Culex quinquefasciatus vitellogenesis: morphological and biochemical aspects

The vitellogenic process in Culex quinquefasciatus, which is triggered by a blood meal, involves the synthesis, distribution and storage of the nutrients necessary for embryo development. The fat body of an adult female Cx. quinquefasciatus revealed two cell types: large trophocytes and small, eosinophilic, "oenocyte-like" cells, which show no morphological changes throughout the gonotrophic cycle. Trophocytes, which only begin to synthesise vitellogenin (Vg) 12 h post-blood meal (PBM), undergo a series of morphological changes following engorgement. These changes include the expansion of the rough endoplasmic reticulum (RER) and Golgi complex, which are later destroyed by autophagosomes. At 84 h PBM, trophocytes return to their pre-engorgement morphology. The ovarian follicles of non-blood-fed Cx. quinquefasciatus contain a cluster of eight undifferentiated cells surrounded by follicular epithelium. After engorgement, the oocyte membrane facing the perioocytic space increases its absorptive surface by microvilli development; large amounts of Vg and lipids are stored between 24 and 48 h PBM. Along with yolk storage in the oocyte, follicular cells exhibit the development of RER cisternae and electron-dense granules begin to fill the perioocytic space, possibly giving rise to endochorion. Later in the gonotrophic cycle, electron-dense vesicles, which are possible exochorion precursors, fuse at the apical membrane of follicular cells. This fusion is followed by follicular cell degeneration.

Wing diagnostic characters for Culex quinquefasciatus and Culex nigripalpus (Diptera, Culicidae)

Wing diagnostic characters for Culex quinquefasciatus and Culex nigripalpus (Diptera, Culicidae). Culex quinquefasciatus and Culex nigripalpus are mosquitoes of public health interest, which can occur sympatrically in urban and semi-urban localities. Morphological identification of these species may be difficult when specimens are not perfectly preserved. In order to suggest an alternative taxonomical diagnosis, wings of these species were comparatively characterized using geometric morphometrics. Both species could be distinguished by wing shape with accuracy rates ranging from 85-100%. Present results indicate that one can identify these species relying only on wing characters when traditional taxonomical characters are not visible.

Density and survival rate of Culex quinquefasciatus at Parque Ecológico do Tietê, São Paulo, Brazil

Parity rate, gonotrophic cycle length, and density of a Culex quinquefasciatus female population was estimated at the Parque Ecológico do Tietê (PET), São Paulo, Brazil. Adult Cx. quinquefasciatus females were collected from vegetation along the edges of a polluted drainage canal with the use of a battery-powered backpack aspirator from September to November 2005 and from February to April 2006. We examined 255 Cx. quinquefasciatus ovaries to establish the parity rate of 0.22 and determined the gonotrophic cycle length under laboratory conditions to be 3 and 4 days. From these data, we calculated the Cx. quinquefasciatus survival rate to be 0.60 and 0.68 per day. Density of the Cx. quinquefasciatus female (5.71 females per m2) was estimated based on a population size of 28,810 individuals divided by the sampled area of 5,040 m2. Results of all experiments indicate medium survivorship and high density of the Cx. quinquefasciatus female population. This species is epidemiologically relevant in the PET area and should be a target of the vector control program of São Paulo municipality

Genetic-morphometric variation in Culex quinquefasciatus from Brazil and La Plata, Argentina

Variation among natural populations of Culex (Culex) quinquefasciatus Say is associated with different vectorial capacities. The species Cx. quinquefasciatus is present in the equatorial, tropical and subtropical zones in the Brazilian territory, with intermediate forms between Cx. quinquefasciatus and Culex pipiens occurring in regions of latitudes around 33°-35°S. Herein, we studied geographically distinct populations of Cx. quinquefasciatus by genetic characterization and analysis of intra-specific wing morphometrics. After morphological analysis, molecular characterization of Cx. quinquefasciatus and intermediate forms was performed by polymerase chain reaction of the polymorphic nuclear region of the second intron of the acetylcholinesterase locus. Additionally, the morphology of adult female wings collected from six locations was analyzed. Wing centroid sizes were significantly different between some geographical pairs. Mean values of R2/R2+3 differed significantly after pairwise comparisons. The overall wing shape represented by morphometric characters could be divided into two main groupings. Our data suggest that Brazilian samples are morphologically and genetically distinct from the Argentinean samples and also indicated a morphological distinction between northern and southern populations of Brazilian Cx. quinquefasciatus. We suggest that wing morphology may be used for preliminary assessment of population structure of Cx. quinquefasciatusin Brazil

Atividade predatória de Helobdella triserialis lineata (Hirudinea: Glossiphonidae) sobre formas imaturas de Aedes fluviatilis e Culex quinquefasciatus (Diptera: Culicidae) em laboratório

Foi avaliada a atividade predatória, em laboratório, de Helobdella triserialis lineata (Hirudinea: Glossiphonidae) sobre ovos, larvas e pupas de Aedes fluviatilis e Culex quinquefasciatus (Diptera: Culicidae), bem como a influência da presença desses hirudíneos sobre o comportamento de oviposição das fêmeas das duas espécies de mosquitos. Experimentos adicionais foram feitos testando a influência da profundidade da água e da sua salinidade sobre a capacidade predatória dos hirudíneos. Nas condições do experimento, foi observada predação de larvas e pupas, porém não de ovos das duas espécies de dípteros. Número estatisticamente menor de desovas foi depositado por fêmeas de Cx. quinquefasciatus em recipientes que continham hirudíneos, não ocorrendo o mesmo com fêmeas de Ae. fluviatilis. As diferentes profundidades de água testadas não interferiram na atividade predatória de H. t. lineata e somente em concentrações acima de 3% de NaCl essa atividade mostrou-se bastante diminuída.

