Experimental Infection of Aedes albopictus (Diptera: Culicidae) Larvae with the Xiphidiocercariae of a Hematolechid

Aedes albopictus larvae were exposed, either individually or in groups, to different concentrations of xiphidiocercariae of Haematoloechus sp. for parasitological studies. It was observed the acute lethal effect and some aspects of the host-parasite relationship, such as delay or progress in the host life cycle, the number and location of the metacercariae in the host, adult host malformations and the amount of metacercariae required to cause death. A delay in the cycle and a high mortality rate was in general observed. Inside the larvae, the metacercariae were found predominantly in the thorax, abdominal segments and in the head, along with a reduced number in the anal lobe and cervix. It was shown that in addition to the quantity of metacercariae present, their location in the larvae was also relevant in the determination of mortality and anomalies. Malformed adults developed from larvae containing from one to three metacercariae.

Genetic diversity in Brazilian populations of Aedes albopictus

Random amplified polymorphic DNA (RAPD) analysis technique was undertaken in Aedes albopictus populations from three states in Brazil, Rio de Janeiro (RJ), Minas Gerais (MG) and Pernambuco (PE), to estimate the level of genetic variability and levels of genetic exchange between populations. Allele and genotype frequencies were measured on 47 RAPD loci. Average observed heterozigosity (Ho) ranged from 0.282 in MG to 0.355 in Casa Forte (PE) population. Genetic distances estimates indicated that RJ and MG were more genetically similar than populations from PE. Genetic variation observed in local Brazilian populations was attributed to genetic drift associated with restricted gene flow in recently established populations.

Comparative studies on the biology and filarial susceptibility of selected blood-feeding and autogenous Aedes togoi sub-colonies

Blood-feeding and autogenous sub-colonies were selected from a laboratory, stock colony of Aedes togoi, which was originally collected from Koh Nom Sao, Chanthaburi province, Southeast Thailand. Comparative biology and filarial susceptibility between the two sub-colonies (blood-feeding: F11, F13; autogeny: F38, F40) were investigated to evaluate their viability and vectorial capacity. The results of comparison on biology revealed intraspecific differences, i.e., the average egg deposition/gravid female (F11/F38; F13/F40), embryonation rate (F13/F40), hatchability rate (F11/F38; F13/F40), egg width (F11/F38), wing length of females (F13/F40), and wing length and width of males (F11/F38) in the blood-feeding sub-colony were significantly greater than that in the autogenous sub-colony; and egg length (F11/F38) and width (F13/F40), and mean longevity of adult females (F11/F38) and males (F13/F40) in the blood-feeding sub-colony were significantly less than that in the autogenous sub-colony. The results of comparison on filarial susceptibility demonstrated that both sub-colonies yielded similar susceptibilities to Brugia malayi [blood-feeding/autogeny = 56.7% (F11)/53.3%(F38), 60%(F13)/83.3%(F40)] and Dirofilaria immitis [blood-feeding/autogeny = 85.7%(F11)/75%(F38), 45%(F13)/29.4%(F40)], suggesting autogenous Ae. togoi sub-colony was an efficient laboratory vector in study of filariasis.

Effect of the Aedes fluviatilis saliva on the development of Plasmodium gallinaceum infection in Gallus (gallus) domesticus

Effect of Aedes fluviatilis saliva on the development of Plasmodium gallinaceum experimental infection in Gallus (gallus) domesticus was studied in distinct aspects. Chickens subcutaneously infected with sporozoites in the presence of the mosquito salivary gland homogenates (SGH) showed higher levels of parasitaemia when compared to those ones that received only the sporozoites. However, the parasitaemia levels were lower among chickens previously immunized by SGH or non-infected mosquito bites compared to the controls, which did not receive saliva. High levels of anti-saliva antibodies were observed in those immunized chickens. Moreover, 53 and 102 kDa saliva proteins were recognized by sera from immunized chickens. After the sporozoite challenge, the chickens also showed significant levels of anti-sporozoite antibodies. However, the ability to generate anti-sporozoites antibodies was not correlated to the saliva immunization. Our results suggest that mosquito saliva components enhance P. gallinaceum parasite development in naive chickens. However, the prior exposure of chickens to salivary components controls the parasitemia levels in infected individuals.

