Resumo em Português:

Os autores passam em revista os diferentes antígenos até agora empregados na reação de fixação do complemento na moléstia de Chagas. Propõem o uso de um antígeno alcoólico feito com culturas de S. cruzi, assim preparado: lavam três vezes os flagelados das culturas com soro fisiológico; juntam acetona pura (10 vezes o volume do depósito) e deixam 24 horas agitando freqüentemente; centrifugam, desprezam o líquido e secam o depósito a 37°; pesam, trituram e juntam álcool absoluto na quantidade 1 cm³ por cada centigramo; conservam a 37° durante 20 dias, agitando diariamente; para uso, diluem o álcool límpido na água fisiológica aos poucos e sob constante agitação. A reação foi praticada com 83 soros humanos, sendo positiva em 96,3% dos casos de doença de Chagas e em 81,8% dos casos de leishmaniose tegumentar, doença cujo agente etiológico tem estreitas afinidades com S. cruzi. Em 81 soros a reação foi feita paralelamente com a R. W., obtendo-se apenas 4 vezes a positividade de ambos os testes. Os primeiros resultados das reações feitas com este antígeno podem ser considerados bastante favoráveis, sendo entretanto necessária uma maior experiência para se ajuizar do valor real da reação no diagnóstico da moléstia de Chagas.

Resumo em Português:

Culturas de tecido de embrião de galinha foram contaminadas com Schizotrypanum de diversas procedências. Serviram como material de contaminação, culturas em tubos de agar-sangue e em um caso sangue parasitado de cobaia. Os transplantes de tecidos foram mantidos em cultura de acordo com a técnica de Carrel, ligeiramente modificada. Os tecidos usados foram baço, miocárdio, fígado, epitélio da iris, gânglio espinhal e monócitos do sangue de galinha adulta. Em todos estes tecidos o Schizotrypanum foi facilmente cultivado, parasitando os diferentes tipos de células existentes: fibroblastos, histiocitos, macrófagos, células epiteliais, células nervosas, etc. Os autores puderam seguir o ciclo completo do parasito, confirmando os conhecimentos clássicos da sua evolução no tecido: transformação em leishmânia, multiplicação por divisão binária, transformação em critídia e finalmente em tripanosoma adulto. Também observaram formas parasitárias que aparentemente evoluiram diretamente da forma de leishmânia à de tripanosoma, sem passar por critídia. Conseguiu-se a infecção de um camondongo inoculado com S. cruzi de origem humana passado por cultura de tecidos. Os autores também estudam diferentes fenômenos observados nas relações entre as células e os parasitos.

Resumo em Inglês:

Chick embryo tissue cultures were smeared with Schizotrypanum from different sources. The cultures were inoculated with flagellates from blood-agar cultures and in one instance from blood of an infected guinea-pig. Carrel’s technique of tissue culture, with slight modifications, was employed. The tissue used were spleen, myocardium, liver, epithelium of the iris, spinal ganglion and monocytes from chicken blood. In all these tissues the flagellate developed easily, parasitizing different types of cells: fibroblasts, histiocytes, macrophages, epithelial cells, cells of the nervous system, etc. The authors were able to follow the complete cycle of the parasite, observing its classic evolution: transformation into leishmania, multiplication by binary division, transformation into crithidia and finally into trypanosome. They also observed forms of the parasite which possibly developed directly from leishmania to trypanosome without apparently going through the crithidia stage. They succeeded in infecting a white mouse with a human strain of S. cruzi after passage through tissue culture. The authors also pointed out different phenomena observed in the relations between the cells and the parasites.

Resumo em Inglês:

Last October 2nd the Author smeared nine tubes of Loewenstein medium with material obtained from closed pustulae of a seven years old boy, L2 case of leprosy. This material was very rich in Hansen bacilli in its different forms, inclusive globus, as is seen in the figures 2 and 3 of Plate 1. Part of this material obtained from pustulae opened by galvanocautery, was inoculated, at the same day, into white rats and guinea-pigs. November 26th a new biopsy gave more rich material, which was smeared again into Loewenstein fresh medium. December 15th three of the first and two of the second series of tubes of cultures showed germination of a yellow, dry and rough culture, covering almost the total surface of the medium. Microscopic examination of the culture showed that it was a pure culture of an acid-fast organism. Passages into glycerinated potatoes germinated well covering the surface of the same with a clear yellow granulated culture remaining the fluid (glycerinated water) quite limpid. The germination in glycerinated broth produced a yellow velum in the surface of the medium, as is seen in fig. 3 of Plate 2, without becoming turbid. The microorganism isolated twice from the same source of material was cocciforme (as Mycobacterium pulviforme of Marchoux), in the original culture, becoming more bacilliforme, always acid-fast, after passage into glycerinated media. The A. sent his culture to foreigner colleagues to study it and will inoculate it soon into laboratory animals.

