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Almeida.pdf: 2582224 bytes, checksum: 619daa8290bd853bcd6681edd48defdf (MD5) Previous issue date: 2017-07-28 / FAPEAM - Fundação de Amparo à Pesquisa do Estado do Amazonas / The species of the family Lamiaceae are known in Brazil for being very aromatic and for presenting bioactive properties as antioxidant, bactericide, fungicide and insecticide. In this work was analyzed Hyptis dilatata Benth, found in areas of cerrado in Brazil. The chemical components, the biological activity of the essential oil and extracts of leaves and flowers, considering dry and rainy periods, of specimens of the Serra do Tepequém, in Roraima-Brazil, were studied. The essential oils were extracted from the leaves collected at different times (morning, afternoon and night) and from the flowers in the dry and rainy periods. In relation to the hexane and ethanolic extracts from leaves and flowers, these were extracted from specimens collected during the dry and rainy season. The results of the phytochemical profile, derived from the analysis of the hexane and ethanolic extracts of leaves and flowers, presented different secondary metabolites, being identified coumarin, flavonoids, phenols, saponins, alkaloids, among others. The chemical characterization of the essential oils of leaves and flowers showed as main substances α-pinene, β-pinene, limonene, 3-carene, β-caryophyllene, fenchone and camphor. The essential oil extracted from the flowers in the rainy and dry period inhibited the enzyme acetylcholinesterase, respectively, 93.4% and 92.4%, whereas the essential oil extracted from the leaves, in the dry period and in the morning, showed greater inhibition (96.4%). In the analysis of the cytotoxicity activity performed with larvae of Artemia salina, the essential oil extracted from the leaf collected in the rainy period in the morning, and the essential oil extracted from the flower, collected in the rainy season, presented higher toxicity. The ethanolic extracts of the leaf and the hexane extracts of the flower, both collected during the rainy season, were also more toxic. In the antimicrobial assays, flower oils collected in the rainy and dry period demonstrated greater inhibition against the bacteria Staphylococcus aureus and Bacillus cereus. The Candida albicans yeast only showed inhibition with the essential oils of the flower collected in the dry period. In the rainy season only the essential oil extracted from the leaf collected in the morning and afternoon, showed inhibition against the bacteria S. aureus and B. cereus. Regarding the dry period, the essential oil that showed the highest inhibition in B. cereus bacteria was collected in the afternoon. The hexane extracts of the flower collected in the rainy season showed greater inhibition of S. aureus and in the leaves collected in the rainy season. This extract showed greater toxicity with the bacterium Salmonella typhymurium. In the test against yeast C. albicans, the most toxic hexanic extract was that of the leaf collected in the dry period. In this work the essential oils and extracts of H. dilatata were also tested in larvicidal activities of Aedes aegypti, Anopheles darlingi and adulticides with Ae. aegypti. The essential oil with higher toxicity in the larvicidal activities of Ae. aegypti were extracted from the leaf collected in the rainy season, at night time and from the flower in the rainy season. Only the hexane extracts extracted from the flowers showed toxic activity on the larvae of Ae. aegypti. In larvae of An. darlingi, toxicity predominated with the essential oils of the flowers extracted in the dry period and in the essential oil of the leaf extracted in the same period, in the morning. In the adulticidal assays of Ae. aegypti, leaf oil collected in the rainy season at night showed a mortality of 54.4% to 91.9%, at concentrations of 62.5 μg/mL -1 and 1000 μg/mL-1 respectively in the range of 90 minutes of exposure. The knockdown effect (mosquito immobility) was observed in the test for the development of the product prototype with leaf essential oil, collected in the rainy season and in the morning. For that, the insecticidal activity against