广西罗非鱼无乳链球菌的分离鉴定、毒力基因及耐药性研究

doi:10.16431/j.cnki.1671-7236.2021.07.037

中国畜牧兽医 ›› 2021, Vol. 48 ›› Issue (7): 2607-2616.doi: 10.16431/j.cnki.1671-7236.2021.07.037

广西罗非鱼无乳链球菌的分离鉴定、毒力基因及耐药性研究

郭长明¹, 陈怀君^1,2, 袁橙¹, 罗福广³, 黄德生⁴, 贾国华⁵, 袁圣¹, 朱善元¹

1. 江苏农牧科技职业学院, 江苏省兽用生物制药高技术研究重点实验室, 泰州 225300;
2. 广西大学动物科学技术学院, 南宁 530004;
3. 柳州市渔业技术推广站, 柳州 545006;
4. 合浦县水产技术推广站, 北海 536100;
5. 湛江恒兴养殖技术服务有限公司, 湛江 524000

收稿日期:2021-01-18 出版日期:2021-07-20 发布日期:2021-07-15
通讯作者: 朱善元 E-mail:jstzzsy@126.com
作者简介:郭长明(1984-),男,山东临朐人,博士,副教授,研究方向:兽医微生物学与免疫学,E-mail:gcmscience@126.com
基金资助:
江苏高校"青蓝工程"项目（苏教师函[2021]11号）；江苏省高等学校自然科学研究面上项目（19KJB230005）；泰州市科技支撑计划（农业）项目（TN201916）；泰州市311高层次人才培养工程（[2019]21号）；江苏农牧科技职业学院科研项目（NSF20190202）

Isolation,Identification,Virulence Genes and Antibiotic Resistance Analysis of Streptococcus agalactiae from Tilapia in Guangxi

GUO Changming¹, CHEN Huaijun^1,2, YUAN Cheng¹, LUO Fuguang³, HUANG Desheng⁴, JIA Guohua⁵, YUAN Sheng¹, ZHU Shanyuan¹

1. Jiangsu Key Laboratory for High-Tech Research and Development of Veterinary Biopharmaceuticals, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, China;
2. College of Animal Science and Technology, Guangxi University, Nanning 530004, China;
3. Liuzhou Fishery Technology Extension Station, Liuzhou 545006, China;
4. Hepu Aquatic Technology Extension Station, Beihai 536100, China;
5. Zhanjiang Hengxing Breeding Technology Service Co., Ltd., Zhanjiang 524000, China

Received:2021-01-18 Online:2021-07-20 Published:2021-07-15

摘要/Abstract

摘要： 本研究旨在明确中国罗非鱼主养区广西各地罗非鱼无乳链球菌（Streptococcus agalactiae）分离株的血清型分布、毒力基因携带情况和耐药情况，为罗非鱼无乳链球菌病的防控奠定基础。2018-2019年从广西柳州、钦州、南宁、北海等地患无乳链球菌病罗非鱼体内分离了47株无乳链球菌，并对各分离株的血清型、毒力基因分布和耐药情况进行检测和分析。血清型检测结果表明，47株无乳链球菌血清型高度一致，均为Ⅰa血清型。对4种毒力基因检测结果表明，47株无乳链球菌临床分离株cylE、sodA、gapC毒力基因的检出率均为100%，而scpB基因仅在人源参考菌株2603V/R中检出，在所有鱼源分离株中未检出。对11类（31种）常见抗生素的药敏试验结果表明，临床分离株对磺胺异噁唑、新霉素、庆大霉素、卡那霉素的耐药率达到90%以上，对氧氟沙星、左氧氟沙星、氨苄青霉素、阿莫西林、头孢克洛、头孢曲松、头孢哌酮、头孢拉定、土霉素、强力霉素的敏感性均为100%。多重耐药检测结果表明，5重以上耐药菌株占93.62%，其中9重以上耐药菌株为19.15%，且均分离自柳州地区。结果表明，广西地区罗非鱼源无乳链球菌血清型单一，均为Ⅰa血清型，且均携带多种毒力基因，多重耐药现象严重。

