巨型艾美球虫第二代裂殖体的分离纯化

Isolation and purification of the second generation schizont of Eimeria maxima

目的寻找分离纯化巨型艾美球虫第二代裂殖体效率更高的方法。方法用孢子化的巨型艾美球虫卵囊喂食21～28日龄SPF雏鸡，平均50万个／只，72h后剖检出小肠中段，综合运用酶消化、离心洗涤、红细胞裂解和Percoll密度梯度离心等方法分离纯化巨型艾美球虫的第二代裂殖体。结果从35羽鸡的小肠中段中纯化到裂殖体4．1×10^5个，平均每羽鸡获得裂殖体1．17×10^5个，回收率约为89．6％。结论此方法简单易行，纯化回收结果适用于艾美属球虫分子生物学方面的研究。

Objective To find a better way for separating and purifying the second generation schizonts of Eimeria maxima. Methods Twenty-one-day-old to twenty-eight-day-old SPF chicken were inoculated with 50 000 ooeysts of Eimeria maxima per bird, respectively. The estimate of sehizont production was made on 72 h after inoculation. Enzymatic digestion, centrifugation-washing, red blood cell disruption and Percoll density gradient eentrifugation were synthetically applied to separate and purify the second generation schizonts of Eimeria maxima. Results We obtained 4.1 ×10^5 second generation sehizonts from the jejunum of 35 chicken after autopsy, on an average of 1.17 ×10^4 sehizonts each bird with a 89.6% recovery rate. of schizonts. Conclusion This method can be applied to the molecular biology studies on Eimeria maxima.