PCR-based diagnosis for detection of Leishmania in skin and blood of rodents from an endemic area of cutaneous and visceral leishmaniasis in Brazil.

The technique of polymerase chain reaction (PCR) associated to hybridization was used to screen 123 samples collected from wild and synanthropic rodents captured in a cutaneous and visceral leishmaniasis endemic area in the State of Minas Gerais, Brazil. The detection of Leishmania spp in naturally infected rodents is of fundamental importance for incriminating them as possible reservoir hosts of the diseases in Minas Gerais. A total of 62 specimens belonging to wild (Thrichomys apereoides, Oryzomys subflavus, Galea spixii, Bolomys lasiurus and Wiedomys pyrrhorhinos) and synanthropic (R. rattus) rodent species were captured in different ecotopes. Blood and skin samples were submitted for PCR analyses followed by molecular hybridization with specific probes for the three Leishmania-species complexes. Fifteen samples were found positive after PCR-hybridization and identified as follows: nine belonging to the L. mexicana complex, three to the L. braziliensis complex and three to the L. donovani complex. Positive PCR results were found in 11 out of the 61 (18%) blood samples and in four out of the 62 (6.4%) skin fragments screened. R. rattus and T. apereoides were the most abundant species in the area also presenting high prevalence of natural infection. The presence of parasite DNA belonging to L. braziliensis, L. mexicana and L. donovani complexes was confirmed in several individuals of a rodent species, R. rattus. This work is the first report of the detection of L. (L.) chagasi in a naturally infected T. apereoides. The utility of filter paper as a substrate for PCR analyses and the efficacy of the procedure associated to the hybridization is emphasized.

Chagas disease and HIV co-infection: genetic analyses of two Trypanosoma cruzi strains under experimental immunosuppression.

BACKGROUND: Recently new aspects of the immunopathology of Chagas disease have been described in patients infected with HIV and unusual clinical manifestations such as cutaneous lesions, involvement of central nervous system and/or serious cardiac lesions related to the reactivation of the parasite have been reported. Two uncloned Trypanosoma cruzi strains previously isolated from chronic chagasic patients with HIV co-infection were studied in order to evaluate the impact of the immunosuppression on the genetic diversity of the parasite. RESULTS: We have exploited an experimental model to determine whether genetically distinct populations appear after immunosuppression as a consequence of in vivo selection or in vitro propagation. The in vitro and in vivo conditions have allowed us to study the selected populations. The first strain was isolated from a case of reactivation of Chagas disease in a patient which presented four cerebral lesions. It was possible to demonstrate that the patient was infected with at least three distinct populations of T. cruzi. The population, recovered after immunosuppression, in mice was genetically divergent from the primary human isolate. The second strain, isolated from a hemophiliac/HIV positive patient presenting cardiac manifestation of Chagas disease showed no marked genetic difference after experimental immunosuppression. CONCLUSION: The immunological condition of the patient, associated or not to the reactivation of the infection, and also the strain of the parasite may have an important role during the course of the disease. The in vivo mechanism that generates parasite genetic variability or the participation of the selection under stress conditions will require further investigation.

Genetic heterogeneity in Trypanosoma cruzi strains from naturally infected triatomine vectors in northeastern Brazil: epidemiological implications.

Eighteen Trypanosoma cruzi strains isolated from naturally infected triatomines were studied genetically. The majority of the strains were from Triatoma brasiliensis, the principal vector of Chagas disease in the northeast of Brazil. Multilocus enzyme electrophoresis (MLEE) and randomly amplified polymorphic DNA (RAPD) analyses were used to investigate the genotypic diversity and the spread of the T. cruzi genotypes in different environments. MLEE clearly distinguished two distinct isoenzyme profiles, and RAPD analysis revealed 10 different genotypes circulating in rural areas. The strains could be typed as isoenzyme variants of the T. cruzi principal zymodeme Z1 (T. cruzi I). An effective program of epidemiological vigilance is required to prevent the spread of T. cruzi I strains into human dwellings.

Chagas'disease and HIV co-infection: genotypic characterization of the Trypanosoma cruzi strain

In the past few years, new aspects of the immunopathology of Chagas' disease have been described in immunosuppressed patients, such as fatal central system lesions related to the reactivation of the parasite. This article is the first description of the genotypic characterization, at the strain level, of Trypanosoma cruzi isolated from a patient with Chagas' disease/AIDS co-infection. The presence of four hypodense lesions was observed in the cranial compute tomographic scan. The diagnosis of AIDS was assessed by the detection of anti-HIV antibodies using enzyme-linked immunosorbent assay (ELISA) and Western blot techniques. The CD4+ lymphocyte counts were maintained under 200 cells/mm cube number during one year demonstrating the severity of the state of immunosuppression. Chagas' disease was confirmed by serological and parasitological methods. Trypomastigote forms were visualized in a thick blood smear. The parasite isolated is genotypically similar to the CL strain. The paper reinforces that cerebral Chagas' disease can be considered as another potencial opportunistic infection in AIDS resulting from the reactivation of a dormant T. cruzi infection acquired years earlier.

Casos de leishmaniose tegumentar americana por Leishmania (Viannia) braziliensis nos Municípios de Cosmópolis e Indaiatuba-regiäo de Campinas, Estado de Säo Paulo, Brasil / Cases of mucocutaneous leishmaniasis due to Leishmania (Viannia) braziliensis from Cosmópolis and Indaituba municipalities - Campinas region, State of Säo Paulo, Brazil

A study was carried out to identify Leishmania species involved in skin lesions of patients from Cosmópolis and Indaiatuba, State of Säo Paulo, Brazil. The epidemiological data of cutaneous leishmaniasis in two cities suggested a epidemic situation in 1994. The lesions were clinically characteristic of cutaneous leishmaniasis and five out six patients responded positively to Montenegro's intradermal test. The histopathology of skin lesions were characterized by two patterns: exudative-cellular reaction and exudative granulomatous reaction. The clinical and histopathological parameters suggested Leishmania (Viannia) braziliensis as the possible etiologic agent. In agreement, it was difficult to isolate and maintain the parasite in the laboratory. Characterization by in situ hybridization with kDNA amastigotes from lesions fragments confirmed that Leishmania (Viannia) braziliensis was the parasite responsible for the studied cutaneous lesions.