Aedes (Stegomyia) albopictus (SKUSE), su asociación con Aedes (Stegomyia) aegypti (LINNAEUS) y otros culícidos en Allende, N.L., México [por] Juan de Dios Aguilar Gueta

Tesis (Maestría en Ciencias con Especialidad en Entomología Médica) U.A.N.L.

Field prevalence of Wolbachia in the mosquito vector Aedes albopictus

The endosymbiotic bacteria in the genus Wolbachia have been proposed as a potential candidate to deliver pathogen-blocking genes into natural populations of medically important insects. The successful application of Wolbachia in insect vector control depends on the ability of the agent to successfully invade and maintain itself at high frequency under field conditions. Here, we evaluated the prevalence of Wolbachia infections in a field population of the Wolbachia-superinfected mosquito Aedes albopictus. A field prevalence of 100% (n = 1,016) was found in a single population in eastern Thailand via polymerase chain reaction (PCR) testing of Wolbachia both from individual parent females and their corresponding F1 offspring. This is the first report of accurate Wolbachia prevalence in a field population of an insect disease vector. The prevalence of superinfection was estimated to be 99.41%. All single-infected individual mosquitoes (n = 6) were found to harbor group A Wolbachia. For this particular population, none was found to be single-infected with group B Wolbachia. Our results also show that PCR testing of field materials alone without checking F1 offspring overestimated the natural prevalence of single infection. Thus, the confirmation of infection status by means of F1 offspring was critical to the accurate estimates of Wolbachia prevalence under field conditions.

Produtividade de criadouro de Aedes albopictus em ambiente urbano

INTRODUÇÃO: O encontro de Aedes albopictus na cidade de Cananéia, região Sudeste do Estado de São Paulo, Brasil, ensejou a ocasião de realizar observações que visassem avaliar a produtividade de criadouro grande e permanente. Como objetivo, após selecionar o habitat a ser estudado, tentou-se avaliar-lhe a contribuição para a densidade local do mosquito. MATERIAL E MÉTODO: Em área predeterminada procedeu-se a levantamento de criadouros potenciais. Constatada a presença da espécie, foi selecionado um dos recipientes que preenchia os requisitos desejados. O acompanhamento foi feito de maneira ininterrupta, no período de novembro de 1996 a maio de 1997. As observações obdeceram a ritmo quinzenal retirando, cada vez, amostra da água correspondente a 0,14, ou seja, um sétimo do volume total de 70 litros. Procurou-se coletar, identificar e numerar, por sexo, as pupas existentes. Concomitantemente, procedeu-se à captura de formas adultas. Foi utilizada a isca humana das 15:00 às 18:00h, instalada a cerca de 6 metros do mencionado criadouro. Finda essa coleta, foi feita aspiração com 30 min. de duração em locais de abrigo representados pela abundante vegetação circunjacente. RESULTADOS: Nas coletas de formas imaturas do criadouro, o Ae. albopictus compareceu com 44,9%. Ao longo de 15 amostras regularmente realizadas obteve-se a média de 31,13 pupas pertencentes a essa espécie. O índice de emergência(E) foi de 2,1. A multiplicação desse valor por sete forneceu a média diária de 14,7 fêmeas. Nas coletas de adultos desse sexo, a média de Williams para a isca humana foi de 30,7, enquanto a densidade média horária da aspiração dos locais de abrigo foi de 9,2. O cálculo do acúmulo diário concluiu pela presença de 22,8 fêmeas, por dia, capazes de freqüentar a isca humana, nessa situação e condições. DISCUSSÃO: A contagem de pupas possibilitou estimar a produtividade de criadouro de Ae. albopictus, tipo grande (dez litros ou mais) e de caráter permanente. A água acumulada no recipiente estudado apresentou-se rica em matéria orgânica, predominantemente de natureza vegetal. Neste particular, não há como compará-lo a reservatórios destinados ao armazenamento de água para uso doméstico. A falta de manutenção desses reservatórios poderá contribuir para levar a situação, se não idêntica, pelo menos próxima da evidenciada no trabalho. Embora se trate de espécie até agora não incriminada como vetora, é de se admitir que as observações encontradas possam ser utilizadas no programa de erradicação de Ae. aegypti que está em curso no Brasil.

Factors influencing the development of Plasmodium gallinaceum in Aedes fluviatilis

Aedes fluviatilis is susceptible to infection by Plasmodium gallinaceum and is a convenient insect host for the malaria parasite in countries where Aedees aegypti cannot be maintained in laboratories. In South America, for instance, the rearing of A. aegypti the main vector of urban yellow fever, is not advaisable because of the potential health hazard it represents. Our results of the comparative studies carried out between the sporogonic cycle produced with two lines of P. gallinaceum parasites into A. fuviatilis were as follows. As proved for A. aegypti, mosquito infection rates were variable when A. fluviatilis blood-fed on chicks infected with and old syringe-passaged strain of P. gallinaceum. Oocysts developed in 41% of those mosquitos and the mean peak of oocyst production was 56 per stomach. Salivary gland infections developed in about 6% of the mosquitos. The course of sporogony was unrelated to the size of the inoculum administered to chicks or to the route by which the birds were infected. The development of infected salivary glands was unrelated to oocyst production. Sporogony of P. gallinaceum was more uniform when mosquitos blood-fed on chicks infected with a sporozoite-passaged strain. Oocysts developed in about 50% of those mosquitoes and the mean peak of oocyst production was 138 per stomach, with some individuals having as many as 600-800 oocysts. Infected salivary glands developed in a mean of 27% of the mosquitos but, in some batches, was a high as 50%. Patterns of salivary gland parasitism were similar to those of oocyst production. The course of sporogony of P. gallinaceum in A. fluviatilis is analized in relation to degree of parasitemia and gametocytemia in the vertebrate host.

