Department of Microbiology, Faculty of Veterinary Medicine, Cairo University, Cairo, Egypt.
Vector Borne Zoonotic Dis. 2012 Apr;12(4):297-305. doi: 10.1089/vbz.2010.0257. Epub 2012 Jan 9.
Some foodborne pathogens can cause mastitis, in which the organism is directly excreted into milk. Therefore, we undertook the steps to determine the prevalence and molecular characteristics of Shiga toxin-producing Escherichia coli (STEC) isolates from bovine mastitic milk in Egypt. Forty milk samples from dairy cattle showing mastitis were collected and examined for the presence of E. coli. Following enrichment and plating on selective agar, confirmation of the isolates was based on biochemical tests and the isolates were determined at the species level using cytochrome oxidase, triple sugar iron agar, urea, and indole tests as putatively E. coli. About 77.4% of the isolates belonged to four different O serogroups (O26, O86, O111, and O127). The multiplex polymerase chain reaction (PCR) found that the seven isolates revealed positive amplification of the Eagg gene from the extracted DNA of the E. coli isolates in an incidence of 100%. Also, the selected isolates were subjected to a simple PCR for the detection of 12 of the most important E. coli genes associated with virulence. Those genes detected were stx1, stx2, hylA, Flic(h7), stb, F41, K99, sta, F17, LT-I, LT-II, and eaeA. A total of seven E. coli isolates that were non-O157 isolates were investigated. Among the seven isolates, none was stx positive, and all seven lacked F41, K99, LT-I, LT-II, and Flic(h7). Of these seven isolates, three (42.85%) were enterohemorrhagic E. coli hlyA positive and two (28.57%) were eaeA positive. STEC isolates were not found in bovine mastitic milk in Egypt. Isolates from mastitic milk were potentially pathogenic for human in that they belonged to serogroups associated with diarrhea and hemolytic-uremic syndrome, and some of them were hylA, stb, sta, F17, and eaeA positive.
一些食源性致病菌可引起乳腺炎,这些病原体直接排泄到牛奶中。因此,我们着手确定埃及牛乳腺炎牛奶中产志贺毒素大肠杆菌(STEC)分离株的流行情况和分子特征。从患有乳腺炎的奶牛采集了 40 份牛奶样本进行检测,以确定是否存在大肠杆菌。在富集和选择性琼脂平板培养后,基于生化试验对分离株进行了确认,并通过细胞色素氧化酶、三糖铁琼脂、尿素和吲哚试验确定了分离株的种属水平,这些试验结果提示分离株为大肠杆菌。大约 77.4%的分离株属于四个不同的 O 血清群(O26、O86、O111 和 O127)。多重聚合酶链反应(PCR)发现,从大肠杆菌分离株提取的 DNA 中,有 7 个分离株的 Eagg 基因呈阳性扩增,阳性率为 100%。此外,还对所选分离株进行了简单 PCR 检测,以检测与毒力相关的 12 个最重要的大肠杆菌基因。检测到的基因包括 stx1、stx2、hylA、Flic(h7)、stb、F41、K99、sta、F17、LT-I、LT-II 和 eaeA。总共检测了 7 个非 O157 大肠杆菌分离株。在这 7 个分离株中,没有 stx 阳性,也没有 7 个分离株都缺乏 F41、K99、LT-I、LT-II 和 Flic(h7)。在这 7 个分离株中,有 3 个(42.85%)hlyA 阳性,2 个(28.57%)eaeA 阳性。在埃及牛乳腺炎牛奶中未发现 STEC 分离株。乳腺炎牛奶中的分离株对人类具有潜在致病性,因为它们属于与腹泻和溶血性尿毒综合征相关的血清群,其中一些分离株 hylA、stb、sta、F17 和 eaeA 阳性。