the mosquito Ae. aegypti, in 20 minutes at the start of the test, with more than 50% of mosquitoes in knockdown state. / As espécies da família Lamiaceae são conhecidas no Brasil por serem bastante aromáticas e por apresentarem propriedades bioativas como antioxidante, bactericida, fungicida e inseticida. Neste trabalho foi analisada a espécie Hyptis dilatata Benth, encontrada em áreas de cerrado no Brasil. Estudou-se os componentes químicos, a atividade biológica do óleo essencial e dos extratos de folhas e flores, considerando os períodos seco e chuvoso, de espécimes da Serra do Tepequém, em Roraima-Brasil. Os óleos essenciais foram extraídos das folhas coletadas em diferentes horários (manhã, tarde e noite) e das flores nos períodos seco e chuvoso. Em relação aos extratos hexânicos e etanólicos, oriundos das folhas e flores, estes foram extraídos de exemplares coletados nos períodos seco e chuvoso. Os resultados do perfil fitoquímico, originados a partir da análise dos extratos hexânicos e etanólicos das folhas e flores, apresentaram diferentes metabólitos secundários, sendo identificados cumarina, flavonóides, fenóis, saponinas, alcalóides, entre outros. A caracterização química dos óleos essenciais das folhas e flores mostrou como substâncias majoritárias α-pineno, β-pineno, limoneno, 3-careno, β-cariofileno, fenchona e cânfora. O óleo essencial extraído das flores, no período chuvoso e seco, inibiu a enzima acetilcolinesterase, respectivamente, 93,4% e 92,4%, enquanto que o óleo essencial extraído das folhas, no período seco e pela manhã, apresentou maior inibição (96,4%). Na análise da atividade de citotoxidade, realizadas com larvas de Artemia salina, o óleo essencial extraído da folha coletada no período chuvoso, pela manhã, e o óleo essencial extraído da flor, coletada no período chuvoso, apresentaram maior toxicidade. Apresentaram também maior toxicidade os extratos etanólicos da folha e os extratos hexânicos da flor, ambas coletadas no período chuvoso. Nos ensaios antimicrobianos, os óleos das flores coletadas no período chuvoso e seco demonstraram maior inibição contra as bactérias Staphylococcus aureus e Bacillus cereus. A levedura Candida albicans só apresentou inibição com os óleos essênciais da flor coletada no período seco. No período chuvoso, somente o óleo essencial extraído das folhas coletadas no horário da manhã e tarde, mostraram inibição contra as bactérias S. aureus e B. cereus. Em relação ao período seco, o óleo essencial que apresentou maior inibição contra bactérias B. cereus foi coletado no horário da tarde. Os extratos hexânicos da flor coletada no período chuvoso mostraram maior inibição contra S. aureus e nas folhas coletadas no período chuvoso. Este extrato demonstrou maior toxicidade com a bactéria Salmonella typhymurium. No teste contra a levedura C. albicans, o extrato mais tóxico foi o hexânico da folha coletada no período seco. Neste trabalho também foram testados os óleos essenciais e extratos de H. dilatata em atividades larvicidas de Aedes aegypti, Anopheles darlingi e teste adulticidas com Ae. aegypti. O óleo essencial com maior toxicidade nas atividades larvicidas de Ae. aegypti foram extraídos da folha coletada no período chuvoso, no horário da noite e da flor no período chuvoso. Somente os extratos hexânicos extraídos das flores apresentaram atividade tóxica sobre as larvas de Ae. aegypti. Nas larvas de An. darlingi, a toxicidade predominou com os óleos essenciais das flores extraídas no período seco e no óleo essencial da folha extraído no mesmo período, no horário da manhã. Nos ensaios adulticida de Ae. aegypti, o óleo das folhas coletadas no período chuvoso, no horário da noite, apresentou mortalidade entre 54,4% a 91,9%, nas concentrações de 62,5 μg mL-1 e 1000 μg mL-1, respectivamente, no intervalo de 90 minutos de exposição. No teste para o desenvolvimento do protótipo do produto com o óleo essencial da folha, coletada no período chuvoso e no horário da manhã, foi observada a ação do efeito knockdown (imobilidade do mosquito). Para tanto, considerou-se a atividade inseticida contra o mosquito Ae. aegypti, em 20 minutos no início do teste, com mais de 50% dos mosquitos em estado de knockdown.