Dengue virus detection by using reverse transcription-polymerase chain reaction in saliva and progeny of experimentally infected Aedes albopictus from Brazil

Oral susceptibility and vertical transmission of dengue virus type 2 (DENV-2) in an Aedes albopictus sample from Rio de Janeiro was estimated. The infection (36.7%) and transmission (83.3%) rates for Ae. albopictus were higher than those of an Ae. aegypti colony used as control, 32.8 and 60%, respectively. Fourth instar larvae and females descendants of 48.5 and 39.1% of experimentally infected Ae. albopictus showed to harbor the virus. The oral susceptibility and the high capacity to assure vertical transmission exhibited by Ae. albopictus from Brazil reinforce that this species may play a role in the maintenance of the virus in nature and be a threat for dengue control in the country.

Updating the geographical distribution and frequency of Aedes albopictus in Brazil with remarks regarding its range in the Americas

The geographical distribution of Aedes albopictus in Brazil was updated according to the data recorded across the country over the last eight years. Countrywide house indexes (HI) for Ae. albopictus in urban and suburban areas were described for the first time using a sample of Brazilian municipalities. This mosquito is currently present in at least 59% of the Brazilian municipalities and in 24 of the 27 federal units (i.e., 26 states and the Federal District). In 34 Brazilian municipalities, the HI values for Ae. albopictus were higher than those recorded for Ae. aegypti, reaching figures as high as HI = 7.72 in the Southeast Region. Remarks regarding the current range of this mosquito species in the Americas are also presented. Nineteen American countries are currently infested and few mainland American countries have not confirmed the occurrence of Ae. albopictus. The large distribution and high frequency of Ae. albopictus in the Americas may become a critical factor in the spread of arboviruses like chikungunya in the new world.

Dimorfismo sexual alar em Aedes scapularis (Diptera: Culicidae)

A detecção do sexo de mosquitos da família Culicidae é importante em estudos faunísticos e epidemiológicos, pois somente as fêmeas possuem competência vetora para patógenos. O dimorfismo sexual de genitália e de apêndices cefálicos é, em geral, facilmente visível em culicídeos. As asas também podem ser dimórficas e assim poderiam complementar o procedimento de sexagem. No entanto, tal distinção não é facilmente notável à observação direta. Visando descrever formalmente o dimorfismo sexual alar em Aedes scapularis, um culicídeo vetorialmente competente para arbovírus e filárias, asas de machos e fêmeas foram comparadas usando-se métodos de morfometria geométrica e análise estatística multivariada. Nestas análises, populações dos municípios São Paulo e Pariquera-Açu (Estado de São Paulo) foram amostradas. A forma das asas mostrou evidente dimorfismo sexual, o que permitiu um índice de acurácia de 100% em testes-cegos de reclassificação, independentemente da origem geográfica. Já o tamanho alar foi sexualmente dimórfico apenas na população de São Paulo. Aparentemente, a forma alar é evolutivamente mais estável que o tamanho, interpretação que está de acordo com a teoria de Dujardin (2008b), de que a forma alar de insetos seria composta por caracteres genéticos quantitativos e pouco influenciada por fatores não-genéticos, enquanto que o tamanho alar seria predominantemente determinado por plasticidade decorrente de influências ambientais.

Registro de Aedes albopictus em áreas epizoóticas de febre amarela das Regiões Sudeste e Sul do Brasil (Díptera:Culicidae)

Durante estudo biológico e ecológico sobre mosquitos levado a cabo em área com registro de epidemia de febre amarela silvestre e epizootia em macacos, foram encontrados adultos de Aedes Albopictus. A tendência da espécie para invadir ambiente extradomiciliar potencializa a chance de infecção natural, ao tempo em que evolui para formar um elo entre focos naturais de vírus e o ambiente urbano. Esta Nota Técnica representa um alerta aos gestores dos três poderes públicos sobre perspectivas de mudanças no perfil epidemiológico atual da febre amarela no Brasil

Yellow Fever Virus in Haemagogus leucocelaenus and Aedes serratus Mosquitoes, Southern Brazil, 2008

