捻转血矛线虫离体培养的研究

本实验研究了:(1)三种人工合成培养基:M199、NCTC109、F_(12);(2)小牛脱纤维蛋白血(以下简称小牛脱纤血)、Filde’s试剂,Bovine hemin;(3)pH值等几种因素对离体培养中捻转血矛线虫(Haemonchou contortus)L_3发育至L_4的影响。结果表明:(1)在相同条件下,NCTC109和M199培养系中的L_3可发育至L_4晚期,F_(12)中L_3只发育至L_4早期;三种培养系中L_3发育至L_4的虫体数占存活虫体数的90％以上,以NCTC109中的发育率为最高,达100％。(2)小牛脱纤血、Filde’s试剂和Bovine hemin对L_3发育至L_4均有促进作用,虫体的存活率明显提高,晚四期幼虫的存活时间延长;小牛脱纤血和Filde’s试剂可使L_3至晚L_4的时间缩短。(3)pH 5.8～6.2时,L_3发育至L_4;速度明显减慢,发育率不到50％;pH 6.6～6.8时,L_3发育至L_4的同步性及发育率明显提高,以pH6.6为最佳。

IN VITRO CULTIVATION OF PARASITIC STAGE LARVAE OF HAEMONCHUS CONTORTUS (RUD, 1803)

The present paper represents the results of in vitro cultivation of parasitic stage larvae of H. contortus. The following are the main points: The influence of three commercial media-NCTC109, M199, F12; calf defibrinated blood, Fildes reagent, Bovine hemin; and the pH of the media on the development of H. contortus from exsheathed infective larvae to the fourth stage in vitro has been researched. The results indicate. (1) Under equal status, more than 90%L3 developed to L4 in NCTC109, M199 or F12 supplemented with CEE50 and CBS, but the larvae in medium F12 could not developed a step further, and stopped at the early fourth stage. In CNTC109 and M199, the larvae developed to late fourth stage, and NCTC109 was better than M199. (2) The medium NCTC109 + CEE50 + CBS supplemented with calf defibrinated blood, Fildes reagent or Bovine hemin was found to promote the development of L3 to L4 and increase the survival rate of L4. The calf defibrinated blood and Fildes reagent had the function to encourage the development of the larvae to the late fourth stage. On the contrary, Bovine hemin seemed to have no significant effect on the further development of L4. (3) In various cultural systems, when the pH value was 5.8-6.2, no more than 50% L3 could develop to L4 At pH 6.6-6.8, the synchronization of development and the yield of L4 were promoted. The optimum pH value was 6.6.