乳房链球菌多位点序列分型和遗传进化特征分析

doi:10.16431/j.cnki.1671-7236.2021.05.004

中国畜牧兽医 ›› 2021, Vol. 48 ›› Issue (5): 1535-1543.doi: 10.16431/j.cnki.1671-7236.2021.05.004

乳房链球菌多位点序列分型和遗传进化特征分析

扶绍东¹, 仇亚伟¹, 温晨¹, 张毅豪¹, 陈伟², 罗振华³, 张金秋⁴, 苗晋锋¹

1. 南京农业大学动物医学院, 南京 210095;
2. 塔里木大学动物科学学院, 新疆兵团塔里木动物疫病诊断与防控工程实验室, 阿拉尔 843300;
3. 英国克兰菲尔德大学水、能源&环境学院, 贝德福德郡 MK430AL;
4. 江苏省农业科学院动物免疫工程研究所, 南京 210014

收稿日期:2020-09-03 出版日期:2021-05-20 发布日期:2021-05-20
通讯作者: 张金秋, 苗晋锋 E-mail:jqzh03@126.com;mjf171647@126.com
作者简介:扶绍东(1997-),男,安徽广德人,硕士生,研究方向:感染与动物健康调控,E-mail:fsd19972019@163.com
基金资助:
国家重点研发计划政府间专项（2018YFE0102200）；南京农业大学-塔里木大学联合基金项目（KYLH201904）；新疆兵团重点领域科技攻关计划（2020AAB025）

Analysis of Multilocus Sequence Typing and Genetic Evolution of Streptococcus uberis

FU Shaodong¹, QIU Yawei¹, WEN Chen¹, ZHANG Yihao¹, CHEN Wei², LUO Zhenhua³, ZHANG Jinqiu⁴, MIAO Jinfeng¹

1. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China;
2. Engineering Laboratory of Tarim Animal Diseases Diagnosis and Control, Xinjiang Production & Construction Crops, College of Animal Science, Tarim University, Tarim 843300, China;
3. School of Water, Energy & Environment, Cranfield University, Bedfordshire MK430AL, United Kingdom;
4. Institute of Veterinary Immunology & Engineering, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China

Received:2020-09-03 Online:2021-05-20 Published:2021-05-20

摘要/Abstract

摘要： 为探究中国不同地区来源的乳房链球菌（Streptococcus uberis，S.uberis）之间的多样性和遗传进化关系，本研究以部分地区分离到的38株乳房链球菌为研究对象，通过对其最小抑菌浓度（minimun inhibitory concentration，MIC）测定和全基因组框架测序，获取38株菌耐药性、耐药基因及毒力基因等相关信息；通过多位点序列分型技术（multilocus sequence typing，MLST）分析了7个管家基因的等位基因谱、序列型（sequence type，ST）和克隆复合体（clonal complexes，CCs），并构建系统进化树。MIC试验共采用15种抗生素，除头孢噻呋和氟苯尼考外，38株乳房链球菌对其中13种抗生素具有不同程度耐药，且江苏地区的耐药性明显强于新疆地区；耐药和毒力基因比对后发现，仅24株菌携带耐药基因，剩余14株则不携带耐药基因且属新疆地区分离株，与MIC试验结果相符；15种毒力基因中，除cfu、hasA/B和lbp外，余下12种毒力基因的携带率均高达100%。MLST结果表明，38株乳房链球菌共属于17个ST型，其中有15个ST型从未报道，仅supan01 1株菌属于ST-5 CCs。进化树结果揭示，38株分离株之间的亲缘性较远，分布呈地区依赖性，且中国部分地区当前流行的乳房链球菌与西方国家流行的菌株差异较大。本研究丰富了乳房链球菌的MLST分型数据库，为了解中国乳房链球菌遗传特性和分布特点提供了参考依据。