Avaliação da sensibilidade de adultos de Culex quinquefasciatus Say a inseticidas químicos de contato

A sensibilidade de adultos do pernilongo doméstico Culex quinquefasciatus a 5 inseticidas químicos foi avaliada sob condições de laboratório pelo critério de Tempo Letal Mediano (TL50). Foram utilizados o organofosforado Malathion e quatro piretróides: Bifenthrin, Deltamethrin, Esfenvalerate e Alfamethrin. Foi sugerida uma técnica simples e eficiente para se avaliar adultos de um dia de idade incluindo 5 repetições para cada tratamento. Os resultados obtidos mostraram ser o método bastante adequado para avaliações rotineiras. Não ocorreu resistência a esses 5 princípios ativos, na população natural de Culex quinquefasciatus estudada.

Resistência a inseticidas organofosforados e carbamatos em população de Culex quinquefasciatus

Relata-se a ocorrência, na população de Culex quinquefasciatus, de resistência a pelo menos dois organofosforados e a um carbamato (malathion, fenitrothion e propoxur, respectivamente), mostrando a necessidade de haver programa de manejo de inseticidas. A suscetibilidade dessa população ao piretróide permetrina indica essa classe como de escolha numa eventual necessidade de substituição dos organofosforados.

Susceptibilidade larval de populações de Aedes aegypti e Culex quinquefasciatus a inseticidas químicos

OBJETIVO: Avaliar a susceptibilidade a inseticidas químicos de larvas de Culex quinquefasciatus e Aedes aegypti, provenientes de áreas sujeitas ou não a tratamentos de controle. MÉTODOS: Foram coletadas larvas de mosquitos em uma área não sujeita a tratamentos com inseticidas (Campinas, SP) e em áreas sujeitas a esses tratamentos (Campo Grande, MS e Cuiabá, MT). Foram usados bioensaios com concentrações diagnóstico e concentrações múltipla de inseticidas organofosforados e piretróides, segundo padrão da Organização Mundial de Saúde, para avaliar a susceptibilidade dessas larvas. RESULTADOS: Ensaios com larvas de Culex quinquefasciatus de Campinas, SP, permitiram a suspeita de resistência à cipermetrina e evidenciaram resistência à ciflutrina. Larvas dessa espécie coletadas em Campo Grande, MS, e Campinas, SP, apresentaram resistência ao temephos. Para a colônia campineira desta espécie, foram estabelecidas as razões de resistência: RR50=6,36 e RR95=4,94, com base em linhagem susceptível padrão. Adicionalmente, os testes com Aedes aegypti mostraram susceptibilidade similar ao temephos em uma população de campo (Cuiabá, MT) e uma de laboratório. CONCLUSÕES: Os resultados indicam resistência a organofosforado e piretróides em Culex quinquefasciatus, evidenciando a necessidade de avaliações e monitoramento da efetividade dos inseticidas a serem usados nos programas de controle de mosquitos.

Bionomics of Culex quinquefasciatus within urban areas of Rio de Janeiro, Southeastern Brazil

OBJECTIVE: To evaluate density, parity rates, daily survival and longevity of natural populations of Culex quinquefasciatus in three neighborhoods with distinct socio-economic and infrastructure profiles. METHODS: Mosquito collections of the Culex quinquefasciatus species were performed weekly during two four month periods, from August to November 2008 (spring) and March to June 2009 (fall), in a favela (slum), a suburban area and a middle class area of Rio de Janeiro, Southeastern Brazil. Collections were performed with backpack aspirators, in 20 randomly selected houses in each area per week, during 15-20 minutes per house. Ovaries were removed from captured females and classified as initial, intermediary or final stage. Furthermore, females were dissected for determination of parity based on the condition of the tracheal system. Mosquito survival rate and longevity were estimated on a per month basis for each neighborhood. RESULTS: We collected a total of 2,062 Culex quinquefasciatus, but monthly vector density was not correlated with temperature and rainfall. We dissected the ovaries of 625 Culex quinquefasciatus, and overall, there was a higher proportion of nulliparous females during the dryer months, while gravid females were more frequent in rainy months. In the middle class neighborhood, the parity rate reached up to 93.75% with survivorship of 0.979. Lower parity and survival rates were obtained in the suburban area (as low as 36.4% parity and 0.711 daily survival). Up to 84.7% of Culex quinquefasciatus females could survive the eight day period needed to complete West Nile Virus incubation. CONCLUSIONS: The survival rate of Culex quinquefasciatus varied significantly between the neighborhoods. This suggests that vectorial capacity and disease transmission risk may vary greatly between different urban areas, which is potentially useful information for vector control programs.

