Biotin/Avidin sandwich enzyme-linked immunosorbent assay for Culicidae mosquito blood meal identification

The knowledge of mosquitoes Culicidae host feeding patterns is basic to understand the roles of different species and to indicate their importance in the epidemiology of arthropod-borne diseases. A laboratory assay was developed aiming at standardizing the biotin-avidin sandwich enzyme-linked immunosorbent assay, which was unprecedented for mosquito blood meal identification. The enzyme-linked immunosorbent assay (ELISA) activity was evaluated by the detection of titers on each sample of the 28 blood-fed Culex quinquefasciatus. In light of the high sensitivity that the technique permits, by means of small quantities of specific antibodies commercially provided and phosphatase substrate which reinforces additional dilutions, human and rat blood meals were readily identified in all laboratory-raised Culex quinquefasciatus tested. The assay was effective to detect human blood meal dilutions up to 1:4,096, which enables the technique to be applied in field studies. Additionally, the present results indicate a significant difference between the detection patterns recorded from human blood meal which corroborate the results of host feeding patternsSecretaria de Vigilância em Saúde/Ministério da Saúd

Duration of larval and pupal development stages of Aedes albopictus in natural and artificial containers

Aedes albopictus were reared in different containers: a tree hole, a bamboo stump and an auto tire. The total times from egg hatching to adult emergence were of 19.6,27.3 and 37.5 days, respectively, according to the container. The first, second and third-instar larvae presented growth periods with highly similar durations. The fourth-instar larvae was longer than the others stages. The pupation time was longer than the fourth-instar larvae growth period. The temperature of the breeding sites studied, which was of 18&deg; C to 22&deg; C on average, was also taken into consideration. The mortality of the immature stages was analysed and compared as between the experimental groups; it was lower in the natural containers than in the discarded tire. The average wing length of adult females emerging from tree hole was significantly larger (p < 0.05) than that of those emerging from the tire.Ae. albopictus foi criado em oco de árvore, internódio de bambu e pneu de carro descartável, sob condições de campo. O resultado obtido a partir de larvas recém-nascidas até alado foi de 19,6,27,3 e 37,5 dias. As larvas dos três primeiros estádios tiveram tempo de crescimento similar, não obstante tratar-se de diferentes micro-habitats estudados. Contudo, larvas de 4º estádio apresentaram tempo de duração mais longo e crescente do oco de árvore para o pneu. A temperatura média dos três micro-habitats variou de 18&deg; Ca 22&deg; C. O resultado da taxa de mortalidade encontrada foi menor para larva e pupa do oco de árvore e maior para o pneu. A medida do comprimento médio de asa das fêmeas emergidas do oco de árvore foi maior em relação àquela do pneu

Aspectos epidemiológicos e ecológicos relacionados à malária na área de influência do lago da Represa de Porto Primavera, região oeste do Estado de São Paulo, Brasil

A study was carried out in the area of influence of the Porto Primavera Hydroelectric Power Station, in western São Paulo State, to investigate ecological and epidemiological aspects of malaria in the area and monitor the profile of the anopheline populations following the environmental changes brought about by the construction of the lake. Mosquitoes captured were analyzed by standardized indicator species analysis (ISA) before and during different flooding phases (253 m and 257 m elevations). The local human population was studied by means of parasitological (thin/thick blood smears), molecular (PCR) and serological tests. Serological tests consisted of Enzyme Linked Immunosorbent Assay (ELISA) with synthetic peptides of the circumsporozoite protein (CSP) from classic Plasmodium vivax, P. vivax variants (VK247 and "vivax-like"), P. malariae and P. falciparum and Indirect Immunofluorescence Assay (IFA) with asexual forms of P. vivax, P. malariae and P. falciparum. The results of the entomological survey indicated that, although the Anopheles darlingi population increased after the flooding, the population density remained very low. No malaria, parasite infection or DNA was detected in the inhabitants of the study area. However, there was a low frequency of antibodies against asexual forms and a significant prevalence of antibodies against P. vivax, P. vivax variants, P. falciparum and P. malariae; the presence of these antibodies may result from recent or less recent contact with human or simian Plasmodium (a parallel study in the same area revealed the existence of a sylvatic cycle). Nevertheless, these results suggest that, as in other places where malaria is present and potential vectors circulate, the local epidemiological conditions observed could potentially support the transmission of malaria in Porto Primavera Lake if infected individuals are introduced in sufficient numbers. Further studies are required to elucidate the phenomena described in this paper.Foi realizada pesquisa na área de influência do lago da Usina Hidrelétrica de Porto Primavera, região oeste do Estado de São Paulo, para estudar aspectos ecológicos e epidemiológicos da malária na localidade e acompanhar o perfil das populações de anofelinos frente às mudanças decorrentes do impacto ambiental pela formação do lago. Mosquitos capturados foram analisados pelo Índice de Abundância de Espécies Padronizado (IAEP), antes e durante o enchimento do reservatório (cotas 253 e 257 m). A população humana local foi estudada por meio de teste parasitológico (gota espessa e esfregaço sangüíneo), testes moleculares (PCR) e testes sorológicos. A sorologia consistiu na reação de ELISA com peptídeos sintéticos correspondentes à porção repetitiva da proteína circumsporozoíta (CSP) de Plasmodium vivax clássico, e suas variantes VK247 e "vivax-like", P. malariae e P. falciparum; e reação de Imunofluorescência Indireta (RIFI) com formas assexuadas de P. vivax, P. malariae e P. falciparum. Os resultados do estudo entomológico indicaram que, embora a população de Anopheles darlingi tenha aumentando após o enchimento, permaneceu em baixa densidade. Não foi detectada malária nem a presença de parasitos ou de DNA parasitário nos habitantes estudados. No entanto, foi observada baixa freqüência de anticorpos contra formas assexuadas e significativa prevalência de anticorpos contra esporozoítos de P. vivax e suas variantes, P. falciparum e P. malariae, que poderiam decorrer de contatos prévios, recentes ou não, com plasmódios humanos ou símios (o ciclo silvestre foi evidenciado em estudo paralelo realizado na mesma área). Por outro lado, estes resultados sugerem que, como em outros lugares onde existem vetores potenciais da malária, as condições epidemiológicas poderiam potencialmente permitir a transmissão da malária na área de influência do lago de Porto Primavera, se indivíduos infectados fossem introduzidos em número suficiente. Estudos adicionais deverão ser realizados para elucidar os fenômenos relatados neste artigo

