Imunocitoquímica e imunohistoquímica para o diagnóstico laboratorial da leishmaniose tegumentar / Immunocytochemistry and immunohistochemistry for laboratorial diagnosis of cutaneous leishmaniasis

0 aumento significativo do número de casos notificados da Leishmaniose Tegumentar Americana (LTA) e a expansão geográfica da endemia tem motivado o desenvolvimento de novas tecnologias para auxiliar no diagnóstico das leishmanioses, visando minimizar as restrições apresentadas pelos testes diagnósticos disponíveis nos serviços de saúde. 0 presente trabalho empregou imunocitoquimica e imunohistoquímica (ICQ/IHQ) como métodos diagnósticos laboratoriais para LTA. Amostras de culturas de Leishmania in vitro e cortes histológicos de lesões em animais infectados experimentalmente foram submetidos a ICQ/IHQ, utilizando anticorpos policlonais desenvolvidos para este estudo e o complexo avidina-biotina modificado (Ultra Streptavidin®). Em comparação com outras técnicas empregadas para o diagnóstico da LTA, nos casos avaliados, a IHQ apresentou resultados semelhantes aos da histopatologia com coloração HE, com sensibilidade de 33,3% para formas amastigotas. Quando considerada a presenva de antígenos de Leishmania no padrão celular, a IHQ apresentou uma sensibilidade de 83,3%, significativamente maior que na histopatologia e compatível com metodos padrão ouro de cultura e PCR. As metodologias de ICQ/IHQ desenvolvidas neste trabalho foram capazes de demonstrar em biópsias de lesões, a presença de formas amastigotas e antígenos de Leishmania, oferecendo contribuição adicional ao diagnóstico da LTA, sendo de facil aplicação e podendo ser utilizada no sistema público de saúde.

The significant increase in cutaneous leishmaniasis (CL) notified cases and the geographic expansion of this endemy has motivated the development of new techniques to help in leishmaniasis diagnosis, seeking the minimization of the restrictions imposed by the diagnostic tests available at the health services. The current study applied immunocytochemistry and immunohystochemistry methods (ICC/IHC) for laboratory diagnosis of CL. Imprints and histological sections from tissue infected with Leishmania were submitted to ICC/IHC methods using polyclonal antibodies developed for this study and a modified avidin-biotin complex (Ultra Streptavidin®). The samples also were submitted for routinely stained hematoxylin and eosin (H&E) specimens and gold standard methods (culture and PCR). Compared with other useful techniques for the CL diagnosis, ICC/IHC showed the same sensitivity results (33%) as H&E stain for amastigotes recognition. When the presence of Leishmania antigens was evaluated, ICC/IHC presented 83,3% sensitivity, i.e., higher than that detected by histopathology and equivalent with gold standard methods (culture and PCR). The ICC/IHC techniques developed in the current study were able to recognize amastigote forms and also Leishmania antigens in lesion biopsies, offering an additional help to CL diagnosis and it can be easily applied in the public health system.

Helminth eggs viability in sewage and biosolids sludge in Curitiba, Paraná, Brazil

Despite some benefits, municipal sludge poses hazards to human-kind and domestic animals because of the presence of some pathogenic agents in it. Bacteria, viruses, protozoan cysts, and helminth eggs may be present in the sludge. In Paraná State, SANEPAR, the responsible agency for collection and treatment of sewage sludge, is attempting to use the biosolids in agriculture. However, the risk to human and animal infection by the pathogenics should be considered before its use. An attempt was made to study the occurrence, viability and survival of helminth eggs and larva and protozoan cysts in the sludge. Parasitological analyses of twelve sludge biosolids and ten sewage sludge samples were collected and analyzed, from the treatment station ETE Belem (Curitiba, Brazil) for a period of one year. Four of the ten sewage sludge samples examined revealed the presence of helminth eggs as high as up to 20 eggs/l. In the biosolids sludge, the average number of helminth eggs, per 1 g of dry matter, was 4.85. However, the aerobic treatment reduced the viability of helminth egg to 56.67 per cent, and the total number of viable eggs was 1.85/g of dry matter. Ascaris sp. was the prevalent parasite (75.7 per cent), followed by H. diminuta (0.93 per cent), Trichuris sp. (7.4 per cent), Hymenolepis nana (3.6 per cent), Toxocara sp. (2.7 per cent), and Taenia sp. (1.3 per cent). Thus treatments will be necessary to hygienize the sludge for use in agriculture