Application of a real-time PCR-based system for monitoring of O26, O103, O111, O145 and O157 Shiga toxin-producing Escherichia coli in cattle at slaughter

Faecal samples were collected from 573 slaughtered cattle aged between three and 24 months in seven abattoirs. After enrichment (mTSB with novobiocin), samples were screened by real‐time PCR first for stx and if positive, tested for the top‐five Shiga toxin‐producing Escherichia coli (STEC) serogroups using PCR assays targeting genes specific for serogroups O26, O103, O111, O145 and O157. Of 563 samples with available results, 74.1% tested positive for stx genes. Amongst them, the serogroups O145, O103, O26, O157 and O111 were detected in 41.9%, 25.9%, 23.9%, 7.8% and 0.8%, respectively. From 95 O26, 166 O145 and 30 O157 PCR‐positive samples, 17 O26, 28 O145 and 12 O157 strains were isolated by colony hybridization after immunomagnetic separation. The 17 O26 strains were eae‐positive, but only nine strains harboured stx (eight possessing stx1 and one stx2). Of the 28 O145 strains, ten were eae‐positive including four harbouring stx1 or stx2, whereas 18 were negative for stx and eae. Five of the 12 O157 strains harboured stx2 and eae, did not ferment sorbitol, and were identified as STEC O157:H7/H^?. The other seven O157 strains were negative for stx and eae or positive only for eae. Shiga toxin genes and the top‐five STEC serogroups were frequently found in young Swiss cattle at slaughter, but success rates for strain isolation were low and only few strains showed a virulence pattern of human pathogenic STEC.     

A National Surveillance of Shiga Toxin‐Producing Escherichia coli in Food‐Producing Animals in Japan

To assess the public health risk, the prevalence and anti‐microbial resistance of Shiga toxin‐producing Escherichia coli (STEC) among food‐producing animals were studied throughout Japan. Faecal samples were collected from healthy animals of 272 cattle, 179 pigs, and 158 broilers on 596 farms in all 47 Japanese prefectures. STEC were isolated from 62 (23%) cattle and 32 (14%) pig samples but from no chicken samples. Of the bovine isolates, 19 belonged to serotypes frequently implicated in human disease (O157:H7/non‐motile (NM)/H not typeable, O26:NM/H11/H21/H not typeable, O113:H21, and O145:NM). The eae genes were observed in 37% of bovine isolates; among them one O145:NM and all four O157 isolates possessed eae‐γ1, and one O145:NM, one O103:H11, and all five O26 isolates possessed eae‐β1 gene. Among the swine isolates, stx2e were dominant, and serotypes frequently implicated in human diseases or eae‐positive isolates were not observed. Bovine isolates showed less anti‐microbial resistance, but six isolates of 26:NM/H11 and O145:NM were multi‐resistant and may need careful monitoring. Swine isolates showed various resistance patterns; chloramphenicol resistance patterns were more common than in bovine isolates. This first national study of STEC in the Japanese veterinary field should aid our understanding of Japan's STEC status.     

Stochastic Simulation Model Comparing Distributions of STEC O157 Faecal Shedding Prevalence Between Cattle Vaccinated With Type III Secreted Protein Vaccines and Non‐Vaccinated Cattle

Pens of cattle with high Escherichia coli O157:H7 (STEC O157) prevalence at harvest may present a greater risk to food safety than pens of lower prevalence. Vaccination of live cattle against STEC O157 has been proposed as an approach to reduce STEC O157 prevalence in live cattle. Our objective was to create a stochastic simulation model to evaluate the effectiveness of pre‐harvest interventions. We used the model to compare STEC O157 prevalence distributions for summer‐ and winter‐fed cattle to summer‐fed cattle immunized with a type III secreted protein (TTSP) vaccine. Model inputs were an estimate of vaccine efficacy, observed frequency distributions for number of animals within a pen, and pen‐level faecal shedding prevalence for summer and winter. Uncertainty about vaccine efficacy was simulated using a log‐normal distribution (mean = 58%, SE = 0.14). Model outputs were distributions of STEC O157 faecal pen prevalence of summer‐fed cattle unvaccinated and vaccinated, and winter‐fed cattle unvaccinated. The simulation was performed 5000 times. Summer faecal prevalence ranged from 0% to 80% (average = 30%). Thirty‐six per cent of summer‐fed pens had STEC O157 prevalence >40%. Winter faecal prevalence ranged from 0% to 60% (average = 10%). Seven per cent of winter‐fed pens had STEC O157 prevalence >40%. Faecal prevalence for summer‐fed pens vaccinated with a 58% efficacious vaccine product ranged from 0% to 52% (average = 13%). Less than one per cent of vaccinated pens had STEC O157 prevalence >40%. In this simulation, vaccination mitigated the risk of STEC O157 faecal shedding to levels comparable to winter, with the major effects being reduced average shedding prevalence, reduced variability in prevalence distribution, and a reduction in the occurrence of the highest prevalence pens. Food safety decision‐makers may find this modelling approach useful for evaluating the value of pre‐harvest interventions.     

