Studies on Entamoeba histolytica; the relation of oxidation-reduction potentials to the growth, encystation and excystation of Entamoeba histolytica in culture.

1. The oxidation-reduction potential in cultures of E. histolytica has been studied. It was found that a close relation exists between the potential in the culture and the growth, encystation, and excystation of E. histolytica. The small differences observed in different cultures indicate the reproducibility of the potentials and the reliability of the potential condition in explaining these biological activities of the amoebae in culture. 2. The best growth of trophozoites was observed at potentials between −350 and −425mV., less at potentials between −275 and −350mV., still less at between −200 and −275mV. At potentials between −150 and −200mV. a bare increase in number of trophozoites was noticed. Between potentials of −50 and −150mV. no increase of growth was observed. Trophozoites were found dead in 12 hr. or more at potentials between −50 and +50mV., and died and disintegrated in a couple of hours between potentials of +100 and +200mV. 3. The life span of a culture of E. histolytica was also found to be related to the potential condition. The longer the potential in the culture remains below −150mV., the longer the culture remains positive. 4. Excystation of E. histolytica, no matter in what media, took place only at potentials of −115mV. or lower. 5. Mass encystation of E. histolytica was induced in cultures in which a ‘sharp rise of potential’ occurred after a greatly increased rate of multiplication of trophozoites after 22–24 hr. of incubation. Maturation of cysts occurred during this period of ‘sharp rise of potentials’. If the potential went down shortly after its sharp rise, the cysts failed to become mature and disappeared in about 12 hr. 6. Trophozoites and cysts observed in cultures with very low potentials (below −300mV.) and a rapid multiplication were definitely smaller than those in cultures having higher potentials and trophozoites multiplied slowly. 7. In cultures of amoebae in which mass encystation failed to occur, or mass encystation occurred but cysts failed to become mature, some success was observed in a few trials by artificially creating the ‘sharp rise of potential’ by oxygenation. 8. The pH values noticed in cultures at times of encystation ranged from 6.4 to 6.8. This pH range was a little lower than that observed previously. Mass encystation failed to occur at the change in potential if the pH was 7.2 or higher. 9. It is felt that the process of encystation requires, at least, two extrinsic and one intrinsic factors. The first two are the potential change from more favourable to less favourable conditions and the pH. The last one is the tendency to encyst possessed by the amoebae. 10. The low potential condition in a culture of E. histolytica was found chiefly provided by the activity of the accompanying bacteria. It is, therefore, felt that the indispensability of a bacterial flora for cultivation of amoebae may be explained, partly if not entirely, by the potential relation. To verify this statement, it is hoped to propagate E. histolytica in cultures in which a strongly reducing condition is provided without the presence of bacteria. The author wishes to acknowledge the kindness of Dr David Weinman of this department in making many invaluable corrections of this manuscript, and he is very grateful to Mrs Elizabeth B. Keller and Mrs Wanda Reed for their help in preparing the graphs accompanying it.