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Excystation and culture of Giardia spp. from human source
Abstract:
In this paper, we report the hatching of Giardia spp. trophozoites from purified cysts harvested in Sheather's solution from human feces. Isolated cysts were induced by acidic Hank's Balanced Salt Solution (HBSS), incubated at 37 degrees C for 30 minutes, concentrated by centrifugation and washed with pH 7.5 HBSS. The cysts were placed in 8 ml. culture medium (Pancreatic digest of casein and yeast extracts) supplemented with human serum and bile in borosilicate glass tubes with screw caps. Trophozoite excystation was scanned in a Commandon chamber filled with the suspension, sealed with a cover glass and vaseline. The trophozoite emergence was recorded with phase contrast in a photomicroscope. The excystation began within the first 3 minutes, after inoculation into medium. The trophozoites emerged trough a small hole at one end of the cyst and simultaneously began cellular division, the whole process required 5-30 minutes. The trophozoites attached themselves to the walls of the culture tube, permitting the replacement and removal of contaminants without dislodging them from the tube walls. The isolates MM-INP and MG-INP have been maintained at 37 degrees C for more than six months.
Insights
Researchers successfully hatched Giardia spp. trophozoites from purified cysts using acidic Hank
Area of Science:
- Medical Parasitology
- Microbiology
Background:
- Giardia spp. are significant enteric protozoan parasites.
- Excystation is a critical step for Giardia infection and propagation.
Purpose of the Study:
- To establish a reliable method for Giardia spp. trophozoite excystation from purified cysts.
- To observe and characterize the excystation process in vitro.
Main Methods:
- Purified Giardia cysts were induced using acidic Hank's Balanced Salt Solution (HBSS).
- Excystation was performed in a supplemented culture medium containing human serum and bile.
- Trophozoite emergence and subsequent development were monitored using phase contrast microscopy.
Main Results:
- Excystation initiated within 3 minutes of inoculation into the culture medium.
- Trophozoite emergence and cellular division occurred within 5-30 minutes.
- Developed trophozoites adhered to culture tube walls, facilitating maintenance and subculturing.
Conclusions:
- A reproducible method for Giardia spp. excystation was developed.
- The study provides insights into the early stages of Giardia trophozoite development.
- The established method allows for long-term maintenance of Giardia isolates for further research.