Aflagellar Epimastigote of Trypanosoma caninum: Biological and Ultrastructural Study of this Atypical Evolutionary Form.

PURPOSE: Trypanosoma caninum exhibits atypical epimastigote forms under axenic conditions. This study aimed to analyze this evolutionary form under different cultivation conditions and provide more information about this evolutionary form. METHODS: We selected a T. caninum isolate with a high percentage of aflagellar epimastigote forms in axenic cultures. Two separate growth curves were generated for T. caninum cultured in Schneider axenic medium and co-cultured with the DH82 cell line, followed by analysis and quantification of evolutionary forms using bright field microscopy. In addition, ultrastructural analysis of T. caninum was performed under both cultivation conditions. RESULTS: The growth curves of T. caninum under axenic and co-cultivation conditions exhibited similar profiles. However, in the axenic culture, the number of parasites was three times higher at the peak of the exponential phase than in the co-culture. In contrast to that in the axenic culture, in which only the epimastigote forms were observed along the entire curve, during co-cultivation with the DH82 cell line, differentiation was observed for the trypomastigote and spheromastigote forms in low proportions. These results demonstrated that when cultured alone, the T. caninum isolate preserved the aflagellar epimastigote form, but in the presence of DH82 canine macrophages, they differentiated into evolutionary forms, particularly trypomastigote forms. Moreover, this study is the first to describe the presence of lipid bodies, structure described as the parasite's nutritional reserve, throughout the body of T. caninum. CONCLUSIONS: These findings describe biological and ultrastructural aspects of epimastigote aflagellar and suggest that this evolutionary form may be involved in the biological cycle of T. caninum, still unknown.

Insights into the proteomic profile and gene expression of Lutzomyia longipalpis-derived Lulo cell line.

BACKGROUND: Lutzomyia longipalpis-derived cell line (Lulo) has been suggested as a model for studies of interaction between sandflies and Leishmania. OBJECTIVES: Here, we present data of proteomic and gene expression analyses of Lulo cell related to interactions with Leishmania (Viannia) braziliensis. METHODS: Lulo cell protein extracts were analysed through a combination of two-dimensional gel electrophoresis and mass spectrometry and resulting spots were further investigated in silico to identify proteins using Mascot search and, afterwards, resulting sequences were applied for analysis with VectorBase. RESULTS: Sixty-four spots were identified showing similarities to other proteins registered in the databases and could be classified according to their biological function, such as ion-binding proteins (23%), proteases (14%), cytoskeletal proteins (11%) and interactive membrane proteins (9.5%). Effects of interaction with L. (V.) braziliensis with the expression of three genes (enolase, tubulin and vacuolar transport protein) were observed after an eight-hour timeframe and compared to culture without parasites, and demonstrated the impact of parasite interaction with the expression of the following genes: LLOJ000219 (1.69-fold), LLOJ000326 (1.43-fold) and LLOJ006663 (2.41-fold). CONCLUSIONS: This set of results adds relevant information regarding the usefulness of the Lulo cell line for studies with Leishmania parasites that indicate variations of protein expression.

Histopathological analysis of initial cellular response in TLR-2 deficient mice experimentally infected by Leishmania (L.) amazonensis.

Tegumentary leishmaniasis is an important public health problem in several countries. The capacity of the Leishmania species, at the initial moments of the infection, to invade and survive inside the host cells involves the interaction of surface molecules that are crucial in determining the evolution of the disease. Using C57BL/6 wild-type and TLR-2(-/-) mice infected with L. (L.) amazonensis, we demonstrated that TLR-2(-/-) mice presented eosinophilic granuloma in the ear dermis, different from C57BL/6 wild-type mice that presented a cellular profile characterized mainly by mononuclear cell infiltrates, besides neutrophils and eosinophils, during the two first week of infection. When the parasite load was evaluated, we found that the absence of TLR-2 lead to a significant reduction of the infection in deficient mice, when compared with C57BL/6 mice which were more susceptible to the infection. Using TLR-2 deficient mice, it was possible to show that the absence of this receptor determined the reduction of the parasite load and the recruitment of inflammatory cells during the two first weeks after L. (L.) amazonensis infection.

[Health education: teaching about American tegumentary leishmaniasis]. / Educação em saúde: ensinando sobre a leishmaniose tegumentar americana.

Knowledge on the circulation of American tegumentary leishmaniasis (ATL) and perception by the local population are highly relevant for developing a control program, since they mobilize the community for effective health measures. This work aimed to orient primary school teachers and pupils (grades 1 through 4) in the public school system in Maricá, Rio de Janeiro State, Brazil, on ATL, using talks, posters, and leaflets with an interactive approach. Talks were given in seven municipal schools, and activities were held with 1,314 pupils and 56 teachers. Leaflets were distributed to the pupils with activities to help consolidate the content. Two months later one school was chosen for post-evaluation. Of the 213 pupils who had attended the talk, 198 answered a questionnaire, of whom 178 (89.9%) could remember the disease and 125 (63.1%) provided correct answers on transmission. Thus, informed children and teachers can function as information disseminators in health within their community and are therefore capable of contributing to control of endemic diseases.

Educação em saúde: ensinando sobre a leishmaniose tegumentar americana / Health education: teaching about American tegumentary leishmaniasis

Os conhecimentos sobre a circulação da leishmaniose tegumentar americana (LTA), assim como sua percepção pela população local, são de grande valia para o estabelecimento de campanhas de controle, pois mobiliza a comunidade em ações sanitárias. Este trabalho teve como objetivo esclarecer alunos e professores do ensino fundamental (1ª a 4ª série) das Escolas Municipais de Maricá sobre a LTA, por meio de palestras, cartazes e cartilhas com atividades, visando à conscientização do grupo. Em sete Escolas Municipais, foram ministradas palestras e atividades a 1.314 alunos do ensino fundamental e 56 professores. Foram também distribuídas cartilhas aos alunos com atividades para a fixação do assunto. Após dois meses, uma escola foi eleita para avaliação (Escola E), sendo que, dos 213 alunos que assistiram a palestra, 198 responderam ao questionário, 178 (89,9 por cento) lembravam da doença, e 125 (63,1 por cento) responderam corretamente como ocorria a transmissão. Assim, crianças e professores informados podem funcionar como difusores da informação em Saúde na sua comunidade, sendo capazes de atuar de forma relevante no controle de endemias.