Interruption of lymphatic filariasis transmission in Manaus, a former focus of Wuchereria bancrofti in the Western Brazilian Amazon / Interrupción de la transmisión de la filariasis linfática en Manaos, anteriormente un foco de Wuchereria bancrofti en la Amazonia occidental de Brasil / Interrupção da transmissão da filariose linfática em Manaus, anteriormente um foco de Wuchereria bancrofti na parte leste da Amazônia brasileira

ABSTRACT Objective. To confirm the absence of Wuchereria bancrofti autochthonous cases in Manaus, a former focus of lymphatic filariasis in the Western Brazilian Amazon. Methods. A field survey was carried out in 2016 using immunochromatographic rapid tests (ICT card) for the detection of circulating filarial antigens in blood. The sample included a group of 3 000 schoolchildren aged 6 to 10 years enrolled in schools from different urban areas of Manaus (including the former lymphatic filariasis focus in the city) and a group of 709 adolescents and adults, between the ages of 11 and 85 years, born and raised in different areas of Manaus. Results. All of the individuals tested negative for W. bancrofti antigen. Conclusions. Although Manaus was once considered endemic, this focus no longer seems to be active for lymphatic filariasis transmission. The results of this study could support the certification by the World Health Organization of the lymphatic filariasis transmission elimination exercise in Brazil.

RESUMEN Objetivo. Confirmar la ausencia de casos autóctonos de Wuchereria bancrofti en Manaos, anteriormente un foco de filariasis linfática en la Amazonia occidental de Brasil. Métodos. En el 2016 se llevó a cabo una encuesta en el terreno con pruebas rápidas inmunocromatográficas (tiras inmunocromatográficas) para detectar antígenos filáricos circulantes en sangre. La muestra constó de un grupo de 3 000 escolares de 6 a 10 años matriculados en escuelas de diferentes zonas urbanas de Manaos (incluida la zona que anteriormente era el foco de filariasis linfática en la ciudad) y de un grupo de 709 adolescentes y adultos, de edades comprendidas entre 11 y 85 años, nacidos y criados en diferentes áreas de Manaos. Resultados. Todas las personas dieron negativo en la prueba de antígeno de Wuchereria bancrofti. Conclusiones. Aunque hubo un tiempo en que Manaos se consideraba zona endémica, parece que este foco de transmisión de la filariasis linfática ya no está activo. Los resultados de este estudio podrían brindar apoyo a la certificación de la Organización Mundial de la Salud respecto de los esfuerzos realizados en Brasil para eliminar la transmisión de la filariasis linfática.

RESUMO Objetivo. Confirmar a ausência de casos autóctones de Wuchereria bancrofti em Manaus, anteriormente um foco da filariose linfática na parte leste da Amazônia brasileira. Métodos. Uma pesquisa de campo foi realizada em 2016 com o uso de teste rápido por imunocromatografia (cartão ICT) para detecção de antígenos de microfilárias circulantes no sangue. A amostra estudada consistiu de um grupo de 3 000 crianças escolares entre 6 e 10 anos de idade matriculados em escolas de diferentes áreas da zona urbana de Manaus (englobando a área anteriormente com o foco de filariose linfática) e um grupo de 709 adolescentes e adultos entre 11 e 85 anos de idade nascidos e crescidos em diferentes áreas de Manaus. Resultados. Todos os indivíduos pesquisados tiveram teste negativo para o antígeno da W. bancrofti. Conclusões. Apesar de Manaus ter sido anteriormente uma área endêmica, parece que não existe mais foco ativo de transmissão da filariose linfática na cidade. Os resultados deste estudo podem servir para embasar a certificação pela Organização Mundial da Saúde da eliminação da transmissão da filariose linfática no Brasil.

