Tecnologias integradas para controle biológico, mecânico e genético de Aedes aegypti / Integrated technologies for biological, mechanical and genetic control of Aedes aegypti

Duas tecnologias alternativas para o controle de Aedes aegypti foram avaliadas: a aplicação espacial de larvicida biológico-Bti em potenciais criadouros peridomiciliares, e a liberação de machos estéreis para inviabilização reprodutiva das fêmeas do mosquito. As ações foram realizadas pelos Agentes dos Serviços de Saúde em 15 vilas da Ilha de Fernando de Noronha, e em uma área (900 imóveis) no bairro da Várzea/Recife/Pernambuco. A efetividade dos métodos foi avaliada por indicadores entomológicos,estimados pela presença, quantidade e viabilidade de ovos do mosquito, coletados em armadilhas, e por marcadores genéticos. A aplicação de Bti, com atomizador costal, ocorreu a cada 30 dias em ambas as áreas. Uma redução importante e sustentável da população de A. aegypti,por este método, foi alcançada em 2015/2016 na Várzea e, em 2016, na Ilha, onde a remoção de 18 toneladas de resíduos sólidos em 2015 contribuiu possivelmente para os resultados. Machos esterilizados com radiação gama foram produzidos em massa no laboratório e liberados em uma das vilas da Ilha. A análise espaço-temporal dos indicadores, de dez/2015a ago/2016, revelou redução expressiva da densidade populacional do mosquito e da diversidade genética da população local. Ambas as abordagens parecem ter reduzido o contato homem-vetor e os riscos de transmissão de arboviroses na Ilha, apesar da elevada competência vetorial da população local do mosquito para os vírus Zika e Dengue. Os métodos testados se mostraram eficientes e passíveis de serem integradas às ações do SUS voltadas ao controle de A. aegypti.

Two alternative technologies were evaluated for Aedes aegypti control:the spraying of a biological larvicide (Bti) in potential peridomiciliarybreeding sites and the release of sterile males to promote reproductionblockage in wild females. Actions were carried out by Agents of theHealth Services, in 15 villages of the Fernando de Noronha Island and in 900 properties from the district of Várzea, Recife-PE. The effectiveness of both methods was evaluated by entomological indicators, estimatedby the presence, quantity and viability of eggs from the mosquito collectedin traps and through genetic markers. Bti was delivered by backpacksprayer every 30 days in both areas. A significant and sustainablereduction of the A. aegypti population as a result of this technique wasachieved in 2015/2016 in Várzea and in 2016 in the Island, where it wasstrengthened by the removal of 18 tons of solid waste in 2015. Malessterilized with gamma radiation were mass-produced in the laboratoryand released in one village of the Island. The spatiotemporal analysis ofthe indicators, from Dec/2015 to Aug/2016, revealed a significant reductionin mosquito density, which impacted on the genetic diversity of thelocal population. Both approaches seem to have reduced human-vectorcontact and the risk of arbovirus transmission in the Island, althoughlocal mosquito population presented high vector competence to Zikaand Dengue virus. These methods were efficient and could be integratedinto SUS actions directed to A. aegypti control.

Monitoring temporal fluctuations of Culex quinquefasciatus using oviposition traps containing attractant and larvicide in an urban environment in Recife, Brazil

The use of attractants and larvicides in oviposition traps is of practical interest for the surveillance and control of urban mosquitoes. In addition to increasing the safety of the traps, this combination is essential for an attract-and-kill control strategy based on trapping mosquito eggs. The combination of Bacillus thuringiensis var. israelensis (Bti) and grass infusion (GI) vs. GI alone were tested for their ability to attract in paired BR-OVT traps in the backyards of 10 houses in Recife, Brazil, for a period of 45 days. Results show that females prefer to oviposit in traps containing Bti (363 compared with 251 egg rafts over 45 days). Results from a one-year trial on the efficacy of BR-OVT traps loaded with GI and Bti as a sampling tool to monitor temporal fluctuations in the population densities of Culex quinquefasciatus in an urban environment are also reported. From December 2006-January 2007, one trap per home was installed and maintained for 348 consecutive days in 134-151 houses located in three urban blocks. Throughout the one-year field trial a total of 43,151 Culex egg rafts were collected in the traps. The data show that BR-OVT loaded with GI and Bti is sensitive enough to demonstrate continuous reproductive activity of Cux. quinquefasciatus in the study area throughout the year and to monitor temporal fluctuations in population density.