As alterações das paisagens promovidas pelo homem, em razão das atividades relacionadas ao uso e ocupação do solo, representam um desafio para as atividades de controle da malária na Amazônia brasileira. Desse modo, buscou-se avaliar o sistema ecológico da malária através da construção de três eixos: desmatamento, uso do solo e diversidade de Culicidae. Esses eixos tiveram a paisagem como centro de conexão, modulados por fatores de pressão (hospedeiro humano), de risco (o vetor) e de causa (o agente infeccioso). A transmissão de patógenos, incluindo espécies de Plasmodium, ocorre na intersecção entre os nichos do hospedeiro humano, dos vetores e dos parasitos, em ambiente que permite a interconexão dos mesmos. Nesse sentido, verificou-se que cada quilômetro quadrado de área impactada pelo desmatamento, entre 2009-2015, produziu 27 novos casos de malária (r² = 0,78; F1,10 = 35,81; P <0,001) na Amazônia Legal brasileira, com uma correlação positiva altamente significativa entre o número de áreas de florestas impactadas com menos de 5 km² e a incidência da doença. Em virtude das relações indiretas com o desmatamento, foi possível verificar que o aumento da produção de soja, madeira, gado e óleo de palma no mundo apresentou alta correlação positiva significativa com a incidência de malária em países tropicais. No cenário brasileiro, a abundância de Nyssorhynchus darlingi respondeu positivamente à perda da cobertura florestal de áreas endêmicas de malária. Ao contrário, a diversidade de Culicidae diminuiu, deixando os vetores como espécies dominantes, favorecendo a taxa de picada e a capacidade de transmissão do Plasmodium. Desse modo, foi possível concluir que a incidência da doença, em áreas rurais, está fortemente associada aos padrões de uso e ocupação do solo. A estrutura da paisagem pode ser indicador de risco para a doença, em virtude das dinâmicas ecológicas do Ny. darlingi. / Changes in the landscapes caused by humans, due to the activities related to the use and occupation of the soil, represent a challenge for malaria control activities in the Brazilian Amazon. Therefore, we evaluated the ecological system of malaria in relation to the construction of three drivers: deforestation, land use and Culicidae diversity. These drivers had in common the landscape, modulated by factors of pressure (human host), risk (the vector) and necessary cause (the infectious agent). The transmission of pathogens, including species of Plasmodium, occurs at the intersection among niches of the human host, the vectors and the parasites, in an environment that allows the interconnection of these organisms. The data analyzed herein verified that every square kilometer of area impacted by deforestation between 2009-2015 produced 27 new cases of malaria (r² = 0.78, F1.10 = 35.81, P <0.001) in the Brazilian Legal Amazon, with a highly significant positive correlation between the number of forest areas with impacted less than 5 km² and the incidence of the disease. Due to the indirect relationship with deforestation, it was possible to verify that the increase in the production of soybean, wood, cattle and palm oil worldwide showed a significant positive correlation with the incidence of malaria in tropical countries. In the Brazilian scenario, the abundance of Nyssorhynchus darlingi responded positively to the loss of forest cover of endemic areas of malaria. In opposite, the Culicidae diversity decreased, leaving vectors as dominant species, favoring the biting rate and the capacity to Plasmodium transmission. Thus, it was possible to conclude that the incidence of the disease in rural areas is strongly associated with the patterns of land use and occupation. The structure of the landscape can be an indicator of risk for the disease, due to the ecological dynamics of the Ny. darlingi.