Efficiency of rubidium marking in Aedes albopictus (Diptera: Culicidae): preliminary evaluation on persistence of egg labeling, survival, and fecundity of marked female

Rubidium chloride (RbCl) has been used for the study of vector biology and behavior, although the efficacy of marking, egg production, and survivorship of marked females have been poorly studied. Four concentrations of RbCl were tested, among which 0.025 M was the best for marking Aedes albopictus: more than 80% of egg batches of females fed once with blood containing RbCl were marked; Rb-marked egg batches, interspersed with non marked ones were recovered until 61 days after a blood meal containing RbCl followed by non marked meals; RbCl was essentially detected in the abdomen of marked females, whose egg production and survivorship did not differ from non marked ones, at least in the three weeks following the Rb-marked blood meal.

Germline transformation of Aedes fluviatilis (Diptera:Culicidae) with the piggyBac transposable element

The technique to generate transgenic mosquitoes requires adaptation for each target species because of aspects related to species biology, sensitivity to manipulation and rearing conditions. Here we tested different parameters on the microinjection procedure in order to obtain a transgenic Neotropical mosquito species. By using a transposon-based strategy we were able to successfully transform Aedes fluviatilis (Lutz), which can be used as an avian malaria model. These results demonstrate the usefulness of the piggyBac transposable element as a transformation vector for Neotropical mosquito species and opens up new research frontiers for South American mosquito vectors.

Failure to demonstrate experimental vertical transmission of the epidemic strain of Chikungunya virus in Aedes albopictus from La Réunion Island, Indian Ocean

Aedes albopictus was responsible for transmission in the first outbreak of chikungunya (CHIK) on La Réunion Island, Indian Ocean, in 2005-2006. The magnitude of the outbreak on this island, which had been free of arboviral diseases for over 30 years, as well as the efficiency of Ae. albopictus as the main vector, raises questions about the maintenance of the CHIK virus (CHIKV) through vertical transmission mechanisms. Few specimens collected from the field as larvae were found to be infected. In this study, Ae. albopictus originating from La Réunion were orally infected with a blood-meal containing 10(8) pfu/mL of the CHIKV epidemic strain (CHIKV 06.21). Eggs from the first and second gonotrophic cycles were collected and raised to the adult stage. The infectious status of the progeny was checked (i) by immunofluorescence on head squashes of individual mosquitoes to detect the presence of viral particles or (ii) by quantitative RT-PCR on mosquito pools to detect viral RNA. We analysed a total of 1,675 specimens from the first gonotrophic cycle and 1,709 from the second gonotrophic cycle without detecting any viral particles or viral RNA. These laboratory results are compared to field records.

High prevalence and lack of diversity of Wolbachia pipientis in Aedes albopictus populations from Northeast Brazil

The use of Wolbachia as a tool to control insect vectors has recently been suggested. In this context, studies on the prevalence and diversity of this bacterium in wild populations are relevant. Here, we evaluated the diversity of two Wolbachiagenes (ftsZ and wsp) and the prevalence of this endosymbiont in wild Aedes albopictus. Using semi-nested polymerase chain reaction, our results showed that 99.3% of the individuals were superinfected with Wolbachia. In regards to genetic diversity, the two genes showed no variation within or among mosquito populations. An analysis of other Wolbachia markers may help to clarify the relationship between insect and endosymbiont.

Temporal variation of wing geometry in Aedes albopictus

Although native to the tropical and subtropical areas of Southeast Asia, Aedes albopictus is now found on five continents, primarily due to its great capacity to adapt to different environments. This species is considered a secondary vector of dengue virus in several countries. Wing geometric morphometrics is widely used to furnish morphological markers for the characterisation and identification of species of medical importance and for the assessment of population dynamics. In this work, we investigated the metric differentiation of the wings of Ae. albopictus samples collected over a four-year period (2007-2010) in São Paulo, Brazil. Wing size significantly decreased during this period for both sexes and the wing shape also changed over time, with the wing shapes of males showing greater differences after 2008 and those of females differing more after 2009. Given that the wings play sex-specific roles, these findings suggest that the males and females could be affected by differential evolutionary pressures. Consistent with this hypothesis, a sexually dimorphic pattern was detected and quantified: the females were larger than the males (with respect to the mean) and had a distinct wing shape, regardless of allometric effects. In conclusion, wing alterations, particularly those involving shape, are a sensitive indicator of microevolutionary processes in this species.

