Growth of Babesia bigemina parasites in suspension cultures for vaccine production.

An Australian Babesia bigemina vaccine strain was maintained in suspension culture for 40 days. Parasite growth was compared using two tissue-culture flask sizes (25 and 75 cm2), four gas mixes (2%, 2.5%, 3% and 3.5% O2; 5% CO2; and the balance N2) and four packed blood cell (PCV) volumes (7%, 9%, 13% and 18%). The best continuous parasite yields were obtained from suspension cultures in 75-cm2 flasks at a PCV of 13% and gas mixtures of 2%-3% O2, 5% CO2 and the balance N2. Parasite yields per millilitre of culture medium were 3 times those obtained in microaerophilous stationary-phase cultures. The method has thus far been used for 6 months to produce the Australian requirements for live B. bigemina vaccine.