Prevalence and level of Escherichia coli O157 on beef trimmings, carcasses and boned head meat at a beef slaughter plant

This study investigated the prevalence and level of Escherichia coli O157 on samples of beef trimmings (n=1351), beef carcasses (n=132) and bovine head meat (n=132) in a beef slaughter plant in Ireland. The survey also included an assessment of the prevalence of virulence genes in the E. coli O157 isolates obtained. Samples were examined for the presence of E. coli O157 by direct plating on SMAC-CT and by enrichment/immunomagnetic separation (IMS) with plating of recovered immunobeads onto SMAC-CT agar. Presumptive E. coli O157 isolates were confirmed by PCR targeting a range of genes i.e. vt1, vt2, eaeA, hlyA, fliC(h7) and portions of the rfb (O-antigen encoding) region of E. coli O157. Enterobacteriaceae on head meat samples were estimated by direct plating onto Violet Red Bile Glucose agar. E. coli O157 was recovered from 2.4% (32/1351) of beef trimmings samples, at concentrations ranging from<0.70-1.61 log10 cfu g(-1). Of the 32 positive isolates, 31 contained the eaeA and hylA genes while 30/32 contained the fliC(h7) gene and 31/32 contained vt1 or vt2, or both vt genes. E. coli O157 was recovered from 3.0% (4/132) of carcass samples, at concentrations ranging from <0.70-1.41 log10 cfu g(-1). All of the carcass isolates contained the eaeA, hylA and fliC(h7) genes. E. coli O157 was recovered from 3.0% (3/100) of head meat samples, at concentrations of 0.7-1.0 log10 cfu g(-1). All of the head meat isolates contained the eaeA, hylA, fliC(h7) and vt2 genes. No head meat isolates contained the vt1 gene. Head meat samples (n=100) contained Enterobacteriaceae, at concentrations ranging from 0.70-3.0 log10 cfu g(-1). Overall, the qualitative and quantitative data obtained for E. coli O157 on beef trimming samples in this study could be employed as part of a quantitative risk assessment model.     

Characterization of O157:H7 and other Escherichia coli isolates recovered from cattle hides, feces, and carcasses

In a previous study, the seasonal prevalence was reported for stx+ Escherichia coli O157:H7 in feces and on hides and carcasses of cattle at processing. Overall, 1,697 O157:H7 isolates have now been characterized for the incidence of (i) eae(O157), hlyA, stx1, and stx2 in the recovered isolates and (ii) presumptive rough and presumptive nonmotile isolates. Seven O157:H7 isolates (0.4%) lacked stx genes, although they carried eae and hlyA. All but one of the isolates carried both eae and hlyA. Approximately two-thirds of the isolates (64% when one isolate per sample was considered) carried both stx1 and stx2. E. coli O157:H7 cells that harbored both stx1 and stx2 were more often recovered from hides in the fall (79% of the fall hide isolates) and winter (84% of the winter hide isolates) than in the spring (53%) and summer (59%). Isolates recovered from preevisceration carcasses showed a similar but not statistically significant trend. Twenty-three of the 25 O157:H7 isolates carrying stx1 but not stx2 were recovered during summer. Fifteen presumptive rough and 117 presumptive nonmotile stx+ O157:H7 isolates were recovered. Ten (67%) of the presumptive rough isolates were recovered during summer. Ninety-five of the presumptive nonmotile isolates (81%) were recovered during fall. Forty-eight percent of the false-positive isolates (175 of 363) tentatively identified as O157:H7 were O157+ H7- and lacked eae(O157), hlyA, and stx. These data suggest that in beef processing samples (i) there are minor seasonal variations in the prevalence of stx genes among E. coli O157:H7 isolates, (ii) presumptive rough and presumptive nonmotile stx+ O157:H7 isolates are present, (iii) E. coli O157:H7 isolates lacking stx genes may be rare, and (iv) O157+ H7- isolates lacking stx genes can result in many false-positive results.     