As alterações das paisagens promovidas pelo homem, em razão das atividades relacionadas ao uso e ocupação do solo, representam um desafio para as atividades de controle da malária na Amazônia brasileira. Desse modo, buscou-se avaliar o sistema ecológico da malária através da construção de três eixos: desmatamento, uso do solo e diversidade de Culicidae. Esses eixos tiveram a paisagem como centro de conexão, modulados por fatores de pressão (hospedeiro humano), de risco (o vetor) e de causa (o agente infeccioso). A transmissão de patógenos, incluindo espécies de Plasmodium, ocorre na intersecção entre os nichos do hospedeiro humano, dos vetores e dos parasitos, em ambiente que permite a interconexão dos mesmos. Nesse sentido, verificou-se que cada quilômetro quadrado de área impactada pelo desmatamento, entre 2009-2015, produziu 27 novos casos de malária (r² = 0,78; F1,10 = 35,81; P <0,001) na Amazônia Legal brasileira, com uma correlação positiva altamente significativa entre o número de áreas de florestas impactadas com menos de 5 km² e a incidência da doença. Em virtude das relações indiretas com o desmatamento, foi possível verificar que o aumento da produção de soja, madeira, gado e óleo de palma no mundo apresentou alta correlação positiva significativa com a incidência de malária em países tropicais. No cenário brasileiro, a abundância de Nyssorhynchus darlingi respondeu positivamente à perda da cobertura florestal de áreas endêmicas de malária. Ao contrário, a diversidade de Culicidae diminuiu, deixando os vetores como espécies dominantes, favorecendo a taxa de picada e a capacidade de transmissão do Plasmodium. Desse modo, foi possível concluir que a incidência da doença, em áreas rurais, está fortemente associada aos padrões de uso e ocupação do solo. A estrutura da paisagem pode ser indicador de risco para a doença, em virtude das dinâmicas ecológicas do Ny. darlingi. / Changes in the landscapes caused by humans, due to the activities related to the use and occupation of the soil, represent a challenge for malaria control activities in the Brazilian Amazon. Therefore, we evaluated the ecological system of malaria in relation to the construction of three drivers: deforestation, land use and Culicidae diversity. These drivers had in common the landscape, modulated by factors of pressure (human host), risk (the vector) and necessary cause (the infectious agent). The transmission of pathogens, including species of Plasmodium, occurs at the intersection among niches of the human host, the vectors and the parasites, in an environment that allows the interconnection of these organisms. The data analyzed herein verified that every square kilometer of area impacted by deforestation between 2009-2015 produced 27 new cases of malaria (r² = 0.78, F1.10 = 35.81, P <0.001) in the Brazilian Legal Amazon, with a highly significant positive correlation between the number of forest areas with impacted less than 5 km² and the incidence of the disease. Due to the indirect relationship with deforestation, it was possible to verify that the increase in the production of soybean, wood, cattle and palm oil worldwide showed a significant positive correlation with the incidence of malaria in tropical countries. In the Brazilian scenario, the abundance of Nyssorhynchus darlingi responded positively to the loss of forest cover of endemic areas of malaria. In opposite, the Culicidae diversity decreased, leaving vectors as dominant species, favoring the biting rate and the capacity to Plasmodium transmission. Thus, it was possible to conclude that the incidence of the disease in rural areas is strongly associated with the patterns of land use and occupation. The structure of the landscape can be an indicator of risk for the disease, due to the ecological dynamics of the Ny. darlingi.

