旋毛虫不同虫期收集方法的比较

Comparison of collection methods of Trichinella spiralis bodies in various developmental stages

为了满足旋毛虫研究中对收集到的不同虫期虫体的数量、活力、纯净度等质量需求,基于相关文献中不同虫期虫体的收集方法,进行适当优化,比较了各收集方法的耗时长短、难易程度、收集的虫体数量及其感染力等,并总结出应用于不同实验需求的最适收集方法。同时,通过荧光定量PCR方法检测旋毛虫新生幼虫期特异性基因T668(AF331160.1)在不同时期各组织器官中的表达量,来间接反映移行期新生幼虫的含量,为后续描绘新生幼虫在动物体内的移行路径以及移行幼虫的研究提供新思路。研究结果表明:用贝尔曼静置法收集的肌幼虫的活力和侵袭力最强,用贝尔曼搅拌法收集的肌幼虫死虫数量最少,以生理盐水作为消化液更佳,用常规孵育装置收集法收集的成虫的数量最多,用离心法收集的新生幼虫的数量最多,用自然沉淀法收集的新生幼虫的感染力最强;旋毛虫雌虫在回肠和攻虫后第5天时数量最多;第4天时肺和后腿肌中移行期新生幼虫含量较多,第8天时在心、肾、舌肌、膈肌中较多,第12和15天时在心中最多。研究结果为满足不同实验需求,提供了虫体的最适收集方法。

In order to address the quality requirements for the quantity,viability and purity in the study of Trichinella spiralis at different stages,this research aimed to compare the length of time-consuming,the extent of difficulty,the amount of collection nematode and force of infection of different collection methods obtained from the relevant literature,and then optimize and summarize the most suitable collection method for varying experiments.At the same time,the expression of the specific gene T668(AF331160.1) of T.spiralis newborn larvae in different tissues and organs at different stages detected by RT-qPCR was purposed to indirectly reflect the content of the migrating newborn larvae,and provide a new idea for the subsequent description of the migratory path of the newborn larvae in host and the research of the migratory larvae.The results showed that the muscle larvae collected by Baermann’s standing method presented a highest viability and invasiveness,and Baermann’s mixing round method led to the least larval deaths,using physiological saline as digestive solution to collect worms was more favorable.The most numerous adult worms and newborn larvae could be obtained by conventional incubation device and centrifugal method,respectively.The newborn larvae collected by natural precipitation method showed the strongest force of infection.Furthermore,female T.spiralis could be observed the maximum number in ileum on day 5 post infection.Migrating newborn larvae could be detected a higher number in the lung and hindleg muscles at 4 d,in the heart,kidney,tongue muscle and diaphragm at 8 d,and in the heart at 12 d and 15 d,respectively.The findings provided the most appropriate collection method to satisfy different experimental requirements.