Evaluation of the recombinant antigens Wb14 and WbT for the capture antibody diagnosis of lymphatic filariasis

BACKGROUND Lymphatic filariasis (LF) is a parasitic disease caused mainly by the Wuchereria bancrofti worm and that affects up to 120 million people worldwide. LF is the second cause of chronic global deformity, responsible for 15 million people with lymphedema (elephantiasis) and 25 million men with scrotal hydrocele. Its diagnosis is still associated with numerous difficulties, such as the sample collection periods (microfilaria nocturnal periodicity) and limited diagnostic kits. OBJECTIVES The aim of this work was to evaluate two recombinant antigens (Wb14 and WbT) as part of an enzyme-linked immunosorbent assay (ELISA) based antibody capture tests for LF. METHODS The recombinant antigens rWb14 and rWbT were expressed in Escherichia coli BL21 and an antibody capture ELISA was performed. For this, sera were used from microfilaremic individuals with W. bancrofti (MF), chronic pathology (CP), individuals infected with Strongyloides (SP) and healthy controls from endemic (EN) and non-endemic (NE) areas. FINDINGS Both tests showed similar results, with 90% sensitivity and 96.6% specificity. In comparison with the BM14 ELISA commercial test, the Wb14 and WbT antigens performed with identical sensitivity but greater specificity. Reduced positivity with the CP suggested a potential to monitor cure. This was not confirmed, however, when sera from individuals up to seven years after treatment were assayed. MAIN CONCLUSIONS The Wb14 and WbT ELISAs were considered efficient and promising diagnostic tests. Due to the importance of antibody capture analysis to evaluate the Global Program to Eliminate Lymphatic Filariasis (GPELF), the tests proposed here appear as great alternatives to the available commercial system.

Wuchereria bancrofti infection in Haitian immigrants and the risk of re-emergence of lymphatic filariasis in the Brazilian Amazon

Abstract INTRODUCTION: Lymphatic filariasis (LF) is a public health problem in Haiti. Thus, the emigration of Haitians to Brazil is worrisome because of the risk for LF re-emergence. METHODS: Blood samples of Haitian immigrants, aged ≥18 years, who emigrated to Manaus (Brazilian Amazon), were examined using thick blood smears, membrane blood filtration, and immunochromatography. RESULTS: Of the 244 immigrants evaluated, 1 (0.4%) tested positive for W. bancrofti; 11.5% reported as having received LF treatment in Haiti. CONCLUSIONS: The re-emergence of LF in Manaus is unlikely, due to its low prevalence and low density of microfilaremia among the assessed Haitian immigrants.

Histórico das ações de controle da filariose linfática EM Olinda, Pernambuco, Brasil / History of lymphatic filariasis control actions in Olinda, Pernambuco, Brazil

A filariose linfática é uma parasitose que afeta regiões tropicais e subtropicais. No Brasil, apenas a região metropolitana de Recife (Recife, Olinda, Jaboatão dos Guararapes e Paulista) ainda é considerada como foco ativo de transmissão da parasitose. Diante disso, o objetivo deste trabalho foi descrever o histórico das atividades de controle da parasitose em OlindaPE, área que por muito tempo apresentou número significativo de casos, evidenciando as principais ações desenvolvidas desde o ano de 1987. Ao longo de aproximadamente 30 anos, muitos estudos clínico-laboratoriais e epidemiológicos foram desenvolvidos no município. Recentemente, mais de cinco ciclos de tratamento em massa com citrato de dietilcarbamazina foram realizados na área, com uma adesão de mais de 65% de toda a população, o que contribuiu para a redução da microfilaremia local a taxas inferiores a 1%. Esse fato viabilizou a implantação da pesquisa de avaliação da transmissão, etapa decisiva para definição da situação atual da filariose linfática e de novas estratégias de controle e vigilância.

