By Peter Malcolm Wallis, Brian R. Hammond
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By day three, these clusters had increased in size and included trophozoites as well as cysts. The other type of cyst (not shown) was of similar size, shape, and had a cyst wall, but appeared nonviable since the cyst usually lacked the presence of typical organelles or contained two to four nuclei within a shrunken cytoplasmic mass. A wide variation in cyst viability was detected using the incorporation of FDA, ranging from 4050% to less than 1%, in Giardia cysts formed in vitro. 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.
Lamblia extracts apparently had no deleterious effects and did not induce any morphological changes compared with TPS1 and MEM control culture cells in either HeLa or Vero cells till 96 h after interaction. lamblia are shown in Figure 3 (AE). The first interaction was the adhesion of trophozoites to the cell surface followed by progressive destructive changes in the cell. The lateral shield along the periphery of the trophozoites and the ventrolateral flange could be seen in direct contact with both the cell surface in some and the coverglass in others.
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.