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On the Indian subcontinent, visceral leishmaniasis (VL) is considered an anthroponosis. To determine possible reasons for its persistence during interepidemic periods, we mapped Leishmania infections among healthy persons and animals in an area of active VL transmission in Nepal. During 4 months (September 2007–February 2008), blood was collected from persons, goats, cows, and buffaloes in 1 village. Leishmania infections were determined by using PCR. We found infections among persons (6.1%), cows (5%), buffaloes (4%), and goats (16%). Data were georeferenced and entered into a geographic information system. The bivariate K-function results indicated spatial clustering of Leishmania spp.–positive persons and domestic animals. Classification tree analysis determined that among several possible risk factors for Leishmania infection among persons, proximity of Leishmania spp.–positive goats ranked first. Although our data do not necessarily mean that goats constitute a reservoir host of L. donovani, these observations indicate the need for further investigation of goats’ possible role in VL transmission.

Introducción. El análisis de la longitud de los fragmentos de restricción del producto amplificado y el estudio del ADN polimórfico amplificado al azar han demostrado ser herramientas útiles para la tipificación de Leishmania. Objetivos. Estudiar la utilidad de las técnicas moleculares para la identificación y tipificación de cepas de referencia de Leishmania spp. del Nuevo Mundo y valorar su aplicabilidad a muestras clínicas. Materiales y métodos. Se aplicó PCR para amplificar el gen que codifica la cisteíno-proteinasa B, y el análisis de la longitud de los fragmentos de restricción del producto amplificado utilizando ácido desoxirribonucleico de 16 cepas de referencia de Latinoamérica y de muestras clínicas de pacientes colombianos con leishmaniasis, y la técnica del ácido desoxirribonucleico polimórfico amplificado al azar utilizando ocho cepas de referencia. Se establecieron los patrones de bandas en cada caso. Resultados. Se obtuvo producto de amplificación en la PCR para Leishmania braziliensis, L. peruviana, L. panamensis y L. guyanensis. Para el resto, no fue posible amplificar el gen con los cebadores utilizados. La restricción mostró un patrón de bandas común para L. peruviana, L. guyanensis y L. panamensis, mientras L. braziliensis, presentaba un perfil individual único. El análisis de restricción del producto amplificado generó un patrón de bandas similar en los cinco pacientes estudiados, que se correspondía con el patrón generado por L. peruviana, L. guyanensis o L. panamensis. Mediante la amplificación al azar se obtuvieron patrones de bandas reproducibles con todas las cepas estudiadas, que posibilitaron la diferenciación. Se discuten las ventajas y limitaciones de ambos procederes. Conclusiones. El combinar ambas metodologías resultaría útil para identificar especies de importancia médica, tomando en cuenta sus ventajas y desventajas.

Neste artigo, discute-se o eixo metodológico utilizado na construção de modelo de vigilância de endemias em áreas urbanas, orientado por uma análise de situações de risco e por indicadores epidemiológicos espaciais. São apresentadas as premissas básicas do modelo, os critérios de seleção de variáveis sócio-econômicas e as etapas metodológicas necessárias na construção do indicador sintético de risco. Comenta-se, também, como algumas questões operacionais relativas à construção de mapas digitais de setores censitários e vinculação de bancos de dados foram equacionadas. Essa abordagem, incorporando o componente da organização do espaço na vigilância de doenças endêmicas, tendo como exemplo a hanseníase e a tuberculose, privilegia o uso integrado de sistemas de informação já existentes, na perspectiva de estratificar áreas urbanas diferenciadas que permite discriminar riscos desiguais para ocorrência de endemias. Essa é uma ferramenta para o planejamento e o gerenciamento das ações voltadas para o controle das endemias nas cidades.In this article we discuss the methodological issues associated with the creation of a surveillance system for endemic diseases in urban areas based on analysis of populations at risk and on spatially referenced epidemiological indicators. We comment on the system's basic requirements, selection criteria for socioeconomic variables, and methodological steps to combine these variables so as to construct a census-based deprivation index. We also present the ways we solved some operational problems related to generation of digitized census tracts maps and linkage of morbidity data from different sources. This approach, spatial organization into account in surveillance of endemic diseases, exemplified here by tuberculosis and leprosy, allows for the interaction of several official data sets from census and health services in order to geographically discriminate inner-city risk strata. Criteria for constructing these risk strata were considered a useful tool for health planning and management activities for the control of endemic diseases in cities.

BACKGROUND: Most of the Leishmania genome is reported to be constitutively expressed during the life cycle of the parasite, with a few regulated genes. Inter-species comparative transcriptomics evidenced a low number of species-specific differences related to differentially distributed genes or the differential regulation of conserved genes. It is of uppermost importance to ensure that the observed differences are indeed species-specific and not simply specific of the strains selected for representing the species. The relevance of this concern is illustrated by current study. METHODOLOGY/PRINCIPAL FINDINGS: We selected 5 clinical isolates of L. braziliensis characterized by their diversity of clinical and in vitro phenotypes. Real-time quantitative PCR was performed on promastigote and amastigote life stages to assess gene expression profiles at seven time points covering the whole life cycle. We tested 12 genes encoding proteins with roles in transport, thiol-based redox metabolism, cellular reduction, RNA poly(A)-tail metabolism, cytoskeleton function and ribosomal function. The general trend of expression profiles showed that regulation of gene expression essentially occurs around the stationary phase of promastigotes. However, the genes involved in this phenomenon appeared to vary significantly among the isolates considered. CONCLUSION/SIGNIFICANCE: Our results clearly illustrate the unique character of each isolate in terms of gene expression dynamics. Results obtained on an individual strain are not necessarily representative of a given species. Therefore, extreme care should be taken when comparing the profiles of different species and extrapolating functional differences between them.

