牛源大肠杆菌O157：H7的分离及毒力基因鉴定

从2个牛场采集新鲜粪便,增菌后,免疫磁珠富集,涂布筛选性培养基,挑取可疑菌落用rfbE/fliC二重PCR和血清学方法鉴定。设计毒力基因stx1、stx2、eae、hlyA和tccp相应引物,针对O157：H7对分离株进行PCR鉴定。口服攻毒链霉素处理的BALB/c小鼠明确分离株致病性。结果显示,成功分离到7株出血性大肠杆菌O157：H7,并且有1株迟缓性发酵山梨醇麦康凯培养基。毒力基因检测显示,其中6株毒力因子表型为stx1-stx2＋eae＋hlyA＋tccp＋,另有1株表现型为stx1＋stx2＋eae＋hlyA＋tccp＋,各分离株tccp基因均为阳性,但携带的重复片段数量有差异。所采集样品中肠出血性大肠杆菌O157：H7的检出率高达12%。1×1010 CFU同剂量口服接种经PBS洗涤的5株O157：H7分离株全菌,小鼠存活率有差异分别为40%,50%,60%,20%,50%,各分离株在小鼠体内排菌时间也有差异分别为攻毒后7,9,13,13,15d。     

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.     

Antibiotic‐resistant Shiga toxin‐producing Escherichia coli: An overview of prevalence and intervention strategies

Shiga toxin‐producing Escherichia coli (STEC) are foodborne pathogens that can cause severe diseases, including bloody diarrhoea and kidney failure, in humans, while remaining harmless to its primary reservoir hosts, cattle. Antibiotics such as azithromycin, fosfomycin and meropenem are being used and recommended in the treatment of early‐stage STEC (mainly E. coli O157:H7) infections, as these are reportedly effective in preventing Shiga toxin release and kidney failure while eliminating the pathogen. However, antibiotic resistance among STEC isolates could negatively impact these and other similar treatment options while contributing towards the spread of antibiotic resistance genes especially if encoded on mobile genetic elements like plasmids. Antibiotic resistance among STEC isolates recovered from animals and patients is being reported globally. A comprehensive understanding of the prevalence of antibiotic‐resistant STEC (AR‐STEC) and the mechanisms promoting this resistance among these bacteria could help direct therapies and develop strategies to effectively reduce/eliminate these pathogens. Here, we have reviewed literature from the past three decades to gain insights on this prevalence and its impact on human infections. In addition, we have reviewed various strategies proposed by researchers to control STEC that in turn would be applicable to AR‐STEC as well.     

Phage and MLVA Typing of Salmonella Enteritidis Isolated from Layers and Humans in Belgium from 2000–2010, A Period in which Vaccination of Laying Hens was Introduced

The aim of the study was to characterize isolates of Salmonella enterica serovar Enteritidis (S. Enteritidis) obtained from humans and layer farms in Belgium collected during 2000–2010. Three periods were compared, namely (i) before implementation of vaccination (2000–2004), (ii) during voluntary vaccination (2005–2006) and (iii) during implementation of the national control program (NCP) for Salmonella including mandatory vaccination against S. Enteritidis (2007–2010). The characteristics compared across time periods were distributions of phage type and multiple‐locus variable number tandem‐repeat assay (MLVA). While PT4 and PT21 were predominantly isolated in Belgium in layers and humans before 2007, a significant reduction of those PTs was observed in both populations in the period 2007–2010. The relative proportion of PT4b, PT21c and PT6c was found to have increased considerably in the layer population since 2007. In the human population, PT8, PT1 and the group of ‘other’ PTs were more frequently isolated compared to the previous periods. When comparing the proportion of the predominant MLVA types Q2 and U2, no significant difference was found between the layer and human population in the three periods and between periods within each category (layer and human). A significant difference in isolate distribution among MLVA clusters I and II was found between human and layer isolates recovered during Period 3 and in the human population between Period 1 and 3. Results suggest that the association between S. Enteritidis in layers and the occurrence of the pathogen in humans changed since implementation of the NCP in 2007.     

