Spatial distribution of arboviral mosquito vectors (Diptera, Culicidae) in Vale do Ribeira in the South-eastern Brazilian Atlantic Forest

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Sugar digestion in mosquitoes: Identification and characterization of three midgut alpha-glucosidases of the neo-tropical malaria vector Anopheles aquasalis (Diptera : Culicidae)

Dietary carbohydrates provide an important source of energy for flight, and contribute to longevity and fecundity of mosquitoes. The most common sugar mosquitoes ingest is sucrose, and digestion of this substance is carried out mainly by alpha-glucosidases. In the current work, we tested the efficiency of sucrose on Anopheles aquasalis female diet. The best longevity (days) was reached when sugar was available in the diet, whereas most only blood fed females were dead 6 days after emergence. Three alpha-glucosidase isoforms were detected in the adult female midgut, named alpha Glu1, alpha Glu2 and alpha Glu3. These are acidic alpha-glucosidases with optima pH around pH 5.5. alpha Glu1 and alpha Glu2 are present in both secreted and membrane-bound forms, whereas alpha Glu3 only in anchored to membranes. The alpha-glucosidase activity is concentrated mainly in the posterior midgut (70%), both in non-fed or 10% sucrose fed females. The single form of these a-glucosidases seemed to be similar to 70 kDa polypeptides, although alpha Glu2 is presented in >= 600 kDa self-aggregates. K, values of alpha Glu1, alpha Glu2 and alpha Glu3 differed significantly from each other, supporting the statement that three alpha-glucosidases are produced in the female midgut. Together, all data suggest that sugar is an essential component of A. aquasalis female diet. In addition, alpha-glucosidases are synthesized in the same place where sucrose is digested and absorbed, the midgut. (c) 2007 Elsevier B.V. All rights reserved.

Genetic variability in geographical populations of Aedes aegypti (Diptera, Culicidae) in Brazil elucidated by molecular markers

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Spatial evaluation of larvae of Culicidae (Diptera) from different breeding sites: application of a geospatial method and implications for vector control

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Chitinolytic activities in the gut of Aedes aegypti (Diptera : Culicidae) larvae and their role in digestion of chitin-rich structures

Mosquito larvae are believed to be capable of digesting chitin, an insoluble polysaccharide of N-acetylglucosamine, for their nutritional benefit. Studies based on physiological and biochemical assays were conducted in order to detect the presence of chitinase activities in the gut of the detritus-feeding Aedes aegypti larvae. Larvae placed for 24 h in suspensions of chitin azure were able to digest the ingested chitin. Semi-denaturing PAGE using glycol chitin and two fluorogenic substrate analogues showed the presence of two distinct chitinase activities: an endochitinase that catalyzed the hydrolysis of chitin and an endochitinase that cleaved the short substrates [4MU(GlcNAc)(3)] and [4MU(GlcNAc)(2)] that hydrolyzed the chitobioside [4MU(GlcNAc)(2)]. The endochitinase had an extremely broad pH-activity against glycol chitin and chitin azure, pH ranging from 4.0 to 10.0. When the substrate [4MU(GlcNAc)(3)] was used, two activities were observed at pH ranges 4.0-6.0 and 8.0-10.0. Chitinase activity against [4MU(GlcNAc)(3)] was detected throughout the gut with the highest specific activity in the hindgut. The pH of the gut contents was determined by observing color changes in gut after feeding the larvae with color indicator dyes. It was observed a correlation between the pH observed in the gut of feeding larvae (pH 10-6.0) and the optimum pH for gut chitinase activities. In this work, we report that gut chitinases may be involved in the digestion of chitin-containing structures and also in the partial degradation of the chitinous peritrophic matrix in the hindgut. (C) 2003 Elsevier B.V. All rights reserved.

