non-O157 Shiga Toxin-producing Escherichia Coli Infections Research Articles

Risk Factors for Non-O157 Shiga Toxin–Producing Escherichia coli Infections, United States

Shiga toxin-producing Escherichia coli (STEC) causes acute diarrheal illness. To determine risk factors for non-O157 STEC infection, we enrolled 939 patients and 2,464 healthy controls in a case-control study conducted in 10 US sites. The highest population-attributable fractions for domestically acquired infections were for eating lettuce (39%), tomatoes (21%), or at a fast-food restaurant (23%). Exposures with 10%-19% population attributable fractions included eating at a table service restaurant, eating watermelon, eating chicken, pork, beef, or iceberg lettuce prepared in a restaurant, eating exotic fruit, taking acid-reducing medication, and living or working on or visiting a farm. Significant exposures with high individual-level risk (odds ratio >10) among those >1 year of age who did not travel internationally were all from farm animal environments. To markedly decrease the number of STEC-related illnesses, prevention measures should focus on decreasing contamination of produce and improving the safety of foods prepared in restaurants.

Multiple-Locus Variable-Number Tandem Repeat Analysis Scheme for Non-O157 Shiga Toxin-Producing Escherichia coli: Focus on Serogroups O103, O121, O145, O165, and O91.

Non-O157 Shiga toxin-producing Escherichia coli (STEC) infections are a growing concern for public health. The number of sporadic cases and outbreaks of non-O157 STEC infections have increased in recent years. Molecular subtyping is an essential tool that allows high-resolution and rapid differentiation of isolates, identification of case clusters, and detection of outbreak clusters. Multiple-locus variable-number tandem repeat analysis (MLVA) is one of the most useful typing methods for differentiating isolates that cause foodborne diseases. In Japan, serogroups O26, O111, O103, O121, O145, O165, and O91 have been frequently isolated or associated with severe cases of non-O157 STEC infections. In this study, we designed an MLVA scheme (MLVA43) for serogroups O103, O121, O145, O165, and O91 by adding 26 new loci to an MLVA scheme (MLVA17) previously developed by our group for serogroups O157, O26, and O111 using 17 loci. We found that the discriminatory power of MLVA43 was comparable to that of pulsed-field gel electrophoresis (PFGE) for serogroups O103, O145, O165, and O91, and superior to that of PFGE for O121. MLVA43 identified more profiles than did MLVA17, except for serogroup O111 with 707 isolates. The MLVA43 scheme will enable rapid detection of outbreak clusters, which will aid in implementing rapid control measures against non-O157 STEC infections.

DNA Microarray-Based Genomic Characterization of the Pathotypes of Escherichia coli O26, O45, O103, O111, and O145 Isolated from Feces of Feedlot Cattle

Shiga toxin-producing Escherichia coli (STEC) serogroups O26, O45, O103, O111, O121, and O145, referred to as the top six non-O157 serogroups, are responsible for more than 70% of human non-O157 STEC infections in North America. Cattle harbor non-O157 strains in the hindgut and shed them in the feces. The objective of this study was to use the U.S. Food and Drug Administration (FDA) E. coli identification (ECID) DNA microarray to identify the serotype, assess the virulence potential of each, and determine the phylogenetic relationships among five of the six non-O157 E. coli serogroups isolated from feedlot cattle feces. Forty-four strains of STEC, enterohemorrhagic E. coli (EHEC), enteropathogenic E. coli (EPEC), or putative nonpathotype E. coli (NPEC) of cattle origin and five human clinical strains of EHEC were assayed with the FDA-ECID DNA microarray. The cattle strains harbored diverse flagellar genes. The bovine and human strains belonging to serogroups O26, O45, and O103 carried stx1 only, O111 carried both stx1 and stx2, and O145 carried either stx1 or stx2. The strains were also positive for various subtypes of intimin and other adhesins (IrgA homologue adhesin, long polar fimbriae, mannose-specific adhesin, and curli). Both human and cattle strains were positive for LEE-encoded type III secretory system genes and non-LEE-encoded effector genes. SplitsTree4, a program used to determine the phylogenetic relationship among the strains, revealed that the strains within each serogroup clustered according to their pathotype. In addition to genes encoding Shiga toxins, bovine non-O157 E. coli strains possessed other major virulence genes, including those for adhesins, type III secretory system proteins, and plasmid-borne virulence genes, similar to human clinical strains. Because virulence factors encoded by these genes are involved in the pathogenesis of various pathotypes of E. coli, the bovine non-O157 strains could cause human illness. The FDA-ECID DNA microarray assay rapidly provided a profile of the virulence genes for assessment of the virulence potential of each strain.

