < 0,01); 64% (n = 9/14) et 82% (n = 9/11) respectivement pour les streptocoques (p = 0,50); et 0% (n = 0/20) et 47% (n = 9/19) pour S. aureus (p < 0,01). Il n'y avait pas de différence entre les 2 groupes pour les nouvelles infections intra-mammaires (p = 0,30). Le traitement prolongé à base de ceftiofur est un choix raisonnable pour le traitement des mammites cliniques légères à modérées, en particulier causées par S. aureus. / Little research has focused on extended therapy in lactating cows with clinical mastitis and none were with ceftiofur. The objective of the present study was to evaluate the efficacy of an intramammary extended ceftiofur treatment for mild to moderate clinical mastitis, and to determine whether it would increase cure rates when considering any bacteria or more particularly Staphylococcus aureus (S. aureus) and streptococci. Holstein dairy cows (n = 241) from 22 herds located in Quebec and Ontario, Canada, were included. The study was designed as a randomized clinical trial. For each case of mild to moderate clinical mastitis diagnosed, 125 mg of ceftiofur hydrochloride (Spectramast® LC) was administered intramammary once a day for 2 or 8 consecutive days. Clinical cure rate 21 days after the last treatment was identical in each group (89%; n = 98/110; p = 0.95). Bacteriological cure rates 21 days after the last treatment for the 2- and 8-day regimens were 32% (n = 15/47) and 61% (n = 25/41), respectively, for all bacteria (p < 0.01); 64% (n = 9/14) and 82% (n = 9/11), respectively, for streptococci (p = 0.50); and 0% (n = 0/20) and 47% (n = 9/19), respectively, for S. aureus (p < 0.01). There were no statistical differences between experimental groups for new intramammary infections. Extended therapy appears to be a reasonable choice for the treatment of mild to moderate clinical mastitis, particularly in cases where S. aureus is the pathogen involved.

L’implantation retardée ou diapause embryonnaire décrit l'arrêt ou le retardement pendant l'embryogenèse. Chez le vison, la diapause est corrélée avec une sécrétion pituitaire insuffisante de la prolactine, ayant pour résultat la différentiation incomplète du corpus luteum et réduction de la progestérone. Des études antérieures suggèrent que le blastocyste de vison en diapause demeure dans un état de quiescence ou se développe lentement. Pour élucider ceci, la réplication de l'ADN a été étudiée. Les résultats démontrent synthèse de l’ADN et prolifération cellulaire dans les embryons au stade de morula, avant la diapause et dans les blastocystes après la réactivation. La réplication de l'ADN a été également détectée dans des blastocystes en diapause et en diapause prolongée. L'implantation est considérée comme une interaction bidirectionnelle entre le blastocyste et l'utérus. Il a été montré que les prostaglandines sont importantes pour la vascularisation de l’utérus au moment de l’implantation et peuvent réactiver l'utérus des visons après la diapause. La concentration protéinique et la localisation de la phospholipase citosolique A2 (CPLA2) et de la cyclooxygenase 2 (COX2) ont été étudiées dans l'utérus de vison. L’expression de la CPLA2 et COX2 étaient sur-régulées au moment de l'implantation. Il est connu que la prolactine active les corpus luteum des visons. L'idée de un lien entre la prolactine et la voie de signalisation des prostaglandines a été testée en mesurant les récepteurs de prolactine. Les résultats montrent une augmentation de l’expression des récepteurs de prolactine à l'implantation suggérant que la prolactine pourrait activer la voie de prostaglandine à l'utérus par son propre récepteur. La conclusion, les embryons pendant la diapause ne sont pas arrêtées complètement et les protéines liées à la voie de prostaglandine sont implique dans la réactivation de l'utérus. / Delayed implantation or diapause describes arrest or retardation during embryogenesis. In mink, diapause is related to insufficient pituitary prolactin secretion, resulting in incomplete differentiation of the corpus luteum with reduced progesterone concentration. The mink blastocyst at diapause was believed to be totally quiescent or expanding at a low rate. To explore this, DNA replication was studied. Results showed synthesis of DNA, and thus cell proliferation at the morulae stage before diapause and at the blastocyst following activation. DNA replication was detected not only at diapause but also at extended diapause. Furthermore, implantation is considered as a two-way interaction between the blastocyst and the uterus. It has been shown that prostaglandins are important for vascularization of the uterus and products of the prostaglandin pathway could reactivate the mink uterus following diapause. Protein concentration and localization was studied for cytosolic phospholipase A2 (CPLA2) and cyclooxygenase 2 (COX2) in mink uterus. Expression of CPLA2 and COX2 was up regulated at implantation. It is know that prolactin is the factor that activates the mink corpus luteum. The idea of a link between prolactin and prostaglandin pathway was investigated by quantifying the prolactin receptors in the uterus. Results showed an increase of prolactin receptors at implantation suggesting that prolactin could activate the prostaglandin pathway at the uterus through its own receptor. In conclusion, embryos during diapause are not completely arrested, and proteins related to the prostaglandin pathway are implicated in reactivation of the uterus.