Isolation and characterisation of Shiga toxin-producing Escherichia coli O157:H7 from calves in Argentina

Four hundred and twenty-two calves were examined for intestinal carriage of Shiga toxin-producing Escherichia coli O157:H7 using conventional plating. Two (0.5%) E. coli O157 were recovered. They were compared with 96 Argentine strains of different origin by pulsed-field gel electrophoresis, phage typing and PCR-RFLP of stx2 genes. One strain isolated from a calf, was closely related with 18 strains of clinical origin.     

An outbreak of Shiga toxin‐producing Escherichia coli O157:H7 linked to a mud‐based obstacle course,England, August 2018

In August 2018, Public Health England (PHE) was made aware of five probable cases of Shiga toxin‐producing Escherichia coli (STEC) O157:H7 among individuals reporting participation in a mud‐based obstacle race. An additional four cases, identified via routine whole‐genome sequencing, were subsequently linked to the same event. Two of the nine cases were due to secondary household transmission. Despite an agreement between the event organizers and the local authority, to ensure that all livestock were removed from the site 28 days before the event, sheep were observed grazing on some of the routes taken by the runners 2 days prior to the race taking place. A retrospective review of incidents reported to PHE between 2015 and 2018 identified 41 cases of gastroenteritis associated with muddy assault course events. Of these, 25 cases were due to infection with STEC O157:H7, of which all but one were associated with outbreaks. Due to the environment in which such events take place, it is impossible to entirely remove the risk of exposure to potentially pathogenic zoonoses. However, race organizers should ensure that livestock are removed from the course 28 days before the event. They should also ensure that participants are made aware of the risk of contracting gastrointestinal disease from the environment, and to stress the importance of hand hygiene post‐event and the risk of secondary transmission, particularly to children who are at risk of developing haemolytic uraemic syndrome.     

Assessing the Existing Information on the Efficacy of Bovine Vaccination against Escherichia coli O157:H7 – A Systematic Review and Meta‐analysis

The objective of this study was to conduct a systematic review and meta‐analysis to evaluate the existing information on the efficacy of commercial vaccination to reduce the prevalence of Escherichia coli O157:H7 in weaned cattle in beef feedlot finishing systems under commercial conditions. Currently, only two commercial vaccines exist, and thus, only publications reporting the use of vaccines targeting type III secreted proteins and/or siderophore receptor and porin proteins (SRP) were considered relevant. A total of 18 studies reporting 45 comparisons were included in this review. Meta‐analyses were conducted variously on (i) pre‐harvest outcomes, (ii) at‐harvest outcomes and (iii) both pre‐harvest and at‐harvest outcomes combined. Overall, efficacy of vaccination was consistently observed. Efficacy and homogeneity of the results was demonstrated for the two‐dose regimen, allowing us to conclude with confidence that the two‐dose approach is efficacious. For pre‐harvest outcomes and two‐dose regimens, the odds ratios (OR) were 0.53 (95% CI = 0.45–0.62) for the two vaccines combined and 0.49 (95% CI = 0.40–0.60) for vaccine targeting type III secreted proteins. The test for heterogeneity among studies yielded a Q test P = 0.354 for the two vaccines combined and Q test P = 0.269 for the vaccine targeting type III secreted proteins, indicating homogeneity in both cases. For pre‐ and at‐harvest outcomes combined and two‐dose regimens, the odds ratios (OR) were 0.52 (95% CI = 0.44–0.61) for the two vaccines combined and 0.45 (95% CI = 0.34–0.60) for vaccine targeting type III secreted proteins. The test for heterogeneity among studies yielded a Q test P = 0.134 for the two vaccines combined indicating homogeneity and Q test P = 0.089 for the vaccine targeting type III secreted proteins indicating heterogeneity. Based on this meta‐analysis, bovine vaccination appears to be an effective approach to the pre‐harvest control of E. coli O157:H7.     

