Characterization of serum cytokines and circulating microRNAs that are predicted to regulate inflammasome genes in cutaneous leishmaniasis patients.

Leishmaniasis is a neglected disease caused by an intracellular protozoan parasite of the genus Leishmania. Infection starts when this protozoan replicates in a phagolysosomal compartment in macrophages, after evading host immune responses. The balance of Th1 and Th2 immune responses is crucial in leishmaniasis because it will determine whether the infection will be under control or if clinical complications will occur. The inflammasome, which is activated during Leishmania infection, involves the action of caspase-1 and release of the proinflammatory cytokines interleukin-1ß and interleukin-18. Together, they contribute to the maintenance of an inflammatory response and pyroptosis. Here, we evaluated the serum levels of cytokines and the expression of circulating microRNAs related to inflammasome regulation in twenty-seven patients with cutaneous leishmaniasis in comparison to nine healthy individuals, in the context of the inflammasome activation. Evaluation of serum cytokines activation (IL-1ß, IL-2, IL-4, IL-6, IL-10, and IL-17) was performed by flow cytometry using CBA kits (cytometric beads array) while the expression of circulating microRNAs (miR-7, miR-133a, miR-146b, miR-155, miR-223, miR-328, and miR-342) in plasma was measured by quantitative polymerase chain reaction. Our results showed an increase of the expression of miR-7-5p (p < 10-5), miR-133a (p = 0.034), miR-146b (p = 0.003), miR-223-3p (p = 10-5), and miR-328-3p (p = 0.002), and cytokine levels for IL-1ß (p = 0.0005), IL-6 (p = 0.001), and IL-17 (p = 0.001) in patients with cutaneous leishmaniasis compared to the controls. These results suggest that microRNAs and cytokines can play an important role in regulating the human immune responses to Leishmania infection. Our findings may contribute to the understanding of the mechanisms of the gene regulation during the cutaneous leishmaniasis and to the identification of possible biomarkers of the infection.

Frequency and factors associated with Toxoplasma gondii infection in pregnant women and their pets in Ilhéus, Bahia, Brazil.

INTRODUCTION: Toxoplasmosis is an asymptomatic disease that can lead to systemic disease in the fetus of pregnant women with primary infection. This study aimed to determine the prevalence of toxoplasmosis, associated factors, and correlation between the serology of pregnant women and their pets, in the municipality of Ilhéus, Bahia, Brazil. METHODS: This cross-sectional study was conducted in 196 pregnant women and their cats or dogs (n=89). Semi-structured interviews were conducted and serum samples from the pregnant women were tested to detect IgM and IgG antibodies against Toxoplasma gondii, and avidity tests were performed for IgM-positive samples. The serum collected from pets were tested for IgG antibodies, and IgM antibodies in cats. A non-conditional logistic regression analysis was performed to identify infection-associated factors. RESULTS: IgG and IgM antibodies were detected in 67.9% (133/196) and 1.5% (3/196) samples, respectively, for women with an avidity of over 60%. Age ≥ 25 and the presence of cats in the vicinity were found to be associated with infection, while the level of education and previous orientation toward prevention of toxoplasmosis were protective factors in pregnant women. IgG antibodies were detected in 46.1% (41/89) of the animals, and cats were found to be negative for IgM. For the animals, age ≥ 1 year was a factor associated with infection. There was no correlation between serology of the pregnant women and the animals (p=0.15). CONCLUSIONS: An elevated prevalence of toxoplasmosis was detected in the region. Therefore, the adoption of preventive measures by public healthcare bodies is recommended.

Genetic structure of natural populations of the sand fly Lutzomyia longipalpis (Diptera: Psychodidae) from the Brazilian northeastern region.

In Latin America, Lutzomyia longipalpis is the principal vector of Leishmania chagasi, and is associated with the majority of active foci of visceral leishmaniasis. In spite of the fact that this sand fly is spread practically throughout the entire Neotropical Region, its distribution is not uniform due to geographic and environmental barriers. Geographic isolation coupled with reduced flight abilities may contribute to the appearance of cryptic species of Lutzomyia longipalpis, which may differ in their capacity to transmit L. chagasi. In this work, we describe the genetic structuring patterns based on polymorphism analysis of 24 RAPD-PCR loci of 7 natural populations of Lutzomyia longipalpis obtained from Brazil's northeastern region. The estimated degree of genetic differentiation between populations, based on the population subdivision index theta(ST) (0.136), suggests a moderate degree of genetic structuring as a result of geographical isolation and restricted gene flow. Genetic distances were found to be compatible with those found between members of a single species, suggesting a taxonomic uniformity of Lutzomyia longipalpis in the region studied.