Les modèles de distribution d’espèces ont été identifiées comme pertinents pour cartographier et caractériser la qualitéd’habitat des moustiques vecteurs du paludisme, Anopheles, afin de participer à l’estimation du risque de transmission decette maladie et à la définition de stratégies de lutte anti-vectorielle ciblées. La transmission du paludisme dépend de laprésence et de la distribution des vecteurs, qui dépendent elles-mêmes des conditions environnementales définissant laqualité des habitats écologiques des Anopheles. Cependant, dans certaines régions, les données de captures d’Anophelessont rares, et rend difficile la cartographie de leurs habitats. De plus, le recueil de ces données est très souvent soumis à desbiais d’échantillonnage.Cette thèse fournit une solution à la cartographie des vecteurs du paludisme, en considérant deux aspects très peu étudiésdans la modélisation : le faible nombre de sites de présence disponibles et l’existence d’un biais d’échantillonnage. Uneméthode originale de correction de l’effet du biais d’échantillonnage est proposée puis appliquée à des données de présencedu principal vecteur du paludisme en Guyane, Anopheles darlingi. Un modèle de distribution d’An. darlingi a ensuite étéconstruit, permettant d’obtenir une carte de qualité d’habitat en cohérence avec la connaissance des entomologistes etfournissant des performances de prédiction élevées. La méthode de correction proposée a ensuite été comparée auxméthodes existantes dans un contexte applicatif caractérisé par la rareté des données d’occurrence de l’espèce et laprésence d’un biais d’échantillonnage. Les résultats montrent que la méthode développée est adaptée aux cas où le nombrede sites de présence est faible. Cette thèse contribue, d’une part, à combler les lacunes théoriques et d’applicabilité desméthodes actuelles visant à corriger l’effet des biais d’échantillonnage et, d’autre part, à compléter la connaissance sur ladistribution spatiale et la bioécologie du principal vecteur du paludisme en Guyane française. / Species distribution models are identified as relevant to map and characterize the habitat quality of Anopheles genusmosquitoes, transmitting malaria, and thus to both participate in the estimation of the transmission risk of this disease and inthe definition of targeted vector control actions. The malaria transmission depends on the presence and distribution of thevectors, which are themselves dependent on the environmental conditions that define the quality of the ecological habitats of the Anopheles. However, in some areas, Anopheles collection data remain scarce, making it difficult to model these habitats. In addition, the collection of these data is very often subjected to significant sampling biases, due, in particular, to unequal accessibility to the entire study area. This thesis provides a solution to the mapping of malaria vectors, considering two very few studied aspects in modeling: the low number of available presence sites and the existence of a sampling bias. An original method for correcting the effect of the sampling bias is proposed and then applied to presence data of Anopheles darlingi species - the main vector of malaria in South America - in French Guiana. Then, a distribution model of An. darlingi was built to obtain a map of habitat quality consistent with entomologists’ knowledge and providing high prediction performances. The proposed correction method was then compared to existing methods in an application context characterized by the scarcity of the species occurence data and the presence of a sampling bias. The results show that the developed method is adapted to cases where the number of sites of presence is low. This thesis contributes, on the one hand, to fill theoretical and applicability lacuna of current methods intended to correct the effect of the sampling bias and, on the other hand, to supplement the knowledge on both the spatial distribution and the bio-ecology of the main malaria vector in French Guiana.

