Institut für Biologie, Molekulare Ökologie, Martin-Luther-Universität Halle-Wittenberg, Hoher Weg 4, 06099 Halle, Saale, Germany.
J Eukaryot Microbiol. 2011 Jan-Feb;58(1):7-10. doi: 10.1111/j.1550-7408.2010.00514.x. Epub 2010 Dec 3.
The protozoan parasite Crithidia bombi and its host, the bumblebee Bombus terrestris, are used as a model system for the study of the evolutionary ecology of host-parasite interactions. In order to study these interactions we established a method for in vitro cultivation of single parasite strains. Additionally, a high-throughput method is developed for the determination of cell numbers in cultures by means of optical density (OD) measurements. The protocol for in vitro cultivation allowed for growing different strains on agar plates as well as in culture medium. A calibration curve for the relationship between cell number and OD has been developed. Subsequently, growth rates for different genotypes of C. bombi have been recorded. Significant differences in the growth rates and generation times between these genotypes were demonstrated. As this might be related to the virulence of the parasite, this relationship may be confirmed by in vivo growth rate determination. In comparison with conventional cell counting, the application of OD measurements allows for high-throughput experiments as the time taken to record each sample is reduced by a factor of 30. The in vitro cultivation method allows for controlled infection experiments in order to study host-parasite interactions.
原生动物寄生虫 Crithidia bombi 及其宿主熊蜂 Bombus terrestris 被用作研究宿主-寄生虫相互作用进化生态学的模型系统。为了研究这些相互作用，我们建立了一种体外培养单一寄生虫株的方法。此外，还开发了一种通过光密度 (OD) 测量来确定培养物中细胞数量的高通量方法。体外培养的方案允许在琼脂平板和培养基上培养不同的菌株。已经开发出细胞数与 OD 之间关系的校准曲线。随后，记录了不同基因型的 C. bombi 的生长速率。这些基因型之间的生长速率和世代时间存在显著差异。由于这可能与寄生虫的毒力有关，因此可以通过体内生长速率测定来证实这种关系。与传统的细胞计数相比，OD 测量的应用允许进行高通量实验，因为记录每个样本所需的时间减少了 30 倍。体外培养方法允许进行受控感染实验，以研究宿主-寄生虫相互作用。
