A criptosporidiose humana é uma infecção causada pelo Cryptosporidium sp, um protozoário coccídio de patogenicidade emergente e responsável por severa e prostrante diarréia aquosa em humanos, principalmente em indivíduos imunodeprimidos. O diagnóstico, feito através da utilização de esfregaços de fezes submetidos a técnicas de concentração e coloração específica pela fucsina-carbólica tem oferecido bons resultados em nosso laboratório. Tendo em vista o longo tempo despendido para a observação dos esfregaços considerando-se a pequenez das formas e o contraste da coloração, realizamos modificações no procedimento técnico da coloração ácido-resistente que resultaram em sensível melhoria das preparações: a fucsina-carbólica passou a ser deixada sobre o esfregaço por período de 3 minutos (LENNETTE et al., 1985) e procedeu-se a substituição da solução de álcool-ácido sulfúrico a 5% (HENRIKSEN & POHLENZ, 1981) por solução de ácido clorídrico a 0,5% em álcool etílico 70%, por cerca de 2 minutos (contribuição original). Estas alterações promoveram melhor remoção do excesso de fucsina-carbólica, aumentando a eficiência da etapa de descoloração e consequentemente otimizando o contraste do processo de coloração. Nestas condições, as lâminas examinadas em microscópio óptico em aumentos de 250x e 1000x tiveram a visualização dos oocistos do protozoário facilitada, sendo os mesmos observados em contraste destacado com pigmentação intensa de cor rosa-avermelhada contra coloração de fundo azulada. Vale destacar que estas modificações oferecem vantagens de rápido processamento do material e facilidade de visualização do protozoário, diminuindo o tempo de microscopia, tornando a análise das lâminas mais rápida e menos cansativa, agilizando o diagnóstico.

Human cryptosporidiosis is an infection caused by Cryptosporidium spp, a coccidial protozoan of emergencial pathogenicity and responsible for severe and prostrating watery diarrhea, mainly in immunocompromised patients. Smears of stools submitted to concentration and staining by carbol fuchsin technique has been used in our laboratory as a diagnostic procedure for cryptosporidiosis. The long time observing the smears in the microscope due to the small size of the forms and the low contrast of the staining led us to introduce some modifications in the original protocol for the acid-fast staining. The smears were treated with the carbol fuchsin solution for 3 minutes as recommended by LENNETTE et al., 1985 and the solution of the ethyl alcohol 70%-chloride acid 0.5% was used instead of the ethyl alcohol-sulfuric acid 5% recommended by HENRIKSEN & POHLENZ, 1981. Smears were treated with the discoloration solution for 2 minutes. These modifications promoted a better washing out of the excess of carbol fuchsin therefore increasing the dye efficiency. In such conditions, the visualization of protozoan oocysts on the slides examined became easier. They were observed by the contrast of their intense pink-reddish pigmentation against the blue background. It's worthwhile to emphasize that these modifications offer advantages when time and accuracy are concerned.

The aim of this paper is to present the analysis of sexual morphological differences observed in 5th instar nymphs of the following species: Panstrongylus megistus; Rhodnius neglectus; Triatoma brasiliensis; T. infestans; T. matogrosensis and T. tibiamaculata. Male and female nymphs were examined and photographed with a Scanning Electron Microscope. The 9th segment dimensions of dorsal and ventral faces were determined through a Profile Projector. Results and statistical analysis showed significant differences: the 9th sternite is significantly broader in male than female nymphs, while in five species; tergites in female nymphs are broad and in male are narrow.

Camundongos normais, utilizados como filtros biológicos, foram inoculados endovenosamente, com tripomastigotas sangüícolas da derivada RCL da cepa RC do T. cruzi. Decorridas 48 horas, os animais foram submetidos ao xenodiagnóstico e à punção cardíaca para semeadura do sangue em meio de Warren. No estômago dos triatomíneos e em cultura a 28°C, os tripomastigotas diferenciaram-se em formas arredondadas (esferomastigotas e/ou amastigotas). Esse comportamento das formas sangüícolas largas tem sido observado com freqüência e nos leva a inferir que a biologia do T. cruzi não estaria apenas relacionada com a cepa, mas, eventualmente, com populações do parasita.

