SYNOPSIS. A method for the in vitro culture of O. purkynei was developed which permitted the continuous culture of this protozoon in clone culture for a. period of 32 months. The shortest division time was 24 hr. The concentration of cells varied between 700 and 1000/ml in the routine procedure. Variations in spination which occurred in the clone culture suggested that this characteristic was of doubtful taxonomic importance. Ground wheat and alfalfa served as substrates but soluble sugars did not. Green plant material appeared to be necessary for continued growth of the protozoa. Ingestion of a large streptococcus was demonstrated by vital staining of a mixed population of bacteria with tetrazolium prior to incubation with the protozoan suspension in the presence of wheat. O. purkynei can tolerate exposure to variations in osmotic pressure, temperature, and oxygen which are consonant with its transfer in nature by grooming or ingestion of contaminated food or drink.
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Robert A. Mah (1964) studied this question.