Description of immature stages of Culex ocossa Dyar & Knab, Culex delpontei Duret and Culex pereyrai Duret of the Melanoconion subgenus (Diptera: Culicidae)

The larva and pupa of Culex (Melanoconion) ocossa Dyar & Knab are redescribed and those of Culex (Melanoconion) delpontei Duret and Culex (Melanoconion) pereyrai Duret are described from specimens collected in the states of São Paulo and Paraná, Brazil. The pupa of Cx. ocossa differs from those of the other two species in having seta 5-IV-VI dark with strongly aciculated branches, and caudolateral angle of segment VIII produced into sharp point, and seta 1-P present; Cx. delpontei can be distinguished from Cx. pereyrai in possessing paddle lightly tanned, trumpet flared, and wing and leg cases lightly tanned, without pattern of dark spots; Cx. pereyrai can be recognized by having wing case with pattern of dark, discontinuously pigmented, longitudinal lines, and trumpet cylindrical, not flared. The larvae of the three species share the presence of seta 2-C placed medially to seta 1-C.

Observações sobre domiciliação de mosquitos Culex (Melanoconion), em ambiente com acentuadas modificações antrópicas

Estudam-se alguns aspectos do comportamento de espécies de Culex (Melanoconion) em ambiente antrópico intensamente modificado. Foram realizadas coletas mensais de adultos, concomitantes e levadas a efeito no peridomicílio, em área aberta e em área coberta por vegetação de segunda formação. Foram focalizadas as espécies dominantes e representadas por Cx. delpontei, Cx. ocossa, Cx. ribeirensis, Cx. sacchettae e Cx. taeniopus. No peridomicílio, destacam-se pela regularidade de sua presença, Cx. ribeirensis e Cx. sacchettae, incluindo apreciável comparecimento às coletas com isca humana ali realizadas. A época de maior rendimento das capturas correspondeu ao primeiro trimestre, em especial modo, ao mês de março. Aliado a esse aspecto, Cx. ribeirensis e Cx. sacchettae revelaram apreciável freqüência à isca humana. Em relação a esta, o menor comparecimento foi apresentado por Cx. delpontei e Cx. ocossa. Quanto a Cx. taeniopus, compareceu de maneira um tanto irregular e em número geralmente inferior ao dos demais. As observações evidenciaram curva de atividade aumentando rapidamente por ocasião do início da noite e mantendo-se durante todo o período noturno, apresentada por Cx. ribeirensis, Cx. sacchettae e Cx. taeniopus. O outro tipo de curva foi observado com Cx. delpontei e Cx. ocossa, com aumento gradual na primeira metade da noite, alcançando o máximo ao redor da meia-noite, e declinando sensivelmente na segunda metade do período. As coletas de formas imaturas permitiram evidenciar criadouros de Cx. delpontei em cursos de água de porte médio ou grande, e associados à vegetação aquática flutuante. São feitas considerações de ordem ecológica passíveis de interpretar esses comportamentos de interesse epidemiológico.

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.

Culex nigripalpus Theobald (Diptera, Culicidae) feeding habit at the Parque Ecológico do Tietê, São Paulo, Brazil

The blood feeding of a population of Cx. nigripalpus from Parque Ecológico do Tietê (PET) was investigated using an indirect ELISA protocol. Mosquitoes were captured outside houses. Five hundred sixteen engorged females collected in a reforested area and 25 in an open area were tested. Rodents and dogs were the most common blood sources, accounting for approximately 65.3% of blood meals. Human blood was detected in 10.9%, dog blood in 26.1%, chicken blood in 2.4%, and rodent blood in 39.2% of the 541 insects tested. ELISA failed in identifying the blood sources of 233 engorged females, indicating that the mosquitoes may have fed on a host which was not tested. One hundred six individuals were positive for more than one host. The unweighted human blood index was 0.14 and the rodent/human, human/chicken, and dog/rodent feeding index values were 2.70, 1.51, and 1.33, respectively. Furthermore, rodents are defensive hosts for this haematophagous insect which looks for another host to complete blood-feeding. Considering that rodents are potential reservoirs for Mucambo virus and Saint Louis encephalitis virus and that Cx. nigripalpus feed on the blood of those mammals, we hypothesize that mosquito population in PET could participate in the transmission cycle of those arboviruses. Additionally, this species might be involved in the transmission of Dirofilaria immitis to dogs at this area.