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.     

Influence of processed grains on fecal pH, starch concentration, and shedding of Escherichia coli O157 in feedlot cattle

Manipulation of cattle diets has been proposed as a possible preharvest control measure for Escherichia coli O157. Altering hindgut fermentation through diet changes may be a means to reduce fecal shedding of E. coli O157. In Exp. 1, the objective was to determine whether fecal shedding of E. coli O157 was related to fecal starch concentration. Beginning on d 20, and every week thereafter until d 61, steers in 54 pens (6 to 7 steers per pen) were sampled (n = 122) by fecal collection and rectoanal mucosal swabs (RAMS) for E. coli O157 and fecal starch concentration determinations. Escherichia coli O157 prevalence was 3.3% in fecal samples, 4.1% as measured by RAMS, and 4.9% by fecal or RAMS samples. Steers positive for E. coli O157 contained 21% more (P < 0.05) fecal starch than steers that were negative for E. coli O157. In Exp. 2, we attempted to alter the concentration of starch escaping rumen fermentation by feeding finishing diets based on steam-flaked corn (SFC) and dry-rolled corn (DRC) to 30 heifers prescreened for being culture positive for fecal E. coli O157. Beginning on d 13, heifers were sampled (feces and RAMS) weekly to monitor fecal pH and starch concentration, and prevalence of E. coli O157. Prevalence of E. coli O157 remained above 30% for the first 13 d, but declined (P < 0.05) over the entire 7-wk period. Based on RAMS, the prevalence of E. coli O157 tended to be greater (P = 0.08) for heifers fed SFC than for those fed the DRC diet. After d 20, heifers fed DRC had greater (P < 0.05) fecal starch and lower (P < 0.05) fecal pH than heifers fed SFC. Fecal pH was negatively correlated (r = - 0.34; P < 0.05; n = 143) with fecal starch concentration. Fecal starch concentration and pH were not different (P > 0.05) for heifers that were positive or negative for E. coli O157. Our data suggest that fecal shedding of E. coli O157 was not related to fecal pH or starch concentration in cattle fed grain-based diets.     

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.     

Escherichia coli O157:H7 – Discerning Facts from Fiction: An Integrated Research and Extension Project for Multiple Audiences

The O157:H7 (EcO157) epidemiology of Shiga‐toxin‐producing Escherichia coli (STEC) in cattle is complex, and myths about pre‐harvest control are perpetuated. The objectives of this project were to identify perpetuated misinformation and inform four audiences about evidence‐based risks and pre‐harvest control of EcO157 by addressing: (i) EcO157 epidemiology and pre‐harvest control; (ii) how food safety policy is created; and (iii) how to present accurate information about EcO157. An environmental scan using a daily Internet search helped identify themes for education. A literature review of pre‐harvest control measures contributed to the development of educational materials (fact sheets, website, web presentations and conferences). Conference 1 was a webinar with 315 registrants, 10 countries including 41 US states and four Canadian provinces. Most participants felt confident in using their new knowledge, more than half felt confident enough to answer EcO157 questions from the public and many would recommend the recorded version of the webinar to colleagues. Conference 2 was live in the Washington, DC, area with most participants employed by the US government. All agreed that they better understood pre‐harvest control, how food safety policy was made, and were confident they could create an effective message about STEC pre‐harvest control. Videos were posted and received 348 Internet visitors within 2 months. Conference 3 was a webinar with a live audience and Twitter feeds, targeting people who give nutrition advice. Almost all ranked the programme good to excellent and relevant to their work. About 25% indicated that they would share: ‘grass‐fed beef is not safer than grain‐fed’, 25% would share information on effectiveness of cattle vaccines, and 14% would share information on message mapping. Across all conferences, major changes in knowledge included the following: there is no additional risk of EcO157 shedding from grain‐fed versus grass‐fed cattle, pre‐harvest vaccination is efficacious, and production systems (pasture versus confinement) do not affect EcO157 shedding rates.     