Genetic diversity of the enterohaemolysin gene (ehxA) in non-O157 Shiga toxin-producing Escherichia coli strains in China

Non-O157 Shiga toxin-producing Escherichia coli (STEC) is increasingly recognized as an important enteric foodborne pathogen. The hallmark of the disease is the production of Shiga toxins; however, there are other virulence factors that contribute to the pathogenesis of STEC. This study aimed to investigate the prevalence and genetic diversity of the enterohaemolysin gene, ehxA, among non-O157 STEC strains from human, animal, and food sources. The ehxA gene was amplified from 138 (31.8%) of 434 non-O157 STEC strains, among which 36 unique ehxA sequences were identified. Based on ehxA sequence analysis, three phylogenetic ehxA groups (I II, and III) were determined. Correlations between ehxA groups and sources, serotypes, and virulent gene profiles were observed. The ehxA group II strains were mostly diarrhoeal patient-derived and may demonstrate higher pathogenic potential compared with the ehxA group I and group III strains. Five types of replicons (I1-Ig, FIB, K, F, and B/O) were identified in the 138 ehxA-positive strains, and 3.6%, 5.8%, and 52.2% of the strains harboured toxB, katP and espP genes, respectively, implying marked genetic diversity of ehxA containing plasmids in non-O157 STEC strains. Sequence-based ehxA genotyping might be important in modern strain typing and in epidemiological surveillance of non-O157 STEC infections.

Detection, Characterization, and Typing of Shiga Toxin-Producing Escherichia coli.

Shiga toxin-producing Escherichia coli (STEC) are responsible for gastrointestinal diseases reported in numerous outbreaks around the world. Given the public health importance of STEC, effective detection, characterization and typing is critical to any medical laboratory system. While non-O157 serotypes account for the majority of STEC infections, frontline microbiology laboratories may only screen for STEC using O157-specific agar-based methods. As a result, non-O157 STEC infections are significantly under-reported. This review discusses recent advances on the detection, characterization and typing of STEC with emphasis on work performed at the Alberta Provincial Laboratory for Public Health (ProvLab). Candidates for the detection of all STEC serotypes include chromogenic agars, enzyme immunoassays (EIA) and quantitative real time polymerase chain reaction (qPCR). Culture methods allow further characterization of isolates, whereas qPCR provides the greatest sensitivity and specificity, followed by EIA. The virulence gene profiles using PCR arrays and stx gene subtypes can subsequently be determined. Different non-O157 serotypes exhibit markedly different virulence gene profiles and a greater prevalence of stx1 than stx2 subtypes compared to O157:H7 isolates. Finally, recent innovations in whole genome sequencing (WGS) have allowed it to emerge as a candidate for the characterization and typing of STEC in diagnostic surveillance isolates. Methods of whole genome analysis such as single nucleotide polymorphisms and k-mer analysis are concordant with epidemiological data and standard typing methods, such as pulsed-field gel electrophoresis and multiple-locus variable number tandem repeat analysis while offering additional strain differentiation. Together these findings highlight improved strategies for STEC detection using currently available systems and the development of novel approaches for future surveillance.

Epidemiology and microbiology of Shiga toxin-producing Escherichia coli other than serogroup O157 in England, 2009-2013.