Genetic Variations in Shiga Toxin‐Producing Abilities of Bovine and Human Escherichia coli O157:H7

Cattle are a primary reservoir of Escherichia coli O157:H7, a major foodborne pathogen. The organism causes haemorrhagic colitis which can lead to serious complications, including haemolytic–uraemic syndrome. Although E. coli O157:H7 is widely prevalent in cattle and cattle environments, the number of human cases remain relatively low, suggesting possible strain diversity and differences in virulence between human and bovine strains. Shiga toxins, Stx1 and Stx2, are the major virulence factors. Differences in Stx2 production between human and bovine strains have been demonstrated previously, and isolates possessing the stx₂ gene, but not producing Stx2 [toxin non‐producing (TNP) strains] have been identified. In this study, 150 isolates (56 human, 94 bovine) were tested by PCR for stx2 upstream regions associated with TNP and the Q933 gene, which has been previously associated with toxin production. A reverse passive latex agglutination test was used to evaluate 107 isolates (50 human, 57 bovine) for Stx1 and Stx2 production. The percentages of human and bovine isolates positive for presence of the TNP regions were similar (57.1% and 53.1% respectively), while a higher percentage of human isolates was positive for Q933 gene (89.3% versus 54.3%). Stx2 production of ≥1 : 8 was found in 86.0% of human isolates compared with 26.3% of bovine isolates. Bovine isolates with the presence of the TNP regions were associated with significantly lower Stx2 production (P < 0.05), while the Q933 gene was associated with higher Stx2 production (P < 0.05). However, the presence of the TNP region was not associated (P > 0.05) with low Stx2 production in human isolates. Therefore, Q933 was a better indicator of high Stx2 production by human and bovine isolates and may be a useful screening method to assess their potential to cause human disease.     

Genetic diversity,anti‐microbial resistance,plasmid profile and frequency of the Vi antigen in Salmonella Dublin strains isolated in Brazil

Salmonella Dublin is strongly adapted to cattle causing enteritis and/or systemic disease with high rates of mortality. However, it can be sporadically isolated from humans, usually causing serious disease, especially in patients with underlying chronic diseases. The aim of this study was to molecularly type S. Dublin strains isolated from humans and animals in Brazil to verify the diversity of these strains as well as to ascertain possible differences between strains isolated from humans and animals. Moreover, the presence of the capsular antigen Vi and the plasmid profile was characterized in addition to the anti‐microbial resistance against 15 drugs. For this reason, 113 S. Dublin strains isolated between 1983 and 2016 from humans (83) and animals (30) in Brazil were typed by PFGE and MLVA. The presence of the capsular antigen Vi was verified by PCR, and the phenotypic expression of the capsular antigen was determined serologically. Also, a plasmid analysis for each strain was carried out. The strains studied were divided into 35 different PFGE types and 89 MLVA‐types with a similarity of ≥80% and ≥17.5%, respectively. The plasmid sizes found ranged from 2 to >150 kb and none of the strains studied presented the capsular antigen Vi. Resistance or intermediate resistance was found in 23 strains (20.3%) that were resistant to ampicillin, ciprofloxacin, chloramphenicol, imipenem, nalidixic acid, piperacillin, streptomycin and/or tetracycline. The majority of the S. Dublin strains studied and isolated over a 33‐year period may descend from a common subtype that has been contaminating humans and animals in Brazil and able to cause invasive disease even in the absence of the capsular antigen. The higher diversity of resistance phenotypes in human isolates, as compared with animal strains, may be a reflection of the different anti‐microbial treatments used to control S. Dublin infections in humans in Brazil.     

Molecular Typing of Mycobacterium bovis Isolates in Argentina: First Description of a Person‐to‐Person Transmission Case

Bovine tuberculosis is caused by Mycobacterium bovis, a mycobacterium highly similar to M. tuberculosis that belongs to the M. tuberculosis complex. The main host of M. bovis is cattle but it also affects many other mammalians including humans. Tuberculosis in humans caused by either M. bovis or M. tuberculosis is clinically hard to distinguish. During 2004–2005, samples from 448 patients with diagnosis of TB were collected from different regions of Argentina. The PRA technique identified 400 isolates with representative patterns of mycobacterium. The predominant ones were the M. tuberculosis complex, the M. avium–M. intracellulare complex and M. gordonae. Samples with M. tuberculosis complex PRA restriction profiles were analyzed with a multiplex PCR to differentiate between M. tuberculosis and M. bovis. Multiplex PCR identified nine M. bovis. The results allowed the possibility to establish that 2% of pulmonary tuberculosis was due to M. bovis. Isolates of M. bovis from humans were examined using spoligotyping. These isolates presented five different spoligotypes. The main spoligotype was also the most frequently one found in cattle. The remaining human spoligotypes (grouped in clusters) are occasionally found in cattle. Variable number tandem repeat (VNTR) analysis identified five different patterns. By combining the results of spoligotyping and VNTR analysis, we were able to differentiate seven M. bovis isolates. The remaining two M. bovis samples showed the same spoligotype and VNTR profile and belonged to household contacts. An MDR‐M. bovis was isolated from the samples of these household contacts. The identification of two epidemiologically linked cases of human M. bovis infection suggests person‐to‐person transmission of an MDR‐M. bovis.     

