云南腾冲中缅边境牛羊硬蜱形态学鉴定及基因多态性分析

doi:10.16431/j.cnki.1671-7236.2025.10.033

摘要/Abstract

摘要： 【目的】调查云南腾冲中缅边境牛羊硬蜱种类和基因多样性，为其所致疾病的防控提供依据。【方法】于2024年6－7月调查腾冲市4个乡镇牛羊硬蜱感染情况，采集硬蜱并对其进行形态学鉴定；选择16只硬蜱(牛源和羊源各8只)为研究对象，通过PCR扩增其线粒体cox1基因和核糖体ITS2基因序列，通过DNAStar、Mega 7.0和DnaSP软件分析其遗传多样性、单倍型多态性和种系发育关系。【结果】调查的4个乡镇中牛羊感染硬蜱的场阳性率均为100% (24/24)；其中牛和羊硬蜱感染率分别94.37%(64/71)和91.74%(111/121)；共采集硬蜱362只，形态学鉴定显示均为微小扇头蜱；16只牛羊源微小扇头蜱cox1(762 bp)和ITS2基因序列(1 477 bp)种内差异分别为0～0.5%和0～1.3%；与GenBank收录的微小扇头蜱(分支C)cox1和ITS2基因序列相似性分别超过99.5%和99.0%；cox1和ITS2基因的单倍型多样性分别为0.858和0.932，而核苷酸多样性分别为0.00395和0.00514。遗传进化树分析发现，16只微小扇头蜱与已知微小扇头蜱均属于同一分支，不同宿主源分离株在该分支中随机分布，但与其他硬蜱所属分支相距较远。【结论】云南腾冲牛羊体表携带硬蜱情况严重，且均为微小扇头蜱，属于分支C；不同宿主源微小扇头蜱具有一定的遗传变异，但单倍型多样性和核苷酸多样性均较低，且未出现遗传分化现象。

关键词: 牛; 羊; 微小扇头蜱; cox1基因; ITS2基因; 遗传进化; 单倍型多样性

Abstract: 【Objective】 The aim of this study was to investigate the species and genetic diversity of hard ticks from cattle and sheep in the China-Myanmar border region of Tengchong city,Yunnan province,and provide a basis for the prevention and control of diseases caused by them.【Method】 The present study conducted a survey on tick infection in cattle and sheep in four towns in Tengchong city from June to July 2024,the ticks were collected and submitted to morphological identification.The mitochondrial cox1 gene and ribosomal ITS2 gene sequences of 16 ticks (8 from cattle and 8 from sheep) were amplified by PCR,their genetic diversity,haplotype polymorphism and phylogenetic relationships were analyzed using DNAStar,Mega 7.0 and DnaSP software.【Result】 The farm positive rate of tick infection in cattle and sheep in the four investigated towns was 100% (24/24).The infection rates of ticks in cattle and sheep were 94.37% (64/71) and 91.74% (111/121),respectively.A total of 362 ticks were collected,all of which were identified as Rhipicephalus microplus through morphological analysis.The intraspecific differences in cox1 (762 bp) and ITS2 (1 477 bp) gene sequences of 16 R.microplus isolates were 0-0.5% and 0-1.3%,respectively.The similarity between cox1 and ITS2 gene sequences of R.microplus (branch C) in GenBank exceeded 99.5% and 99.0%,respectively.The haplotype diversity of cox1 and ITS2 genes was 0.858 and 0.932,respectively,while the nucleotide diversity was 0.00395 and 0.00514,respectively.The results of phylogenetic tree analysis revealed that all 16 R.microplus belonged to the same branch as known R.microplus,the isolates from different host sources were randomly distributed in this branch,but they were far away from the branches of other ticks.【Conclusion】 The prevalence of ticks on the surface of cattle and sheep in Tengchong city,Yunnan province was severe,and all ticks were R.microplus belonging to branch C.The R.microplus strains from different hosts had certain genetic variations,but the haplotype diversity and nucleotide diversity were low,and there was no genetic differentiation phenomenon.

Key words: cattle; sheep; Rhipicephalus microplus; cox1 gene; ITS2 gene; genetic evolution; haplotype diversity

中图分类号:

S852.74⁺6

段雨晴, 王金萍, 余洋, 孙若迪, 杜月仙, 赵国稳, 胡本亮, 彭明正, 谭磊, 姚俊. 云南腾冲中缅边境牛羊硬蜱形态学鉴定及基因多态性分析[J]. 中国畜牧兽医, 2025, 52(10): 4884-4893.