Non‐O157 Shiga Toxin‐producing Escherichia coli Isolated from Diarrhoeic Calves in Argentina

Faecal samples from 76 diarrhoeic calves belonging to 36 farms located in the Pampas plain, Argentina, were examined for Shiga toxin‐producing Escherichia coli (STEC). A total of 15 STEC strains were isolated from 12 (15.8%) calves which came from six different farms. All stx positive strains assayed by PCR were also positives in the Vero cell cytotoxicity test. The majority (60.0%) of the STEC strains carried the stx₁ gene. Twelve (80.0%) of the STEC isolates which belonged to serotypes O5:H‐ (n = 4), O26:H11 (n = 4), O26:H‐ (n = 1), O111:H‐ (n = 2), and O123:H38 (n = 1) were also enterohaemolysin (EHly) positive and carried the gene encoding for intimin (eae). All the stx positive strains were negative for the bfpA gene. Localized adherence to HEp‐2 cells were observed in 83.3% of the eae+ STEC strains. STEC belonging to serotype O5:H‐ showed atypical biochemical properties, including urease production. Urease was also produced by two strains belonging to serotypes O153:H? and non‐typeable, respectively. Resistance to three or more antibiotics was observed in 12 (80.0%) of the STEC isolates. Most of the serotypes of STEC recovered in this survey carried virulence traits that are associated with increased human and bovine pathogenicity. The present study shows that highly virulent STEC strains are being shed by diarrhoeic calves from farms located in a high incidence area of human STEC infections.     

Detection and characterisation of Shiga toxin-producing Escherichia coli other than Escherichia coli O157:H7 in wild ruminants

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 work was to establish the prevalence of non-O157 STEC in free-ranging wild ruminants in the Extremadura region of Spain and to characterise them phenogenotypically. Faecal samples were collected from 243 wild ruminants, including Cervus elaphus, Capreolus capreolus, Dama dama and Ovis musimon and were examined for STEC using both phenotypic (Vero cells) and genotypic (PCR and PFGE) methods.Shiga toxin-producing Escherichia coli were isolated from 58 (23.9%) of the samples and a total of 65 isolates were characterised. A PCR method indicated that 11 (16.9%) strains carried the stx₁ gene, 44 (67.7%) carried the stx₂ gene and 10 (15.4%) carried both these genes. The ehxA gene was detected in 37 (57%) of the isolates but none contained either the eae or saa genes. The isolates were from a total of 12 ‘O’ serogroups, although 80% were restricted to the O2, O8, O128, O146, O166 and O174 serogroups. The most commonly isolated STEC bacteria, which were from the O146 serogroup, exhibited a high degree of polymorphism as indicated by PFGE. Shiga toxin-producing Escherichia coli isolates of serogroups O20, O25, O166, O171, O174 and O176 had not previously been found in wild ruminants. This is the first study to confirm that wild ruminants in Spain are a reservoir of STEC and are thus a potential source of human infection.     

Prevalence and characterization of Shiga toxin-producing Escherichia coli O157 and O26 in beef farms

Rectal content grab samples were collected from 2436 beef cattle reared on 406 beef farms in Japan between November 2007 and March 2008. STEC strains O157 and O26 were isolated from 110 (27.1%) and 7 (1.7%) farms, respectively. Farms that tested positive for STEC O157 were located in 35 out of all 47 Japanese prefectures. This indicates that STEC O157 strains are widespread on beef farms nationwide. Of the 2436 tested beef cattle, 218 (8.9%) and 10 (0.4%) had STEC strains O157 and O26 in the rectal content, respectively. The most common Shiga toxin genes detected in the isolated STEC O157 strains were: stx(2c) alone (32.1%), stx(2)/stx(2c) (27.2%), and stx(1)/stx(2) (21.8%). Almost all of the STEC O157 and STEC O26 strains expressed Shiga toxins (Stx). Most of the STEC O157 and STEC O26 strains possessed eaeA and EHEC-hlyA. These results strongly suggest that STEC strains O157 and O26 from beef cattle would be pathogenic to humans. Therefore, it is important to reduce STEC strains O157 and O26 in beef cattle in order to prevent foodborne disease caused by STEC. The presence of dogs and/or cats on a farm was significantly (P=0.02) associated with the prevalence of STEC O157. More research is needed to clarify the role of dogs and cats.     

