Multiple loop purification method for selective cultivation of Pentatrichomonas hominis.
It is always troublesome having protozoan cultures contaminated with other organisms in the laboratory. The method described here produces high efficiencies of purification for fast moving flagellate protozoa. A human strain Pentatrichomonas hominis was employed in the study to examine the effects of multiple loop tubes on the purification of flagellates. Trichomonads were harvested from a trypticase yeast extract iron-serum-33 (TYI-S-33) medium, adjusted to 2 X 10(5) organisms/ml, and mixed with an equal volume of 2 X 10(6) organisms/ml of bacteria. The isolation was performed at 37 degrees C in TYI-S-33 medium containing a suitable amount of antibiotics (1000 U/ml of penicillin, 1000 micrograms/ml of streptomycin, and 4 micrograms/ml of fungizone). Four days later, 10(6) organisms/ml of protozoa, free of bacteria, were observed at the other end of the single loop and the double loop tubes. About the same amount of flagellates could be found at the other end of the triple loop tube six days after incubation. The traditional U-shaped tubes were used as controls and 10(5) cells/ml of flagellates were recovered in the presence of bacteria two days after incubation. An axenic culture of P. hominis was successfully isolated from the feces of a Formosan rock-monkey, Macaca cyclopsis, by this method. Purified trichomonads were recovered from a double loop purification tube five days after incubation.
Chao
D
D
Huang
B C
BC
Chen
Y A
YA
Wang
S R
SR
D000818:Animals; D003584:Cytological Techniques / Q000295:instrumentation; D056890:Eukaryota / Q000254:growth & development* / Q000302:isolation & purification; D005243:Feces / Q000469:parasitology; D006801:Humans; D008251:Macaca