Lanuza M D, Carbajal J A, Villar J, Borrás R
Departamento de Microbiología, Facultad de Medicina y Hospital Clínico Universitario de Valencia, Spain.
Parasitol Res. 1997;83(1):60-3. doi: 10.1007/s004360050209.
An improved method for Blastocystis hominis culture and axenization was developed in the present study. Stool samples were cultured in prereduced Boeck-Drbohlav NHI modified medium (with several modifications) supplemented with antibiotics (0.4% ampicillin, 0.1% streptomycin, 0.0006% amphotericin B). Axenization was performed by the combination of partial purification of B. hominis by Ficoll-metrizoic acid gradient and inoculation in fresh medium containing active antibiotics against remaining bacteria. A total of 25 strains were obtained by this procedure. The time required for axenization ranged between 3 and 5 weeks. The generation time of axenic strains ranged from 6.6 to 12.1 h (mean +/- SD 110.0 +/- 1.8 h) and the mean number of generations was 2.5 +/- 0.6 h per 24 h. The size of vacuolar and ameboid forms found in stools and in culture was similar. The special formulation of the medium used reduced the generation time and did not modify the cellular size as compared with fecal forms.
本研究开发了一种改进的人芽囊原虫培养和无菌培养方法。粪便样本在预先还原的改良Boeck-Drbohlav NHI培养基（有若干修改）中培养，并添加抗生素（0.4%氨苄青霉素、0.1%链霉素、0.0006%两性霉素B）。通过Ficoll-泛影酸梯度对人芽囊原虫进行部分纯化，并接种到含有针对残留细菌的活性抗生素的新鲜培养基中，从而进行无菌培养。通过该程序共获得25株菌株。无菌培养所需时间为3至5周。无菌菌株的代时为6.6至12.1小时（平均±标准差为110.0±1.8小时），每24小时的平均代数为2.5±0.6小时。在粪便和培养物中发现的空泡型和阿米巴样形态的大小相似。与粪便形态相比，所用培养基的特殊配方缩短了代时，且未改变细胞大小。
