Dípteros (Culicidae) transmissores de arbovírus em área de proteção ambiental urbana (Parque das Dunas Natal-RN,) 2004-2006

Desenvolvido no Parque das Dunas, segunda reserva ambiental urbana do Brasil, ocupando uma área de 1.172,80 hectares, com característica de mata atlântica de dunas, situada numa faixa litorânea na região urbana do município de Natal (05° 46 S, 35º 12 W), o presente estudo, realizado durante os anos de 2004 a 2006, teve como objetivo identificar as espécies de culicídeos existentes no Parque das Dunas, capazes de transmitir arbovírus, tendo em vista que em 2004 houve uma epizootia de saguis (Callitrix jacchus), que causou grande mortandade, sem definição do agente etiológico. No ano de 2004, foram pesquisados sete pontos no interior da mata, com instalação de 20 armadilhas de ovitrampas e 20 de bambu para coleta dos imaturos. Para os adultos, durante quatro vezes por semana, foram usadas as armadilhas de Sannhon. Foram coletados 5.691 imaturos, sendo 839 Ae. aegypti, 3.184 Ae. albopictus e 1.668 Hg. leucocelaenus. A coleta dos adultos foi realizada de 2004 a 2006, etapa em que se recolheu 17.506 culicídeos adultos, sendo 17.244 Wy. bourrouli, 255 Ae. aegypti, 593 Ae. albopictus, 1.275 Hg. leucocelaenus, 294 Oc. scapularis, 05 Oc. taeniorynchus, 02 Oc. serratus e 3 Li. durhami. Para os imaturos houve correlação significativa entre Ae. aegypti e umidade relativa do ar p = 0, 049 e pluviometria p = 0,00, Ae. albopictus apresentou correlação significativa positiva com a pluviometria, enquanto Hg. leucocelaenus não apresentou nenhuma das variáveis climáticas. Para os adultos, a análise de série temporal aponta flutuação sazonal significativa para Ae. aegypti (p = 0,003); Ae. albopictus (p = 0,04); Oc. scapularis (p = 0,008 ) e Hg. leucocelaenus (p = 0,003). Uma correlação significativa negativa foi observada entre o número de Ae. albopictus coletado e a temperatura (Corr= - 0,50, p = 0,01); isto é, para cada 1°C a mais há diminuição de 7 espécimes. Este estudo teve a participação de uma equipe multidisciplinar: biólogos, entomologistas, para confirmação das espécies; técnicos de laboratório, para acompanhamento diário das larvas eclodidas das armadilhas de ovitrampas. Teve a importante colaboração de profissionais da Fundação Oswaldo Cruz FIOCRUZ/Rio de Janeiro, da Universidade de São Paulo USP para identificação do grupo Wyeomyia.

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

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Short report: Application of an alternative Aedes species (Diptera : Culicidae) surveillance method in Botucatu City, São Paulo, Brazil

One of the main problems with dengue is the control of Aedes aegypti, its major vector. In Brazil, the current control program for Ae. aegypti and Aedes albopictus populations includes larval density surveys. An interesting alternative is the use of a distinct index, the Premise Condition Index (PCI). This tool relates conditions of property, such as houses and yards, and the degree of shade with the occurrence of Aedes sp. oviposition, and is calculated as scores from 3 to 9. The lowest score indicates property in good condition and an unfavorable breeding environment, while the highest score indicates property at high risk for infestation by Aedes sp. The present study is based on the application of the PCI in an urban area of Botucatu, Brazil to confirm its effectiveness.

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)

Culture-dependent and culture-independent characterization of microorganisms associated with Aedes aegypti (Diptera: Culicidae) (L.) and dynamics of bacterial colonization in the midgut

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Caffeine effect on mortality and oviposition in successive generations of Aedes aegypti

OBJETIVO: Experimentos anteriores mostraram que a cafeína bloqueia o desenvolvimento de Aedes aegypti (Diptera, Culicidae) na fase larval, inibindo conseqüentemente a produção de adultos. O objetivo do estudo foi obter dados que pudessem sugerir desenvolvimento de resistência dos mosquitos à cafeína. MÉTODOS: Foi avaliada a produção de adultos em gerações sucessivas, a partir de ovos produzidos na geração anterior e a taxa de oviposição em cada geração, utilizando meios contendo cafeína a 200 e 500 µg/ml e água de torneira proveniente de poço artesiano como controle. Os experimentos foram conduzidos em São José do Rio Preto, entre 2002 e 2005. Nos testes estatísticos foram utilizados a análise exploratória de dados e algoritmos de alisamento. RESULTADOS: Ocorreu redução crescente da produção de adultos, nas duas concentrações, ao longo das gerações, mas apenas no experimento a 200 µg/ml os dados foram estatisticamente significantes. Quanto à oviposição, a análise dos números mostra redução crescente e acentuada na média de ovos por fêmea, no experimento tratado. CONCLUSÕES: Não houve evidência de resistência ao longo das gerações devido ao tratamento com cafeína. Os resultados encontrados podem reforçar a indicação da cafeína como uma alternativa aos principais agentes de controle do Ae. aegypti atualmente usados, contra os quais os mosquitos têm desenvolvido resistência.

