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目的分析安徽省湖沼型血吸虫病流行区林业血防工程区钉螺细胞色素氧化酶Ⅰ(COⅠ)基因的差异性,探讨林业血防工程中不同生态环境对钉螺生态的影响。方法采集长江安徽段上游安庆、中游铜陵、下游无为等3地林业血防工程区和草滩对照点钉螺,提取DNA,PCR扩增COⅠ基因并测序。采用blast对COⅠ基因比较相似性。采用微小钉螺、双脐螺COⅠ基因序列作为外群参照序列,Kimura双参数法计算遗传距离,MEGA 3.1软件非加权组平均法(UPGMA)和邻近法(NJ)构建系统发生树。结果 PCR扩增获得的COⅠ基因约为700 bp(含两端引物)。各地林业工程区之间,各林业工程区与其草滩对照点之间的COⅠ基因相似性均较高,均≥98%;安庆、铜陵、无为等3地林业工程区之间遗传距离为0.003～0.012,3地林业工程区与草滩对照点的遗传距离分别为0.003、0.019、0.007。2种方法构建的进化树拓扑结构基本一致。进化树分2大分支,双脐螺独立成一大支,其他的成一大支(在这大支中微小钉螺又成一支),3地的林业工程区与对照草滩钉螺基因差异比较小。结论长江安徽段上、中、下游林业血防工程区钉螺COⅠ基因在不同的生态环境下可能已存在一定程度的遗传分化,但分化程度较低。
Objective To analyze the differences of cytochrome oxidase Ⅰ (COⅠ) gene in snail schistosomiasis prevention project of schistosoma japonicum endemic areas in Anhui province and to explore the ecological effects of different ecological environments on Snail ecologies in forestry blood protection projects. Methods The snail was collected from the control sites of Anqing, Zhongling, Tongling, and Wuwei in the middle reaches of the Yangtze River. The COⅠ gene was amplified by PCR and sequenced. Blast was used to compare the COI genes. The phylogenetic tree was constructed by using COⅠ gene sequence of Micromonas spiraea and Umbilical cord as reference sequence of outer group, Kimura’s two-parameter method to calculate genetic distance, MEGA 3.1 software unweighted group average method (UPGMA) and neighboring method (NJ). Results The COⅠ gene obtained by PCR amplification was about 700 bp (containing two - terminal primers). The COⅠgene similarity among the forestry engineering areas and between the forestry engineering areas and their grassland control points were all higher than 98%. The genetic distance between the three forestry engineering areas in Anqing, Tongling and Wuwei was between 0.003 and 0.012 , And the genetic distance of the three control sites of forestry project and grassland were respectively 0.003, 0.019 and 0.007.2. The topological structure of the evolutionary tree was basically the same. The phylogenetic tree is divided into two major branches, the two umbilical snails independently become a major branch, the others become a major branch (in this branch, the minor snails become one), and the difference between the snail genes in the three forestry areas and the control grassland is relatively small. Conclusion The COⅠ gene of Oncomelania snails in the upper, middle and lower reaches of the Yangtze River in Anhui Province may have a certain degree of genetic differentiation in different ecological environments, but the degree of differentiation is low.