Significance of Escherichia coli O157 in sheep at slaughter in Switzerland

The importance of latent zoonoses has increased in recent years in view of foodborne diseases: (i) the "healthy" animal repesents a reservoir for specific pathogens; () no pathological-anatomical changes in the carcass and its organs show the presence of these pathogens; and (iii) these pathogens may enter the food chain via hygienic weak points in the slaughtering process. To estimate the risks involved and to take appropriate measures, analysis of the slaughtering process should be complemented by collecting data relating to the carriage of the animals of latent zoonotic pathogens. From October 2004 to June 2005, fecal samples from 630 slaughtered sheep were enriched and then examied by IMS technique and by PCR to assess the prevalence of E. coli O157 (OE). Seven samples (1.1%), distributed throughout the whole examination period, were found to be positive. To assess the potential pathogenicity for humans, E. coli O157 strains were isolated by colony hybridization and further characterized. The isolated strains fermented Sorbitol, showed four different H tys (H7, H12, H38, H48), and were all negative for stx. One O157:H7 strain harbored the gene for intimin (eae) in combination with ehxA, and paa. In consequence, the potential health hazard from sheep meat related to O157 STEC seems current not to be of particular importance in Switzerland. Results emphasize the fact that E. coli O157 are not always STEC but may belong to other pathotypes as nontraditional EPEC.     

Risk factors for hide contamination of Scottish cattle at slaughter with Escherichia coli O157

In the slaughter processing of cattle, contaminated hides have been identified as one of the major sources of Escherichia coli O157 carcase contamination. Logistic regression analysis was applied to data collected in a large scale study in Scotland involving 222 cattle forming 34 groups sent for slaughter from 30 farms to 10 slaughterhouses. Aspects of individual animal characteristics, farm management practices and slaughterhouse features were examined to identify potential risk factors for hide contamination at harvest. Two models were developed, the first in which slaughterhouse was modelled as a fixed effect, and a second model where slaughterhouse and farm groups were modelled as random effects. In the first model, there was a significantly increased risk of a carcase testing positive for E. coli O157 on the hide if either the hide of the carcase immediately before or after it on the line was contaminated (OR 3.6; 95% CI: 1.4–9.9). If both adjacent carcases had contaminated hides, the odds ratio for the study carcase having a contaminated hide rose to 11.5 (95% CI: 4.4–32.5). If animals were held in lairage, receiving hay as feed appeared to have a protective effect on hide contamination. Transportation to the slaughterhouse by haulier, as opposed to transport by the farmer, was associated with a 5.4 increase in the odds of E. coli O157 contamination. The use of a crush in the lairage, often employed when reading ear tags, was also found to significantly increase the odds of hide contamination with E. coli O157. In the second model, the inclusion of slaughterhouse and farm group as random effects resulted in two of the previously identified factors being associated with hide contamination. If at least one of the adjacent carcases on the line had a contaminated hide, the associated odds ratio was 6.6 (95% CI: 2.8–15.9), which rose to 22.7 (95% CI: 9.3–55.5) if both adjacent hides were contaminated. Receiving hay in lairage was found to be important to the model, although not significant in itself (OR 0.005; 95% CI: 1.2e⁻⁶–20.7). These results suggest that modifiable risk factors for hide contamination exist. However, in order best to reduce the prevalence of hide contamination at slaughter, individual slaughterhouse risk assessment and intervention strategies are appropriate.     

Escherichia coli in the rumen and colon of slaughter cattle,with particular reference to E. coli O157

The distribution of Escherichia coli O157 and of total E. coli was surveyed in the digestive tract of cattle under 30 months of age, slaughtered between August 1999 and May 2000 in three abattoirs in southern England. Samples were taken from the dorsal and ventral rumen wall, the rumen contents, the colon wall and colon contents, and from faeces or caudal rectal contents. Gut wall samples were processed by vortex-mixer to release loosely adherent bacteria, and by Stomacher to release firmly attached bacteria. E. coli O157 was detected by immunomagnetic separation followed by growth on selective culture media. The numbers of E. coli were higher in the colon than the rumen, and most were located in the digesta phase, rather than associated with the gut wall. The number of E. coli found in the gut and in faeces decreased during the winter months. E. coli O157 was detected more frequently in the colon than in the rumen, but the majority of detections(7/8) were in samples of rumen wall.     

