巴尔通体液体培养条件简化及生长曲线观察

Optimization of liquid growth conditions and determination of growth curves for Bartonella species

摘要:

【目的】应用一种昆虫细胞培养基作为基础成分培养巴尔通体(Bartonella species), 建立一种操作方便、高效稳定的巴尔通体液体培养方法。【方法】昆虫细胞培养基中添加10%胎牛血清, 以此为基础培养液分别添加蔗糖和谷氨酰胺, 比较这两种成分对汉赛巴尔通体(B. henselae)和五日热巴尔通体(B. quintana)生长的影响并观察其他10种巴尔通体在简化后的培养液中的生长特性。【结果】添加蔗糖和谷氨酰胺不会明显促进巴尔通体的生长, 10种巴尔通体在简化后的培养液中均生长良好。不同种巴尔通体生长曲线不同, 汉赛巴尔通体和五日热巴尔通体的世代时间分别为5.2 h和4.3 h, 生长速度快于固体培养。【结论】以昆虫细胞培养基作为基础成分的培养液适于巴尔通体液体培养, 特别是对一些更难培养的巴尔通体提供了一种较好的培养方法。

Abstract:

[Objective] We tested an insect cells-based liquid medium and its utility for the easy-to-maintain, fast and reliable system for cultivation of different Bartonella. [Methods] Media composition and growth conditions were optimized using B. henselae and B. quintana. The growth curves of the other ten Bartonella species were determined using the optimized liquid medium. [Results] Glutamine and sucrose supplements did not affect significantly the growth rate of these bacteria. A generation time of 5.2 h for B. henselae and 4.3 h for B. quintana was estimated during the exponential phase of the growth curve occurred between days 1 and 2 after inoculation. The twelve medical and veterinary important Bartonella species grew well under conditions established and each exhibited unique growth characteristics. [Conclusion] This liquid growth medium may provide an advantage over conventional direct blood agar plating for analyzing Bartonella properties including antibiotic susceptibility and resistance and pathogenicity.