Isolation and characterization of Shiga toxin-producing Escherichia coli O157:H7 and non-O157 from beef carcasses at a slaughter plant in Mexico

The contamination of beef carcasses with Shiga toxin-producing O157:H7 and non-O157 Escherichia coli (STEC) obtained from a slaughter plant in Guadalajara, Mexico was investigated. A total of 258 beef carcasses were sampled during a 12-month period. All samples were assayed for STEC by selective enrichment in modified tryptone soy broth supplemented with cefixime, cefsulodin and vancomycin, followed by plating on Sorbitol MacConkey Agar supplemented with cefixime and tellurite (CT-SMAC). Simultaneously, all samples were assayed by immunomagnetic separation (IMS) and plated on CT-SMAC and CHROMagar. The presence of the stx1, stx2, eaeA and hly933 genes, recognized as major virulence factors of STEC, was tested for O157:H7 and non-O157 E. coli isolates by multiplex polymerase chain reaction (PCR). STEC was detected in two (0.8%) samples. One of these STEC isolates corresponded to the serotype O157:H7 showing stx2, eaeA and hyl933 genes. The other isolate corresponded to non-O157 STEC and only had the stx1 gene. Thirteen carcasses (5%) were positive for nonmotile E. coli O157 and 7 (2.7%) were positive for E. coli O157:H7. The presence of O157:H7 and non-O157 STEC on beef carcasses in this slaughter plant in Guadalajara, Mexico, emphasizes the importance of implementing the Hazard Analysis and Critical Control Point (HACCP) system, as well as the need for implementing, evaluating, and validating antimicrobial interventions to reduce the presence of potential pathogenic microorganisms.     

A multiplex polymerase chain reaction assay for rapid detection and identification of Escherichia coli O157:H7 in foods and bovine feces

A multiplex polymerase chain reaction (PCR) assay was designed to simplify detection of Escherichia coli O157:H7 and to identify the H serogroup and the type of Shiga toxin produced by this bacterium. Primers for a plasmid-encoded hemolysin gene (hly933), and chromosomal flagella (fliCh7; flagellar structural gene of H7 serogroup), Shiga toxins (stx1, stx2), and attaching and effacing (eaeA) genes were used in a multiplex PCR for coamplification of the corresponding DNA sequences from enterohemorrhagic E. coli (EHEC) O157:H7. Enrichment cultures of ground beef, blue cheese, mussels, alfalfa sprouts, and bovine feces, artificially inoculated with various levels of E. coli O157:H7 strain 933, were subjected to a simple DNA extraction step prior to the PCR, and the resulting amplification products were analyzed by agarose gel electrophoresis. Sensitivity of the assay was < or = 1 CFU/g of food or bovine feces (initial inoculum level), and results could be obtained within 24 h. Similar detection levels were obtained with ground beef samples that underwent enrichment culturing immediately after inoculation and samples that were frozen or refrigerated prior to enrichment. The multiplex PCR facilitates detection of E. coli O157:H7 and can reduce the time required for confirmation of isolates by up to 3 to 4 days.     

Detection, occurrence, and characterization of Escherichia coli O157:H7 from raw ewe's milk in Spain

A study was carried out in the Castilla y León region of Spain to investigate the presence of Escherichia coli O157:H7 in raw ewe's milk samples collected from several cheese factories during 1 year. All specimens were examined for E. coli O157:H7 by selective enrichment at 41.5 +/- 1.0 degrees C, after both 6 and 22 h of incubation, and then immunomagnetically separated and plated on cefixime-potassium tellurite-sorbitol MacConkey agar. No growth was obtained in the enrichment broth after a 6-h incubation. Presumptive colonies obtained after 22 h of incubation were screened by a multiplex PCR assay for the presence of rfbO157 and fliCH7 genes. Of all the ewe's milk samples studied, three were positive for E. coli O157:H7. The E. coli O157:H7 strains that were positive for the rfbO157 and fliCH7 genes were then analyzed by multiplex PCR for the presence of virulence genes (stx1, stx2, ehxA, and eaeA). All E. coli O157:H7 isolates were Shiga toxigenic and harbored additional genes related to virulence (ehxA and eaeA). The predominant Stx toxin type was stx2. These results demonstrate that raw ewe's milk used in cheesemaking may be sporadically contaminated with E. coli O157:H7 strains that are potentially pathogenic for humans.     

