Use of saprophytic leptospira strains in the serodiagnosis of experimental leptospirosis in guinea-pigs (Cavia sp)

The efficiency of four Leptospira biflexa strains (Buenos Aires, Patoc 1, Rufino and São Paulo) as single antigen in the serodiagnosis in guinea-pigs experimentally infected with seven Leptospira interrogans serovars (canicola, grippotyphosa, hardjo, icterohaemorrhagiae, pomona, tarassovi and wolffi) was evaluated by the microscopic agglutination test. The four saprophytic strains were not able to reveal antibody titres in sera of guinea-pigs experimentally infected with Leptospira interrogans. Serological cross-reactions were observed between strains Patoc 1 and São Paulo and between serovars wolffi and hardjo.

Lytic antibodies elicited by Trypanosoma cruzi infection recognize epitopes present on both bloodstream trypomastigote and epimastigote forms of parasite

Sera of Chaga's disease patients containing anti-T. cruzi lytic antibodies were submitted to affinity chromatography using Sepharose 4B conjugated with antigen extracted from epimasiigote or trypomasiigote forms of the parasite. Epimastigotes were obtained from culture at the exponential growth phase and the trypomastigotes from blood of infected and immunosuppressed mice. Antigen of both parasite forms was obtained by sonication of the parasites followed by centrifugation. Both antigens were then conjugated to activated Sepharose 4B. Affinity chromatography was performed by passing sera from chagasic patients through an immunoadsorbent column containing either epimasiigote or trypomasiigote antigens. Antibodies bound to the column were eluted with cold 0,2 M glycine buffer pH 2,8. The eluted antibodies were analysed regarding their isotype and lytic activity. The results showed that anti-T. cruzi lytic antibodies present in sera from chagasic patients are mainly located in the IgG isotype and recognize epitopes present in both trypomasiigote and epimastigote forms. A brief report of this work has already been published12.

Effect of scorpion toxin on the enterochromaffin-like cells in normal and Trypanosoma cruzi-infected rats: a morphological study

Intravenous injection of scorpion toxin (Tityus serrulatus) in normal and Trypanosoma cruzi infected rats did not cause ultrastructural morphologic changes on enterochromaffin-like (ECL) cells of the stomach, although it induced a significant increase of the gastric secretion. Our data seem to indicate that gastric ECL cells structure is not affected by stimulation with scorpion toxin or by acute infection with T. cruzi in the rat.

Natural Trypanosoma cruzi infection in dogs of endemic areas of the Argentine Republic

The population dynamics and the prevalence of chagasic infection of 352 dogs living in 108 rural houses infested by triatomines were studied. The region was divided into three sections according to increasing distances to an urban area. Each animal was identified by means of its particular characteristics and built, and its owners gave information about its habits. By means of xenodiagnosis, serology and ECG studies, prevalences of infection, parasitological-serological correlation, percentage of altered electrocardiographic outlines and percentage of houses with parasitemic dogs, were determined. The rural area showed a characteristic T. cruzi infection pattern and differences in the canine population parameters with respect to the other areas were observed: a higher proportion of puppies than adult dogs, a more sedentary population, higher prevalences of infection, as measured by xenodiagnosis, in dogs, and the highest proportion of bedroom insects infected with T. cruzi. It is assumed that the sedentary characteristics of the human population in that rural area impinge in the blood offer to the triatomine population, and the high percentage of parasitemic dogs of the area, contribute to the rise of "kissing ougs" infected with T. cruzi found in bedrooms.

Estudo sorológico de infecções experimentais por Trypanosoma evansi, em cobaias

Comparamos os métodos de Imunofluorescência Indireta (IFI), Imunodifusão Radial Dupla e Aglutinação, para a pesquisa de anticorpos em soros, na tripanossomíase experimental por Trypanosoma evansi, em cobaias. Foram obtidas 20 amostras de soro correspondentes às 4 primeiras semanas de infecção. A IFI foi positiva em apenas 6 animais, com títulos variando de 1:4 a 1:16. Os títulos mais altos foram observados na 3ª semana pós-infecção. Anticorpos aglutinantes foram observados a partir da 1ª semana pós-infecção e, após a 2ª semana, todos os animais apresentaram reação de aglutinação positiva, com títulos variando de 1:8.000 a 1:250.000. O tratamento dos soros com 2-Mercapto-etanol inibiu a reação de aglutinação, sugerindo ser IgM a principal classe dos anticorpos presentes no soro dos animais infectados. Não se constatou a presença de anticorpos precipitantes durante todo o curso da infecção.

Differences in the antigenic profile of bloodstream and cell culture derived trypomastigotes of Trypanosoma cruzi

A comparative study of the antigenic profile of bloodstream and cell culture derived trypomastigotes showed many differences in their components. Using mouse anti-T. cruzi antibodies the differences were located mostly in the 120 kDa band, whereas using chagasic patient sera the differences were located in the 85 and 52 kDa bands. These findings might explain known physiological differences between trypomatigotes obtained from cell culture and from infected blood. A brief report of this work has already been published9.