The study of the behaviour of T. cruzi strains in animals as in human beings, runs into great difficulties when the attempt is made to relate the morphology of this parasite to its biological behaviour. The thin forms are able to penetrate vertebrate cells faster than the broad forms. So, faced with the biological complexity of the T. cruzi strains and substrains it was decided to investigate the behaviour of its blood forms obtained from normal mice as biological filters. These mice were endovenously inoculated with blood trypomastigotes from derivated RCL of the T. cruzi RC strain. Forty-eight hours later, the animals went through xenodiagnosis and heart punch in order to sow the blood in Warren's medium. The trypomastigotes became round (spheromastigotes and/or amastigotes) in the stomach of the triatomines and cultures at 28°C. The behaviour of the broad forms has been frequently observed and leads to the conclusion that the biology of the T. cruzi is not only related to the strain, but, sometimes, to the parasite population.

Foram utilizados camundongos brancos, pesando em média 18g e duas cepas de Trypanosoma cruzi, morfologicamente distintas: Y com predominância de formas sangüíneas delgadas e Bolívia com predomínio de formas largas. Os lotes de animais receberam 2 x 10³, 2 x 10(4) e 2 x 10(5) tripanossomos por animal e as vias de inoculação utilizadas foram a intraperitoneal e a subcutânea. Nos animais foi observado o curso de infecção. Os resultados obtidos revelaram que, nos experimentos em que se utilizou a cepa Y, existem algumas diferenças significantes, com infecções mais uniformes e virulentas, após inoculação subcutânea de 2 x 10³ e 2 x 10(4) formas sangüíneas de T. cruzi. Entretanto, isto não ocorreu com a cepa Bolívia, pois os animais apresentaram o mesmo padrão de parasitemia e os demais caracteres morfológicos, quer se utilizasse a via subcutânea ou intraperitoneal. Tal fato permite sugerir a existência de interrelação entre os fatores via de inoculação traduzida pela maior ou menor presença de macrófagos no sítio de inoculação, e a morfologia das formas sangüíneas representada pela maior ou menor capacidade de penetração celular.

By means of comparative studies of the infection of mice with T. cruzi using intraperitoneal and subcutaneous inoculation, it was concluded that more uniform and virulent infections are obtained after subcutaneous inoculation. This is due to an immediate cellular response, when intraperitoneal inoculations are used. With the aim of making comparative studies as between the two methods of inoculation, the intraperitoneal using two, morfologically distinct, strains of T. cruzi: Y with predominance of thin forms and Bolivia with predominance of broad forms. White mice of approximately 18g were required for these tests. The groups of animals received 2 x 10³, 2 x 10(4) and 2 x 10(5) trypanossomes per animal, and the course of the infection was subsequently observed. The results revealed that after subcutaneous inoculation when the Y strain was used in the tests, some significant differences appear, with more virulent and uniform infections. However, these did not appear when the Bolivia strain was used, because the animals showed the same standard parasitemy and other morpho-biological features, whether by subcutaneous or intraperitoneal inoculation. This incident suggests the existence of interrelation between the factors: method of inoculation represented by a greater or lesser presence of macrophages in the inoculated area, and the morphology of the blood forms represented by a greater or lesser capacity for cellular penetration.

Foram utilizados camundongos brancos, pesando em média 18g e duas cepas de Trypanosoma cruzi, morfologicamente distintas: Y com predominância de formas sangüíneas delgadas e Bolívia com predomínio de formas largas. Os lotes de animais receberam 2 x 10³, 2 x 10(4) e 2 x 10(5) tripanossomos por animal e as vias de inoculação utilizadas foram a intraperitoneal e a subcutânea. Nos animais foi observado o curso de infecção. Os resultados obtidos revelaram que, nos experimentos em que se utilizou a cepa Y, existem algumas diferenças significantes, com infecções mais uniformes e virulentas, após inoculação subcutânea de 2 x 10³ e 2 x 10(4) formas sangüíneas de T. cruzi. Entretanto, isto não ocorreu com a cepa Bolívia, pois os animais apresentaram o mesmo padrão de parasitemia e os demais caracteres morfológicos, quer se utilizasse a via subcutânea ou intraperitoneal. Tal fato permite sugerir a existência de interrelação entre os fatores via de inoculação traduzida pela maior ou menor presença de macrófagos no sítio de inoculação, e a morfologia das formas sangüíneas representada pela maior ou menor capacidade de penetração celular.