First Record of Culex (Culex) brethesi (Dyar) (Diptera: Culicidae) in Rio Grande do Sul State, Brazil

This is the first record of Culex (Culex) brethesi (Dyar) in Rio Grande do Sul state, Brazil. The species was identified from specimens collected in a sand bar vegetation with the aid of a Nasci's trap, during an expedition for surveillance of the West Nile Virus in July of 2006, in the city of Mostardas, Rio Grande do Sul, Brazil.

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

Redescription of Culex (Culex) dolosus (Lynch Arribálzaga)(Diptera: Culicidae), based on specimens from Pico do Itapeva, Serra da Mantiqueira, São Paulo, Brazil

Culex (Culex) dolosus (Lych Arribálzaga, 1891) is re-described and compared with Cx. eduardoi Casal & Garcia (1968) based on morphological characteristics. Both species are morphologically similar, and they have been largely misidentified throughout Brazil. Adult male and female, fourth instar larvae, and pupae of Cx. dolosus were examined, based on specimens from Pico do Itapeva, Pindamonhangaba Municipality, Serra da Mantiqueira, São Paulo State, southern Brazil. Male genitalia, larvae and pupae are illustrated. Geographical distribution is summarized from published records, and information on bionomics is based on literature and field data

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

Japanese encephalitis on Badu Island, Australia: the first isolation of Japanese encephalitis virus from Culex gelidus in the Australasian region and the role of mosquito host-feeding patterns in virus transmission cycles

During investigation of an outbreak of Japanese encephalitis (JE) in the Torres Strait, Australia, in 2000, mosquitoes were collected in Badu Island community and at a newly established communal piggery about 3 km from the community. A total of 94285 mosquitoes, comprising 91240 (96.8%) unengorged females, 1630 (1.7%) blood-engorged females and 1415 (1.5%) males, were processed for virus isolation. One isolate of JE virus was obtained from Culex gelidus, with a minimum infection rate of 12.4:1000. This is the first isolate of JE virus from Cx. gelidus in the Australasian region. No isolates were obtained from Cx. annulirostris, the primary implicated Australian JE vector. Analysis of mosquito host-feeding patterns, using gel diffusion, demonstrated that Cx. annulirostris and 5 other species fed predominately on mammals, Analysis of blood-fed mosquitoes collected within the community demonstrated that the proportion of Cx. annulirostris feeding on pigs in 2000 (2.3%) was significantly lower than that for the 1995-97 period (31.3%). The removal of the pigs from Badu Island community has limited the contact between potential amplifying hosts and mosquitoes, thus potentially reducing the risk of transmission of JE virus to the human population.

Development and evaluation of a species diagnostic polymerase chain reaction-restriction fragment-length polymorphism procedure for cryptic members of the Culex sitiens (Diptera : Culicidae) subgroup in Australia and the southwest Pacific

Members of the Culex sitiens subgroup are important vectors of arboviruses, including Japanese encephalitis virus, Murray Valley encephalitis virus and Ross River virus. Of the eight described species, Cx. annulirostris Skuse, Cx. sitiens Wiedemann, and Cx. palpalis Taylor appear to be the most abundant and widespread throughout northern Australia and Papua New Guinea (PNG). Recent investigations using allozymes have shown this subgroup to contain cryptic species that possess overlapping adult morphology. We report the development of a polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) procedure that reliably separates these three species. This procedure utilizes the sequence variation in the ribosomal DNA ITS1 and demonstrates species-specific PCR-RFLP profiles from both colony and field collected material. Assessment of the consistency of this procedure was undertaken on mosquitoes sampled from a wide geographic area including Australia, PNG, and the Solomon Islands. Overlapping adult morphology was observed for Cx. annulirostris and Cx. palpalis in both northern Queensland and PNG and for all three species at one site in northwest Queensland.

Observações sobre os mosquitos Culex da cidade de São Paulo, Brasil

São apresentados os resultados obtidos na coleta de mosquitos do gênero Culex na área urbana da cidade de São Paulo, Brasil. Foram empregadas armadilhas luminosas automáticas tipo "New Jersey 50". Os resultados revelaram a presença de outras populações representadas principalmente por Culex chidesteri, C. dolosus e C. bidens as quais, em conjunto, chegaram algumas vezes a sobrepujar a de Culex pipiens fatigans. O maior rendimento foi obtido em áreas com abastecimento de água mas sem rede de esgotos. As coletas intradomiciliares revelaram franca predominância de C. pipiens fatigans.

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.