DsRed2 transient expression in Culex quinquefasciatus mosquitoes

Culex quinquefasciatus mosquitoes have been successfully genetically modified only once, despite the efforts of several laboratories to transform and establish a stable strain. We have developed a transient gene expression method, in Culex, that delivers plasmid DNA directly to the mosquito haemolymph and additional tissues. We were able to express DsRed2 fluorescent protein in adult Cx. quinquefasciatus mosquitoes by injecting plasmids directly into their thorax. The expression of DsRed2 in adult Cx. quinquefasciatus mosquitoes is an important stepping stone to genetic transformation and the potential use of new control strategies and genetic interactions.

COI barcode versus morphological identification of Culex ( Culex ) (Diptera: Culicidae) species: a case study using samples from Argentina and Brazil

Sequences of the cytochrome c oxidase subunit I (COI) mitochondrial gene from adults of 22 Culex ( Culex ) species from Argentina and Brazil were employed to assess species identification and to test the usefulness of COI for barcoding using the best close match (BCM) algorithm. A pairwise Kimura two-parameter distance matrix including the mean intra and interspecific distances for 71 COI barcode sequences was constructed. Of the 12 COI lineages recovered in the Neighbour-joining topology, five confirmed recognised morphological species ( Cx. acharistus , Cx. chidesteri , Cx. dolosus , Cx. lygrus and Cx. saltanensis ) with intraspecific divergences lower than 1.75%. Cx. bilineatus is formally resurrected from the synonymy of Cx. dolosus . Cx. maxi , Cx. surinamensis and the Coronator group species included were clustered into an unresolved lineage. The intraspecific distance of Cx. pipiens (3%) was almost twice the interspecific between it and Cx. quinquefasciatus (1.6%). Regarding the BCM criteria, the COI barcode successfully identified 69% of all species. The rest of the sequences, approximately 10%, 18% and 3%, remained as ambiguously, mis and unidentified, respectively. The COI barcode does not contain enough information to distinguish Culex ( Cux. ) species.

A comparative study of the effect of multiple immersions on Aedini (Diptera: Culicidae) mosquito eggs with emphasis on sylvan vectors of yellow fever virus

The effect of multiple immersions on Haemagogus janthinomys , Haemagogus leucocelaenus , Aedes albopictus and Ochlerotatus terrens eggs was studied. Eggs were collected in April, June, October and December of 2011 in Minas Gerais, Brazil. Most of the Aedes and Ochlerotatus eggs hatched upon the first immersion, while Haemagogus eggs showed a varied instalment hatching response. The number of immersions required for hatching increased for eggs collected closer to the dry winter season.

The remarkable journey of adaptation of the Plasmodium falciparum malaria parasite to New World anopheline mosquitoes

Plasmodium falciparum originated in Africa, dispersed around the world as a result of human migration and had to adapt to several different indigenous anopheline mosquitoes. Anophelines from the New World are evolutionary distant form African ones and this probably resulted in a more stringent selection of Plasmodium as it adapted to these vectors. It is thought that Plasmodium has been genetically selected by some anopheline species through unknown mechanisms. The mosquito immune system can greatly limit infection and P. falciparum evolved a strategy to evade these responses, at least in part mediated by Pfs47, a highly polymorphic gene. We propose that adaptation of P. falciparum to new vectors may require evasion of their immune system. Parasites with a Pfs47 haplotype compatible with the indigenous mosquito vector would be able to survive and be transmitted. The mosquito antiplasmodial response could be an important determinant of P. falciparum population structure and could affect malaria transmission in the Americas.

Updated distribution of Aedes albopictus (Diptera: Culicidae) in Spain: new findings in the mainland Spanish Levante, 2013

In 2004, Aedes (Stegomyia) albopictus (Skuse, 1894) was observed for the first time in Catalonia, northeastern Spain. A decade later, it has spread throughout the eastern Mediterranean region of the country and the Balearic Islands. Framed within a national surveillance project, we present the results of monitoring in 2013 in the autonomous communities of the mainland Levante. The current study reveals a remarkable increase in the spread of the invasive mosquito in relation to results from 2012; the species was present and well-established in 48 municipalities, most of which were along the Mediterranean coastline from the Valencian Community to the Region of Murcia.

The behaviour of mosquitoes in relation to humans under holed bednets: the evidence from experimental huts

The physical integrity of bednets is a concern of national malaria control programs, as it is a key factor in determining the rate of replacement of bednets. It is largely assumed that increased numbers of holes will result in a loss of protection of sleepers from potentially infective bites. Experimental hut studies are valuable in understanding mosquito behaviour indoors, particularly as it relates to blood feeding and mortality. This review summarises findings from experimental hut studies, focusing on two issues: (i) the effect of different numbers or sizes of holes in bednets and (ii) feeding behaviour and mortality with holed nets as compared with unholed nets. As might be expected, increasing numbers and area of holes resulted in increased blood feeding by mosquitoes on sleepers. However, the presence of holes did not generally have a large effect on the mortality of mosquitoes. Successfully entering a holed mosquito net does not necessarily mean that mosquitoes spend less time in contact with the net, which could explain the lack in differences in mortality. Further behavioural studies are necessary to understand mosquito behaviour around nets and the importance of holed nets on malaria transmission.

