一株羊源致病性屎肠球菌的分离鉴定及耐药性分析

doi:10.16431/j.cnki.1671-7236.2017.02.032

《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (2): 530-537.doi: 10.16431/j.cnki.1671-7236.2017.02.032

一株羊源致病性屎肠球菌的分离鉴定及耐药性分析

刘永安, 李学瑞, 周建华, 马丽娜, 刘永生

中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, 农业部草食动物疫病重点开放实验室, 兰州 730046

收稿日期:2016-08-03 出版日期:2017-02-20 发布日期:2017-02-25
通讯作者: 李学瑞, 刘永生 E-mail:lixuerui@caas.cn;liuyongsheng@caas.cn
作者简介:刘永安(1991-), 男, 山东济宁人, 硕士生, 研究方向:动物传染病病原学与流行病学, E-mail:457659155@qq.com
基金资助:
国家自然科学基金（31172335）

Isolation, Identification and Drug Resistance Analysis of Pathogenic Enterococcus feacium of Sheep

LIU Yong-an, LI Xue-rui, ZHOU Jian-hua, MA Li-na, LIU Yong-sheng

Key Laboratory of Grazing Animal Diseases of Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China

Received:2016-08-03 Online:2017-02-20 Published:2017-02-25

摘要/Abstract

摘要：

试验从甘肃肃南县某羊场死亡羊中分离出疑似某球菌菌株XCP，为进一步确定该致病菌株及其耐药性情况，进行了病原菌分离、革兰氏染色、生化试验、16S rRNA PCR扩增鉴定、小鼠致病力试验、药敏试验、毒力基因和耐药基因扩增试验研究。结果发现，该菌革兰氏染色阳性，高盐、蜜二糖、蔗糖等生化反应阳性，VP、马尿酸盐、阿拉伯糖等生化反应阴性；16S rRNA基因PCR扩增后，进行BLAST比对，结果与GenBank中的屎肠球菌16S rRNA基因序列同源性为100%；小鼠致病力试验发现，该菌株具有很强的致病性；该菌株对β-内酰胺类抗生素苯唑西林、青霉素G，喹诺酮类抗生素诺氟沙星、环丙沙星、左氟沙星，以及四环素等高度耐药，对红霉素、万古霉素、克林霉素等抗生素敏感；毒力基因Asal、cylA、acm和耐药基因TetM、ant（6）-Ⅰ、aac（6'）-aph（2"）、ermB扩增均为阳性。结果表明，该菌为屎肠球菌（Enterococcus faecium），具有较强的致病性和耐药性，可能与毒力基因和耐药基因的高阳性率有关。

关键词: 屎肠球菌; 分离鉴定; 耐药性; 毒力因子; 耐药基因

Abstract:

To further identify the pathogenic strains and analyze their antibiotic resistance, the methods of pathogen isolating, the methods of Gram staining, biochemical tests and 16S rRNA PCR amplification, virulence tests, drug sensitive tests, virulence genes and resistance genes PCR amplification were used. The results showed that the strain was Gram-positive and revealed positive after reacting with 6.5% NaCl, melibiose, sucrose and etc., while revealed negative reactions with VP, hippurate, arabinose and etc.. It showed 100% similarity with Enterococcus feacium 16S rRNA gene sequence in GenBank after PCR amplification of 16S rRNA gene and BLAST alignment. It was found severe pathogenicity after virulence tests in mice. The strain was highly resistant to oxacillin, penicillin G of β-lactam antibiotics and norfloxacin, ciprofloxacin, levofloxacin of quinolones antibiotics and tetracycline, while was sensitive to antibiotics of erythromycin, vancomycin and clindamycin. It revealed that virulence factor genes Asal, cylA, acm and resistance genes TetM, ant(6)-Ⅰ, aac(6')-aph (2"), ermB showed positive. The results showed that the bacteria was Enterococcus feacium, it had a strong pathogenicity and severe drug resistance, which might be related to the highly positive rates of virulence genes and drug resistance genes.

Key words: Enterococcus feacium; isolation and identification; drug resistance; virulence factor; resistance gene

中图分类号:

S852.61⁺1

刘永安, 李学瑞, 周建华, 马丽娜, 刘永生. 一株羊源致病性屎肠球菌的分离鉴定及耐药性分析[J]. 《中国畜牧兽医》, 2017, 44(2): 530-537.

LIU Yong-an, LI Xue-rui, ZHOU Jian-hua, MA Li-na, LIU Yong-sheng. Isolation, Identification and Drug Resistance Analysis of Pathogenic Enterococcus feacium of Sheep[J]. , 2017, 44(2): 530-537.

