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Loss of adaptive variation arising from population declines and fragmentation is a primary concern in conservation. However, many conservation programmes assess only neutral genetic variation. Whilst assessments of neutral variation are informative about demographic history, inbreeding and genetic structure, they do not provide information on adaptive variation. The Major Histocompatibility Complex (MHC) is a group of genes that has been extensively studied and are known to be important in effective immune responses. Given the threat posed by infectious diseases to wildlife, the MHC is increasingly being assessed in endangered species. African wild dogs (Lycaon pictus, hereafter wild dog) are an endangered canid that has suffered extensive declines in the wild and now persist as small and fragmented populations totalling less than 8,000 individuals. The purpose of this study was to assess how neutral and MHC marker data genetic data can be used to assist conservation of this species. As such, I assessed sequence diversity across ~300bp of mitochondrial DNA, patterns of polymorphism and heterozygosity at 10 neutral microsatellite loci, compared to sequence variation and haplotype diversity at the MHC. Wild dogs were found to be genetically depauperate at the MHC compared to other canids. Patterns of variation indicate a historical loss of variation, followed by more recent diversification. However, it was also shown that evolutionary history contributes to differences in diversity between species. The spatial and temporal structure of MHC diversity was found to be largely correlated with neutral markers, which may suggest that selection is unable to counter strong genetic drift in such small populations. Overall, genetic diversity of both neutral and MHC markers appeared to be largely determined by demographic stability and size of populations. Habitat fragmentation and loss were associated with genetic isolation of wild dog populations, which showed strong structuring. However, the barriers to, or corridors for, dispersal of wild dogs were not always clear. The European captive breeding population was found to have comparable diversity metrics to wild populations, and was found to contain a large proportion of the MHC variation from the Southern African populations from which they were originally sourced. Careful genetic management is now required to correct the severe over- and underrepresentation of some founder lineages in this captive population to reduce inbreeding and loss of genetic variation.

In vitro maturation of dog oocytes has always been the main obstacle preventing reproductive biologist from producing canine in vitro cultured embryos. The unsuccessful oocyte maturation in canine species originates from their unique physiological and biological specifications. Ovulation of dominant follicles in bitch (6-12 in each oestrous cycle) occurs at prophase I stage of oocyte nucleus and meiotic resumption develops during 3-5 days of oviductal transition. During this PhD thesis, studies were designed in order to speculate characteristics of canine oocyte maturation in vitro in terms of maturation media components, gas composition of the incubator and hormonal requirements. Level of oxidative stress during 72h (culture period) of in vitro maturation showed that 5%O2, 5% CO2 and 90% N2 composition improves meiotic resumption and reduces degeneration rate significantly compared to 5% CO2 in air. Utilization of caffeine as a non specific phosphodiesterase inhibitor at 10mM for 12h at the beginning of the 72h culture (12+60) also improved MII maturation rate (16.9% ± 2.4; P < 0.05). Among several hormonal treatments recombinant porcine Growth Hormone (PGH) at 100ng/ml and Melatonin (MTN) at 100nM concentrations had outstanding improvement over meiotic resumption (28.9% ±10.0 and 56.2% ±8.6 respectively; P < 0.05). Attempts were made to study developmental potentials of optimally matured oocytes by parthenogenetic activation (PA) and in vitro fertilization (IVF) using chilled semen. Partial digestion of the zona pellucida prior to IVF improved the cleavage rate at 48h 6.4% ± 0.3 and resulted in production of a single 8 cell embryo. Moreover; canine follicular cells were culture in order to characterize their primary culture morphology and steroidogenic responsiveness to physiological and pharmaceutical substances. Immunolocalization of aromatase (CYP19) positive cell clumps, presumptive oestrogen producing colonies, was identified. This primary culture also maintained its steroidogenic machinery up to 96h (measured by radioimmunoassay) with a significant increase in production of estradiol and progesterone after 72h compare to the start of the culture.

