Submitted by (ana.araujo@ufrpe.br) on 2016-07-05T18:07:15Z No. of bitstreams: 1 Fabio Cavalcanti Pereira.pdf: 280467 bytes, checksum: 8086a624ad0323f2488ae7dfba5c17d2 (MD5) / Made available in DSpace on 2016-07-05T18:07:15Z (GMT). No. of bitstreams: 1 Fabio Cavalcanti Pereira.pdf: 280467 bytes, checksum: 8086a624ad0323f2488ae7dfba5c17d2 (MD5) Previous issue date: 2006-02-22 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Bancroftian filariasis is a major public health problem in the tropics with 100 millions of people infected. It is a vector born infection and it is cause by Wuchereria bancrofti a parasite that lives in the human lymphatic vessels and lymph nodes affecting all ages and gender. The basic damage of the lymphatic system is lymphangiectasia which leads to lymphatic dysfunction producing lymphedema (elephantiasis), hydroecele and fistulization syndromes (chyluria, chylocele and lymph scrotum). The aim of this investigation was to study the relationship between the risk of developing fistulation syndrome and several parameters, such a amount of fat on the diet of patients assisted at Center for Teaching, Research and Tertiary Referral for bancroftian filariasis (NEPAF - Federal University of Pernambuco) in Recife, Brazil. The present study was carried out using the generalized linear models (GLMs) for the fitting of logistic model with the statistic program S-PLUS. / A Filariose Bancroftiana, ocorre como um dos maiores problemas de saúde púbica nos trópicos e afeta cerca de 100 milhões de indivíduos. Ë transmitida por um mosquito e causada pela Wuchereria bancrofti que vive nos vasos linfáticos e linfonodos dos seres humanos de todas as idades e ambos os sexos. O substrato anátomo-patológico da doença é a linfangiectasia que leva a uma disfunção linfática produzindo o linfedema (elefantíase), a hidrocele, e as síndromes de fistulização (quilúria, quilocele e linfoescroto). O objetivo dessa investigação se foi estudar a relação entre o risco de desenvolvimento das síndromes fistulizantes e vários parâmetros, tais como a quantidade de gordura na dieta de pacientes atendidos no Núcleo de Ensino, Pesquisa e Assistência em Filariose – NEPAF, Centro de Ciências da Saúde, UFPE. O presente estudo foi realizado usando-se a teoria dos modelos lineares generalizados (MLGs) para ajustar um modelo logístico para os dados da doença usando o programa estatístico S-PLUS.

Au Vietnam, malgré les succès du gouvernement dans la lutte contre le paludisme, cette maladie persiste en zones forestières, le long des frontières internationales, en particulier avec le Cambodge, et très peu de données sont disponibles sur la filariose lymphatique de Bancroft (FLB). Aucun vaccin n'étant actuellement disponible, la lutte antivectorielle ciblant les Anopheles, moustiques qui peuvent être vecteurs d'agents du paludisme et de la FLB, reste un des éléments clés pour mieux contrôler ces deux maladies. Toutefois, une meilleure connaissance des vecteurs présents en zones endémiques et l'étude de leur capacité vectorielle s'imposaient compte tenu du manque d'information sur la transmission de ces 2 maladies dans la région centre-sud du Vietnam. La capacité vectorielle étant fortement liée à la flore bactérienne de l'estomac des moustiques, sa biodiversité a été estimée afin de mieux appréhender les familles bactériennes présentes chez les moustiques de terrain, en particulier la présence d'Enterobacteriaceae qui peut avoir une influence sur la résistance des anophèles au développement de Plasmodium, agent du paludisme. Ce champ d'investigation n'avait encore jamais été étudié chez les vecteurs du paludisme au Vietnam.Cette thèse porte sur 2 objectifs principaux : (1) l'évaluation de la capacité vectorielle des espèces d'Anopheles dans la co-transmission de Plasmodium spp. et de Wuchereria bancrofti (agent de la FLB) dans la Province de Dak Nong et de Binh Phuoc du centre-sud Vietnam (à la frontière avec le Cambodge) et (2) l'estimation de la biodiversité de la flore bactérienne de l'estomac de populations d'Anopheles du centre-sud Vietnam et l'évaluation de l'influence de certaines bactéries sur la capacité vectorielle des espèces d'Anopheles. L'identification morphologique et moléculaire des spécimens d'Anopheles collectés dans les sites d'étude nous a permis d'appréhender et de mieux connaître la faune anophélienne du centre-sud Vietnam avec la présence de 24 taxa, dont des vecteurs primaires et secondaires. Les espèces dominantes sont Anopheles dirus (48,2%), An. maculatus (19,1%) et An. minimus (9,8%), trois vecteurs majeurs du paludisme et de la FLB. Pour la première fois au Vietnam, la présence d'An. scanloni, une des 8 espèces du Complexe Dirus et vecteur du paludisme en Thaïlande, a été montrée et confirmée après son identification par techniques moléculaires et séquençage. La relation taxonomique entre An. crawfordi et An. dangi, une espèce nommée de façon informelle au Vietnam en 1987, est clarifiée grâce à une étude phylogénétique qui nous a permis d'établir qu'An. dangi est un variant morphologique