Submitted by Michele Mologni (mologni@unoeste.br) on 2018-11-23T17:01:02Z No. of bitstreams: 1 Aline da Silveira Batista.pdf: 165848 bytes, checksum: c187bcd430f8b370d46748116a9de117 (MD5) / Made available in DSpace on 2018-11-23T17:01:02Z (GMT). No. of bitstreams: 1 Aline da Silveira Batista.pdf: 165848 bytes, checksum: c187bcd430f8b370d46748116a9de117 (MD5) Previous issue date: 2018-08-16 / The objective of this study was to compare the efficiency of different techniques to recover Toxocara spp. eggs from artificially and naturally contaminated soil samples. In the first stage, soil samples (n= 12 samples of 1g) were artificially contaminated with 100 eggs of T. canis. Three techniques were used: 1) centrifugal-sedimentation after soil filtration in a series of four sieves, using distilled water (300, 212, 90, and 63 μm); 2) centrifugal-flotation with zinc sulfate (d= 1.35 g/cm3) after sieving; and, 3) centrifugal-flotation with zinc sulfate without sieving. The recovery of eggs using technique 1 was significantly superior to the other techniques, with 78.25%, 16.33%, and 10.92% positivity, respectively. In the second stage, soil samples from 15 public squares/parks were evaluated over a period of 6 months, comparing the more efficient techniques in stage 1 (techniques 1 and 2). The recovery of eggs by technique 1 was more efficient. In the three out of the 6 months evaluated, there was a significant difference in the mean number of eggs recovered, using the technique of centrifugal-sedimentation with sieving. The technique is low cost, easy to implement, and does not require solutions which present environmental risks for its execution. / O objetivo do estudo foi comparar a eficiência de três técnicas de recuperação de ovos de Toxocara spp., a partir de amostras de solo artificialmente e naturalmente contaminadas. Na primeira etapa, amostras de solo (n= 12 amostras de 1g) foram contaminadas com 100 ovos de T. canis. Foram utilizadas três técnicas: 1) centrifugo-sedimentação após filtragem do solo em série de quatro tamises (300, 212, 90 e 63 μm); 2) centrifugo-flutuação com sulfato de zinco (d = 1.35 g/cm3) após tamisação; e, 3) centrifugo-flutuação com sulfato de zinco sem tamisação. A recuperação de ovos pela técnica 1 foi significativamente superior às demais, com 78,25%; 16,33% e 10,92% de positividade, respectivamente. Em uma segunda etapa, foram avaliadas amostras de solo de 15 praças e parques públicos de Presidente Prudente, São Paulo, durante um período de 6 meses, com comparação das técnicas 1 e 2. A recuperação de ovos pela técnica 1 mostrou ser mais eficiente. Em três meses avaliados, houve diferença significativa na média dos ovos recuperados. A técnica de tamisação centrifugo-sedimentação mostrou maior eficiência na recuperação de ovos de Toxocara spp. em relação as outras técnicas. A técnica é de baixo custo, fácil execução e não requer soluções com riscos ambientais para sua execução.

The H9N2 avian influenza virus (AIV) has emerged, spread and established itself in poultry globally, in just under 30 years. During this time, multiple reassortants of H9N2 with increased zoonotic potential have been isolated in poultry and humans, causing a major threat to the economy and global health. Curiously, H9N2 appears to be compatible with multiple Hemagglutinin (HA) and Neuraminidase subtypes, in nature. Here, the aim was to investigate the HA reassortment dynamics of the poultry adapted H9N2 AIV, in a laboratory setting. Firstly, HA subtypes from wild bird isolates were cloned, before being co-transfected with the backbone of a chicken H9N2 AIV. The rescued H9N2 reassortants were titred on cells before the replication kinetics of a subset of the HA reassortants was assessed. The cDNA sequence of seven HA subtypes induced extensive recombination in E. coli, but ultimately ten out of eleven available HA subtypes were successfully cloned. Further, the chicken H9N2 AIV was compatible with all ten HA subtypes, producing infectious viral particles after co-transfection. However, all HA reassortants displayed decreased replicative fitness in MDCK-2 cells, compared to the wild-type virus. Interestingly, HA subtypes with similar genotypes cluster into distinct HA clades and groups, but these HA clades did not correlate with the replicative fitness of the reassortants. This study suggests that poultry adapted H9N2 AIV is compatible with many HA subtypes, highlighting the importance of reducing its spread in poultry, to reduce reassortment opportunities.

