锥虫长期超低温保藏方法的研究

STUDIES ON THE LONG-TERM STORAGE METHOD AT ULTRA LOW-TEMPERATURE FOR TRYPANOSOMES

作者观测了不同保护剂、稀释液、pH值、降温方法、复苏温度、冻融次数、液相气相交替以及解冻后在普通冰瓶中存放时间对伊氏锥虫浙江虫株的感染性及致病力的影响。在此试验基础上,又对伊氏锥虫的6个不同虫株、媾疫锥虫、刚果锥虫、布氏锥虫等4个种的9个虫株,进行了超低温保藏试验和长期保藏效果观察。已测定的有效保藏期伊氏锥虫达574～3200天、媾疫锥虫达2866天、刚果锥虫达763天、布氏锥虫达783天。通过反复试验,提出了锥虫的常规保藏方法。

The effects of different cryoprotectant, diluent, pH value, hypothermia method, anabiosis temperature, freeze-thaw times, liquid-gas exchange and the time put in general ice bottle on the infectivity and pathogenicity of Zhejiang strain of Trypanosoma evansi were observed. On the basis of these experiments, long-term storage tests with 6 strains of T. evansi and T. equiperdum, T. congolense, T. brucei at ultra low-temperature were carried out, respectively. It was demonstrated that the effective storage periods were 574-3200 days for T. evansi, 2866 days for T. equiperdum, 763 days for T. congolense, 783 days for T. brucei, respectively. Through repeated experiments, a routine storage method has been established.