丁立,张力,刘全

• 1:佛山科学技术学院生命科学与工程学院
• 2:东北农业大学兽医学院黑龙江省动物疫病重点实验室

摘要(Abstract)：

蜱传病呈全球分布，危害人类和动物健康。为了研究湖南省娄底市现存蜱媒病原体，于2021—2022年在湖南省娄底市采集1 395只微小牛蜱样本，提取蜱样品基因组DNA。选取蜱线粒体COI基因扩增特异性的基因片段，对微小牛蜱进行鉴定。为了对该蜱携带的病原进行检测，采用PCR方法对立克次体ompA和gltA基因，无形体groEL和23S rRNA基因，埃立克体groEL和16S rRNA基因，螺旋体16S rRNA和23S rRNA基因进行扩增，对阳性产物进行克隆测序，测序获得的基因序列进行系统发育分析来确定病原的基因型。结果显示，从蜱样本中检测出5种蜱传细菌病原，包括敬信立克次体(Candidatus Rickettsia jingxinensis)、中央无形体(Anaplasma centrale)、扁平无形体(Anaplasma platys)、米氏埃立克体(Ehrlichia minasensis)和宫本疏螺旋体(Borrelia miyamotoi)。其中敬信立克次体、米氏埃立克体和宫本疏螺旋体阳性率均为2.16%,扁平无形体阳性率为0.30%,中央无形体阳性率为5.79%。本研究利用物种鉴定与分子生物学相结合的方法，从分子流行病学角度解析这些病原体在蜱体内存在及流行情况，并且在湖南省蜱中检测到扁平无形体和宫本疏螺旋体两种人兽共患病原体，可为湖南省的虫媒疾病的研究和防控提供数据参考。

关键词(KeyWords)： 蜱传病原;立克次体;无形体;埃立克体;螺旋体

Abstract:

Keywords:

基金项目(Foundation): 广东省珠江人才计划资助项目(2019CX01N111);; 广东省佛山市科技创新资助项目(2020001000151);; 国家寄生虫资源中心资助项目(NPRC-2019-194-30)