The study of human palaeopathology has developed considerably in the last three decades resulting in a structured and standardised framework of practice, based upon skeletal lesion patterning and differential diagnosis. By comparison, disarticulated zooarchaeological assemblages have precluded the observation of lesion distributions, resulting in a dearth of information regarding differential diagnosis and a lack of standard palaeopathological recording methods. Therefore, zoopalaeopathology has been restricted to the analysis of localised pathologies and ‘interesting specimens’. Under present circumstances, researchers can draw little confidence that the routine recording of palaeopathological lesions, their description or differential diagnosis will ever form a standard part of zooarchaeological analysis. This has impeded the understanding of animal disease in past society and, in particular, has restricted the study of systemic disease. This research tackles this by combining the disciplines of human palaeopathology and zoopalaeopathology and focusing on zoonotic disease. The primary aim of this research was to investigate the skeletal manifestation of bTB in cattle, sheep/goat and pig to establish differential diagnostic criteria for its identification in zooarchaeological assemblages. Methods commonplace in human palaeopathology were adapted and applied to zoopalaeopathology, in addition to radiography and aDNA analysis. The results emphasise the difficulties but also the potential associated with the identification of systemic diseases in zooarchaeological assemblages. An approach to the classification of potentially infectious lesions is presented that enables the calculation of crude prevalence in disarticulated assemblages. In addition, the potential for a DNA analysis to shed further light on animal disease in the past is emphasised. / Arts and Humanities Research Council (AHRC) / Many of the images have been removed from the online version due to copyright restrictions. The embargo period for the thesis ended: 16th January 2018.

Le raton laveur (Procyon lotor) est porteur de nombreux parasites dont certains sont des agents de zoonose. Il nous est apparu opportun d’étudier la faune parasitaire intestinale du raton laveur québécois, particulièrement l’espèce Baylisascaris procyonis. L’étude a donc porté sur l’examen de matières fécales (n = 301) et de contenus intestinaux (n = 203) de 351 ratons laveurs provenant de deux programmes gouvernementaux sur la rage du raton laveur et d’excréments (n = 409) provenant de latrines dans des parcs nationaux du Québec. Des oeufs de B. procyonis étaient excrétés par 23,3 % des ratons laveurs, alors que 29,1 % étaient porteurs de vers. La charge parasitaire moyenne des vers B. procyonis de tout stade variait de 8,5 chez les adultes à 27,1 chez les juvéniles. Chez ces derniers, la charge parasitaire était 11,4 fois supérieure à celle des ratons adultes (p < 0,0001) dont 23,7 fois plus de vers immatures (p < 0,0001) et 9,1 fois plus de vers matures (p = 0,01). Le nombre d’oeufs était 24,7 fois plus élevé chez les ratons juvéniles (p = 0,02) et 50,0 fois moins élevé chez la femelle allaitante. La probabilité du raton d’excréter des oeufs l’automne était de 11,1 (IC95 %: 1,21-101,60) par rapport au printemps et de 21,1 (IC95 %: 2,91-153,18) par rapport à l’été (p < 0,01). La sensibilité (Se) et la spécificité (Sp) de la coproscopie comparée à la nécropsie (norme étalon) ont été calculées en se basant sur l’observation des vers matures (Se : 81,8 %; Sp : 97,7 %) et des vers de tout stade (Se : 53,9 %; Sp : 97,0 %). De plus, la recherche des parasites intestinaux nous a permis de découvrir des ookystes de Giardia (6/159), espèce parasitaire nouvellement rapportée chez le raton. En 2007, nous avons trouvé, par échantillonnage unique, des oeufs de B. procyonis dans 15,8 % des latrines (n = 165) identifiées dans des parcs nationaux du sud du Québec et en 2008, par échantillonnage multiple, dans 89,7 % des latrines (n = 26) actives situées dans des secteurs accessibles à l’homme dans trois parcs nationaux de la Montérégie. Le potentiel zoonotique de B. procyonis est un problème de santé publique qui pourrait devenir sérieux étant donné le pourcentage élevé de ratons laveurs qui excrètent le parasite et qui contaminent des secteurs accessibles par les humains. / Raccoons are carriers of many parasites, some of which are zoonotic. We wanted to expand our knowledge of the Quebec raccoon intestinal parasitic fauna, specifically the Baylisascaris procyonis species. The study consisted of fecal matter (n = 301) and intestinal content examinations (n = 203) collected from 351 raccoons originating from two government programs on raccoon’s rabies and excrements collected (n = 409) from latrines in Quebec national parks. Eggs from B. procyonis were excreted by 23.3 % of raccoons while 29.1 % were worm carriers. The average parasitic burden of all stages of B. procyonis worms varied from 8.5 in adults to 27.1 in juveniles. In juveniles, the parasitic burden was 11.4 times superior to adult raccoons (p < 0.0001) with 23.7 times more immature worms (p < 0.0001) and 9.1 times more mature worms (p = 0.01). The number of eggs was 24.7 times superior in juvenile raccoons (p = 0.02) and 50.0 times lower in breast-feeding females. The probability of raccoons excreting eggs in autumn was 11.1 (IC95 %: 1.21-101.60) in comparison to spring and 21.1 (IC95 %: 2.91-153.18) in comparison to summer (p < 0.01). The sensitivity (Se) and specificity (Sp) of coproscopy compared to necropsy (gold standard) were calculated based on observations of mature worms (Se: 81.8 %; Sp: 97.7 %) and worms of all stages (Se: 53.9 %; Sp: 97.0 %). In addition, our intestinal parasitic research permitted the finding of Giardia oocysts (6/159), which is a newly found parasite species in raccoons. In 2007, B. procyonis eggs were found in 15.8 % of latrines (n = 165) identified in Southern Quebec national parks and in 89.7% of active latrines (n = 26) in three Eastern Townships national parks sites accessible to man in 2008. The zoonotic potential of B. procyonis is a public health issue that could become serious considering the high percentage of raccoons excreting the parasite thus contaminating sectors accessible to man.

