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Phytophagy (i.e., feeding on plant-derived materials) is an essential component of mosquito biology. Yet, it has been historically neglected as most research effort has been concentrated on host-seeking behavior and pathogen transmission. As mosquitoes are the deadliest animals on earth and because challenges, such as the rise of insecticide resistance, arise, there is an urgent need for developing effective and ecologically friendly disease vector control strategies. It is therefore important to deepen our understanding of mosquito phytophagy and, consequently, its potential to develop novel vector control methods. Particular major disease vectors are Ae. aegypti and Ae. albopictus, which are spreading rapidly through the US, in part due to climate change. Herein, we first examine the effect of temperature on Ae. aegypti sugar-feeding behavior as well as overall locomotive activity and survival, using total carbohydrate assays and actometer experiments. An optimum temperature range for mosquito activity is proposed and discussed in the context of global warming. We then observe the tentative benefit provided by city-planted ornamental flowers to Ae. aegypti and Ae. albopictus living in heavily-populated, urban areas. Mosquito sugar-feeding activity and, subsequently, sugar consumption were tested for eleven commonly-planted ornamentals. Additionally, scents were collected from the headspace of each ornamental, and volatile composition was analyzed and discussed as potential cues that could mediate mosquito-plant interactions. / Master of Science in Life Sciences / Sugar-feeding is an important, but generally neglected, aspect of mosquito biology, affecting, for example, their survival, metabolism, and ability to lay eggs. While females need blood to mature their eggs, males feed exclusively on sugar, further highlighting the importance of this food source. Mosquitoes use several cues to locate flowers to feed on, including the plant scent. Aedes aegypti and Aedes albopictus are two urban species that are invasive to the US and are the vectors of several deadly pathogens including dengue, Zika and chikungunya. It is thus to study any aspect of their biology that could lead to the development of new tools to limit their propagation. Because of the nutrients provided to mosquitoes by flowering plant species, considering the dynamic ecological relationship between human, plant, and mosquito in urban, heavily populated areas is critical. Additionally, how temperature is mediating each of these interactions is important to understand and keep in consideration. Here, we first examine the effect of temperature on sugar-feeding, activity and survival in Ae. aegypti, and discuss our results in the context of potential changes in temperature caused by climate change. We also provide some insights on the role that ornamental flower species play in urban areas in the ability of these mosquito species to thrive, by attracting mosquitoes to areas where they will have access to a high number of human blood meals.

Adult and laIval midgut trypsins from the mosquito Aedes aegypli were isolated using benzamidine affinity chromatography. The cDNA of the late trypsin induced by the blood meal was isolated using an anti-trypsin monoclonal antibody, cloned and sequenced. The 862 bp sequence codes for a 257 amino acid protein, which is presumably a trypsin precursor, since the sequence of purified mosquito trypsin begins at residue 26, immediately following an arginine residue in the precursor. The amino terminal 25 amino acids in the precursor are composed of a putative 15 amino acid signal peptide and a 10 amino acid activation peptide. The activation peptide in the mosquito is different from that of vertebrate trypsinogens and suggests that activation takes place by tryptic cleavage. The deduced amino acid sequence is homologous to that of other trypsins in those residues around the catalytic triad, and in several residues which are found only in trypsins. However, the sequence of the specificity pocket in mosquito trypsin, KESPC, differs from that found in other trypsins, KDSC. The Asp is thought to bind the basic residue of the substrate, and the Glu in the mosquito trypsin may serve the same role. The changes in trypsin protein and mRNA levels following a blood meal indicate that an important component of the regulation of trypsin synthesis is at the transcriptional level. A genomic clone of "late trypsin" was isolated, mapped, and 1.2 kb of the upstream regulatory region were sequenced. The gene has no introns within the coding region. A TAT A box consensus sequence (TAT AAA) was found at position -31 from the 5' end of the mature mRNA. A cluster of five repeat sequences homologous to the GCN4 DNA binding site was found within 200 nucleotides upstream of the cap site. GCN4 is required for derepression mediated control of general amino acid biosynthesis in response to amino acid starvation in yeast. This suggests that a similar protein might regulate expression of the late trypsin gene in the mosquito. Southern blot analysis of genomic DNA suggests that the regions flanking the late trypsin coding region are polymorphic.