关键词: 无乳链球菌; 罗非鱼; 血清型; 毒力基因; 耐药性

Abstract: The purpose of this study was to determine the serotype distribution,virulence gene carrying and drug resistance of Streptococcus agalactiae (S.agalactiae) isolated from tilapia in Guangxi,China,and laid a foundation for the prevention and control of S.agalactiae.47 strains of S.agalactiae were isolated from tilapia farms in Liuzhou,Qinzhou,Nanning and Beihai of Guangxi in 2018-2019,the serotype,distribution of virulence gene and drug resistance were detected and analyzed.The results of serotype test showed that 47 clinical isolates of S.agalactiae strains were Ⅰa serotype.The results of virulence gene detection showed that all clinical isolates of S.agalactiae carried cylE,sodA and gapC genes.The scpB gene was only detected in human reference strain 2603V/R,but not detected in all isolates of piscine S.agalactiae.The results of drug sensitivity test showed that the drug resistance rate of all the piscine isolates to sulfamethoxazole,neomycin,gentamicin and kanamycin was more than 90%,the drug sensitivity of the 47 piscineisolates to of loxacin,levofloxacin,ampicillin,amoxicillin,cefaclor,ceftriaxone,cefoperazone,cefradine,terramycin and doxycycline was 100%.All piscine strains had multiple drug resistance,and 93.62% of the 47 strains were resistanted to more than 5 kinds of drugs.Among all the piscine strains,the isolates resistanted to 9 drugs were accounted for 19.15% and isolated from Liuzhou.The results showed that the serotype of S.agalactiae isolated from tilapia in Guangxi was Ⅰa serotype.All 47 piscine isolates of S.agalactiae carried multiple virulence genes,and the phenomenon of multiple drug resistance was serious.

Key words: Streptococcus agalactiae; tilapia; serotype; virulence gene; antibiotic resistance

中图分类号:

S941.42

郭长明, 陈怀君, 袁橙, 罗福广, 黄德生, 贾国华, 袁圣, 朱善元. 广西罗非鱼无乳链球菌的分离鉴定、毒力基因及耐药性研究[J]. 中国畜牧兽医, 2021, 48(7): 2607-2616.

GUO Changming, CHEN Huaijun, YUAN Cheng, LUO Fuguang, HUANG Desheng, JIA Guohua, YUAN Sheng, ZHU Shanyuan. Isolation,Identification,Virulence Genes and Antibiotic Resistance Analysis of Streptococcus agalactiae from Tilapia in Guangxi[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(7): 2607-2616.