[First report of Lutzomyia cortelezzii (Brèthes, 1923) in Ilhéus City, Bahia]. / Primeiro relato de Lutzomyia cortelezzii (Brèthes, 1923) na cidade de Ilhéus, BA.

Lutzomyia cortelezzii was found in the urban area of Ilhéus, Bahia, Brazil. The appearance this phlebotomine in 100% of 288 hours of capture in Ilhéus' territory is an unpublished fact. The collection was by CDC light traps between December 2003 and May 2004, with a total of 81 collected samples.

[Adaptation of the Rugai et al. method for analysis of soil parasites]. / Adaptação do método de Rugai e colaboradores para análise de parasitas do solo.

This study offers an adaptation of the Rugai method for the collection of cysts, eggs and parasite larvae from beaches, parks and public squares.

Frequency and factors associated with Toxoplasma gondii infection in pregnant women and their pets in Ilhéus, Bahia, Brazil

Abstract INTRODUCTION: Toxoplasmosis is an asymptomatic disease that can lead to systemic disease in the fetus of pregnant women with primary infection. This study aimed to determine the prevalence of toxoplasmosis, associated factors, and correlation between the serology of pregnant women and their pets, in the municipality of Ilhéus, Bahia, Brazil. METHODS: This cross-sectional study was conducted in 196 pregnant women and their cats or dogs (n=89). Semi-structured interviews were conducted and serum samples from the pregnant women were tested to detect IgM and IgG antibodies against Toxoplasma gondii, and avidity tests were performed for IgM-positive samples. The serum collected from pets were tested for IgG antibodies, and IgM antibodies in cats. A non-conditional logistic regression analysis was performed to identify infection-associated factors. RESULTS: IgG and IgM antibodies were detected in 67.9% (133/196) and 1.5% (3/196) samples, respectively, for women with an avidity of over 60%. Age ≥ 25 and the presence of cats in the vicinity were found to be associated with infection, while the level of education and previous orientation toward prevention of toxoplasmosis were protective factors in pregnant women. IgG antibodies were detected in 46.1% (41/89) of the animals, and cats were found to be negative for IgM. For the animals, age ≥ 1 year was a factor associated with infection. There was no correlation between serology of the pregnant women and the animals (p=0.15). CONCLUSIONS: An elevated prevalence of toxoplasmosis was detected in the region. Therefore, the adoption of preventive measures by public healthcare bodies is recommended.

Environmental parasitology: soil analysis for geohelminth larvae contamination in Ilhéus, Bahia

The study of parasitic soil contamination is important for monitoring biohazards in densely populated areas or those with high anthropic activities. The present study quantified geohelminth larvae in two estuaries in the urban area of Ilhéus, Bahia; one to the north, formed by the Almada River and another to the south, in the Pontal Bay. The collections were performed following standard methodology considering climactic conditions and local tide tables; the Rugai method with modifications was used to analyze soil sediments. Morphological results showed a significantly higher number of Strongyloides stercoralis larvae in the estuary of the Almada River from September to December, coinciding with higher temperatures and lower rainfall and humidity. However, there was significant variation in climatic conditions and in the classification of anthropic activity interfering in the frequency and diversity of soiltransmitted helminth larvae, which justifies its monitoring to ensure environmental health in areas frequented by residents and tourists in Ilhéus, Bahia.

[New outbreak of American tegumentary leishmaniasis in a military training center in the Zona da Mata region, in the north of the State of Pernambuco]. / Novo surto de leishmaniose tegumentar americana em área de treinamento militar na Zona da Mata norte do Estado de Pernambuco.

A new outbreak of American tegumentary leishmaniasis among military personnel is reported, with 71 cases confirmed by means of clinical, epidemiological and laboratory criteria. Seven samples were isolated and were identified as Leishmania (Viannia) braziliensis. The occurrence of outbreaks in this region confirms the endemic nature of this disease, and the magnitude of the occurrence seems to be related to non-adoption of individual protection measures.

