Institute for Biotechnology and Bioengineering/Center of Genomics and Biotechnology, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal.
Sci Total Environ. 2011 Dec 1;410-411:266-8. doi: 10.1016/j.scitotenv.2011.09.074. Epub 2011 Oct 20.
The aim of this study was to perform the molecular characterization of vancomycin resistant enterococci (VRE) within the faecal flora of Iberian wolf and Iberian lynx. The association with other resistance genes and the detection of virulence genes were also analysed. From 2008 to 2010, 365 faecal samples from Iberian wolf and Iberian lynx were collected and tested for VRE recovery. Mechanisms of resistance to vancomycin and other antibiotics, as well as genes encoding virulence factors were detected through PCR. Multilocus Sequence Typing (MLST) was performed for Enterococcus faecium strains. VRE were recovered in 8 of the 365 analysed samples. The vanA gene was identified in two E. faecium isolates recovered from Iberian wolf faecal samples and the remaining six showed intrinsic resistance (3 vanC1-E. gallinarum and 3 vanC2-E. casseliflavus, from Iberian wolf and Iberian lynx faecal samples, respectively). One vanA-containing isolate showed tetracycline and erythromycin resistance [with erm(B) and tet(L) genes] and the other one also exhibited ampicillin and kanamycin resistance [with erm(B), tet(M) and aph(3')-III genes]. One of the vanA-isolates revealed a new sequence type named ST573 and the other one belonged to the CC17 clonal complex (ST18). The hyl gene was detected in one E. casseliflavus and three E. gallinarum but not among vanA-positive isolates, and the occurrence of cylA and cylL genes was confirmed in two E. casseliflavus isolates. A low prevalence of VRE has been detected in faecal samples of Iberian wolf and Iberian lynx and strains with an acquired mechanism of resistance to vancomycin have not been detected among Iberian lynx.
本研究旨在对伊比利亚狼和伊比利亚猞猁粪便菌群中的万古霉素耐药肠球菌（VRE）进行分子特征分析。还分析了与其他耐药基因的关联以及毒力基因的检测。2008 年至 2010 年，收集了 365 份来自伊比利亚狼和伊比利亚猞猁的粪便样本进行 VRE 回收检测。通过 PCR 检测了对万古霉素和其他抗生素的耐药机制以及编码毒力因子的基因。对肠球菌 faecium 菌株进行了多位点序列分型（MLST）。在 365 个分析样本中，有 8 个样本回收了 VRE。从伊比利亚狼粪便样本中回收的 2 株肠球菌 faecium 中鉴定出 vanA 基因，其余 6 株表现为固有耐药（3 株为 vanC1-肠球菌 gallinarum，3 株为 vanC2-肠球菌 casseliflavus，分别来自伊比利亚狼和伊比利亚猞猁的粪便样本）。一株含 vanA 的分离株显示出四环素和红霉素耐药性（带有 erm(B)和 tet(L)基因），另一株也表现出氨苄西林和卡那霉素耐药性（带有 erm(B)、tet(M)和 aph(3')-III 基因）。一株 vanA 分离株显示出一种新的序列型，命名为 ST573，另一株属于 CC17 克隆复合体（ST18）。在一株肠球菌 casseliflavus 和三株肠球菌 gallinarum 中检测到 hyl 基因，但在 vanA 阳性分离株中未检测到，在两株肠球菌 casseliflavus 分离株中证实了 cylA 和 cylL 基因的存在。在伊比利亚狼和伊比利亚猞猁的粪便样本中检测到 VRE 的低流行率，并且在伊比利亚猞猁中未检测到对万古霉素获得性耐药的菌株。