DUAN Yuqing, WANG Jinping, YU Yang, SUN Ruodi, DU Yuexian, ZHAO Guowen, HU Benliang, PENG Mingzheng, TAN Lei, YAO Jun. Morphological Identification and Genetic Polymorphism Analysis of Hard Ticks from Cattle and Sheep in the China-Myanmar Border Region of Tengchong City,Yunnan Province[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(10): 4884-4893.

参考文献

[1] 徐兴莉，杨虎.江西部分地区微小扇头(牛)蜱中肠优势菌群分析[J].中国动物传染病学报，2023,31(1):159-165. XU X L,YANG H.Investigation of dominant microbiota in the midguts of Boophilus microplus in some regions of Jiangxi province[J].Chinese Journal of Animal Infectious Diseases,2023,31(1):159-165.(in Chinese)
[2] 潘海钰，张雪玲，麻林坤，等.中国七省(区)牛蜱的种类鉴定及其携带无形体的分子检测[J].湖南农业大学学报(自然科学版)，2023,49(6):723-729. PAN H Y,ZHANG X L,MA L K,et al.Identification of tick species in cattle and detection of Anaplasma spp.in these ticks in 7 provinces and regions of China[J].Journal of Hunan Agricultural University (Natural Sciences),2023,49(6):723-729.(in Chinese)
[3] 于莲琪，张本光，盛兆安.山东主要蜱种分布现状及蜱携病原的研究进展[J].中国卫生杀虫药械，2024,30(2):185-190. YU L Q,ZHANG B G,SHENG Z A.Distribution status of major tick species and research progress on tick-borne pathogens in Shandong[J].Chinese Journal of Hygienic Insecticides & Equipments,2024,30(2):185-190.(in Chinese)
[4] 曾予，刘志云，段立科，等.江西省吉安市微小扇头蜱携带病原菌检测及其分型研究[J].中国媒介生物学及控制杂志，2024,35(3):381-388. ZENG Y,LIU Z Y,DUAN L K,et al.Detection and genotyping of pathogenic bacteria carried by Rhipicephalus microplus in Ji’an,Jiangxi province,China[J].Chinese Journal of Vector Biology and Control, 2024,35(3):381-388.(in Chinese)
[5] 吴胜春，孟娇，黄健胜，等.贵州省施秉县媒介蜱携带双压巴贝斯虫的调查研究[J].中国媒介生物学及控制杂志，2023,34(2):254-261. WU S C,MENG J,HUANG J S,et al.Investigation of Babesia bigemina carried by ticks in Shibing county,Guizhou province,China[J].Chinese Journal of Vector Biology and Control,2023,34(2):254-261.(in Chinese)
[6] OGOLA E O,KOPP A,BASTOS A D S,et al.Jingmen tick virus in ticks from Kenya[J].Viruses,2022,14(5):1041.
[7] 张琳，贺真，陆振华，等. 陕西部分地区长角血蜱的鉴定及其基因多态性[J].动物医学进展，2024，45(5)：1-7. ZHANG L, HE Z, LU Z H, et al. Identification and gene polymorphism of Haemaphysalis longicornis in some areas of Shaanxi province[J]. Progress in Veterinary Medicine, 2024,45(5): 1-7. (in Chinese)
[8] 李中波，侯强红，舒鸣，等. 不同地区褐黄血蜱的单倍型多态性、遗传分化和种系发育关系分析[J].中国兽医杂志，2023, 59(9): 60-66. LI Z B, HOU Q H, SHU M, et al. Haplotype diversity, genetic difference, and phylogenetic analysis of Haemaphysalis flava from different regions[J].Chinese Journal of Veterinary Medicine, 2023, 59(9): 60-66. (in Chinese)
[9] 沈雪鹰，蒋宝贵，何志海，等.云南省腾冲市牛、羊中无形体感染的分子流行病学调查[J].中国预防兽医学报，2020,42(10):1068-1072. SHEN X Y,JIANG B G,HE Z H,et al.Molecular epidemiological investigation on anaplasmosis in cattle and goat in Tengchong,Yunnan province[J].Chinese Journal of Preventive Veterinary Medicine,2020,42(10):1068-1072.(in Chinese)
[10] ZEB I,ALMUTAIRI M M,ALOUFFI A,et al.Low genetic polymorphism in the immunogenic sequences of Rhipicephalus microplus clade C[J].Vaccines (Basel),2022,10(11):1909.
[11] HORNOK S,FARKAS R,DUONG N N,et al.A morpho-phylogenetic update on ixodid ticks infesting cattle and buffalos in Vietnam,with three new species to the fauna and a checklist of all species indigenous to the country[J].Parasit Vectors,2024,17(1):319.