Diversidade de flebotomíneos no Município de Ilhéus, Bahia / Diversity of phlebotomine Sand Flies in Ilheus, State of Bahia, Brazil

Objetivo: esse estudo visou identificar espécies de flebotomíneos em áreas rural e urbana de Ilhéus-BA, com registros humanos de leishmaniose tegumentar americana, e a possibilidade de infecção natural desses insetos e hamsters sentinelas. Metodologia: capturou-se com armadilhas luminosas tipo CDC e capturador tipo Castro, 4.476 flebotomíneos entre julho de 2007 e março de 2009, com 3.384 exemplares identificados morfologicamente, e os demais dissecados; hamsters foram expostos à picada dos insetos e avaliados quanto a susceptibilidade à infecção. Resultados: a identificação de espécies mostra a diversidade da fauna. Das 14 encontradas em área rural, as mais abundantes foram Lutzomyia fischeri (45,2 por cento), Lutzomyia whitmani (41,1 por cento) e Lutzomyia choti (9,84 por cento); na urbana, foram nove exemplares distribuídos entre quatro espécies. A dissecção das fêmeas não demonstrou infecção natural, e nenhum dos 42 hamsters revelou leishmanias em meio de cultivo. Conclusão: os dados indicam manutenção de ciclo zooótico de transmissão apenas em área rural.

Objective: this study aimed to identify species of phlebotomine in the rural and urban areas in Ilheus, State of Bahia, Brazil, cases of American Tegumentary Leishmaniasis in humans, and the possibility of natural infection of sentinel hamsters by those insects. Methodology: between July, 2007 and March, 2009, 4,476 phlebotomine were captured with CDC-type light traps and Castro aspirators. 3,384 were morphologically identified, and the others were dissected; hamsters were exposed to insects bite and evaluated according to susceptibility to infection. Results: the identification of species shows the diversity of the fauna. From the 14 species found in the rural area, the most prevalent were Lutzomyia fischeri (45.2percent), Lutzomyia whitmani (41.1percent) and Lutzomyia choti (9.84 percent); in the urban area nine specimens from four species were found. The dissection of females did not show natural infection and none of the 42 hamsters developed leishmania in culture environment. Conclusion: rural data associated with human notification show evidence of the maintenance of the zoonotic transmission cycle.

Novo surto de leishmaniose tegumentar americana em área de treinamento militar na Zona da Mata norte do Estado de Pernambuco / New outbreak of American tegumentary leishmaniasis in a military training center in the Zona da Mata region, in the north of the State of Pernambuco

Relata-se novo surto de LTA em militares com 71 casos confirmados pelos critérios clínico, epidemiológico e laboratorial. Obteve-se o isolamento de sete amostras, identificadas como Leishmania (Viannia) braziliensis. A ocorrência de surtos nesta região confirma o caráter endêmico, cuja magnitude parece estar relacionada a não adoção de medidas de proteção individual.

A new outbreak of American tegumentary leishmaniasis among military personnel is reported, with 71 cases confirmed by means of clinical, epidemiological and laboratory criteria. Seven samples were isolated and were identified as Leishmania (Viannia) braziliensis. The occurrence of outbreaks in this region confirms the endemic nature of this disease, and the magnitude of the occurrence seems to be related to non-adoption of individual protection measures.

Identificação molecular de Leishmania pela PCR em amostras do município de Ilhéus, Zona da Mata da Bahia, Brasil / Molecular identification of Leishmania by the PCR on samples from the municipality of Ilheus, Zona da Mata of Bahia, Brazil

A leishmaniose tegumentar é uma doença endêmica que apresenta distribuição em todo o Brasil. Na Bahia, verifica-se a ocorrência em todas as mesoregiões com maior incidência dos casos nos municípios de Ilhéus. Esse estudo teve como objetivo identificar Leishmania spp circulante na região. Foram coletadas 84 amostras de aspirado de borda de lesão de pacientes suspeitos de leishmaniose tegumentar, e inoculados em hamsters (Mesocricetus auratus). Os animais foram observados durante 3 meses e, após este período, foram eutanasiados e fragmentos do baço foram coletados para a visualização de amastigotas para o isolamento em cultura, extração de DNA e caracterização por reação em cadeia da polimerase (PCR). Duas amostras foram positivas para o alvo de 750 pb que corresponde ao minicírculo do kDNA, específico para o subgênero Viannia. Esses resultados e o fato de a Leishmania (Viannia) braziliensis ser uma espécie mais prevalente em área de Mata Atlântica indicam que a doença está associada a ess espécie do parasito no município de Ilhéus.