Au cours des dernières années, Salmonella Enteritidis est devenus les sérotypes les plus souvent isolés chez les patients canadiens, les cas étant liés à la consommation de viande de poulet et d’œufs crus. Les vaccins tués commercialement disponibles pour la volaille, stimulent mal l'immunité mucosale, tandis que l'utilisation de vaccins vivants reste controversée. Par conséquent, un vaccin sous-unitaire par voie orale peut être une solution. Cinq protéines bactériennes ont été choisies comme candidates potentielles et identifiées, soit Glyceraldehyde-3-phosphate dehydrogenase, Enolase, Lipoamide dehydrogenase, DNA protection during starvation protein et Elongation factor-Tu. Notre objectif a été de produire et de purifier ces protéines et de démontrer leur immunogénicité. Les gènes des protéines ont été amplifiés et clonés dans le vecteur pQE-30 pour expression dans Escherichia coli M15. La purification a été effectuée par FPLC. Des poules pondeuses SPF ont été séparées en 6 groupes et injectées par voie intramusculaire à different âges avec une des 5 protéines, ou le PBS chez le groupe témoin. Les œufs ont été ramassés pendant l'expérience et du sang a été prélevé à 36 semaines d'âge. Les anticorps IgY ont été extraits à partir du jaune d'oeuf et du sérum, et les IgA à partir du blanc d'oeuf. Des immunodots, westernblots et ELISA ont évalué l'immunogénicité des protéines et les niveaux d'anticorps induits . Nous avons constaté que ces cinq protéines pourraient stimuler la production d'anticorps spécifiques in vivo. GAPDH, Enolase et DPS ont induit des titres d'anticorps plus élevés que LpdA et EF-Tu. / Over the past years, Salmonella Enteritidis (SE) has become the most prevalent serovars isolated in Canadian patients. Most cases in humans are associated with consumption of chicken meat, raw egg and related products. For controlling Salmonella transmission and infection in poultry, available commercially killed vaccines poorly stimulate mucosal immunity, while the use of live vaccines remains controversial. Therefore an oral subunit vaccine may be a solution. Five bacterial proteins were chosen as potential candidates and identified as Glyceraldehyde-3-phosphate dehydrogenase, Enolase, Lipoamide dehydrogenase, DNA protection during starvation protein and Elongation factor-Tu. Our objectives were to produce and purify these proteins and study their immunogenicity. The proteins genes were amplified and cloned into pQE-30 vector, then transformed into Escherichia coli M15 for expression. Purification was performed using FPLC. SPF laying hens were separated into 6 groups and injected intramuscularly 3 times at 16, 20 and 28 weeks of age. Five groups were injected with a single protein respectively while the sixth group was injected with PBS as control. Eggs were collected during the duration of the experiment and blood was collected when hens were sacrificed at 36 weeks of age. IgY was extracted from egg yolk and serum and IgA from egg white. Immunodot, westernblot and ELISA were used to evaluate the immunogenicity of proteins and antibody levels they induced. We found that these five proteins could stimulate production of specific antibody in vivo. GAPDH, Enolase and DPS induced higher antibody titer than LpdA and Ef-Tu.