Cryptosporidium spp. est un protozoaire parasite du système gastro-intestinal largement répandu chez les vertébrés et causant la cryptosporidiose, une zoonose occasionnant des troubles digestifs sévères pouvant entrainer la mort chez les individus immunodéficients. Au Canada, la déclaration de cette maladie est obligatoire depuis l’an 2000. Ainsi, il est pertinent de mieux comprendre l’infection chez les animaux de compagnie, puisqu’ils sont potentiellement un réservoir du parasite. Durant l’année 2008, des échantillons fécaux provenant de 1 202 chats (n = 371) et chiens (n = 831) de la province du Québec ont été analysés par comptage des ookystes de Cryptosporidium spp. au moyen de la technique de centrifugation en solution de sulfate de zinc. Dans cette étude,la prévalence de Cryptosporidium spp. chez les chats (28/371 : 7,55 %) et chez les chiens(88/831 : 10,59 %) de compagnie confirme leur potentiel en tant que réservoir du parasite. Au Québec, de par leur nombre, les chats sont potentiellement un réservoir zoonotique du parasite plus important que celui des chiens, bien qu’il n’existe pas de différence significative entre la prévalence du parasite chez le chat et le chien pour l’année 2008. L’âge (p = 0,0001) et l’infection concomitante par Giardia spp. (p = 0,0001) se sont avérés être des facteurs associés avec la présence de Cryptosporidium spp. chez le chien. Parmi l’ensemble des variables testées chez le chat (l’âge, le sexe, la saison et l’infection concomitante par Giardia spp.), aucune n’a été associée de manière significative à la présence du parasite chez le chat. Ceci peut être dû au nombre limité d’individus testés pour cette espèce. Un suivi de l’excrétion des ookystes de Cryptosporidium spp. chez deux chats suggère que l’excrétion des ookystes peut se faire sur une période de sept mois et que le taux d’excrétion varie dans le temps. Le diagnostic moléculaire des espèces et génotypes de Cryptosporidium spp. isolés à partir des échantillons de matières fécales devait être réalisé par la technique de PCR emboîtée des fragments des gènes ARNr 18S et HSP70 et du séquençage des produits de PCR. Aucun résultat positif n’a toutefois été obtenu. Afin d’augmenter la puissance statistique des analyses épidémiologiques sur la prévalence de Cryptosporidium spp., il serait nécessaire à l’avenir de travailler sur un nombre d’animaux beaucoup plus important. / Cryptosporidium spp. is a protozoan parasite from the gastro-intestinal tract with a large range of vertebrate hosts causing cryptosporidiosis, a zoonotic disease which may lead to severe digestive troubles and sometimes death for immunocompromised people. It is a noticeable disease in Canada since 2000. It is thus relevant to study the infection in cats and dogs as they may represent an important zoonotic reservoir for the parasite. A total of 1,202 stool samples from cats (n = 371) and dogs (n = 831) from the province of Quebec were examined during the year 2008 for this research. The prevalence of Cryptosporidium spp. was calculated using coprology tests using centrifugation in a zinc sulfate solution. The prevalence in cats (28/371: 7,55 %) and dogs (88/831: 10,59%) corroborates that pets living in the province of Quebec may be a reservoir for Cryptosporidium spp. While we did not find a significant difference in the prevalence of Cryptosporidium spp. between cats and dogs, cats may represent a larger reservoir as they represent a larger population within the province. We identified age (p = 0,0001) and concomitant infection with Giardia spp. (p =0,0001) as risk factors for dogs. Among all the variables tested on cats (age, sex, season, concomitant infection with Giardia spp.), none were significantly associated with the presence of the parasite in cats. This may be caused in part by the small number that was analyzed. A follow-up study with two cats showed the excretion of oocysts can last for a minimum of seven months and varies in intensity through time. The molecular diagnostic of species and genotypes of Cryptosporidium spp. isolated from fecal samples should have been done by using the amplification of the gene fragments ARNr 18S and HSP70 nested PCR reactions and the sequencing of PCR products. Although this technique was attempted in this study, no positive result was obtained. It is recommended to work on larger animal populations to increase the statistical power of epidemiological analysis of the prevalence of Cryptosporidium spp.

