Genetic Control of Mosquitoes: population suppression strategies

Over the last two decades, morbidity and mortality from malaria and dengue fever among other pathogens are an increasing Public Health problem. The increase in the geographic distribution of vectors is accompanied by the emergence of viruses and diseases in new areas. There are insufficient specific therapeutic drugs available and there are no reliable vaccines for malaria or dengue, although some progress has been achieved, there is still a long way between its development and actual field use. Most mosquito control measures have failed to achieve their goals, mostly because of the mosquito's great reproductive capacity and genomic flexibility. Chemical control is increasingly restricted due to potential human toxicity, mortality in no target organisms, insecticide resistance, and other environmental impacts. Other strategies for mosquito control are desperately needed. The Sterile Insect Technique (SIT) is a species-specific and environmentally benign method for insect population suppression, it is based on mass rearing, radiation mediated sterilization, and release of a large number of male insects. Releasing of Insects carrying a dominant lethal gene (RIDL) offers a solution to many of the drawbacks of traditional SIT that have limited its application in mosquitoes while maintaining its environmentally friendly and species-specific utility. The self-limiting nature of sterile mosquitoes tends to make the issues related to field use of these somewhat less challenging than for self-spreading systems characteristic of population replacement strategies. They also are closer to field use, so might be appropriate to consider first. The prospect of genetic control methods against mosquito vectored human diseases is rapidly becoming a reality, many decisions will need to be made on a national, regional and international level regarding the biosafety, social, cultural and ethical aspects of the use and deployment of these vector control methods.

EVALUATION OF ANTIMICROBIAL AND CYTOTOXIC ACTIVITIES OF PLANT EXTRACTS FROM SOUTHERN MINAS GERAIS CERRADO

The antimicrobial activity of plant hidroethanolic extracts on bacteria Gram positive, Gram negative, yeasts, Mycobacterium tuberculosis H37 and Mycobacterium bovis was evaluated by using the technique of Agar diffusion and microdilution in broth. Among the extracts evaluated by Agar diffusion, the extract of Bidens pilosa leaf presented the most expressive average of haloes of growth inhibition to the microorganisms, followed by the extract of B. pilosa flower, of Eugenia pyriformis' leaf and seed, of Plinia cauliflora leaf which statistically presented the same average of haloes inhibitory formation on bacteria Gram positive, Gram negative and yeasts. The extracts of Heliconia rostrata did not present activity. Mycobacterium tuberculosis H37 and Mycobacterium bovis(BCG) appeared resistant to all the extracts. The susceptibility profile of Candida albicans and Saccharomyces cerevisiae fungi were compared to one another and to the Gram positive Bacillus subtilis, Enterococcus faecalis and the Gram negative Salmonella typhimurium bacteria (p > 0.05). The evaluation of cytotoxicity was carried out on C6-36 larvae cells of the Aedes albopictus mosquito. The extracts of stem and flower of Heliconia rostrata, leaf and stem of Plinia cauliflora, seed of Anonna crassiflora and stem, flower and root of B. pilosa did not present toxicity in the analyzed concentrations. The highest rates of selectivity appeared in the extracts of stem of A. crassiflora and flower of B. pilosa to Staphylococcus aureus, presenting potential for future studies about a new drug development.

SAINT LOUIS ENCEPHALITIS VIRUS IN MATO GROSSO, CENTRAL-WESTERN BRAZIL

The dengue virus (DENV), which is frequently involved in large epidemics, and the yellow fever virus (YFV), which is responsible for sporadic sylvatic outbreaks, are considered the most important flaviviruses circulating in Brazil. Because of that, laboratorial diagnosis of acute undifferentiated febrile illness during epidemic periods is frequently directed towards these viruses, which may eventually hinder the detection of other circulating flaviviruses, including the Saint Louis encephalitis virus (SLEV), which is widely dispersed across the Americas. The aim of this study was to conduct a molecular investigation of 11 flaviviruses using 604 serum samples obtained from patients during a large dengue fever outbreak in the state of Mato Grosso (MT) between 2011 and 2012. Simultaneously, 3,433 female Culex spp. collected with Nasci aspirators in the city of Cuiabá, MT, in 2013, and allocated to 409 pools containing 1-10 mosquitoes, were also tested by multiplex semi-nested reverse transcription PCR for the same flaviviruses. SLEV was detected in three patients co-infected with DENV-4 from the cities of Cuiabá and Várzea Grande. One of them was a triple co-infection with DENV-1. None of them mentioned recent travel or access to sylvatic/rural regions, indicating that transmission might have occurred within the metropolitan area. Regarding mosquito samples, one pool containing one Culex quinquefasciatus female was positive for SLEV, with a minimum infection rate (MIR) of 0.29 per 1000 specimens of this species. Phylogenetic analysis indicates both human and mosquito SLEV cluster, with isolates from genotype V-A obtained from animals in the Amazon region, in the state of Pará. This is the first report of SLEV molecular identification in MT.

