收稿日期: 2022-02-22
修回日期: 2022-05-11
网络出版日期: 2022-10-28
基金资助
国家自然科学基金(31560692)
Prevalence and gene polymorphism analysis of Echinococcus granulosus in cattle and sheep in part areas of Xinjiang
Received date: 2022-02-22
Revised date: 2022-05-11
Online published: 2022-10-28
Supported by
National Natural Sciecne Foundation of China(31560692)
目的 了解新疆维吾尔自治区（简称新疆）部分地区牛、羊细粒棘球蚴感染情况和基因多态性,为该地区细粒棘球蚴病的防治提供数据参考。 方法 于2020—2021年从伊宁市、阿勒泰市、塔城市、昭苏县、特克斯县、额敏县、博乐市的定点屠宰场采集屠宰后牛、羊的肝脏和肺脏,通过肉眼观察与触摸初步判定感染情况,采集疑似细粒棘球蚴包囊并提取DNA,PCR扩增细粒棘球绦虫细胞色素氧化酶1（cox1）基因并测序。从GenBank中下载细粒棘球绦虫cox1基因G1、G3、G4、G5、G6型序列,与测定的序列行BLAST比对。利用MEGA 7.0软件用邻接法构建系统进化树,用Popart软件构建cox1基因单倍型网络图,用DnaSPv5软件分析cox1基因多态性位点。使用SPSS17.0软件进行统计学分析,对不同地区牛、羊及不同脏器感染率的比较采用χ2检验。 结果 新疆7个县（市）共调查4 977头（只）牛、羊（牛563头、羊4 414只）,采集到疑似细粒棘球蚴包囊141份,其中121份扩增出850 bp的条带并测序成功。牛、羊总感染率为2.07%（103/4 977）,其中博乐市牛、羊的感染率最高（6.50%,32/492）,与伊宁市（1.80%,38/2 108）、塔城市（1.14%,9/790）、特克斯县（1.06%,2/189）、额敏县（0.72%,7/970）相比,感染率差异有统计学意义（χ2 = 49.873、33.682、8.762、28.666,均P < 0.05）。牛、羊的感染率分别为3.02%（17/563）、1.95%（86/4 414）,两者差异有统计学意义（χ2 = 14.296,P < 0.05）。共采集羊肝、肺包囊101份,其中肝包囊占42.57%（43/101）,肺包囊占27.72%（28/101）,肝、肺合并感染包囊占29.70%（30/101）。共采集牛肝、肺包囊20份,其中肝包囊占25.00%（5/20）,肺包囊占45.00%（9/20）,肝、肺合并感染包囊占30.00%（6/20）。BLAST比对结果显示,121条序列中120条（99.2%）与细粒棘球绦虫G1基因型序列（MN886258.1、MG280957.1、KX020359.1和MG672143.1）的同源性为98%~100%,在系统进化树上聚为一支;1条（0.8%）与G3基因型序列（MG682532.1）的同源性为100%,在系统进化树上聚为一支。单倍型分析结果显示,cox1基因序列共有18个单倍型,Hap 1~Hap 17为G1型,Hap 18为G3型,其中Hap 1为最大的单倍型群体;18个单倍型间共有20个变异位点。序列比对结果显示,121条序列中有52条出现了碱基的转换和颠换。 结论 新疆7个县（市）牛、羊细粒棘球蚴感染率存在明显差异,感染细粒棘球蚴的基因型为G1（99.2%）、G3（0.8%）,有18个单倍型,Hap 1为主要致病单倍型。
郭璐 , 吴小霞 , 段兰利 , 王冰洁 , 徐宁 , 阿热艾·阿哈太 , 乌云花 , 赵莉 , 班万里 , 陈云英 , 余婉蓉 , 刘帅 , 潘星羽 , 吾力江·卡马力 , 徐晶 , 穆尼拉·特列吾汗 , 张壮志 . 新疆部分地区牛、羊细粒棘球蚴感染情况调查及基因多态性分析[J]. 中国寄生虫学与寄生虫病杂志, 2022 , 40(5) : 603 -609 . DOI: 10.12140/j.issn.1000-7423.2022.05.006
Objective To understand the prevalence and gene polymorphism of Echinococcus granulosus in cattle and sheep in part areas of Xinjiang Uygur Autonomous Region (Xinjiang), and to provide field data for the prevention and control of cystic echinococcosis in this region. Methods From 2020 to 2021, the livers and lungs of the slautered cattle and sheep were collected from the designated slaughterhouses in Yining City, Altay City, Tacheng City, Zhaosu County, Turks County, Emin County and Bole City. The infection was preliminarily assessed by visual inspection and palpation. Suspected E. granulosus cysts were collected for DNA extraction, of which the E. granulosus cytochrome oxidase 1 (cox1) gene was amplified by PCR and sequenced. The sequences obtained were BLAST aligned with the sequences of G1, G3, C4, G5 and G6 downloaded. The phylogenetic tree was constructed by the neighbour-joining method using MEGA 7.0 software, the haplotype network of cox1 gene was constructed by Popart software, and the gene polymorphism sites were analyzed by DnaSPvS software. SPSS17.0 software was used for statistical analysis. The infection rates of cattle, sheep and different organs in different regions were compared by Chi-square (χ2) test. Results A total of 4 977 livestock (563 cattle, 4 414 sheep) were investigated in 7 counties (cities) in Xinjiang, from them 141 cysts were collected. An amplicon of 850 bp was found in 121 cysts, and was successfully sequenced. The overall infection rate in cattle and sheep was 2.07% (103/4 977), among them the highest infection rate in cattle and sheep was seen in Bole City (6.50%, 32/492) compared to Yining City (1.80%, 38/2 108), Tacheng City (1.14%, 9/790), Turks County (1.06%, 2/189) and Emin County (0.72%, 7/970). The difference was statistically significant (χ2 = 49.873, 33.682, 8.762, 28.666, all P < 0.05). The infection rates of cattle and sheep were 3.02% (17/563) and 1.95% (86/4 414), respectively, and the difference was statistically significant (χ2 = 14.296, P < 0.05). The liver and lung cysts from 101 sheep were collected, in which liver cysts accounted for 42.57% (43/101), lung cysts accounted for 27.72% (28/101), and liver and lung co-infection cysts accounted for 29.70% (30/101). The liver and lung cysts from 20 cattle were collected, in which liver cysts accounted for 25.00% (5/20), lung cysts accounted for 45.00% (9/20), and liver and lung co-infection cysts accounted for 30.00% (6/20). BLAST alignment showed that 120 (99.2%) of the 121 sequences had 98%-100% homology with the G1 genotype sequence of E. granulosus (MN886258.1, MG280957.1, KX020359.1 and MG672143.1), which clustered into one branch on the phylogenetic tree, and One (0.8%) had 100% homology with the G3 genotype sequence (MG682532.1), which clustered into one branch on the phylogenetic tree. Haplotype analysis showed that there were 18 haplotypes in cox1 gene sequence, Hap 1-Hap 17 was G1 genotype, Hap 18 was G3 genotype. Hap 1 had the largest haplotype population. There were a total of 20 variation sites among 18 haplotypes. Sequence alignment showed that 52 of 121 sequences had base conversion and transversion. Conclusion The E. granulosus infection rates among cattle and sheep in 7 counties (cities) in Xinjiang are significantly different. G1 (99.2%) and G3 (0.8%) genotypes and 18 haplotypes are found, among which Hap 1 is the dominant pathogenic haplotype.
Key words： Cystic echinococcosis; cox1 gene; Haplotype
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