Molecular characterization of Escherichia coli O157:H7 strains isolated from different sources and geographic regions

Escherichia (E.) coli serotype O157:H7 is a globally distributed human enteropathogen and is comprised of microorganisms with closely related genotypes. The main reservoir for this group is bovine bowels, and infection mainly occurs after ingestion of contaminated water and food. Virulence genetic markers of 28 O157:H7 strains were investigated and multilocus enzyme electrophoresis (MLEE) was used to evaluate the clonal structure. O157:H7 strains from several countries were isolated from food, human and bovine feces. According to MLEE, O157:H7 strains clustered into two main clonal groups designated A and B. Subcluster A1 included 82% of the O157:H7 strains exhibiting identical MLEE pattern. Most enterohemorrhagic E. coli (EHEC) O157:H7 strains from Brazil and Argentina were in the same MLEE subgroup. Bovine and food strains carried virulence genes associated with EHEC pathogenicity in humans.     

新疆牛源大肠杆菌O157∶H7的分离鉴定及ERIC-PCR基因分型研究

为了了解牛源大肠杆菌（E.coli）O157∶H7在新疆地区的污染状况以及遗传多样性,探究不同地区分离菌株的遗传关系,为控制牛源E.coli O157∶H7的传播提供试验依据。将采集的样品在EC肉汤中进行增菌（37 ℃、180 r/min）,接着将增菌液划线接种到SMAC平板上,37 ℃培养箱中过夜培养18 h左右。挑取SMAC平板上白色或无色单菌落接种MUG培养基,37 ℃培养18 h左右,将无荧光样品接种到SMAC平板上,37 ℃培养18 h左右,隔天挑取白色或无色单菌落进行PCR鉴定,具有rfbE和fliC基因条带的即为阳性菌株。将阳性菌株进行肠杆菌基因间重复共有序列扩增（ERIC-PCR）指纹图谱聚类分析,分析菌株之间的同源性关系。ERIC-PCR结果显示,相似性100%的菌株有3组。从伊犁地区分离到的菌株差异性最大,具有6种分型;其次是乌鲁木齐,具有4种分型。菌株来源多样性最多的在D簇,由此可见通过ERIC-PCR分型,可以进行溯源观察。ERIC-PCR能够区分特定采样点或物种的分离物,它能够证明从不同来源的菌株之间,存在着某些相似的ERIC特性,并聚集在同一个簇群中。该研究中筛选出的E.coli O157∶H7菌株具有广泛的遗传多样性,该方法对于检测不同物种间的细菌差异非常敏感。由此可见ERIC-PCR可以作为E.coli O157∶H7常规监测和鉴定的一个有效的工具。     

An investigation of shedding and super‐shedding of Shiga toxigenic Escherichia coli O157 and E. coli O26 in cattle presented for slaughter in the Republic of Ireland