关键词: 乳房链球菌; 多位点序列分型(MLST); 管家基因

Abstract: The purpose of this study was to explore the diversity and genetic evolution of Streptococcus uberis (S.uberis) from different regions in China.In this study,38 strains of S.uberis isolated from some regions of China were used as the research objects.Through the determination of their minimum inhibitory concentration (MIC) and the sequencing of the whole genome frame,the related information of drug resistance,drug resistance gene and virulence gene of 38 strains were obtained.The alleles,sequence type (ST) and clone complexes (CCs) of 7 housekeeping genes were analyzed by multilocus sequence typing technique (MLST),and their evolutionary tree was established.A total of 15 antibiotics were used in MIC test,it was found that 38 strains of S.uberis were resistant to 13 antibiotics except for cefotaxime and florfenicol,and the drug resistance in Jiangsu province was significantly stronger than that in Xinjiang province.After comparison of resistance and virulence genes,only 24 strains were found to carry drug resistance genes,and the remaining 14 strains did not carry drug resistance genes and belonged to Xinjiang isolates,which was consistent with the results of MIC test.Among the 15 virulence genes,except for cfu,hasA/B and lbp,the carrying rate of the remaining 12 virulence genes was as high as 100%.Based on the MLST's results,38 strains of S.uberis belonged to 17 ST types,of which 15 ST types were not reported,and only 1 strain of supan01 belonged to ST-5 CCs.The phylogenetic tree results showed that the 38 isolates largely unrelated,and their distribution was region-dependent.Moreover,the phylogenetic tree results suggested that the S.uberis strains prevalent in some parts of China were largely different from those prevalent in Western countries.This study contributed to the MLST database of S.uberis and provided a reference basis for understanding of the genetic and distribution characteristics of S.uberis in China.

Key words: Streptococcus uberis; multilocus sequence typing (MLST); housekeeping gene

中图分类号:

S852.61

扶绍东, 仇亚伟, 温晨, 张毅豪, 陈伟, 罗振华, 张金秋, 苗晋锋. 乳房链球菌多位点序列分型和遗传进化特征分析[J]. 中国畜牧兽医, 2021, 48(5): 1535-1543.

FU Shaodong, QIU Yawei, WEN Chen, ZHANG Yihao, CHEN Wei, LUO Zhenhua, ZHANG Jinqiu, MIAO Jinfeng. Analysis of Multilocus Sequence Typing and Genetic Evolution of Streptococcus uberis[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(5): 1535-1543.