Caracterização da transmissão da Leishmaniose Tegumentar Americana no município de Ilhéus, Zona da Mata do Estado da Bahia / Characterization of transmission of American cutaneous leishmaniasis in Ilhéus, Zona da Mata of the State of Bahia

A Leishmaniose Tegumentar Americana é endêmica em todos os estados do Brasil. De acordo com os registros da Secretaria de Saúde municipal, Ilhéus, localizado no sul da Bahia, é endêmico para a doença. Esse estudo objetivou caracterizar a transmissão da enfermidade nesse município. Foi realizado um estudo retrospectivo entre os anos 2000 e 2006, proporcionando a realização de busca ativa de casos humanos em áreas rural e urbana (Santo Antonio e Teotônio Vilela), com um inquérito populacional em 147 e 77 indivíduos, respectivamente. Foram capturados e identificados 6.439 flebotomíneos com armadilhas CDC, mil deles separados em pools de 20 insetos e submetidos à Reação em Cadeia da Polimerase. Ademais, 1.528 fêmeas capturadas com armadilhas de Castro foram dissecadas. Foram atendidos 131 pacientes suspeitos da doença no Centro de Atenção Especializada III, entre Agosto/2007 e Setembro/2009, e avaliados clínica, epidemiológica, parasitológica, molecular e imunologicamente. Dos residentes na área urbana 79,4% visitaram, ao menos uma vez, a área rural. Foram identificadas 18 espécies de flebótomos, predominando Lutzomyia whitmani (52,49%) e L. fischeri (29,98%) na área rural, e L. cortelezzii (05 exemplares) na urbana. Não houve infecção natural à dissecção; e nenhum pool foi positivo à PCR. Mas, dos 84 hamsters inoculados com material humano, 03 amostras de esfregaços hepáticos apresentaram amastigotas; e 02 foram detectáveis por PCR para o subgênero Viannia. Não houve evidência de transmissão urbana, sugerindo uma origem rural dos casos, onde L. whitmani e L. fischeri possivelmente são vetores, tendo L. (V) braziliensis como agente etiológico nesse ciclo de transmissão. Recomenda-se intensificação na vigilância quanto à notificação de casos e atenção às características epidemiológicas locais.

Adaptação do método de Rugai e colaboradores para análise de parasitas do solo / Adaptation of the Rugai et al. method for analysis of soil parasites

Este trabalho propõe a utilização de uma adaptação do método de Rugai e colaboradores que se presta à coleta de cistos, ovos e larvas de parasitas das areias das praias, parques e praças públicas.

Primeiro relato de Lutzomyia cortelezzii (Brèthes, 1923) na cidade de Ilhéus, BA / First report of Lutzomyia cortelezzii (Brèthes, 1923) in Ilhéus City, Bahia

A Lutzomyia cortelezzii foi encontrada na área urbana da cidade de Ilhéus, Bahia, Brasil. A aparição deste flebotomíneo em 100 por cento das 288h de capturas no território ilheense é fato inédito. As coletas foram realizadas através de armadilhas luminosas tipo CDC, de dezembro/2003 a maio/2004, num total de 81 exemplares coletados.

Contaminação do solo por helmintos de importância médica na praia do sul (Milionários), Ilhéus - BA / Contamination of soil by helmints of medical importance in the south beach (Milionários) Ilhéus - BA

O estudo do nível de contaminação do solo por parasitos em áreas endêmicas, públicas e muito freqüentadas como as praias, faz-se necessário uma vez que o solo é uma fonte de infecção contínua, principalmente para as crianças. Para avaliar o nível de contaminação do solo por helmintos na praia do sul (Milionários), na cidade de Ilhéus - BA, utilizou-se o método de Rugai modificado aplicado a 24 amostras de solo, colhidas no mês de Maio em áreas amplamente movimentadas. Do total de larvas encontradas, 41,66% foram S. stercoralis e 58,83% foram de Ancilostomídeos, distribuídos em três distintas profundidades do solo: superfície, 10cm e 20cm. Esses resultados confirmaram a contaminação da praia por helmintos, concluindo-se que a mesma representa um importante foco de transmissão de infecções por larvas. Este estudo mostra a necessidade de implantação de um programa de controle dos parasitos mais freqüentes encontrados na área, bem como de um programa de conscientização e educação sanitária das comunidades.