Genetic variation along time in a brazilian population of Aedes aegypti (Diptera : Culicidae), detected by changes in the esterase patterns

Aedes aegypti from the Brazilian cities of Sao Jose do Rio Preto (SJ) and Goiania (GO) were analyzed as to their esterase patterns and the results were compared with data obtained about 5 years before for SJ population. Esterase bands not detected in the previous study were now observed in mosquitoes from both SJ and GO populations, being the last considered a population resistant to insecticides. Other similarities between SJ and GO populations in this study, and some differences in comparison with the previous data on SJ were observed, involving, in addition to changes in band type, changes in frequency of mosquitoes expressing them and differential gene activation during development. As it is generally true for genetic features, changes in the esterase patterns are expected to be the result of factors such as selection by environmental conditions and genetic drift. In the present case, continuous use of insecticides aiming mosquito population size control in SJ by sanitary authorities could be involved in the observed changes. Changed esterases were classified as carboxylesterases and cholinesterases, which are enzymes already shown to take part in the development of resistance in several organisms. In addition, data obtained in the elapsed time by authorities responsible for the mosquito control has shown increasing insecticide resistance of SJ population mosquitoes parallel to increase in the total amount of esterases, reinforcing the mentioned possibility.

Culicidae (Diptera) em área de barragem em Santa Catarina e no Rio Grande do Sul

The Culicidae composition of the Barra Grande Lake Situated between the Municipalities of Esmeralda (Rio Grande do Sul State) and Anita Garibaldi (Santa Catarina State) was assessed by monthly samplings. Twenty-four species were identified from a total of 1, 185 specimens (74.7% as adults and 25.3% as immatures), with Aedes fluviatilis Lutz as the most frequent species. Several species are new records, and some of them are of public health interest. It is suggested that local environmental changes may alter the relationship between humans and vector mosquitoes.

First isolation of microorganisms from the gut diverticulum of Aedes aegypti (Diptera: Culicidae): New perspectives for an insect-bacteria association

We show for the first time that the ventral diverticulum of the mosquito gut (impermeable sugar storage organ) harbors microorganisms. The gut diverticulum from newly emerged and non-fed Aedes aegypti was dissected under aseptic conditions, homogenized and plated on BHI medium. Microbial isolates were identified by sequencing of 16S rDNA for bacteria and 28S rDNA for yeast. A direct DNA extraction from Ae. aegypti gut diverticulum was also performed. The bacterial isolates were: Bacillus sp., Bacillus subtilis and Serratia sp. The latter was the predominant bacteria found in our isolations. The yeast species identified was Pichia caribbica.

Efeito da cafeína no desenvolvimento de Aedes aegypti (Diptera: Culicidae): o significado biológico das alterações do padrão de síntese de esterases

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Entomological surveillance, spatial distribution, and diversity of culicidae (diptera) immatures in a rural area of the atlantic forest biome, state of sao paulo, brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Efeito de temperaturas alternadas sobre desenvolvimento de Aedes (Stegomyia) aegyoti (L.) (Diptera: Culicidae)

Aedes (Stegomyia) aegypti is a species of mosquitoes known to be the vector of diseases such as dengue and yellow fever, and a better understanding of aspects of their biology can help in the establishment of control strategies for the same. Several previous studies showed that temperature significantly affects the development of immature stages of insects. In general, higher temperatures (up to a threshold) accelerate the development of insects, and lower temperature retards the same. This rule also applies to mosquitoes, including Ae. aegypti. But not still know the effects of daily variation of temperature on the developmental stages of mosquitoes. And this detail is very important, since in natural breeding or artificial, The mosquitoes usually face temperature variations over a single day, which should interfere with its development until the emergence of the adult forms. For this reason, the objective of this study is to analyze the effect of alternating temperatures on the development of Ae. aegypti. To conduct the study, adults were collected active in the neighborhood Bela Vista Campus of UNESP - Rio Claro, SP, using a sweep net or using ovitraps for immatures, and the active search for breeding. Individuals collected were kept under experimental conditions in the laboratory. The adult samples were identified to species level, were considered for the experiments, only samples of Ae. aegypti. The insects were housed in plastic cages, suitable for creating flies. These were fed with sugar solution and blood meal on alternate days. The eggs obtained were used in the experiment with four different temperature regimes. The data collected were analyzed by evaluating whether the different treatments influenced the development of immature to adult, performing the Kruskal-Wallis test and the statistical software BioEstat. Statistical analysis of the sex ratio... (Complete abstract click electronic access below)

