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.

Alkaline soluble Trypanosoma cruzi epimastigote antigen (ASEA) applied to dot-ELISA

The alkaline soluble Trypanosoma cruzi epimastigote antigen (ASEA) was assessed in dot-ELISA for the diagnosis of Chagas' disease. Serum samples (355) from chagasic and non-chagasic patients were studied, and IgG antibodies to ASEA were found in all patients with chronic Chagas' disease. In non-chagasic patients 95.6% were negative, except for those with leishmaniasis (visceral and mucocutaneous), and some patients from control group reacted in low titers. The data indicate that dot-ELISA using ASEA is suitable for seroepidemiologic surveys to be employed in endemic areas for Chagas' disease.

Francine Benoit e a cultura musical em Portugal : estudo das críticas e crónicas publicadas entre 1920- e 1950

Dissertação de Mestrado em Ciências Musicais

Murine virus contaminant of Trypanosoma cruzi experimental infection

The possibility that some virus contaminants could be altering host response to Trypanosoma cruzi experimental infection was investigated. Data obtained showed that CBA/J mice infected with stocks of parasite maintained in mice (Y1UEC) presented higher level of parasitemia and shorter survival times than those infected with a stock (Y1TC) which was also maintained in mice but had been previously passaged in cell culture. Mouse antibody production tests, performed with the filtered plasma of mice infected with Y1UEC, indicated the presence of mouse hepatitis virus (MHV) while no virus was detected when testing the plasma of Y1TC infected mice. Filtered plasma of Y1EUC infected mice was shown to contain a factor able to enhance the level of parasitemia and to reduce the mean survival time of mice challenged with 10(5) Y1TC. This factor, that could be serially passaged to naïve mice was shown to be a coronavirus by neutralization tests.

Investigation on the possibility of spontaneous cure of mice infected with different strains of Trypanosoma cruzi

Seventy Swiss mice chronically infected with different strains of Trypanosoma cruzi, with persistently negative parasitemia on routine blood examination were parasitologically investigated to find out whether spontaneous cure occurred. Duration of infection varied from 90 to 250 days in the initial phase of this investigation. Parasitological tests consisted of daily direct blood examination performed during at least 25 days, followed by xenodiagnosis and subinoculation of blood into newborn mice. Mice that persisted negative were treated with Cyclophosphamide with one dose of 250 mg/kg of body weight and then investigated by direct blood examination, xenodiagnosis and subinoculation. A second dose of 250 mg/kg b. w. was given to the persistently negative mice. With one single exception, all mice showed positive parasitological tests in the different stages of the present investigation and we conclude that spontaneous cure did not occur in this group, which is representative of the chronic infection with different strains of T cruzi.

Characterization of a Trypanosoma cruzi strain isolated from a non-endemic area in Northeast Brazil

A cepa WSL (Wild São Lorenço) de T. cruzi, isolada de um cobaio proveniente de São Lorenço da Mata (Nordeste do Brasil) foi caracterizada através da análise do seu comportamento morfobiológico e perfil isoenzimático. Para o estudo do comportamento morfobiológico, tripomastigotas sanguíneos (1 x 10 5) da cepa WSL foram inoculados por via intraperitonal em camundongos albinos Swiss. Como controle a cepa Y (Tipo I) foi usada. Durante o curso da infecção os seguintes parâmetros foram analisados: parasitemia, mortalidade, morfologia dos parasitas no sangue periférico e tropismo tissular. O perfil isoenzimático foi analisado em relação às enzimas ALAT, GPI e PGM usando como controle de referência as cepas Peruana (Tipo I), 21SF (Tipo II) e Colombiana (Tipo III). A cepa WSL apresentou as seguintes características biológicas: 1) multiplicação lenta e pico parasitêmico entre 21 - 25 dias pós-infecção; 2) mortalidade de 3,3% 40 dias pós-infecção; 3) predominância de formas largas no sangue periférico e 4) miotropismo com predominante envolvimento cardíaco. A análise isoenzimática mostrou um padrão de zimodema 2 (Z2) que corresponde às cepas biológicas Tipo II. Os resultados mostram que a cepa WSL apresenta baixa virulência e patogenicidade.