Yellow fever virus (YFV) was isolated from Haemagogus leucocelaenus mosquitoes during an epizootic in 2001 in the Rio Grande do Sul State in southern Brazil In October 2008 a yellow fever outbreak was reported there with nonhuman primate deaths and human cases This latter outbreak led to intensification of surveillance measures for early detection of YFV and support for vaccination programs We report entomologic surveillance in 2 municipalities that recorded nonhuman primate deaths Mosquitoes were collected at ground level identified and processed for virus isolation and molecular analyses Eight YFV strains were isolated (7 from pools of Hg leucocelaenus mosquitoes and another from Aedes serratus mosquitoes) 6 were sequenced and they grouped in the YFV South American genotype I The results confirmed the role of Hg leucocelaenus mosquitoes as the main YFV vector in southern Brazil and suggest that Ae serratus mosquitoes may have a potential role as a secondary vector

Wolbachia replication and host cell division in Aedes albopictus

Wolbachia pipientis is an obligate intracellular endosymbiont of a range of arthropod species. The microbe is best known for its manipulations of host reproduction that include inducing cytoplasmic incompatibility, parthenogenesis, feminization, and male-killing. Like other vertically transmitted intracellular symbionts, Wolbachiarsquos replication rate must not outpace that of its host cells if it is to remain benign. The mosquito Aedes albopictus is naturally infected both singly and doubly with different strains of Wolbachia pipientis. During diapause in mosquito eggs, no host cell division is believed to occur. Further development is triggered only by subsequent exposure of the egg to water. This study uses diapause in Wolbachia-infected Aedes albopictus eggs to determine whether symbiont replication slows or stops when host cell division ceases or whether it continues at a low but constant rate. We have shown that Wolbachia densities in eggs are greatest during embryonation and then decline throughout diapause, suggesting that Wolbachia replication is dependent on host cell replication.

Host age effect and expression of cytoplasmic incompatibility in field populations of Wolbachia-superinfected Aedes albopictus

The Asian tiger mosquito, Aedes albopictus (Skuse), is a known vector of dengue in South America and Southeast Asia. It is naturally superinfected with two strains of Wolbachia endosymbiont that are able to induce cytoplasmic incompatibility (CI). In this paper, we report the strength of CI expression in crosses involving field-caught males. CI expression was found to be very strong in all crosses between field males and laboratory-reared uninfected or wAlbA infected young females. In addition, crossing experiments with laboratory colonies showed that aged super- infected males could express strong CI when mated with young uninfected or wAlbA infected females. These results provide additional evidence that the CI properties of Wolbachia infecting Aedes albopictus are well suited for applied strategies that seek to utilise Wolbachia for host population modification.

Maternal transmission efficiency of Wolbachia superinfections in Aedes albopictus populations in Thailand

We examined the transmission efficiency of 2 strains of Wolbachia bacteria that cause cytoplasmic incompatibility in field populations of Aedes albopictus by polymerase chain reaction assay. We found mainland and island populations throughout Thailand to be superinfected with group A and B bacteria. Of 320 Wolbachia-positive adult mosquitoes, 97.5% were infected with both groups. Single infected individuals of each Wolbachia group were encountered in nearly equal numbers. We screened 550 offspring from 80 field-collected mothers and found the transmission efficiency of group A Wolbachia to be 96.7% and that of group B Wolbachia to be 99.6%. Mothers that did not transmit both Wolbachia infections to all of their offspring were significantly larger in size than those with perfect transmission fidelity. We discuss our findings in relation to the prospects of the use of Wolbachia as a gene-driving mechanism.

In vitro cultivation of Wolbachia pipientis in an Aedes albopictus cell line

A continuous cell line, Aa23, was established from eggs of a strain of the Asian tiger mosquito, Aedes albopictus, naturally infected with the intracellular symbiont Wolbachia pipientis. The resulting cell line was shown to be persistently infected with the bacterial endosymbiont. Treatment with antibiotics cured the cells of the infection. In the course of establishing this cell line it was noticed that RFLPs in the PCR products of two Wolbachia genes from the parental mosquitoes were fixed in the infected cell line. This indicates that the mosquito host was naturally superinfected with different Wolbachia strains, whereas the infected cell line derived from these mosquitoes only contained one of the original Wolbachia strains. The development of anin vitroculture system for this fastidious microorganism should facilitate molecular analysis of the reproduction distorting phenotypes it induces in natural arthropod hosts.

Wolbachia pipientis: bacterial density and unidirectional cytoplasmic incompatibility between infected populations of Aedes albopictus

Unidirectional cytoplasmic incompatibility is seen when certain Wolbachia-infected insect populations are crossed. Two hypotheses might explain this phenomenon: superinfections with mutually incompatible strains of Wolbachia producing incompatibility when crossed to individuals infected with only a single bacterial strain or, alternatively, a bacterial dosage model, with differences in Wolbachia densities responsible for the incompatibility. A quantitative PCR assay was set up as a general method to compare Wolbachia densities between populations. Using this assay in unidirectionally incompatible stocks of the mosquito Aedes albopictus, we have determined that densities are significantly higher in Houston than in the Mauritius and Koh Samui stocks. This is consistent with a dosage model for the observed crossing patterns, but does not rule out the possibility that superinfection is the primary cause of the incompatibility.