INVENTORY OF MOSQUITOES (DIPTERA: CULICIDAE) IN CONSERVATION UNITS IN BRAZILIAN TROPICAL DRY FORESTS

In Brazil, most studies of the Culicidae family are concentrated in rainforest regions. As such, there is a lack of knowledge regarding the diversity of Culicidae in regions with different climatic and vegetational characteristics. The aim of this study was to compile an inventory of Culicidae in protected areas of the semi-arid region of the state of Minas Gerais, Brazil, in order to better understand the diversity of the family within this region. The study was conducted across four protected areas in the northern region of the state, in tropical dry forest (TDF) fragments. Sampling methods included Shannon trap and CDC light trap, as well as active collection. A total of 11,219 mosquito specimens were collected between August 2008 and July 2012, belonging to 11 genera and 45 species; 15 new records for the state of Minas Gerais were registered, as well as 26 new records for semi-arid regions within the state. The high number of new Culicidae records in this region demonstrates the importance of inventory studies for increasing the knowledge of culicid biodiversity in Minas Gerais, and in particular within semi-arid regions of the state.

SEASONAL DISTRIBUTION OF MALARIA VECTORS (DIPTERA: CULICIDAE) IN RURAL LOCALITIES OF PORTO VELHO, RONDÔNIA, BRAZILIAN AMAZON

We conducted a survey of the malaria vectors in an area where a power line had been constructed, between the municipalities of Porto Velho and Rio Branco, in the states of Rondônia and Acre, respectively. The present paper relates to the results of the survey of Anopheles fauna conducted in the state of Rondônia. Mosquito field collections were performed in six villages along the federal highway BR 364 in the municipality of Porto Velho, namely Porto Velho, Jaci Paraná, Mutum Paraná, Vila Abunã, Vista Alegre do Abunã, and Extrema. Mosquito captures were performed at three distinct sites in each locality during the months of February, July, and October 2011 using a protected human-landing catch method; outdoor and indoor captures were conducted simultaneously at each site for six hours. In the six sampled areas, we captured 2,185 mosquitoes belonging to seven Anopheles species. Of these specimens, 95.1% consisted of Anopheles darlingi, 1.8% An. triannulatus l.s., 1.7% An. deaneorum, 0.8% An. konderi l.s., 0.4 An. braziliensis, 0.1% An. albitarsis l.s., and 0.1% An. benarrochi. An. darlingi was the only species found in all localities; the remaining species occurred in sites with specific characteristics.

VIROLOGICAL AND SEROLOGICAL DIAGNOSIS OF RABIES IN BATS FROM AN URBAN AREA IN THE BRAZILIAN AMAZON

The outbreaks of rabies in humans transmitted by Desmodus rotundus in 2004 and 2005, in the northeast of the Brazilian State of Para, eastern Amazon basin, made this a priority area for studies on this zoonosis. Given this, the present study provides data on this phenomenon in an urban context, in order to assess the possible circulation of the classic rabies virus (RABV) among bat species in Capanema, a town in the Amazon basin. Bats were collected, in 2011, with mist nets during the wet and dry seasons. Samples of brain tissue and blood were collected for virological and serological survey, respectively. None of the 153 brain tissue samples analyzed tested positive for RABV infection, but 50.34% (95% CI: 45.67-55.01%) of the serum samples analyzed were seropositive. Artibeus planirostris was the most common species, with a high percentage of seropositive individuals (52.46%, 95% CI: 52.31 52.60%). Statistically, equal proportions of seropositive results were obtained in the rainy and dry seasons (c2 = 0.057, d.f. = 1, p = 0.88). Significantly higher proportions of males (55.96%, 95% CI: 48.96-62.96%) and adults (52.37%, 95% CI: 47.35-57.39%) were seropositive. While none of the brain tissue samples tested positive for infection, the high proportion of seropositive specimens indicates that RABV may be widespread in this urban area.

EVALUATION OF SUBLETHAL EFFECTS OF Ipomoea cairica LINN. EXTRACT ON LIFE HISTORY TRAITS OF DENGUE VECTORS

Plant derived insecticides have considerable potential for mosquito control because these products are safer than conventional insecticides. This study aimed to investigate sublethal activities of Ipomoea carica or railway creeper crude acethonilic extract against life history trait of dengue vectors, Aedes albopictus and Aedes aegypti. The late third instar larvae of Ae. albopictus and Ae. aegypti were exposed to a sublethal dose at LC50 and larvae that survived were further cultured. Overall, Ipomea cairica crude extracts affected the whole life history of both Aedes species. The study demonstrated significantly lower egg production (fecundity) and eggs hatchability (fertility) in Ae. albopictus. The sublethal dose of crude extracts reduced significantly the width of larval head capsule and the wing length of both sexes in both Aedes species. The significance of sublethal effects of I. cairica against Aedes mosquitoes was an additional hallmark to demonstrate further activity of this plant despite its direct toxicity to the larvae. The reduced reproductive capacity as well as morphological and physiological anomalies are some of the effects that make I. cairica a potential candidate to be used as a new plant-based insecticide to control dengue vectors.