Trypanosoma cruzi strains: behavior after passage into authoctonous or foreign species of triatomine (biological and biochemical patterns)

The behavior of T. cruzi strains from S. Felipe - BA (19 SF, 21 SF and 22 SF) classified as Type II Zymodeme 2, was investigated after passage through the authoctonous (P. megistus) and foreign vectors (T. infestans and R. prolixus). For each strain Swiss mice were infected: I - with blood forms (control); II - with metacyclic forms (MF) from P. megistus; III - with MF from T. infestans; IV - with MF from R. prolixus. Inocula: MF from the three species of triatomine, 60 to 120 days after feeding in infected mice, adjusted to 10 4. Biological behavior in mice (parasitemia, morphology, mortality, virulence and pathogenicity) after passage through triatomine was compared with data from the same strain in control mice. Isoenzymic electrophoresis (ASAT, ALAT, PGM, GPI) were also performed after culture into Warren medium. The three strains maintained the isoenzyme profiles (zymodeme 2), in the control groups and after passages through different species of triatomine. Biological characterization disclosed Type II strains patterns for all groups. An increased virulence was observed with the 22 SF strain isolated from P. megistus and T. infestans and higher levels of parasitemia and predominance of slender forms in mice inoculated with the 19 SF and 21 SF from these same species. Results indicate that the passage through the two species T. infestans and P. megistus had a positive influence on the virulence of the regional strains of S. Felipe, regardless of being autocthonous (P. megistus) or foreign to the area (T. infestans).

Etiological drug treatment of human infection by Trypanosoma cruzi

Forty-nine American Trypanosomiasis (Chagas' disease) patients, with xenodiagnosis proven parasitemia were treated by the authors. Forty-one of these patients were given benznidazole, at dosages ranging from 5mg/kg/day to 8mg/kg/day, during a pre-established period of 60 days. In this group, 17 patients had an undetermined form of the disease, whereas 22 had cardiologic disease and 4 had digestive disease (two patients had a mixed form of the disease). Side effects were frequent, and led to the discontinuation of treatment in 17 patients. The follow-up period ranged from 1 to 20 years (mean follow-up period of 6 yrs. 7 mo). 26 (63.4%) of the patients became parasitemia-negative. The other eight patients were treated with nifurtimox, during 120 days, following a variable dose regime of 5mg/kg/day (initial dose) to 17 mg/kg/day (final dose). Six of them had severe side effects, and only one patient remained parasitemia-negative throughout the observation period (ranging from 1 to 18 years). Benznidazole proved to be better tolerated and more effective in the management of parasitemia when compared to nifurtimox, although more effective and less toxic drugs are still desirable.

Cross-reactivity of antibodies in human infections by the kinetoplastid protozoa Trypanosoma cruzi, Leishmania chagasi and Leishmania (Viannia) braziliensis

We have detected antibodies, in the sera of Chagas disease, Kala-azar and Mucocutaneous leishmaniasis patients, that bind multiple antigens shared between the three causative agents. The Chagas disease sera showed 98 to 100% positive results by ELISA when the Leishmania braziliensis and Leishmania chagasi antigens were used, respectively. The Kala-azar sera showed 100% positive results with Trypanosoma cruzi or L. braziliensis antigens by immunofluorescence assays. The antibodies in the sera of Mucocutaneous leishmaniasis patients showed 100% positive results by ELISA assays with T. cruzi or L. chagasi antigens. Furthermore, the direct agglutination of L. chagasi promastigotes showed that 95% of Kala-azar and 35% of Mucocutaneous leishmaniasis sera agglutinated the parasite in dilutions above 1:512. In contrast, 15% of Chagas sera agglutinated the parasite in dilutions 1:16 and below. Western blot analysis showed that the Chagas sera that formed at least 24 bands with the T. cruzi also formed 13 bands with the L. chagasi and 17 bands with the L. braziliensis. The Kala-azar sera that recognized at least 29 bands with the homologous antigen also formed 14 bands with the T. cruzi and 10 bands with the L. braziliensis antigens. Finally, the Mucocutaneous leishmaniasis sera that formed at least 17 bands with the homologous antigen also formed 10 bands with the T. cruzi and four bands with the L. chagasi antigens. These results indicate the presence of common antigenic determinants in several protozoal proteins and, therefore, explain the serologic cross-reactions reported here.