Mosquito identification and molecular xenomonitoring of lymphatic filariasis in selected endemic areas in Giza and Qualioubiya governorates, Egypt

Lymphatic filariasis is a vector-borne health problem that has been focally endemic in Egypt for centuries. The chief vectors of transmission are Culicinae species. Control measures in the form of mass drug administration of DEC citrate treatment have been implemented in Nile delta for almost a decade. This study aimed to identify the prevalent mosquito species in endemic areas in Giza and Qualioubiya governorates and to monitor Wuchereria bancrofti infection by detecting the parasite DNA in collected mosquitoes. Adult mosquitoes were collected using light traps hung indoors. Microscopic examination was performed to identify and examine the morphologic characters of mosquitoes. Female Culex mosquitoes were subjected to semi-nested PCR to detect filarial DNA targeting repetitive DNA sequences [pWb12 repetitive region] specific for W.bancrofti

The results revealed 3 species of mosquitoes Culex pipiens, Culex pusillus and Culex quinquefasciatus with the predominance of Culex pipiens [85.7%]. Wuchereria bancrofti DNA was not detected in any of the collected mosquito pools. With the progress of elimination programme in Nile Delta, follow up studies with larger sample size are recommended as the predominance of Culex pipiens the main lymphatic filariasis vector remains a risk of transmission in such areas

Finding Wolbachia in Filarial larvae and Culicidae Mosquitoes in Upper Egypt Governorate

Wolbachia is an obligatory intracellular endosymbiotic bacterium, present in over 20% of all insects altering insect reproductive capabilities and in a wide range of filarial worms which is essential for worm survival and reproduction. In Egypt, no available data were found about Wolbachia searching for it in either mosquitoes or filarial worms. Thus, we aimed to identify the possible concurrent presence of Wolbachia within different mosquitoes and filarial parasites, in Assiut Governorate, Egypt using multiplex PCR. Initially, 6 pools were detected positive for Wolbachia by single PCR. The simultaneous detection of Wolbachia and filarial parasites (Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens) by multiplex PCR was spotted in 5 out of 6 pools, with an overall estimated rate of infection (ERI) of 0.24%. Unexpectedly, the highest ERI (0.53%) was for Anopheles pharoensis with related Wolbachia and W. bancrofti, followed by Aedes (0.42%) and Culex (0.26%). We also observed that Wolbachia altered Culex spp. as a primary vector for W. bancrofti to be replaced by Anopheles sp. Wolbachia within filaria-infected mosquitoes in our locality gives a hope to use bacteria as a new control trend simultaneously targeting the vector and filarial parasites.

Avaliação da infecção filarial por wuchereria bancrofti antes do início do tratamento massivo para eliminação da filaríase linfática no distrito de murrupula, província de nampula-Moçambique

A Wuchereria bancrofti é responsável por mais de 90 por cento dos casos da Filaríase Linfática no mundo todo. Em Moçambique, 103 distritos são endémicos para a doença com as prevalências mais altas na região norte do país. A eliminação da doença esta prevista para o ano 2020 e pode ser alcançada pelo tratamento massivo das populações das áreas endémicas durante 5-6 anos com ivermectina e albendazole. A avaliação do impacto do tratamento e a verificação da eliminação da doença carece de ferramentas de diagnóstico com sensibilidade suficiente para detectar os mais pequenos vestígios de infecção remanescente. Entre elas, o teste imunocromatográfico (ICT) para pesquisa do antígeno filarial circulante (AFC), a gota espessa (GE) de sangue para pesquisa de microfilárias (Mf) e a PCR para a detecção do ADN têm sido frequentemente utilizadas antes (baseline), durante e após a campanha de eliminação com vista a monitorar a situação de transmissão da doença. Assim, no presente inquérito de base (baseline) propusemo-nos a avaliar a presença da infecção filarial por Wuchereria bancrofti antes do início do tratamento massivo pelos métodos comummente utilizados (teste imunocromatográfico e gota espessa) e também para o efeito de padronização da técnica, faze-lo utilizando um método de detecção molecular (PCR). As amostras de sangue foram colhidas no período nocturno entre as 21h-2h da manhã por punção digital. O teste imunocromatográfico foi realizado durante o inquérito utilizando 100 microlitro de sangue. As lâminas foram coradas pelo Giemsa e duplamentes observadas ao microscópio óptico para contagem de microfilárias.