Symptomless Plasmodium vivax parasitemias and malaria eradication in Santa Catarina State, Brazil

Thirty-nine symptomless carriers of P. vivax parasites in the blood gave blood films at monthly intervals for four to six months during the non- transmission season. It was found that parasitaemias can continue for many months. Thirteen of those studied relapsed with symptoms and were treated with chloroquine at a dosage of 600 mg for adults with proportionate doses for children. Of these nine relapsed silently while under observation: a symptomless relapse rate of approximately 70 per cent. One case had symptoms attributable to malaria close to the time of the original survey (the day before). Of the remaining 38 asymptomatic parasite carriers four showed microgametocytes in a density that suggested a potentially high infectivity and six showed microgametocytes in a density suggesting a potentially low to moderate infectivity for mosquito vectors. There was thus a proportion of one smptomatic case of malaria to 10 potentially infective symptomless parasite carriers. Because they feel no need to seek treatment, such persons may form an important reservoir of infection when vectors cannot be fully controlled by spraying. Some possible methods of dealing with such situations are discussed.

Observações sobre anofelinos em Santa Catarina, Brasil

Um ciclo anual de capturas de mosquitos realizado em três localidades da Região do Litoral e Encosta de Santa Catarina, com um homemisca colocado na mata, outro na casa mais próxima e um terceiro situado entre esses dois pontos, forneceu as seguintes informações: 1) O Anopheles bellator que é um mosquito raro nas matas densas das encostas, torna-se abundante quando essas florestas são danificadas pela retirada de árvores; 2) Tanto essa espécie quanto o A. cruzii e o A. homunculus, compareceram em maior número nas capturas feitas no terreno descampado; 3) A não ser no fato do A. homunculus procurar alimento nas horas um pouco mais avançadas da noite, não se notou diferença significativa entre o comportamento dessas três espécies de anofelinos; 4) A comparação dos dados obtidos, com os de outros autores, mostrou que uma pessoa colocada fora da casa intercepta um número significativo de mosquitos; 5) Ficou bem claro que os anofelinos da região raramente pousam em paredes dedetizadas; 6) Além disso, o DDT diminui muito a proporção de exemplares que vai se alimentar dentro das casas; para o A. cruzii essa diminuição foi da ordem de 90%.

Uso de células de Aedes albopictus C6/36 na propagação e classificação de arbovírus das famílias Togaviridae, Flaviviridae, Bunyaviridae e Rhabdoviridae

Colônias de células de mosquito Aedes albopictus C6/36 foram infectadas com 23 arbovirus, sendo 19 destes existentes no Brasil, pertencentes às famílias Togaviridae, Flaviviridae, Bunyaviridae e Rhabdoviridae. A Replicação virai foi detectada por imunofluorescência indireta com todos os vírus estudados enquanto que o efeito citopático foi observado durante a infecção por alguns destes. No teste de imunofluorescência indireta utilizou-se fluidos ascíticos imunes de camundongos, específicos para os vírus estudados. A replicação virai caracterizada por grande produção de antígeno recomenda a utilização de células C6/36 na propagação e em tentativas de isolamento desses arbovirus. A técnica de imunofluorescência ofereceu importantes subsídios na classificação e identificação de vírus que replicam nestas células.

Biting behavior of Anopheles mosquitoes in Costa Marques, Rondonia, Brazil

Mosquito collections were made in and near Costa Marques, Rondonia, Brazil, to determine anopheline anthropophilic/zoophilic behavior. Collections from a non-illuminated, bovine-baited trap and indoor and outdoor human-bait collections were compared. Anopheles darlingi and Anopheles deaneorum were more anthropophilic than the other anophelines collected. The remainder of the Anopheles species were collected much morefrequently in bovine-baited traps than in human-bait collections. Anopheles darlingi and An. deaneorum were more frequently collected inside houses than the other anopheline species. But, when collections were made in a house with numerous openings in the walls, there were few differences in the percentages of each species biting man indoors versus outdoors. Anopheles darlingi was the predominant mosquito collected, both inside and outside houses, and had the strongest anthropophilic feeding behavior of the anophelines present.

