Prevalence and gene polymorphism analysis of Echinococcus granulosus in cattle and sheep in part areas of Xinjiang

doi:10.12140/j.issn.1000-7423.2022.05.006

Abstract

Abstract:

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

CLC Number:

R532.32

GUO Lu, WU Xiao-xia, DUAN Lan-li, WANG Bing-jie, XU Ning, AREAI Ahatai, WU Yun-hua, ZHAO Li, BAN Wan-li, CHEN Yun-ying, YU Wan-rong, LIU Shuai, PAN Xing-yu, WULIJIANG Kamali, XU Jing, MUNILA Teliewuhan, ZHANG Zhuang-zhi. Prevalence and gene polymorphism analysis of Echinococcus granulosus in cattle and sheep in part areas of Xinjiang[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2022, 40(5): 603-609.

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县（市） County (City)	羊 Sheep			牛 Cattle
县（市） County (City)	肝 Liver	肺 Lung	肝、肺 Liver, lung	肝 Liver	肺 Lung	肝、肺 Liver, lung
昭苏 Zhaosu	10	2	2	-	-	-
伊宁 Yining	13	7	16	3	4	6
博乐 Bole	12	11	8	2	3	0
特克斯 Turks	1	0	0	0	1	0
阿勒泰 Altay	2	0	0	-	-	-
塔城 Tacheng	2	7	0	-	-	-
额敏 Emin	3	1	4	0	1	0
合计 Total	43	28	30	5	9	6

突变种类 Mutation species	变异位点 Variable site
T-C 转换 T-C transition	15,120,238,269,273,296,300,313,336
C-T 转换 C-T transition	68,78,95,104,123,186
G-A 转换 G-A transition	258,322,324
A-G 转换 A-G transition	271
T-A 颠换 T-A transversion	315
T-G 颠换 T-G transversion	315,336