Zweygarth E, Van Niekerk C, Just M C, De Waal D T
Protozoology Division, Onderstepoort Veterinary Institute, South Africa.
Onderstepoort J Vet Res. 1995 Jun;62(2):139-42.
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% O2, 5% CO2 and 93% N2 at 37 degrees 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% CO2 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.
一种尚未分类的南非牛巴贝斯虫在微需氧固定相培养中连续培养。寄生虫从储存在液氮中的血液稳定物中复苏。使用添加了马或牛血清的改良HL-1培养基。培养在37℃、含2% O₂、5% CO₂和93% N₂的湿润气氛中开始。培养2天后在吉姆萨染色涂片上检测到寄生虫。第4天，将培养物按1:2（v/v）的比例传代并转移到含5% CO₂的空气湿润气氛中。从第6天开始，每天按1:4（v/v）的比例进行传代培养。被寄生红细胞的百分比在2%-5%之间。当使用牛血清而非马血清补充培养基时，添加嘌呤碱（次黄嘌呤、腺嘌呤、腺苷或鸟苷）对寄生虫的连续繁殖至关重要。
