Chronic myocardial damage in experimental T. cruzi infection of a new world primate, Cebus sp. monkey

Eighteen Cebus apella monkeys, (juvenile and adult of both sexes) were inoculated five years ago, with three Trypanosoma cruzi strains (CA1, n = 10; Colombian, n=4 and Tulahuen, n=4), either by conjunctival or intraperitoneal route, once or repeatedly. Parasitological, hematological, serological, enzymatic, radiographic, electro and echocardiographic findings have been peviously published15 and they are similar to those observed in human pathology. The most frequent electrocardiographic alteration was right branch bundle block. Six animals, chosen at random, were sacrificed. Those sacrificed 20 to 25 months post-first inoculation showed focal accumuli of leukocytes with myocytolysis. Foci of diffuse interstitial fibrosis with mild infiltrate of leukocytes among fibers were observed in the animals sacrificed 36 to 47 months post-inoculation. No parasites were seen. The lesions were more prominent in the ventricular walls and the septum. The fact that the infiltrates were predominant in the animals sacrificed at a shorter time after first inoculation and that fibrosis was more severe in those sacrificed at a longer time suggests that there is a progression of the infiltrative lesions to fibrosis, with a leukocytic activity indicative of a chronic phase. These lesions are similar to those described in human chronic Chagas' disease. This would demonstrate that this model is useful in evaluating a progress in the knowledge of the pathogenesis which is still a controversial issue, immunology, immunogenesis and chemotherapeutic agents of the chronic and indeterminate phases of this disease.

Efecto de la reinfección sobre la evolución de ratas infectadas con Trypanosoma cruzi

El objetivo de este trabajo fue comprobar si una de las variables medio-ambientales, la reinfección, puede modificar el comportamiento observado en un modelo de rata a nivel de parasitemia, anticuerpos séricos, manifestaciones electrocardiográficas y/o lesión miocárdica. Los grupos experimentales fueron: GI: ratas infectadas al destete con 1 x 10(6) T. cruzi; GR: igual a GI más reinfecciones cada 30 días hasta los 150 días post-infección inicial (p.i.i.); GI1 ratas de 51 días infectadas; GT: testigos. Se detectó parasitemia alta en GI y GR hasta los 20 días p.i.i. tendiendo a negativizarse al día 30. En GR no se observaron parásitos despúes del primer reinóculo, resistencia que no es debida sólo a la mayor edad del huésped pués hubo parasitemia en GI1. Los xenodiagnósticos fueron negativos en los tres grupos. Los anticuerpos séricos no se modificaron significativamente en GR respecto de GI, salvo en los anticuerpos 7S, pues los del GR presentaron títulos superiores en algunos de los días estudiados. Los ECG basales no mostraron cambios distintivos en las ratas infectadas. La prueba de ajmalina mostró una disminución de la FC independiente del tratamiento; el PR, QaT y QRS se prolongaron significativamente en todos los grupos respecto del basal (p < 0.05), salvo el QaT en el GT; además, el cambio de PR y QaT fue mayor en los infectados (p < 0.05). En los grupos infectados hubo también una amplia variación en la orientación del eje eléctrico respecto del valor basal, acompañado de cambios morfológicos más manifiestos en GR. La proporción de lesión cardíaca detectada histológicamente en los grupos infectados, fue significativamente superior a la del GT (p < 0.01). Se concluye que la reinfección no reproduce el cuadro agudo inicial y no modifica el tipo y grado de lesión cardíaca observada histológicamete. La prueba de ajmalina muestra cambios electrocardiográficos compatibles con daño miocárdico incipiente en las ratas infectadas y sugieren mayor compromiso en el grupo reinfectado.

Observações sobre a viabilidade do Trypanosoma cruzi no caldo da cana

Em Catolé do Rocha (Estado da Paraíba, Brasil) várias pessoas foram concomitantemente acometidas de doença de Chagas, em fase aguda. A propósito desse evento, houve cogitação de que o Trypanosoma cruzi foi veiculado pela via digestiva, através de ingestão de caldo de cana acidentalmente contaminado. Com o presente estudo, verificamos que o parasita permaneceu viável, durante quatro horas, no líquido em questão mantido em temperatura ambiente. Ao mesmo tempo, comprovamos que após uma hora ele pôde promover infecção em camundongo ao ser administrado por meio de sonda gástrica, renovando o interesse já demonstrado quanto a esse mecanismo alternativo de transmissão.

