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Two strains of Trypanosoma brucei were propagated from the salivary glands of 5 Glossina morsitans for more than 200 days on a bovine embryonic spleen feeder layer using buffered RPMI 1640 medium supplemented with 20% bovine fetal serum. In the first 2 to 3 weeks of cultivation the density of parasites in the salivary glands and culture medium remained constant probably because of defective binary fission. The parasites were infective to rodents only on days 17 and 25. Electron microscopic examination of the parasites on 6 different occasions revealed that they were similar to the immature metatrypomastigotes of T. brucei described in the salivary glands of infected tsetse flies.
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