Isolation and characterization of verocytotoxin-producing Escherichia coli O157 from Turkish cattle

The objective of this study was to collect rectal swabs from the cattle in a slaughterhouse located in Hatay (Turkey) immediately after slaughter for the isolation and characterization of verotoxin-producing Escherichia coli O157 in each month during a 1-year period. The rectal swab samples were analyzed for the isolation of E. coli O157 through pre-enrichment, immunomagnetic separation and selective plating on CT-SMAC agar. E. coli O157 was isolated from 77 (13.6%) of the samples. The presence of E. coli O157 changed during a 1-year period, in that the occurrence of E. coli O157 was the highest in July and November and lowest in February. A total of 66 isolates out of 77 were serotype O157:H7 and 11 were serotype O157:NM. PCR analysis of E. coli O157 virulence genes revealed that all O157:H7/NM were positive for rbf(O157), 74 positive for EhlyA, 72 positive for eaeA, 62 positive for vtx2, and 3 positive for both vtx1 and vtx2. It was presented by cytotoxicity tests that many of E. coli O157 isolates showed high cytotoxicity on Vero cells. All of the isolates containing EhlyA showed enterohaemolysin production.     

Detection and frequency of VT1, VT2 and eaeA genes in Escherichia coli O157 and O157:H7 strains isolated from cattle, cattle carcasses and abattoir environment in Istanbul

The aim of this study was to detect VT1, VT2 and eaeA genes and to determine the frequency of these genes in Escherichia coli O157 and O157:H7 strains isolated from cattle, cattle carcasses and environmental samples of the 5 abattoirs located in Istanbul, Turkey. For this, the presence of VT1, VT2 and eaeA genes in 26 strains of E. coli O157:H7 and 6 strains of O157 was investigated by multiplex-PCR. The results have shown that eaeA gene was detected in all O157 and O157:H7 strains tested. Both VT2 and eaeA genes were detected in 4 (80%) of 5 strains of E. coli O157 and eaeA alone in 1 strain of O157. In 27 strains of O157:H7, 5 (18.5%) strains were found to be positive for VT1, VT2 and eaeA genes, 19 (70.3%) strains for both VT2 and eaeA and, 3 (11.1%) strains for only eaeA gene. Either VT1 alone or VT2 alone was not detected in any strains tested. eaeA gene alone in 2 strains, VT2-eaeA genes in 9 strains and VT1-VT2-eaeA genes in 2 strains were detected in 13 of E. coli O157:H7 strains isolated from cattle. eaeA alone in 1 strain, VT2-eaeA genes in 5 strains and VT1-VT2-eaeA genes in 2 strains were detected in 8 of E. coli O157:H7 strains isolated from carcasses. VT2-eaeA genes in 5 strains (isolated from hands, apron, knife and floor) and VT1-VT2-eaeA genes in 1 strain (isolated from knife) were also detected in 6 of E. coli O157:H7 strains isolated from environmental samples. This study reveals that most of the strains are found to be toxigenic and it is most likely that strains isolated from carcasses and abattoir environment originated from cattle feces. Therefore, HACCP systems are necessary from farm to table especially in the abattoirs to prevent contamination of meat and abattoir environment with intestinal content.     

Molecular characterization of Escherichia coli O157:H7 hide contamination routes: feedlot to harvest