Xenomonitoramento molecular para avaliação da infecção vetorial por Wuchereria bancrofti em áreas endêmicas de filariose linfática na Região Metropolitana do Recife/PE / Molecular xenomonitoring for the evaluation of vector infection rate by Wuchereria bancrofti in endemic areas for lymphatic filariasis in Metropolitan Region of Recife/PE

A filariose linfática (FL) ou bancroftiana é uma doença parasitária causada por Wuchereria bancrofti, um verme filarial transmitido no Brasil pelo mosquito Culex quinquefasciatus. De acordo com a Organização Mundial de Saúde (OMS) esta doença afeta 120 milhões de pessoas em 58 países. Portanto, para enfrentar a FL, a OMS lançou um programa global para eliminá-la até 2020 e o Brasil tornou signatário dessa proposta criando o Plano Nacional de Eliminação da Filariose Linfática (PNEFL). Atualmente, a Região Metropolitana do Recife (RMR) é uma área de importante transmissibilidade e, assim, foi preconizado o Tratamento Coletivo (TC) da população com o medicamento Dietilcarbamazina (DEC) e o controle vetorial para reduzir a transmissão da doença. Como ferramenta complementar, desde a vigilância até a verificação da eliminação, o xenomonitoramento molecular (baseado na PCR para detecção de W. bancrofti em mosquitos) é um importante método não invasivo para monitorar indiretamente se a transmissão de larvas de W. bancrofti está ocorrendo na população humana. A fim de verificar a taxa de infecção vetorial no mosquito C. quinquefasciatus pela W. bancrofti foram coletadas 43.981 fêmeas do mosquito em doze localidades na RMR. Além disso, foi desenvolvido um novo protocolo (PCR duplex) para o diagnóstico de infecção vetorial e o número ideal de fêmeas por pools foi estabelecido. Os resultados mostraram que Linha do Tiro (Recife), uma área com alto índice de microfilaremia na população humana, apresentou status de transmissão durante o TC com uma taxa de infecção vetorial de 0,80 por cento, diferente das outras localidades com transmissão reduzida não foram detectados pools positivos. Portanto, observa-se que onde o TC é conduzido a taxa de infecção vetorial tende a ser reduzida. O xenomonitoramento molecular é um indicador importante para avaliação da eficiência das estratégias do PGEFL implantado em áreas endêmicas, até que ocorra a certificação da interrupção do ciclo de transmissão da filariose.

Xenomonitoring of Different Filarial Nematodes Using Single and Multiplex PCR in Mosquitoes from Assiut Governorate, Egypt

Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens are filarial nematodes transmitted by mosquitoes belonging to Culex, Aedes, and Anopheles genera. Screening by vector dissection is a tiresome technique. We aimed to screen filarial parasites in their vectors by single and multiplex PCR and evaluate the usefulness of multiplex PCR as a rapid xenomonitoring and simultaneous differentiation tool, in area where 3 filarial parasites are coexisting. Female mosquitoes were collected from 7 localities in Assiut Governorate, were microscopically identified and divided into pools according to their species and collection site. Detection of W. bancrofti, D. immitis, and D. repens using single PCR was reached followed by multiplex PCR. Usefulness of multiplex PCR was evaluated by testing mosquito pools to know which genera and species are used by filarial parasites as a vector. An overall estimated rate of infection (ERI) in mosquitoes was 0.6%; the highest was Culex spp. (0.47%). W. bancrofti, D. immitis, and D. repens could be simultaneously and differentially detected in infected vectors by using multiplex PCR. Out of 100 mosquito pools, 8 were positive for W. bancrofti (ERI of 0.33%) and 3 pools each were positive for D. immitis and D. repens (ERI 0.12%). The technique showed 100% sensitivity and 98% specificity. El-Nikhila, El-Matiaa villages, and Sahel Seleem district in Assiut Governorate, Egypt are still endemic foci for filarial parasites. Multiplex PCR offers a reliable procedure for molecular xenomonitoring of filariasis within their respective vectors in endemic areas. Therefore, it is recommended for evaluation of mosquito infection after lymphatic filariasis eradication programs.