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.

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.

Effect of prostaglandin A1 in the induction of stress proteins in Aedes albopictus cells

Prostaglandins are natural fatty acid derivatives with diverse physiological effects, including immune function and the control of cell growth. While the action of prostaglandins in the induction of stress proteins in vertebrate cells is well documented, their functions in invertebrate cells have been poorly investigated. The purpose of the present study was to investigate the effect of prostaglandin A1 (PGA1; 0.25, 1.25 and 12.5 µg/ml) on protein synthesis during the growth of Aedes albopictus cells. We found that PGA1 stimulates the synthesis of several polypeptides with molecular masses of 87, 80, 70, 57, 29, 27 and 23 kDa in Aedes albopictus cells. When the proteins induced by PGA1 and those induced by heat treatment were compared by polyacrylamide gel electrophoresis, PGA1 was found to induce the stress proteins. The HSP70 family and the low-molecular weight polypeptides (29 and 27 kDa, respectively) were induced by PGA1 in the lag phase. We also observed that PGA1 is able to induce a 23-kDa polypeptide independently of the growth phase of the cell

Investigações sôbre o comportamento de formas adultas de mosquitos Silvestres no Estado de Sao Paulo, Brasil

Apresentaram-se os resultados das observações sobre o comportamento de formas adultas de Culicidae silvestres na área de Casa Grande, de zona de florestas primitivas situada na parte nordeste do Estado de São Paulo, Brasil. As investigações focalizaram os mosquitos de hábitos diurnos e crepusculares-noturnos, durante o período de 1963 a 1966. Foram obtidos dados concernentes à composição especifica, distribuição, ciclo anual da densidade, estratificação vertical na floresta e possíveis relações com o ambiente doméstico. A região estudada é considerada como bioma ou ecossistema do Anopheles (Kerteszia) cruzii e, dessa maneira, êsse mosquito compareceu de maneira predominante nas observações, merecendo por isso especial atenção. As outras espécies distribuiram-se predominantemente entre os Sabethini, além de alguns Culicini. No que concerne à domesticidade, o Anopheles cruzii apresentou-se como apreciável freqüentador das casas, seguido pelo Aedes serratus. Além disso, aquele anofelino mostrou-se acentuadamente acrodendrófilo, parecendo ser dotado de alguma ornitofilia.

Estudos ecológicos sobre mosquitos Culicidae no sistema da Serra do Mar, Brasil: 1 - Observações no ambiente extradomiciliar

Apresentam-se os resultados das coletas extradomiciliares de mosquitos Culicidae no Vale do Ribeira, Estado de São Paulo, Brasil. As observações foram levadas a efeito visando obter informações que esclareçam o mecanismo de transmissão de encefalite por vírus ocorrida em caráter epidêmico na região. Com o emprego de várias técnicas, foram feitas coletas em áreas com aspectos variáveis em relação à conservação do ambiente. Fundamentalmente, a diferenciação baseou-se em áreas alteradas, com matas residuais e ocorrência de casos de encefalite, e áreas conservadas, com matas primárias e sem a presença da doença, até o momento. Foram obtidos dados que sugerem possível associação entre a ocorrência da virose e o comportamento de Aedes serratus, Aedes scapularis e Culex (Melanoconion) sp. Admite-se também a possível participação de representantes de Limatus e Psorophora, embora provavelmente de maneira variável e menos intensa do que a dos anteriores. A possível ocorrência de ciclos prolongados de baixa densidade daqueles mosquitos na zona florestal da escarpa serrana, poderá explicar o possível papel de barreira que a Serra de Paranapiacaba e do Mar desempenham para a propagação da epidemia em direção ao planalto do Estado de São Paulo.