Resumo em Inglês:

The first agglutination experiments (Tables 1 and 2) showed that the serum obtained with any one strain of Leishmania, agglutinates all the others even of another species. This finding reveals the existence of a common antigen. However as the titre of agglutination did not permit a sharp differentiation of species we tried the adsorption method. The first adsorption tests made demonstrated differences in antigenic constitution between a strain of. L. donovani on one hand and strains of L. tropica or L. brasiliensis on the other. Further experiments in which L. chagasi was tested against the other species revealed that the former was antigenically different from the others. These tests were performed by adsorbing an anti-chagasi serum with organisms belonging to the other species or, conversely, adsorbing with L. chagasi sera prepared against the other species (See Tables 9 to 24). On the other hand, the adsorption of a serum prepared against one strain of l. chagasi by another of the same species showed that they had identifical antigenie constitution. These findings suggested the possibility of separating different species of Leishmania by this method. However, tests to separate the other species from one to another gave inconclusive results. (See Tables 27 to 35). It was soon observed that all the strains of L. chagasi were of recent isolation while all the others had been maintained in artificial culture media for a long time. We were led to believe that this condition was responsible for the differences in behaviour encountered. Accordingly, recently isolated strains of L. brasiliensis and L. donovani were tested and shown to be antigenically similar to strains of L. chagasi also recently isolated. The conclusion may be drawn that all strains have the same antigenic constitution when freshly isolated. It has been noted that when a serum which has been prepared against a freshly isolated is adsorbed with an old strain, the amount of agglutinins left free, is much smaller than when a serum prepared against an old strain is adsorbed with a newly isolated strain. At first, we thought to explain this by the low titre of the serum. However, the amount of agglutinins left free was not larger when higher titre serum was tested. The results do not corroborate the view of a special antigen being present in recently isolated strains (vi antige) but rather that the phenomenon is dependent on differences of the amount of the common antigen, more abundant in recent strains. In order to make this clear, experiments were made in which equal amounts of a serum prepared against a newly isolated strain were adsorbed by equal amounts, by weight, of, on one hand, a new strain, and the other an old strain. The resulting adsorbed sera were then titrated. (Table 44). Results showed that newly isolated strains adsorb a larger amount of agglutinins (Tables 44, 45). Two hypothesis have bem advanced to explain the stronger adsorbing qualities of the newly isolated strains. 1° - these strains possess larger amounts of the common antigen and 2° - they contain a vi antigen which adsorbed by the new strain together with the common antigen is the cause of their larger adsorbing capacity. To find out which of the two hypothesis corresponds to the reality a new experiment was made, similar to the one summarized in table 44. The adsorbed sera were made to act on a recently isolated strain as well as on an old one. The latter, not containing the vi antigen, the difference seen when sera act on new strains should not be observed here in the case of this antigen being responsible for the differences in adsorbing properties. The difference persisting, the indication would be that the greater adsorbing capacity of recently isolated strains was really related to larger amounts of the common antigen present (Tables 46 and 47). The results of the experiment excluded the possibility of the vi antigen being responsible. Other experiments, (Tables 48 to 53) using a 3 year old strain, demonstrated the modification in its antigenic constitution during the period it was maintained in cultures.

Resumo em Português:

1) Em trabalho sistematizado, foi estudada pela primeira vez a epidemiologia da doença de Chagas, em um trecho da região do estuário do Amazonas, cujas condições ecológicas diferem das encontradas em outras zonas onde teem sido feitas pesquisas semelhantes. 2) No local escolhido para estudo – Aura – uma localidade distante cerca de 10 km. de Belém, (Pará), não foi encontrada nenhuma infecção humana pelo S. cruzi, quer pelo exame de sangue, quer pelo xenodiagnóstico, ambos feitos em todos os habitantes (117 indivíduos). Não foi encontrada também sintomatologia atribuível à doença. 3) Em 6 xenodiagnósticos feitos em animais domésticos (5 cães e 1 gato), foi encontrado um cão infectado. Este animal pertencia á casa onde foi verificada maior infestação por Triatomídeos, com índice de infecção elevado. Pelas razões expostas no texto, porem, foi sugerida a hipótese do mesmo se ter infectado pela ingestão de vísceras de animais silvestres contaminados. 4) De 115 animais silvestres, cujo sangue foi examinado em gota espessa, 9 mostraram-se parasitados por Schizotrypanum (7,8%). Em 47 verificações pelo xenodiagnóstico, 15 animais silvestres (11 mucuras, 3 tamanduás e 1 tatú) se apresentaram positivos (32,6%) . O tamanduá (T. tetradactylus) foi pela primeira vez assinalado como depositário natural do Schizotrypanum. 5) Como resultado de buscas feitas durante 11 meses, sendo cada domicílio inspecionado pelo menos de 15 em 15 dias, foram encontrados Triatomídeos em 7 das 36 casas existentes no povoado. Todos os 39 exemplares capturados eram adultos; a procura exaustiva não revelou a existência de formas jovens no interior das habitações. Na ordem de freqüência, as espécies encontradas foram as seguintes: P. geniculatus (29 exemplares, dos quais 7 infectados); R. pictipes ( 9 exemplares, dos quais 2 infectados); E. mucronatus ( 1 exemplar, não infectado) . A grande maioria dos insetos foi capturada na segunda metade do ano (época do estío). 6) Em plena mata, n’uma toca de tamanduá (T. tetradactylus) foram encontradas larvas, ninfas e adultos de P. geniculatus. A casa situada mais próximo desse foco, foi a que apresentou maior infestação (22 exemplares) e exclusiva para essa espécie. Em uma toca de macaco da noite (P. flavus), foi encontrada uma larva de Triatomídeo. Este foco também ficava próximo à casa acima referida. Em toca de P. flavus foi também achado um exemplar adulto de Panstrongylus refotuberculatus. 7) Amostras de S. cruzi isoladas de animais silvestres, mostraram fraco poder infectante. A amostra isolada do cão, embora infectando facilmente os animais de laboratório, pelos estudos biométricos feito por DIAS e FREITAS, afasta-se das amostras humanas típicas. 8) São discutidos os resultados acima referidos e, pelos hábitos dos transmissores, pela predominância de depositários silvestres do parasito, conclue-se pela natureza silvestre da Tripanosomiase Americana no local estudado. Se bem que não tenham sido verificadas infecções humanas, dado o encontro de um cão parasitado – infecção esta que se pode ter verificado pelo meio normal da transmissão da moléstia – admite-se a possibilidade do aparecimento de casos humanos nessa região. Ressalta-se a confirmação que tais resultados parecem trazer á hipótese de CARLOS CHAGAS, que pensava ser esta doença primitivamente silvestre, com posterior adaptação aos animais domésticos e ao homem.

Resumo em Inglês:

1) The epidemiology of Chagas’ disease was studied for the first time in a systematized work, in a district of the region of the Amazon estuary, whose ecological conditions differ from those encountered in other zones where similar researches have been made. 2) At the place chosen for the studies – Aurá – a locality about 10 kms from Belem (State of Pará), no human infection by S. cruzi has been found, either by blood examination or xenodiagnosis, both having been tried on all the inhabitants (117 people); nor was there any symptomatology attributable to the disease. 3) Out of the six xenodiagnoses made on domestic animals (five dogs and one cat), only one dog was found infected. This animal belonged to the house where the greatest infestation by triatomid bugs with a high index of infection was verified. However, for the reasons given in the text, the hypothesis that the dog might have become infected by eating organs of infected wild animals has been suggested. 4) Of the 115 animals whose blood was examined in thick blood film, 9 (7,8%) proved to be infected by Schizotrypanum. Out of 47 xenodiagnoses carried out on wild animals, 15 (32.6%) showed positive results (11 opossums, 3 ant-eaters, 1 armadillo). The ant-eater T. tetradactylus is a new vertebrate-host of Schizotrupanum. 5) During 11 months of research, each house being inspected at least once every fortnight, triatomid bugs were found in 7 out of the 36 houses of the village. The 39 specimens collected were all adults; an exhaustive search did not reveal the existence of any young insects in the houses. According to their order of frequency, the species found were the following: P. geniculatus (29 specimens, 7 infected), R. pictipes (9 specimens, 2 infected) and E. mucronatus (1 uninfected specimen). The great majority of the insects was captured during the second half of the year (summer period). 6) In the very midst of the forest, in the hole of an ant-eater (T. tetradactylus), larvae, nymphs and adults of P. geniculatus were found. The house nearest to the hole presented the highest index of infestation by this bug (22 specimens, all in adult stage). In the hole of Potus flavus (“macaco da noite”) a triatomid larva and an adult Panstrongylus rufotuberculatus have been found. 7) Strains of Schizotrypanum isolated from wild animals showed a weak virulence. The dog strain, although easily infecting laboratory animals, differs from the typical human strains, as DIAS and FREITAS proved in their biometrical studies. 8) The results above mentioned are discussed in the paper and, in view of the habits of the insect vectors and the prevalence of infected wild animals, the sylvan nature of Chagas’s disease in Aurá is concluded. Although human cases have not been detected, the possibility of human infection in this region is admitted, since a domestic animal (a dog) was found to be infected (possibly through the way of the insect vector). Attention is drawn to the confirmation such results seem bring CARLOS CHAGAS” hypothesis, that this disease was at first found in wild animals, and could later on spread to domestic animals and man.