An entomological surveillance system based on open spatial information for participative dengue control

Aedes aegypti is a very efficient disseminator of human pathogens. This condition is the result of evolutionary adaptations to frequent haematophagy, as well as to the colonization of countless types of habitats associated with environmental and cultural factors that favor the proliferation of this mosquito in urban ecosystems. Studies using sensitive methods of monitoring demonstrate that the methods of surveillance used in the Brazilian program do not show the high degrees of the infestation of cities by this vector. To increase the capacity of the health sector, new tools are needed to the practice of surveillance, which incorporate aspects of the vector, place and human population. We describe here the SMCP-Aedes - Monitoring System and Population Control of Aedes aegypti, aiming to provide an entomological surveillance framework as a basis for epidemiological surveillance of dengue. The SMCP-Aedes is uphold in the space technology information, supported by the intensive use of the web and free software to collect, store, analyze and disseminate information on the spatial-temporal distribution of the estimated density for the population of Aedes, based on data systematically collected with the use of ovitraps. Planned control interventions, intensified where and when indicated by the entomological surveillance, are agreed with the communities, relying on the permanent social mobilization.

Associadas a fatores bióticos, climáticos e culturais que favorecem a proliferação do Aedes aegypti em ecossistemas urbanos, adaptações evolutivas à hematofagia freqüente e quase exclusiva em humanos e à colonização de tipos infinitos de habitats, fazem deste mosquito um disseminador extremamente eficiente de patógenos ao homem. Estudos utilizando métodos sensíveis de monitoramento demonstram que os métodos de vigilância usados no programa brasileiro não revelam as elevadas intensidades da infestação das cidades por este vetor. Para ampliar a capacidade do setor de saúde novos instrumentos são necessários à prática da vigilância, incorporando aspectos do vetor, do lugar e das pessoas do lugar. Apresentamos aqui o SMCP-Aedes - Sistema de Monitoramento e Controle Populacional do Ae. aegypti, cuja meta é a instrumentalização da vigilância entomológica como base para a vigilância epidemiológica da dengue. Para isso ele se apóia em tecnologias da informação espacial baseadas no uso intensivo da web e de software livre para coletar, armazenar, analisar e disseminar informações relativas à distribuição espaço-temporal da densidade estimada para a população do Aedes, com base em amostras obtidas continuamente com ovitrampas. Intervenções de controle planejadas e intensificadas onde e quando indicado pela vigilância entomológica, são pactuadas com os habitantes, apoiando-se na mobilização social permanente.

Ação mutagênica do inseticida organofosforado temefós em células de medula óssea de camundongos / Mutagenic effects of the organophosphate insecticide temephos on mice bone marrow cells

Foram investigados os efeitos mutagênicos do organofosforado temefós através da observação da formação de micronúcleos em eritrócitos policromáticos (PCEMN) da medula óssea de camundongos. Em camundongos Swiss Webster de ambos os sexos, foram administradas diferentes doses de temefós (27,75; 55,5 e 111,0 mg/kg) por via bucal-gástrica e água destilada (10 mL/kg) no grupo controle negativo; e ciclofosfamida (CPA) (25 mg/kg) foi administrada pela via i.p. no grupo controle positivo. Foram analisadas dez mil células da medula óssea por grupo experimental. Os efeitos mutagênicos foram avaliados nos períodos de 24, 48 e 72h após ministrar a dose única e após nove doses de 111,0 mg/kg (1/semana). Em 24h após a dose única de CPA, a formação de PCEMN foi observada em 1,63% dos camundongos machos e 2,77% em fêmeas. Não houve formação de PCEMN no grupo controle negativo. O temefós induziu PCEMN em 2,61%, 3,50% e 3,69% de animais machos e 1,02%, 1,37% e 1,33% em fêmeas. Após 72h, CPA induziu PCEMN em 0,05% de camundongos de ambos sexos e o temefós em 0,92% de machos e 0,18% em fêmeas. Após nove doses de CPA, houve a formação de PCEMN em 0,15% de machos e em 0,8% de fêmeas; para o temefós, os valores observados foram respectivamente de 0,53% e 0,11%. A ação mutagênica de temefós foi demonstrada pela indução de micronúcleos em camundongos de ambos sexos.

