多重PCR方法检测鸭源产志贺氏毒素大肠杆菌

为了检测产志贺毒素大肠杆菌(STEC)在鸭源大肠杆菌中的分布情况,本研究建立检测STEC的多重PCR方法,针对STEC特有的毒力基因stx1、stx2、h&A和eaeA筛选了4对引物,通过对PCR反应条件的优化、特异性和灵敏度的检测,建立检测STEC的多重PCR,并应用该方法调查254株鸭源致病性E.coli和115株外表健康鸭的泄殖腔分离的E.coli中STEC的分布情况.在254株鸭源致病性E.coli中检测出6株STEC,从外表健康鸭的泄殖腔分离的115株E.coil中未检测到STEC.检出的STEC的血清型分别为O36、O60、O77、O78、O158和O7 & O92.本实验建立的检测STEC的多重PCR方法特异性好、灵敏度高;对鸭源E.coli的检测结果证实鸭致病性大肠杆菌中存在STEC菌株,分布频率较低,但其血清型具有广泛的宿主源,存在引起人类疾病的可能性.     

Shiga toxin-producing Escherichia coli O157:H7 from extensive cattle of the fighting bulls breed

Shiga toxin-producing Escherichia coli (STEC) O157:H7 represents a major public health concern worldwide, with cattle recognized as their main natural reservoir. The aim of this work was to determine the prevalence and the pheno-genotypic characteristics of STEC O157:H7 in a herd with 268 cattle of the fighting bulls breed (De Lidia breed) managed under extensive conditions in the South-West of Spain. Rectal-anal swabs of all animals were collected and examined for STEC O157:H7 by performing an immunomagnetic concentration and separation procedure combined with PCR, and the resulting isolates were characterized by both phenotypic and genotypic methods. Overall, STEC O157:H7 was isolated from seven animals (2.6%) in the herd. The PCR procedure indicated that all seven isolates displayed stx₂, eae-γ1, ehxA, O157 rfbE, and fliCh7 genes. They belonged to phage types 4 (one isolate) and 42 (two isolates), and four isolates reacted with typing phages but did not conform to a recognized pattern. Among the seven isolates there were five indistinguishable PFGE patterns and other two which differed only in ?2 restriction fragments, supporting the existence of horizontal transmission among animals in the herd. The present study demonstrates that cattle managed under extensive conditions in Spain can excrete STEC O157:H7 with their faeces. To our knowledge this is the first isolation of this pathogen from De Lidia cattle.     

新疆库尔勒地区羊源产志贺毒素大肠杆菌及其耐药性分析

为了解新疆库尔勒地区羊源产志贺毒素大肠杆菌（Shiga toxin-producing Escherichia coli,STEC）毒力基因的分布情况及其耐药性,对新疆库尔勒地区羊肉生产环节（养殖场、定点屠宰场和市场）的282份样品进行STEC的分离鉴定,并检测毒力基因的编码情况,进一步检测了17种常用抗生素的耐药情况。结果表明：羊肉生产环节样品中检出72株STEC,检出率为25.5%。72株STEC中编码毒力基因stx1+stx2、stx1、stx2、rfbE、eae、hly的数量分别为22、45、2、0、4、51个。其中,羊养殖场中存在较多的非O157 STEC,检出率为37.3%;stx以stx1为主,检出率达到了652%。72株STEC中以β-内酰胺类、氨基糖苷类、大环内酯类抗生素耐药为主,STEC菌株的多重耐药比较严重,最多可耐9种抗生素,多重耐药情况主要是耐1种和耐2种抗生素。研究表明,羊肉生产环节中羊养殖场存在大量的非O157 STEC菌株,且耐药情况比较严重,务必预防源头污染。     

Slaughterhouse effluent discharges into rivers not responsible for environmental occurrence of enteroaggregative Escherichia coli

Enteroaggregative Shiga-toxin-producing Escherichia coli strains were responsible for a massive outbreak in Europe in 2011, and had been previously isolated from French patients. The objective of this study was to investigate the presence of enteroaggregative E. coli (EAEC) in slaughterhouse effluents (wastewater, slurry, sludge and effluents), and in river waters near these slaughterhouses. A total of 10,618 E. coli isolates were screened by PCR for the presence of EAEC-associated genetic markers (aggR, aap and aatA). None of these markers was detected in E. coli isolated from slaughterhouse samples. A unique enteroaggregative E. coli (EAEC) O126:H8 was detected in river water sampled upstream from slaughterhouse effluent discharge. These results confirmed that animals might not be reservoirs of EAEC, and that further studies are required to evaluate the role of the environment in the transmission of EAEC to humans.     

Dynamics of shiga-toxin producing Escherichia coli (STEC) and their virulence factors in cattle

Starting at birth, twenty Holstein calves were housed individually, in groups of five and finally in one large freestall while fecal samples were collected weekly for 25 weeks. From each sample, twenty isolates of Escherichia coli were screened for 6 virulence markers including shiga-toxin 1, 2, intimin, enterohemolysin, the fimbrial antigen efa1 and the adhesin saa. Dynamic models of transmission of E. coli were used to model the transmission of different virulotypes between calves and the loss of the same virulotypes from the calves. It was found that, once E. coli encoding shiga-toxins in combination with enterohemolysin were transmitted and established in a calf, they tended to be eliminated less efficiently compared to E. coli without this combination of virulence markers. It was concluded that the presence of certain combinations of virulence markers coincided with persistence of E. coli in the bovine gastrointestinal tract. In addition, the combinations of stx with either eae or ehxA in E. coli have a greater impact on the loss rates than on the transmission rates.