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中国宁夏奶牛乳房炎分离金黄色葡萄球菌的药敏性、毒力基因及随机扩增多态性DNA分析

Antimicrobial susceptibility, virulence genes, and randomly amplified polymorphic DNA analysis of Staphylococcus aureus recovered from bovine mastitis in Ningxia, China.

作者信息

Wang Dong, Zhang Limei, Zhou Xuezhang, He Yulong, Yong Changfu, Shen Mingliang, Szenci Otto, Han Bo

机构信息

College of Life Science, Ningxia University, Yinchuan Ningxia 750021, China.

College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.

出版信息

J Dairy Sci. 2016 Dec;99(12):9560-9569. doi: 10.3168/jds.2016-11625. Epub 2016 Oct 19.

Abstract

Staphylococcus aureusis the leading pathogen involved inbovine mastitis, but knowledgeabout antimicrobial resistance, virulence factors, and genotypes of Staphylococcus aureus resulting in bovine mastitis in Ningxia, China, is limited. Therefore, antimicrobial susceptibility, virulence gene, and randomly amplified polymorphic DNA (RAPD) analyses of Staph. aureus were carried out. A total of 327 milk samples from cows with clinical and subclinical mastitis in 4 regions of Ningxia were used for the isolation and identification of pathogens according to phenotypic and molecular characteristics. Antimicrobial susceptibility against 22 antimicrobial agents was determined by disk diffusion. The presence of 8 virulence genes in Staph. aureus isolates was tested by PCR. Genotypes of isolates were investigated based on RAPD. Results showed that 35 isolates obtained from mastitis milk samples were identified as Staph. aureus. The isolates were resistant to sulfamethoxazole (100%), penicillin G (94.3%), ampicillin (94.3%), erythromycin (68.6%), azithromycin (68.6%), clindamycin (25.7%), amoxicillin (11.4%), and tetracycline (5.7%). All of the isolates contained one or more virulence genes with average (standard deviation) of 6.6±1.6. The most prevalent virulence genes were hlb (97.1%), followed by fnbpA, hla, coa (94.3% each), nuc (85.7%), fnbpB (80%), clfA (77.1%), and tsst-1 (40%). Nine different gene patterns were found and 3 of them were the dominant gene combinations (77.1%). Staphylococcus aureus isolates (n=35) were divided into 6 genotypes by RAPD tying, the genotypes III and VI were the most prevalent genotypes. There was greatvariation in genotypes of Staph. aureus isolates, not only among different farms, but also within the same herd in Ningxia province. The study showed a high incidence of Staph. aureus with genomic variation of resistance genes, which is matter of great concern in public and animal health in Ningxia province of China.

摘要

金黄色葡萄球菌是引起奶牛乳腺炎的主要病原菌,但在中国宁夏,关于导致奶牛乳腺炎的金黄色葡萄球菌的耐药性、毒力因子和基因型的了解有限。因此,对金黄色葡萄球菌进行了药敏试验、毒力基因和随机扩增多态性DNA(RAPD)分析。根据表型和分子特征,从宁夏4个地区患有临床和亚临床乳腺炎的奶牛中采集了327份乳样,用于病原菌的分离和鉴定。采用纸片扩散法测定了对22种抗菌药物的敏感性。通过PCR检测金黄色葡萄球菌分离株中8种毒力基因的存在情况。基于RAPD对分离株的基因型进行了研究。结果显示,从乳腺炎乳样中获得的35株分离株被鉴定为金黄色葡萄球菌。这些分离株对磺胺甲恶唑(100%)、青霉素G(94.3%)、氨苄西林(94.3%)、红霉素(68.6%)、阿奇霉素(68.6%)、克林霉素(25.7%)、阿莫西林(11.4%)和四环素(5.7%)耐药。所有分离株均含有一个或多个毒力基因,平均(标准差)为6.6±1.6。最常见的毒力基因是hlb(97.1%),其次是fnbpA、hla、coa(各94.3%)、nuc(85.7%)、fnbpB(80%)、clfA(77.1%)和tsst-1(40%)。发现了9种不同的基因模式,其中3种是主要的基因组合(77.1%)。通过RAPD分型将35株金黄色葡萄球菌分离株分为6种基因型,基因型III和VI是最常见的基因型。宁夏地区金黄色葡萄球菌分离株的基因型不仅在不同养殖场之间,而且在同一牛群内部都存在很大差异。该研究表明金黄色葡萄球菌的发生率较高且耐药基因存在基因组变异,这在中国宁夏的公共卫生和动物健康方面是一个令人高度关注的问题。

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