By means of comparative studies of the infection of mice with T. cruzi using intraperitoneal and subcutaneous inoculation, it was concluded that more uniform and virulent infections are obtained after subcutaneous inoculation. This is due to an immediate cellular response, when intraperitoneal inoculations are used. With the aim of making comparative studies as between the two methods of inoculation, the intraperitoneal using two, morfologically distinct, strains of T. cruzi: Y with predominance of thin forms and Bolivia with predominance of broad forms. White mice of approximately 18g were required for these tests. The groups of animals received 2 x 10³, 2 x 10(4) and 2 x 10(5) trypanossomes per animal, and the course of the infection was subsequently observed. The results revealed that after subcutaneous inoculation when the Y strain was used in the tests, some significant differences appear, with more virulent and uniform infections. However, these did not appear when the Bolivia strain was used, because the animals showed the same standard parasitemy and other morpho-biological features, whether by subcutaneous or intraperitoneal inoculation. This incident suggests the existence of interrelation between the factors: method of inoculation represented by a greater or lesser presence of macrophages in the inoculated area, and the morphology of the blood forms represented by a greater or lesser capacity for cellular penetration.

Camundongos normais, utilizados como filtros biológicos, foram inoculados endovenosamente, com tripomastigotas sangüícolas da derivada RCL da cepa RC do T. cruzi. Decorridas 48 horas, os animais foram submetidos ao xenodiagnóstico e à punção cardíaca para semeadura do sangue em meio de Warren. No estômago dos triatomíneos e em cultura a 28°C, os tripomastigotas diferenciaram-se em formas arredondadas (esferomastigotas e/ou amastigotas). Esse comportamento das formas sangüícolas largas tem sido observado com freqüência e nos leva a inferir que a biologia do T. cruzi não estaria apenas relacionada com a cepa, mas, eventualmente, com populações do parasita.

The study of the behaviour of T. cruzi strains in animals as in human beings, runs into great difficulties when the attempt is made to relate the morphology of this parasite to its biological behaviour. The thin forms are able to penetrate vertebrate cells faster than the broad forms. So, faced with the biological complexity of the T. cruzi strains and substrains it was decided to investigate the behaviour of its blood forms obtained from normal mice as biological filters. These mice were endovenously inoculated with blood trypomastigotes from derivated RCL of the T. cruzi RC strain. Forty-eight hours later, the animals went through xenodiagnosis and heart punch in order to sow the blood in Warren's medium. The trypomastigotes became round (spheromastigotes and/or amastigotes) in the stomach of the triatomines and cultures at 28°C. The behaviour of the broad forms has been frequently observed and leads to the conclusion that the biology of the T. cruzi is not only related to the strain, but, sometimes, to the parasite population.

Os autores realizaram um estudo do comportamento da infecção determinada por uma amostra do Trypanosoma cruzi - amostra Bolivia - através da inoculação de formas aflageladas ou flageladas, obtidas de cultura em meio de' Warren, em camundongos brancos jovens. Utilizaram como critérios de avaliação do decurso da infecção: o período prepatente, o índice de infecção, a parasi temia, a duração da fase aguda e a taxa de letalidade. Essas observações indicaram que o comportamento da infecção varia quando partimos de um ou outro tipo de inóculo.

The authors made a study of the infection behaviour of a T. cruzi strain - Bolivia strain - through the inoculation of leishmania and flagellates forms from culture (Warren media) in white baby mice. They have used the prepatent period, the infection index, the parasitemia during the acute phase and the death rate for the evaluation of the infectious process. The results showed different characteristics of the infection when are used one or the other kind of inoculation.