Occurrence and Characterization of Escherichia coli O157 Isolated from Cattle in Norway

Faecal samples from 504 imported beef cattle were screened to investigate the occurrence of Escherichia coli O157. The results were compared with those from a previous screening of Norwegian dairy cattle, and the occurrence was found to be higher in the imported beef cattle. The E. coli O157 isolates from the previous and present studies were characterized for the genes encoding for shigatoxin 1 (stx ₁), shigatoxin 2 (stx ₂), the intimin protein (eae) and the flagellar protein H7 (fliC) using PCR analysis, pulsed-field gel electrophoresis (PFGE) with the restriction enzyme XbaI, and bacteriophage lambda RFLP analysis using the PvuII restriction enzyme. The isolates from the dairy and beef cattle could be distinguished by the profiles of the toxin genes and by PFGE patterns. Whether the importation of animals in itself should be regarded as a risk factor for the occurrence of E. coli O157, or whether other management factors contribute to the differences in carrier rates compared to the previous study on domestic cattle, is discussed.     

Cross‐sectional study to estimate the prevalence of enterohaemorrhagic Escherichia coli on hides of market beef cows at harvest

Cattle hides are an important source of enterohaemorrhagic Escherichia coli (EHEC) carcass contamination at slaughter. Seven EHEC serogroups are adulterants in raw, non‐intact beef: EHEC O26, O45, O103, O111, O121, O145 and O157. The objective of this study was to estimate the probability for hide contamination with EHEC among US market beef cows at slaughter and to test the effects of season and geographic region on prevalence of hide contamination. Hides (n = 800) of market cows were swabbed at slaughter immediately after exsanguination, prior to hide removal. Cows were sampled from two geographically distinct beef packing plants during four seasons of 2015. Cattle source was categorized by northern or southern region. Samples were tested for EHEC by a molecular screening assay. The effects of region, season and their interaction on the probability of hide contamination by each EHEC serogroup were tested in separate multilevel multivariable logistic regression models, accounting for the random effect of clustering by plant. Statistical significance was set α = .05. Of 800 total samples, at least one EHEC was detected on 630 (79%) hides. Enterohaemorrhagic E. coli O26 was detected on 129 (16%) of all hides sampled, EHEC O45 on 437 (55%), EHEC O103 on 289 (36%), EHEC O111 on 189 (24%), EHEC O121 on 140 (18%), EHEC O145 on 171 (21%) and EHEC O157 on 89 (11%). Detection of EHEC O26 and EHEC O121 was associated with season. Season and region were associated with detecting EHEC O45 and EHEC O157. Season‐by‐region interactions were associated with the outcome of detecting EHEC O103, EHEC O111 and EHEC O145. Season, region of origin and the interaction of these factors affect hide contamination of market beef cattle at slaughter by EHEC, and each serogroup responds to these factors uniquely.     

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.     

Prevalence and antimicrobial resistance of porcine O157 and non-O157 Shiga toxin-producing Escherichia coli from India

The aims of this study were to determine the prevalence of Shiga toxin-producing Escherichia coli (STEC) strains in pigs as a possible STEC reservoir in India as well as to characterize the STEC strains and to determine the antimicrobial resistance pattern of the strains. A total of 782 E. coli isolates from clinically healthy (n?=?473) and diarrhoeic piglets (309) belonging to major pig-producing states of India were screened by the polymerase chain reaction (PCR) assay for the presence of virulence genes characteristic for STEC, that is, Shiga toxin-producing gene(s) (stx1, stx2), intimin (eae), enterohemolysin (hlyA) and STEC autoagglutinating adhesin (Saa). Overall STEC were detected in 113 (14.4 %) piglets, and the prevalence of E. coli O157 and non-O157 STEC were 4 (0.5 %) and 109 (13.9 %), respectively. None of the O157 STEC isolates carried gene encoding for H7 antigen (fliCh7). The various combinations of virulence genes present in the strains studied were stx1 in 4.6 %, stx1 in combination with stx2 gene in 5.1 %, stx1 in combination with stx2 and ehxA in 0.6 %, stx1 in combination with stx2 and eae in 0.2 % and stx2 alone in 3.7 %. All STEC isolates were found negative for STEC autoagglutinating adhesin (Saa). The number of STEC isolates which showed resistance to antimicrobials such as ampicillin, tetracycline, streptomycin, lincomycin, nalidixic acid, sulfadiazine, penicillin, gentamicin, kanamycin and ceftriaxone were 100, 99, 98, 97, 95, 94, 92, 88, 85 and 85, respectively. Ninety-seven isolates showed resistance to more than 2 antimicrobials, and 8 resistance groups (R1 to R8) were observed. This study demonstrates that pigs in India harbour both O157 and non-O157 STEC, and this may pose serious public health problems in future.     