Anthropophilic activity of Aedes aegypti and of Aedes albopictus in area under control and surveillance

OBJECTIVE: To describe the hematophagous activity of Ae. aegypti and Ae. albopictus in an area under control and surveillance. METHODS: The study was conducted during 18 months, from April, 1993 to October, 1994, in Cosmópolis, São Paulo state, Brazil. Human baits were used to collect mosquitoes. The number of females captured is presented monthly by area of the city and local in the household. The rainfall was measured and indices are presented without model adjustment. RESULTS: The presence of females of both species was observed in 83% (Ae albopictus) and 61% (Ae. aegypti) of the period studied. The months of January, February and March presented the highest rates of activity for females of both species, with Ae. albopictus being more frequently captured than Ae. aegypti. Both species were captured in central and peripheral areas of the city, during the day from 9 to 12 am and from 4 to 7 pm. By a Poisson regression, it was observed that Ae. albopictus females were more frequently captured in the peri-housed area, in an independent way considering the area of the city. CONCLUSIONS: It has been identified difference on the hematophagous activity only for Ae. albopictus, being of importance the outside area of the house.OBJETIVO: Descrever a atividade de hematofagia de Aedes aegypti e Aedes albopictus em área submetida ao controle e à vigilância entomológica. MÉTODOS: O estudo foi realizado de abril de 1993 a setembro de 1994, na cidade de Cosmópolis, Estado de São Paulo. Utilizou-se isca humana para coleta dos exemplares. Calculou-se o número médio mensal de fêmeas capturadas por domicílio segundo as áreas central e periférica do município e os locais intra e peridomiciliar. Apresenta-se o índice pluviométrico mensal sem ajuste de modelo. RESULTADOS: Observou-se a presença de fêmeas das espécies em 83% (Ae. albopictus) e 61% (Ae. aegypti) do período estudado. Os meses de janeiro, fevereiro e março apresentaram maior atividade de fêmeas para as duas espécies, com maior presença de Ae. albopictus do que de Ae. aegypti. Os exemplares foram capturados na área central e periférica da cidade, nos períodos matutino, das 9:00 às 12:00 e crepuscular vespertino, das 16:00 às 19:00. Com utilização de modelo de regressão de Poisson, observou-se que fêmeas de Ae. albopictus foram mais capturadas no peridomicílio, independente se área central ou periférica. CONCLUSÕES: Foi detectada diferença no perfil de atividade da hematofagia somente para o Ae. albopictus, sendo de relevância o peridomicílio

Mosquitoes infected with dengue viruses in Brazil

Dengue epidemics have been reported in Brazil since 1985. The scenery has worsened in the last decade because several serotypes are circulating and producing a hyper-endemic situation, with an increase of DHF/DSS cases as well as the number of fatalities. Herein, we report dengue virus surveillance in mosquitoes using a Flavivirus genus-specific RT-Hemi-Nested-PCR assay. The mosquitoes (Culicidae, n = 1700) collected in the Northeast, Southeast and South of Brazil, between 1999 and 2005, were grouped into 154 pools. Putative genomes of DENV-1, -2 and -3 were detected in 6 mosquito pools (3.8%). One amplicon of putative DENV-1 was detected in a pool of Haemagogus leucocelaenus suggesting that this virus could be involved in a sylvatic cycle. DENV-3 was found infecting 3 pools of larvae of Aedes albopictus and the nucleotide sequence of one of these viruses was identified as DENV-3 of genotype III, phylogenetically related to other DENV-3 isolated in Brazil. This is the first report of a nucleotide sequence of DENV-3 from larvae of Aedes albopictus

Contribuição ao estudo da história natural de enteroparasitoses em uma comunidade fechada 1: Prevalência de enteroparasitos em uma comunidade fechada

Empregando 5 métodos de diagnóstico coprológico, os autores estudam a prevalência de enteroparasitas em um orfanato do município de Londrina, Paraná, discutindo os resultados