Simultaneous identification of Trypanosoma cruzi surface and internal antigens reactive to different immunoglobulin classes (radio-immunoblotting)

A radioactive Western-blotting technique was developed by which the reactivity of Immunoglobulins (Igs) from different classes to both membrane radiolabelled and internal parasite antigens is simultaneously identified. The method includes radioiodination of parasites, polypeptide fractionation by SDS-PAGE, Western-blot transfer and autoradiography of the immunoblots developed with anti-Igs conjugates labelled with enzymes. The analysis is then performed by the comparison of common bands on the autoradiograms and the respective substrate stained nitrocellulose blots. This technique was used to analyse T. cruzi trypomastigote surface labelled antigens reactive to IgM, IgA and IgG specific antibodies. A different pattern of reactivity with acute Chagas' disease patients sera was thus obtained.

Trypanosoma cruzi: modification of alkaline phosphatase activity induced by trypomastigotes in cultured human placental villi

Human term placental villi cultured ''in vitro" were maintained with bloodstream forms of Trypanosoma cruzi during various periods of time. Two different concentrations of the parasite were employed. Controls contained no T. cruzi. The alkaline phosphatase activity was determined in placental villi by electron microscopy and its specific activity in the culture medium by biochemical methods. Results showed that the hemoflagellate produces a significant decrease in enzyme activity as shown by both ultracytochemical and specific activity studies and this activity was lower in cultures with high doses of parasites. The above results indicate that the reduction in enzyme activity coincides with the time of penetration and proliferation of T. cruzi in mammalian cells. These changes may represent an interaction between human trophoblast and T. cruzi.

Epidemiological survey of Trypanosoma cruzi infection in North-Eastern Brazil using different diagnostic methods

A survey of the prevalence of Trypanosoma cruzi infection was carried out in Oitis, a small community in the State of Piaui, Brazil. Two hundred and sixty five individuals were screened by microscopic examination, hémoculture, indirect immunofluorescence (IFA), enzyme-linked immunosorbent assay (ELISA), and competitive enzyme-linked immunosorbent assay (C-ELISA) using the monoclonal antibody TCF87 against to a 25kd T. cruzi antigen. Seropositivity was 14.3% by the IFA test, 14.7% by ELISA, and 13.2% by C-ELISA. The C-ELISA using the TCF87 monoclonal antibody seems to be applicable in serodiagnosis of Chagas' disease.

Novo processo para triagem de medicamentos na infecção experimental pelo Trypanosoma cruzi

É proposto processo para a triagem da capacidade terapêutica de medicamentos na infecção experimental pelo Trypanosoma cruzi. O método tem base no emprego de triatomíneos parasitados que se alimentam, decorridos períodos diferentes para haver compatibilização com níveis sangüíneos, em camundongos aos quais foi administrado o fármaco sob apreciação; assim, o tubo digestivo do hemíptero participará como estrutura propícia à avaliação. Em observação inicial, ocorreu utilização do benzonidazol, que se mostrou apenas parcialmente ativo, pelo menos de acordo com a maneira de execução do novo procedimento.

Evolution of subpatent parasitaemia in Trypanosoma cruzi chronically infected mice with the help of a cyclophosphamide amplification transfer assay

We have evaluated the sensitivity of the classical blood subinoculation method, modified through cyclophosphamide treatment of transferred mice, for the detection of occult parasitaemias in Trypanosoma cruzi chronically infected mice. Besides its simplicity, the method was shown to be highly sensitive for both the "chronic" phase parasites (99% of chronic cases were shown to harbour occult parasitaemias) and for the acute phase parasites (T. cruzi could be detected in 53.8% of animals transferred with one Y strain parasite and in 20% of animals transferred with one CL strain parasite). Using acute phase bloodforms, the assay proved to be more sensitive than conventional subinoculation when dealing with the CL, but not the Y strain of the parasite. With the help of this parasite detection tool, we have studied during a one year period, the evolution of subpatent parasitaemias in a group of mice which survived through chemotherapy from lethal acute phase of T. cruzi infection. Cyclophosphamide transfer assay revealed occult parasitaemias in 100% of the chronic animals, nevertheless, continuous and discontinuous patterns of positivity were observed.