d'An. crawfordi.Le taux d'infection parasitaire des anophèles a été analysé grâce à des approches PCR, en temps réel et conventionnelle, afin de déterminer la capacité vectorielle des espèces collectées. Parmi les 765 spécimens d'Anopheles étudiés, 2 spécimens dont 1 An. dirus et 1 An. pampanai, ont été trouvés infectés par P. vivax. Les taux d'infection sont de 0,26% sur l'ensemble de l'échantillon, de 0,41% pour la Province de Binh Phuoc et de 0,28% chez An. dirus et 20% chez An. pampanai. Aucun moustique n'a été trouvé infecté ni par P. falciparum, ni par P. knowlesi et ni par Wuchereria bancrofti. La flore bactérienne de l'estomac des anophèles a été analysée après application de 2 méthodes d'identification basées à la fois sur la culture et la PCR du gène 16S sur 200 abdomens d'anophèles de 11 espèces différentes. Les résultats montrent la présence de 116 genres bactériens, dont 18 genres communs, classés taxonomiquement dans 7 phyla dont Acidobacteria, Actinobacteria, Bacteroidetes, Firmicutes, Planctomycetes, Proteobacteria et Synergistetes. Le genre dominant de Dak Nong est Acinetobacter et celui de Binh Phuoc est Staphyloccocus. Acinetobacter est aussi dominant dans la communauté bactérienne de tous les spécimens d'Anopheles étudiés. / In Vietnam, despite the success of the Government in controlling malaria, the disease persists in forest areas along the international borders, especially with Cambodia, and few data are available on the Bancroftian lymphatic filariasis (BLF). No vaccine is currently available to protect from these two diseases, then vector control of Anopheles mosquitoes, that may transmit both malarial and BLF agents, remains one of the key element to control these two diseases. As there is a real lack of information on the transmission of these two diseases in south-central region of Vietnam, it was necessary to apprehend the vectors occurring in endemic areas and to study their vectorial capacity. As bacterial flora in Anopheles midgut may have a strong influence on the vectorial capacity of the mosquito, its biodiversity was estimated in order to better understand bacterial families present in mosquitoes from the field, in particular the presence of Enterobacteriaceae that can have an influence on the development of Plasmodium, agent of malaria. This field of investigation of bacteria has never studied in malaria vectors of Vietnam.This thesis focuses on two main objectives: (1) to evaluate the vectorial capacity of Anopheles species in the co-transmission of Plasmodium spp. and Wuchereria bancrofti (BLF agent) in the Provinces of Dak Nong and Binh Phuoc in south-central Vietnam (near the border with Cambodia), and (2) to estimate the biodiversity of the bacterial flora in the midgut of Anopheles populations of south-central Vietnam and evaluate the influence of certain bacteria on the vectorial capacity of Anopheles species.Morphological and molecular identification of Anopheles specimens collected in the study sites allowed us to apprehend and better understand the Anopheles fauna in south-central Vietnam composed of 24 taxa, including primary and secondary vectors. The dominant species were Anopheles dirus (48.2%), An. maculatus (19.1%) and An. minimus (9.8%), three major vectors of malaria and the BLF. For the first time, An. scanloni, one of the 8 species of Dirus Complex and malaria vector in Thailand, was collected in Vietnam and confirmed by molecular and sequencing techniques. The relationship between An. crawfordi and An. dangi, a species informally named in Vietnam in 1987, was clarified through a phylogenetic study that allows us to establish that An. dangi is a morphological variation of An. crawfordi.The parasites infection rate of Anopheles was investigated using both real-time PCR and conventional PCR to determine the vectorial capacity of the collected species. Of the 765 Anopheles specimens, 2 individuals, such as 1 An. dirus and 1 An. pampanai, were found infected by P. vivax. Then, the infection rates were of 0.26% on the total sample, 0.41% in Binh Phuoc, 0.28% for An. dirus, and 20% for An. pampanai. No mosquito was found infected by either P. falciparum, P. knowlesi or Wuchereria bancrofti.The bacterial flora in Anopheles midguts was analyzed using 2 identification methods based on culture and on 16S PCR-TTGE processed on 200 abdomens of 11 different Anopheles species. The results obtained showed the presence of 116 bacterial genera, including 18 common genera, belonging to 7 phyla such as Acidobacteria, Actinobacteria, Bacteroidetes, Firmicutes, Planctomycetes, Proteobacteria, and Synergistetes. The dominant genus in Dak Nong was Acinetobacter and Staphyloccocus in Binh Phuoc. Acinetobacter was dominant in the bacterial community of all studied Anopheles. The genus Enterobacter, which can influence the development of the Plasmodium, represented a prevalence of 1.7% of the microbiome of our specimens compared to 39.2% for the predominant genus Acinetobacter. This latter genus was significantly associated to Anopheles infected with Plasmodium vivax.

