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从巴西奶牛场采集的牛奶中分离出的金黄色葡萄球菌荚膜多糖和生物膜形成的基因型和表型检测

Genotypic and phenotypic detection of capsular polysaccharide and biofilm formation in Staphylococcus aureus isolated from bovine milk collected from Brazilian dairy farms.

作者信息

Salimena Alessandra P S, Lange Carla C, Camussone Cecilia, Signorini Marcelo, Calvinho Luis F, Brito Maria A V P, Borges Cristiano A V, Guimarães Alessandro S, Ribeiro João B, Mendonça Letícia C, Piccoli Roberta H

机构信息

Embrapa Dairy Cattle Research Center, Rua Eugênio do Nascimento 610, 36038-330, Juiz de Fora, Minas Gerais, Brazil.

Food Science Department, Federal University of Lavras, PO Box 3037, 37200-000, Lavras, Minas Gerais, Brazil.

出版信息

Vet Res Commun. 2016 Dec;40(3-4):97-106. doi: 10.1007/s11259-016-9658-5. Epub 2016 Jun 2.

Abstract

Staphylococcus aureus is a pathogen that frequently causes mastitis in bovine herds worldwide. This pathogen produces several virulence factors, including cell-associated adhesins, toxic and cytolytic exoproteins, and capsular polysaccharides. The aim of the present study was to test for the presence of genes involved in capsular polysaccharide production and biofilm formation in S. aureus isolated from bovine mastitis samples collected from 119 dairy herds located in three different Brazilian regions, as well as to assay the production of capsular polysaccharides and biofilm, in vitro. The detection of the cap, icaAD, and bap genes was performed using PCR. The detection and quantification of capsular polysaccharide production was performed using ELISA assays. The ability of the isolates to form a biofilm was examined using the polystyrene surface of microtiter plates. All 159 S. aureus isolates investigated harboured the cap gene: 80 % carried the cap5 gene and 20 % carried the cap8 gene. Sixty-nine percent of the isolates expressed capsular polysaccharide (CP) in vitro, 58 % expressed CP5 and 11 % expressed CP8. All of the isolates harboured the icaA and icaD genes, and 95.6 % of the isolates carried the bap gene. Of the 159 isolates analysed, 97.5 % were biofilm producers. A significant association between the capsular genotype and phenotype and the amount of biofilm formation was detected: cap5/CP5 isolates tended to form more biofilm and to produce a thinner CP layer than cap8/CP8 isolates. The results indicate a high potential for pathogenicity among S. aureus isolated from bovine milk collected from three different regions in Brazil.

摘要

金黄色葡萄球菌是一种在全球范围内经常导致牛群患乳腺炎的病原体。这种病原体产生多种毒力因子,包括细胞相关黏附素、毒性和细胞溶解性外蛋白以及荚膜多糖。本研究的目的是检测从巴西三个不同地区的119个奶牛场采集的乳腺炎样本中分离出的金黄色葡萄球菌中参与荚膜多糖产生和生物膜形成的基因的存在情况,并在体外测定荚膜多糖和生物膜的产生。使用PCR检测cap、icaAD和bap基因。使用ELISA检测法检测和定量荚膜多糖的产生。使用微量滴定板的聚苯乙烯表面检测分离株形成生物膜的能力。所研究的159株金黄色葡萄球菌均携带cap基因:80%携带cap5基因,20%携带cap8基因。69%的分离株在体外表达荚膜多糖(CP),58%表达CP5,11%表达CP8。所有分离株均携带icaA和icaD基因,95.6%的分离株携带bap基因。在分析的159株分离株中,97.5%是生物膜产生菌。检测到荚膜基因型和表型与生物膜形成量之间存在显著关联:cap5/CP5分离株比cap8/CP8分离株倾向于形成更多生物膜并产生更薄的CP层。结果表明,从巴西三个不同地区采集的牛奶中分离出的金黄色葡萄球菌具有很高的致病潜力。

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