Characteristics of Triatomine infestation and natural Trypanosoma cruzi infection in the State of Rio Grande do Norte, Brazil

Abstract: INTRODUCTION Natural and artificial ecotope infestation by the kissing bug triatomines and their colonization and infection by Trypanosoma cruzi , the Chagas disease agent, were evaluated in nine municipalities of the State of Rio Grande do Norte, Brazil. METHODS Following identification, triatomine intestinal contents were analyzed by direct microscopic examination, xenoculture, and polymerase chain reaction (PCR) for parasite detection. Trypanosoma cruzi isolates were genotyped using three different markers. RESULTS Of 842 triatomines captured, 65% were Triatoma brasiliensis , 17.8% Triatoma pseudomaculata , 12.5% Panstrongylus lutzi , and 4.7% Rhodnius nasutus . Triatoma brasiliensis and P. lutzi adults were found in the intradomicile. T. brasiliensis, T. pseudomaculata , and R. nasutus nymphs and adults were found in the peridomicile and wild environment. Intradomiciliary and peridomiciliary infestation indexes were 5.6% and 33.7%, respectively. In the peridomicile, chicken coops were the most infested ecotope. The T. cruzi triatomine infection rate was 30.2%, of which PCR detected 29%. P . lutzi (78.1%), T . brasiliensis (24.5%), and T . pseudomaculata (22.7%) were the most infected species. TcII and III genotypes were detected in T. brasiliensis and TcIII in P. lutzi . CONCLUSIONS T. brasiliensis was found in all environments and most ecotopes with high T. cruzi infection rates. High infection rates were also detected in T . pseudomaculata and P. lutzi , suggesting their role in the interchange between the wild and peridomestic transmission cycles. The combination of PCR, microscopic examination, and xenoculture contributed to improving T. cruzi infection evaluation in triatomine bugs. The TcII and TcIII genotypes were predominant in the study area.

Trypanosoma cruzi isolated from a triatomine found in one of the biggest metropolitan areas of Latin America

Abstract: INTRODUCTION: To characterize Trypanosoma cruzi (TcI) isolated from a Panstrongylus megistus specimen found in one of the biggest metropolitan areas of Latin America, the relationship between the TcI group of T. cruzi and the transmission cycle in the urban environment was studied. METHODS: The T. cruzi strain, Pm, was isolated in a culture medium from the evolutionary forms present in the hindgut of a live male specimen of P. megistus found in the Jabaquara subway in São Paulo City. The sample from the triatomine showed trypomastigote forms of Trypanosomatidae, which were inoculated in the peritoneum of Balb/c mice. The sample was then inoculated in Liver Infusion Tryptose medium and J774 cells for the molecular identification and characterization of the parasite. The Pm strain of T. cruzi was identified by isolation in axenic culture medium, and based on the morphology, cell infection, growth kinetics, and molecular characterization. RESULTS: After isolation, the protozoan was identified as T. cruzi. No parasites were detected in the peripheral blood of the animal, which can be a characteristic inherent to the strain of T. cruzi that was isolated. Cell invasion assays were performed in triplicate in the J774 cell line to confirm the invasive ability of the Pm strain and revealed amastigote forms of the parasite within macrophages. CONCLUSIONS: Our biological and molecular characterizations helped understand parasite-host interactions and their evolutionary history in context of the associations between vectors, ecotopes, hosts, and groups of the parasite.

Extraction of Trypanosoma cruzi DNA from food: a contribution to the elucidation of acute Chagas disease outbreaks

Abstract: INTRODUCTION: Before 2004, the occurrence of acute Chagas disease (ACD) by oral transmission associated with food was scarcely known or investigated. Originally sporadic and circumstantial, ACD occurrences have now become frequent in the Amazon region, with recently related outbreaks spreading to several Brazilian states. These cases are associated with the consumption of açai juice by waste reservoir animals or insect vectors infected with Trypanosoma cruzi in endemic areas. Although guidelines for processing the fruit to minimize contamination through microorganisms and parasites exist, açai-based products must be assessed for quality, for which the demand for appropriate methodologies must be met. METHODS: Dilutions ranging from 5 to 1,000 T. cruzi CL Brener cells were mixed with 2mL of acai juice. Four Extraction of T. cruzi DNA methods were used on the fruit, and the cetyltrimethyl ammonium bromide (CTAB) method was selected according to JRC, 2005. RESULTS: DNA extraction by the CTAB method yielded satisfactory results with regard to purity and concentration for use in PCR. Overall, the methods employed proved that not only extraction efficiency but also high sensitivity in amplification was important. CONCLUSIONS: The method for T. cruzi detection in food is a powerful tool in the epidemiological investigation of outbreaks as it turns epidemiological evidence into supporting data that serve to confirm T. cruzi infection in the foods. It also facilitates food quality control and assessment of good manufacturing practices involving acai-based products.

