Throughout gestation, the blood vessel network of the placenta is formed sequentially by processes known as vasculogenesis and angiogenesis, which together meet the needs of the growing fetus. Normal placental angiogenesis is critical to support adequate fetal growth and assure the health of the offspring. Proper angiogenesis requires precise regulation of expression of agents that modulate this process; otherwise, pathologies of pregnancy such as preeclampsia may occur. The placenta is composed of different layers of tissue, including the lamellar (LZ), junctional, and glandular zones, each with a vascular morphology attuned to its function. We hypothesized that higher expression of pro-angiogenic factors is associated with increased morphological metrics in the LZ, the major vascularized zone. Thus, we aimed to characterize the major changes in morphology and vascular development in the placenta throughout pregnancy in cats, alongside a compressive analysis of the expression of major angiogenic factors and their receptors in the placenta, with an emphasis on the identification and interaction of different isoforms of the VEGF family. Microscopic analysis of tissue specimens from different stages of pregnancy revealed increased thickness of the LZ, especially during early to mid-gestation, at which time the tissue is composed of abundant materno-fetal interdigitations that appears rich in capillaries. VEGF proteins were detected in placental tissue in both fetal and maternal cells of the placenta, suggesting stimulatory interactions between different cell types to promote growth and angiogenesis. Gene expression analysis of placenta revealed upregulation of the pro-angiogenic factor VEGF-A in mid-pregnancy, followed by a steady decline toward term, consistent with morphologic changes in the LZ. In contrast, another pro-angiogenic factor, PlGF, showed a marked increase toward term; Flt-1, which acts as a receptor or reservoir for PLGF and VEGF A, was also upregulated at late pregnancy. Increased ratios of PLGF:VEGF-A may contribute to LZ proliferation in the last trimester. These findings are consistent with the creation of a proangiogenic placental state during gestation. Overall, we expect that this research will help elucidate mechanisms of placental vascularization, which can be applied to the design of improved strategies to treat vascular complications of pregnancy. Lastly, we applied the tools developed for placental studies to investigate pathologic angiogenesis in cutaneous squamous cell carcinoma (CSCC), a common skin cancer with major economic and medical impacts in humans and veterinary species. The creation of a new blood supply is essential for growth and metastasis of many tumor types. The goal of this study was to measure expression of variants of proteins that stimulate angiogenesis or transmit an angiogenic stimulus in feline CSCC. The results were mixed, with differences detected in expression of some regulatory agents and, for others, unexpectedly lower expression in CSSC compared to controls. Interestingly, the expression of VEGF-A relative to the protein that transmits its signal (KDR) was elevated in CSCC, suggestive of an altered signaling relationship. This finding supports our hypothesis and is consistent with human SCC studies. Our results encourage further studies on angiogenic factor variants in feline CSCC. / PHD

Hybrid male sterility (HMS) is typically the first mechanism fortifying reproductive isolation resulting from genomic incompatibilities. Three interspecies feline breeds derived from domestic cat crosses to wild cat species (Asian leopard cat and African serval) manifest HMS through several generations of backcrossing before eventually regaining fertility. This work utilized 199 hybrid individuals with varying fertilities in a genome wide association study (GWAS) comprising 63,000 genome wide SNPs. Leveraging these results with whole-testis transcriptome sequencing and quantitative real-time PCR data facilitated the comparison of transcripts in sterile and fertile hybrids. This dissertation describes four loci with highly significant and fifty with moderately significant association to sterility within each individual hybrid domestic breed and combinations of breeds. These associations help identify epistatic targets for hybrid incompatibility contributing to sterility. Comparative QTL mapping between pairs of species provides a framework to describe the accumulation of clade-specific reproductive isolating loci. Detailed exploration of gene misregulation between domestic and hybrid individuals, as well as between littermate hybrids of varying fertilities outlines a pattern of expression consistent with a meiotic sex-chromosome inactivation failure in early generations and apoptotic failure in later hybrid generations. Combining comparative genomic association and transcriptomic characterization among hybrid felids of varying divergence, new insight is gained into the mechanisms of mammalian reproductive isolation.