Orientador: Antonio Carlos Paes / Resumo: Um protocolo de coleta, extração e amplificação de ácidos nucléicos (ANs) utilizando a técnica de PCR em tempo real para detecção de Mycobacterium bovis (M. bovis) e/ou Mycobacterium tuberculosis (M. tuberculosis) foi testado em 60 linfonodos provenientes de bovinos. Desses 60, 30 foram provenientes de bovinos suspeitos de tuberculose bovina (TBb), outros 30 apresentavam lesões nodulares classificadas como adenite pelo Serviço de Inspeção Federal (SIF), na linha de abate. Os resultados obtidos demonstraram que dos 30 linfonodos coletados suspeitos de TBb, 24 (80%) foram confirmados positivos para M. bovis e/ou M. tuberculosis e seis (20%) foram confirmados negativos. Dos 30 linfonodos com a presença de lesões nodulares do tipo adenite, 13 (43,33%) foram positivos para TBb e 17 (56,66%) foram negativos para TBb. Não há um teste diagnóstico rápido e preciso para detecção da TBb, diante disso o presente estudo desenvolveu um método de diagnóstico para a TBb baseado em PCR em tempo real com análise da fluorescência do corante SYBR Green. Estes resultados indicaram que o PCR em tempo real foi específico e sensível para a detecção e identificação dos dois alvos selecionados para detectar a TBb, com melhores resultados para o DNA genômico, quando comparado com o RNA ribossômico. O teste padronizado de qPCR para o diagnóstico da TBb permitiu uma maior acurácia e significativa redução do tempo de diagnóstico em relação aos métodos tradicionais de identificação da TBb. / Abstract: A protocol for the collection, extraction and amplification of nucleic acids (ANs) using a PCR technique for detection of Mycobacterium bovis (M. bovis) and / or Mycobacterium tuberculosis (M. tuberculosis) was tested on 60 bovine-derived lymph nodes. . Of these 60, 30 were included in cattle suspected of having bovine tuberculosis (TBb), another 30 had sclerias classified as adenites by the Federal Inspection Service (SIF), at the slaughter line. The results showed that the 30 lymph nodes with suspected TBb, 24 (80%) were confirmed positive for M. bovis and / or M. tuberculosis and six (20%) were confirmed negative. Of the 30 lymph nodes with adenitis nodular lesions, 13 (43.33%) were positive for TBb and 17 (56.66%) were negative for TBb. TBB, together with the present study for a diagnostic method for TBB, is a real-time test with SYBR Green dye fluorescence analysis. Results compared with real-time PCR were specific and sensitive for the identification and identification of the two targets selected to detect a TBb, with samples obtained for genomic DNA, when observed with ribosomal RNA. The standardized qPCR test for the diagnosis of tuberculosis allowed for greater accuracy and a significant reduction in diagnosis time in relation to TBb identification parameters. / Doutor

Apesar do estado de São Paulo ter uma área de vegetação nativa pequena e, em sua maioria, altamente fragmentada, isolada e imersa em matrizes agrícolas em propriedades privadas, seu território ainda alberga um número considerável de espécies silvestres, inclusive nessas áreas comerciais. Para a elaboração de planos de manejo ecologicamente corretos, é necessário selecionar (1) espécies-chave, como os mamíferos carnívoros; (2) fatores ecológicos primários das espécies, como a dieta e o uso do espaço; e (3) locais com histórico de uso do solo representativos da exploração espaço-temporal, como, por exemplo, Angatuba (SP) em São Paulo. Essa dissertação foi desenvolvida na Fazenda Três Lagoas e seus arredores, entre maio de 2008 e junho de 2010 e abrange três temas distintos, mas interligados. No primeiro capítulo fez-se uma introdução geral sobre os temas. No segundo avaliou-se, em quatro fazendas adjacentes, a exposição de carnívoros domésticos aos agentes infecciosos da parvovirose, raiva, cinomose e leptospirose através de exames sorológicos. Verificou-se que as prevalências de contato dos 13 cães amostrados com esses agentes patológicos, foram, respectivamente, 100%, 80%, 23% e 15,4%. As reações positivas a alguns desses agentes e sorovares, mostram que os cães domésticos tiveram contato com os agentes através de outros hospedeiros, pois esses agentes não estão presentes em vacinas. Isso demostra a importância de sua detecção na vida silvestre e no ambiente visando elaborar um manejo sanitário efetivo para a economia e saúde humana e para a vida silvestre. No terceiro capítulo estudou-se a dieta de 12 espécies de mamíferos carnívoros através da análise de 524 amostras de fezes coletadas nas Fazendas Três Lagoas e Arca. Pode-se verificar que os carnívoros presentes na área possuem hábitos onívoros, com algumas alterações da dieta em comparação com áreas mais preservadas, consumindo em grande quantidade pequenos roedores, frutos e insetos. A competição, apesar de pequena, foi constadada entre as espécies domésticas e silvestres. Verificou-se, de forma complementar, que as espécies usam mais as áreas próximas da vegetação nativa que dentro da matriz, sendo que algumas espécies estiveram restritas ao ambiente nativo. Esse estudo sugere que essas paisagens albergam somente espécies com hábitos mais plásticos e que a matriz silvicultural é permeável a alguns predadores. No quarto capítulo, estudou-se a prevalência de ectoparasitas naquelas amostras de fezes, que mostrou-se de 42,17% para ácaros, dentre eles carrapatos, e 4% de pulgas. As prevalências desses artrópodes variaram entre as espécies de carnívoros. Os ácaros foram encontrados nas fezes de todas as espécies