骆驼源细粒棘球绦虫成虫体外培养的观察

Observations on Culture in vitro of Protoscolices of Echinococcus granulosus from Camel

本文报告建立细粒棘球绦虫成虫体外培养的方法和结果.以凝固小牛血清做为固相,以含有酵母浸液、葡萄糖和牛磺胆酸钠等补充成分的199培养基为液相的双相培养基可以支持寄生虫的生长并向成虫方向发育。经胃蛋白酶消化和犬胆汁处理后外翻的原头节于培养第6天出现排泄管,第15天形成排泄囊,第16天出现分带,第36和39天明显形成两节的虫体。第60天可见到长成第二体节的成虫,使整个虫体形成3节,但此后虫体开始迅速崩解,至第75天后完全崩解。于培养不同时间测量虫体长度,在培养45天时发育良好的虫体长度范围为717～919.1μm,平均815.22±63.54μm。在第29天和第41天出现生殖原基和生殖孔,但生殖器官未见明显发育。作者认为细粒棘球绦虫成虫体外培养可发育形成三节成虫,并维持75天,表明这种体外培养系统能够支持原头节向成虫方向发育.生殖器官发育延缓可能提示发育后期所需营养物质供应不足。细粒棘球绦虫成虫体外培养的初步成功,为国内对该种蠕虫的有关学科领域的研究工作提供了实验工具。

This paper reports the method and results of establishing culture in vitro of Echinococcus granulosus.Double-phase medium consisting of coagulated calf serum as solid phase and medium 199 containing yeast extract,glucose and sodium taurocholate as liquid phase al- lowed the growth of the parasite and the deve lopment towards adults.In protoscolices with extroversion following digestion with pepsin and treatment with dog's bile,excretory duct appeared on day 6 after incubation,excretory bladder on day 15,zonation on day 16,and worms with two definite segments on days 36 and 39.Adult worms with the second body segment were visible on day 60,thus forming the whole worm with 3 segments.Disintegra- tion of the worm,however,occurred rapidly afterwards;and complete disintegration oc- curred on day 75.Measurements of worm length at different time points of incubation showed that on day 45 of incubation welldeveloped worms measured 717-919.1 micron long,with a mean of 815.22±63.54 micron.Genital rudiment and gonopore occurred on days 29 and 41,respectively,but genitals were not well developed.The formation of adult worms with three segments following culture in vitro of protoscolices of E.granulosus and the maintenance for as long as 75 days suggest that this culture system in vitro allows the devel- opment of protoscolices towards adult worms,and the retarded development of genitals may indicate the insufficient supply of nutrients required in the later stage of development.The preliminary success in culture in vitro of adult worms of E.granulosus might be instrumental to the research work in the field of related subjects on this tapeworm in this country.