La prédation exercée par le Chat domestique (Felis silvestris catus) joue un rôle majeur dans la conservation de certaines espèces proies et la prévention de zoonoses. L'objectif de cette étude était d'identifier les facteurs de variation de cette prédation en milieu rural. L'étude a été conduite dans les Ardennes. La population étudiée comptait 143 individus, dont 44 % de chats de propriétaire (nourris ad libitum, 80 % stérilisés) et 56 % de chats de ferme (peu nourris, non stérilisés). Le DNA metabarcoding et le génotypage ont permis de détecter la fréquence d'apparition des aliments dans des fèces collectées sur le terrain et d'attribuer ces fèces à l'une ou l'autre des catégories de chats. Des individus ont été équipés de colliers GPS et d'accéléromètres tridimensionnels. L'éthogramme accélérométrique d'une séquence de chasse a été construit afin de pouvoir, à terme, relever les événements de prédation et de consommation des proies des individus équipés. Enfin, des variables associées à la prédation ont été mise en relation avec leur sociabilité envers les humains. Les chats ont essentiellement consommé des rongeurs et des aliments d'origine anthropique. En été, les chats de propriétaire chassaient aussi fréquemment que les chats de ferme, tandis qu'en hiver ils chassaient beaucoup moins souvent que ces derniers. Les chats de propriétaire ont de plus petits domaines vitaux que les chats de ferme. Les deux catégories de chats ont un rayon d'action restreint lorsque la pluviométrie est élevée et ont pour proies principales Microtus arvalis et Arvicola terrestris. Le degré de sociabilité des chats vis-à-vis des humains semble affecter leur succès de capture diurne de proies. / The predation exerted by the Domestic cat (Felis silvestris catus) is of special concerns for conservation and prevention of zoonosis issues. This study aims to identify the factors of variation of the predation exerted by cats in a rural landscape. It has been conducted in the Ardennes region. The population studied was composed of 143 individuals, including 44% of house cats (fed ad libitum, 80% sterilized) and 56% of farm cats (poorly fed, not sterilized). DNA metabarcoding and genotyping technics allowed to detect occurrences of food items in field-collected feces and to attribute those feces to one or the other cat categories. Some individuals were equipped with a collar composed by a GPS unit and a tri-axial accelerometer device. The ethogram of a hunting sequence based on accelerometry was constructed to gather information concerning the occurrences of predation and the consumption of the preys by the equipped individuals. Finally, some variables associated to the predation were put in relation with the sociability of cats towards humans. The cats essentially consumed rodents and anthropogenic food. During the summer, the house cats were hunting as frequently as the farm cats, whereas during the winter they were hunting a lot less comparing to the farm cats. Both cat categories have smaller daily movement range during high rainfall periods and their main preys are Microtus arvalis and Arvicola terrestris. The sociability degree of the cats towards humans seems to affect their diurnal stalk success.

Data derived largely from ecclesiastical (mostly churchwardens') accounts, foxhunting statistics, local scientific society records and 19th and 20th century literature sources from a wide range of published material, have provided detailed evidence of the status and changes in distribution over the past four centuries in Yorkshire for fox (Vulpes vulpes), badger (Meles meles), otter (Lutra lutra), pine marten (Martes martes), stoat (Mustela erminea), weasel (M. nivalis), polecat (M. putorius), American mink (M. vison), wildcat (Felis silvestris) and domestic cat (Felis catus). In the case of the domestic cat, questionnaire surveys quantified population sizes and predatory activity in rural, suburban and urban situations. Evidence of the former distribution of all the carnivores studied provides a credible historical basis for biodiversity action planning and the substantial archived database and bibliography provide further research opportunities.

