Escherichia coli O157 and non-O157 Shiga toxin-producing Escherichia coli in fecal samples of finished pigs at slaughter in Switzerland

Fecal samples from 630 slaughtered finisher pigs were examined by PCR to assess the shedding of Escherichia coli O157 (rfbE) and Shiga toxin-producing E. coli (STEC, stx). The proportion of positive samples was 7.5% for rfbE and 22% for stx. By colony hybridization, 31 E. coli O157 and 45 STEC strains were isolated, and these strains were further characterized by phenotypic and genotypic traits. Among E. coli O157 strains, 30 were sorbitol positive, 30 had an H type other than H7, and none harbored stx genes. Intimin (eae), enterohemolysin (ehxA), EAST1 (astA), and porcine A/E-associated protein (paa) were present in 10, 3, 26, and 6% of strains. Among them, one eae-gamma1-positive O157:H7 strain testing positive for ehxA and astA and two eae-alpha1-positive O157:H45 strains were classified as enteropathogenic E. coli (EPEC). The O157:H45 EPEC harbored the EAF plasmid and the bfpA gene, factors characteristic for typical EPEC. The isolated STEC strains (43 sorbitol positive) belonged to 11 O:H serotypes, including three previously reported in human STEC causing hemolytic uremic syndrome (O9:H-, O26:H-, and O103:H2). All but one strain harbored stx2e. The eae and ehxA genes, which are strongly correlated with human disease, were present in only one O103:H2 strain positive for stx1 and paa, whereas the astA gene was found more frequently (14 strains). High prevalence of STEC was found among finisher pigs, but according to the virulence factors the majority of these strains seem to be of low virulence.     

The prevalence of Escherichia coli O157 and O157:H7 in ground beef and raw meatball by immunomagnetic separation and the detection of virulence genes using multiplex PCR

The present study was conducted to investigate the presence of Escherichia coli O157 and O157:H7 strains and to detect the presence of the stx1, stx2, and eaeA genes in isolates derived from 200 samples (100 samples from fresh ground beef and 100 samples from raw meatball). The samples were purchased from the Samsun Province in Turkey, over a period of 1 year. Enrichment-based immunomagnetic separation and multiplex polymerase chain reaction were applied for these analyses. E. coli O157 was detected in five of the 200 (2.5%) samples tested (one isolated from ground beef and four from meatball samples), whereas E. coli O157: H7 was not detected in any sample. During the analysis, eight strains of E. coli O157 were obtained. The genes stx1, stx2, and eaeA were detected in two E. coli O157 isolates obtained from two meatball samples, whereas only the eaeA and the stx2 genes were detected in four E. coli O157 strains that were isolated from one meatball sample. None of the stx1, stx2, and eaeA was detected in the E. coli O157 isolates obtained from the ground beef and the one meatball samples.     

Phenotypic and genotypic characterisation of Escherichia coli strains serogrouped as enteropathogenic E. coli (EPEC) isolated from pasteurised milk