Xenomonitoring of Different Filarial Nematodes Using Single and Multiplex PCR in Mosquitoes from Assiut Governorate, Egypt

Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens are filarial nematodes transmitted by mosquitoes belonging to Culex, Aedes, and Anopheles genera. Screening by vector dissection is a tiresome technique. We aimed to screen filarial parasites in their vectors by single and multiplex PCR and evaluate the usefulness of multiplex PCR as a rapid xenomonitoring and simultaneous differentiation tool, in area where 3 filarial parasites are coexisting. Female mosquitoes were collected from 7 localities in Assiut Governorate, were microscopically identified and divided into pools according to their species and collection site. Detection of W. bancrofti, D. immitis, and D. repens using single PCR was reached followed by multiplex PCR. Usefulness of multiplex PCR was evaluated by testing mosquito pools to know which genera and species are used by filarial parasites as a vector. An overall estimated rate of infection (ERI) in mosquitoes was 0.6%; the highest was Culex spp. (0.47%). W. bancrofti, D. immitis, and D. repens could be simultaneously and differentially detected in infected vectors by using multiplex PCR. Out of 100 mosquito pools, 8 were positive for W. bancrofti (ERI of 0.33%) and 3 pools each were positive for D. immitis and D. repens (ERI 0.12%). The technique showed 100% sensitivity and 98% specificity. El-Nikhila, El-Matiaa villages, and Sahel Seleem district in Assiut Governorate, Egypt are still endemic foci for filarial parasites. Multiplex PCR offers a reliable procedure for molecular xenomonitoring of filariasis within their respective vectors in endemic areas. Therefore, it is recommended for evaluation of mosquito infection after lymphatic filariasis eradication programs.

Detection of Wuchereria bancrofti DNA in paired serum and urine samples using polymerase chain reaction-based systems

The Global Program for the Elimination of Lymphatic Filariasis (GPELF) aims to eliminate this disease by the year 2020. However, the development of more specific and sensitive tests is important for the success of the GPELF. The present study aimed to standardise polymerase chain reaction (PCR)-based systems for the diagnosis of filariasis in serum and urine. Twenty paired biological urine and serum samples from individuals already known to be positive for Wuchereria bancrofti were collected during the day. Conventional PCR and semi-nested PCR assays were optimised. The detection limit of the technique for purified W. bancrofti DNA extracted from adult worms was 10 fg for the internal systems (WbF/Wb2) and 0.1 fg by using semi-nested PCR. The specificity of the primers was confirmed experimentally by amplification of 1 ng of purified genomic DNA from other species of parasites. Evaluation of the paired urine and serum samples by the semi-nested PCR technique indicated only two of the 20 tested individuals were positive, whereas the simple internal PCR system (WbF/Wb2), which has highly promising performance, revealed that all the patients were positive using both samples. This study successfully demonstrated the possibility of using the PCR technique on urine for the diagnosis of W. bancrofti infection.

Evaluation of diagnostic tests for Wuchereria bancrofti infection in Brazilian schoolchildren

Introduction Since the launch of the Global Programme to Eliminate Lymphatic Filariasis, more than 70% of the endemic countries have implemented mass drug administration (MDA) to interrupt disease transmission. The monitoring of filarial infection in sentinel populations, particularly schoolchildren, is recommended to assess the impact of MDA. A key issue is choosing the appropriate tools for these initial assessments (to define the best intervention) and for monitoring transmission. Methods This study compared the pre-MDA performance of five diagnostic methods, namely, thick film test, Knott's technique, filtration, Og4C3-ELISA, and the AD12-ICT card test, in schoolchildren from Brazil. Venous and capillary blood samples were collected between 11 pm and 1 am. The microfilarial loads were analyzed with a negative binomial regression, and the prevalence and associated 95% confidence intervals were estimated for all methods. The accuracies of the AD12-ICT card and Og4C3-ELISA tests were assessed against the combination of parasitological test results. Results A total of 805 schoolchildren were examined. The overall and stratified prevalence by age group and gender detected by Og4C3-ELISA and AD12-ICT were markedly higher than the prevalence estimated by the parasitological methods. The sensitivity of the AD12-ICT card and Og4C3-ELISA tests was approximately 100%, and the positive likelihood ratios were above 6. The specificity of the Og4C3-ELISA was higher than that of the AD12-ICT at different prevalence levels. Conclusions The ICT card test should be the recommended tool for monitoring school-age populations living in areas with ongoing or completed MDA. .