Temporal abundance of Aedes aegypti in Manaus, Brazil, measured by two trap types for adult mosquitoes

A longitudinal study was conducted in Manaus, Brazil, to monitor changes of adult Aedes aegypti (L.) abundance. The objectives were to compare mosquito collections of two trap types, to characterise temporal changes of the mosquito population, to investigate the influence of meteorological variables on mosquito collections and to analyse the association between mosquito collections and dengue incidence. Mosquito monitoring was performed fortnightly using MosquiTRAPs (MQT) and BG-Sentinel (BGS) traps between December 2008-June 2010. The two traps revealed opposing temporal infestation patterns, with highest mosquito collections of MQTs during the dry season and highest collections of BGS during the rainy seasons. Several meteorological variables were significant predictors of mosquito collections in the BGS. The best predictor was the relative humidity, lagged two weeks (in a positive relationship). For MQT, only the number of rainy days in the previous week was significant (in a negative relationship). The correlation between monthly dengue incidence and mosquito abundance in BGS and MQT was moderately positive and negative, respectively. Catches of BGS traps reflected better the dynamic of dengue incidence. The findings help to understand the effects of meteorological variables on mosquito infestation indices of two different traps for adult dengue vectors in Manaus.

Mayaro virus and dengue virus 1 and 4 natural infection in culicids from Cuiabá, state of Mato Grosso, Brazil

This study aimed to verify the diversity of Culicidae species and their frequency of infection with flaviviruses and alphaviruses in Cuiabá, state of Mato Grosso, Brazil. Mosquitoes were captured with Nasci aspirators and hand net in 200 census tracts, identified alive at species level and pooled in one-20 (11,090 mosquitoes, 14 species). Female pools (n = 610) were subjected to multiplex seminested-reverse transcription-polymerase chain reaction (RT-PCR) for 11 flavivirus and five alphavirus. Positive pools were tested by single RT-PCR followed by nucleotide sequencing, by RT-PCR for E1 gene [Mayaro virus (MAYV)] and by inoculation in Vero cells (MAYV) or C6/36 cells (flaviviruses). One/171 Aedes aegypti was positive for dengue virus (DENV)-1, 12/403 Culex quinquefasciatus, and four/171Ae. aegypti for MAYV, which was isolated from two pools containing two nonengorged females of Ae. aegypti and two ofCx. quinquefasciatus. DENV-4 was detected in 58/171 pools of Ae. aegytpi, 105/403 Cx. quinquefasciatus, two/five Psorophora sp., two/11 Psorophora varipes/Psorophora albigenu, one/one Sabethes chloropterus, two/five Culex bidens/Culex interfor, and one/one Aedes sp. DENV-4 was isolated from two pools containing three and 16 nonengorged Cx. quinquefasciatus females. Phylogenetic analysis revealed MAYV belongs to genotype L, clustering with human samples of the virus previously identified in the city. Cuiabá has biodiversity and ecosystem favourable for vector proliferation, representing a risk for arbovirus outbreaks.

Detection of all four dengue serotypes in Aedes aegypti female mosquitoes collected in a rural area in Colombia

The Aedes aegypti vector for dengue virus (DENV) has been reported in urban and periurban areas. The information about DENV circulation in mosquitoes in Colombian rural areas is limited, so we aimed to evaluate the presence of DENV in Ae. aegypti females caught in rural locations of two Colombian municipalities, Anapoima and La Mesa. Mosquitoes from 497 rural households in 44 different rural settlements were collected. Pools of about 20 Ae. aegypti females were processed for DENV serotype detection. DENV in mosquitoes was detected in 74% of the analysed settlements with a pool positivity rate of 62%. The estimated individual mosquito infection rate was 4.12% and the minimum infection rate was 33.3/1,000 mosquitoes. All four serotypes were detected; the most frequent being DENV-2 (50%) and DENV-1 (35%). Two-three serotypes were detected simultaneously in separate pools. This is the first report on the co-occurrence of natural DENV infection of mosquitoes in Colombian rural areas. The findings are important for understanding dengue transmission and planning control strategies. A potential latent virus reservoir in rural areas could spill over to urban areas during population movements. Detecting DENV in wild-caught adult mosquitoes should be included in the development of dengue epidemic forecasting models.