Developing new approaches for detecting and preventing Aedes aegypti population outbreaks: basis for surveillance, alert and control system

A new approach to dengue vector surveillance based on permanent egg-collection using a modified ovitrap and Bacillus thuringiensis israelensis(Bti) was evaluated in different urban landscapes in Recife, Northeast Brazil. From April 2004 to April 2005, 13 egg-collection cycles of four weeks were carried out. Geo-referenced ovitraps containing grass infusion, Bti and three paddles were placed at fixed sampling stations distributed over five selected sites. Continuous egg-collections yielded more than four million eggs laid into 464 sentinel-ovitraps over one year. The overall positive ovitrap index was 98.5 percent (over 5,616 trap observations). The egg density index ranged from 100 to 2,500 eggs per trap-cycle, indicating a wide spread and high density of Aedes aegypti (Diptera: Culicidae) breeding populations in all sites. Fluctuations in population density over time were observed, particularly a marked increase from January on, or later, according to site. Massive egg-collection carried out at one of the sites prevented such a population outbreak. At intra-site level, egg counts made it possible to identify spots where the vector population is consistently concentrated over the time, pinpointing areas that should be considered high priority for control activities. The results indicate that these could be promising strategies for detecting and preventing Ae. aegypti population outbreaks.

Laboratory and field evaluation of an oviposition trap for Culex quinquefasciatus(Diptera: Culicidae)

An ovitrap (BR-OVT) based on physical and chemical stimuli for attracting gravid Culex quinquefasciatus (Diptera: Culicidae) females was developed and evaluated under laboratory and field conditions. Attractants were assayed using alternative chamber bioassays prior to being used in the BR-OVT oviposition trap. A significant preference of gravid females for sites containing conspecific egg rafts was observed, as a response to the natural oviposition pheromone, as well as for sites treated with the synthetic pheromone erythro-6-acetoxy-5-hexadecanolide. Five- to 20-day old grass infusion was strongly attractive to gravid females for laying eggs. On the other hand, entomopathogenic Bacillus sphaericus (Bs) did not influence the choice of an oviposition site when used in combination with grass infusion and can therefore be used as a larvicide in ovitraps. Results from field trials showed that the BR-OVT with grass infusion and with or without Bs works as a preferred oviposition site for Cx. quinquefasciatus. The BR-OVT was more effective for egg collection when placed indoors and comparison with the number of egg rafts laid in cesspits over 40 days indicates that this very simple ovitrap may be a useful tool for monitoring populations of the most important of the vectors of bancroftian filariasis.

Development of a Bacillus sphaericus tablet formulation and its evaluation as a larvicide in the biological control of Culex quinquefasciatus

This study aimed to analyze the final fermentation culture of Bacillus sphaericus 2362, standardize it and develop an active tablet formulation for use in urban mosquito breeding sites. It was performed in three phases: analysis and standardization of a B. sphaericus fermented culture; physical, chemical, and biological analysis of the active powder (solubility, residual humidity, particle size, resting angle, flowing off time, compacted density, and biological activity against Culex quinquefasciatus larvae); and the development of fast-disintegrating tablets. Five formulations with differing compositions were developed and a UV protector was added to the selected formulation. The formulation products with or without UV protector, as well as the active powder caused 100 percent larval mortality from 1 day to 2 months after a single treatment under simulated field conditions. These results show that the UV protector does not affect the initial larvicide activity of B. sphaericus, nor its persistence over a period of two months.

Prevalência da filariose canina causada por Dirofilaria immitis e Dipetalonema reconditum em Maceió, Alagoas, Brasil / Prevalence of canine filiariasis by Dirofilaria immitis and Dipetalonema e reconditum in Maceió, Alagoas State, Brazil