Effects of caffeine and used coffee grounds on biological features of Aedes aegypti (Diptera, Culicidae) and their possible use in alternative control

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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.

Allozyme differentiation among allopatric populations of Anopheles nuneztovari (Diptera: Culicidae)

Four enzymes in six geographic populations, four Brazilian and two Colombian, of Anopheles nuneztovari were studied. There were differences among the most frequent alleles in the EST5, ACON and MDH loci in the populations from Brazil and Colombia. The α-GPD(*)C allele was encountered very frequently in the Palo/Sit population in Colombia and the α-GPD(*)B allele was found to be fixed in most of the Brazilian populations. An additional α-GPD band was found only in Palo/Sit. The considerable genetic differentiation between populations in western Colombia vs. Brazil suggests that a certain degree of reproductive isolation exists between them.

Variation of genetic expression during development, revealed by esterase patterns in Aedes aegypti (Diptera, Culicidae)

Polyacrylamide gel electrophoresis was used to analyze esterase patterns during development of Aedes aegypti from the cities of Marília and São José do Rio Preto (SJRP), Brazil. The zymograms showed a total of 23 esterase bands, 22 of which were in the specimens from Marília and 19 in those from SJRP. These esterase bands were considered to be the product of 23 alleles distributed tentatively in eight genetic loci. Most of the alleles were developmentally regulated. The larval stage expressed the greatest number of them (19 alleles, from the eight loci, in Marília; and 17 alleles, from seven loci, in SJRP). The pupal stage expressed 10 alleles from seven loci, in both populations, and the adult stage expressed 8 alleles from five and six loci in SJRP and Marília, respectively. Some alleles that were active in every stage were developmentally controlled at the level of expression (amount of product). A single allele was constitutively and highly expressed, in larvae, pupae, and adults, in both populations. Differences in esterase synthesis among stages are probably due to regulatory mechanisms acting in agreement with the requirements of a variable number of processes in which esterases are involved. The larval stage is the most active in developmental processes and shows very intense intake of food and very high mobility. These features may demand increased esterase production at that stage. Comparison of the two populations examined showed (besides the existence of alleles that they do not share) that they exhibit differences in the control of expression of other alleles. Such findings may reflect genetic differences between founders in each population, but the possibility of involvement of the intensive use of insecticides in SJRP is also discussed.

Asymptomatic carriers of Plasmodium spp. as infection source for malaria vector mosquitoes in the Brazilian Amazon

We have described the existence of asymptomatic carriers of Plasmodium vivax and Plasmodium falciparum infections in native Amazon populations. Most of them had low parasitemias, detected only by polymerase chain reaction (PCR). Because they remain symptomless and untreated, we wanted to determine whether they could infect Anopheles darlingi Root, the main Brazilian vector, and act as disease reservoirs. Fifteen adult asymptomatic patients (PCR positive only) were selected, and experimental infections of mosquitoes were performed by direct feeding and by a membrane-feeding system. Seventeen adult symptomatic patients with high parasitemias were used as controls. We found an infection rate in An. darlingi of 1.2% for the asymptomatic carriers and 22% for the symptomatic carriers. Although the asymptomatic group infected mosquitoes at a much lower rate, these patients remain infective longer than treated, symptomatic patients. Also, the prevalence of asymptomatic infections is 4 to 5 times higher than symptomatic infections among natives. These results have implications for the malaria control program in Brazil, which focuses essentially on the treatment of symptomatic patients. © 2005 Entomological Society of America.

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.

Estudo da variabilidade genética de populações de Aedes aegypti (Diptera,Culicidae), resistente e suscetíveis a inseticidas

Pós-graduação em Genética - IBILCE

Avaliação do nível de suscetibilidade de linhagens de Aedes aegypti (Díptera: Culicidae) aos inseticidas utilizados para seu controle

Pós-graduação em Saúde Coletiva - FMB

Mecanismos de resistência de aedes aegyoti L. (Diptera: Culicidae) a inseticidas

Pós-graduação em Biologia Geral e Aplicada - IBB