Jørgensen H J, Mørk T, Caugant D A, Kearns A, Rørvik L M
Department of Food and Feed Hygiene, National Veterinary Institute, P.O. Box 8156 Dep., 0033 Oslo, Norway.
Appl Environ Microbiol. 2005 Dec;71(12):8352-61. doi: 10.1128/AEM.71.12.8352-8361.2005.
Strains of Staphylococcus aureus obtained from bovine (n = 117) and caprine (n = 114) bulk milk were characterized and compared with S. aureus strains from raw-milk products (n = 27), bovine mastitis specimens (n = 9), and human blood cultures (n = 39). All isolates were typed by pulsed-field gel electrophoresis (PFGE). In addition, subsets of isolates were characterized using multilocus sequence typing (MLST), multiplex PCR (m-PCR) for genes encoding nine of the staphylococcal enterotoxins (SE), and the cloverleaf method for penicillin resistance. A variety of genotypes were observed, and greater genetic diversity was found among bovine than caprine bulk milk isolates. Certain genotypes, with a wide geographic distribution, were common to bovine and caprine bulk milk and may represent ruminant-specialized S. aureus. Isolates with genotypes indistinguishable from those of strains from ruminant mastitis were frequently found in bulk milk, and strains with genotypes indistinguishable from those from bulk milk were observed in raw-milk products. This indicates that S. aureus from infected udders may contaminate bulk milk and, subsequently, raw-milk products. Human blood culture isolates were diverse and differed from isolates from other sources. Genotyping by PFGE, MLST, and m-PCR for SE genes largely corresponded. In general, isolates with indistinguishable PFGE banding patterns had the same SE gene profile and isolates with identical SE gene profiles were placed together in PFGE clusters. Phylogenetic analyses agreed with the division of MLST sequence types into clonal complexes, and isolates within the same clonal complex had the same SE gene profile. Furthermore, isolates within PFGE clusters generally belonged to the same clonal complex.
对从牛(n = 117)和羊(n = 114)的批量牛奶中分离得到的金黄色葡萄球菌菌株进行了特征分析,并与来自生鲜乳产品(n = 27)、牛乳腺炎标本(n = 9)和人类血培养物(n = 39)中的金黄色葡萄球菌菌株进行了比较。所有分离株均通过脉冲场凝胶电泳(PFGE)进行分型。此外,还使用多位点序列分型(MLST)、针对编码9种葡萄球菌肠毒素(SE)的基因的多重PCR(m-PCR)以及青霉素耐药性的三叶草叶法对部分分离株进行了特征分析。观察到了多种基因型,并且在牛的批量牛奶分离株中发现的遗传多样性比羊的更大。某些具有广泛地理分布的基因型在牛和羊的批量牛奶中都很常见,可能代表反刍动物特有的金黄色葡萄球菌。在批量牛奶中经常发现基因型与反刍动物乳腺炎菌株无法区分的分离株,并且在生鲜乳产品中也观察到了基因型与批量牛奶菌株无法区分的菌株。这表明来自感染乳房的金黄色葡萄球菌可能污染批量牛奶,进而污染生鲜乳产品。人类血培养分离株具有多样性,与其他来源的分离株不同。PFGE、MLST和SE基因的m-PCR分型结果在很大程度上是一致的。一般来说,PFGE条带模式无法区分的分离株具有相同的SE基因谱,而具有相同SE基因谱的分离株在PFGE聚类中被归为一类。系统发育分析与将MLST序列类型划分为克隆复合体的结果一致,并且同一克隆复合体内的分离株具有相同的SE基因谱。此外,PFGE聚类中的分离株通常属于同一克隆复合体。