The implementation of direct testing of clinical faecal specimens for gastrointestinal (GI) pathogens by PCR offers a sensitive and comprehensive approach for the detection of Shiga toxin-producing Escherichia coli (STEC). The introduction of a commercial PCR assay, known as GI PCR, for the detection of GI pathogens at three frontline hospital laboratories in England between December 2012 and December 2013 led to a significant increase in detection of STEC other than serogroup O157 (non-O157 STEC). In 2013, 47 isolates were detected in England, compared with 57 in the preceding 4 years (2009-2012). The most common non-O157 STEC serogroup detected was O26 (23.2 %). A total of 47 (47.5 %) STEC isolates had stx2 only, 28 (28.3 %) carried stx1 and stx2, and the remaining 24 (24.2 %) had stx1 only. Stx2a (64.0 %) was the most frequently detected Stx2 subtype. The eae (intimin) gene was detected in 52 (52.5 %) non-O157 STEC isolates. Six strains of STEC O104 had aggR, but this gene was not detected in any other STEC serogroups in this study. Haemolytic ureamic syndrome was significantly associated with STEC strains possessing eae [odds ratio (OR) 5.845, P = 0.0235] and/or stx2a (OR 9.56, P = 0.0034) subtypes. A matched case-control analysis indicated an association between non-O157 STEC cases and contact with farm animals. Widespread implementation of the PCR approach in England will determine the true incidence of non-O157 STEC infection, highlight the burden in terms of morbidity and mortality, and facilitate the examination of risk factors to indicate whether there are niche risk exposures for particular strains.

Multiple‐Aetiology Enteric Infections Involving Non‐O157 Shiga Toxin‐Producing Escherichia coli – FoodNet, 2001–2010

We describe multiple-aetiology infections involving non-O157 Shiga toxin-producing Escherichia coli (STEC) identified through laboratory-based surveillance in nine FoodNet sites from 2001 to 2010. A multiple-aetiology infection (MEI) was defined as isolation of non-O157 STEC and laboratory evidence of any of the other nine pathogens under surveillance or isolation of >1 non-O157 STEC serogroup from the same person within a 7-day period. We compared exposures of patients with MEI during 2001-2010 with those of patients with single-aetiology non-O157 STEC infections (SEI) during 2008-2009 and with those of the FoodNet population from a survey conducted during 2006-2007. In total, 1870 non-O157 STEC infections were reported; 68 (3.6%) were MEI; 60 included pathogens other than non-O157 STEC; and eight involved >1 serogroup of non-O157 STEC. Of the 68 MEI, 21 (31%) were part of six outbreaks. STEC O111 was isolated in 44% of all MEI. Of patients with MEI, 50% had contact with farm animals compared with 29% (P < 0.01) of persons with SEI; this difference was driven by infections involving STEC O111. More patients with non-outbreak-associated MEI reported drinking well water (62%) than respondents in a population survey (19%) (P < 0.01). Drinking well water and having contact with animals may be important exposures for MEI, especially those involving STEC O111.

Characterization of Shiga toxin – producing Escherichia coli infections in beef feeder calves and the effectiveness of a prebiotic in alleviating Shiga toxin - producing Escherichia coli infections