Objectifs : valider des équations prédictives de la teneur en eau totale (TBW) chez le chien vigile de race beagle par bioimpédancemétrie monofréquence à 50 kHz, en comparaison avec la méthode de référence de dilution au deutérium. Ces équations seront ensuite appliquées à différentes races de chiens afin de vérifier leur validité chez tous les formats de chiens.Animaux : 26 chiens de laboratoire de race beagle et 13 chiens de propriétaires de diverses races. Méthodes : TBW est déterminée par la méthode de référence de dilution à l’eau deutérée et indirectement par l’utilisation d’équations prédictives établies à partir de mesures morphologiques et des mesures électriques (résistance et réactance) obtenues au moyen d’un bioimpédancemètre. Résultats : Les données obtenues chez 16 des 26 beagle a permis d’établir, par régression linéaire, deux équations prédictives de la teneur en eau totale suivantes : TBW1 = –0.019(BL2/R) + –0.199(RC + AC) + 0.996W + 0.081H + 12.31; et TBW2 = 0.048(BL2/R) + –0.144(RC +AC) + 0.777W + 0.066H + 0.031X + 7.47, où AC est le périmètre abdominal, H la hauteur au garrot, BL la longueur du corps, R la résistance, RC le périmètre thoracique , W le poids; and X la réactance. Chez les 10 beagles restants, TBW calculée par les équations TBW1 et TBW2 et celle obtenue par la méthode de dilution sont fortement corrélées (R21 = 0.843 ; R22 = 0.816). La 1ère et la 2ème équations sous-estiment de façon non significative la TBW de respectivement 2.4% and 1.6%. Cependant, l’application de ces formules à des chiens de diverses races ne permet pas l’estimation correcte de la TBW par rapport à la méthode de référence.Conclusion : Cette étude montre qu’il est possible et facile d’utiliser la bioimpédance chez le chien vigile. Deux équations prédictives de la TBW ont été développées et validées chez le beagle mais ne sont pas applicables en l’état à d’autres races de chien. La diversité morphologique des races canines obligent à adapter ces équations en fonction de paramètres de conformation et de format. / Objectives—To develop equations for prediction of total body water (TBW) content in unsedated dogs by combining impedance and morphological variables, and to compare the results of those equations with TBW content determined by deuterium dilution (TBWd). Then to investigate whether these equations were predictive of TBW in various canine breeds.Animals—26 healthy laboratory adult Beagles and 13 healthy adult pet dogs of various breeds.Procedures—TBW content was determined directly by deuterium dilution and indirectly with equations developed from measurements obtained by use of a portable bioelectric impedance device and morphological variables.Results—Impedance and morphological data from 16 of the 26 Beagle dogs were used to determine coefficients for the following 2 equations: TBW1 = –0.019(BL2/R) + –0.199(RC + AC) + 0.996W + 0.081H + 12.31; and TBW2 = 0.048(BL2/R) + –0.144(RC +AC) + 0.777W + 0.066H + 0.031X + 7.47, where AC is abdominal circumference, H is height, BL is body length, R is resistance, RC is rib cage circumference, W is body weight; and X is reactance. Results for TBW1 (R21 = 0.843) and TBW2 (R22 = 0.816) were highly correlated with the TBWd. When the equations were validated with data from the remaining 10 dogs, the respective mean differences between TBWd and TBW1 and TBW2 were 0.17 and 0.11 L, which equated to a nonsignificant underestimation of TBW content by 2.4% and 1.6%, respectively. Applying the two equations to dogs of various breeds showed they are inaccurate to estimate TBW content.Conclusions and Clinical Relevance—Results indicated that impedance and morphological data can be used to accurately estimate TBW content in adult Beagles. This method of estimating TBW content is less expensive and easier to perform than is measurement of TBWd, making it appealing for daily use in veterinary practice. However, the proposed equations need to be modified including morphological parameters such as body size and shape in a first approach. As in humans, morphological-specific equations have to be developed and validated.