Residential characteristics aggravating infestation by Culex quinquefasciatus in a region of Northeastern Brazil

OBJECTIVE: Analyse how basic sanitation conditions, water supply and housing conditions affect the concentration of Culex quinquefasciatus METHODS: Populations of C. quinquefasciatus in 61 houses in the municipality of Olinda, PE, were monitored between October 2009 and October 2010. Observations were carried out in homes without the presence of preferred breeding sites in order to identify characteristics that may be aggravating factors for the development of the mosquito. Five aggravating factors were analysed: vegetation cover surrounding the home, number of residents/home, water storage, sewage drainage and water drainage. These characteristics were analysed in terms of presence or absence and as indicators of the degree of infestation, which was estimated through monitoring the concentration of eggs (oviposition traps - BR-OVT) and adults (CDC light traps). RESULTS: Sewage drainage to a rudimentary septic tank or to the open air was the most frequent aggravating factor in the homes (91.8%), although the presence of vegetation was the only characteristic that significantly influenced the increase in the number of egg rafts (p = 0.02). The BR-OVT achieved positive results in 95.1% of the evaluations, with the presence of at least one egg raft per month. A total of 2,366 adults were caught, with a mosquito/room/night ratio of 32.9. No significant difference was found in the number of mosquitoes caught in the homes. CONCLUSIONS: Although the sanitation and water supply influence the population density of C. quinquefasciatus, residence features that are not usually considered in control measures can be aggravating factors in sustaining the mosquito population.

Effects of ivermectin on Culex quinquefasciatus larvae

The effects of ivermectin, a semi-synthetic drug widely used for treatment of livestock parasitic diseases, were observed on Culex quinquefasciatus larvae. Toxic effects and mortality evaluation were carried out after 5, 15, 30 and 60 minutes of exposure to 1, 5 or 10 ppm of ivermectin solutions. Observations were made 24 and 48 hours after the beginning of the experiment, and loss of mobility, progressive paralysis and high mortality of larvae were recorded. The observed effects of ivermectin on the mosquito larvae is probably correlated with chloride channel activation on cell membranes.

Low genetic diversity in Wolbachia-Infected Culex quinquefasciatus (Diptera: Culicidae) from Brazil and Argentina

Culex quinquefasciatus is a vector of human pathogens, including filarial nematodes and several viruses. Although its epidemiological relevance is known to vary across geographical regions, an understanding of its population genetic structure is still incipient. In light of this, we evaluated the genetic diversity of Cx. quinquefasciatus and Cx. pipiens x Cx. quinquefasciatus hybrids collected from nine localities in Brazil and one site in Argentina. We used mitochondrial genes cox1 and nd4, along with the coxA and wsp genes of the maternally-inherited Wolbachia endosymbiont. The nd4 fragment was invariant between samples, whilst cox1 exhibited four haplotypes that separated two types of Cx. quinquefasciatus, one clustered in southern Brazil. Low sequence diversity was generally observed, being discussed. Both Brazilian and Argentinian mosquitoes were infected with a single Wolbachia strain. As reported in previous studies with these populations, cox1 and nd4 diversity is not congruent with the population structure revealed by nuclear markers or alar morphology. Future Cx. quinquefasciatus research should, if possible, evaluate mtDNA diversity in light of other markers.

Suscetibilidade de larvas de Culex quinquefasciatus a diferentes inseticidas

INTRODUÇÃO: O presente estudo teve como objetivo avaliar a suscetibilidade de larvas de Culex quinquefasciatus a dois piretróides (Cipermetrina e Deltametrina), dois derivados da Avermectina (ivermectina e abamectina) e a um organofosforado (Temefós). MÉTODOS: Larvas de 3º e 4º instares de C. quinquefasciatus foram expostas a diferentes concentrações destes (onze repetições) seguindo o protocolo da Organização Mundial de Saúde. Uma hora após a exposição, as larvas foram lavadas em água desclorada, transferidas para recipientes plásticos contendo água sem cloro, alimentadas e observadas por períodos de 24h, até se transformarem em adultos. Para a determinação das concentrações letais, os valores foram submetidos à análise de regressão usando o modelo probit pelo programa Minitab 15. RESULTADOS: Diferenças entre as estimativas da CL50 e CL90 justificaram que a população de mosquitos testada apresenta heterogeneidade em resposta aos inseticidas, sendo a maior concentração utilizada para a CL50, a partir da análise de probit para o Temefós. Todos os inseticidas avaliados causaram mortalidade mais acentuada nas primeiras 24h exceto quando expostas à ivermectina. CONCLUSÕES: As larvas são suscetíveis a todos os inseticidas testados e há uma necessidade de um monitoramento dos inseticidas utilizados.