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.

Haematophagic behavior in laboratory of Lutzomyia cruzi (Mangabeira) (Diptera: Psychodidae) in relation to three mammalian blood sources in Manaus, Brazil

The sand fly Lutzomyia cruzi is considered as one of vectors of visceral leishmaniasis in Brazil. This work examined optimum feeding age, feeding time, host preference, fecundity rates, and female blood meal volume taken by single females from a closed colony of L. cruzi. Mean feeding time was longer on hamsters, 6.6 minutes, than on humans, 5.7 minutes. 49.1% of the 48h-old flies fed on humans and 43.3% of 72h-old flies fed on hamsters. Of a total of 120 females, 61% fed on humans and 25% fed on hamsters. Total fecundity was significantly higher in females fed on hamster than on human or opossum. Laboratory-reared L. cruzi females fed earlier, more promptly, and preferably on humans than on hamsters when offered these blood-meal sources simultaneously. The blood-meal volume is higher in females fed on hamsters than other hosts (human and opossum).

Nova tripanozomiaze humana: estudos sobre a morfolojia e o ciclo evolutivo do Schizotrypanum cruzi n. gen., n. sp., ajente etiolojico de nova entidade morbida do homem

1º O Schizotrypanum cruzi apresenta, no organismo do conorrino, duas modalidades de desenvolvimento, reprezentando a primeira simples cultura do parazito; a outra, provavelmente precedida de fenomenos sexuais não surpreendidos, será, talvez, o ciclo evolutivo, eficaz na transmissão entre os vertebrados. 2º O Conorhinus é um verdadeiro hospede intermediario do Schizotrypanum, cujo ciclo é realizado num prazo minimo de 8 dias. 3º Os flajelados, com tipo de critidias encontrados nos conorrinos em liberdade, podem reprezentar estádios culturais do Schizotrypanum ou serão parazitos excluzivos do inséto. 4º A ocurrencia do ciclo evolutivo sexuado, no organismo do conorrino, depende de condição não explicada, dos flajelados no sangue dos vertebrados. Ao terminar, cumprimos o grato dever de afirmar o maior reconhecimento ao nosso mestre e Diretor Dr. Gonçalves Cruz, a cuja orientação devemos o rezultado destas pesquizas. Somos ainda profundamente grato aos nosso mestres, Professores S. von PROWAZEK e M. HARTMANN, de quem recebemos os melhores ensinamentos para concluzão deste trabalho. Tambem somos em extremo obrigado ao Dr. ADOLPHO LUTZ, cujo auxilio nos foi do mais alto proveito. Tivemos sempre, como esforçado companheiro de trabalho na zona infestada pela nova especie morbida, o Dr. BELISARIO PENNA, a quem devemos os inestimaveis proveitos de um auxilio eficaz.

O gênero Schizotrypanum Chagas, 1909

No presente trabalho, em que foram analisados os caractéres de Schizotrypanum e consideradas suas relações com os de outros flagelados digenéticos, acreditamos ter ficado bem demonstrado que este gênero encontra sólidos fundamentos em que se baseie. Schizotrypanum possue caractéres morfológicos peculiares, que o aproximam de Leishmania no periodo de multiplicação e de Trypanosoma na fase sanguinea. Os flagelados pertencentes a esse gênero caracterisam-se não só pela morfologia da fórma de tripanosoma, como pela evolução no organismo do vertebrado. No S. cruzi, como no S. vespertilionis, a multiplicação se processa nos tecidos, constando da divisão binaria das formas intracelulares de leishmania; os tripanosomas sanguicolas não se multiplicam. Nenhum Trypanosoma apresenta no mamífero uma evolução morfológicamente e ecológicamente identica á de Schizotrypanum. S. cruzi aproxima-se dos tripanosomas patogenicos pela morfologia da fórma sanguicola e pela virulencia ás vezes mortal para o homem e diversos animais; deles se afasta entretanto pela evolução no inséto, modo de transmissão e facil cultivabilidade, caractéres biológicos estes que são comuns aos tripanosomas não patogenicos. Em nenhum dos grupos de tripanosomas póde o S. cruzi ser rigorosamente incluido, deles se distinguindo facilmente seja por sua morfologia, seja por sua biologia. O conjunto de caractéres próprios fundamenta perfeitamente a manutenção do gênero de Chagas, indicando-lhe como situação mais adequada, na classificação dos tripanosomídeos de mamíferos, o logar intermediario entre Leishmania e Trypanosoma. A separação do gênero Schizotrypanum é o melhor caso, quiça o unico justificado, dentre as numerosas tentativas para o desmembramento de Trypanosoma. Ela se impõe como medida compreensiva e util para a coordenação dos membros da complexa familia dos tripanosomideos e se justifica á luz dos mais exigentes critérios sistematicos.