Fifty-six Escherichia coli strains, serogrouped as EPEC, isolated from three different brands of pasteurised milk commercialised in Rio de Janeiro, Brazil, were tested for enteropathogenicity markers. Most of the strains (71.4%) were adherent to HEp-2 cells. The adherent strains were distributed among 7 EPEC serogroups (O26, O55, O111, O114, O125, O127, O128, O158). Although almost half of these strains (33.9%) presented unrecognisable adherence phenotypes, classical adherence patterns (localised-like, aggregative and diffuse adherence) described for E. coli and epidemiologically associated with diarrheagenic strains were observed. None of the strains showed typical localised adherence, usually associated with EPEC strains, but 4 of them displayed a localised-like adherence (LAL) phenotype, characterised by fewer and less compact microcolonies but that is still associated with diarrheagenic strains as well as strains of non-human origin. Indeed, 3 of these 4 strains were able to elicit the attaching-effacing lesion (FAS-positive), the central feature of EPEC pathogenesis, and hybridised with bfpA and eae DNA probes. The other LAL-positive strain hybridised with the bfpA probe but gave negative results for the eae probe and FAS assays. Interestingly, all LAL-positive strains produced amplicons of 200 bp in the PCR for bfpA, instead of the expected 326 bp fragment. PCR reactions for stx1 and stx2, two shiga-toxin-encoding genes, gave negative results. Typing of LEE-associated genes by PCR showed the profile eae (beta), tir (beta), espA (alpha) and espB (alpha) for one of the LAL-positive strain. The most prevalent adherence phenotype was the aggregative pattern which is observed in strains epidemiologically associated with persistent diarrhea. Additionally, one strain promoted complete detachment of the Hep-2 cell monolayer after 3 h of infection which might be related to the production of citotoxins, a feature that has been increasingly observed in clinical strains. The possession of EPEC-related O and H antigens is no longer deemed an essential characteristic of true pathogenic EPEC strains, emphasising the importance of routinely screen for virulence markers in E. coli strains isolated from foods. Our results are in accordance with data from the literature that demonstrate that environmental strains display atypical features but yet are capable of eliciting the classical A/E lesion and thus must be considered as potentially pathogenic. Further, our results demonstrate the potential of pasteurised milk as a vehicle for transmission of diarrheagenic E. coli in Brazil.     

Fecal shedding of Escherichia coli O157, Salmonella, and Campylobacter in Swiss cattle at slaughter

Fecal samples from 2,930 slaughtered healthy cattle were examined with the following goals: (i) to monitor the shedding of Escherichia coli O157, Salmonella, and Campylobacter in cattle; and (ii) to further characterize the isolated strains. The percentage of the 2,930 samples that tested positive for E. coli O157 by PCR was 1.6%. Thirty-eight strains from different animals that agglutinated with Wellcolex E. coli O157 were isolated. Of the six sorbitol-negative strains, five tested positive for stx2 genes (two times for stx2c and three times for stx2), and one strain tested positive for stx1 and stx2c genes. All sorbitol-negative strains belonged to the serotypes O157:H7- and O157:H7 and harbored the eae type gamma 1 and ehxA genes. The 32 sorbitol-positive strains tested negative for stx genes and belonged to the serotypes O157:H2, O157:H7, O157:H8, O157:H12, O157:H19, O157:H25, O157:H27, O157:H38, O157:H43, O157:H45, and O157:H-. All O157:H45 strains harbored the eae subtype alpha 1 and therefore seem to be atypical enteropathogenic E. coli strains. Whereas none of 1,000 examined samples was positive for Salmonella, 95 of 935 (10.2%) samples were positive for Campylobacter, and all strains were identified as C. jejuni. Sixteen Campylobacter strains were resistant to tetracycline, five were resistant to nalidixic acid/ciprofloxacin, four were resistant to streptomycin, and one was resistant to nalidixic acid/ciprofloxacin and streptomycin. Fecal shedding of zoonotic pathogens in slaughter animals is strongly correlated with the hazard of carcass contamination. Therefore, the maintenance of slaughter hygiene is of crucial importance.     

Diarrheagenic Escherichia coli detected by 16-plex PCR in raw meat and beef intestines sold at local markets in Ouagadougou, Burkina Faso