A prevalência da filariose canina causada por Dirofilaria immitis e Dipetalonema reconditum, foi estimada após a realizaçäo de inquérito hemoscópico em 1.519 cäes domiciliados. Foram examinados 1.097 cäes nos sete distritos sanitários da cidade de Maceió, Alagoas, 204 na Ilha de Santa Rita e 218 na cidade de Paripueira. Em Maceió, foram detectados 15 (1,3 por cento) cäes com microfilárias (MF) de D.immitis e 15 (1,3 por cento) com MF de D.reconditum. Todos os casos positivos foram considerados autóctones. As MF de D.immitis e de D. reconditum mediram 298,1µm e 249,2µm de comprimento e 7,3µm e 4,4µm de largura, respectivamente. Além do diagnóstico parasitológico pela técnica de gota espessa (100µL), foi utilizado o teste imunológico Witness, para detecçäo de antígeno filarial circulante, o qual confirmou os dados parasitológicos e permitiu a detecçäo de casos de infecçäo oculta. De um total de 6.579 fêmeas examinadas, pertencentes a seis espécies de mosquitos, oito (0,1 por cento) fêmeas de Culex quinquefasciatus foram encontradas naturalmente infectadas com larvas de D.immitis. Estes resultados demonstram a presença simultânea de D.immitis e D. reconditum na mesma área geográfica.

Evaluation of a new tablet formulation based on Bacillus thuringiensis sorovar. israelensis for larvicidal control of Aedes aegypti

The effect of sunlight on the efficacy and persistence of an experimental tablet formulation based on Bacillus thuringiensis sorovar. israelensis (C4P1) was evaluated against Aedes aegypti larvae under simulated field conditions. The initial mortality ranged from 93 to 100 percent, and the residual activity ( > or = 70 percent mortality) recorded in containers exposed to sunlight or shade were, respectively, 13-35 days and 40-54 days. The results suggest that C4P1 can provide long-term larvicidal effect and operational advantages

The use of bacterial larvicides in mosquite and black fly control programmes in Brazil

Bacillus spp. based larvicides are increasingly replacing, with numerous advantages, chemical insecticides in programmes for controlling black fly and mosquito populations. Brazil was among the pioneers in adopting Bacillus thuringiensis israelensis (B.t.i) to control black flies. However, the major current mosquito control programme in Brazil, the Programme for Eradication of Aedes aegypti launched in 1997, only recently decided to replace temephos by B.t.i based larvicides, in the State of Rio de Janeiro. In the last decade, works developed by research groups in Brazilian institutions have generated a significant contribution to this subject through the isolation of B. sphaericus new strains, the development of new products and the implementation of field trials of Bacillus efficacy against mosquito species under different environmental conditions

Bloodmeal microfilariae density and the uptake and establishment of Wuchereria bancrofti infections in Culex quinquefasciatus and Aedes aegypti

The relationship between ingestion of microfilariae (mf), production of infective larvae (L3) and mf density in human blood has been suggested as an important determinant in the transmission dynamics of lymphatic filariasis. Here we assess the role of these factors in determining the competence of a natural vector Culex quinquefasciatus and a non vector Aedes aegypti to transmit Wuchereria bancrofti. Mosquitoes were infected via a membrane feeding procedure. Both mosquito species ingested more than the expected number of microfilariae (concentrating factor was 1.28 and 1.81 for Cx. quinquefasciatus and Ae. aegypti, respectively) but Cx. quinquefasciatus ingested around twice as many mf as Ae. aegypti because its larger blood meal size. Ae. aegypti showed a faster mf migration capacity compared to Cx. quinquefasciatus but did not allow parasite maturation under our experimental conditions. Similar proportions of melanized parasites were observed in Ae. aegypti (2.4 per cent) and Cx. quinquefasciatus (2.1). However, no relationship between rate of infection and melanization was observed. We conclude that in these conditions physiological factors governing parasite development in the thorax may be more important in limiting vectorial competence than the density of mf ingested.

Controle integrado do vetor da filariose com participaçäo comunitária, em uma área urbana do Recife, Brasil / Integrated control of the filariasis vector with community participation in an urban area of Recife, Pernambuco, Brazil

Um projeto piloto para o controle da filariose bancroftiana foi desenvolvido em duas áreas do Recife, com índices iniciais de prevalência de microfilaremia de 10 por cento. Nas duas áreas, foi aplicado um tratamento em massa com doses baixas de dietilcarbamazina. Em uma das áreas, esse tratamento foi associado ao controle do vetor, integrando medidas físicas, como o uso de camadas flutuantes de esferas de poliestireno, ao tratamento periódico dos criadouros com a bactéria entomopatógena Bacillus sphaericus. Como resultado, a densidade populacional do vetor, de ordem de sessenta a 120 Culex/quarto/noite antes das intervençöes, foi drasticamente reduzida, mantendo-se, durante dois anos, em valores entre quatro e 16 Culex/quarto/noite. Neste contexto, as açöes para envolver a comunidade escolar local no controle do vetor resultaram na realizaçäo de atividades curriculares e extraclasses, culminando com a participaçäo efetiva de grupos de alunos ("Vigilantes de vetores") na aplicaçäo das medidas de controle e na divulgaçäo de conhecimentos básicos sobre a biologia do Culex quinquefasciatus. O entusiástico envolvimento de professores e alunos mostrou ser a escola um espaço aberto à participaçäo em programas desta natureza.