Aedes togoi (Theobald) from Vancouver, B.C. may enter diapause in either the egg or the larval stage. Larval diapause occurred in the 4th instar, when immatures were maintained at 16°C or less, at photoperiods shorter than 12h light/day. The critical photoperiod was calculated as LD 10.9:13.1 at 16°C. Larvae which did not pupate after 40 days at the experimental conditions were assessed as being in diapause. Non-diapause larvae reached the pupal stage in 17-20 days at 16°C. At 14°C, 100% of the larvae diapaused at LD 10:14. Temperatures of 18°C or higher, at LD 10:14, resulted in no larval diapause. Embryonic diapause resulted when immatures, adults and eggs were maintained at temperatures ranging from 22 to 28°C, at photoperiods shorter than 15h light/day. The critical photoperiod was calculated as LD 15.3:8.7 at 25°C, and LD 14.4:9.6 at 22°C. There was no diapause when the population was maintained at 24°C LD 16:8. An experiment was conducted to determine if a simulated dawn/dusk of 90 minutes/day would increase mating success. After three days at the experimental conditions, there was no difference in mating success between populations of males and females maintained with a dawn/dusk period and those maintained at the same photoperiod where the light/dark change was abrupt. / 1986

Doenças como a dengue, chikungunya, Zika e febre amarela urbana são causadas por patógenos transmitidos pela picada de fêmeas infectadas de Ae. aegypti e Ae. albopictus. Fatores bióticos, como a competição na fase larval, e abióticos, como a variação da temperatura, provocam alterações no ciclo de desenvolvimento dessas espécies. O presente estudo avaliou os efeitos da competição intraespecífica e da temperatura na atividade locomotora de fêmeas virgens de Ae. aegypti e Ae. albopictus. Os objetivos do trabalho foram: comparar o tamanho das fêmeas de Ae. aegypti e Ae. albopictus oriundas de criadouros com baixa e alta competição intraespecífica; avaliar o efeito da competição intraespecífica na atividade locomotora de fêmeas virgens de Ae. aegypti e Ae. albopictus, sob regime de 12 horas de claro e 12 horas de escuro, à 25ºC e analisar o padrão da atividade locomotora de fêmeas virgens de Ae. aegypti e Ae. albopictus, sob regime de 12 horas de claro e 12 horas de escuro, sob diferentes temperaturas (20ºC, 25ºC e 30ºC). As colônias dos mosquitos utilizados nos experimentos são provenientes da FIOCRUZ-RJ. Para o experimento de competição, larvas de ambas as espécies foram criadas sob condições de alta (100 larvas) e baixa (20 larvas) densidades. Para isso, foram utilizadas 14 bacias preenchidas com 500 ml de água e 0,2 g de ração para peixes. Após a competição, as fêmeas oriundas de cada densidade foram transferidas para incubadoras para a realização da análise da atividade locomotora. Após a atividade, verificou-se o tamanho dos adultos oriundos de cada densidade larvária, por meio da medição das asas de cada fêmea. Para a análise da atividade locomotora sob diferentes temperaturas, as fêmeas utilizadas foram desenvolvidas em ambiente com quantidade de ovos similares. Foi utilizado o mesmo procedimento para análise da atividade do experimento anterior, porém, cada experimento ocorreu sob diferentes temperaturas: 20°C, 25°C e 30°C. Para a análise dos dados obtidos foram utilizados os testes T e ANOVA multifatorial. Para o experimento de competição, fêmeas de ambas as espécies oriundas de competição de baixa densidade foram maiores do que as oriundas de desenvolvimento sob alta densidade larvária. A atividade das fêmeas de Ae. aegypti e Ae. albopictus de ambas as densidades foi muito semelhante, sugerindo que fêmeas menores tem atividade equivalente à de fêmeas maiores. Já no experimento que avaliou a atividade das fêmeas sob faixas térmicas distintas, notou-se que, em Ae. aegypti, houve diferença significativa na atividade total sob 20°C quando comparadas a 25°C e 30°C, demonstrando que a temperatura mais baixa foi limitante para a atividade desta espécie. Houve diferenças em algumas médias avaliadas na atividade de Ae. albopictus, porém, esta espécie pareceu ter sido menos influenciada com a variação da temperatura. Análises estatísticas confirmaram que as fêmeas de Ae. aegypti respondem de forma distinta das fêmeas de Ae. albopictus quando as temperaturas foram modificadas. O presente estudo ajuda a esclarecer alguns aspectos relacionados ao comportamento dos mosquitos Ae. aegypti e Ae. albopictus, que são considerados de grande importância para a saúde pública. / Diseases such as dengue, chikungunya, Zika and urban yellow fever are caused by pathogens transmitted by the bites of infected