Seropositivity for anti-Trypanosoma cruzi antibodies among blood donors of the "Hospital Universitário Regional do Norte do Paraná", Londrina, Brazil

The most frequent form of acquisition of Chagas' disease in endemic areas was the transmission through the feces of contaminated triatominae. However, special attention should be paid in urban areas to transmission by blood transfusion, justifying the compulsory screening of blood donors. Early investigations at blood banks in the town of Londrina, Brazil, demonstrated that the seroprevalence of anti-Trypanosoma cruzi antibodies among blood donors was approximately 7.0% in the fifties9,34. Further studies demonstrated pratically the same seroprevalence until the eighties4,32,41. In an attempt to obtain data about the real dimension of the seropositivity for anti-Trypasonoma cruzi antibodies in the region, the authors carried out a large-scale study on 45,774 serum samples from blood donors of the Hemocentro of Hospital Universitário Regional do Norte do Paraná (HURNP), Universidade Estadual de Londrina. The immunological tests were done at the Division of Clinical Immunology of HURNP from May 1990 to December 1994. The serum samples were studied by the indirect hemagglutination assay (IHA, using kits commercially obtained from EBRAM) and by indirect immunofluorescence (IFI, using kits from LIO SERUM) with anti-human IgG conjugate (LABORCLIN). The results demonstrated that 643 serum samples were positive in both assay corresponding to a seroprevalence of 1.4%, i.e., a significant decrease in anti-Trypanosoma cruzi antibodies in the region in comparison with the previously mentioned rates. Data correlating sex and age of seropositive blood donors are presented, as well as the possible factors that may have contributed to the results observed.

Infection by Trypanosoma cruzi in mammals in Yucatan, Mexico: a serological and parasitological study

In order to determine Trypanosoma cruzi infection among mammals in Yucatan, Mexico, 372 animals, both wild and synanthropic including carnivores, marsupials and rodents were studied. Serological studies by indirect haemagglutination (IHA) were carried out to detect antibodies to T. cruzi and a parasitological study was also performed (blood smear and histopathology). Of all the animals tested 18.54% were serologically positive, with a significantly higher frequency among the wild ones (33.33%) compared to the synanthropic ones (17.79%). To determine T. cruzi in positive animals, blood was inoculated into a white mouse (webster type) to prove myocardium colonization. The serological and parasitological positivity of these animals, as well as their behavior in the environment, taken together with the socioeconomic and cultural characteristics of the population, suggest that in Yucatan, Mexico, Canis familiaris, Didelphis marsupialis and Rattus rattus act as a link with the wild cycle.

Human infection with Trypanosoma cruzi in Nasca, Peru: A seroepidemiological survey (1)

We estimated the proportion of seropositivity for infection with Trypanosoma cruzi (Chagas’ disease) in a sample of the rural population of the Province of Nasca, Department of Ica, southwestern Peru. Although Triatoma infestans, the only vector species identified in the Department of Ica, is often found in domestic environments, data of the extent of human infection with T. cruzi are scant. This study comprised 446 houses, known to be infested with triatomines, distributed in 19 rural localities. While visiting those houses we collected filter paper bloodspots from 864 occupants (of both sexes, aged one year or over). By means of the indirect fluorescent antibody test (IFAT), we detected anti-T. cruzi IgG antibodies in samples from 178 individuals (20.6%). Seropositivity was significantly more frequent in females (23.8%) than in males (17.5%). Among the 410 individuals in the 1- to 10-year-old age group (47.5% of the population sample), 85 (20.7%) were found seropositive, which is indicative of an early acquisition of the infection. Within this group no significant differences in seropositivity were associated with sex

Amastigotes forms of Trypanosoma cruzi detected in a renal allograft

Trypanosoma cruzi, the causative agent of Chagas’disease assumes two distinct forms in vertebrate hosts: circulating trypomastigote and tissular amastigote. This latter form infects predominantly the myocardium, smooth and skeletal muscle, and central nervous system. The present work describes for the first time the detection of amastigote forms of T. cruzi in the renal parenchyma of a kidney graft recipient one month after transplantation. The patient was serologically negative for Chagas’disease and received no blood transfusion prior to transplant. The cadaver donor was from an endemic area for Chagas’disease. The recipient developed the acute form of the disease with detection of amastigote forms of T. cruzi in the renal allograft biopsy and circulating trypomastigote forms. The present report demonstrates that T. cruzi can infect the renal parenchyma. This mode of transmission warrants in endemic areas of Chagas’disease