Epidemiological study of brancroftian filariasis in Recife, Northeastern Brazil

Wuchereria bancrofti in Pernambuco was first documented in 1952 (Azevedo & Dobbin 1952), and since then it has been reported in surveys carried out in selected areas of Recife. Several surveys were carried out from 1981 to 1991 by SUCAM. In the 1985 SUCAM's report the disease is considered under control. The CPqAM Filariasis Research Program was established in 1985 and a filarial survey was carried out in the town of Olinda, Greater Recife. In order to verify the real epidemiological situation, a study was conducted in the city of Recife. 21/36 of the Special Zones of Social Interest (ZEIS), were randomly selected for the present study. From 10,664 persons screened, 683 were positive and the prevalence rate for microfilaraemia (mf) varied from 0.6 per cent to 14.9 per cent. A mean mf prevalence of 6.5 per cent, showed that the infection occurs in a wide geographic distribution in Greater Recife and that the intensity of transmission is a real and potential threat to public health in affected communities. Mf rate among males and females differed significantly. Due to the rapid increase in population, unplanned urban settlements, poor sanitary facilities and the favorable geographical conditions to the development of the vector, filariasis may actually be increasing in Recife.

Integrated control measures against Culex quinquefasciatus, the vector of filariasis in Recife

Integrated control measures against Culex quinquefasciastus have been implemented in a pilot urban area in Recife, Brazil. About 3,000 breeding sites found within the operational area were responsible for very high mosquito densities recorded during the pretrial period. Physical control measures have been applied to cess pits before starting a series of 37 treatments of the other sites with Bacillus sphaericus strain 2362, over 27 months. In spite of the difficulties due to environmental conditions, very significant reductions in preimaginal population of C. quinquefasciatus were achieved and, as a consequence, low adult mosquito densities were maintained for a relatively long period of time. Entomological and environmental data gathered in this pilot project can contribute to design an integrated mosquito control program in Recife city.

The reproductive capacity of Panstrongylus megistus (Hemiptera: Reduviidae) males treated with precocenes

Reproductive performance including mating, semen transfer, egg production and hatching rate were investigated in pairs of Panstrongylus megistus in wich the male was treated with either precocene II or ethoxyprecocene II, topically applied shortly after imaginal molt. Eighty percent of the males were not able to mate for a period as long as 40 days after treatment. Those males wich did mate (20%) produced small spermatophores; a few (10%) were unable to transfer spermatozoa to female, while the others (10%) exhibited reproductive performance similar to that of non-treated males. The cumulative effects of the precocene on the different stages of reproduction resulted in a drastic reduction of offspring production. The anti-allatal compound precocene seems to inhibit successful reproduction through a corpus allatum inactivation and subsequent lack of secretory activity of the male accessory glands

Secretory activity and endocrine regulation of male accessory glands in the blood-sucking bug Panstrongylus megistus (Hemiptera: Reduviidae)

The epithelial cells of Panstrongylus megistus male accessory glands (MAG) present ultrastructural characteristics of a secretory cell. Their secretory products are accumulated in the lumen of the four MAG lobes. During the first 8 days of adult life a strong secretion activity occurs, accumulating enough material to produce the first spermatophore. Cerebral neurosecretions as well as juvenile hormone are both involved in MAG secretory activity regulation. Juvenile hormone seems to be the responsible for the stimulation of most protein synthesis in male accessory glands. Cerebral neurosecretion seems to be necessary to stimulate juvenile hormone production and release by the corpus allatum. Furthermore, neurosecretion is required for some polypeptides synthesis by MAG. Although topic application of precocene II to adult males does not reproduce the same effects on MAG as does allatectomy, this compound causes strong reduction on male reproductive capacity.