泡球蚴长期保存后生物活性的研究

目的继发性泡球蚴经液氮冻存后传代健康小鼠,观察其体内泡球蚴的生长情况并判定其生物活性,以探讨泡球蚴保种传代的新方法。方法将继发性感染泡球蚴的小鼠,液氮冻存7d后,常规复苏剖取包囊接种健康小鼠,生长两个月后颈椎脱臼处死,剖检并与常规接种方法相比较,从以下方面评价冻存效果:(1)大体观察及测量包囊湿重;(2)普通光学显微镜及透射电子显微镜观察组织学结构及超微结构;(3)检测包囊囊液中总蛋白含量、血清中特异性免疫球蛋白活动状态等免疫学指标;(4)检测血清碱性磷酸酶及囊液中钙离子含量等生化指标。结果液氮低温冻存后泡球蚴所接种试验组小鼠与常规接种组小鼠体内泡球蚴在包囊湿重、包囊组织结构、囊液蛋白及钙离子含量、血清碱性磷酸酶及特异性免疫球蛋白活动状态等方面均无显著差异(P>0.05)。结论液氮冻存后泡球蚴仍具有感染小鼠的能力,且接种后所生长泡球蚴组织与常规接种泡球蚴组织生物活性无显著差异。液氮冻存可以长期保存泡球蚴组织的活性。

The bioactivity evaluation of Echinococcus multilocularis after prolonged preservation

The bioactivity of the preserved Echinococcus multilocularis in liquid nitrogen (cryopreservation) was evaluated by infecting the cryopreserved materials into healthy mice and observing the growth of E. multilocularis and their bioactivities in order to investigate the novel method for cryopreservation of this worm in vitro. In this study, mice infected with E.multilocularis were preserved under liquid nitrogen condition for 7 days, and the favorite cysts were picked out from the frozen mice to inoculate into healthy mice. Two months later mice were sacrificed by cervical dislocation , dissected and several evaluations were carried out to compare the differences with the routine inoculation method. The effects of cryopreservation were evaluated in various aspects, such as (1): gross observation and measurement of wet weight of cysts, (2): histological structure and ultra-microstructure observation under light microscopy and transmission electro-microscopy; (3): total protein amount in hydatid cyst fluid (HCF) and the activities of specific immunoglobulins in serum, (4) contents of serum alkaline phosphatase (AKP) and Ca2+ ion in HCF. It was demonstrated that there was no significant difference in wet weight of cysts, histological structure and ultra-microstructure of cysts, the serum total protein, AKP and immunoglobulin content as well as the content of Ca2+ ion in HCF between these two groups of mice. These results suggest that the cryopreservation can maintain the activities of E.multilocularis over a long period of time.