females of Ae. aegypti and Ae. albopictus. Biotics factors essentially competition on larval stages, and abiotics factors, for instance, temperature variation, occasion adjustments on development cycle of these species. The present study evaluated the effects of intraspecific competition and temperature on locomotor activity of Ae. aegypti and Ae. albopictus. The aims of this study were: to compare the size of females Ae. aegypti and Ae. albopictus from breeding sites with low and high intraspecific competition; to evaluate the effect of intraspecific competition on locomotor activity of virgin females of Ae. aegypti and Ae. albopictus, under regime of 12 hours of light and 12 hours of dark at 25°C and to analyze the pattern of the locomotor activity of virgins females of Ae. aegypti and Ae. albopictus, under regime of 12 hours of light and 12 hours of dark, under different temperatures (20ºC, 25ºC e 30ºC). The mosquitoes colonies used on experiment are from FIOCRUZ-RJ. For the competition experiment, larvae from both species were raised under conditions of high (100 larvae) and low (20 larvae) densities. For this 14 containers filled with 500 ml of water and 0,2 g of fish food were used. After the competition, the females from each density were transferred to incubators to perform locomotor activity analysis. After the activity, the size of adult from each larval density, were verified by wings measurement of each female. For the analysis of the locomotor activity under different temperatures, the females used were developed in an environment with similar amount of eggs. The same procedure was used to the analyze the activity of the previous experiment, however, each experiment happened under different temperatures: 20°C, 25°C e 30°C. For the analysis of the obtained data were applied T tests and ANOVA multivariate. For the competition experiment, females of both species from low density of competition were longer than females from competition under high larval density. The activity of females of Ae. aegypti and Ae. albopictus from both densities was very similar, suggesting that smaller females have equivalent activity to that of larger females. In the experiment that evaluated the activity of females under different thermal bands, it was noticed that in Ae. aegypti there was significant difference in total activity under 20°C when compared with 25°C and 30°C degrees, demonstrating the lower temperature was limiting to activity for this species. There were differences in some averages evaluated in Ae. albopictus activity, however, this species seemed to have been less influenced with the temperature variation. Statistical analysis confirmed that females of Ae. aegypti respond differently than females of Ae. albopictus when the temperatures were modified. The present study helps to clarify some aspects about the behavior of mosquitoes Ae. aegypti and Ae. albopictus which are considered of great concern for public health.

Doenças como a dengue, chikungunya, Zika e febre amarela urbana são causadas por patógenos transmitidos pela picada de fêmeas infectadas de Ae. aegypti e Ae. albopictus. Fatores bióticos, como a competição na fase larval, e abióticos, como a variação da temperatura, provocam alterações no ciclo de desenvolvimento dessas espécies. O presente estudo avaliou os efeitos da competição intraespecífica e da temperatura na atividade locomotora de fêmeas virgens de Ae. aegypti e Ae. albopictus. Os objetivos do trabalho foram: comparar o tamanho das fêmeas de Ae. aegypti e Ae. albopictus oriundas de criadouros com baixa e alta competição intraespecífica; avaliar o efeito da competição intraespecífica na atividade locomotora de fêmeas virgens de Ae. aegypti e Ae. albopictus, sob regime de 12 horas de claro e 12 horas de escuro, à 25ºC e analisar o padrão da atividade locomotora de fêmeas virgens de Ae. aegypti e Ae. albopictus, sob regime de 12 horas de claro e 12 horas de escuro, sob diferentes temperaturas (20ºC, 25ºC e 30ºC). As colônias dos mosquitos utilizados nos experimentos são provenientes da FIOCRUZ-RJ. Para o experimento de competição, larvas de ambas as espécies foram criadas sob condições de alta (100 larvas) e baixa (20 larvas) densidades. Para isso, foram utilizadas 14 bacias preenchidas com 500 ml de água e 0,2 g de ração para peixes. Após a competição, as fêmeas oriundas de cada densidade foram transferidas para incubadoras para a realização da análise da atividade locomotora. Após a atividade, verificou-se o tamanho dos adultos oriundos de cada densidade larvária, por meio da medição das asas de cada fêmea. Para a análise da atividade