The study investigated the prevalence of five major Escherichia coli pathogroups in raw meats and beef intestines sold at the local markets in Ouagadougou, Burkina Faso. One hundred and twenty samples (36 beef, 36 beef intestine, 24 mutton and 24 chicken samples) were purchased from four markets between October 2008 and February 2009. Fifteen virulence genes specific for Shiga toxin-producing E. coli (STEC), enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC) and enteroaggregative E. coli (EAEC) were examined using 16-plex PCR for mixed bacterial cultures derived from the samples. One or more diarrheagenic E. coli pathogroup was detected in 51 (43%) of all the 120 samples: in 16 (44%) beef, 19 (53%) beef intestine, 9 (38%) mutton and in 7 (29%) chicken samples. Thirty three (28%) samples were positive for stx(1) and/or stx(2) indicating presence of STEC. EPEC virulence markers (eae, escV and/or ent and/or bfp and/or EHEC-hlyA) were detected in 14 (12%) stx-negative samples. ETEC virulence markers (elt and/or estIb and/or estIa) were detected in 10 (8%) samples and EAEC virulence markers (pic or aggR) in 5 (4%) samples. No EIEC was detected. The results show that in Burkina Faso the microbiological quality of retail meat is alarmingly poor due to the common occurrence of diarrheagenic E. coli bacteria.     

Incidence of Escherichia coli O157:H7 and E. coli virulence factors in US bulk tank milk as determined by polymerase chain reaction

Samples of bulk tank milk from dairies across the United States, taken as part of the National Animal Health Monitoring Dairy 2002 survey, were analyzed for the presence of several genes encoding virulence factors associated with enterohemorrhagic forms of Escherichia coli (EHEC) using real-time and conventional PCR assays. Samples from 859 farms in 21 states were collected and enriched in EC medium at 42.5°C to amplify any E. coli present, and DNA was isolated from the resulting biomass. The eaeA gene encoding intimin, a virulence factor associated with enteropathogenic forms of E. coli and EHEC, was detected in 199 (23%) of the samples. Thirty-six samples (4.2%) were positive for eaeA, the gamma allele of the translocated intimin receptor (γ-tir), found in EHEC strains of O157:H7, and one or both shiga-like toxin genes (stx1 and stx2), a combination that may be indicative of the presence of O157:H7 EHEC. Testing these 36 samples with a commercially available real-time PCR kit for detection of O157:H7 indicated that 5 samples could be contaminated with O157:H7. A multiplex PCR to detect the presence of fliC, rfbE, and hlyA genes found in O157:H7 reduced to 2 (0.2% of all samples) the number of samples likely to be contaminated with this organism. A strain of O157:H7 (eaeA⁺, γ-tir⁺, stx2⁺, rfbE⁺, fliC⁺, hlyA⁺) was subsequently isolated from one sample. Thirty-four eaeA-positive samples did not contain detectable γ-tir but did contain one or both of the stx genes suggesting the presence of EHEC strains other than O157:H7. These results indicate a low incidence of O157:H7 in bulk tank milk but suggest that a risk from other enteropathogenic and EHEC forms of E. coli may exist and that PCR targeting virulence factors associated with highly pathogenic forms of E. coli may be an effective means of detecting potential dangers in raw milk.     

A new 7-plex PCR assay for simultaneous detection of shiga toxin-producing Escherichia coli O157 and Salmonella Enteritidis in meat products

A 7-plex PCR assay was developed to achieve an effective detection and identification of serotype Enteritidis of Salmonella spp. and shiga toxin-producing type of Escherichia coli O157 in meat products. Six DNA sequences in the invA and sdfI genes of Salmonella Enteritidis as well as rfbE, eae, stx1, and stx2 genes of E. coli O157:H7 were employed to design primers. The multiplex PCR assay could specifically and sensitively detect and identify target pathogens. Applying the assay to meat samples, the multiplex PCR assay was able to simultaneously detect and identify the two pathogens at a sensitivity of three CFU/10?g raw meats after simple 16?h enrichment in buffered peptone water. Further applying in 21 retail meat samples revealed that two samples were positive for non-shiga toxin producing E. coli O157, one sample was positive for Stx2 producing E. coli O157 and five samples were positive for Salmonella enterica Enteritidis. Taken together, the 7-plex PCR assay is a rapid and reliable method for effectively screening single or multiple pathogens occurrences in various meat products, and could also identify the Salmonella enterica Enteritidis from all Salmonella spp. and shiga toxin producing type from all E. coli strains. Considering as a non expensive screening tool, the multiplex PCR assay has a great potential in complement for food stuff analysis by conventional microbiological tests.     