BackgroundIn the less-sensitive mouse model, Shiga toxin-producing Escherichia coli (STEC) challenges result in shedding that reflect the amount of infection and the expression of virulence factors such as Shiga toxins (Stx). The purpose of this study was to characterize the contribution of STEC diversity and Stx expression to shedding in beef feeder calves and to evaluate the effectiveness of a prebiotic, Celmanax®, to alleviate STEC shedding. Fecal samples were collected from calves at entry and after 35 days in the feedlot in spring and summer. STECs were evaluated using selective media, biochemical profile, serotyping and Stx detection. Statistical analysis was performed using repeated measures ANOVA and logistic regression.ResultsAt entry, non-O157 STEC were dominant in shedding calves. In spring, 21%, 14% and 14% of calves acquired O157, non-O157 and mixed STEC infections, respectively. In contrast, 45%, 48% and 46% of calves in summer acquired O157, non-O157 and mixed STEC infections, respectively. Treatment with a prebiotic, Celmanax®, in spring significantly reduced 50% of the O157 STEC infections, 50% of the non-O157 STEC infections and 36% of the STEC co-infections (P = 0.037). In summer, there was no significant effect of the prebiotic on STEC infections. The amount of shedding at entry was significantly related to the number and type of STECs present and Stx expression (r2 = 0.82). The same relationship was found for shedding at day 35 (r2 = 0.85), but it was also related to the number and type of STECs present at entry. Stx - producing STEC infections resulted in 100 to 1000 × higher shedding in calves compared with Stx-negative STECs.ConclusionsSTEC infections in beef feeder calves reflect the number and type of STECs involved in the infection and STEC expression of Stx. Application of Celmanax® reduced O157 and non-O157 STEC shedding by calves but further research is required to determine appropriate dosages to manage STEC infections.

Multistate Outbreak of Escherichia coli O145 Infections Associated with Romaine Lettuce Consumption, 2010

Non-O157 Shiga toxin–producing Escherichia coli (STEC) can cause severe illness, including hemolytic uremic syndrome (HUS). STEC O145 is the sixth most commonly reported non-O157 STEC in the United States, although outbreaks have been infrequent. In April and May 2010, we investigated a multistate outbreak of STEC O145 infection. Confirmed cases were STEC O145 infections with isolate pulsed-field gel electrophoresis patterns indistinguishable from those of the outbreak strain. Probable cases were STEC O145 infections or HUS in persons who were epidemiologically linked. Case-control studies were conducted in Michigan and Ohio; food exposures were analyzed at the restaurant, menu, and ingredient level. Environmental inspections were conducted in implicated food establishments, and food samples were collected and tested. To characterize clinical findings associated with infections, we conducted a chart review for case patients who sought medical care. We identified 27 confirmed and 4 probable cases from five states. Of these, 14 (45%) were hospitalized, 3 (10%) developed HUS, and none died. Among two case-control studies conducted, illness was significantly associated with consumption of shredded romaine lettuce in Michigan (odds ratio [OR] = undefined; 95% confidence interval [CI] = 1.6 to undefined) and Ohio (OR = 10.9; 95% CI = 3.1to 40.5). Samples from an unopened bag of shredded romaine lettuce yielded the predominant outbreak strain. Of 15 case patients included in the chart review, 14 (93%) had diarrhea and abdominal cramps and 11 (73%) developed bloody diarrhea. This report documents the first foodborne outbreak of STEC O145 infections in the United States. Current surveillance efforts focus primarily on E. coli O157 infections; however, non-O157 STEC can cause similar disease and outbreaks, and efforts should be made to identify both O157 and non-O157 STEC infections. Providers should test all patients with bloody diarrhea for both non-O157 and O157 STEC.

Increased Recognition of Non-O157 Shiga Toxin–ProducingEscherichia coliInfections in the United States During 2000–2010: Epidemiologic Features and Comparison withE. coliO157 Infections