This study was conducted to identify the origin of Escherichia coli O157:H7 contamination on steer hides at the time of harvest. Samples were collected from the feedlot, transport trailers, and packing plant holding pens and from the colons and hides of feedlot steers. A total of 50 hide samples were positive for E. coli O157:H7 in two geographical locations: the Midwest (25 positive hides) and Southwest (25 positive hides). Hide samples were screened, and the presence of E. coli O157: H7 was confirmed. E. coli O157:H7 isolates were fingerprinted by pulsed-field gel electrophoresis and subjected to multiplex PCR procedures for amplification of E. coli O157:H7 genes stx1, stx2, eaeA, fliC, rfbEO157, and hlyA. Feedlot water trough, pen floor, feed bunk, loading chute, truck trailer side wall and floor, packing plant holding pen floor and side rail, and packing plant cattle drinking water samples were positive for E. coli O157:H7. Pulsed-field gel electrophoresis banding patterns were analyzed after classifying isolates according to the marker genes present and according to packing plant. In this study, hide samples positive for E. coli O157:H7 were traced to other E. coli O157:H7-positive hide, colon, feedlot pen floor fecal, packing plant holding pen drinking water, and transport trailer side wall samples. Links were found between packing plant side rails, feedlot loading chutes, and feedlot pens and between truck trailer, different feedlots, and colons of multiple cattle. This study is the first in which genotypic matches have been made between E. coli O157:H7 isolates obtained from transport trailer side walls and those from cattle hide samples within the packing plant.     

Prevalence and characterization of Escherichia coli O157 isolates from Minnesota dairy farms and county fairs

Samples were collected from 26 organic and conventional farms and 12 county fairs in Minnesota during 2001 and 2002 to identify the presence of Escherichia coli O157. Immunomagnetic separation was used for isolation of E. coli O157. Isolates were further characterized by the presence of virulence marker genes (stx1, stx2, eaeA, E-hly, katP, etpD, and espP), antimicrobial susceptibility profiles, and genotypes. During 2001, E. coli O157 was isolated from 16 (5.2%) of 305 fecal samples and from 7 (36.8%) of 19 farms. During 2002, E. coli O157 was isolated from 6 (4.5%) of 132 fecal samples from weaned calves at 4 (23.5%) of 17 farms. During 2001 and 2002, cattle manure samples were collected from 12 county fairs, and E. coli O157 was isolated from 19 (11%) of 178 samples and 9 (75%) of 12 county fairs. Among 40 E. coli O157 isolates, 17 isolates (43%) had both the stx1 and stx2 genes, and 21 strains (53%) had the stx2 gene only. Thirteen percent of O157 isolates were resistant to tetracycline, and 25% were resistant to sulfadimethoxine. Heterogeneity of E. coli O157 strains was demonstrated by the presence of 22 different pulsed-field gel electrophoresis (PFGE) patterns. Four PFGE patterns matched those of isolates previously found in humans. The presence of E. coli O157 at county fairs suggests the potential for transmission to the public, who may have contact with cattle or their environment.     

Detection of virulence-associated genes in Escherichia coli O157 and non-O157 isolates from beef cattle, humans, and chickens

Food-producing animals can be reservoirs of pathogenic Escherichia coli strains that can induce diseases in animals or humans. Contamination of food by E. coli O157:H7 raises immediate concerns about public health, although it is not clear whether all E. coli O157 isolates of animal origin are equally harmful to humans. Inversely, the pathogenic potential of atypical E. coli O157 isolates and several non-O157 serotypes often is ignored. We used a DNA microarray capable of detecting a subset of 346 genes to compare the virulence-associated genes present in eight E. coli O157 isolates from human cases, 14 antibiotic-resistant and/or hypermutable E. coli O157 isolates from beef cattle, and four antibiotic-resistant, sorbitol-negative, non-O157 E. coli isolates from healthy broiler chickens. Hybridization on arrays (HOA) revealed that O157 isolates from beef cattle and humans were genetically distinct, although they possessed most of the same subset of virulence genes. HOA allowed discrimination between hypermutable and antibiotic-resistant O157 isolates from beef cattle based on hybridization results for the stx2 and ycgG genes (hypermutable) or ymfL, stx1, stx2, and hlyE(avian) genes (resistant). However, the absence of hybridization to gene yfdR characterized human isolates. HOA also revealed that an atypical sorbitol-fermenting bovine O157 isolate lacked some genes of the type 3 secretion system, plasmid pO157, and the stx1 and stx2 genes. This isolate had a particular pathotype (eaeA(beta) tir(alpha) espA(alpha) espB(alpha) espD(alpha)) not found in typical E. coli O157:H7. HOA revealed that some non-O157 E. coli isolates from healthy chickens carried genes responsible for salmochelin- and yersiniabactin-mediated iron uptake generally associated with pathogenic strains.     

