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目的观察血吸虫活虫卵在不同保存液中的保存时间及孵化毛蚴的活力,获得血吸虫活虫卵最佳保存条件,探索用作孵化参考品的可行性。方法收集人工感染血吸虫家兔直肠粪便,经80、100、160孔/25.4 mm和200孔/25.4 mm分样筛过滤去除粗粪渣,260孔/25.4 mm收集血吸虫活虫卵,于0.9%Na Cl溶液、1.2%Na Cl溶液、磷酸盐缓冲液(PBS,p H7.2)、1.0%蔗糖溶液和Mili-Q水5种保存液中混匀,分别于4℃和15℃恒温保存。每隔7 d对保存虫卵进行孵化,观察毛蚴活力并计数。结果在4℃条件下,1.0%蔗糖溶液和1.2%Na Cl溶液中虫卵孵化率分别于49 d后和126 d后降至0;0.9%Na Cl溶液和PBS保存液中虫卵孵化率于112 d后降至10%以下,140 d后孵出的毛蚴活力较差;Mili-Q水保存的虫卵孵化率于196 d后降至10%以下,280 d后孵出毛蚴,活力明显下降且不利于观察。在15℃条件下,各保存液中虫卵的孵化率分别在49~105 d之间逐渐降至0。结论实验室以Mili-Q水为血吸虫活虫卵保存液,置4℃恒温保存,196 d之内的冷藏虫卵具有用作孵化阳性参考品的价值。
Objective To observe the preservation time of Schistosoma japonicum eggs in different preservation solutions and the viability of hatching miracidia to obtain the optimal storage conditions for live schistosome eggs and to explore the feasibility of using them as hatching reference materials. Methods Colorectal mucosa of rabbits with artificial infection of schistosomiasis was collected, crude cohort was removed by filtration through 80, 100, 160, 25.4 and 200-hole / 25.4-mm sieve sieves. Cl solution, 1.2% NaCl solution, phosphate buffer solution (PBS, p H7.2), 1.0% sucrose solution and Mili-Q water and stored at 4 ℃ and 15 ℃ respectively. Incubate the preserved eggs every 7 days and observe the activity and count of the miracidia. Results At 4 ℃, the hatching rates of eggs in 1.0% sucrose solution and 1.2% NaCl solution decreased to 0 after 49 days and 126 days respectively. The hatching rate of eggs in 0.9% NaCl solution and PBS After 112 days, it decreased to less than 10%. After 140 days, the maggots hatched were less viable. The egg hatching rate of Mili-Q water was reduced to below 10% after 196 days. The maggots were hatched after 280 days and their viability significantly decreased And not conducive to observation. Under the condition of 15 ℃, the hatching rate of eggs in each preservation solution decreased to 0 between 49 ~ 105 d respectively. Conclusion The Mili-Q water was used as the preservation solution of Schistosoma japonicum live egg and stored at 4 ℃. The frozen eggs within 196 days had the value of hatching positive reference.