amostradas, e as pulgas estiveram presentes em fezes de todos os canídeos, de todos os felídeos, com exceção do gato-mourisco, e do quati. Foram detectados carrapatos em fezes de espécies de hospedeiros ainda não relatadas na literatura, mostrando serem hospedeiros não-usuais, o que poderia demonstrar um aumento de sua prevalência no ambiente como consequência provável da mudança da paisagem. Essa maior carga de parasitas, juntamente com o contato frequente da fauna silvestre com os humanos e seus animais domésticos, atesta a necessidade de estudos epidemiológicos de maior amplitude nessas paisagens agrícolas. / Although the state of Sao Paulo has a small area of native vegetation, mainly fragmented, isolated and highly immersed in agricultural matrices on private properties, it still lodges a considerable number of wild species, also on these commercial areas. For elaboration of ecologically correct manegement plans, it´s necessary to select (1) key-species, as carnivore mammals; (2) species primary ecological factors, as diet and space use; and (3) representative areas of space-temporal exploration of the land, as Angatuba does on Sao Paulo. This study was carried out at Fazenda Três Lagoas and its surrounding between May 2008 and June 2010, and encloses three distinct but linked subjects. The first chapter is a general introduction of those subjects. The second one evaluates, in those four adjacent farms, domestic carnivores exposition to infectious agents of parvovirosis, rabies, canine distemper and leptospirosis through serologic examinations. The prevalence of contact of those pathological agents, in the 13 dogs, were 100%, 80%, 23% and 15.4%, respectively. The positive reactions to some of these agents and serovares show that those domestic dogs had contact to other hosts, as some of them do not compose vaccines. It shows the importance of its detention in wildlife and in the environment aiming to elaborate an effective sanitary management for economy and human health and for wildlife conservation. In the third chapter the diet of 12 species of carnivorous mammals were investigated through analysis of 524 scats collected on Fazenda Três Lagoas and Fazenda Arca. Carnivore mammals on these area have omnivores\' food habits, with some modifications on diet in comparison with those in preserved areas, consuming in a great amount small rodents, fruits and insects. The competition, although weak, occurs among the domestic and wild species. In a complementary way, it was verified that the animals use more areas closer to native vegetation than inside matrix and that some species use areas restricted to native environment. This study suggests that these landscapes only lodge species with more plastic habits and that silvicultural matrix is permeable to some predators. In the forthy chapter, it was investigated the ectoparasites prevalence in those excrement samples and revealed a prevalence of 42,17% for mites, amongst them ticks, and 4% of fleas. The prevalence of those arthropods varied between carnivore species. Mites were found in excrements of all sampled species, and fleas were found in all wild dogs and wild cats faeces, but jaguarundi, and coati. Some carnivore scats contenied different ticks showing that those mammals are non-usual hosts of them and that, possibly, there is an increase of its prevalence in the environment as probable effect of the landscape modifications. A higher parasitic load with direct contact of wildlife to humans and livestock request to carry out a more in-depth epidemiologic investigation into these agricultural landscapes.

La gestion de populations de rongeurs est importante car ils peuvent causer divers problèmes économiques, écologiques et sanitaires. Cette gestion s'appuie sur des méthodes de prévention, celles-ci étant des mesures architecturales, hygiéniques et des pratiques agricoles. Elle s'appuie également sur des méthodes de lutte biologique, chimique et mécanique. La lutte chimique se fait principalement avec l'utilisation de rodenticides anticoagulants qui agissent plusieurs heures après ingestion permettant d'éviter l'éviction alimentaire. L'utilisation excessive de ces molécules a entrainé la sélection de résistance chez certains individus rendant leur utilisation peu efficace. Il y a alors eu le développement de nouvelles molécules, mais celles-ci sont plus écotoxiques. Nous nous sommes intéressés dans ces travaux de thèse à la gestion de populations de rongeurs dans un cadre de risque sanitaire leptospirosique. Les rongeurs sont les principaux réservoirs de leptospires pathogènes, des bactéries Spirochètes responsables de la leptospirose. La leptospirose est une zoonose ré-émergeante à répartition mondiale dont le nombre de cas annuel est évalué à approximativement un million et dont il y a eu une augmentation du nombre de cas ces dernières années. Elle est endémique en régions tropicales. C'est une maladie dont l'incidence réelle est sous-estimée en raison notamment de la grande variabilité de symptômes possibles allant du syndrome grippal à des formes rénales. Le taux de mortalité est élevé aux alentours de 10% mais pouvant aller jusqu'à 50% dans certains cas avec des complications pulmonaires. De nombreux facteurs influent sur le cycle complexe de transmission de la leptospirose. Il peut y avoir un risque sanitaire lié à la présence de rongeurs porteurs de leptospires pathogènes, qui peut être difficile à contrôler si les rongeurs sont résistants aux rodenticides anticoagulants utilisés