Life cycle of Trypanosoma cruzi (y strain) in mice

Since 1958, we have studied experimental Chagas' disease (CD) by subcutaneous inoculation of 1,000 blood forms of Trypanosoma cruzi (Y strain) in Balb/C. mice. Evolution of parasitemia remained constant, beginning on the 5th and 6th day of the disease, increasing progressively, achieving a maximum on about the 30th day. After another month, only a few forms were present, and they disappeared from the circulation after the third month, as determined from direct examination of slides and the use of a Neubauer Counting Chamber. These events coincided with the appearance of amastigote nests in the tissues (especially the cardiac ones), starting the first week, and following the Gauss parasitemia curve, but they were not in parallel until the chronic stage. In 1997, we began to note the following changes: Parasites appeared in the circulation during the first week and disappeared starting on the 7th day, and there was a coincident absence of the amastigote nests in the tissues. A careful study verified that young forms in the evolutionary cycle of T. cruzi (epi + amastigotes) began to appear alongside the trypomastigotes in the circulation on the 5th and 7th post-inoculation day. At the same time, rounded, oval, and spindle shapes were seen circulating through the capillaries and sinusoids of the tissues, principally of the hematopoietic organs. Stasis occurs because the diameter of the circulating parasites is greater than the vessels, and this makes them more visible. Examination of the sternal bone marrow revealed young cells with elongated forms and others truncated in the shape of a "C" occupying the internal surface of the blood cells that had empty central portions (erythrocytes?). We hypothesize that there could be a loss of virulence or mutation of the Y strain of Trypanosoma cruzi.

Novo transmissor silvestre do "Trypanosoma (Schizotrypanum) cruzi" (Chagas, 1909)

In nests of opossum (Didelphis aurita), localized in palm-trees of the species Attalea indaya Dr., the authors found a new tritatoma, the description of which is being made by Dr. H. LENT. They verified that this triatoma had been naturally infected by Trypanosoma (Schizotrhypanum) cruzy. Two guinea-pigs were subsequently infected by peritoneal inoculation of excrements of this new triatoma. The xenodiagnosis of these guinea-pigs, made with normal nymphas of. T. megistus and T. infestans resulted positive after 25 days. Evidence was obtained of being the opossum (Didelphis) one of the sources infection of the new vector, because several specimens of them were found infected, and also a specimen of D. aurita, which contained trypanosomes with the morphology of T. (S.) cruzy in the peripheral blood.

Verificação de flagelados semelhantes ao Trypanosoma rangeli Tejera, em Rhodnius prolixus alimentados em caso de doença de Chagas na Venezuela: considerações sobre a natureza deste protozoário

1) - Por inoculação do sangue de um caso agúdo de moléstia de Chagas de Zaraza, Guárico, Venezuela, em cobaia (12-11-1940), obteve-se um Schizotrypanum com caractéres morfológicos distintos dos das amostras humanas comuns do Schizotrypanum cruzi; seu índice nuclear médio varia em tôrno de 1.4 e seu comprimento total médio é de 20 -21 μ (DIAS & FREITAS). 2) - Praticando-se, em 30-6-1941, no mesmo caso, o xenodiagnóstico com Rhodnius prolixus procedentes de criação normal de laboratório, verificou-se, nos barbeiros enviados para o Rio de Janeiro e dissecados 65 dias depois da sucção no paciente, a presença de flagelados com o aspecto do Trypanosoma rangeli Tejera, ao lado de tripanosomas metacíclicos semelhantes aos de Schizotrypanum. 3) Por inoculação do conteúdo intestinal dêstes Rhodnius em résus, isolou-se uma amostra de Schizotrypanum cujas formas sanguicolas tinham os mesmos caracteres da amostra obtida por inoculação do sangue do paciente em cobaia. 4) - No tubo digestivo de Rhodnius prolixus e de outros barbeiros criados no laboratório e infectados com a amostra de Schizotrypanum isolada por xenodiagnóstico, não foram observadas as formas carcterísticas do Trypanosoma rangeli, mas apenas crítidias e tripanosomas metacíclicos aparentemente indistinguíveis dos de amostras comuns de Schizotrypanum. 5) - Diante dos fatos referidos, sugere-se a hipótese de ser o Trypanosoma rangeli um Schizotrypanum patogênico do homem em seu ciclo no transmissor intermediário, alguns de cujos aspectos evolutivos são inconstantes e aparecem em circunstâncias indeterminadas.