A febre amarela na região de Ribeirão Preto durante a virada do século XIX: importância científica e repercussões econômicas

Nesta revisão histórica, descreve-se a situação precária da saúde pública no Brasil durante o século XIX devido as múltiplas epidemias de febre amarela. Descreve-se, também, os precários conhecimentos que haviam, à época, sobre a enfermidade. Faz- se um breve histórico sobre a Região de Ribeirão Preto, situada no Nordeste do Estado de São Paulo, seus desbravadores, imigrantes e fazendeiros de café pioneiros, incluindo-se Luiz Pereira Barreto. Relatam-se as 3 epidemias de febre amarela ocorridas na cidade de São Simão, nos anos de 1896. 1898, 1902 e uma ocorrida em Ribeirão Preto, no ano de 1903. Mostra-se que as medidas de saúde pública tomadas para o controle dos surtos mostraram-se eficientes quando Emílio Ribas promoveu, de forma pioneira, o combate ao mosquito transmissor, o Aedes aegypti. Descrevem-se os experimentos efetuados por Emílio Ribas, com a participação de Adolpho Lutz e de Luiz Pereira Barreto, visando confirmar a transmissão vetorial da febre amarela e afastar outras formas de contágio. Obseva-se que as epidemias de febre amarela causaram prejuízo ao desenvolvimento da Cidade de São Simão, colaborando para a transferência do polo econômico regional para Ribeirão Preto. Ressalta-se a importância das medidas de controle vetorial e a dos experimentos sobre o mecanismo de transmissão da febre amarela, no desenvolvimento da ciência médica, e da saúde pública no Brasil.

Descrição da colonização de Aedes albopictus (Diptera: culicidae) na região de São José do Rio Preto, SP, 1991-1994

O objetivo deste trabalho é descrever e analisar colonização do Aedes albopictus, cuja presença foi detectada na região de São José do Rio Preto em 1991, jã colonizada pelo Aedes aegypti. A partir de informações obtidas em medidas de densidade larvária pela Superintendência de Controle de Endemias (SUCEN), analisou-se: ano e mês de oconência, município, composição e localização das amostras lanúrias, tipo de recipiente, número médio de laivas e índice de Breteau. Até dezembro de 1994 a presença do As. albopictus fora constatada em 34 municípios. A colonização da região pelo mosquito ainda é reduzida, apresentando algumas diferenças em relação ao Ae. aegypti: maior proporção no peridomicílio, ocupando recipientes em proporções diferentes. O número médio de larvas de As. albopictus sofreu influência da presença de larvas da outra espécie. Apresentou comportamento sazonal semelhante ao do Ae. aegypti e avançou no sentido leste para oeste.

Descrição da colonização de Aedes aegypti na região de São José do Rio Preto, São Paulo

O objetivo do trabalho é descrever a colonização da região pelo Aedes aegypti. Levantamento entomológico realizado em 1985 detectou a espécie em São José do Rio Preto. A dispersão do mosquito atingiu, até 1988, os 30 municípios da região. Nos distritos e aglomerados rurais, o primeiro foco do vetor foi encontrado em 1987 em um dos 29 existentes, dispersando-se para os demais até 1991. Os focos foram identificados, principalmente, através de pesquisas larvárias em locais com grande concentração de recipientes, e a maior freqüência de encontro de larvas de Ae. aegypti ocorreu em pneus, principais responsáveis por sua dispersão. Os focos foram identificados, basicamente, entre novembro e abril, períodos de maior incidência de chuvas. As delimitações dos focos mostraram que os principais recipientes infestados pelo mosquito nos domicílios foram os pneus e vasos de plantas. A conseqüência mais importante da presença do Ae. aegypti tem sido as ocorrências de epidemias de dengue.

Influência do período de quiescência dos ovos sobre o ciclo de vida de Aedes aegypti (Linnaeus, 1762) (Diptera, Culicidae) em condições de laboratório

Estudou-se a influência do período de quiescência dos ovos no ciclo de vida de Aedes aegypti (Linnaeus, 1762) (Diptera, Culicidae) em condições de laboratório, na busca de informações que possam melhorar o direcionamento das ações de controle, pois sabe-se que o ovo é a forma mais resistente do ciclo biológico, possibilitando ao mosquito ampla sobrevida, devido à resistência às adversidades climáticas. Os experimentos foram realizados numa câmara biológica, mantida à temperatura de 28 ± 1oC, com umidade relativa de 80 ± 5% e fotofase de 12 horas. Apresentam-se os dados da influência de diferentes períodos de quiescência sobre a eclosão das larvas, desenvolvimento larval e pupal, ciclo evolutivo. Verificou-se o efeito altamente significativo do período de quiescência na eclosão das larvas. O período de quiescência não influenciou nas durações dos períodos de incubação, larval e pupal. Constatou-se que ovos de um mesmo período de quiescência apresentaram períodos de incubação estatisticamente diferentes entre si. As larvas eclodiam em grupos, definidos pela incubação, e este efeito de grupo foi significativo na duração do ciclo. Pode-se afirmar que, em 99,8% dos ciclos, a variação foi determinada pela incubação.