pour réguler leurs populations. De plus, il peut y avoir des problèmes d'exposition à des risques écotoxiques lors de mise en place de traitements chimiques. Nous avons étudié plusieurs cas concrets montrant la difficulté dans la prise de décision liée à la gestion des rongeurs dans des contextes divers. Dans un contexte de problèmes économiques, nous avons analysé le portage de leptospires pathogènes par des campagnols des champs et des campagnols terrestres ainsi que la résistance de cible à la bromadiolone, molécule utilisée en lutte chimique à basse densité de population. Nous nous sommes intéressés à l'influence de la perception sociétale sur le risque sanitaire lié à des souris dans un élevage bovin dans lequel il y a des cas de leptospirose humaine et de leptospirose bovine. Dans le cadre d'un projet visant à étudier l'efficacité de gestion de populations de rongeurs commensaux en Martinique dans un contexte leptospirosique, nous avons regardé l'exposition des rongeurs aux AVKs et la résistance de cible aux AVKs. Pour le portage de leptospires pathogènes par les campagnols nous avons trouvé des prévalences allant de 3% à 53% selon la zone étudiée. Il y avait 3 mutations sur le gène vkorc1 amenant peu de résistance de cible à la bromadiolone. Dans l'élevage bovin nous avons notamment trouvé que la perception sociétale de l'éleveur sous-estimait le risque lié aux souris dont 41% étaient porteuses de leptospires pathogènes et 50% étaient porteuses de mutations du gène vkorc1 amenant de la résistance de cible aux AVKs. En Martinique nous avons trouvé que 54% des individus analysés avaient été exposé à au moins une molécule AVKs et qu'il y avait plusieurs mutations du gène vkorc1 dont certaines pas encore caractérisées amenant de la résistance de cible aux AVKs. La gestion des rongeurs est un problème complexe avec plusieurs facteurs à prendre en compte que ce soient les risques liés à l'exposition aux rongeurs ou ceux liés à l'utilisation de rodenticides anticoagulants [etc...] / Management of rodent populations is important because they can cause various economic, ecologic and health problems. This management relies on prevention methods, these ones being architectural, hygienic and agricultural practices. It relies on biologic, chemical and mechanic control methods. Chemical control methods are made mainly with the use of anticoagulant rodenticide acting several hours after ingestion to provide food eviction. Excessive use of these molecules led to resistance selection of some individuals yielding their use underperforming. Then there has been new molecules development, but they are more ecotoxic. In these thesisworks we are interested in management of rodent populations within a framework of health risk of leptospirosis. Rodents are main reservoirs of pathogenic leptospires, Spirochete bacterias responsible of leptospirosis. Leptospirosis is a worldwide re-emerging zoonosis of which there are approximately one million of cases each year and of which there is an increasing number of cases these last years. It is endemic in tropical regions. It is a disease of which real incidence is underestimated because of especially large variability of possible symptoms ranging from flu syndrome to renal forms. Mortality rate is high around 10% and up to 50% in certain cases with pulmonary complications. Numerous factors influence on complex transmission cycle of leptospirosis. There can be health risk associated with presence of rodent carriers of pathogenic leptospires, which can be difficult to control if rodents are resistant to anticoagulant rodenticides used to regulate their populations. Furthermore, there can have exposition problems to ecotoxic risks throughout establishment of chemical treatment. We studied several concrete cases showing the difficulty in decision-making relied with rodent management in various contexts. In a context of economic problems, we analysed pathogenic leptospires carriage by meadow voles and ground voles as well as target resistance to bromadiolone, used molecule in chemical control of low population density. We are interested in influence of societal perception on health risk relied with mice in bovine farming in which there are human and bovine leptospirosis cases. In framework of a project leading to study efficiency of commensal rodent populations management in Martinique in a leptospirosis context, we looked rodent exposition to AVKs and target resistance to AVKs. For pathogenic leptospires carriage by voles we found prevalence ranging from 3% to 53% according to studied area. There were 3 mutations on vkorc1 gene leading to few target resistances to bromadiolone. In the bovine farming we found especially that societal perception of the breeder underestimated risk relied with mice of which 41% were carriers of pathogenic leptospires and 50% were carriers of vkorc1 gene mutations leading to target resistance to AVKs. In Martinique we found that 54% of analysed individuals have been exposed to at least one AVKs molecules and that there were several vkorc1 gene mutations in which certain were not characterised yet leading to target resistance to AVKs. Rodent management is a complex problem with several factors to take into account whether risks relied with rodent exposition or those relied with the use of anticoagulant rodenticides. Furthermore, the concept of risk is scientific but the perception which is subjective participates in decision-making of a way of management

