Rumen oligotrich protozoa, namely some species of
Entodinium, were successfully cultured
in vitro on a salt medium containing whole egg protein, proteose-peptone, rice starch, yeast extract, cellulose powder, and white clover juice. Of these constituents proteose-peptone, yeast extract, and cellulose powder could be omitted, but omission of the clover juice resulted in the death of the protozoa. Attempts to replace the clover juice by known compounds were unsuccessful.
Entodinia were maintained for over 10 months on a medium of rice starch and finely ground white clover at population densities of 2-10×10
4/ml. On the two media described above populations up to 2-3×10
5/ml were obtained by using a simple apparatus. On the latter medium, the ciliates were maintained at a mean density of nearly 2×10
5/ml for more than 50 days.
Entodiniumcaudatum, starting from a single cell, multiplied with the average division rate of approximately once per day during the initial 4 weeks. The clone was maintained for over 3 months with active multiplication. Morphological variation was observed in these clonal cultures.
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