参考文献

[1] 刘佳,陈霞,赵爱兰,等.河南省某地区健康人源及动物源肠球菌种属分布及耐药性差异研究[J].中国畜牧兽医,2014,41(12):172-177.
[2] Arias C A, Murray B E. The rise of the Enterococcus: Beyond vancomycin resistance[J]. Nat Rev Microbiol, 2012, 10(4):266-278.
[3] Mutschler M, Trojan S, Defosse J M, et al. Severe sepsis caused by a linezolid-resistant Enterococcus faecium in a 10-year-old girl after multiple trauma[J]. International Journal of Infectious Diseases, 2013, 17 (6):e466-e467.
[4] Kang M,Xie Y, He C, et al. Molecular characteristics of vancomycin-resistant Enterococcus faecium from a tertiary care hospital in Chengdu, China: Molecular characteristics of VRE in China[J].Eur J Clin Microbiol Infect Dis,2014, 33(6):933-939.
[5] 阳苹,孙珊,张莉萍,等.重庆市1264株粪肠球菌和屎肠球菌耐药性监测[J].中国微生态学杂志,2013,25(9):1028-1031.
[6] 杨青,俞云松,倪语星,等.2010年中国CHINET肠球菌属细菌耐药性监测[J].中国感染与化疗杂志,2012,12(2):92-97.
[7] Spiliopoulou I,Anastassiou, E D,Petinaki E. Comment on:Intensive care unit dissemination of multiple clones of linezolid-resistant Enterococcus faecalis and Enterococcusfaecium[J]. J Antimicrob Chemother,2012, 67(12):3015-3016.
[8] Baele M, Baele P, Vaneechoutte M, et al. Application of tRNA intergenic spacer PCR for identification of Enterococcus species[J]. J Clin Microbiol, 2000, 38:4201-4207.
[9] 张芬芬,李学瑞,杨建设.屎肠球菌新序列型ST989菌株的分离鉴定及多重耐药性分析[J].中国畜牧兽医,2015,42(5):1274-1282.
[10] Leavis H, Top J, Shankar N, et al. Anovel putative enterococcal pathogenicity island linked to the esp virulence gene of Enterococcus faecium and associated with epidemicity[J]. J Bacteriol, 2004, 186(3):672-682.
[11] Sun H, Wang H, Xu Y, et al. Molecular characterization of vancomycin-resistant Enterococcus spp. clinical isolates recovered from hospitalized patients among several medical institutions in China[J]. Diagnostic Microbiology and Infectious Disease, 2012, 74(4):399-403.
[12] 周娟,金东,卢珊,等.青藏高原牦牛携带屎肠球菌及其毒力基因和耐药性检测[J].中国人兽共患病学报,2015,31(4):298-302.
[13] 李爽,张正.粪肠球菌与屎肠球菌药敏表型和耐药基因的比较[J].临床检验杂志,2005,23(3):174-176.
[14] 陈蓉,章小小,陈楠,等. 一例灰袋鼠致病性屎肠球菌的分离鉴定[J]. 野生动物学报,2015,36(4):443-446.
[15] 邓梅葵,孙迎,韩雯晴.细菌鉴定方法[J].生物医学进展,2014,35(2):84-88.
[16] Coburn P S, Baghdayan A S, Dolan G T, et al. An AraC-type transcriptional regulator encoded on the Enterococcus faecalis pathogenicity island contributes to pathogenesis and intracellular macrophage survival[J]. Infect Immun, 2008, 76(12):5668-5676.
[17] 郭秉娇,王熙楚,秦瑶,等.不同动物来源肠球菌的分离鉴定及毒力因子基因的检测[J].中国兽医科学,2013,43(10):1021-1027.
[18] Yu H B, Zhang Y L, Liu Y L, et al. Identification and characterization of putative virulence of genes and gene clusters in aeromonas hydrophila PPD134/91[J]. Appl Environ Microbiol, 2005, 71(8):4469-4477.
[19] Semedo T, Almeida S M, Martins P, et al. Comparative study using type strains and clinical and food isolates to examine hemolytic activity and occurrence of the cyl operon in Enterococci[J].J Clin Microbiol, 2003, 41(6):2569-2576.
[20] Creti R, Imperi M, Bertuccini L, et al. Survey for virulence determinants among Enterococcus faecalis isolated from different sources[J]. J Med Microbiol, 2004, 53(1):13-20.
[21] Elisa B M, Sergio S. Occurrence of virulence associated genes in clinical Enterococcus faecalis strains isolated in Londrina, Brazil[J]. J Med Microbiol, 2004, 53:1069-1073.
[22] Varhimo E, Varmanen P, Fallarero A, et al. Alpha- and β-casein components of host milk induce biofilm formation in the mastitis bacterium Streptococcus uberis[J]. Vet Microbiol, 2011, 149(3-4):381-389.
[23] 卢彩景,张秀平,陈陆,等.猪源屎肠球菌致病力与耐药性及生物被膜形成能力的相关性分析[J].中国预防兽医学报,2014,36(5):350-353.

一株羊源致病性屎肠球菌的分离鉴定及耐药性分析

Isolation, Identification and Drug Resistance Analysis of Pathogenic Enterococcus feacium of Sheep

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