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目的了解广西横县鱼类吸虫感染的虫种,评价形态学结合ITS2序列分析鉴定囊蚴的有效性。方法采集横县某水库小鱼,采用人工消化法消化收集囊蚴,在解剖镜下分捡单个囊蚴,按形态学特征鉴定分类,提取不同形态囊蚴的DNA后进行ITS序列PCR扩增和测序,获得的序列在NCBI上进行Blast分析,并用Mega 6.0.6软件构建系统发生树。结果检测小型鱼类112条,吸虫囊蚴阳性率为39.29%。观察到5种不同形态的囊蚴。ITS2序列分析鉴定为4种吸虫:华支睾吸虫(Clonorchis sinensis)、钩棘单睾吸虫(Haplorchis pumilio)、台湾棘带吸虫(Centrocestus formosanus)和日本全冠吸虫(Holostephanus dubinini)。其中一种在形态学上判定为非人类吸虫的囊蚴经分子生物学鉴定为华支睾吸虫。结论在横县某水库小鱼中检出4种吸虫囊蚴,分别为华支睾吸虫、台湾棘带吸虫、钩棘单睾吸虫和日本全冠吸虫。仅靠形态学上对鱼类吸虫囊蚴进行虫种鉴定有一定的缺陷,可以结合ITS2序列分析来完善。
Objective To understand the species of fish flukes infected by fish in Hengxian County, Guangxi Province, and to evaluate the validity of the metabonomic analysis in combination with ITS2 sequence analysis. Methods The small fish in a reservoir of Hengxian County were collected and digested by manual digestion to collect metacercariae. Single metacercariae were collected under the dissection microscope. DNAs of different morphological characteristics were identified and classified according to morphological characteristics. The sequences obtained were subjected to Blast analysis on NCBI and the phylogenetic tree was constructed using Mega 6.0.6 software. Results A total of 112 small-sized fish were detected, and the positive rate of metacercariae was 39.29%. Five different forms of metacercaria were observed. ITS2 sequence analysis identified four species of trematodes: Clonorchis sinensis, Haplorchis pumilio, Centrocestus formosanus, and Holostephanus dubinini. One of the morphologically determined metacercariae of non-human trematodes was identified by molecular biology as Clonorchis sinensis. Conclusion Four species of metacercariae were found in the fish of a certain reservoir in Hengxian County. They were Clonorchis sinensis, Acanthoeba schrenckii, Monoclonal hookworm and Clonorchis japonicus. Only morphological identification of fish species of metacercariae larvae have some defects, can be combined with ITS2 sequence analysis to improve.