La toxoplasmose, dûe au parasite Toxoplasma gondii, est une zoonose dite à cycle complexe car le pathogène fait intervenir plusieurs espèces pour assurer sa transmission. Les félidés sont les hôtes définitifs de T. gondii et, lorsqu’ils sont infectés, peuvent excréter des millions d’oocystes dans l’environnement. L’ensemble des animaux à sang chaud, y compris l’homme, constituent les hôtes intermédiaires. L'infection des différents hôtes s'effectuent par transmission verticale ou via l'ingestion de tissus animaux contaminés ou d'oocystes présents dans l'environnement.Les objectifs de ma thèse étaient i) de mesurer la fréquence et la distribution spatiale des oocystes de T. gondii dans l’environnement en milieu rural, ii) d'estimer la prévalence et la distribution spatiale de l’infection dans la communauté locale de rongeurs, iii) de déterminer les principaux facteurs responsables de la structuration spatiale de la contamination environnementale et enfin, iv) d’évaluer l'importance de l'environnement en tant que source de contamination tant pour les animaux que pour l’homme.Nous avons mis en évidence une forte contamination des sols (29%) distribuée sur l'ensemble de la zone et, avons révélé un gradient spatial similaire de diminution de la contamination avec la distance aux bâtiments au niveau des sols et dans la communauté de rongeurs. Le modèle de simulation multi-agents a permis d'expliciter le rôle prépondérant de la configuration de l'habitat humain, qui de par son effet sur la structuration spatiale des populations de chats domestiques, détermine la fréquence et la distribution des points chauds de contamination. Par conséquent, en milieu rural, l'importante contamination des sols au niveau des fermes suggère que le risque d’infection pour l’homme est élevé, indirectement à travers la consommation de viande issue d’animaux d’élevage infectés, mais aussi directement via l’ingestion d’oocystes présents sur les substrats manipulés lors de diverses activités. / Toxoplasmosis, caused by the parasite T. gondii, is a zoonosis with a complex life cycle as the pathogen requires several different species to achieve it cycle. Felids, in particular domestic cats, are the definitive hosts of the parasite and when infected they can shed millions of oocysts in the environment. All warm blooded animals, including humans, are potential intermediate hosts. Host species can be infected through vertical transmission or by ingesting contaminated tissues or oocysts present on environmental substrates.My goals were: I) to precisely measure the frequency and spatial distribution of the environmental contamination to T. gondii in a rural area, ii) to estimate the prevalence and the spatial distribution of the parasite in the local community of rodents, iii) to identify the main factors driving the spatial structure of the environmental contamination and finally, iv) to assess the importance of the environment as a transmission source for animals and humans.First, we found a high frequency of contaminated soil samples (29%) that were largely distributed across the whole area, and, we found a similar spatial gradient of decreasing contamination with increasing distances from buildings for soils and rodents. Altogether, the results obtained allowed to identify and rank the determinants of the spatio-temporal dynamics of the environmental contamination to T. gondii. The agent-based model showed the primary role of the spatial configuration of human habitat, which, through its impact on the spatial structure of domestic cat populations, determines the frequency and distribution of the hot spots of soil contamination. Thus, in rural areas, the high level of contamination within and around agricultural buildings suggests that infection risks for humans are important, either indirectly through the consumption of contaminated meat or directly due to the ingestion of oocysts contaminating earth, water or vegetables.