locomotora sob diferentes temperaturas, as fêmeas utilizadas foram desenvolvidas em ambiente com quantidade de ovos similares. Foi utilizado o mesmo procedimento para análise da atividade do experimento anterior, porém, cada experimento ocorreu sob diferentes temperaturas: 20°C, 25°C e 30°C. Para a análise dos dados obtidos foram utilizados os testes T e ANOVA multifatorial. Para o experimento de competição, fêmeas de ambas as espécies oriundas de competição de baixa densidade foram maiores do que as oriundas de desenvolvimento sob alta densidade larvária. A atividade das fêmeas de Ae. aegypti e Ae. albopictus de ambas as densidades foi muito semelhante, sugerindo que fêmeas menores tem atividade equivalente à de fêmeas maiores. Já no experimento que avaliou a atividade das fêmeas sob faixas térmicas distintas, notou-se que, em Ae. aegypti, houve diferença significativa na atividade total sob 20°C quando comparadas a 25°C e 30°C, demonstrando que a temperatura mais baixa foi limitante para a atividade desta espécie. Houve diferenças em algumas médias avaliadas na atividade de Ae. albopictus, porém, esta espécie pareceu ter sido menos influenciada com a variação da temperatura. Análises estatísticas confirmaram que as fêmeas de Ae. aegypti respondem de forma distinta das fêmeas de Ae. albopictus quando as temperaturas foram modificadas. O presente estudo ajuda a esclarecer alguns aspectos relacionados ao comportamento dos mosquitos Ae. aegypti e Ae. albopictus, que são considerados de grande importância para a saúde pública. / Diseases such as dengue, chikungunya, Zika and urban yellow fever are caused by pathogens transmitted by the bites of infected females of Ae. aegypti and Ae. albopictus. Biotics factors essentially competition on larval stages, and abiotics factors, for instance, temperature variation, occasion adjustments on development cycle of these species. The present study evaluated the effects of intraspecific competition and temperature on locomotor activity of Ae. aegypti and Ae. albopictus. The aims of this study were: to compare the size of females Ae. aegypti and Ae. albopictus from breeding sites with low and high intraspecific competition; to evaluate the effect of intraspecific competition on locomotor activity of virgin females of Ae. aegypti and Ae. albopictus, under regime of 12 hours of light and 12 hours of dark at 25°C and to analyze the pattern of the locomotor activity of virgins females of Ae. aegypti and Ae. albopictus, under regime of 12 hours of light and 12 hours of dark, under different temperatures (20ºC, 25ºC e 30ºC). The mosquitoes colonies used on experiment are from FIOCRUZ-RJ. For the competition experiment, larvae from both species were raised under conditions of high (100 larvae) and low (20 larvae) densities. For this 14 containers filled with 500 ml of water and 0,2 g of fish food were used. After the competition, the females from each density were transferred to incubators to perform locomotor activity analysis. After the activity, the size of adult from each larval density, were verified by wings measurement of each female. For the analysis of the locomotor activity under different temperatures, the females used were developed in an environment with similar amount of eggs. The same procedure was used to the analyze the activity of the previous experiment, however, each experiment happened under different temperatures: 20°C, 25°C e 30°C. For the analysis of the obtained data were applied T tests and ANOVA multivariate. For the competition experiment, females of both species from low density of competition were longer than females from competition under high larval density. The activity of females of Ae. aegypti and Ae. albopictus from both densities was very similar, suggesting that smaller females have equivalent activity to that of larger females. In the experiment that evaluated the activity of females under different thermal bands, it was noticed that in Ae. aegypti there was significant difference in total activity under 20°C when compared with 25°C and 30°C degrees, demonstrating the lower temperature was limiting to activity for this species. There were differences in some averages evaluated in Ae. albopictus activity, however, this species seemed to have been less influenced with the temperature variation. Statistical analysis confirmed that females of Ae. aegypti respond differently than females of Ae. albopictus when the temperatures were modified. The present study helps to clarify some aspects about the behavior of mosquitoes Ae. aegypti and Ae. albopictus which are considered of great concern for public health.