Shiga toxin-producing Escherichia coli (STEC) are an important cause of diarrhea and the major cause of postdiarrheal hemolytic uremic syndrome. Non-O157 STEC infections are being recognized with greater frequency because of changing laboratory practices. Foodborne Diseases Active Surveillance Network (FoodNet) site staff conducted active, population-based surveillance for laboratory-confirmed STEC infections. We assessed frequency and incidence of STEC infections by serogroup and examined and compared demographic factors, clinical characteristics, and frequency of international travel among patients. During 2000-2010, FoodNet sites reported 2006 cases of non-O157 STEC infection and 5688 cases of O157 STEC infections. The number of reported non-O157 STEC infections increased from an incidence of 0.12 per 100,000 population in 2000 to 0.95 per 100,000 in 2010; while the rate of O157 STEC infections decreased from 2.17 to 0.95 per 100,000. Among non-O157 STEC, six serogroups were most commonly reported: O26 (26%), O103 (22%), O111 (19%), O121 (6%), O45 (5%), and O145 (4%). Non-O157 STEC infections were more common among Hispanics, and infections were less severe than those caused by O157 STEC, but this varied by serogroup. Fewer non-O157 STEC infections were associated with outbreaks (7% versus 20% for O157), while more were associated with international travel (14% versus 3% for O157). Improved understanding of the epidemiologic features of non-O157 STEC infections can inform food safety and other prevention efforts. To detect both O157 and non-O157 STEC infections, clinical laboratories should routinely and simultaneously test all stool specimens submitted for diagnosis of acute community-acquired diarrhea for O157 STEC and for Shiga toxin and ensure that isolates are sent to a public health laboratory for serotyping and subtyping.

Comparison of clinical and epidemiological features of Shiga toxin-producing Escherichia coli O157 and non-O157 infections in British Columbia, 2009 to 2011.

Shiga toxin-producing Escherichia coli (STEC) are major foodborne agents that have the potential to cause severe enteric illnesses and large outbreaks worldwide. Several studies found non-O157 infections to be clinically milder than O157 STEC infections. To compare the clinical and epidemiological profiles of O157 and non-O157 STEC human infections in British Columbia (BC). All STEC cases reported in BC from 2009 to 2011 by four local health authorities were included in the study. Cases were classified according to STEC serotype based on laboratory information. Information was gathered via case interview forms. Data analysis included the χ(2) test and Mann-Whitney test; P<0.05 was considered to be statistically significant. A total of 260 STEC cases were reported, including 154 (59.2%) O157 cases, 63 (24.2%) non-O157 cases and 43 (16.5%) STEC cases with no serotype identified. Hospitalization rate was higher and duration of hospitalization was significantly longer for O157 cases compared with non-O157 cases, but other clinical features were not significantly different. Patients with non-O157 infections were significantly more likely to have travelled outside Canada, less likely to report food exposure at social gatherings and more likely to consume bagged greens and cheese. O157 is the predominant O serotype in BC and appeared to be more clinically severe than non-O157 STEC infections. However, the true incidence and severity of non-O157 remain unknown due to our current inability to detect all non-O157 cases. The present study and the literature suggest the need to identify more predictive virulence factors because serotype does not consistently predict disease severity.

Assessment of Physician Knowledge and Practices Concerning Shiga Toxin–Producing Escherichia coli Infection and Enteric Illness, 2009, Foodborne Diseases Active Surveillance Network (FoodNet)

Shiga toxin-producing Escherichia coli (STEC) infections cause acute diarrheal illness and sometimes life-threatening hemolytic uremic syndrome (HUS). Escherichia coli O157 is the most common STEC, although the number of reported non-O157 STEC infections is growing with the increased availability and use of enzyme immunoassay testing, which detects the presence of Shiga toxin in stool specimens. Prompt and accurate diagnosis of STEC infection facilitates appropriate therapy and may improve patient outcomes. We mailed 2400 surveys to physicians in 8 Foodborne Diseases Active Surveillance Network (FoodNet) sites to assess their knowledge and practices regarding STEC testing, treatment, and reporting, and their interpretation of Shiga toxin test results. Of 1102 completed surveys, 955 were included in this analysis. Most (83%) physicians reported often or always ordering a culture of bloody stool specimens; 49% believed that their laboratory routinely tested for STEC O157, and 30% believed that testing for non-O157 STEC was also included in a routine stool culture. Forty-two percent of physicians were aware that STEC, other than O157, can cause HUS, and 34% correctly interpreted a positive Shiga toxin test result. All STEC knowledge-related factors were strongly associated with correct interpretation of a positive Shiga toxin test result. Identification and management of STEC infection depends on laboratories testing for STEC and physicians ordering and correctly interpreting results of Shiga toxin tests. Although overall knowledge of STEC was low, physicians who had more knowledge were more likely to correctly interpret a Shiga toxin test result. Physician knowledge of STEC may be modifiable through educational interventions.