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 the Shiga toxin gene (stx)-bearing Escherichia coli O157 and non-O157 from retail meats in Shandong Province, China, and characterization of the O157-derived stx2 phages

Infection by Shiga toxin (Stx)-producing Escherichia coli of non-O157 and O157 serotypes are rare in China, but infection by O157 serotype was found in Shandong Province and three other provinces in China. To understand the reason for these rare infections and to determine the safety of retail meats in Shandong Province, we examined the distribution of Shiga toxin gene (stx)-bearing E. coli in retail meats and characterized the isolated stx-bearing strains. We used hybridization with DNA probes and isolated stx1- and/or stx2-positive E. coli from 31 (58%) of 53 retail meat samples, with beef showing the highest frequency (68%). Of 42 stx-positive isolates, none belonged to O157. Using the O157-specific immunomagnetic bead technique, we isolated E. coli O157 carrying the eae and stx2 genes from eight beef samples (26%). These strains produced little or no Stx2 and carried a unique q gene. Replication of the stx2 phages was detected in these strains, whereas stx2 phage replication was not detected in our previous study in which we examined similar stx2-bearing E. coli O157 strains from other Asian countries. Analysis of E. coli C600 lysogenized with the stx2 phages found in this study suggests that the lack of Stx2 production is due to changes in non-q gene region(s) of the phage genome or chromosomal mutation(s) in the host. Our data and reports by other workers suggest it is necessary to determine if various stx2-bearing E. coli O157 strains producing Stx2 to varying degrees are distributed in meats in various locations in China.     

Extended-spectrum β-lactamase,shigatoxin and haemolysis capacity of O157 and non-O157 E. coli serotypes from producer-distributor bulk milk

We investigated for virulence genes (stx1, stx2 and hlyA), serotypes and extended-spectrum β-lactamases (ESBLs) producing capacity in O157 and non-O157 Escherichia coli isolated from producer-distributor bulk milk (PDBM). Fifteen different E. coli O-serogroups were observed from the isolates (n = 121). The prevalence of stx1 and stx2 genes among the E. coli isolates was 8.3% and 11.6%, respectively, while 5.8% harboured both stx1 and stx2. Four E. coli isolates (3.3%) had ESBLs producing capacity, resisted multiple cephalosporins and aztreonam, and carried stx genes. Cluster analysis using GTG₅ finger printing revealed a diversity of E. coli seropathotypes in PDBM that are known to be associated with human diarrhoeal diseases. These results highlight a potential risk posed on human health by the consumption of PDBM contaminated with pathogenic E. coli. A further quantitative risk assessment of the impact of pathogenic E. coli contamination in PDBM on human health is therefore recommended.     

Prevalence, serotypes and virulence genes of Shiga toxin-producing Escherichia coli isolated from ovine and caprine milk and other dairy products in Spain

The aim of this study was to determinate the prevalence, serotypes and virulence genes of Shiga toxin-producing Escherichia coli (STEC) strains isolated from different dairy products (DP) in Spain with the purpose of determining whether DP represent a potential source of STEC pathogenic for humans. A total of 502 DP were examined from 64 different ovine and caprine flocks and 6 dairy plants in Extremadura (Western Spain). Samples were collected monthly between March 2003 and June 2004 and included 360 unpasteurised milk obtained from the bulk tank, 103 fresh cheese curds and 39 cheeses. Samples obtained were examined for STEC using genotypic (PCR) methods. STEC strains were detected from 39 (10.8%) bulk tank, 4 (3.9%) fresh cheese curds and 2 (5%) cheese, whereas O157:H7 serotype were isolated from one (0.3%) bulk tank. A total of 9 STEC strains (O27:H18, O45:H38, O76:H19, O91:H28, O157:H7, ONT:H7, ONT:H9 and ONT:H21) were identified in this study. One of them, the serotype O27:H18, has not been reported previously as STEC. PCR showed that 3 strains carried stx1 genes, 5 possessed stx2 genes and 1 both stx1 and stx2. Whereas all STEC caprine isolates showed ehxA genes, only O157:H7 serotype showed eae virulence genes. The strain O157:H7 isolated possessed intimin type gamma1 and belonged to phage type 31. This study confirms that dairy product is an important reservoir of STEC pathogenic for humans.     