Introdução: Este trabalho tem por objetivo identificar focos de filariose linfática na amazônia ocidental brasileira e constatar a infecção natural pela W. bancrofti do mosquito vetor, o C. quinquefasciatus. O estudo abrangeu as cidades de Porto Velho (RO), Guajará Mirim (RO) e Humaitá (AM). Método: foi utilizada a técnica da gota espessa de sangue, colhida após 22h00, corada com Giemsa para avaliação humana, e a Reação em Cadeia da Polimerase (PCR) para pesquisa de DNA de W. bancrofti em mosquitos vetores. Foram analisadas e avaliadas amostras do sangue de moradores dos bairros mais antigos dessas cidades e escolares noturnas e captura de mosquitos. Resultados: O inquérito hemoscópico em moradores resultou em 935 indivíduos (54,4%) examinados de um total de 1.720 cadastrados. As pesquisas entre escolares noturnos envolveram 2.709 indivíduos (75,2%) de um total de 3.601. Todos negativos para a presença de microfilaremia. Foram coletadas 7.849 fêmeas de C. quinquefasciatus, todas apresentaram resultados negativos para DNA de W. bancrofti. / Introduction: This work is aimed to identify possible lymphatic filariasis foci in the Western Brazilian Amazonian. Porto Velho and Guajara-Mirim (Rondonia State) and Humaita (Amazonas State) were the study target area. Methods: Blood thick film method with samples collected from 10 PM to 1 AM were stained using Giemsa to evaluate human infection and PCR for W. bancrofti DNA in mosquitoes vectors. Samples from the neighborhoods of these areas and from students attending public nighttime classes in the cities referred to above were analyzed and evaluated, as well as mosquitoes captured in the houses. Results: 935 individuals (54.4%) out of a total of 1,720 individuals engaged. Sample with night students involving 2,709 individuals (75.2%) out of the total of 3,601 previously selected. No individual examined was positive for the presence of microfilariae in the blood stream. 7,849 female C.quinquefasciatus specimens were captured and after evaluated by using PCRmethod, all of them were found negative for W. bancrofti DNA.

Introdução: Este trabalho tem por objetivo identificar focos de filariose linfática na amazônia ocidental brasileira e constatar a infecção natural pela W. bancrofti do mosquito vetor, o C. quinquefasciatus. O estudo abrangeu as cidades de Porto Velho (RO), Guajará Mirim (RO) e Humaitá (AM). Método: foi utilizada a técnica da gota espessa de sangue, colhida após 22h00, corada com Giemsa para avaliação humana, e a Reação em Cadeia da Polimerase (PCR) para pesquisa de DNA de W. bancrofti em mosquitos vetores. Foram analisadas e avaliadas amostras do sangue de moradores dos bairros mais antigos dessas cidades e escolares noturnas e captura de mosquitos. Resultados: O inquérito hemoscópico em moradores resultou em 935 indivíduos (54,4%) examinados de um total de 1.720 cadastrados. As pesquisas entre escolares noturnos envolveram 2.709 indivíduos (75,2%) de um total de 3.601. Todos negativos para a presença de microfilaremia. Foram coletadas 7.849 fêmeas de C. quinquefasciatus, todas apresentaram resultados negativos para DNA de W. bancrofti. / Introduction: This work is aimed to identify possible lymphatic filariasis foci in the Western Brazilian Amazonian. Porto Velho and Guajara-Mirim (Rondonia State) and Humaita (Amazonas State) were the study target area. Methods: Blood thick film method with samples collected from 10 PM to 1 AM were stained using Giemsa to evaluate human infection and PCR for W. bancrofti DNA in mosquitoes vectors. Samples from the neighborhoods of these areas and from students attending public nighttime classes in the cities referred to above were analyzed and evaluated, as well as mosquitoes captured in the houses. Results: 935 individuals (54.4%) out of a total of 1,720 individuals engaged. Sample with night students involving 2,709 individuals (75.2%) out of the total of 3,601 previously selected. No individual examined was positive for the presence of microfilariae in the blood stream. 7,849 female C.quinquefasciatus specimens were captured and after evaluated by using PCRmethod, all of them were found negative for W. bancrofti DNA.