In der Forschung dient die Hauskatze als Modellspezies für die Entwicklung und Verbesserung assistierter Reproduktionstechniken (ART) für wildlebende Katzenarten, von denen die meisten der 36 Arten auf der Roten Liste für bedrohte Tierarten als gefährdet eingestuft sind. Eine erfolgreich angewandte Methode der ART bei Hauskatzen ist die In-vitro-Produktion von Embryonen. Allerdings gestaltet sich die Übertragbarkeit der Protokolle auf andere Feliden aufgrund speziesspezifischer reproduktionsbiologischer Besonderheiten oft sehr schwierig. Das Ziel dieser Doktorarbeit war daher die Charakterisierung des felinen Oviductins, ein Glykoprotein, welches ausschließlich im Eileiter exprimiert wird und das in mehreren Studien bei anderen Tierarten einen positiven Effekt auf die Interaktion der Gameten, Befruchtung, Teilungsraten und Embryonalentwicklung gezeigt hat. Die feline Oviductinsequenz konnte in der vorliegenden Arbeit vollständig identifiziert werden. Sowohl auf mRNA- als auch auf Proteinebene zeigte sich eine zyklusabhängige Expression von Oviductin im Eileiter mit höchsten mRNA-Kopienzahlen bzw. höchster Proteinsynthese während des Östrus. Für funktionelle Studien wurde eine Methode zur Herstellung eines rekombinant exprimierten Oviductins in E.coli etabliert. Die Anwesenheit des rekombinanten Proteins führte zu einer signifikant höheren Spermienbindung an die Zona pellucida feliner Eizellen. Die Zugabe des rekombinanten Proteins während der In-vitro Fertilisation (IVF) hatte keinen Einfluss auf die Teilungs- oder Entwicklungsraten der Embryonen, allerdings zeigten Blastocysten, die sich aus einer IVF mit Oviductin entwickelten, eine signifikant höhere Expression des gap junction protein alpha 1 Gens, was für eine verbesserte Embryonenqualität spricht. / The domestic cat serves as a model to develop and improve assisted reproductive techniques for wild felid species from which most of them are listed on the IUCN Red List of threatened species. In-vitro embryo production is successfully applied for the domestic cat but adaption to other felids, caused by their diversity in reproductive physiology, is often difficult. The biggest challenge is to create an in-vitro environment as near as possible to in-vivo conditions within the oviduct where final maturation of gametes, fertilization and early embryo development occur. Therefore, the aim of this doctoral thesis was to characterize feline oviductin, a glycoprotein which is exclusively expressed in the oviduct. Several functional studies with other species could show an influence of oviductin on gamete interaction, fertilization, cleavage rates and embryo development. In the present work gene sequence of the feline oviductin could be identified. Highest mRNA copy numbers and protein synthesis were detected during oestrus, indicating a cycle-dependent expression of oviductin in the cat. For functional studies, a method for production of recombinant oviductin in E.coli was established. Sperm-zona binding was significantly higher when recombinant oviductin was present. Addition of oviductin during in-vitro fertilization (IVF) had no effect on cleavage, morula rates or blastocyst development. In contrast, in blastocysts developed from IVF with oviductin, we found a significantly higher mRNA expression of gap junction alpha protein 1, suggesting a higher quality of in-vitro produced embryos.

Made available in DSpace on 2016-08-15T20:31:29Z (GMT). No. of bitstreams: 1 FernandaAS_DISSERT.pdf: 1414995 bytes, checksum: e7e006bb87888d5b4777f2f259d7afd8 (MD5) Previous issue date: 2015-09-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Ovarian xenografting is an auxiliary reproductive technique that allows the conservation of germplasm of high value livestock or endangered species. The use of exogenous gonadotropins assists in developing these xenografted tissues and obtaining viable follicles for in vitro embryo production (IVEP), however this use has not been reported in xenograftings of cats ovaries with C57BL/6 SCID female mice as recipients. Thus, the aim of this study was to evaluate the response of xenografting of domestic cat ovaries to equine chorionic gonadotropin (eCG) when grafted into C57BL/6 SCID female mice. Therefore, domestic cats ovarian cortex fragments were grafted under the kidney capsule of fifteen C57BL/6 SCID mice after bilateral ovariectomy. At the end of 45 days, the female mice were divided into two groups and those who did not receive hormone induction (eCG ) were euthanized at the time of induction. Females who received hormonal