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Axenic Culture Studies of Entamoeba Species

Axenic Entamoeba invadens could be grown at room temperatures in the diphasic types of medium used for E. histolytica. Conversely, axenic E. histolytica from diphasic cultures could at ca. 37°C, within limits, be adapted into variations of the monophasic media originally developed for E. invadens. Only occasional E. histolytica inocula grew for more than one or two weekly subcultures in such monophasic media, but several series have been transferred up to seven times.

Of the various natural ingredients in E. histolytica media, only serum was required at consistently high levels (ca. 10%). E. invadens has been maintained for several weeks in chemically defined and partially defined media, but population increases were not observed. The survival of E. invadens but not E. histolytica was significantly extended by helping to preserve anaerobic conditions in cultures with a petroleum jelly seal.

Zone electrophoresis in starch gel combined with chemically specific staining for enzymatic activity was used in an attempt to compare the esterase enzymes of the two axenically cultured species. However, only esterases associated with the culture media and not with the amebae themselves were demonstrated.