Leishmania donovani is an intracellular protozoan parasite that causes visceral leishmaniasis (VL). Antimonials (SSG) have long been the first-line treatment against VL, but have now been replaced by miltefosine (MIL) in the Indian subcontinent due to the emergence of SSG-resistance. Our previous study hypothesised that SSG-resistant L. donovani might have increased in vivo survival skills which could affect the efficacy of other treatments such as MIL. The present study attempts to validate these hypotheses. Fourteen strains derived from Nepalese clinical isolates with documented SSG-susceptibility were infected in BALB/c mice to study their survival capacity in drug free conditions (non-treated mice) and in MIL-treated mice. SSG-resistant parasites caused a significant higher in vivo parasite load compared to SSG-sensitive parasites. However, this did not seem to affect the strains' response to MIL-treatment since parasites from both phenotypes responded equally well to in vivo MIL exposure. We conclude that there is a positive association between SSG-resistance and in vivo survival skills in our sample of L. donovani strains which could suggest a higher virulence of SSG-R strains compared to SSG-S strains. These greater in vivo survival skills of SSG-R parasites do not seem to directly affect their susceptibility to MIL. However, it cannot be excluded that repeated MIL exposure will elicit different adaptations in these SSG-R parasites with superior survival skills compared to the SSG-S parasites. Our results therefore highlight the need to closely monitor drug efficacy in the field, especially in the context of the Kala-azar elimination programme ongoing in the Indian subcontinent.

BACKGROUND: Leishmania donovani is an intracellular protozoan parasite that causes a lethal systemic disease, visceral leishmaniasis (VL), and is transmitted between mammalian hosts by phlebotomine sandflies. Leishmania expertly survives in these 'hostile' environments with a unique redox system protecting against oxidative damage, and host manipulation skills suppressing oxidative outbursts of the mammalian host. Treating patients imposes an additional stress on the parasite and sodium stibogluconate (SSG) was used for over 70 years in the Indian subcontinent. METHODOLOGY/PRINCIPAL FINDINGS: We evaluated whether the survival capacity of clinical L. donovani isolates varies significantly at different stages of their life cycle by comparing proliferation, oxidative stress tolerance and infection capacity of 3 Nepalese L. donovani strains in several in vitro and in vivo models. In general, the two strains that were resistant to SSG, a stress encountered in patients, attained stationary phase at a higher parasite density, contained a higher amount of metacyclic parasites and had a greater capacity to cause in vivo infection in mice compared to the SSG-sensitive strain. CONCLUSIONS/SIGNIFICANCE: The 2 SSG-resistant strains had superior survival skills as promastigotes and as amastigotes compared to the SSG-sensitive strain. These results could indicate that Leishmania parasites adapting successfully to antimonial drug pressure acquire an overall increased fitness, which stands in contrast to what is found for other organisms, where drug resistance is usually linked to a fitness cost. Further validation experiments are under way to verify this hypothesis.

Neste artigo, discute-se o eixo metodológico utilizado na construção de modelo de vigilância de endemias em áreas urbanas, orientado por uma análise de situações de risco e por indicadores epidemiológicos espaciais. São apresentadas as premissas básicas do modelo, os critérios de seleção de variáveis sócio-econômicas e as etapas metodológicas necessárias na construção do indicador sintético de risco. Comenta-se, também, como algumas questões operacionais relativas à construção de mapas digitais de setores censitários e vinculação de bancos de dados foram equacionadas. Essa abordagem, incorporando o componente da organização do espaço na vigilância de doenças endêmicas, tendo como exemplo a hanseníase e a tuberculose, privilegia o uso integrado de sistemas de informação já existentes, na perspectiva de estratificar áreas urbanas diferenciadas que permite discriminar riscos desiguais para ocorrência de endemias. Essa é uma ferramenta para o planejamento e o gerenciamento das ações voltadas para o controle das endemias nas cidades.

Neste artigo, discute-se o eixo metodológico utilizado na construção de modelo de vigilância de endemias em áreas urbanas, orientado por uma análise de situações de risco e por indicadores epidemiológicos espaciais. São apresentadas as premissas básicas do modelo, os critérios de seleção de variáveis sócio-econômicas e as etapas metodológicas necessárias na construção do indicador sintético de risco. Comenta-se, também, como algumas questões operacionais relativas à construção de mapas digitais de setores censitários e vinculação de bancos de dados foram equacionadas. Essa abordagem, incorporando o componente da organização do espaço na vigilância de doenças endêmicas, tendo como exemplo a hanseníase e a tuberculose, privilegia o uso integrado de sistemas de informação já existentes, na perspectiva de estratificar áreas urbanas diferenciadas que permite discriminar riscos desiguais para ocorrência de endemias. Essa é uma ferramenta para o planejamento e o gerenciamento das ações voltadas para o controle das endemias nas cidades.