Influência da temperatura sobre a longevidade, fecundidade e atividade hematofágica de Aedes (Stegomyia) albopictus Skuse, 1894 (Diptera, Culicidae) sob condições de laboratório

Influence of temperature on longevity, fecundity, and haematophagic activity of Aedes (Stegomyia) albopictus Skuse, 1894 (Diptera, Culicidae) under laboratory condition. Based on adults of Aedes albopictus maintained under four constant temperatures, 15ºC, 20ºC, 25ºC and 30ºC, longevity of males and females, fecundity, eggs viability, number of days from emergence to the first bloodsucking, and duration of the preoviposition period were observed. The average of the longevity for female was 19.45, 59.35, 61.55 and 42.72 days, under 15ºC, 20ºC, 25ºC and 30ºC, respectively. The greatest average number of bloodsucking per female was 8.32, under 25ºC. The average number of eggs per female was 1.00, 73.91, 269.87 and 183.69, and the greatest rates of viable eggs per female was 25ºC and 30ºC. The haematophagic activities and the oviposition were significantly influenced by temperature.

Exigências térmicas de Aedes (Stegomyia) albopictus Skuse, 1894 (Diptera, Culicidae) em condições de laboratório

Thermal requirements of Aedes (Stegomyia) albopictus Skuse, 1894 (Diptera, Culicidae) under laboratory conditions. Allochtonous species of Ae. albopictus in the American continent can be found in fourteen Brazilian states - about fifteen years had been elapsed since the first report of the presence of this species in Brazilian territory. Considering its potential epidemiological importance and its adaptation to several habitats, it was determined, for this species, the threshold temperature and the thermal constant for egg, larval and pupal stages under laboratory conditions under four constant temperatures and 12:12 hours light-dark photoperiod. The threshold temperature for the egg phase and for the first instar were quite similar: 9.07 ºC (K=214.46 degree days) and 9.23 ºC (K= 36.64 degree days), respectively. For 2nd, 3rd and 4th instar, the basal-temperature was higher, oscilating between 12.26 ºC and 13.95 ºC. The threshold temperature for the complete larval stage and for the pupal stage were 12.03 ºC (K= 99.48 degree days) and 11.87 ºC (K=32.40 degree days) for males and 11.95 ºC (K=110.11 degree days) and 11.60 ºC (K=35.30 degree days) for females, respectively.

Ultrastructure of the eggs of two species of Anopheles (Anopheles) Meigen (Diptera, Culicidae)

Egg ultrastructure of two morphologicaly similar species of the Arribalzagia Series, Anopheles (A.) costai Fonseca & Ramos, 1939 and A. (A.) mediopunctatus (Lutz, 1903) are described and illustrated using scanning electron micrographs. Although similar in the adult stage, male genitalia structures and larval and pupal stages, the eggs of these species are distinct. The eggs of A. costai and A. mediopunctatus are compared with that of A. forattinii Wilkerson & Sallum, 1999. Incomplete illustration of egg of A. forattinii is provided for comparison.

O acervo de mosquitos (Diptera, Culicidae) de Nelson L. Cerqueira na Coleção de Invertebrados do Instituto Nacional de Pesquisas da Amazônia, Manaus, Brasil

Registra-se a descoberta de espécimes de mosquitos que pertenciam a Nelson L. Cerqueira, e estão sendo depositados na coleção de Invertebrados do Instituto Nacional de Pesquisas da Amazônia. A coleção contém 2.046 espécimes adultos e 387 lâminas representando 261 espécies, 22 gêneros, incluindo 51 parátipos de 34 espécies. Mais de 90% dos espécimes foram coletados no Brasil dos quais metade são do Estado do Amazonas. As espécies representadas neste acervo são listadas indicando o número de espécimes para cada tipo de preparação e as localidades de coleta. O material tipo também é listado, incluindo os dados dos rótulos de identificação e de procedência, bem como outras informações pertinentes.

Checklist of aedine mosquito species (Diptera, Culicidae, Aedini) occurring in Middle and South America (south of the United States) reflecting current generic and subgeneric status

A list of species of tribe Aedini from Middle and South American, south of the United States, with their current generic combinations is provided. Its purpose is to enable entomologists, public health personnel and mosquito control workers to more quickly become familiar with recent formal taxonomic changes within the tribe.

Culicinae (Diptera, Culicidae) ocorrentes no Estado do Rio Grande do Sul, Brasil

Este trabalho lista as espécies e aponta a ocorrência de Culicinae, com base em coletas de adultos realizadas no período de setembro de 2001 a agosto de 2002, em 11 zonas fisiográficas do Estado, busca de informações bibliográficas e verificação de exemplares em coleções. São relacionadas 57 espécies, sendo que 22 constituem novas ocorrências para o Rio Grande do Sul. São ampliados os locais de ocorrência de Aedeomyia squamipennis, Culex chidesteri, Culex quinquefasciatus, Haemagogus leucocelaenus, Mansonia titillans, Ochlerotatus albifasciatus, Ochlerotatus crinifer, Ochlerotatus fluviatilis, Ochlerotatus scapularis, Ochlerotatus serratus, Psorophora confinnis, Psorophora ferox, Sabethes aurescens, Trichoprosopon pallidiventer e Uranotaenia lowii.