Introdução - A malária é uma das principais doenças humanas do mundo e afeta principalmente as populações pobres em regiões tropicais e subtropicais onde as condições ambientais são favoráveis tanto para a proliferação dos agentes etiológicos como dos mosquitos vetores. No Brasil, An. darlingi é considerado vetor primário de plasmódios humanos. Devido à importância médica, esse inseto tem sido objeto de campanhas de controle populacional. No entanto, não foi considerada a ocorrência de microevolução em An. darlingi e consequentemente, a possibilidade de populações diferentes apresentarem características biológicas distintas. Objetivo Verificar a presença de variabilidade genética e morfológica em populações de An. darlingi no Brasil. Metodologia - Foram analisados e comparados 26 atributos da morfologia externa de ovos de nove populações, bem como a morfometria geométrica da forma alar de espécimes de 10 localidades diferentes de Anopheles darlingi. Além disso, foram empregadas sequências do gene mitocondrial citocromo oxidase subunidade I (COI) para analisar a estrutura populacional de An. darlingi. Resultados e Conclusão Apesar dos atributos dos ovos apresentarem variação, somente as amostras de Tocantins e Pará foram diferenciadas das demais populações. As variações nas estruturas externas dos ovos são provavelmente adaptativas, com influência de fatores ambientais, como temperatura, umidade e disponibilidade de alimento para as fêmeas. A comparação da morfometria geométrica da asa demonstrou que existe maior similaridade entre as populações da costa (estados do Espírito Santo , do cerrado (estados de Goiás e Tocantins), do norte do rio Amazonas (estados do Amazonas e Amapá), e do sul do rio Amazonas (estados de Mato Grosso e Pará). As análises de COI demonstraram diferenciação das populações da costa da Mata Atlântica em relação às demais populações do Brasil central, Amazônia e Mata Atlântica do Interior. O padrão da diversidade de haplótipos de COI, os testes de neutralidade e a distribuição de diferenças pareadas indicaram recente expansão demográfica. A distância geográfica, as ecorregiões e fatores ambietais podem provavelmente limitar a dispersão da espécie, tendo alguma influência sobre a estrutura populacional de An. darlingi / Introduction Malaria is a major human disease that primarily affects poor persons in tropical and subtropical regions where environmental conditions are favorable to the proliferation of the etiological agent as well as vector mosquitoes. In Brazil, Anopheles darlingi is considered a primary vector of human plasmodia and has been the target of control campaigns because of its medical importance. However, the occurrence of microevolution in An. darlingi was not considered and, consequently, the possibility of different populations exhibit distinct biological characteristics. Objective Verify the presence of morphological and genetic variability in Brazilian Anopheles darlingi populations. Methods Twenty six attributes of external egg morphology were analyzed and compared in nine populations. Similarly, the geometric morphometry of wing shape was evaluated in ten An. darlingi localities. In addition, the mitochondrial gene cytochrome oxidase subunit I (COI) was employed to analyze the population structure of An. darlingi. Results and Conclusion - Although, egg attributes showed variation, only samples from Tocantins and Pará were differentiated from other populations. Variation in external egg structure is arguably adaptive in light of environmental factors such as temperature, humidity and food availability for females. Comparisons of wing geometric morphometry indicated that there is greater similarity between Coastal (Espírito Santo and Rio de Janeiro states) and Inner (São Paulo and Paraná states) populations of the Atlantic Forest, and differentiation of those from (Goiás and Tocantins states) and regions both north (Amazonas and Amapá states) and south (Pará and Mato Grosso states) of the Amazon Region. Cytochrome oxidase I analyses indicated differentiation between coastal Atlantic Forest populations (Espírito Santo and Rio de Janeiro states) and those from the cerrado, the Amazon and the interior Atlantic Forest. The pattern of COI haplotype diversity, neutrality tests and distribution of pairwise differences suggest a recent demographic expansion. Geographical distance, ecoregions and potentially environmental constraints may limit the species´ dispersal and influence the population structure of An. darlingi

Introdução - A malária é uma das principais doenças humanas do mundo e afeta principalmente as populações pobres em regiões tropicais e subtropicais onde as condições ambientais são favoráveis tanto para a proliferação dos agentes etiológicos como dos mosquitos vetores. No Brasil, An. darlingi é considerado vetor primário de plasmódios humanos. Devido à importância médica, esse inseto tem sido objeto de campanhas de controle populacional. No entanto, não foi considerada a ocorrência de microevolução em An. darlingi e consequentemente, a possibilidade de populações diferentes apresentarem características biológicas distintas. Objetivo Verificar a presença de variabilidade genética e morfológica em populações de An. darlingi no