induction (eCG +) were euthanized after 48 hours. All collected tissues were taken for histologic processing. The proportions between the different ovarian follicles were compared by the chi-square test. The morphometric analysis of the follicles were compared between the experimental groups by the Tukey test (primordial follicles, primary and secondary) and Kruskal-Wallis (antral follicles). Macroscopically, it was possible to observe a low number (16%) of antral follicles with more than 1mm in transplants treated with eCG. In the microscopic analysis, follicles from all categories were observed in transplants and all had normal morphology and morphometry for the studied species (Felis catus), being however observed larger primordial and primary follicles in those eCG + transplants. There was a decrease in primordial follicles percentages and an increase in subsequent categories, mainly in antral follicles of eCG + group, and this condition is proposed here characterized as Follicular Right Shift (FRS). Luteinized follicles were also observed in transplants treated with eCG. Thus, it is concluded that the treatment with eCG is effective when it comes to follicular development, but it did not show a good superovulatory response / Xenotransplante ovariano é uma técnica reprodutiva auxiliar que permite a conservação do germoplasma de espécies de alto valor zootécnico ou em perigo de extinção. O uso de gonadotrofinas exógenas auxilia no desenvolvimento desses tecidos xenotransplantados e na obtenção de folículos viáveis para produção in vitro de embriões (PIVE), entretanto esse uso não foi relatado em xenotransplante de ovários de gatas com fêmeas C57BL/6 SCID como receptora. Dessa forma, o objetivo desse trabalho foi avaliar a resposta do xenotransplante ovariano de gata doméstica à gonadotrofina coriônica equina (eCG) quando transplantados em fêmeas C57BL/6 SCID. Para tanto, fragmentos de córtex ovariano de gatas domésticas foram transplantados sob a cápsula renal de quinze camundongas C57BL/6 SCID após ovariectomia bilateral. Ao final de 45 dias, as fêmeas foram divididas em dois grupos e aquelas que não receberam indução hormonal (eCG ) foram eutanasiadas no momento da indução. As fêmeas que receberam indução hormonal (eCG +) foram eutanasiadas 48h após. Todos os tecidos colhidos foram levados para processamento histológico. As proporções entre os diferentes folículos ovarianos foram comparadas pelo teste de qui-quadrado. A análise morfométrica dos folículos foi comparada entre os grupos experimentais pelo teste de Tukey (folículos primordial, primário e secundário) e Kruskal-Wallis (folículo antral). Macroscopicamente foi possível observar um baixo número (16%) de folículos antrais com mais de 1mm nos transplantes tratados com eCG. Na análise microscópica, folículos de todas as categorias foram observados nos transplantes e todos apresentaram morfologia e morfometria normais para a espécie estudada (Felis catus), sendo, porém observado folículos primordiais e primários maiores naqueles transplantes eCG +. Houve uma redução nas porcentagens de folículos primordiais e aumento nas categorias subsequentes, principalmente nas de folículos antrais do grupo eCG +, sendo essa condição caracterizada como Follicular Right Shift (FRS). Folículos luteinizados também foram observados nos transplantes tratados com eCG. Dessa maneira, conclui-se que o tratamento com eCG foi eficaz em se tratando de desenvolvimento folicular, mas não apresentou boa resposta superovulatória

Toxoplasma gondii est le parasite responsable de la toxoplasmose, une zoonose infectant l'Homme et les autres espèces endothermes. Les fermes d'élevage sont connues pour présenter de forts risques de transmission pour les hôtes et constituent des réservoirs de la contamination environnementale en milieu rural. Cette étude visait à identifier les facteurs modulant la circulation du parasite dans les fermes dans deux compartiments du cycle: les hôtes définitifs et le sol. Pour cela, la dynamique d'infection par T. gondii dans cinq populations de chats a été suivie et modélisée par des modèles de capture-marquage-recapture « multi-événements ». Ceux-ci ont montré que les taux d'expositions des chats à T. gondii varient entre fermes et que les taux d'infection sont plus élevés en automne-hiver que durant le reste de l'année. La distribution spatiale de la contamination environnementale par T. gondii a ensuite été évaluée dans six fermes par recherche d'ADN dans des prélèvements de sols, puis mise en relation avec le patron de dépôt des fèces par