Testing of swine feces obtained through the National Animal Health Monitoring System's Swine 2000 study for the presence of Escherichia coli O157:H7

Fecal samples collected from healthy pigs from 13 of the top 17 swine-producing states were tested for Escherichia coli O157:H7 as part of the National Animal Health Monitoring System Swine 2000 study. Serogroup O157 strains were isolated from 106 of 2,526 fecal samples. None of the isolates were positive by PCR for the fliCh7 (H7 flagellin) gene or for the hly933 (hemolysin) gene; however, one isolate was positive for the stxl gene (Shiga toxin 1), an additional four isolates were positive for the stx2 gene (Shiga toxin 2), and three isolates possessed the eae gene (intimin).     

Escherichia coli O157 in cattle and sheep at slaughter, on beef and lamb carcasses and in raw beef and lamb products in South Yorkshire, UK

A 1 year study of Escherichia coli O157 in cattle and sheep at slaughter, on beef and lamb carcasses and in raw beef and lamb products from retail butchers' shops was performed in the Sheffield area. Each month, samples of rectal faeces were collected immediately after slaughter from 400 cattle and 600 sheep, and 400-430 samples of raw meat products were purchased from butchers' shops. Meat samples were also obtained from 1500 beef and 1500 lamb carcasses. All samples were examined for E. coli O157 by enrichment culture, immunomagnetic separation and culture of magnetic particles onto cefixime tellurite sorbitol MacConkey agar. Raw meat products were also examined for numbers of generic E. coli by a standard membrane culture method. E. coli O157 was isolated from 620 (12.9%) of 4800 cattle, 100 (7.4%) of 7200 sheep, 21 (1.4%) of 1500 beef carcasses, 10 (0.7%) of 1500 lamb carcasses and from 22 (0.44%) of 4983 raw meat products. E. coli O157 was isolated more frequently from lamb products (0.8%) than from beef products (0.4%). Numbers of generic E. coli in meat products reached seasonal peaks in July and August with counts of > 10(4)/g occurring more frequently in lamb products (50.8 and 42.4%, respectively) than in beef products (19.3 and 23.8%, respectively). The majority of E. coli O157 strains, from animals, carcasses and meat samples, were isolated during the summer. Most were verocytotoxigenic as determined by Vero cell assay and DNA hybridisation, eaeA gene positive and contained a 92 kb plasmid. The isolates were compared with 66 isolates from human cases over the same period. A combination of phage type, toxin genotype and plasmid analysis allowed subdivision of all the E. coli O157 isolates into 96 subtypes. Of these subtypes, 53 (55%) were isolated only from bovine faecal samples. However, 61 (92%) of the 66 isolates from humans belonged to 13 subtypes which were also found in the animal population.     

MPCR检测食品中大肠杆菌O157和单核增生李斯特氏菌的研究

根据大肠杆菌O157(Escherichia coli O157,E. coli O157)的志贺样毒素基因slt和“黏附抹平”因子eaeA基因、单核增生李斯特氏菌(Listeria monocytohenes,LM)的编码溶血素O的hlyA基因和毒力基因plcA,分别设计上游和下游的slt、eaeA、hlyA和plcA四对引物,对应扩增片段依次为780、450、708、600bp。通过对单管多重PCR(multiplex PCR,MPCR)扩增的特异性、敏感性分析以及对单管多重PCR扩增条件,如Mg2+浓度、dNTPs浓度和退火温度等的优化,建立快速检测大肠杆菌O157和单增李斯特菌的多重PCR方法,该方法能同时检测到0.50ng的E. coliO157和单增李斯特菌基因组DNA,并且操作简便、快速,具有良好的敏感性和特异性,能够快速实现对食品中多种致病菌的诊断和监控。     