Brasil. Metodologia - Foram analisados e comparados 26 atributos da morfologia externa de ovos de nove populações, bem como a morfometria geométrica da forma alar de espécimes de 10 localidades diferentes de Anopheles darlingi. Além disso, foram empregadas sequências do gene mitocondrial citocromo oxidase subunidade I (COI) para analisar a estrutura populacional de An. darlingi. Resultados e Conclusão Apesar dos atributos dos ovos apresentarem variação, somente as amostras de Tocantins e Pará foram diferenciadas das demais populações. As variações nas estruturas externas dos ovos são provavelmente adaptativas, com influência de fatores ambientais, como temperatura, umidade e disponibilidade de alimento para as fêmeas. A comparação da morfometria geométrica da asa demonstrou que existe maior similaridade entre as populações da costa (estados do Espírito Santo , do cerrado (estados de Goiás e Tocantins), do norte do rio Amazonas (estados do Amazonas e Amapá), e do sul do rio Amazonas (estados de Mato Grosso e Pará). As análises de COI demonstraram diferenciação das populações da costa da Mata Atlântica em relação às demais populações do Brasil central, Amazônia e Mata Atlântica do Interior. O padrão da diversidade de haplótipos de COI, os testes de neutralidade e a distribuição de diferenças pareadas indicaram recente expansão demográfica. A distância geográfica, as ecorregiões e fatores ambietais podem provavelmente limitar a dispersão da espécie, tendo alguma influência sobre a estrutura populacional de An. darlingi / Introduction Malaria is a major human disease that primarily affects poor persons in tropical and subtropical regions where environmental conditions are favorable to the proliferation of the etiological agent as well as vector mosquitoes. In Brazil, Anopheles darlingi is considered a primary vector of human plasmodia and has been the target of control campaigns because of its medical importance. However, the occurrence of microevolution in An. darlingi was not considered and, consequently, the possibility of different populations exhibit distinct biological characteristics. Objective Verify the presence of morphological and genetic variability in Brazilian Anopheles darlingi populations. Methods Twenty six attributes of external egg morphology were analyzed and compared in nine populations. Similarly, the geometric morphometry of wing shape was evaluated in ten An. darlingi localities. In addition, the mitochondrial gene cytochrome oxidase subunit I (COI) was employed to analyze the population structure of An. darlingi. Results and Conclusion - Although, egg attributes showed variation, only samples from Tocantins and Pará were differentiated from other populations. Variation in external egg structure is arguably adaptive in light of environmental factors such as temperature, humidity and food availability for females. Comparisons of wing geometric morphometry indicated that there is greater similarity between Coastal (Espírito Santo and Rio de Janeiro states) and Inner (São Paulo and Paraná states) populations of the Atlantic Forest, and differentiation of those from (Goiás and Tocantins states) and regions both north (Amazonas and Amapá states) and south (Pará and Mato Grosso states) of the Amazon Region. Cytochrome oxidase I analyses indicated differentiation between coastal Atlantic Forest populations (Espírito Santo and Rio de Janeiro states) and those from the cerrado, the Amazon and the interior Atlantic Forest. The pattern of COI haplotype diversity, neutrality tests and distribution of pairwise differences suggest a recent demographic expansion. Geographical distance, ecoregions and potentially environmental constraints may limit the species´ dispersal and influence the population structure of An. darlingi

Notre étude s’est proposée d’analyser l’incidence du paludisme et son évolution dans le temps et dans l’espace, ainsi que de rechercher les facteurs de risque d’accès palustres chez les enfants d’un village du Moyen-Oyapock (Camopi), peuplé d’Amérindiens wayampi et émerillon, d’une part, et d’un village du Haut-Maroni (Antecume Pata), peuplé d’Amérindiens wayana, d’autre part. L’approche a été multiple avec, pour chacun des deux sites d’étude :- Une analyse de survie (modèle de Cox) à partir des accès palustres confirmés biologiquement dans une cohorte d’enfants de moins de sept ans régulièrement suivis, ainsi qu’un questionnaire de type Connaissances, Attitudes et Pratiques (CAP), puis les caractéristiques des habitats et la description de leur environnement immédiat.