作者(Author): 丁立,张力,刘全

DOI: 10.16303/j.cnki.1005-4545.2023.12.13

参考文献(References)：

• [1] HOPLA C E,DURDEN L A,KEIRANS J E.Ectoparasites and classification[J].Rev Sci Tech,1994,13(4):985-1017.
• [2] JONGEJAN F,UILENBERG G.The global importance of ticks[J].Parasitology,2004,129(Suppl):S3-S14.
• [3] PLATONOV A E,KARAN L S,KOLYASNIKOVA N M,et al.Humans infected with relapsing fever spirochete Borrelia miyamotoi,Russia[J].Emerg Infect Dis,2011,17(10):1816-1823.
• [4] JAHFARI S,HOFHUIS A,FONVILLE M,et al.Molecular detection of tick-borne pathogens in humans with tick bites and rrythema migrans,in the Netherlands[J].PLoS Negl Trop Dis,2016,10(10):e5042.
• [5] HANSFORD K M,FONVILLE M,JAHFARI S,et al.Borrelia miyamotoi in host-seeking Ixodes ricinus ticks in England[J].Epidemiol Infect,2015,143(5):1079-1087.
• [6] KUBIAK K,SZCZOTKO M,DMITRYJUK M.Borrelia miyamotoi-an emerging tuman tick-borne pathogen in Europe[J].Microorganisms,2021,9(1):103-106.
• [7] TABARA K,ARAI S,KAWABUCHI T,et al.Molecular survey of Babesia microti,Ehrlichia species and Candidatus neoehrlichia mikurensis in wild rodents from Shimane Prefecture,Japan[J].Microbiol Immunol,2007,51(4):359-367.
• [8] PAROLA P,PADDOCK C D,SOCOLOVSCHI C,et al.Update on tick-borne rickettsioses around the world:A geographic approach[J].Clin Microbiol Rev,2013,26(4):657-702.
• [9] LIU H,LI Q,ZHANG X,et al.Characterization of rickettsiae in ticks in northeastern China[J].Parasites Vectors,2016,9(1):498.
• [10] DONG X,CHEN X P,LIU N,et al.Co-circulation of multiple species of Rickettsiales bacteria in one single species of hard ticks in Shenyang,China[J].Ticks Tick Borne Dis,2014,5(6):727-733.
• [11] LU M,TIAN J H,YU B,et al.Extensive diversity of rickettsiales bacteria in ticks from Wuhan,China[J].Ticks Tick Borne Dis,2017,8(4):574-580.
• [12] LIU H,LIANG X,WANG H,et al.Molecular evidence of the spotted fever group Rickettsiae in ticks from Yunnan Province,Southwest China[J].Exp Appl Acarol,2020,80(3):339-348.
• [13] GUO W P,WANG Y H,LU Q,et al.Molecular detection of spotted fever group rickettsiae in hard ticks,northern China[J].Transbound Emerg Dis,2019,66(4):1587-1596.
• [14] KAMANI J,SCHAER J,UMAR A G,et al.Molecular detection and genetic characterization of Anaplasma marginale and Anaplasma platys in cattle in Nigeria[J].Ticks Tick Borne Dis,2022,13(4):101955.
• [15] AUBRY P,GEALE D W.A review of bovine anaplasmosis[J].Transbound Emerg Dis,2011,58(1):1-30.
• [16] GUO W P,ZHANG B,WANG Y H,et al.Molecular identification and characterization of Anaplasma capra and Anaplasma platys-like in Rhipicephalus microplus in Ankang,Northwest China[J].BMC Infect Dis,2019,19(1):434.
• [17] WEI W,LI J,WANG Y W,et al.Anaplasma platys-like infection in goats,Beijing,China[J].Vector Borne Zoonotic Dis,2020,20(10):755-762.
• [18] LI Y,CHEN Z,LIU Z,et al.Molecular survey of anaplasma and ehrlichia of Red Deer and Sika Deer in Gansu,China in 2013[J].Transbound Emerg Dis,2016,63(6):e228-e236.
• [19] REMESAR S,PRIETO A,GARCIA-DIOS D,et al.Diversity of Anaplasma species and importance of mixed infections in roe deer from Spain[J].Transbound Emerg Dis,2022,69(4):e374-e385.
• [20] BEN S M,BELKAHIA H,EL M N,et al.Anaplasma platys-like strains in ruminants from Tunisia[J].Infect Genet Evol,2017,49:226-233.
• [21] CABEZAS-CRUZ A,ZWEYGARTH E,VANCOVA M,et al.Ehrlichia minasensis sp.nov.,isolated from the tick Rhipicephalus microplus[J].Int J Syst Evol Microbiol,2016,66(3):1426-1430.
• [22] CABEZAS-CRUZ A,ZWEYGARTH E,AGUIAR D M.Ehrlichia minasensis,an old demon with a new name[J].Ticks Tick Borne Dis,2019,10(4):828-829.
• [23] AGUIAR D M,ZILIANI T F,ZHANG X,et al.A novel Ehrlichia genotype strain distinguished by the TRP36 gene naturally infects cattle in Brazil and causes clinical manifestations associated with ehrlichiosis[J].Ticks Tick Borne Dis,2014,5(5):537-544.
• [24] GAJADHAR A A,LOBANOV V,SCANDRETT W B,et al.A novel Ehrlichia genotype detected in naturally infected cattle in North America[J].Vet Parasitol,2010,173(3/4):324-329.
• [25] MOURA D A D,PESSOA A J J,NAKAZATO L,et al.Isolation and characterization of a novel pathogenic strain of Ehrlichia minasensis[J].Microorganisms,2019,7(11):709-711.
• [26] CRUZ A C,ZWEYGARTH E,RIBEIRO M F,et al.New species of Ehrlichia isolated from Rhipicephalus (Boophilus) microplus shows an ortholog of the E.canis major immunogenic glycoprotein gp36 with a new sequence of tandem repeats[J].Parasit Vectors,2012,5:291.
• [27] LI J,LIU X,MU J,et al.Emergence of a novel Ehrlichia minasensis strain,harboring the major immunogenic glycoprotein trp36 with unique tandem repeat and C-terminal region sequences,in haemaphysalis hystricis ticks removed from free-ranging sheep in Hainan Province,China[J].Microorganisms,2019,7(9):369.
• [28] JIANG B G,JIA N,JIANG J F,et al.Borrelia miyamotoi infections in humans and Ticks,Northeastern China[J].Emerg Infect Dis,2018,24(2):236-241.
• [29] GAO Y,LYU X L,HAN S Z,et al.First detection of Borrelia miyamotoi infections in ticks and humans from the northeast of Inner Mongolia,China[J].Acta Trop,2021,217:105857.
• [30] FUKUNAGA M,TAKAHASHI Y,TSURUTA Y,et al.Genetic and phenotypic analysis of Borrelia miyamotoi sp.nov.,isolated from the ixodid tick Ixodes persulcatus,the vector for Lyme disease in Japan[J].Int J Syst Bacteriol,1995,45(4):804-810.
• [31] GAOWA,WULANTUYA,SATO K,et al.Surveillance of Borrelia miyamotoi-carrying ticks and genomic analysis of isolates in Inner Mongolia,China[J].Parasit Vectors,2021,14(1):368.
• [32] ZHANG X A,TIAN F,LI Y,et al.Molecular detection and identification of relapsing fever Borrelia in ticks and wild small mammals in China[J].Emerg Microbes Infect,2022,11(1):2632-2635.