Antibacterial activity of oregano and thyme essential oils against Listeria monocytogenes and Escherichia coli O157:H7 in feta cheese packaged under modified atmosphere

The antibacterial activity of the essential oils (EO) of oregano and thyme added at doses of 0.1 or 0.2 and 0.1 ml/100 g, respectively, to feta cheese inoculated with Escherichia coli O157:H7 or Listeria monocytogenes was investigated during cheese storage under modified atmosphere packaging (MAP) of 50% CO₂ and 50% N₂ at 4 °C. Compositional analysis showed that the predominant phenols were carvacrol and thymol for both EO. In control feta inoculated with the pathogens and stored under MAP, results showed that E. coli O157:H7 and L. monocytogenes strains survived up to 32 and 28 days of storage. However, in feta cheese treated with oregano EO at the dose of 0.1 ml/100 g, E. coli O157:H7 or L. monocytogenes survived up to 22 and 18 days, respectively, whereas at the dose of 0.2 ml/100 g up to16 or 14 days, respectively. Feta cheese treated with thyme EO at 0.1 ml/100 g showed populations of E. coli O157:H7 or L. monocytogenes not significantly different (P > 0.05) than those of feta cheese treated with oregano at 0.1 ml/100 g. Although both essential oils exhibited equal antibacterial activity against both pathogens, the populations of L. monocytogenes decreased faster (P < 0.05) than those of E. coli O157:H7 during the refrigerated storage, indicating a stronger antibacterial activity of both essential oils against the former pathogen.     

基于免疫磁分离的7种产志贺毒素大肠埃希氏菌快速检测方法的评价

目的 研究基于免疫磁分离的七种产志贺毒素大肠埃希氏菌快速检测方法的灵敏度与特异性。方法 将大肠埃希氏菌O157：H7和大肠埃希氏菌O103不同稀释度的菌悬液,用免疫磁珠富集后,检测其携带毒力基因stx1、stx2 和黏附基因eae以及O157:H7和O103的抗原基因。同时,对菌悬液进行活菌计数,进行灵敏度研究。对8株携带stx1、stx2、eae基因的目标菌菌悬液,以及25株非目标菌的标准菌株及分离菌株的菌悬液,用免疫磁珠富集后,检测其携带毒力基因stx1、stx2 和黏附基因eae以及抗原基因,进行特异性研究。结果 本方法检测大肠埃希氏菌O157：H7的stx1、stx2、eae以及抗原基因的灵敏度为10_²CFU/mL,检测大肠埃希氏菌O103抗原基因的灵敏度为10_³CFU/mL. 8株目标菌检测结果与其携带的基因一致,没有假阴性,包容性达到100%。25株目标菌检测结果与其携带的基因一致,未发现有假阳性,排他性达到100%。结论 方法具有良好的灵敏性及特异性,适用于食品中七种产志贺毒素大肠埃希氏菌的快速检测。     

Detection and quantitation of enterohemorrhagic Escherichia coli O157, O111, and O26 in beef and bovine feces by real-time polymerase chain reaction

