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Multiple loop purification method for selective cultivation of Pentatrichomonas hominis
Proceedings of the National Science Council, Republic of China. Part B, Life Sciences
|January 1, 1987
Summary
Contaminated protozoan cultures are a common lab problem. This study presents a novel multiple loop tube method for efficiently purifying fast-moving flagellates like Pentatrichomonas hominis, achieving bacteria-free cultures.
Area of Science:
- Microbiology
- Parasitology
- Cell Biology
Background:
- Protozoan cultures are frequently contaminated with bacteria and fungi in laboratory settings.
- Maintaining pure protozoan cultures is essential for accurate research and experimentation.
- Fast-moving flagellates, such as Pentatrichomonas hominis, pose unique challenges for purification due to their motility.
Purpose of the Study:
- To evaluate the efficacy of a novel multiple loop tube method for purifying fast-moving flagellate protozoa.
- To compare the purification efficiency of single, double, and triple loop tubes against traditional U-shaped tubes.
- To establish an axenic culture of Pentatrichomonas hominis from a non-human primate source.
Main Methods:
- Flagellate protozoa (Pentatrichomonas hominis) were intentionally contaminated with bacteria.
- Cultures were incubated in trypticase yeast extract iron-serum-33 (TYI-S-33) medium with antibiotics at 37°C.
- Purification was assessed using single, double, and triple loop tubes, with traditional U-shaped tubes serving as controls.
Main Results:
- Bacteria-free flagellates were recovered at high concentrations (10^6 organisms/ml) in double loop tubes within four days.
- Triple loop tubes yielded similar results after six days, while single loop tubes also showed high purification efficiency.
- Traditional U-shaped tubes showed lower recovery rates (10^5 cells/ml) and persistent bacterial contamination.
- An axenic culture of Pentatrichomonas hominis was successfully isolated from monkey feces using the double loop method.
Conclusions:
- The multiple loop tube method offers a highly efficient and effective approach for purifying fast-moving flagellate protozoa.
- This technique significantly reduces contamination issues, enabling the establishment of pure protozoan cultures.
- The successful isolation of an axenic culture from a primate source demonstrates the broad applicability of this purification method.