Efeito do tratamento em massa com dietilcarbamazina na microfilaremia, antigenemia e anticorpos antifilariais em uma área endêmica do município de Olinda - PE / The effect of mass drug administration with single doses of diethylcarbamazine in microfilaremia, antigenemia e antifilarial antibodys in a endemic area in Olinda - PE

Este trabalho analisou o efeito do tratamento em massa com doses únicas anuais de Dietilcarbamazina (DEC), no período de 2007 a 2012, em indivíduos infectados por Wuchereria bancrofti, residentes em Olinda - PE. Para essa análise foram utilizadas as técnicas de filtração em membrana, na detecção da microfilaremia, o teste do cartão ICT e o Og4C3-ELISA, na detecção do antígeno circulante filarial, e o teste BM14 na avaliação dos níveis de anticorpos antifilariais. Os resultados obtidos indicam redução nas características avaliadas: após a quarta dose de DEC, a microfilaremia reduziu 100 por cento e a antigenemia pelo cartão ICT atingiu 78,1 por cento de redução após a quinta dose. A mediana do Og4C3 caiu significativamente de 7117 ua, para 1715 ua após a terceira dose, último ano que o teste foi realizado. Observou-se curva de redução também nos níveis de Bm14, com mediana da densidade ótica caindo de 2,1 para 0,1 após a quinta dose. A diminuição nas taxas das características estudadas indica que o tempo preconizado pela Organização Mundial de Saúde para a eliminação da transmissão da FL na área é suficiente para a negativação das microfilárias. Os resultados desse estudo mostram a elevada eficácia do esquema terapêutico utilizado no clareamento da microfilaremia e tratamento dos infectados, e sugerem que a utilização desse esquema na população possivelmente tenha levado a interrupção da transmissão na área. Sugere-se que haja um acompanhamento maior que cinco anos da população submetida ao tratamento para uma melhor avaliação dos níveis de anticorpos e de antigenemia filarial circulante...

Immunological characterization of recombinant Wuchereria bancrofti cuticular collagen (COL-4) as putative vaccine candidate for human lymphatic filariasis

OBJECTIVE@#To elucidate immunoprophylactic potential of recombinant Wuchereria bancrofti (W. bancrofti) cuticular collagen (COL-4) in BALB/c mice and filarial clinical samples.@*METHODS@#col-4 gene was PCR amplified from W. bancrofti L3 cDNA library and cloned in pRSET B vector. Recombinant COL-4 was over expressed in salt inducible system and was purified by nickel affinity chromatography. Humoral and cellular responses were measured by ELISA and peripheral blood mononuclear cells (PBMC) of various filarial clinical samples respectively using purified recombinant COL-4 antigen. Then the protective immune responses of COL-4 immunized BALB/c mice were characterized.@*RESULTS@#Sequence analysis of COL-4 with human host proteins reveals lack of homology. The recombinant COL-4 was found to be at 15 kDa fusion protein. The affinity purified COL-4 showed significant reactivity with putatively immune sera and in a similar fashion it demonstrated marked proliferation in PBMC samples. Immunization studies in experimental filarial host (mice) elicited significant titers with protective antibody isotype profile (IgM and IgG). Cellular immune responses were also significant in terms of splenocytes proliferation assay on mice samples.@*CONCLUSIONS@#Our immunological findings in experimental host suggest Th2 mediated immune response. Hence, we propose that W. bancrofti COL-4 could be an efficacious vaccine candidate against lymphatic filariasis.