Enterohemorrhagic Escherichia coli (EHEC) O157:H7 and certain non-O157 EHEC serotypes (such as O26:H11, O26: NM, O11:H8, and O111:NM) have emerged as significant causes of human disease throughout the world. Important virulence attributes of EHEC are the intimin protein (encoded by the eae gene) and Shiga toxins 1 and 2 (encoded by the stx1 and stx2 genes, respectively). Two sets of real-time polymerase chain reaction (R-PCR) assays were developed for the simultaneous detection and quantitation of EHEC through the monitoring of the presence of the eae and stx genes, and these assays were evaluated. In the eaeR-PCR assay, three sets of primers and TaqMan probes were designed for the amplification and real-time detection of a portion of the eae gene specific to the EHEC O26, O111, and O157 serotypes. In the stxR-PCR assay, two sets of primers and TaqMan probes were used to amplify and detect the stx1 and stx2 genes. DNA prepared from 67 bacterial strains carrying known virulence markers was tested to determine the specificities of the two assays. In the eaeR-PCR assay, eaeO157- and eaeO111-specific primer-probe sets identified only EHEC O157 and O111 strains, respectively. The eaeO26-specific primer-probe set identified all EHEC 026 isolates and some Shiga toxin-negative serotypes of enteropathogenic E. coli and rabbit diarrheagenic E. coli. The stxR-PCR assay was able to identify only those strains carrying either or both of the Shiga toxin-encoding genes. The detection range of both R-PCR assays was linear over DNA concentrations corresponding to 10(3) to 10(8) CFU/ml of an EHEC strain. Both assays were able to detect and quantify very low levels (1 to 10 CFU/g of food or feces) of EHEC in feces and ground beef enriched for 16 h in a modified Trypticase soy broth. In conclusion, eae- and stx-based R-PCR assays are reliable and sensitive methods for the rapid screening and specific and quantitative detection of important serotypes of EHEC in cattle and in foods of bovine origin.     

牛源性大肠杆菌O157:H7的分离鉴定及其耐酸性比较研究

为比较大肠杆菌（Escherichia coli）O157:H7产毒菌株耐受盐酸和乳酸的差异性,首先采集309 份牛粪及牛肉样,进行菌株分离鉴定,接着采用多重聚合酶链式反应（polymerase chain reaction,PCR）方法检测分离株及其他收集菌株的4 种毒力基因（eae、hly、stx1、stx2）,进而对携带毒力基因的产毒菌株分别进行盐酸和乳酸应激实验。结果表明：共分离鉴定出8 株大肠杆菌O157菌株,样品阳性检出率为2.59%;毒力基因检测表明,8 株菌株均不携带stx1和stx2基因,其中6 株菌株携带eae及hly基因;所有产毒菌株耐酸性实验结果表明,盐酸或乳酸处理2 h后20 株产毒菌株存活菌数均显著下降（P＜0.05）,但下降程度呈现明显的菌株差异性,同一菌株对盐酸、乳酸呈现明显的耐受差异。     

Antagonistic effects of Lactobacillus reuteri against Escherichia coli O157:H7 in white-brined cheese under different storage conditions

This study aimed to investigate the survival of the foodborne pathogen Escherichia coli O157:H7 in white-brined cheeses as influenced by the presence of Lactobacillus reuteri. The white cheeses were made from pasteurized bovine milk inoculated with E. coli O157:H7 (cocktail of 3 strains) to achieve ～5 log₁₀ cfu/g with absence or presence of Lb. reuteri (～6 log₁₀ cfu/g). Cheese samples were brined in 10% or 15% NaCl solution and stored at 10°C and 25°C for 28 d. The white-brined cheeses were assessed for salt content, pH, water activity (A_w), and numbers of E. coli O157:H7, Lb. reuteri, nonstarter lactic acid bacteria (NSLAB), yeasts, and molds. Results showed that E. coli O157:H7 survived in cheese stored in both brine solutions at 10°C and 25°C regardless of the presence of Lb. reuteri. A substantial reduction was observed in cheese stored in 10% NaCl brine at 25°C, followed by cheese stored in 15% NaCl brine at 10°C by 2.64 and 2.16 log₁₀ cfu/g, respectively, in the presence of Lb. reuteri and by 1.02 and 1.87 log₁₀ cfu/g, respectively, in the absence of Lb. reuteri under the same conditions. The pathogen in brine solutions survived but at a lower rate. Furthermore, the growth of Lb. reuteri and NSLAB were enhanced or slightly decreased in cheese and brine by 28 d, respectively. The salt concentrations of cheese ranged from 4 to 6% and 5 to 7% (wt/wt), during 28-d ripening in 10 and 15% brine, respectively. Values of pH and A_w slightly increased at d 1 after exposure to brine and reached 4.69 to 6.08 and 0.91 to 0.95, respectively, in all treatments. Therefore, the addition of Lb. reuteri can be used as a biopreservation method to inhibit the survival of E. coli O157:H7 in white-brined cheese when combined with the appropriate temperature, NaCl level, and storage time.     