Evaluación del método de ELISA para detección de antígenos y complejos inmunes circulantes de Trypanosoma cruzi a través de un estudio de campo en una zona endémica de Argentina

En una zona endémica de la República Argentina se llevó a cabo un ensayo de campo de la prueba inmunoenzimática ELISA para la detección de antígenos (cAg) y complejos inmunes circulantes (CIC) en sueros de pacientes chagásicos crónicos. Del total de 215 muestras de sangre analizadas, 51 fueron positivas para ELISA-CIC y 45 lo fueron para ELISA-cAg. De los 74 (34,32% de la población) sujetos considerados infectados con dos reacciones serológicas positivas, 49 (66,21%) presentaron CIC en suero, en tanto que en 43 (58,11%) de ellos se encontró cAg por ELISA. Solo en 2 casos serológicamente no reactivos, se detectaron inespecíficamente CIC y cAg. Dentro del grupo considerado no infectado, se observó reactividad inespecífica de bajo título por una de las pruebas serológicas en 16 (11,35%) de 141 individuos. Estos sueros arrojaron resultados consistentemente negativos por ELISA-CIC y cAg demostrando la utilidad de estos métodos de diagnóstico antigénico en casos de serología conflictiva. La determinación de fracciones antigénicas circulantes por ELISA en individuos chagásicos crónicos permite evidenciar la infección por T. cruzi de manera más directa que midiendo la respuesta inmune humoral en el huésped, presentando además mayor sensibilidad que el diagnóstico parasitológico clásico

Avaliação da atividade terapêutica do itraconazol nas infecções crônicas, experimental e humana, pelo Trypanosoma cruzi

Alguns pesquisadores demonstraram que o intraconzol, "in vitro" e na infecção aguda de animais, possui atividade antiparasitária referente ao Trypanosoma cruzi. Diante dessas observações, decidimos empreender estudo sobre e etapa crônica da parasitose devida a esse protozoário, considerando que ela é mais proeminente sob o ponto de vista médico-assistencial. A propósito, efetuamos apreciações baseadas em modelo composto por camundongos infectados e, também, relacionadas com indivíduos acometidos de doença de Chagas. Usamos 100 mg/kg/dia, por meio de sonda gástrica, durante três meses, e 100 ou 200 mg/dia, pela via oral, respectivamente, no decurso de igual período, sem evidenciarmos efeitos benéficos, pelo menos conforme a metodologia adotada

Immune response in mice immunized with acidic antigenic fractions from Trypanosoma cruzi cytosol

The humoral and cellular immune responses as well as the resistance to infection with bloodstream forms of T. cruzi were studied in mice immunized with acidic antigenic fractions from parasite cytosol, F III and F IV, plus Bordetella pertussis as adjuvant. The immunization with F III induced positive ITH and DTH responses to homologous antigens. In mice immunized with F IV, the ITH was negative and four out of six animals presented positive DTH reactions. In both groups of mice the analysis of IgG aginst T. cruzi showed that the major isotype elicited was IgG1. Specific IgE was also detected in sera from F III immunized mice, thus confirming the presence of homocytothropic antibodies. The parasitemias reached by F III and F IV immunized mice after challenge were lower than those of the controls showing in this way a partial protection against the acute infection. The histological studies of heart and skeletal muscle performed two months after the infection revealed variable mononuclear infiltration in all infected mice despite immunization.