Submitted by Gizele Lima (gizele.lima@inpa.gov.br) on 2016-11-23T12:52:52Z No. of bitstreams: 2 Juliana Bruning Azevedo.pdf: 2565810 bytes, checksum: 1751c9233813fc98d009c5515f8a7e46 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-11-23T12:52:52Z (GMT). No. of bitstreams: 2 Juliana Bruning Azevedo.pdf: 2565810 bytes, checksum: 1751c9233813fc98d009c5515f8a7e46 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2015-04-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The Aedes aegypti is considered the main vector of dengue virus, urban yellow fever and Chikungunya fever. Insects are susceptible to changes in environmental factors such as rainfall and temperature. Responsible for regulating population size and aspects of mosquito biology, such as larval growth, development time, body size, longevity, fertility and blood supply. The Intergovernmental Panel on Climate Change (IPCC) predicts an increase of 2° to 4° C in average global temperature over the next century. These climatic changes may result in significant changes in landscapes and ecological patterns of infectious diseases, and interfere directly in the development and behavior of A. aegypti. This study evaluated different aspects of vector biology as amount amount of ingested blood, fertility, egg laying and hatching of eggs and longevity, climate change scenarios, planned for the year 2100, intermediate and extreme scenarios. Among the predicted variables to undergo changes due to climate change, the temperature is primarily responsible for changing the biology and vector behavior. Concentrations of CO2, even three times the current concentration does not significantly affect the results obtained in this study. The amount of blood ingested by females raised in higher temperature was higher compared to females who lived at lower temperatures, longevity is low at temperatures above 32° C, living approximately 40 days and no longevity difference between sex. The fertility is greatly affected by temperature, being reduced by half in higher temperatures. The oviposition is impaired in temperatures above 32° C, but this effect was not observed in the outbreak. Mathematical models project an increase in the number of dengue cases and geographic expansion in vector distribution. However, our data indicate that, in future scenarios of climate change, some aspects of the biology of A. aegypti will be affected. With the above information, it was possible to better understand the biology of the vector and taking also into account climate change, serving as a basis for other studies. / O Aedes aegyptié considerado o principal vetor do vírus da Dengue, Febre amarela urbana e da Febre Chikungunya. São insetos suscetíveis à variação de fatores ambientaiscomo, quantidade de chuva e temperatura. Responsáveis porregular o tamanho populacional e aspectos da biologia do mosquito, como o crescimento larval, tempo de desenvolvimento, tamanho corporal,longevidade, fecundidade e alimentação sanguínea. O Painel Intergovernamental sobre Mudança do Clima (IPCC) prevê um aumento de 2° a 4° C na temperatura média global, durante o próximo século. Essas alterações climaticas poderão resultar em mudanças significativas nas paisagens e nos padrões ecológicos das doenças infecciosas, e interferir diretamente no desenvolvimento e comportamento do A. aegypti. Neste trabalho foram avaliados diferentes aspectos da biologia do vetor como, ingestão de sague, fecundidade, oviposição e eclosão de ovos e longevidade, sobre cenários de mudanças climáticas, previstas para o ano de 2100, cenários intermediários e extremos.Dentre as variáveis preditas para sofrem alterações devido a mudanças do clima, a temperatura é principal responsável por alterar a biologia e o comportamento do vetor. As concentrações de CO2, mesmo três vezes maior que a concentração atual, não influenciou significativamente os resultados obtidos neste estudo.A quantidade de sangue ingerida por fêmeas criadas em temperatura mais elevada foi maior quando comparadas com as fêmeas que viviam em temperaturas menores, a longevidade é menor em temperaturas superiores a 32°C, vivendo aproximadamente 40 dias e não houve diferença de longevidade entre os sexos. A fecundidade é extremamente afetada pela temperatura, sendo reduzida pela metade em temperaturas maiores. A oviposição é prejudicada em temperaturas superiores a 32°C, porém esse efeito não foi observado na eclosão.Modelos matemáticos projetam um aumento no número de casos de dengue, além de uma expansão geográfica na distribuição do vetor. No entanto, nossos dados indicam que, em cenários futuros de mudanças climáticas, alguns aspectos da biologia do A. aegypti serão afetados. Com as informações acima mencionadas, foi possível conhecer melhor a biologia do vetor e levando também em consideração as mudanças climáticas, servindo como base para outros estudos.