Le protozoaire Blastocystis et la bactérie Campylobacter sont des microorganismes zoonotiques ayant un impact majeur en santé publique. Même si son pouvoir pathogène reste controversé, Blastocystis est l’eucaryote unicellulaire le plus fréquemment retrouvé dans les selles humaines alors que Campylobacter est considérée comme la cause la plus courante de gastroentérite humaine. Ces microorganismes sont fréquemment présents chez les animaux destinés à l’alimentation comme les volailles ou les bovins et leur transmission peut être liée à la consommation ou à un contact répété avec les manipulateurs de ces animaux. Malgré ce risque potentiel majeur pour l’Homme, très peu de données étaient disponibles concernant la prévalence et les espèces de Campylobacter ou sous-types de Blastocystis présents dans les filières avicole et bovine au Liban. De plus, le risque de transmission zoonotique par contact direct avec le personnel des abattoirs ou les éleveurs n’avait jamais été évalué. Pour ce qui est des bovins, Blastocystis a été identifié par PCR quantitative en temps réel dans plus de 60% des échantillons de selle de vaches laitières analysés au Nord-Liban. Une prédominance du sous-type (ST) 10 et du ST14 a été observée confirmant que les bovins sont les hôtes naturels de ces deux STs. Les ST2, ST1, ST5, ST3 et ST7 ont aussi été identifiés avec de faibles prévalences. Chez les éleveurs comme dans un groupe de patients sans contact avec ces animaux, une prévalence importante dépassant les 50% a été rapportée avec une prédominance du ST3. Cependant, la comparaison des isolats identifiés dans ces cohortes humaine et animale suggère que les bovins joueraient un rôle négligeable en tant que réservoirs zoonotiques de Blastocystis. Dans la filière avicole, la prévalence de ce même parasite dépassait les 30% dans les caeca de poulets de chair analysés dans des abattoirs du Nord-Liban. Tous les isolats aviaires caractérisés appartenaient aux ST6 et ST7 confirmant que les oiseaux sont des hôtes naturels de ces deux STs. Chez le personnel d’abattoir comme dans un groupe de patients sans contact avec les volailles, la prévalence de Blastocystis dépassait les 50% avec une prédominance du ST3. De plus, l’identification du ST6 aviaire chez le personnel d’un abattoir confirmait le potentiel zoonotique de ce ST. Toujours dans cette filière avicole et en analysant à la fois les même caeca et des carcasses de poulets en fin de chaîne d’abatage, la prévalence de Campylobacter était respectivement de 67% et 17,2% dans ces prélèvements. Les deux principales espèces identifiées étaient C. jejuni et C. coli. Des niveaux élevés de diversité génétique ont été observés parmi les 51 isolats de C. jejuni identifiés chez les poulets et répartis en 25 profils distincts. Une prédominance des profils 1, 13, 30 et 38 a été observée alors qu’ils sont aussi fréquemment retrouvés parmi les cas cliniques humains français suggérant que ces animaux représentent un réservoir potentiel de campylobactériose humaine. Une prévalence significativement plus élevée de Campylobacter a été rapportée chez le personnel de l'abattoir par rapport à celle observée dans la cohorte de patients sans contact avec la volaille montrant que les poulets de chair contaminés à l'abattoir représentaient une source non négligeable de transmission zoonotique de la bactérie. Les mêmes caeca de poulets ont pu être comparés pour la présence de Campylobacter et de Blastocystis. Cette comparaison révélait une association statistiquement significative de ces deux microorganismes suggérant que la présence de Campylobacter serait favorisée par celle de Blastocystis et vice versa [...] / The protozoan Blastocystis and the bacterium Campylobacter are zoonotic microorganisms with a major impact on public health. Although its pathogenicity remains controversial, Blastocystis is the most common single-celled eukaryote found in human stool, while Campylobacter is considered the most common cause of human gastroenteritis. These microorganisms are frequently present in food-producing animals such as poultry or cattle and their transmission may be related to consumption or repeated contact with the handlers of these animals. Despite this major potential risk to humans, very little data were available on the prevalence and species of Campylobacter or Blastocystis subtypes present in the poultry and bovid sectors in Lebanon. In addition, the risk of zoonotic transmission through direct contact with slaughterhouse staff or farmers had never been assessed. Regarding bovids, Blastocystis was identified by PCR in more than 60% of the dairy cattle stool samples analysed in North Lebanon. A predominance of ST10 and ST14 has been observed confirming that cattle are the natural hosts of these two STs. ST2, ST1, ST5, ST3 and ST7 were also identified with lower prevalences. In breeders as well as in a group of patients without contact with these animals, a significant prevalence exceeding 50% has been reported with a predominance of ST3. However, a comparison of the isolates identified in these human and animal cohorts suggests that cattle would play a negligible role as zoonotic reservoirs of Blastocystis. In the poultry sector, the prevalence of the same parasite exceeded 30% in broiler caeca analysed in slaughterhouses in North Lebanon. All characterized avian isolates belonged to ST6 and ST7 confirming that birds are natural hosts of these two STs. Among slaughterhouse staff as well as in a group of patients without contact with poultry, the prevalence of Blastocystis exceeded 