Avaliação dos componentes de óleos essenciais 1,8-cineol, β-cariofileno e α-humuleno como possíveis repelentes para Aedes (Stegomyia) aegypti (Diptera: Culicidae)

Aedes aegypti, mosquito da família Culicidae responsável pela transmissão dos vírus da dengue, da febre amarela urbana e da febre chikungunya, apresenta grande proliferação em áreas urbanas das regiões tropicais e subtropicais, além de representar um fator de incômodo por causar alergias devido à inoculação dos componentes salivares durante o processo do repasto sanguíneo. Como forma de evitar os problemas causados por eles, o repelente mais usado e registrado comercialmente desde 1957 é o DEET (N,N-dietil-3-metilbenzamida ou N,N-dietil-meta-toluamida), que apesar de apresentar eficácia comprovada possui determinada toxicidade quando utilizado por crianças e por mulheres grávidas ou lactantes. Atualmente, grande parte dos estudos sobre novas substâncias repelentes envolvem óleos essenciais de plantas, principalmente por serem atóxicos, biodegradáveis, possuírem um preço mais acessível e uma ampla atividade contra diferentes espécies de mosquitos. Os terpenos 1,8-cineol, β-cariofileno e α-humuleno podem ser encontrados em óleos essenciais de uma grande variedade de plantas e apesar de dados da literatura mostrarem uma eficácia dessas substâncias como repelentes para outras espécies de insetos, até o presente momento não existem estudos que relacionam porcentagem de repelência ao longo do tempo para avaliar o potencial de repelência das mesmas para culicídeos.O presente estudo avaliou tais substâncias como possíveis repelentes para A. aegypti, utilizando o DEET como controle positivo. Para avaliar a eficácia do β-cariofileno foi utilizada a BG-cage, uma gaiola desenvolvida especificamente para testes de repelência onde o voluntário expõe um braço sob uma abertura coberta por uma tela onde os mosquitos pousam, mas não picam. Os testes foram repetidos a cada 30 minutos após a aplicação do produto por um tempo máximo de duas horas (seguindo a recomendação de reaplicação do produto comercial ...

Atividade tóxica de isolados de Bacillus Thuringiensis a larvas de Aedes Aegypti (l.) (diptera: culicidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Mitochondrial DNA polymorphism and heteroplasmy in populations of Aedes aegypti in Brazil

The tropical mosquito Aedes aegypti (Diptera: Culicidae) is the most important domestic vector of urban yellow fever and dengue viruses. Ae. aegypti originated from Africa and was probably introduced into Brazil during the colonial period through embarkations, and dengue epidemics soon followed. Genetic analysis of 12 Ae. aegypti populations from five states in Brazil was conducted based on two mitochondrial DNA fragments: cytochrome oxidase I and NADH dehydrogenase subunit 4. Analyses comparing individual haplotypes indicated the existence of two well-defined clades, probably representing two mitochondrial lineages. Analysis of molecular variance showed significant variability in genetic structure among collections within groups. Mantel regression analysis showed a correlation between genetic and geographic distances, mainly because of northern and northeastern populations, in comparison with those in the southeast. The population from Santos, the largest port in Brazil, showed the greatest diversity, with 10 unique haplotypes, an indication of recent introductions that have not yet spread to other Brazilian cities. Different mitochondrial DNA sequences were found in three specimens, indicating the presence of heteroplasmy.

Seasonal population dynamics and the genetic structure of the mosquito vector Aedes aegypti in São Paulo, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)