Luis Eduardo Ramirez

High prevalence of Trypanosoma rangeli and Trypanosoma cruzi in opossums and triatomids in a formerly-endemic area of Chagas disease in Southeast Brazil

In Brazil Trypanosoma rangeli has been detected in humans, sylvatic mammals and vectors in the Amazon Basin and in wild rodents in a Southern State. Here we report for the first time a high prevalence of T. rangeli in opossums and triatomids captured in peridomestic environments in a formerly-endemic area of Chagas disease in Southeast Brazil. Five molecular typing tools clearly indicate the presence of T. rangeli and Trypanosoma cruzi in mammalian reservoirs and triatomids. Twenty-one opossums (Didelphis albiventris) were captured and flagellates were detected in the blood of 57.1% (12/21) of the animals. Single infections with T. rangeli or T. cruzi were diagnosed, respectively, in 58.4 and 8.3% of the opossums. Mixed infections were observed in 33.3%. Forty-four triatomids (38 Rhodnius neglectus and 6 Panstrongylus megistus) were collected in palm trees within 50 m from human dwellings. Flagellates were observed in the digestive tract and feces of 50% of the insects. PCR assays performed in DNA samples obtained from 16 cultures of the intestinal tract revealed single infection with T. cruzi (68.7%) or T. rangeli (6.3%), as well as mixed infections (25%). T. rangeli was also detected in the hemolymph of two specimens. Genotyping revealed predominance of T. cruzi I. The data suggest that R. neglectus in conjunction with D. albiventris may be significant factors in the maintenance of the sylvatic and peridomestic cycles of T. rangeli in the region. The finding of T. cruzi and T. rangeli in triatomine species capable of domiciliation and therefore considered as alternative vectors for the parasite transmission opens up the possibility of re-establishment of Chagas disease following reinfestation of houses.

The hamster (mesocricetus auratus) as experimental model in chagas' disease: parasitological and histopathological studies in acute and chronic phases of Trypanosoma cruzi infection

This research characterizes the acute and chronic phases of Chagas disease in hamster through parasitological and histopathological studies. The acute phase was achieved with 44 young hamsters injected intraperitoneally with 100,000 blood trypomastigotes of Benedito and Y strains of T. cruzi. The chronic phase was induced in 46 hamsters injected intraperitoneally with 35,000 trypomastigotes of Vicentia, Benedito and Y strains. Animals were sacrificed at regular intervals of 24 hours of acute phase and from the 3rd to the 10th month of infection of chronic phase. In the acute phase, parasites were easily recovered from all animals and there was an inflammatory reaction characterized by mononuclear and polymorphous leukocyte infiltration of variable degree in the majority of tissues and organs, specially in the connective loose and fatty tissues, smooth muscle myocardium and skeletal muscle. In the chronic phase the lesions occurred in the same tissues and organs, but the inflammatory response was less severe and characterized by mononuclear infiltration mainly with focal or zonal fibrosis in the myocardium. In 50% of infected animals parasites were found in myocardium and recovered from pericardic, peritoneal and ascitic fluids in some animals. Signs of heart failure, sudden death and enlargement of bowel were observed regularly. We concluded that the hamster is an useful model for Chagas disease studies.

Primeira evidência de Trypanosoma rangeli no sudeste do Brasil, região endêmica para doença de Chagas

This short communication informs the discovery of Trypanosoma rangeli for the first time at Triângulo Mineiro region, South-east of Brazil, a highly endemic area of Chagasdisease and also the natural infection of Didelphis albiventris with the same trypanosome. Both the findings were demonstrated through blood smears, xenodiagnosis, microhematocrit technics and PCR. The last one was realized in faeces and hemolymph of Triatoma infestans utilizing as controls strains of T. rangeli from Colombia.