Shiga toxigenic Escherichia coli (STEC) are an important group of pathogens and can be transmitted to humans from direct or indirect contact with cattle faeces. This study investigated the shedding of E. coli O157 and O26 in cattle at the time of slaughter and factors associated with super‐shedding (SS) animals. Rectoanal mucosal swab (RAMS) samples were collected from cattle (n = 1,317) at three large Irish commercial beef abattoirs over an 18 month period, and metadata were collected at the time of sampling regarding farm of origin, animal age, breed and gender. RAMS swabs were examined for the presence and numbers of E. coli O157 and O26 using a previously developed quantitative real‐time PCR protocol. Samples positive by PCR were culturally examined and isolates analysed for the presence of stx subtypes, eae and phylogroup. Any samples with counts >10⁴ CFU/swab of STEC O157 or O26 were deemed to be super‐shedders. Overall, 4.18% (55/1,317) of RAMS samples were positive for STEC O157, and 2.13% (28/1,317) were classified as STEC O157 SS. For STEC O26, 0.76% (10/1,317) of cattle were positive for STEC O26, and 0.23% (3/1,317) were classified as super‐shedders. Fewer STEC shedders and SS were noted among older animals (>37 months). There was a seasonal trend observed for STEC O157, with the highest prevalence of shedding and SS events in the autumn (August to October). The majority of E. coli O157 (50/55) isolates had stx2 and were eae positive, with no significant difference between SS and low shedders (LS). Interestingly, all STEC O26 (n = 10) were eae negative and had varied stx profiles. This study demonstrates that, while the overall shedding rates are relatively low in cattle at slaughter, among positive animals there is a high level of SS, which may pose a higher risk of cross‐contamination during slaughter.     

牛源大肠杆菌O157:H7的分离鉴定

为调查新疆部分地区E.coli O157：H7的感染情况和菌株致病性,从新疆阿克苏、伊犁、塔城3个地区的牛场采集新鲜粪样564份,对E.coli O157：H7进行分离与鉴定。利用E.coli营养肉汤（EC肉汤）对样品进行增菌后,用山梨醇麦康凯培养基（SMAC）平板选择性培养,再经过4-甲基伞形酮-β-D葡萄糖醛酸苷培养基（MUG）的筛选,对疑似菌株进行生化和PCR鉴定,并将分离鉴定到的菌株进行小鼠攻毒试验。结果显示,从伊犁地区采集的样品中共分离出2株E.coli O157：H7（Y166和Y226）,其检出率为0.88%;小鼠攻毒试验中,Y166和Y226试验组小鼠在48 h内全部死亡,具有一定致病性;从阿克苏、塔城所采样品中未分离到E.coli O157：H7。     

The Use of Direct‐Fed Microbials to Reduce Shedding of Escherichia coli O157 in Beef Cattle: A Systematic Review and Meta‐analysis

Human illness due to infections with Escherichia coli O157 is a serious health concern. Infection occurs through direct contact with infected animals or their faeces, through contaminated food or water and/or through person‐to‐person transmission. A reduction in faecal E. coli O157 shedding in cattle might reduce the burden of human infections. We used systematic review and meta‐analysis to assess the efficacy of direct‐fed microbials (DFM), compared with placebo or no treatment, fed during the pre‐harvest stage of production in reducing faecal E. coli O157 shedding in beef cattle during field trials. Four electronic databases, Nebraska Beef Reports and review article reference lists were searched. A total of 16 publications assessing faecal shedding at the end of the trial and/or throughout the trial period were included. The majority of publicly disseminated trials evaluated the prevalence of E. coli O157 faecal shedding; only two evaluated the concentration of organisms in faeces. The prevalence of faecal E. coli O157 shedding in cattle is significantly reduced by DFM treatments (summary effect size for all DFM – OR = 0.46; CI = 0.36–0.60). The DFM combination Lactobacillus acidophilus (NP51) and Propionibacterium freudenreichii (NP24) was more efficacious in reducing the prevalence of faecal E. coli O157 shedding at the time of harvest and throughout the trial period compared with the group of other DFM, although this difference was not statistically significant. Furthermore, we found that the combination [NP51 and NP24] treatment was more efficacious in reducing the prevalence of faecal E. coli O157 shedding at the time of harvest and throughout the trial period when fed at the dose of 10⁹ CFU/animal/day than any lesser amount, although this difference was not statistically significant. Feeding beef cattle DFM during the pre‐harvest stage of production reduces the prevalence of E. coli O157 faecal shedding and might effectively reduce human infections.     