Outbreak of Shiga Toxin-producing Escherichia coli Serotype O26

Shiga toxin-producing Escherichia coli (STEC) O26:H11 is an emerging cause of disease with serious potential consequences in children. The epidemiology and clinical spectrum of O26:H11 are incompletely understood. We investigated an outbreak of O26:H11 infection among children younger than 48 months of age and employees at a child care center. Every employee at the center (n = 20) and every child <48 months (n = 55) were tested for STEC and administered a questionnaire. Thirty environmental health inspections and site visits were conducted. A cohorting strategy for disease control was implemented. Eighteen confirmed and 27 suspect cases were detected. There were no hospitalizations. The illness rate was 60% for children and employees. The risk of being a case in children <36 months was twice the risk among children of 36 to 47 months (risk ratio: 2.10; 95% confidence interval: 1.00, 4.42). The median duration of shedding among symptomatic confirmed cases was 30.5 days (range: 14-52 days). Four (22%) confirmed cases were asymptomatic and 3 (17%) shed intermittently. Nearly half (49%) of the household contacts of confirmed cases developed a diarrheal illness. The outbreak was propagated by person-to-person transmission; cohorting was an effective disease control strategy. This was the largest reported outbreak of O26:H11 infection in the United States and the largest reported non-O157 STEC outbreak in a US child care center. Non-O157 STEC infection is a differential diagnosis for outbreaks of diarrhea in child care settings. Aggressive disease control measures were effective but should be evaluated for outbreaks in other settings.

Laboratory Practices and Incidence of Non-O157 Shiga toxin–producingEscherichia coliInfections

We surveyed laboratories in Washington State, USA, and found that increased use of Shiga toxin assays correlated with increased reported incidence of non-O157 Shiga toxin–producing Escherichia coli (STEC) infections during 2005–2010. Despite increased assay use, only half of processed stool specimens underwent Shiga toxin testing during 2010, suggesting substantial underdetection of non-O157 STEC infections.

Ten-Year Trends and Risk Factors for Non-O157 Shiga Toxin-Producing Escherichia coli Found Through Shiga Toxin Testing, Connecticut, 2000-2009

The epidemiology over time of non-O157 Shiga toxin-producing Escherichia coli (STEC) is unknown. Since 1999, increasing numbers of laboratories in Connecticut have been testing for ST rather than culturing for O157, enabling identification of non-O157 STEC. Beginning in 2000, Connecticut laboratories were required to submit ST-positive broths to the State Laboratory for isolation and typing of STEC. The ratio of non-O157:O157 from laboratories conducting ST testing was used to determine state-level estimates for non-O157 STEC. Patients with STEC were interviewed for exposure factors in the 7 days preceding illness. Incidence trends, clinical features, and epidemiology of non-O157 and O157 STEC infections were compared. From 1 January 2000 through 31 December 2009, ST testing detected 392 (59%) of 663 reported STEC infections; 229 (58%) of the isolates were non-O157. The estimated incidence of STEC infection decreased by 34%. O157 and the top 4 non-O157 serogroups, O111, O103, O26, and O45, were a stable percentage of all STEC isolates over the 10-year period. Bloody diarrhea, hospitalization, and hemolytic uremic syndrome were more common in patients with O157 STEC than in patients with non-O157 STEC infection. Exposure risks of patients with non-O157 STEC infection differed from those of patients with O157 STEC infection primarily in international travel (15.3% vs 2.5%; P < .01). Non-O157 types differed from each other with respect to several epidemiologic and exposure features. Both O157 and non-O157 STEC infection incidence decreased from 2000 through 2009. Although infection due to O157 is the most common and clinically severe STEC infection, it accounts for a minority of all clinically significant STEC infections. STEC appear to be a diverse group of organisms that have some differences as well as many epidemiologic and exposure features in common.