An investigation on the effect of transport and lairage on the faecal shedding prevalence of Escherichia coli O157 in cattle

The aim of the study was to investigate the effect of transport and lairage on the prevalence of Escherichia coli O157 faecal shedding and the subsequent contamination of beef carcasses. Individual rectal faecal samples were taken from two cohorts of cattle (109 and 59) at the farm before transport and at the abattoir post-transport and lairage. The entire outer and inner surfaces of the carcass of each animal were swabbed immediately following slaughter and dressing. The prevalence of E. coli O157 shedding in cattle sampled at farm, post-transport and lairage was 18% (20), 13% (14) and 12% (13) for cohort A and 1.7% (1), 1.7% (1) and 0 for cohort B, respectively. No E. coli O157 was recovered from the 168 dressed carcasses. In total, 98% (46 of 47) of the E. coli O157 isolates from cohort A were potentially pathogenic to man. Transport and lairage do not cause an increase in the prevalence of E. coli O157 faecal shedding in cattle. This study demonstrates that even positive cohorts of cattle may be slaughtered and processed to produce clean carcasses by following good hygienic practices.     

Prevalence of Shiga-toxigenic Escherichia coli O157:H7 in adult dairy cattle

OBJECTIVE: To describe shiga-toxigenic Escherichia coil O157:H7 (STEC O157:H7) fecal shedding prevalence, seasonal fecal shedding patterns, and site-specific prevalence from the oral cavity, skin, and feces of dairy cattle. DESIGN: Cross-sectional study. ANIMALS: Adult dairy cattle from 13 herds in Louisiana. PROCEDURE: Samples were cultured for STEC O157 by use of sensitive and specific techniques, including selective broth enrichment, immunomagnetic separation, monoclonal antibody-based O:H enzyme immunoassay serotyping, and polymerase chain reaction virulence gene characterization. Point estimates and 95% confidence intervals were calculated for fecal shedding prevalence as well as site-specific prevalence from the oral cavity, skin, and feces. Logistic regression was used to assess seasonal variation and differences at various stages of lactation with respect to fecal shedding of STEC O157 in cattle sampled longitudinally. RESULTS: Summer prevalence in herds in = 13) was 38.5%, with a cow-level prevalence of 6.5%. Among positive herds, prevalence ranged from 3% to 34.6%. Samples from 3 of 5 herds sampled quarterly over 1 year yielded positive results for STEC O157. In herds with STEC O157, an increase in cow-level prevalence was detected during spring (13.3%) and summer (10.5%), compared with values for fall and winter. Site-specific prevalences of STEC O157:H7 from oral cavity, skin, and fecal samples were 0%, 0.7%, and 25.2%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Our data indicated that STEC O157:H7 was commonly isolated from dairy cows in Louisiana, seasonally shed, and isolated from the skin surface but not the oral cavity of cows.     

A randomized controlled trial to evaluate the effects of dietary fibre from distillers grains on enterohemorrhagic Escherichia coli detection from the rectoanal mucosa and hides of feedlot steers