Muitos trabalhos são desenvolvidos para identificar substâncias ou microrganismos capazes de controlar a população de insetos importantes para a saúde pública, sendo a identificação de lesões histológicas um critério para compreender o mecanismo de ação e eficácia do produto. As análises histopatológicas são baseadas na comparação entre tecidos de indivíduos controle e indivíduos expostos. Entretanto, são raros os trabalhos que referenciam o estado dos tecidos sadios. Neste trabalho caracterizou-se morfo-histoquimicamente os principais tecidos sadios do sistema digestório de larvas em 3º a 4º ínstar de Aedes aegypti, mostrando diversas imagens das variações que podem ser esperadas, além de apresentar as técnicas de processamento. Para isto, um total de trinta larvas foram fixadas, desidratadas e incluídas em historesina. Cortes foram realizados e corados com HE para morfologia e azul do Nilo, azul bromofenol, azul de toluidina e PAS para a caracterização histoquímica. Como resultados, o intestino das larvas se mostrou um tubo reto, formado por uma camada simples de células cúbicas, moderadamente acidófilas, com núcleo central e uma borda em escova voltada para o lúmen. Foram registradas variações celulares, uma vez que as células epiteliais também são secretivas das enzimas digestivas. Cortes sequenciais foram importantes para definir a morfologia, uma vez que o ângulo e altura de corte podem influenciar na sua aparência. O PAS revelou a presença de muitos grânulos de glicogênio por todos os tecidos, inclusive nas vesículas secretivas. Estas ainda possuem muitas proteínas e foram coradas fracamente para lipídios. Não foram registrados eventos de metacromasia na cor vermelha com o azul de toluidina, entretanto os tecidos parecem se corar diferentemente do tegumento. / Many surveys are designed to identify substances or microorganisms capable of controlling the population of insects that are important to public health, and the identification of resulting histological injures is a criterion for understanding the mechanism of action and the efficacy of the product. The histopathological analyses are based on the comparison between tissues of control subjects and exposed subjects, but there are rare studies that mention the state of healthy tissues. This work characterized the main morpho-histochemically healthy tissues of the digestive system of larvae in the 3rd and the 4th instar of Aedes aegypti, showing several images of the variations that can be expected, as well as displaying processing techniques. For this, a total of thirty larvae were pinned, dehydrated and embedded in historesin. Sections were made and stained with hematoxylin for morphology and Blue Nile, bromophenol blue, toluidine blue and PAS staining for the immunohistochemical characterization. As a result, the intestines of the larvae showed an upright tube formed by a single layer of cuboidal cells, moderately acidophilic, with a central nucleus and a brush border facing the lumen. Cellular changes were recorded, since epithelial cells also secrete digestive enzymes. Sequential slices were important in defining the morphology, since the angle and height of the cut may affect its appearance. The PAS revealed the presence of many glycogen granules in all tissues, including the secretory vesicles. These still have protein and were weakly colored to lipids. There were no metachromasia events in red with toluidine blue, though the tissues seem to blush differently from the integument.

Made available in DSpace on 2015-04-22T22:14:01Z (GMT). No. of bitstreams: 1 Ilia Gilmara Carvalho.pdf: 530450 bytes, checksum: 50f84ac4bf26a69a84cf2a4b2a031978 (MD5) Previous issue date: 2008-12-31 / Fundação de Amparo à Pesquisa do Estado do Amazonas / The study had for objective the detection and typing of the dengue virus, in the vectors Aedes aegypti and Aedes albopictus through the Polimerase Chain Reaction (PCR) using specific primers. During the period of December of 2005 to December of 2006 8984 mosquitoes were collected, of these 827 they were Aedes, being 819 Ae. aegypti (414 females and 405 males) and 8 Ae. albopictus (7 females and 1 male), in 46 neighborhoods of the city of Manaus including all of the geographical areas of the city. The females were grouped in pools from 1 to 10 mosquitoes for microtubes, in agreement with the gender, it dates from collection, neighborhood and stocked to -70C