Seasonal prevalence of Escherichia coli O157:H7 in beef cattle feces

Cattle are an asymptomatic reservoir of Escherichia coli O157:H7, but the bacterial colonization and shedding patterns are poorly understood. The prevalence and shedding of this human pathogen have been reported to be seasonal with rates typically increasing during warm months. The objectives of this study were (i) to assess the prevalence of E. coli O157:H7 in feces of feedlot cattle in Kansas during summer, fall, and winter months, and (ii) to characterize E. coli O157:H7 by screening for virulence factors. Of 891 fecal samples collected, 82 (9.2%) were positive for E. coli O157:H7. No significant differences in prevalence were detected among summer, fall, and winter months. The highest monthly prevalence (18.1%) was detected in February. All tested isolates were positive for stx2 (Shiga toxin 2) and eaeA (intimin) genes; 14 isolates (12.8%) also carried stx1. Our results indicate the prevalence of E. coli O157:H7 in beef cattle feces is not necessarily season dependent.     

Evaluation of molecular typing methods for Escherichia coli O157:H7 isolates from cattle, food, and humans

Escherichia coli O157:H7, a Shiga toxin-producing E. coli, has been the causative agent of many cases of severe, often life-threatening foodborne illness. Because of the importance of E. coli O157:H7 to public health, many molecular typing methods have been developed to determine its transmission routes and source of infection during epidemiological investigations. Pulsed-field gel electrophoresis (PFGE) is currently used by public health organizations to track infections of E. coli O157:H7 and other foodborne pathogens. In this study, we compared the ability of PFGE, multilocus sequence typing (MLST), and repetitive-element PCR (Rep-PCR) to distinguish among 92 E. coli O157:H7 isolates from cattle, food, and infected humans. Several virulence genes, including the intimin gene (eaeA), the hemolysin gene (hlyA), and the H7 fimbrial gene (fliC), and a housekeeping gene for beta-glucuronidase (uidA) were included in MLST. Rep-PCR reactions were performed using a commercially available typing kit (Bacterial Barcodes Inc., Houston, Tex.) with the provided Uprime-RI primer set. Results of the study indicated that PFGE provided the most discrimination among the techniques, identifying 72 distinct PFGE profiles for the isolates; Rep-PCR elucidated 14 different profiles, whereas MLST generated five profiles. Additionally, there did not appear to be any correlation among the typing methods examined in this study. Therefore, to date, PFGE remains the technique of choice for molecular subtyping of E. coli O157:H7.     

2005-2007年我国食品中大肠埃希菌O157分离株tccP1基因的分布研究

目的 了解我国2005-2007年食源性疾病监测网分离的大肠埃希菌O157分离株中tccP1基因的分布情况及特点.方法 对89株食源性大肠埃希菌O157分离株运用PCR方法进行检测.结果 tccP1基因的阳性率为55.1%.结论 tccP1基因广泛存在于O157:H7大肠埃希菌中,在O157:非H7中检出率很低.其分布...     

Prevalence of Escherichia coli O157:H7 in the American bison (Bison bison)

Bison is becoming a popular meat source for consumers, but very little is known about the bison's status with respect to Escherichia coli O157:H7. We conducted a study to determine the prevalence and identify virulence genes and pulsed-field genetic types of E. coli O157:H7 in bison. Rectal contents and rectoanal mucosal swab (RAMS) samples were collected from a total of 342 bison at slaughter on seven different dates. Isolation of E. coli O157:H7 was by enrichment, immunomagnetic separation, and plating on selective medium, and identification was based on sorbitol fermentation reaction, indole production, and O157 agglutination test. An overall E. coli O157:H7 prevalence of 47.4% was observed. Fecal prevalence across sampling days ranged from 17 to 83%, with an average of 42.1%. The prevalence in the rectoanal mucosal region ranged from 2.2 to 50%, with an average of 19.9%. All E. coli O157:H7 isolates (n = 212) possessed eae, hlyA, fliC, and stx2 genes. The antiterminator Q gene, Q933, was present in 50.7% of fecal and 38% of RAMS isolates, and Q21 was present in 52.1% of fecal and 61.5% of RAMS isolates. The pulsed-field gel electrophoresis analysis of isolates revealed 11 types (> 95% Dice similarity) and 19 subtypes (100% Dice similarity). Two pulsed-field genetic types accounted for 76.4% of total isolates. Our study suggests that the prevalence of E. coli O157:H7 in rectal contents or on rectal mucosa of bison is variable, but relatively high overall and bison could serve as an important reservoir for human infection.     

