Blastocystis hominis: pathogenic potential in human patients and in gnotobiotes.

Germfree guinea pigs were inoculated orally, in some experiments, and intracecally, in others, with Blastocystis hominis and the enteric flora from symptomatic patients. Other germfree guinea pigs received the parasite from axenic culture and still others from monoxenic culture with Proteus vulgaris. Fourteen of 43 animals inoculated orally with B. hominis and patient's enteric flora developed B. hominis infections and those with particularly heavy infections developed watery diarrhea of more than 1 week's duration immediately prior to sacrifice. Similar results were obtained from intracecal inoculations in that 13 of 28 animals developed infections and those with the greatest numbers of B. hominis had watery diarrhea for more than 1 week prior to sacrifice. Gross pathologic changes in these animals were mostly unremarkable, with only a slight hyperemia observed in several of the symptomatic animals. Microscopic examination, however, revealed frequent penetration of intestinal epithelium by B. hominis and the parasites in significant numbers were observed within the epithelium. There was a slight increase in cellularity in the lamina propria but parasites were not observed therein and their presence in the epithelium did not provoke an inflammatory response. Only one of eight animals inoculated from monoxenic cultures developed B. hominis infection (asymptomatic), and infections were not produced in animals inoculated from axenic culture. As a result of our observations of diarrhea in patients with particularly heavy infections with B. hominis together with the demonstration of similar symptoms in animals heavily infected with this parasite, we believe B. hominis may occasionally be related causally to the production of such symptoms by a mechanism not completely understandable.     

Entamoeba histolytica and Giardia lamblia: effects of cysteine and oxygen tension on trophozoite attachment to glass and survival in culture media

Attachment of Entamoeba histolytica and of Giardia lamblia trophozoites to glass was monitored during the culture cycle. Attachment of each parasite was greatest during the exponential phase of axenic growth. The effects of l-cysteine upon the kinetics of attachment of trophozoites to glass were determined quantitatively. Attachment in complex growth media required cysteine, even under N₂, atmosphere. With cysteine, the rates of attachment were greatest for the first 2 hr, then continued more slowly. The numbers of attached trophozoites decreased immediately upon exposure to medium without cysteine. The role of cysteine in protecting trophozoites of both species from the lethal effects of oxygen was assessed using clonal growth in agar or agarose medium to determine viability following exposure to varying oxygen tensions in liquid medium. Cysteine was required for viability of trophozoites. Without cysteine, decreasing the oxygen tension prolonged survival. Under increased oxygen tension, cysteine delayed the onset of exponential killing. Although it has no thiol reducing group, l-cystine similarly protected E. histolytica.     

Trypanosoma brucei: morphometric changes and loss of infectivity during transformation of bloodstream forms to procyclic culture forms in vitro.

The transformation of Trypanosoma brucei bloodstream forms to procyclic culture forms in the semidefined medium SDM-77 has been studied by light microscopy and quantitative electron microscopy. Stumpy and intermediate forms are able to transform to culture forms whereas slender forms die after approximately 24 hr. The surface coat and infectivity for the mammalian host are lost after 72 hr. Morphometrical analysis of the cells during transformation process revealed: (1) The cytoplasm and the cell surface increased significantly; (2) the volume density of the mitochondrion increased twofold and the surface density of the inner mitochondrial membrane increased threefold; (3) the volume density of the glycosomes remained about constant; (4) the volume density of the lipid inclusions increased up to 72 hr, probably as a result of the complete oxidation of glucose. Transformation as observed by light microscopy was completed in 72 hr. However, quantitative electron microscopy revealed that establishment of the culture form was incomplete even after 11 days.