Abstract：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.
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