Antimicrobial resistance in Irish isolates of verocytotoxigenic Escherichia coli (E. coli)--VTEC

This study compared the antimicrobial resistance profiles of Escherichia coli O157:H7 isolates (n=257) recovered from bovine hides, minced beef and human clinical samples in Ireland, to those profiles of a range of Irish non-O157 E. coli (O111 and O26) isolates (n=31) from a variety of clinical and veterinary sources. Four multi-drug resistant (MDR) E. coli O157:H7 food isolates were identified, with resistance to 10 (1 isolate), 6 (1 isolate) and 4 (2 isolates) antimicrobial agents, respectively. Two of these isolates (resistant to 7 and 4 antimicrobial classes) were characterised further by molecular methods and found to contain class 1 integrons along with a beta-lactamase-encoding tem-1 gene. Transfer of antimicrobial resistance (ampicillin, streptomycin and sulphonamides), the tem-1 gene and markers (int1, qacEDelta1, sul1) characteristic of class 1 integrons were evident in one MDR isolate (resistant to 4 antimicrobial classes) when conjugation and transformation experiments were performed. A clinical isolate and a veterinary isolate of the O111 serotype were MDR and resistant to 4 and 3 antimicrobial classes, respectively. These data suggest that the prevalence of antimicrobial resistance among the three VTEC serotypes examined in this study is low. However, these organisms may become a public health risk should they enter the food chain.     

肉类及蔬菜食品中EHEC O157污染分布及分型研究

为深入了解肉类及蔬菜中肠出血性大肠埃希氏菌(EHEC O157)的污染分布规律和遗传多样性,随机采集全国18个城市的食品样品,参考GB/T 4789.36-2008方法进行样品处理,并用rfbE/fliCH7双重PCR对菌株进行鉴定;分别采用单重PCR和ERIC-PCR对菌株进行毒力基因(eae、hlyA、stx1和stx2)检测和分子分型。结果表明,414份样品中检出18份EHEC O157阳性样品,总污染率4.35%,其中生鲜肉12份,速冻肉6份,蔬菜未检出;经过生化、血清鉴定和双重PCR检测,鉴定出52株EHEC O157,包括29株O157:H7,3株O157:NM(fliCH7+,无动力),2株O157:NM(fliCH7-,无动力)和18株O157:hund(未确定H型);毒力基因检测结果发现,52株菌株中有50株携带毒力基因,其中40株(76.92%)携带eae,31株(59.62%)携带hlyA,20株(38.46%)携带stx1,24株(46.15%)携带stx2。ERIC-PCR分型结果表明,在相似系数为0.80时,菌株可分为12个聚类簇,F型为其主要基因簇,分离株的基因型呈现多样性。     

温州市食品中肠出血性大肠杆菌O157∶H7污染状况调查

目的调查温州市食品中肠出血性大肠杆菌O157∶H7的污染状况,了解肠出血性大肠杆菌O157∶H7毒力因子的携带与耐药情况。方法采用分层随机抽样的方法,对2008—2011年抽取的231份食品样品,用免疫磁珠富集法对大肠杆菌O157∶H7进行分离,对该分离株进行生化、血清学鉴定,以纸片法(K-B法)进行药敏试验;采用PCR方法检测O、H抗原基因和stx1、stx2、eaeA、hlyA等4种毒力基因。结果 231份样品中,分离出1株肠出血性大肠杆菌O157∶H7,检出率为0.43%;O157抗原和H7抗原的核酸检测结果阳性,但未检测到stx1、stx2、eaeA、hlyA等4种毒力基因,菌株对红霉素、利福平、150μg/片和10μg/片两种浓度的0/129(二氨基二异丙基喋啶磷酸盐)耐药。结论温州市食品中存在大肠杆菌O157∶H7的污染,检出率较低,但提示要加强主动监测,通过有效干预,防范肠出血性大肠杆菌O157∶H7所致食源性疾病的发生。