50% with a prevalence of ST3. In addition, the identification of avian ST6 in slaughterhouse staff confirmed the zoonotic potential of this ST. Still in this poultry sector and by analysing both the same caeca samples and chicken carcasses, the prevalence of Campylobacter was 67% and 17.2%, respectively, in these specimens. The two main species identified were C. jejuni and C. coli. High level of genetic diversity was observed among the 51 C. jejuni isolates identified in chickens and distributed in 25 distinct profiles. A predominance of profiles 1, 13, 30 and 38 has been observed, although they are also frequently found among French human clinical cases suggesting that these animals represent a potential reservoir of human campylobacteriosis. A significantly higher prevalence of Campylobacter was reported among slaughterhouse staff compared to that observed in the cohort of patients without contact with poultry, showing that contaminated broilers at the slaughterhouse were a significant source of zoonotic transmission of the bacteria. The same chicken caeca samples were compared for the presence of Campylobacter and Blastocystis. This comparison revealed a statistically significant association of these two microorganisms suggesting that the presence of Campylobacter would be enhanced by that of Blastocystis and vice versa. This first large-scale survey conducted in Lebanon provided major data on the prevalence and circulation of Blastocystis and Campylobacter in this ocuntry and assessed the need to put in place measures to prevent and control these microorganisms in poultry and cattle breeding or slaughter facilities in order to limit their transmission.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2016-08-05T13:52:22Z No. of bitstreams: 1 2010 - Renata Santiago Alberto Carlos.pdf: 1449902 bytes, checksum: e7a95db4b4c56f516c350e9c2b885966 (MD5) / Made available in DSpace on 2016-08-05T13:52:22Z (GMT). No. of bitstreams: 1 2010 - Renata Santiago Alberto Carlos.pdf: 1449902 bytes, checksum: e7a95db4b4c56f516c350e9c2b885966 (MD5) Previous issue date: 2010-02-22 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico, CNPq. / Toxoplasma gondii is an obrigatory intracelular parasite that has the felids as definitive host (DH) and the homoeothermic animals as intermediary hosts (IH). The infection occurs by ingesting sporulated oocysts that were eliminated in DH feces, by bradyzoites when ingested by IH meat or by tachyzoites when occur vertical transmission. In this study, 529 canine blood samples were collected, 120 at the municipality of Itabuna, and 409 at the municipality of Ilh?us. All blood samples of Itabuna were consisted of animals from domiciled urban areas. From Ilh?us, 96 were domiciled from urban areas, 135 were urban but strayed dogs from Centre for Zoonosis Control (CZC) and 178 were domiciled and from rural area. The Indirect hemagglutination test was used to test the presence of anti-T. gondii and a questionnaire were applied to the domiciled dog owners to analyze risk factors associated to the infection. From all 529 dogs, 193 were positive (36.5%). At Ilh?us 37.7% (154/255) and at Itabuna 32.5% (39/81) of animals were positive. The risk factors associated to the infection were localization, rural dogs had higher chance than urban (p=0.001), the behavior, the strayed urban dogs had more chance than the urban domiciled dogs (p=0.01), home-made food and meat ingestion (p= 0.034 and 0.027 respectively), race, the undefined race dogs had more risk (p=0.0001) and age, were the animals under five years old had a higher risk to the infection (p=0.009). The presence of veterinary assistance was considered as a protection factor. The logistic regression confirmed the habitat and age of the animals as the main risk factor associsted with the infection. / Toxoplasma gondii ? um parasito intracelular obrigat?rio que tem como hospedeiro definitivo membros da fam?lia Felidae e animais homeot?rmicos como hospedeiros intermedi?rios. A infec??o se d? pela ingest?o de oocistos esporulados eliminados nas fezes dos felinos, pela ingest?o de bradizo?tos nos tecidos dos HI ou ainda pela transmiss?o vertical de taquizo?tos. Neste estudo coletou-se sangue de 529 c?es, sendo 120 do munic?pio de Itabuna e 409 de Ilh?us. Todos os animais de Itabuna eram urbanos domiciliados. Dentre os c?es de Ilh?us, 96 eram urbanos domiciliados, 135 eram urbanos errantes provenientes do Centro de Controle de Zoonoses (CCZ), e 178 eram rurais domiciliados. Foi realizado o Teste de Hemaglutina??o Indireta (HAI) para analisar a presen?a de anticorpos anti-T.gondii e um question?rio foi aplicado junto aos propriet?rios dos animais domiciliados para avalia??o dos principais fatores de risco associados ? infec??o. Do total de 529 c?es, 193 foram positivos (36,5%). No munic?pio de Ilh?us 37,7% (154/255) e em Itabuna 32,5% (39/81) foram positivos. Os fatores de risco associados ? infec??o foram o habitat, onde os animais rurais tiveram maior risco do que os urbanos (p=0,001), o modo de vida, em que os animais urbanos errantes tiveram maior risco do que os urbanos domiciliados (p=0,01), ingest?o de comida caseira e de carne (p= 0,034 e 0,027, respectivamente), ra?a apresentando os SRD com maior risco (p=0,0001) e idade onde os animais acima de cinco anos tiveram maior risco (p=0,009). A presen?a de orienta??o do M?dico Veterin?rio se mostrou fator de prote??o (p=0,002). A regress?o log?stica confirmou como principais fatores de risco a idade e o habitat dos animais do estudo