参考文献

[1] 李新.不同植物脂肪源对吉富罗非鱼生长、组织脂肪酸和生理生化指标的影响[D].武汉:华中农业大学,2016. LI X.Effect of dietary vegetable oils on growth,tissue fatty acid composition and physiological and biochemical index of genetically improved farmed tilapia (Oreochromis niloticus)[D].Wuhan:Huazhong Agricultural University,2016.(in Chinese)
[2] 张德锋,袁伟,可小丽,等.中国罗非鱼主养区无乳链球菌的分子流行特征及其传播方式[J].中国水产科学,2017,24(3):606-614. ZHANG D F,YUAN W,KE X L,et al.Molecular characteristics and transmission of Streptococcus agalactiae in a major tilapia culturing area of China[J].Journal of Fishery Sciences of China,2017,24(3):606-614.(in Chinese)
[3] 梁静真,黄立春,韦慕兰,等.广西罗非鱼源无乳链球菌耐药性及其四环素耐药基因检测[J].南方农业学报,2018,49(10):2077-2086. LIANG J Z,HUANG L C,WEI M L,et al.Detection of antibiotics resistance and tetracycline resistance genes in Streptococcus agalactiae from tilapia in Guangxi[J].Journal of Southern Agriculture,2018,49(10):2077-2086.(in Chinese)
[4] 衣萌萌,王淼,李忠徽,等.无乳链球菌感染对吉富罗非鱼行为的影响及其机制[J].农业生物技术学报,2019,27(5):799-808. YI M M,WANG M,LI Z H,et al.The impacts and mechanism of Streptococcus agalactiae on behavior of Gift tilapia (Oreochromis niloticus),2019,27(5):799-808.(in Chinese)
[5] MHALU F S.Infection with Streptococcus agalactiae in a London hospital[J].Journal of Clinical Pathology,1976,29(4):309-312.
[6] LAKEW B T,FAYERA T,ALI Y M,et al.Risk factors for bovine mastitis with the isolation and identification of Streptococcus agalactiae from farms in and around Haramaya district,Eastern Ethiopia[J].Tropical Animal Health and Production,2019,51(6):1507-1513.
[7] VANDAMME P,DEVRIESE L A,POT B,et al.Streptococcus difficile is a nonhemolytic group B,type Ⅰb Streptococcus[J].International Journal of Systematic Bacteriology,1997,47(1):81-85.
[8] ZAMRI S M,AMAL M N,SITI Z A,et al.Control and prevention of Streptococcosis in cultured tilapia in Maaysia:A review[J].Pertanika Journal of Tropical Agricultural Science,2014,37(4):389-410.
[9] 黄木珍,黎炯,潘忠超,等.宝石鲈无乳链球菌的分离鉴定及耐药性分析[J].中国畜牧兽医,2014,41(10):247-251. HUANG M Z,LI J,PAN Z C,et al.Isolation,identification and drug susceptibility analysis of Streptococcus agalactiae in Scortumbarcoo[J].China Animal Husbandry & Veterinary Medicine,2014,41(10):247-251.(in Chinese)
[10] 黎源,王蓓,汪志文,等.罗非鱼无乳链球菌传播途径与逃避宿主免疫防御策略[J].广东农业科学,2017,44(7):132-140. LI Y,WANG B,WANG Z W,et al.Transmission routes of Streptococcus agalactiae and the strategy of immune evasion in tilapia[J].Guangdong Agricultural Sciences,2017,44(7):132-140.(in Chinese)
[11] 李莉萍,王瑞,黄婷,等.2007-2012年中国罗非鱼无乳链球菌流行菌株血清型分析[J].大连海洋大学学报,2014,9(5):469-475. LI L P,WANG R,HUANG T,et al.Serotype of Streptococcus agalactiae isolated from Nile tilapia Oreochromis niloticus in China from 2007 to 2012[J].Journal of Dalian Ocean University,2014,9(5):469-475.(in Chinese)
[12] LI L P,WANG R,LIANG W W,et al.Rare serotype occumrence and PFGE genotypic diversity of Streptococcus agalactiae isolated from tilapia in China[J].Veterinary Microbiology,2013,167(3-4):719-724.
[13] 胡安东,张明洋,张飘,等.鲤源嗜水气单胞菌分离鉴定、耐药性及毒力基因检测[J].中国畜牧兽医,2019,46(7):2167-2175. HU A D,ZHANG M Y,ZHANG P,et al.Isolation,identification,drug resistance and virulence genes detection of Aeromonas hydrophila from carp[J].China Animal Husbandry & Veterinary Medicine,2019,46(7):2167-2175.(in Chinese)