les chats. Malgré une large distribution de T. gondii dans les fermes, les sites de défécation des chats constituent des zones à fort risque de transmission. Le niveau de contamination dans ces sites est dépendant de leur localisation et des caractéristiques des chats qui les utilisent, tout deux influencés par la localisation du site d'alimentation des chats dans les fermes. Cette étude montre que les fermes peuvent présenter des disparités en termes d'exposition au parasite. A l'intérieur des fermes, le patron spatio-temporel de dépôt des fèces par les chats modulent les risques d'infection pour les hommes et les animaux. / Toxoplasma gondii is the protozoan parasite responsible for toxoplasmosis, a zoonosis infecting humans and other endothermic animals. Livestock farms are considered high-risk sites for T. gondii transmission to hosts and they represent reservoirs of environmental contamination in rural areas. The aim of this study was to investigate the infection dynamics of T. gondii in cat populations by surveying five dairy farms using multi-event capture–mark–recapture models. The results showed that the rates of T. gondii infection in cats varied between farms. Infection rates were higher in autumn and winter compared to the rest of the year. The spatial distribution of soil contaminated by T. gondii on six farms was assessed using quantitative PCR on soil samples and analyzed in relation to the spatio-temporal pattern of faeces deposition by cats. Although there was wide spatial distribution of T. gondii in the soil of the farms, cat defecation sites were revealed to be high-risk areas for T. gondii transmission. The level of contamination in these sites depended on their location and characteristics of cats that use them, which were both influenced by the location of the main cats feeding site in the farms. This study demonstrated that T. gondii exposure varies from one farm to another. Infection risk is also heterogeneous within a farm and is driven by the spatio-temporal pattern of deposition of cat faeces.

For many years Swedish archeologists have stumbled upon domestic cat remains in the Viking age graves. Most of the graves in this paper come from southern Sweden and Mälaren Valley where many finds have been studied. The questions are how much of the cats is left in the grave material, what kind of grave goods were deposited with the dead, if cats are usually buried either with men or women and what the combinations of all the different animal species that cats were buried with can tell us. The theory in this work is concerning human - animal relations between the vikings and their cats with the weight on antropocentrism. The point of the mentioned theory in this paper is to provide answers to what cats could mean in the viking burial ritual context. 17 grave fields have been analysed for this work with the biggest part of them located in the Mälaren Valley regions (14 grave fields), and 3 in southern Sweden. The result of this study implies that cats in the analysed Mälaren Valley and southern Sweden graves were buried with wealthy people like aristocrats and merchants. They were also seen as exotic pets during their lifetime. The cats were usually buried with other animals like dogs, horses and chickens which all propably had a status of sacral animals during viking age. Cats' remains condition is also brought up as the felines were found either as partial or full/ almost complete skelettons. Analysis results also imply that cats were buried as often with men as with women and there are also rare cases of child burials with these animals.

Data derived largely from ecclesiastical (mostly churchwardens¿) accounts, foxhunting statistics, local scientific society records and 19th and 20th century literature sources from a wide range of published material, have provided detailed evidence of the status and changes in distribution over the past four centuries in Yorkshire for fox (Vulpes vulpes), badger (Meles meles), otter (Lutra lutra), pine marten (Martes martes), stoat (Mustela erminea), weasel (M. nivalis), polecat (M. putorius), American mink (M. vison), wildcat (Felis silvestris) and domestic cat (Felis catus). In the case of the domestic cat, questionnaire surveys quantified population sizes and predatory activity in rural, suburban and urban situations. Evidence of the former distribution of all the carnivores studied provides a credible historical basis for biodiversity action planning and the substantial archived database and bibliography provide further research opportunities.