A South African Babesia sp. of cattle which is as yet unclassified, was continuously cultivated in micro-aerophilous stationary-phase culture. The parasites were resuscitated from a blood stabilate stored in
liquid nitrogen. A modified HL-1 medium supplemented with either horse or bovine serum was used.
Cultures were initiated in a humidified atmosphere containing 2% 0₂ , 5% C0₂ and 93% N₂ at 37°C.
Parasites were detected on Giemsa-stained smears after 2 d in culture. On day 4, the cultures were
split at a ratio of 1:2 (v/v) and transferred into a humidified atmosphere of 5% C0₂ in air. Starting from
day 6, subcultures were made daily at a ratio of 1:4 (v/v). The percentage of parasitized erythrocytes
ranged from 2-5%. Addition of purine bases (hypoxanthine, adenine, adenosine or guanosine) was
essential for the continuous propagation of the parasites when bovine, but not horse serum, was used
for medium supplementation.
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