Effect of protective and non‐protective antibodies in the phagocytosis rate of Trypanosoma cruzi blood forms by mouse peritoneal macrophages

Summary The phagocytosis of Trypanosoma cruzi blood forms by mouse peritoneal macrophages is significantly enhanced by sera from chronic chagasic patients, rabbits and mice presenting ‘lytic antibodies’ (LA) which are associated with resistance and active infections as well as ‘conventional serology antibodies’ (CSA) which are immunoglobulins involved in the positivity of serological diagnostic tests. The phagocytosis rate, however, is not influenced by sera from mice immunized with T. cruzi antigen or chagasic patients submitted to specific treatment, both displaying only CSA but not LA. The efficacy of LA in increasing phagocytosis is related to their ability to bind to epitopes of living trypomastigotes, a property lacking in CSA that bind only to fixed parasites. This phenomenon is apparently the reason for the low effectiveness of antigens used for vaccination in Chagas disease which only induce CSA, immunoglobulins apparently unable to mediate a number of regular effector immune mechanisms such as complement‐mediated lysis, antibody‐dependent cell cytotoxicity and phagocytosis.

Cardiac neuronal depopulation in hamsters (Mesocricetus auratus) chronically infected with Trypanosoma cruzi

The aim of this study was to obtain an experimental animal model of destruction of cardiac neurons in order to investigate the behavior of the cardiac nervous system of hamsters chronically infected with Trypanosoma cruzi. We counted the neuronal cells of the cardiac autonomic nervous plexus in hamsters inoculated with 35,000 blood forms of three different T. cruzi strains and killed 5, 8 and 10 months after infection. We showed for the first time severe neuronal destruction in an experimental animal model with characteristics similar to those observed in human Chagasdisease.

Pancreatic hepatocytes in hamsters (Mesocricetus auratus) infected with Trypanosoma cruzi

Hepatocytic metaplasia may be induced in hamsters by carcinogens, and associated with aging, diabetes or chronic pancreatitis. By means of histopathologic and immunohistochemic studies, we observed pancreatic hepatocytes in hamsters infected and reinfected with Trypanosoma cruzi. The change was seen in 18 (19%) out of 94 infected animals, and was not found among 53 controls, Normal islet cells were immunoreactive for neuron-specific enolase and not reactive for NCL-HAS. Metaplastic cells were immunoreactive for NCL-HAS and not reactive for islet hormones and enolase. No relationship was observed between number of inoculations and metaplasia; however, the intensity of the inflammatory process and sequels seems to favor the development of metaplastic cells. Hamsters infected with T. cruzi may be useful to study hepatocytic metaplasia, and contribute to clarify aspects of Chagas disease and pancreatic changes. Our data indicate that aging, in addition to inflammation and atrophy, plays a role in this change.

Hepatocyte metaplasia in experimental chagasic pancreatitis: preliminary report

Beginning the study of chronic pathologic changes in pancreas of hamsters experimentally infected with Trypanosoma cruzi Vic strain, hepatocyte metaplasia was observed in one animal from infected group. This is the first report of oncocytes in Chagas disease, which could be due to aberrant regenerative response to pancreas inflammatory process.

Avaliação de uma nova microtécnica de fixação de complemento (Garcia & Sotelo, 1991) para o diagnóstico da doença de Chagas crônica com diferentes preparações antigênicas

From this present data it has been evaluated a new complement fixation test, comparatively to indirect immunofluorescence to diagnose chronic Chagas disease, utilizing one watery extract of epimastigotes of Trypanosoma cruzi and three other ethanolic extracts: one from epimastigotes, one from tripomastigotes and a third one of amastigotes obtained from cultures. Utilizing 236 serum samples indirect immunofluorescence test was performed: 109 positives (20 of them with positive parasitologic diagnostic) and 127 negatives (96 of healthy blood donors and 31 with other diseases). The results have showed that is possible a positive reaction in diluted samples up to 1:16. The best limits of reactivity found were the dilutions 1:4 for the ethanolic extract of amastigotes and 1:2 for the others antigens. The correlation index among the new complement fixation test and indirect immunofluorescence test showed that the ethanolic extract from epimastigotes was the best antigen to be utilized to diagnosis purposes. Its co-positivity index with indirect immunofluorescence was 0.92207 and the co-negative index was 0.90000. Concluding, the new complement fixation test showed itself as a fast, sensible, easily applicable semiquantitative microtechnique to the diagnosis of chronic Chagas disease.