Survey of Bancroftian filariasis infection in humans and Culex mosquitoes in the western Brazilian Amazon region: implications for transmission and control

Introduction The aim of this work was to identify possible lymphatic filariasis foci in the western Brazilian Amazonian that could be established from the reports of Rachou in the 1950s. The study was conducted in three cities of the western Brazilian Amazon region - Porto Velho and Guajará-Mirim (State of Rondônia) and Humaitá (State of Amazonas). Methods For human infection evaluation thick blood smear stained with Giemsa was used to analyze samples collected from 10pm to 1am. Polymerase chain reaction (PCR) was used to examine mosquito vectors for the presence of Wuchereria bancrofti DNA. Humans were randomly sampled from night schools students and from inhabitants in neighborhoods lacking sanitation. Mosquitoes were collected from residences only. Results A total 2,709 night students enrolled in the Program for Education of Young Adults (EJA), and 935 people registered in the residences near the schools were examined, being 641 from Porto Velho, 214 from Guajará-Mirim and 80 from Humaitá. No individual examined was positive for the presence of microfilariae in the blood stream. A total of 7,860 female Culex quinquefasciatus specimens examined were negative by PCR. Conclusions This survey including human and mosquito examinations indicates that the western Amazon region of Brazil is not a focus of Bancroftian filariasis infection or transmission. Therefore, there is no need to be included in the Brazilian lymphatic filariasis control program. .

Effects of midgut bacteria and two protease inhibitors on the reproductive potential and midgut enzymes of culex pipiens infected with wuchereria bancrofti

Laboratory investigations were carried out to asses the effect of some proteases inhibitors on the reproductive potential of Culex pipiens females resulted from larvae treated with different protease inhibitors. The fecundity and engorgement of symbiotic and aposymbiotic C. pipiens females were significantly reduced. The blood meal digestion period increased significantly. On the other hand, enzyme band with molecular weight of 40 KDa which may be cysteine protease was detected in untreated symbiotic and aposymbiotic female midguts. The results may explain that the absence of this enzyme bands in treated female midguts may be due to the inhibition caused by [E-64] a cysteine protease inhibitor

Effects of midgut bacteria and two protease inhibitors on the transmission of wuchereria bancrofti by the mosquito vector, culex pipiens

Laboratory investigations were carried out to study the effect of two protease inhibitors on the transmission of W. bancrofti filarial by Culex pipiens and to study the susceptibility interaction between filaria and protease inhibitors. The results obtained revealed that, infection rates were variable among untreated and treated symbiotic and aposymbiotic Cx. pipiens females resulted from third instar larvae treated with E-64 and EDTA. The survival rate was variable among untreated and treated symbiotic and aposymbiotic females resulted from third instar larvae treated with E-64 and EDTA. Protease inhibitor [E-64] caused inhibition of the parasite development and transmission by means of ceasing catalytic activity- responsible for parasite migration-caused by parasitic larval stages inside the mosquito vector

Evaluation of PCR-ELISA as a tool for monitoring transmission of Wuchereria bancrofti in District of Gampaha, Sri Lanka

<p><b>OBJECTIVE</b>To compare Wuchereria bancrofti (W. bancrofti) infection rates of Culex quinquefasciatus, using dissection and PCR-ELISA in two consecutive time periods (from 2007 to 2008 and from 2008 to 2009).</p><p><b>METHODS</b>Mosquitoes were collected in 30 sentinel and 15 non-sentinel sites in 15 Medical Officer of Health areas of Gampaha District known for the presence of W. bancrofti transmission in two consecutive time period of 2007 to 2008 and 2008 to 2009. Captured mosquitoes were dissected to determine the W. bancrofti larvae (L1, L2, L3). PCR was carried out using DNA extracted from mosquito pools (15 body parts/pool) utilizing the primers specific for Wb-SspI repeat. PCR products were analyzed by hybridization ELISA using fluorescein-labeled wild type specific probes. The prevalence of infected/infective mosquitoes in PCR pools (3 pools/site) was estimated using the PoolScreen™ algorithm and a novel probability-based method.</p><p><b>RESULTS</b>Of 45 batches of mosquitoes dissected, W. bancrofti infected mosquitoes were found in 19 and 13 batches, with an infection rate of 13.29% and 3.10% with mean larval density of 8.7 and 1.0 larvae per mosquito for two study periods in the Gampaha District. Total of 405 pools of head, thorax and abdomen were processed by PCR-ELISA for each year. Of these, 51 and 31 pools were positive for W. bancrofti in the two study periods respectively. The association of dissection based prevalence rates with PCR based rates as determined by the Pearson correlation coefficient were 0.176 and 0.890 respectively for the two periods.</p><p><b>CONCLUSIONS</b>Data indicate that PCR-ELISA is more sensitive than the traditional dissection techniques for monitoring transmission intensity.</p>