Public health approach to detection of non-O157 Shiga toxin-producing Escherichia coli: summary of two outbreaks and laboratory procedures.

Routine laboratory testing may not detect non-O157 Shiga toxin-producing Escherichia coli (STEC) reliably. Active clinical, epidemiological, environmental health, and laboratory collaboration probably influence successful detection and study of non-O157 STEC infection. We summarized two outbreak investigations in which such coordinated efforts identified non-O157 STEC disease and led to effective control measures. Outbreak 1 involved illness associated with consuming unpasteurized apple cider from a local orchard. Public health personnel were notified by a local hospital; stool specimens from ill persons contained O111 STEC. Outbreak 2 involved bloody diarrhoea at a correctional facility. Public health personnel were notified by the facility infection control officer; O45 STEC was the implicated agent. These reports highlight the ability of non-O157 STEC to cause outbreaks and demonstrate that a coordinated effort by clinicians, infection-control practitioners, clinical diagnostic laboratorians, and public health personnel can lead to effective identification, investigation, and prevention of non-O157 STEC disease.

Variation in the prevalence of non-O157 Shiga toxin-producing Escherichia coli in four sheep flocks during a 12-month longitudinal study

Shiga toxin-producing Escherichia coli (STEC) are an important group of emerging pathogens, with ruminants recognised as their main natural reservoir. The aim of this longitudinal study was to provide information on the prevalence and existence of seasonal variation in the occurrence of non-O157 STEC in four sheep flocks over a 12-month period. A total of 504 faecal samples from 48 adult sheep in four flocks were collected and examined for STEC using both phenotypic and genotypic methods. STEC were isolated from 407 (80.8%) faecal samples representing all the animals sampled. The overall monthly prevalence of STEC varied between 71.1 and 94.4% and no seasonal variation in the occurrence of STEC could be observed over the study period. A total of 521 STEC isolates were characterised. The PCR procedure indicated that 275 (52.8%) isolates carried the stx 1 gene, 44 (8.4%) carried the stx 2 gene and 202 (38.8%) contained both of these genes. The eae and ehxA genes were detected in 4 (0.8%) and 368 (70.6%) isolates, respectively. The isolates belonged to 28 O serogroups, although 72.4% were restricted to only 10 serogroups (O5, O6, O76, O87, O91, O123, O128, O146, O166 and O176). None of the isolates belonged to the O157 STEC serogroup. STEC isolates of serogroups O33, O53, O105 and O162 have not previously been reported in sheep. This is the first study to report the maintenance of high frequencies of non-O157 STEC infection in sheep flocks over long time periods.

Risk Factors of Sporadic Human Infection of Shiga Toxin Producing Escherichia coli O157 and Non-O157 in Australia

Background: Shiga toxin producing Escherichia coli (STEC) causes acute gastroenteritis characterised by abdominal cramps and bloody diarrhoea. In Australia and elsewhere, serotype O157 is the predominant strain of STEC. To investigate risk factors for STEC O157 and non-O157 infection we conducted an Australia-wide case control study between 2003 and 2007. Methods: Patients infected with STEC that were notified to health departments were eligible for the study and three controls per case were selected from a bank of controls based on sex and five year age groups. Cases were recruited in South Australia from July 2003, with all other Australian states participating from 2005. A standardised questionnaire was used to collect information regarding demographic data, food, behavioural and environmental exposures. All factors associated with O157 and non-O157 infection at the univariate level (p < 0.1) were included in separate logistic regression models. Results: The O157 analysis included 43 cases and 117 controls; the non-O157 analysis included 71 cases and 187 controls. In multivariate analysis, STEC O157 infection was associated with eating hamburgers (OR = 6.6, 95% CI = 2.2–20.0), eating out of the home (OR = 4.8, 95% CI = 1.7–13.6) and living on or visiting a farm (OR = 4.6, 95% CI = 1.2–17.6). Non-O157 STEC infection was associated with consuming chicken deli meat (OR = 5.3, 95% CI = 1.5–18.5), working with animals (OR = 3.2, 95% CI = 1.2-8.3) and asthma (OR = 2.7, 95% CI = 1.1–6.3). Bush camping in Australia had an elevated odds ratio, although this was not significant (OR = 3.3, 95% CI = 0.9–11.9). Conclusions: This national case control study is the first to provide risk factors for both O157 and non-O157 STEC cases. That O157 was associated with farm exposure and eating hamburgers is consistent with that reported internationally. However, some of the risk factors for non-0157 cases have not been reported previously. It is important to consider these risk factors in developing prevention programmes for this serious illness.