Feeding high levels (≥40% dry matter) of distillers grains may increase the risk for cattle to carry enterohemorrhagic Escherichia coli (EHEC) O157. The mechanism for the increased risk is not known nor whether non‐O157 EHEC are similarly affected. Our objective was to test whether the fibre content or other components of modified distillers grains plus solubles (MDGS) affects the probability for cattle to carry EHEC serogroups of public health importance. A 2 × 2 plus 1 factorial treatment arrangement within a randomized block design was utilized. Within each of four blocks, 25 feedlot pens (n = 8 steers/pen) were assigned randomly to (i) corn‐based control diet; (ii) 20% dry matter (DM) MDGS; (iii) 40% DM MDGS; (iv) corn bran added to corn‐based diet to match fibre of 20% MDGS or (v) 40% MDGS. Rectoanal mucosa swabs (RAMS) were collected on day (d)0, d35, d70 and d105; hide swabs were collected on the last feeding day. Samples were tested for EHEC by a molecular screening assay. The effects of fibre source and fibre level on EHEC carriage were tested using multilevel logistic regression (generalized linear mixed models; α = 0.05). EHEC O45 RAMS detection was associated with fibre level, source and sampling day. EHEC O103 RAMS detection increased by feeding 40% MDGS but not the corresponding corn bran diet. Hide contamination by EHEC O45 or O103 was less likely in cattle fed MDGS compared to corn bran diets. EHEC O111 RAMS detection decreased by feeding 40% MDGS but not by feeding the corresponding corn bran diet. Detection of EHEC O157 or O145 was not associated with dietary factors. Feeding 40% MDGS increased the probability for carriage of some EHEC serogroups but decreased probability of others, which indicated that EHEC serogroups have different risk factors associated with feeding MDGS and little association with dietary fibre.     

Pathogenic Shiga toxin-producing Escherichia coli in the intestine of calves

The purpose of this study was to compare the pathological effects of Shiga toxin-producing Escherichia coli (STEC) that vary in their association with bovine and human disease. Shiga toxin-producing E. coli of serotypes associated with both dysentery in calves and hemolytic uremic syndrome (HUS) in humans (O5:H-, O26:H11, O111:H-, O113:H21) were compared with O157:H7 STEC, which are associated with HUS in humans but not with disease in calves. The STEC were administered orally to 80 day-old chicks and into ligated loops in the ileum and colon of four 2- to 6-day-old calves. Examination of the ceca of the chickens 10 d postchallenge showed no adherence or tissue abnormality for any isolate. The calves were euthanized 8 to 10 h postinoculation, and sections of the intestinal loops were examined by light microscopy, transmission and scanning electron microscopy, and immunohistochemistry. All strains showed consistent focal adherence associated with mild lesions in the colon. Attaching and effacing lesions were observed with the eae-positive strains. Ileal lesions were similar to the colonic ones but were sometimes severe, with marked polymorphonuclear leukocyte proliferation in the lamina propria. It is concluded that chickens were unsuitable for studying interaction of STEC with the intestine and that there was no difference in the interaction of the ligated calf intestine with STEC of serotypes associated with disease in calves compared with O157:H7 STEC.     

Isolation of shiga-toxigenic Escherichia coli O157 from hide surfaces and the oral cavity of finished beef feedlot cattle

OBJECTIVE: To determine whether viable shiga-toxigenic Escherichia coli (STEC) O157 could be isolated from hide surface locations and the oral cavity of finished beef feedlot cattle. DESIGN: Within-animal prevalence distribution survey. ANIMALS: 139 finished cattle in 4 pens in a feedlot in Nebraska; prevalence of fecal STEC O157 shedding ranged from 20 to > 90%. PROCEDURE: Samples were collected from 7 sites from each animal: feces, oral cavity, and 5 hide surface locations (lumbar region, ventral aspect of the neck, ventral abdominal midline [ventrum], dorsal thoracic midline [back], and distal aspect of the left hind limb [hock]). RESULTS: Viable STEC O157 were isolated from the oral cavity or 1 or more hide surfaces of 130 cattle, including 50 fecal isolation-negative cattle. Site-specific prevalence of STEC O157 was 74.8% for oral cavity samples, 73.4% for back samples, 62.6% for neck samples, 60.4% for fecal samples, 54.0% for flank samples, 51.1% for ventrum samples, and 41.0% for hock samples. Only 5 cattle tested negative for STEC O157 at all 7 sites. Multiple correspondence and cluster analyses demonstrated that bacterial culture of feces, oral cavity samples, and back samples detected most cattle with STEC O157. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that viable STEC O157 may be isolated from the oral cavity, multiple hide surfaces, and feces of a high percentage of fed beef cattle and that bacterial culture of feces alone generally underestimates the percentage of fed beef cattle from which STEC O157 can be isolated.     

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.