Virulence properties of Escherichia coli isolated from Australian dairy powder factory environments

A total of 298 environmental samples from 5 separate dairy powder factories (A–E) were analysed for the presence of Escherichia coli. E. coli was isolated from 80 of these samples to obtain 359 isolates. These isolates were tested for the presence of 12 virulence genes associated with enteropathogenic E. coli (EPEC), atypical EPEC, Shiga toxin (Stx)-producing E. coli (STEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC) and enteroaggregative E. coli (EAEC). A total of 4 isolates obtained from two different sampling points at Factory C were positive for the astA gene, indicative of EAEC. A further 2 isolates from one sample from Factory C were positive for the presence of eaeA and escV genes, indicative of atypical EPEC. No virulence genes were detected in the remaining 353 isolates. While E. coli was present within the environment of dairy powder processing factories the carriage of potential virulence genes was low.     

Molecular detection of nine clinically important non-O157 Escherichia coli serogroups from raw sheep meat in Chaharmahal-va-Bakhtiari province,Iran

STEC isolates and also stx-negative Escherichia coli isolates from sheep meat from the Chaharmahal-va-Bakhtiari province, Iran were analyzed for nine clinically important non-O157 serotypes by PCR. A total of 90 E. coli isolates were tested. Stx-positive and eae-positive E. coli isolates did not belong to the nine most clinically relevant non-O157 STECs. Of the 80 non-STEC isolates, two belonged to the O103 and two belonged to the O128 groups. Stx-negative E. coli O103 and O128 strains isolated have potential in acquiring stx genes and continuing into the digestive system of consumers. Further studies are needed to analyze virulence characteristics of these E. coli strains to determine their potential role in causing disease in humans. For the sake of public health, it is important to monitor and investigate non-O157 diarrheagenic E. coli strains in meat in order to control and prevent them.     

蔬菜中大肠杆菌O157的分离与鉴定

对武汉市售蔬菜(50份)进行大肠杆菌O157的检验,经过新生霉素-EC增菌液增菌、免疫磁珠富集、选择性平板培养和血清学鉴定,从一份生菜中筛出1株O157阳性菌株EC9.23。PCR鉴定该菌毒力基因,stx1、stx2、rfbO157基因均为阳性,hly、eae、fliCH7基因均为阴性。说明从武汉市售蔬菜中可检出携带志贺毒素基因的O157菌株。     

肠出血性大肠杆菌变种的RAPD分型研究

对4株肠出血性大肠杆菌(EHEC)进行随机扩增多态性(RAPD)分析,并结合主要毒力基因如eae和hly,以及志贺毒素滴度,探讨RAPD实验对大肠杆菌致病菌进行基因分型结果的可靠性。实验菌株中有两株变种携带stx基因但不能正常表达志贺毒素,其中一株变种EC169与另两株正常表达志贺毒素的O157菌株具有相似的扩增图谱,而另一株非O157变种EC130与其他菌株聚类明显不同。从20条随机引物中筛选出了重复性强且具有多态性的随机引物G2、G7、G8、G11、G12,可用于大肠杆菌致病菌快速鉴别和食物中毒溯源。