Prevention of electrocardiographic and histopathologic alterations in the murine model of Chagas' disease by preinoculation of an attenuated Trypanosoma cruzi strain

The effects of infection with Trypanosoma cruzi on the electrocardiographic tracings of mice were studied in 4.groups of animals: (1) normal; (2) infected with a pathogenic T. cruzi strain (TS COB); (3) immunized with 3 intraperitoneal inocula of 10(6) attenuated T. cruzi epimastigotes (TCC) and (4) immunized-infected, which sequentially received the treatments of groups 3 and 2. Infection and protection were confirmed by xenodiagnosis and histopathology. Isolated alterations such as extrasystolia, 1st degree atrioventricular block, arrhythmia and ST elevation were observed in normal as well as infected mice. However, tracings taken repeatedly on each mouse over a 293 day period revealed a set of alterations which were more frequently seen in infected (14/22) than in normal (4/27) animals (p = 0.00048). These alterations consisted of supraventricular tachycardia, sinus bradycardia and persisting, first degree AV blocks, often associated to pacemaker changes. Inoculation of attenuated T. cruzi (group 3) did not increase these alterations (2/27 mice) but significantly prevented their development after challenge with the pathogenic strain (1/19 versus 14/22 mice, p = 0.000095). Thus, preimmunization reduced not only parasitemia but also a pathogenic consequence of T. cruzi infection. This evidence is relevant for immunoprevention studies against Chagas' disease.

Incidência de escherichia coli enterotoxigênica (ETEC), rotavirus e Clostridium perfringens de casos de diarréia em crianças, na região de Campinas, SP, Brasil

Foi realizada uma pesquisa na região de Campinas, SP, Brasil, sobre a presença de Escherichia coli enterotoxigênica (ETEC), rotavírus e Clostridium perfringens enterotoxigênico em fezes diarréicas de crianças com até 2 anos de idade. Dos 132 espécimens fecais examinados quanto à presença de ETEC 27 (20,45%) foram positivos. Destes foram isoladas 41 amostras de ETEC, das quais 40 produziram apenas a enterotoxina termolábil (LT) detectada pelo teste de imuno hemólise radial modifi cado. Entre as 183 amostras de fezes examinadas para rotavírus, 29 (15,84%) foram positivas pelas técnicas de eletroforese em gel de poliacrilamida (PAGE) e ensaio imunoenzimático (EIE), sendo que destas, 15 (51,7% ) foram provenientes de materiais coletados nos meses de inverno. Todas as amostras pertenciam ao grupo A e, através da técnica de PAGE, pode-se observar que o tipo eletroforético mais freqüente (9 amostras) foi designado Ib, IIc, Illb, IVa, de acordo com a classificação por nós adotada. Apenas 113 amostras de fezes foram examinadas para a presença de C. perfringens enterotoxigênico. Para a detecção da enterotoxina nos sobrenadantes das culturas foram utilizadas as técnicas de hemaglutinação passiva reversa e inoculação intravenosa em camundongos, sendo encontradas 12 (10,61%) amostras entero-toxigênicas. Diante destes resultados é chamada a atenção sobre o valor apenas relativo de uma coprocultura convencional para fins de diagnóstico, ressaltando-se a importância da criação de métodos simplificados que favoreçam a detecção e identificação dos grupos de agentes enteropatogênicos estudados na presente pesquisa.

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.

Human parasitism by Phagicola sp (Trematoda, Heterophyidae) in Cananéia, São Paulo State, Brazil

We report one case of parasitism by Phagicola sp. (Trematoda, Heterophyidae) in a 31 years-old woman who, in 1987, travelled and stayed several months in the municipality of Cananéia (SP), where she ingested, in various occasions, raw mullet (Mugil sp.). The patient refered mild intestinal pain and laboratory examinations showed eggs of Phagicola sp. in the stools and a slight increase in eosinophil blood levels (8%). After treatment with praziquantel (75 mg/kg per day for three days) all the symptoms and signs disappeared. This is, certainly, the first record of human infection by Phagicola sp. in Brazil and, perhaps, in countries other than the U.S.A. where unclear references to a few human cases were reported in the South-eastern region.

Isolation of Leishmania sp. from aqueous humor of a patient with cutaneous disseminated leishmaniasis and bilateral iridocyclitis (preliminary report)

The Authors report an uncommon case of leishmaniasis with disseminated cutaneous lesions, systemic manifestations and ocular involvement, the latter being characterized by bilateral nongranulomatous iridocyclitis. The severity of the oph-thalmologic lesions and its unresponsiveness to therapy (in spite of satisfactory regression of both systemic and cutaneous manifestations) lead to a needle aspiration of the anterior eye chamber, content. From this material Leishmania sp was isolated. To our knowledge this is the first time that Leishmania has been shown into the ocular globe.

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.

Determination du niveau d'anticorps antitetaniques en donneurs de sang au moyen du test ELISA-TÉTANOS (São Paulo, SP, Bresil)

Le test ELISA-TÉTANOS (Biosys, France) a été utilisé pour le titrage des anti-corps tétaniques (sensibilité = 0,0025 UI/ml) en sérums humains de donneurs de sang, 566 hommes et 108 femmes, âges de 18 à 58 ans, moyenne de 29 ans, provenant de São Paulo, SP, Brésil. L'OMS, acceptant seulement la séroneutralisation sur souris (NT), la méthode de référence, pour les études sur la protection contre le tétanos, préconise le titre de 0,01 UI/ml comme minimum protecteur. BOURLEAUD & HUET ont proposé la limite de 0,06 UI/ml quand s'emploie le test ELISA, en attendant à une certaine discordance inévitable entre les méthodes. Parmi les 674 sérums étudiés, 178 (26,41%) n'ont pas présenté d'anticorps (< 0,0025 UI/ml); 413 (61,28%) ont présenté des résultats égaux ou supérieurs à 0,01 UI/ml et en 232 (34,42%) les titres ont été égaux ou supérieurs à 0,06 UI/ml. Le pourcentage d'individus protégés a été inversement proportionnel à 1'âge: environ 50% dans le groupe le plus jeune (hommes de 18 à 35 ans et femmes de 18 à 23 ans) contre environ 10% dans le groupe de plus de 42 ans ont présenté des titres sûrement protecteurs (> 0,06 UI/ml).