- Une analyse spatiale avec une classification de l’occupation du sol à partir d’images satellites SPOT 5, l’extraction de variables environnementales d’intérêt, l’étude de leur effet sur la transmission du paludisme et la mise au point d’une méthode objective de sélection d’un rayon d’observation autour des habitations pour la caractérisation de l’environnement.- Une étude de séries temporelles (ARIMA) afin de déterminer l’effet des évènements climatiques et hydrologiques sur le paludisme, aux niveaux local et plus global (El Niño).Les taux d’incidence d’accès palustres sur la période 2001-2009 se sont révélés particulièrement élevés chez les jeunes enfants, notamment à Camopi avec une moyenne de 773‰ par année. Une diminution brutale de l’incidence a eue lieu en 2007 sur le Haut-Maroni et ce phénomène est observé à Camopi depuis 2010. Une prémunition se développe assez rapidement au cours de la vie (2-3 ans), surtout contre les reviviscences à Plasmodium vivax. Les facteurs environnementaux se sont avérés être les plus nombreux et les plus fortement liés à l’incidence palustre. En effet, le dégagement des alentours du carbet de toute végétation et une certaine distance de celui-ci à la forêt sont des facteurs protecteurs. La composante géographique est également apparue essentielle à Camopi avec une incidence qui variait selon le fleuve d’habitation et en fonction de la distance au hameau principal. Les facteurs météorologiques locaux (température et niveau du fleuve) se sont également révélés être liés à l’incidence du paludisme à court terme (0-3 mois). Par ailleurs, nos résultats ont permis d’émettre un certain nombre d’hypothèses quant à la transmission et au(x) vecteur(s) local(ux), et notamment de suggérer la participation d’un vecteur autre qu’An. darlingi dans la transmission du paludisme à Camopi. Nous avons également prouvé par ce travail que la télédétection et les systèmes d’information géographique sont très prometteurs pour la prise en compte de la dimension spatiale et environnementale dans l’étude des maladies transmissibles, notamment dans les zones d’accès difficile de Guyane. / The aim of our study was to analyze the incidence of malaria in children and its evolution through time and space, as well as to search for risk factors in a village in Mid-Oyapock (Camopi), populated by Amerindians Wayampi and Emerillon, on the one hand, and a village in Upper-Maroni (Antecume Pata), populated by Amerindians Wayana, on the other hand. The approach was multiple with, for both study sites:- A survival analysis (Cox modelling) completed out of biologically confirmed malaria attacks in a cohort of children under seven, as well as a Knowledge, Attitudes, Practices and Behavior (KAPB) questionnaire, and also the characteristics of the houses and the description of their immediate environment.- A spatial analysis with a land cover classification from SPOT 5 satellite images, the extraction of environmental variables, the study of their effect on malaria transmission and the development of an objective method for picking the proper observation horizon around houses in order to characterize the environment.- A time series study (ARIMA) to determine the effect of climatic and hydrological events on malaria at local and global (El Niño) scales.The incidence rates of malaria attacks during the period 2001-2009 were particularly high among young children, especially in Camopi with an average of 773‰ by year. A sharp decline in incidence occurred in 2007 on the Upper Maroni and this phenomenon has been observed in Camopi since 2010. An acquired immunity develops quite rapidly during the life (2-3 years old), especially against P. vivax relapses. Environmental factors were found to be the most strongly associated with malaria incidence. Indeed, living in a hut cleared from the surrounding vegetation and at a larger distance from the forest are protective factors. The geographic component also appeared essential in Camopi with an incidence which varied with the river of living and with the distance from the main hamlet. The local meteorological factors (temperature and river level) also proved to be linked to malaria incidence in the short term (0-3 months). Moreover, our results have allowed issuing a number of assumptions about the transmission and the local vector(s), in particular to suggest the involvement of another vector than An. darlingi in the malaria transmission in Camopi. We also proved by this work that remote sensing and geographic information systems hold great promise for the inclusion of the spatial and environmental dimensions in the study of transmitted diseases, especially in areas of difficult access in French Guiana.