[14] MOXON E R,KROLL J S.The role of bacterial polysaccharide capsules as virulence factors[J].Current Topics in Microbiology and Immunology,1990,150:65-85.
[15] 祝璟琳,杨弘.鱼源无乳链球菌致病机理研究进展[J].广东海洋大学学报,2013,33(6):92-96. ZHU J L,YANG H.Review on pathogenesis of Streptococcus agalactiae from fish[J].Journal of Guangdong Ocean University,2013,33(6):92-96.(in Chinese)
[16] BOHNSACK J F,TAKAHASHI S,HAMMITT L,et al.Genetic polymorphisms of group B Streptococcus scpB alter functional activity of a cell-associated peptidase that inactivates C5a[J].Infection and Immunity,2000,68(9):5018-5025.
[17] 邓永强,汪开毓.鱼类无乳链球菌病的研究进展[J].中国畜牧兽医,2016,43(9):2490-2495. DENG Y Q,WANG K Y.Research progress on fish Streptococcus agalactiae disease[J].China Animal Husbandry & Veterinary Medicine,2016,43(9):2490-2495.(in Chinese)
[18] BARKHAM T,ZADOKS R N,AZMAI M N A,et al.One hypervirulent clone,sequence type 283,accounts for a large proportion of invasive Streptococcus agalactiae isolated from humans and diseased tilapia in Southeast Asia[J].PLoS Neglected Tropical Diseases,2019,13(6):e0007421.
[19] MARTINEZ G,HAREL J,GOTTSCHALK M.Specific detection by PCR of Streptococcus agalactiae in milk[J].Canadian Journal of Veterinary Research,2001,65(1):68-72.
[20] BAKER G C,SMITH J J,COWAN D A.Review and re-analysis of domain-specific 16S primers[J].Journal of Microbiological Methods,2003,55(3):541-555.
[21] IMPERI M,PATARACCHIA M,ALFARONE G,et al.A multiplex PCR assay for the direct identification of the capsular type (Ⅰa to Ⅸ) of Streptococcus agalactiae[J].Journal of Microbiological Methods,2010,80(2):212-214.
[22] 刘广锦.无乳链球菌鱼源株的免疫原性蛋白筛选和比较基因组学分析[D].南京:南京农业大学,2013. LIU G J.Screening immunogenic proteins and comparative genomics analysis of piscine Streptococcus agalactiae[D].Nanjing:Nanjing Agricultural University,2013.(in Chinese)
[23] 王瑞.无乳链球菌跨宿主感染机制的比较基因组学研究[D].南宁:广西大学,2018. WANG R.Comparative genomic study of mechanisms of cross host infection in Streptococcus agalactiae[D].Nanning:Guangxi University,2018.(in Chinese)
[24] 陈福艳,黎建斌,欧阳贤华,等.鱼鸭混养池塘罗非鱼无乳链球菌病的病原分离鉴定及其防治[J].南方农业学报,2019,50(11):2583-2591. CHEN F Y,LI J B,OUYANG X H,et al.Isolation,identification and prevention of the pathogen ofStreptococcus agalactiae disease in tilapia of fish-duck polyculture pond[J].Journal of Southern Agriculture,2019,50(11):2583-2591.(in Chinese)
[25] DENG L S,LI Y J,GENG Y,et al.Molecular serotyping and antimicrobial susceptibility of Streptococcus agalactiae isolated from fish in China[J].Aquaculture,2019,510:84-89.
[26] ANUBHA S,CAROLIN K,LARA H,et al.The β-hemolysin and intracellular survival of Streptococcus agalactiae in human macrophages[J].PLoS One,2013,8(4):e60160.
[27] MARINA B,MARTA V,VANESSA M,et al.Streptococcus agalactiae GAPDH is a virulence-associated immunomodulatory protein[J].The Journal of Immunology,2007,178(3):1379-1387.
[28] 祝璟琳,季桓涛,肖炜,等.鱼腥草对无乳链球菌引起吉富罗非鱼肝脏损伤的修复作用[J].水产学报,2020,44(7):1187-1197. ZHU J L,JI H T,XIAO W,et al.Protective effects from Houttuyniacordata against Streptococcus agalactiae induced liver injury of Oreochromis niloticus GIFT strain[J].Journal of Fisheries of China,2020,44(7):1187-1197.(in Chinese)

广西罗非鱼无乳链球菌的分离鉴定、毒力基因及耐药性研究

Isolation,Identification,Virulence Genes and Antibiotic Resistance Analysis of Streptococcus agalactiae from Tilapia in Guangxi

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