Oporto B, Esteban J I, Aduriz G, Juste R A, Hurtado A
Department of Animal Health and Production, NEIKER - Instituto Vasco de Investigación y Desarrollo Agrario, Berreaga 1, 48160 Derio, Bizkaia, Spain.
Zoonoses Public Health. 2008;55(2):73-81. doi: 10.1111/j.1863-2378.2007.01080.x.
Three-hundred and forty-five herds (17 swine, 122 dairy sheep, 124 beef and 82 dairy cattle) were investigated for prevalence of Shiga toxin-producing Escherichia coli (STEC). Rectal faecal samples were selectively enriched and then examined by immunodetection techniques (Immunomagnetic Separation with anti-E. coli O157 Dynabeads, ImmunoMagnetic cell Separation (IMS) and automated enzyme-linked fluorescent immunoassay using VIDAS) and polymerase chain reaction (PCR) (rfbE and fliC genes) to assess the prevalence of E. coli O157:H7. Prevalence of non-O157 STEC was estimated by PCR screening for stx genes of 10 lactose-positive colonies grown on MacConkey agar after enrichment. PCR was used on all STEC isolates to detect stx(1), stx(2), eaeA and E-hlyA genes. Both immunodetection methods showed a moderate-good level of agreement (kappa = 0.649) but IMS showed 87.5% complementary sensitivity. Prevalence of positive herds for E. coli O157:H7 was estimated at 8.7% for sheep and 3.8% for cattle, whereas all the porcine herds tested negative. Non-O157 STEC were also absent from swine, but were isolated more frequently from ovine (50.8%) than bovine herds (35.9%). Within-herd prevalences of excretion of E. coli O157:H7 established by individual testing of 279 sheep (six herds) and 30 beef cattle (one herd) were 7.3% and 6.7% respectively. PCR analysis of 49 E. coli O157:H7 and 209 non-O157 isolates showed a different distribution of virulence genes. All E. coli O157:H7 were stx(2) gene-positive, eaeA was detected in 95.9%, and the toxigenic profile stx(2)/eaeA/E-hlyA was present in 75.5% of the isolates. Among the non-O157 STEC, prevalence of eaeA was significantly lower (5.3%) and E-hlyA was present in 50.2% of the isolates but only sporadically associated with eaeA. stx(2) was predominant in non-O157 isolates from cattle, whereas in sheep the combination stx(1)/stx(2) was more prevalent. This study demonstrated the wide distribution of STEC in ruminant herds, which represent an important reservoir for strains that pose a potential risk for human infections.
对345个畜群(17个猪群、122个奶羊群、124个肉牛群和82个奶牛群)进行了产志贺毒素大肠杆菌(STEC)流行情况调查。采集直肠粪便样本进行选择性富集,然后通过免疫检测技术(使用抗大肠杆菌O157磁珠的免疫磁珠分离法、免疫磁细胞分离法(IMS)以及使用VIDAS的自动化酶联荧光免疫分析)和聚合酶链反应(PCR)(rfbE和fliC基因)来评估大肠杆菌O157:H7的流行情况。通过对富集后在麦康凯琼脂上生长的10个乳糖阳性菌落进行stx基因的PCR筛查,来估计非O157 STEC的流行情况。对所有STEC分离株进行PCR检测,以检测stx(1)、stx(2)、eaeA和E-hlyA基因。两种免疫检测方法显示出中等良好程度的一致性(kappa = 0.649),但IMS显示出87.5%的互补敏感性。估计奶羊群中大肠杆菌O157:H7阳性畜群的流行率为8.7%,奶牛群为3.8%,而所有测试的猪群均为阴性。猪群中也不存在非O157 STEC,但在羊群(50.8%)中比牛群(35.9%)中更频繁地分离到。通过对279只绵羊(6个畜群)和30头肉牛(1个畜群)进行个体检测确定的大肠杆菌O157:H7群体内排泄流行率分别为7.3%和6.7%。对49株大肠杆菌O157:H7和209株非O157分离株的PCR分析显示毒力基因的分布不同。所有大肠杆菌O157:H7均为stx(2)基因阳性,95.9%检测到eaeA,75.5%的分离株具有stx(2)/eaeA/E-hlyA产毒谱。在非O157 STEC中,eaeA的流行率显著较低(5.3%),50.2%的分离株存在E-hlyA,但仅偶尔与eaeA相关。stx(2)在牛的非O157分离株中占主导地位,而在绵羊中stx(1)/stx(2)组合更为普遍。本研究表明STEC在反刍动物畜群中广泛分布,反刍动物畜群是对人类感染构成潜在风险的菌株的重要储存宿主。