in the same day of the collection, totaling 143 pools (138 of Ae. aegypti and 5 of Ae. albopictus). Those pools were macerated in PBS containing albumin, and a bracket of the softened was removed for inoculation in cell culture, and other bracket was used to accomplish the extraction of the viral RNA. Once extracted, RNA was submitted the reaction of reverse transcription following by the chain reaction of the polimerase (RT-PCR). The positive samples in RT-PCR were submitted the reaction of semi nested PCR and the products of this reaction were resolved in agarose gel to 1,5% with etídio bromide. Of the 143 analyzed pools, 114 (111 of Ae. aegypti and 3 of Ae. albopictus) they were positive for DENV 3, just a pool was shown positive for two sorotipos, DENV 1 and DENV 3. The prevalence pools Ae. aegypti infected with DENV 3 in the city of Manaus was of 53%, being larger in the areas Center-west (70%), South (60%) and West (53%), and smaller in the area East (23%). This is the first report of the detection of DENV 3 in field-caught Ae. albopictus naturally infected in Brazil. The monitoring of the circulation viral in mosquitoes with the use of the technique of RT-PCR allows the previous knowledge of the levels of spread viral in certain areas contributing to determine the time and place to apply the prevention measures and control. / O estudo teve por objetivo a detecção e tipagem do vírus dengue, nos vetores Aedes aegypti e Aedes albopictus por meio da Reação em Cadeia da Polimerase (PCR) utilizando primers específicos. Durante o período de Dezembro de 2005 a Dezembro de 2006 foram coletados 8984 mosquitos, destes 827 eram Aedes, sendo 819 Ae. aegypti (414 fêmeas e 405 machos) e 8 Ae. albopictus (7 fêmeas e 1 macho), em 46 bairros da cidade de Manaus abrangendo todas as zonas geográficas da cidade. As fêmeas de Ae. aegypti e Ae. albopictus foram agrupadas em pools de 1 a 10 mosquitos por microtubos, de acordo com o gênero, data de coleta, bairro e estocado a -700C no mesmo dia da coleta, totalizando 143 pools (138 de Ae. aegypti e 5 de Ae. albopictus). Esses pools foram macerados em PBS contendo albumina, e uma alíquota do macerado foi retirada para inoculação em cultura de célula, e outra alíquota foi utilizada para realizar a extração do RNA viral. Uma vez extraído, o RNA foi submetido a reação de transcrição reversa seguida da reação em cadeia da polimerase (RT-PCR). As amostras positivas na RT-PCR foram submetidas a reação de Semi Nested PCR e os produtos desta reação foram resolvidos em gel de agarose a 1,5% corado com brometo de etídio. Dos 143 pools analisados, 114 (111 de Ae. aegypti e 3 de Ae. albopictus) foram positivos para DENV 3, apenas um pool mostrou-se positivo para dois sorotipos, DENV 1 e DENV 3. A prevalência de mosquitos Ae. aegypti infectados com DENV 3 na cidade de Manaus foi de 80%. Em todas as zonas a prevalência foi maior que 53%, sendo maior nas zonas Centro-oeste (70%), Sul (60%) e Oeste (53%), e menor na zona Leste (23%). Este foi o primeiro achado no Brasil da identificação do DENV 3 em amostras de Ae. albopictus , coletados em campo na forma adulta. O monitoramento da circulação viral em mosquitos com o uso da técnica de RT-PCR permite o conhecimento prévio dos níveis de disseminação viral em determinadas áreas contribuindo para determinar a época e local para aplicar as medidas de prevenção e controle.

Submitted by Gizele Lima (gizele.lima@inpa.gov.br) on 2018-03-07T19:05:24Z No. of bitstreams: 2 Tese JOELMA 25 01 18-1.pdf: 2217088 bytes, checksum: 4076fe5d3e54507c6b7c0ba6a34ae6e0 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-03-07T19:05:24Z (GMT). No. of bitstreams: 2 Tese JOELMA 25 01 18-1.pdf: 2217088 bytes, checksum: 4076fe5d3e54507c6b7c0ba6a34ae6e0 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-12-08 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The diversity of Bacillus thuringiensis obtained from three Brazilian biomes with pathogenic action for Ae. aegypti, An. darlingi and Cx. quinquefasciatus. Aspects as characterization of the active strains and the importance of the cyt1Aa gene also were analyzed. / Estudou-se a diversidade de Bacillus thuringiensis obtidos de três biomas brasileiros com ação patogênica para larvas de Ae. aegypti, An. darlingi e Cx. quinquefasciatus. Aspectos como caracterização molecular das linhagens ativas e a importância do gene cyt1Aa também foram analisados.