参考文献

[1] VITALI A,FELICI A,LEES A M,et al.Heat load increases the risk of clinical mastitis in dairy cattle[J].Journal of Dairy Science,2020,103(9):8378-8387.
[2] CÔTÉ-GRAVEL J,MALOUIN F.Symposium review:Features of Staphylococcus aureus mastitis pathogenesis that guide vaccine development strategies[J].Journal of Dairy Science,2019,102(5):4727-4740.
[3] BURVENICH C,VAN MERRIS V R,MEHRZAD J,et al.Severity of E.coli mastitis is mainly determined by cow factors[J].Veterinary Research,2003,34(5):521-564.
[4] KLAAS I C,ZADOKS R N.An update on environmental mastitis:Challenging perceptions[J].Transboundary and Emerging Diseases,2018,65(1):166-185.
[5] LI B,XI P,WANG Z,et al.PI3K/Akt/mTOR signaling pathway participates in Streptococcus uberis-induced inflammation in mammary epithelial cells in concert with the classical TLRs/NF-κB pathway[J].Veterinary Microbiology,2018,227:103-111.
[6] ABUREEMA S,DEIGHTON M,MANTRI N.A novel subtraction diversity array distinguishes between clinical and non-clinical Streptococcus uberis and identifies potential virulence determinants[J].Veterinary Microbiology,2019,237:108385.
[7] 汪智,王涓,吴清平,等.食源性空肠弯曲菌的分子分型及其遗传多样性分析[J].现代食品科技,2020,36(3):133-139. WANG Z,WANG J,WU Q P,et al.Molecular typing of foodborne Campylobacter jejuni and analysis of its genetic diversity[J].Modern Food Science and Technology, 2020,36(3):133-139.(in Chinese)
[8] LIAO F,MO Z,GU W,et al.A comparative genomic analysis between methicillin-resistant Staphylococcus aureus strains of hospital acquired and community infections in Yunnan province of China[J].BMC Infectious Diseases,2020,20(1):137.
[9] VLEZ J R,CAMERON M,RODRÍGUEZ-LECOMPTE J C,et al.Whole-genome sequence analysis of antimicrobial resistance genes in Streptococcus uberis and Streptococcus dysgalactiae isolates from canadian dairy herds[J].Frontiers in Veterinary Science,2017,4:63.
[10] DAVIES P L,LEIGH J A,BRADLEY A J,et al.Molecular epidemiology of Streptococcus uberis clinical mastitis in dairy herds:Strain heterogeneity and transmission[J].Journal of Clinical Microbiology,2016,54(1):68-74.
[11] 王丽君,陈伟,张利莉.奶牛乳房炎相关链球菌PCR检测方法的建立[J].畜牧与兽医,2010,42(10):7-9. WANG L J,CHEN W,ZHANG L L,et al.Establishment of PCR for Streptococci involved in bovine mastitis[J].Animal Husbandry & Veterinary Medicine,2010,42(10):7-9.(in Chinese)
[12] MITRA S D,SHOME B R,MANI B,et al.Streptococcus uberis ST439 and ST475 induce differential inflammatory responses in a mouse intrama-mmary infection model[J].Gene,2016,585(2):247-255.
[13] COFFEY T J,PULLINGER G D,URWIN R,et al.First insights into the evolution of Streptococcus uberis:A multilocus sequence typing scheme that enables investigation of its population biology[J].Applied and Environmental Microbiology,2006,72(2):1420-1428.
[14] KPPELI N,MORACH M,ZURFLUH K,et al.Sequence types and antimicrobial resistance profiles of Streptococcus uberis isolated from bovine mastitis[J].Frontiers in Veterinary Science,2019,6:234.
[15] PETINAKI E,GUE RIN-FAUBLE E V,PICHEREAU V,et al.Lincomycin resistance gene lnu(D) in Streptococcus uberis[J].Antimicrobial Agents and Chemotherapy,2008,52(2):626-630.
[16] KHAN I U,HASSAN A A,ABDULMAWJOOD A,et al.Identification and epidemiological characterization of Streptococcus uberis isolated from bovine mastitis using conventional and molecular methods[J].Journal of Veterinary Science,2003,4(3):213-224.
[17] WARD P N,HOLDEN M T,LEIGH J A,et al.Evidence for niche adaptation in the genome of the bovine pathogen Streptococcus uberis[J].BMC Genomics,2009,10:54.
[18] FIELD T R,WARD P N,PEDERSEN L H,et al.The hyaluronic acid capsule of Streptococcus uberis is not required for the development of infection and clinical mastitis[J].Infection and Immunity,2003,71(1):132-139.
[19] REINOSO E B,LASAGNO M C,DIESER S A,et al.Distribution of virulence-associated genes in Streptococcus uberis isolated from bovine mastitis[J].FEMS Micro-biology Letters,2011;318(2):183-188.
[20] PATEL D,ALMEIDA R A,DUNLAP J R,et al.Bovine lactoferrin serves as a molecular bridge for internalization of Streptococcus uberis into bovine mammary epithelial cells[J].Veterinary Microbiology,2009,137(3-4):297-301.
[21] 张少敏,徐建国.多位点序列分型及其应用[J].疾病监测,2008,23(10):648-650. ZHANG S M,XU J G.Multilocus sequence typing and its application[J].Disease Surveillance,2008,23(10):648-650.(in Chinese)
[22] 邬琴,张星星,顾晓晓,等.牛源多杀性巴氏杆菌血清分型及毒力相关基因的检测研究[J].中国畜牧兽医,2020,47(6):1910-1920. WU Q,ZHANG X X,GU X X,et al.Study on serotyping and detection of virulence-associated gene of Pasteurella multocida isolated from bovine[J].China Animal Husbandry & Veterinary Medicine,2020,47(6):1910-1920.(in Chinese)
[23] PULLINGER G D,LÓPEZ-BENAVIDES M,COFFEY T J,et al.Application of Streptococcus uberis multilocus sequence typing:Analysis of the population structure detected among environmental and bovine isolates from New Zealand and the United Kingdom[J].Applied and Environmental Microbiology,2006,72(2):1429-1436.
[24] WALD R,BAUMGARTNER M,GUTSCHIREITER J,et al.Comparison of the population structure of Streptococcus uberis mastitis isolates from Austrian small-scale dairy farms and a Slovakian large-scale farm[J].Journal of Dairy Science,2020,103(2):1820-1830.

乳房链球菌多位点序列分型和遗传进化特征分析

Analysis of Multilocus Sequence Typing and Genetic Evolution of Streptococcus uberis

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