Cattle Density and Shiga Toxin-ProducingEscherichia coliInfection in Germany: Increased Risk for Most but Not All Serogroups

Shiga toxin-producing Escherichia coli (STEC) cause severe gastroenteritis and life-threatening hemolytic-uremic syndrome. For STEC of serogroup O157, association between disease incidence and cattle contact has been established in some countries. For other (non-O157) serogroups, however, accounting for approximately 80% of notified STEC gastroenteritis in Germany, the role of cattle in human infection is less clear. For example, an association of non-O157 STEC infection and cattle density has not been investigated. The aim of this study was thus to investigate a potential association between STEC incidence and cattle density in Germany, with special attention to the non-O157 serogroups. We modeled district-level incidence of notified human STEC cases in relation to cattle density, utilizing German notification data from 2001 through 2003. Cattle numbers came from the national "Proof of Origin and Information Database for Animals." A Bayesian Poisson regression model was used, incorporating independent, as well as spatially correlated, district-level random effects into the analysis. We analyzed 3216 German STEC cases. Cattle density was positively associated with overall STEC incidence. The risk for STEC infection increased by 68% per 100 additional cattle/km(2). The magnitude of the risk estimates differed by serogroup and was greatest for O111. A positive association was found for all major disease-causing serogroups (O26, O103, O111, O128, O145, O157) except O91. The association with serogroup O26 (lowest median age of patients) was only borderline significant. Residual variation indicates that additional factors not under study may also be of importance, and that they may be serogroup- and region-specific, too. In conclusion, this study suggests that living in a cattle-raising region appears to imply risk not only for STEC O157, but also for most non-O157 serogroups. Furthermore, the varying magnitude of this risk and the residual variation found for different serogroups indicate that risk profiles for human STEC infection may be serogroup-specific. This needs to be taken into account in risk factor studies for non-O157 STEC, ideally by reporting risks separately by serogroup.

Bacterial Genetic Determinants of Non‐O157 STEC Outbreaks and Hemolytic‐Uremic Syndrome after Infection

Although O157:H7 Shiga toxin-producing Escherichia coli (STEC) are the predominant cause of hemolytic-uremic syndrome (HUS) in the world, non-O157:H7 serotypes are a medically important cause of HUS that are underdetected by current diagnostic approaches. Because Shiga toxin is necessary but not sufficient to cause HUS, identifying the virulence determinants that predict severe disease after non-O157 STEC infection is of paramount importance. Disease caused by O157:H7 STEC has been associated with a 26-gene pathogenicity island known as O island (OI) 122. To assess the public-health significance of this pathogenicity island, we examined the association between OI122 genes and outbreaks and HUS after non-O157 STEC infection. We found that a subset of OI122 genes is independently associated with outbreaks and HUS after infection with non-O157 STEC. The presence of multiple virulence genes in non-O157 serotypes strengthened this association, which suggests that the additive effects of a variable repertoire of virulence genes contribute to disease severity. In vivo, Citrobacter rodentium mutants lacking outbreak- and HUS-associated genes were deficient for virulence in mice; in particular, nleB mutant bacteria were unable to cause mortality in mice. The present study shows that virulence genes associated epidemiologically with outbreaks and HUS after non-O157 STEC infection are pivotal to the initiation, progression, and outcome of in vivo disease.