Rapid Detection and Identification of Wuchereria bancrofti, Brugia malayi, B. pahangi, and Dirofilaria immitis in Mosquito Vectors and Blood Samples by High Resolution Melting Real-Time PCR

A simple, rapid, and high-throughput method for detection and identification of Wuchereria bancrofti, Brugia malayi, Brugia pahangi, and Dirofilaria immitis in mosquito vectors and blood samples was developed using a real-time PCR combined with high-resolution melting (HRM) analysis. Amplicons of the 4 filarial species were generated from 5S rRNA and spliced leader sequences by the real-time PCR and their melting temperatures were determined by the HRM method. Melting of amplicons from W. bancrofti, B. malayi, D. immitis, and B. pahangi peaked at 81.5+/-0.2degrees C, 79.0+/-0.3degrees C, 76.8+/-0.1degrees C, and 79.9+/-0.1degrees C, respectively. This assay is relatively cheap since it does not require synthesis of hybridization probes. Its sensitivity and specificity were 100%. It is a rapid and technically simple approach, and an important tool for population surveys as well as molecular xenomonitoring of parasites in vectors.

Perfil epidemiológico e antigênico da população não aderente ao tratamento em massa para filariose linfática no município de Olinda - PE / Antigenic and epidemiological profile of the population adheres to mass treatment for lymphatic filariasis in the city of Olinda-PE

A Organização Mundial de Saúde aponta a Filariose Linfática como endemia potencialmente eliminável. Para isso, foi criado o Programa Global de Eliminação da Filariose Linfática, que possui como uma de suas estratégias o tratamento em massa da população endêmica. A baixa adesão ao tratamento representa um sério obstáculo ao sucesso do programa. Assim, o objetivo deste estudo é descrever o perfil epidemiológico e antigênico e os fatores relacionados a não adesão ao tratamento em massa para filariose linfática no município de Olinda-PE. Trata-se de um estudo de corte transversal, cuja população foi constituída por pessoas que se recusaram a participar do tratamento em massa nos bairros Alto da Bondade e Alto da Conquista, nos anos de 2009 e 2010. Os dados foram coletados através da aplicação de questionários e do teste antigênico (ICT card test). Os indivíduos positivos pelo ICT card test foram avaliados através da filtração (teste parasitológico padrão-ouro). Para o processamento e a análise de dados foi utilizado o programa Epi-Info. Foram pesquisados 102 indivíduos, sendo 51 (50,00 por cento) moradores do Alto da Bondade e 51 (50,00 por cento) do Alto da Conquista. A maioria era do sexo feminino (63,73 por cento) com média de idade de 49,39 anos, e possuía o ensino médio (17,6 por cento). Sobre as características socioambientais, verificou-se que todos os domicílios visitados eram de alvenaria. Cerca de 90por cento possuía abastecimento de água ligado à rede geral e coleta de lixo por serviço de limpeza urbana, porém o esgotamento sanitário era feito através de fossas rudimentares. Do total de pesquisados, quatro (3,92 por cento) apresentaram-se antígeno-positivos para Wuchereria bancrofti através do teste ICT card test), porém não apresentaram positividade na filtração. Os principais motivos encontrados para a não adesão ao tratamento em massa foram o não recebimento da medicação e relacionados às reações adversas, porém tais razões não foram completamente esclarecidas. Novas estratégias são necessárias para o aumento da adesão ao tratamento em massa, dentre as quais ações de educação em saúde e campanhas de sensibilização.