Mycobacterium avium ssp. paratuberculosis (MAP) est l'agent causal de la paratuberculose, maladie entérique, chronique et incurable des ruminants, avec un impact économique important. Une meilleure compréhension des facteurs de risque associés à l'introduction de la maladie dans un troupeau est essentielle pour sa prévention. L’amélioration des tests diagnostiques est aussi importante pour son contrôle. L’introduction des nouveaux animaux dans le troupeau et la présence et contact des différentes espèces sauvages et domestiques avec les vaches, semblent être des facteurs de risques d’introduction de la maladie. Nous avons réalisé une revue systématique dont l`objective était de recueillir l’information pertinente pour répondre à la question sur l’importance de ces facteurs et leur impact sur l’introduction de la maladie dans un troupeau. D`un autre côté, la détection de MAP dans les fèces par culture bactérienne demeure la méthode diagnostique de choix malgré les facteurs qui l`affectent. Une série de 3 étapes est requise afin de confirmer la présence du MAP : (1) culture (2) coloration, et (3) confirmation du MAP par PCR (si détecté à l´étape 2). Certains échantillons fécaux présentent une particularité en raison de leur forte charge de micro-organismes. Ces contaminants peuvent interférer avec la croissance et la détection de MAP. Une étude visant à : a) estimer l'impact des certain covariables sur les résultats de la culture de MAP parmi l`analyse rétrospective d`un banque des données et b) évaluer la possibilité d'optimiser le processus de diagnostic du MAP en effectuant l'analyse PCR sur les cultures déclarées comme contaminées a été réalisée. / Mycobacterium avium ssp. paratuberculosis (MAP) is the causative agent of paratuberculosis an enteric, chronic and incurable disease of ruminants, with a major economic impact. A better understanding of the risk factors associated with the introduction of the disease in a herd, is essential for its prevention. Improving diagnostic tests are also important for the control. The introduction of new animals into the herd and the presence and contact of wild and domestic species with cattle, appear to be a risk factor for disease introduction. We conducted a systematic review to obtain relevant information to answer the question about the importance and impact of these factors on the introduction of the disease into a herd. On the other hand, the detection of MAP in feces by bacterial culture remains the diagnostic method of choice despite of the factors that can affect it. A series of 3 steps are required to confirm the presence of MAP: (1) culture (2) acid fast stain (3) MAP confirmation by PCR (if detected in step 2). Some fecal samples exhibit a particularity due to their normal heavy load of micro-organisms. These contaminants can interfere with the growth and detection of MAP. A study to: a) assess the impact of some covariables on the culture results by a retrospective analysis of a databank and b) to evaluate the possibility to optimize MAP diagnostic process by performing a PCR analysis on cultures declared as contaminated was realized.

No description available.

Master of Science / Department of Clinical Sciences / Thomas Schermerhorn / Tonicity is property of a solution that is defined as the total effective (impermeable) osmole concentration that drives fluid movement across a semipermeable membrane via osmosis. Tonicity is related to but distinct from solution osmolality, which is a summation of all solute concentrations, regardless of the solute membrane permeability. In the mammalian body, tonicity is tightly regulated at both a cellular and systemic level; tonic derangements cause rapid change in cell and tissue volume leading to significant dysfunction. Input from the central nervous, circulatory, endocrine, gastrointestinal, and urinary systems are integral to osmoregulation, so many diseases in veterinary medicine are associated with tonicity disorders. However, because the homeostatic mechanisms that control tonicity overlap with those regulating electrolyte and acid-base balance as well as hydration and vascular volume, tonic consequences of disease can be difficult to isolate. Understanding of disease-associated changes in tonicity is further complicated by the fact that the tonic contributions of many solutes that accumulate in disease are unknown. Additionally, direct assessment of tonicity is difficult because tonicity is not just a physiochemical property, but it implies a physiologic effect. Thus, simple summation of osmole concentrations is an inadequate measurement of tonicity. The following report includes three studies investigating various aspects of tonicity as it applies to veterinary patients. Chapter 2 reports a study that examines the tonic effects of ketoacids and lactate using two different in vitro red blood cell assays. Results demonstrated that the ketoacids, beta-hydroxybutyrate and acetoacetate, behave as ineffective osmoles while the tonic behavior of lactate is variable, implying a more complex cellular handling of this anion. Two additional studies examine whether the mean corpuscular volume difference (dMCV) is a novel clinical marker for hypertonicity in dogs. Results of separate retrospective (Chapter 3) and prospective (Chapter 4) studies provide evidence that dMCV is a useful clinical marker for hypertonicity in dogs.

No description available.