La nutrition est un élément clé du maintien de la performance opérationnelle chez le chien de service, lequel fait souvent face à des périodes de travail intense suivies de périodes de récupération de courte durée, l’enchaînement des efforts se répétant fréquemment sur plusieurs jours. Afin d’optimiser la performance, d’éviter l’émergence de pathologies liées à l’effort physique et aux contraintes mentales, et d’améliorer la récupération, l’alimentation est maintenant intégrée de manière prépondérante dans la préparation des athlètes de haut niveau que sont ces chiens. Les objectifs de ce travail furent de : (1) concevoir un supplément nutritionnel de mise/maintien en condition et en étudier les impacts biologiques lorsqu’il est distribué avant et pendant l’effort physique, en s’intéressant plus particulièrement à son adaptation au métabolisme spécifique de l’effort demandé ; (2) concevoir un supplément nutritionnel de récupération et en étudier les impacts biologiques lorsque distribué après l’effort physique en s’axant sur les capacités de récupération à court terme. Le premier supplément développé dans cette thèse se présente sous forme extrudée spécialement conçu, et contenant des acides gras à chaînes courtes et moyennes, associés à un complexe antioxydant. Distribué une heure avant, et au bout de vingt minutes d’effort dans un protocole de course en continu chez le chien de recherche de personnes, il a généré une amélioration de la réponse de l’organisme à l’exercice via un impact positif sur les paramètres physiologiques cardiovasculaires, l’augmentation des triglycérides circulants en début d’effort, et la moindre expression plasmatique de marqueurs pro-inflammatoires et du stress oxydatif dans le groupe supplémenté. Notre deuxième étude, portant sur une supplémentation post-effort chez le chien militaire, a pour objectif d’observer l’impact d’un mélange de maltodextrines (1,5g/kg de poids corporel) et protéines auparavant sélectionnée (0,3g/kg de poids corporel) distribué immédiatement après effort, durant trois périodes d’exercice intense séparées par une heure de repos au cours d’une même journée. Nos résultats montrent une moindre augmentation plasmatique des marqueurs de l’inflammation et des témoins de lésions musculaires dans le groupe supplémenté. Des études complémentaires sont indispensables à envisager pour adapter la supplémentation à chaque typologie d’effort (endurance, mixte, résistance), en testant différents apports qualitatifs et quantitatifs. Elles permettront également de préciser les moments optimaux de distribution de celle-ci (prise unique ou fractionné, délai par rapport à l’effort), et de l’adapter aux conditions environnementales (en particulier les climats chauds, sources de nombreux problèmes chez les chiens de service projetés en opérations extérieures). / Nutrition is a key point in order to maintain performance during operationnal missions in the service dog, who often faces intense periods of work, followed by short resting periods, with a high number of working periods during several consecutive days. To optimise performance, reduce pathologies related to an intense physical exercise and mental stress and improve recovery, nutrition is now considered as one of the most important point in those elite athletes. This work objective were : (1) to develop a nutritionnal supplementation to set and maintain physical conditions when distributed before and during physical exercise, focusing our approach on the necessary adaptation to the specific metabolism involved during the exercise ; (2) to develop a nutritionnal supplementation for the recovery period, distributed just after the exercise and study the consequences of its distribution on biological markers during short term recovery period. The first supplement develop in this work contain short and medium chains fatty acids associated with an antioxidant complex. Given one hour before the work, and after a twenty minutes run during a continuous running test performed on search and rescue dogs, this supplement shows positive impacts on cardiovascular parameters, an increase of triglycerids at the onset of exercise, a reduction of plasmatic expression of proinflammatory and oxidative stress markers in the supplemented group. Our second study focusses on postexercise nutritionnal supplementation in the military dog, in order to observe impacts on a mix of maltodextrins (1.5g/kg body weight) and proteins (0.3g/kg body weight) provided just after the exercise, during three periodsof intense exercise separated by one hour rest periods during a single day. Our results include a reduction of plasmatic inflammatory markers production and of mucular lesions markers in the supplemented group. Further studies are required in order to adapt nutitionnal supplementation to each type of exercise (endurance, mix, resistance), by testing different qualitative and quantitative inputs. They will also permit to precise optimal timing for its distribution (single or fractionned inputs, timing before or after the exercise), and to adapt it to environmental conditions (especially hot climate, that is a cause of numerous problems for service dogs sent to oversees operations).