Hemolymphatic components in vectors of Trypanosoma cruzi: study in several species of the subfamily Triatominae (Hemiptera: Reduviidae)

The members of the subfamily Triatominae (Hemiptera : Reduviidae) comprise a great number of species of medical importance in the transmission of the T. cruzi (American trypanosomiasis). The aim of this study was to contribute to the knowledge about the chemical composition in proteins, lipids, lipoproteins, and carbohydrates of vectors of Chagas' disease corresponding to twelve members of the subfamily Triatominae. This study was carried out in ninphs of the fifth instar and adult males of the species: T. delpontei, T. dimidiata, T. guasayana, T. infestans, T. mazzotti, T. pallidipennis, T. patagonica, T. platensis, T. rubrovaria, T. sordida of the Triatoma genus, and D. maximus and P. megistus of the Dipatalogaster and Panstrongylus genera respectively. The results show on one hand, qualitative differences in the protein composition, and on the other hand, similarity in the lipoprotein profiles. Lipids, proteins, and carbohydrates did not show significant differences between species or/and stages.

Trypanosoma cruzi meningoencephalitis and myocarditis in a patient with acquired immunodeficiency syndrome

We report the case of a 52-year-old male heterosexual patient with acquired immunodeficiency syndrome (AIDS) and reactivation of Chagas' disease manifested by meningoencephalitis and myocarditis, diagnosed post-mortem. Unexplained reactivation of Chagas' disease should be included among the diagnostic criteria of AIDS in human immunodeficiency virus positive patients. On the other hand, AIDS should be considered in the differential diagnosis of patients with unexplained reactivation of Chagas' disease.

Prevalencia de infeccion humana por Trypanosoma cruzi en bancos de sangre en Venezuela

Las primeras investigaciones realizadas a nivel de bancos de sangre, durante la década 50, indican que la seroprevalencia por infecciones a T. cruzi entre hemodadores fue de 12%. Un estudio posterior, entre 1963-64, efectuado en varios bancos de sangre, así como otros centros, registró una seroprevalencia global de 6.0% (1.1-10.1%). La donación de sangre en Venezuela es gratuita. El control de los bancos de sangre recae en el Departamento de Transfusiones y Bancos de Sangre del Ministerio de Sanidad y Asistencia Social. A partir de 1988, se emplea uniformemente la técnica de ELISA para el diagnóstico de infecciones a T. cruzi en los Bancos de Sangre. La seropositividad promedio interanual, entre 1984-1992, fue de 1.20% (1.09-1.94%). Existen variaciones geográficas entre las localidades de varias entidades federales. Los estados con mayor prevalencia se ubican en las regiones del occidente y centro del país, a saber: Portuguesa, Barinas, Lara, Trujillo, Cojedes y Carabobo. Por las dificultades en obtener tasas de incidencia para el Mal de Chagas, resulta adecuado emplear tasas de prevalencia para uso en salud pública, en función de su mayor estabilidad; y en el caso de Venezuela, dada la severidad menor y una sobrevivencia mayor por esta patologia hoy día. La especificidad, como parámetro de las pruebas serológicas, debería considerarse en función de la baja seroprevalencia detectada a nivel nacional. Convendría emplear varias pruebas diagnósticas en paralelo para buscar un equilibrio entre sensibilidad y especificidad.

Avaliação parasitológica e sorológica de caprinos inoculados experimentalmente pelo Trypanosoma cruzi

A infecção experimental pelo Trypanosoma cruzi foi estudada em nove caprinos jovens. Os animais foram inoculados via intraperitoneal com 10³ tripomastigotas sangüíneos/Kg de peso com as cepas 147 (Grupo 1) e 229 (Grupo 2), isoladas de pacientes chagásicos crônicos de Bambuí, MG. Realizaram-se exames de sangue a fresco, xenodiagnóstico, hemocultura e sorologia (RIFI e ELISA). O período de acompanhamento da infecção variou de 7 a 30 meses. Pôde-se observar uma acentuada diferença no curso da infecção entre os dois grupos. Demonstrou-se pela primeira vez em caprinos experimentalmente inoculados com T. cruzi, o parasita através de exames de sangue a fresco e xenodiagnóstico no período agudo e pela hemocultura e xenodiagnóstico no período crônico da infecção chagásica. Os resultados sugerem que caprinos, em determinadas condições epidemiológicas, podem ser importantes no ciclo de transmissão silvestre e peridoméstico do T. cruzi, especialmente aqueles animais mais jovens.