Sôbre o Trypanosoma conorrhini, hemoparasito do rato transmitido pelo Triatoma rubrofasciata: presença do vector infectado na cidade do Rio de Janeiro

Os autores, após historiar a desccberta do Trypanosoma conorrhini (DONOVAN, 1909) (sinonimia Crithidia conorrhini DONOVAN, 1909, Trypanosoma boy lei LAFONT, 1912) no inseto transmissor e no hospedeiro vertebrado, o Rattus rattus diardi, fazem um estudo sumário do seu vector intermediário, o barbeiro cosmopolita Triatoma rubrofasciata (DE GEER, 1773) . Em seguida, referem a captura, no centro da cidade do Rio de Janeiro, Brasil, de um exemplar adulto dêste barbeiro parasitado por flagelados que foram identificados ao Trypanosoma conorrhini, fato êste pela primeira vez verificado no Novo Mundo. Certas formas evolutivas dêste parasito no inseto são muito semelhantes às do Schizotrypanum cruzi, mas os tripanosomas metacíclicos apresentam alguns caractéres morfológicos que permitem seu reconhecimento: os mais notáveis são o pequeno tamanho e a colocação do núcleo muito para trás, bem junto ao blefaroplasto. São dadas as medidas de 100 tripanosomas metaciclicos do T. conorrhini e de Schizotrypanum, cujo comprimento total médio foi, respectivamente, 13.88 u e 19.85 u. Nas seguintes espécies de barbeiro foi facilmente obtida a evolução do Trypanosoma conorrhini: Triatoma infestans, Triatoma vitticeps, Rhodnius prolixus e Panstrongylus megistus. Por inoculação de triatomas infectados foi obtida a transmissão do T. conorrhini ao camondongo branco, ao rato e ao macaco rhesus. As infecções foram muito fracas, custando-se a ver o tripanosoma no sangue, a fresco. Em gotas espêssas êle é encontrado com mais facilidade, mas o método de escôlha para verificação da sua presença no sangue dos animais é o xenodiagnóstico. Nesta prova o Triatoma infestans foi empregado com os resultados mais favoráveis. Por êste processo foi observada a infecção inaparente de um camondongo até 53 dias depois da inoculação. A forma sanguícola do T. conorrhini é um tripanosoma ttpico, de grande dimensões (40-60u), de extremidade posterior muito alongada e pontuda, membrana ondulante ampla e com largas pregas, blefaroplasto subcentral; em muitos indivíduos é notável uma estrutura particular, junto e à frente do blefaroplasto. Poucas horas depois de inoculados com as dejeções, os flagelados passam para a circulação, transformando-se em 3-4 dias nos grandes tripanosomas que acabam de ser descritos. O número de parasitos depende do número de flagelados inoculados. Não se conhece o processo de multiplicação do T. conorrhini no vertebrado, nunca tendo sido observados tripanosomas em via de divisão no sangue nem formas proliferativas nos tecidos. As sub-inoculações (de animal a animal) em geral não passam da primeira (MORISHITA, 1935). Tentativas até agora realizadas por outros pesquisadores no sentido de cultivar o T. conorrhini a partir do sangue de animais infectados, têm sido negativas. Por enquanto as pesquisas tendentes a determinar o hospedeiro vertebrado do parasito no Rio de Janeiro limitam-se a aplicação do xenodiagnós-tico em cinco ratos do Morro de Santo Antônio, cujo resultado...

Acerca del ciclo evolutivo del Trypanosoma (Schizotrypanum) cruzi Chagas 1909, en sus fases tisular y hematica

El autor pasa en revista los trabajos publicados sobre el ciclo evolutivo del Trypanosoma (S.) cruzi en el huésped vertebrado, desde el descubrimiento de la enfermedad hasta nuestros días. Luego analiza las ideas de los autores modernos, fundadas en gran parte en las observaciones que ya en 1914 realizaron MAYER y ROCHA LIMA de las cuales participan actualmente ROMAÑA y MEYER, ELKELES y WOOD. Finalmente expressa que a partir de los tripanosomas infectantes los parásitos que penetram en el protoplasma celular pueden seguir dos mecanismos en su evolución hacia cuerpos leishmanioides: 1.º) Por "regresión fusiforme" y 2.º) por "regresión orbicular"; llegados a la forma leishmanioide los parásitos se multiplican por división binaria, una vez lleno el protoplasma celular, siguen un processo inverso de transformación hacia tripanosoma que puede seguir igualmente dos mecanismos diversos: 1.) "progresión fusiforme" y 2.º) "progresión orbicular". Estos diversos mecanismos de transformación están esquematizados en la fig. N.º 1 del trabajo.