Spatial Epidemiology of Escherichia coli O157:H7 in Dairy Cattle in Relation to Night Roosts Of Sturnus vulgaris (European Starling) in Ohio,USA (2007–2009)

The goal of our study was to use spatial scan statics to determine whether the night roosts of European starlings (Sturnus vulgaris) act as point sources for the dissemination of Escherichia coli O157:H7 among dairy farms. From 2007 to 2009, we collected bovine faecal samples (n = 9000) and starling gastrointestinal contents (n = 430) from 150 dairy farms in northeastern Ohio, USA. Isolates of E. coli O157:H7 recovered from these samples were subtyped using multilocus variable‐number tandem repeat analysis (MLVA). Generated MLVA types were used to construct a dendrogram based on a categorical multistate coefficient and unweighted pair‐group method with arithmetic mean (UPGMA). Using a focused spatial scan statistic, we identified statistically significant spatial clusters among dairy farms surrounding starling night roosts, with an increased prevalence of E. coli O157:H7‐positive bovine faecal pats, increased diversity of distinguishable MLVA types and a greater number of isolates with MLVA types from bovine‐starling clades versus bovine‐only clades. Thus, our findings are compatible with the hypothesis that starlings have a role in the dissemination of E. coli O157:H7 among dairy farms, and further research into starling management is warranted.     

Detection of Escherichia coli O157 and Escherichia coli O157:H7 by the immunomagnetic separation technique and stx1 and stx2 genes by multiplex PCR in slaughtered cattle in Samsun Province, Turkey

This study was conducted to investigate the presence of Escherichia (E.) coli O157 and E. coli O157:H7 and stx1 and stx2 genes on cattle carcasses and in rectal samples collected from Samsun Province of Turkey. A total of 200 samples collected from cattle carcasses and the rectal contents of 100 slaughtered cattle from two commercial abattoirs were tested using the immunomagnetic separation technique and multiplex PCR methods. E. coli O157 and E. coli O157:H7 were detected in 52 of the 200 samples (26%) tested. Of the positive samples, 49 were E. coli O157 and three were E. coli O157:H7. The E. coli O157 strain was isolated from 24 carcasses and 25 rectal samples, while E. coli O157:H7 was isolated from two carcasses and one rectal sample. Of the 49 samples positive for E. coli O157, 32 were from the rectal and carcass samples of the same animal, while two E. coli O157:H7 isolates were obtained from rectal swabs and carcasses of the same animal. The stx1 and stx2 genes were both detected in 35 E. coli O157 isolates and one E. coli O157:H7 isolate, but the stx2 gene was only detected alone in two E. coli O157 isolates. Overall, 16 carcasses tested positive for E. coli O157 and one carcass tested positive for E. coli O157:H7 based on both carcass and rectal samples. Overall, the results of this study indicate that cattle carcasses pose a potential risk to human health due to contamination by E. coli O157 and E. coli O157:H7 in the feces.     

Spatio‐Temporal Scan Statistics for the Detection of Outbreaks Involving Common Molecular Subtypes: Using Human Cases of Escherichia coli O157:H7 Provincial PFGE Pattern 8 (National Designation ECXAI.0001) in Alberta as an Example

Molecular typing methods have become a common part of the surveillance of foodborne pathogens. In particular, pulsed‐field gel electrophoresis (PFGE) has been used successfully to identify outbreaks of Escherichia coli O157:H7 in humans from a variety of food and environmental sources. However, some PFGE patterns appear commonly in surveillance systems, making it more difficult to distinguish between outbreak and sporadic cases based on molecular data alone. In addition, it is unknown whether these common patterns might have unique epidemiological characteristics reflected in their spatial and temporal distributions. Using E. coli O157:H7 surveillance data from Alberta, collected from 2000 to 2002, we investigated whether E. coli O157:H7 with provincial PFGE pattern 8 (national designation ECXAI.0001) clustered in space, time and space–time relative to other PFGE patterns using the spatial scan statistic. Based on our purely spatial and temporal scans using a Bernoulli model, there did not appear to be strong evidence that isolates of E. coli O157:H7 with provincial PFGE pattern 8 are distributed differently from other PFGE patterns. However, we did identify space–time clusters of isolates with PFGE pattern 8, using a Bernoulli model and a space–time permutation model, which included known outbreaks and potentially unrecognized outbreaks or additional outbreak cases. There were differences between the two models in the space–time clusters identified, which suggests that the use of both models could increase the sensitivity of a quantitative surveillance system for identifying outbreaks involving isolates sharing a common PFGE pattern.