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By Peter Malcolm Wallis, Brian R. Hammond
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Typical organelles characteristic for Giardia are seen within the cytoplasm, including a cyst wall, peritrophic space, flagellar axonemes, and curved portions of the adhesive disc, all of which are indistinguishable from those of control cysts formed in vivo. The cysts formed in vitro were round to oval in shape, ranging from 711 microns in width and 1114 microns in length, and had a cyst wall that, examined by SEM, appeared to be smooth. The striking ultrastructural similarity between cysts formed in vitro and viable Giardia cysts corroborated our prior assumption that the cysts formed in culture were indeed viable.
Figures 6 and 7. SEMs of a Giardia culture after exposure to bile for three days. Giardia cysts (arrowheads) formed in vitro were seen scattered between flagellated trophozoites, which were attached to the substratum. At higher magnification, seen in Figure 7, the contrast between the flagellated trophozoite and the in vitro formed cyst was readily apparent. The cysts were oval to round in shape and possessed a smooth surface, whereas the pearshaped trophozoites were flattened in a dorsalventral plane and attached to the substratum by their ventral adhesive disc.
A. Cysts formed in vitro were both morphologically and immunologically similar to Giardia cysts formed in vivo and also were viable, as demonstrated by the uptake of fluorogenic dyes. The inability of Giardia trophozoites to undergo encystation in vitro has raised questions as to whether host related factors, such as gutassociated microorganisms or nutritive factors and stimuli from the small intestine, might play a role in this process. This model would be beneficial in many aspects of Giardia research, including biochemical analysis of the cyst and the cyst wall, development of giardicidal agents involved in blocking cyst wall formation, the testing of Kochs' postulates in regard to infectivity of cysts, and the investigation of questions related to nuclear division and cyst wall production.
Advances in Giardia Research by Peter Malcolm Wallis, Brian R. Hammond