[12] 陈俊蓉，徐翠蓉，谢昕言，等.云南部分地区牛羊体表寄生虫的形态及分子鉴定[J].畜牧与兽医，2023,55(10):56-60. CHEN J R,XU C R,XIE X Y,et al.Morphological and molecular identification of parasitic ticks on cattle and sheep in some parts of Yunnan province[J].Animal Husbandry & Veterinary Medicine,2023,55(10):56-60.(in Chinese)
[13] 邓国藩，姜在阶.中国经济昆虫志：第39册[M].北京：科学出版社，1991. DENG G F,JIANG Z J.Economic Insect Fauna of China:Volume 39[M].Beijing:Science Publishing,1991.(in Chinese)
[14] 刘雨珂，王爱兵，刘国华，等.豪猪血蜱的形态学和分子生物学鉴定[J].中国兽医科学，2022,52(2):214-222. LIU Y K,WANG A B,LIU G H,et al.Morphological and molecular biological identification of Haemaphysalis hystricis[J].Chinese Veterinary Science,2022,52(2):214-222.(in Chinese)
[15] 李兰花，李胜国，王加志，等.腾冲县医疗机构门诊发热病人蜱虫叮咬情况调查[J].中国血吸虫病防治杂志，2016,28(3):323-326. LI L H,LI S G,WANG J Z,et al.Investigation of tick bites in outpatients with fever from health care facilities in Tengchong county[J].Chinese Journal of Schistosomiasis Control,2016,28(3):323-326.(in Chinese)
[16] WANG J,SHAH T,ZHOU J,et al.Identification,characterization,and homology analysis of a novel strain of the Crimean-Congo hemorrhagic fever virus from Yunnan,China[J].Microorganisms,2024,12(7):1466.
[17] JI H,CHANG L,YAN Y,et al.Pilot surveillance of Babesia,Borrelia burgdorferi,and Anaplasma phagocytophilum among Chinese blood donors-Xinjiang,Inner Mongolia,and Heilongjiang PLADs,China,2022-2023[J].China CDC Weekly,2024,6(46):1206-1210.
[18] QU L,LI X,HUANG B,et al.Identification and characterization of Jingmen tick virus in ticks from Yunnan imported cattle[J].Vector Borne and Zoonotic Diseases (Larchmont,N.Y.), 2023,23(5):298-302.
[19] WU Z,ZHANG M,ZHANG Y,et al.Jingmen tick virus:An emerging arbovirus with a global threat[J].mSphere,2023,8(5):e0028123.
[20] JIA N,LIU H B,NI X B,et al.Emergence of human infection with Jingmen tick virus in China:A retrospective study[J]. EBioMedicine,2019,43:317-324.
[21] 亚红祥，沈姝，苏正元，等.云南省西北地区家畜体表蜱类形态学与分子生物学鉴定[J].疾病监测，2017,32(7):609-614. YA H X,SHEN S,SU Z Y,et al.Morphological and molecular identification of tick species from body surface of domestic animals in Northwestern Yunnan[J].Disease Surveilllance,2017,32(7):609-614.(in Chinese)
[22] 谢昕言，陈俊蓉，徐翠蓉，等.云南部分地区家畜寄生蜱携带病原情况调查[J].动物医学进展，2023,44(8):131-135. XIE X Y,CHEN J R,XU C R,et al.Investigation on pathogens carried by ticks on livestock in partial areas of Yunnan[J].Progress in Veterinary Medicine, 2023,44(8):131-135.(in Chinese)
[23] 亚红祥，沈姝，苏正元，等.滇西横断山区家畜体表蜱类调查及鉴定[J].中国人兽共患病学报，2016,32(10):856-870. YA H X,SHEN S,SU Z Y,et al.Tick species and genetic variants analysis of tick gene in Hengduan mountains,West Yunnan province,China[J].Chinese Journal of Zoonoses,2016,32(10):856-870.(in Chinese)
[24] 亚红祥，张云智，王静林.云南临沧地区蜱种分子生物学鉴定及埃立克体基因序列分析[J].中国人兽共患病学报，2016,32(9):793-797. YA H X,ZHANG Y Z,WANG J L.Molecular identification of tick species and sequence analysis of Ehrlichia in Lincang area of Yunnan province,China[J].Chinese Journal of Zoonoses,2016,32(9):793-797.(in Chinese)
[25] LU M,MENG C,ZHANG B,et al.Prevalence of spotted fever group Rickettsia and Candidatus Lariskella in multiple tick species from Guizhou province,China[J]. Biomolecules,2022,12(11):1701.
[26] LI J,CHEN Z H,JIANG L,et al.Characterization of cattle-origin ticks from Southern China[J].Acta Trop,2018,187:92-98.
[27] ZHANG L,XU W,ZHAO Y,et al.Identification and characterization of Jingmen tick virus from Rhipicephalus microplus in Hunan,China[J]. Acta Tropica,2024,260:107378.