Rapid isolation of metacyclic trypomastigotes of Trypanosoma cruzi from feces and urine of the vector.
Metacyclic trypomastigotes of Trypanosoma cruzi from infected Triatoma infestans were isolated and purified by an improved procedure. By using feces and urine of bugs the duration and the number of steps for purification were greatly reduced, thereby minimizing any possible effects on the flagellates. After DEAE-Sephacel column chromatography, a buffer supplemented with bovine serum albumin protected against the slight surface alterations of the trypomastigotes otherwise observed. About 10(7) metacyclic trypomastigotes (including 30% intermediate flagellates) were obtained from 100 bugs. The intermediate forms possessed the surface charge of trypomastigotes, and the kinetoplast was beside or posterior to the nucleus but not subterminal. Further increases in the number of bugs and the purified trypomastigotes were also possible with only slight increases in the time needed for isolation. In a comparison of complement-mediated lysis resistance and infectivity for mice, pre- and post-separation trypomastigotes showed no significant differences.
Institut für Biologie I (Zoologie), Albert-Ludwigs-Universität, Freiburg, F.R.G.
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Schaub
D000818:Animals; D014355:Chagas Disease / Q000469:parasitology*; D002850:Chromatography, Gel; D005243:Feces / Q000469:parasitology; D005898:Glass; D007303:Insect Vectors / Q000469:parasitology*; D007814:Larva / Q000469:parasitology; D051379:Mice; D000267:Tissue Adhesions; D014224:Triatoma / Q000469:parasitology*; D014349:Trypanosoma cruzi / Q000302:isolation & purification*; D014556:Urine / Q000469:parasitology