Doctor of Philosophy / Department of Anatomy and Physiology / Bruce D. Schultz / The goal of this project was to define mechanisms responsible for Na+ transport in two hormonally-sensitive epithelium, the bovine mammary gland and porcine vas deferens. Glucocorticoid stimulation in these epithelia results in a significant increase in amiloride-sensitive ion transport, suggesting regulation of the epithelial Na+ channel, ENaC. ENaC has typically been described as a heteromultimeric ion channel with at least three different types of subunits, the most common being , β, and γ. Glucocorticoid-induced regulation of these subunits at the transcriptional level appears to be very different in the porcine vas deferens as compared to the bovine mammary gland. The aims of the study in mammary epithelium were to elucidate the mechanisms by which apical electrolytes and cytokines compromise barrier function in mammary epithelium. The long term goal is to better understand and manage the interaction between ionic composition of milk and breakdown of the gland epithelium that occurs during mastitis. Our results suggest a causal link between changes in milk electrical conductivity and epithelial barrier breakdown that has not been appreciated previously. Results will provide benefits to dairy farmers by characterizing steps that might prevent the development of mastitis or hasten recovery. The aims of the study using porcine vas deferens epithelial cells include determining the time course, concentration- and structure-dependency for regulation of amiloride-sensitive ion flux by corticosteroids. Corticosteroids caused a concentration-dependent increase in amiloride-sensitive Isc with a rank order of potency of dexamethasone>prednisolone>cortisol. Hill analysis indicates steep concentration dependency. The corticosteroid-induced, amiloride-sensitive current is Na+ absorption as indicated by radiotracer flux measurements. Studies employing selective antagonists (spironolactone, mifepristone) define glucocorticoid receptor mediation. These results suggest that vas deferens epithelia are exquisitely sensitive to corticosteroid exposure. Observed changes in epithelial function in response to corticosteroid exposure would rapidly and chronically affect the luminal environment to which sperm are exposed. Thus, physiological and pharmacological corticosteroid exposure is expected to affect male fertility.

Doctor of Philosophy / Department of Diagnostic Medicine/Pathobiology / Tiruvoor G. Nagaraja / George C. Stewart / Fusobacterium necrophorum, a gram negative anaerobe and an opportunistic pathogen, causes necrotic infections in humans and animals. Two subspecies of F. necrophorum, subsp. necrophorum and subsp. funduliforme are described. Leukotoxin (Lkt), a secreted protein encoded by a tricistronic operon (lktBAC), is the major virulence factor of F. necrophorum. The concentration of Lkt produced by subsp. necrophorum is higher than that of subsp. funduliforme. Quantitative-PCR was used to determine the relative expression of lktA by the two subspecies of bovine origin. The mRNA transcript of lktA was detectable in early-log phase of growth in subsp. necrophorum, whereas in subsp. funduliforme, the lktA transcript was detected only in the mid-log phase. Q-PCR analysis revealed that subsp. necrophorum had 20-fold more lktA transcript than subsp. funduliforme. The amount of lktA transcript declined by late-log phase in both subspecies; but lktA mRNA levels in subsp. necrophorum was 8-fold higher than in subsp. funduliforme. Leukotoxin protein stability assays showed the Lkt to be stable in both subspecies despite the decrease in the concentration of the protein during late-log phase. The subspecies identity of human F. necrophorum strains and whether they possess lktA and leukotoxin activity are not known. Human clinical isolates (n = 4) of F. necrophorum were identified as subsp. funduliforme based on 16S rRNA sequence and absence of hemagglutinin gene. Four human strains had the lkt promoter, lktB, and lktC similar to that of subsp. funduliforme. One strain had full length lktA, while other three strains exhibited considerable heterogeneity. All four strains secreted Lkt that was toxic to human leukocytes. Fusobacterium equinum, formerly F. necrophorum, is a newly recognized species. It is associated with infections of the respiratory tract in horses. Little is known about the virulence factors of F. equinum. Southern hybridization revealed that F. equinum strains had lktA gene with greater similarities to F. necrophorum subsp. necrophorum. The toxicity of culture supernatants of isolates to equine leukocytes was variable. Our data indicate that F. equinum isolates possess lktA gene and exhibit leukotoxin activity. The importance of leukotoxin as a virulence factor in human and equine fusobacterial infections needs to be investigated.