Environmental sources and transmission of Escherichia coli O157 in feedlot cattle

A study was conducted in 2 feedlots in southern Alberta to identify environmental sources and management factors associated with the prevalence and transmission of Escherichia coli O157:H7. Escherichia coli O157:H7 was isolated in preslaughter pens of cattle from feces (0.8%), feedbunks (1.7%), water troughs (12%), and incoming water supplies (4.5%), but not from fresh total mixed rations. Fresh total mixed rations did not support the growth of E. coli O157:H7 and E. coli from bovine feces following experimental inoculation. Within a feedlot, the feces, water troughs, and feedbunks shared a few indistinguishable subtypes of E. coli O157:H7. A few subtypes were repeatedly isolated in the same feedlot, and the 2 feedlots shared a few indistinguishable subtypes. The prevalence of E. coli O157:H7 in water troughs of preslaughter cattle in 1 feedlot was associated with season, maximum climatic temperatures the week before sampling; total precipitation the week before sampling, and coliform and E. coli counts in the water trough.     

Escherichia coli O157:H7 and non-O157 Shiga toxin-producing E. coli in healthy cattle, sheep and swine herds in Northern Spain

Three-hundred and forty-five herds (17 swine, 122 dairy sheep, 124 beef and 82 dairy cattle) were investigated for prevalence of Shiga toxin-producing Escherichia coli (STEC). Rectal faecal samples were selectively enriched and then examined by immunodetection techniques (Immunomagnetic Separation with anti-E. coli O157 Dynabeads, ImmunoMagnetic cell Separation (IMS) and automated enzyme-linked fluorescent immunoassay using VIDAS) and polymerase chain reaction (PCR) (rfbE and fliC genes) to assess the prevalence of E. coli O157:H7. Prevalence of non-O157 STEC was estimated by PCR screening for stx genes of 10 lactose-positive colonies grown on MacConkey agar after enrichment. PCR was used on all STEC isolates to detect stx(1), stx(2), eaeA and E-hlyA genes. Both immunodetection methods showed a moderate-good level of agreement (kappa = 0.649) but IMS showed 87.5% complementary sensitivity. Prevalence of positive herds for E. coli O157:H7 was estimated at 8.7% for sheep and 3.8% for cattle, whereas all the porcine herds tested negative. Non-O157 STEC were also absent from swine, but were isolated more frequently from ovine (50.8%) than bovine herds (35.9%). Within-herd prevalences of excretion of E. coli O157:H7 established by individual testing of 279 sheep (six herds) and 30 beef cattle (one herd) were 7.3% and 6.7% respectively. PCR analysis of 49 E. coli O157:H7 and 209 non-O157 isolates showed a different distribution of virulence genes. All E. coli O157:H7 were stx(2) gene-positive, eaeA was detected in 95.9%, and the toxigenic profile stx(2)/eaeA/E-hlyA was present in 75.5% of the isolates. Among the non-O157 STEC, prevalence of eaeA was significantly lower (5.3%) and E-hlyA was present in 50.2% of the isolates but only sporadically associated with eaeA. stx(2) was predominant in non-O157 isolates from cattle, whereas in sheep the combination stx(1)/stx(2) was more prevalent. This study demonstrated the wide distribution of STEC in ruminant herds, which represent an important reservoir for strains that pose a potential risk for human infections.     

Risk factors associated with faecal shedding of verocytotoxin-producing Escherichia coli O157 in eight known-infected Danish dairy herds

A risk-factor study was performed in eight dairy herds found to excrete verocytotoxin-producing Escherichia coli (VTEC) O157 in a former prevalence study. Associations between excretion of VTEC O157 and management factors such as housing and feeding were analysed in a generalised linear mixed model. The animals were stratified in three age groups and sampled four times during 1 year. The risk of excreting VTEC O157 was higher among weaned calves than non-weaned calves. Among the calves aged 1–4 months, the risk was reduced if the calf had suckled colostrum from the mother or if the calf had stayed >2 days with the mother after calving. Calves aged 5–24 months that had been moved within the last 2 weeks had a higher risk, but risk was reduced if fed barley silage. Cows fed grain or molasses had a higher risk of excreting VTEC O157.     

Prevalence of faecal excretion of verocytotoxigenic Escherichia coli O157 in cattle in England and Wales

During the decade to 1999, the incidence of human infections with the zoonotic pathogen verocytotoxin-producing Escherichia coli O157 (VTEC O157) increased in England and Wales. This paper describes the results of a survey of 75 farms to determine the prevalence of faecal excretion of VTEC O157 by cattle, its primary reservoir host, in England and Wales. Faecal samples were collected from 4663 cattle between June and December 1999. The prevalence of excretion by individual cattle was 4.2 per cent (95 per cent confidence interval [CI] 2.0 to 6.4) and 10.3 per cent (95 per cent CI 5.8 to 14.8) among animals in infected herds. The within-herd prevalence on positive farms ranged from 1.1 to 51.4 per cent. At least one positive animal was identified on 29 (38.7 per cent; 95 per cent CI 28.1 to 50.4) of the farms, including dairy, suckler and fattening herds. The prevalence of excretion was least in the calves under two months of age, peaked in the calves aged between two and six months and declined thereafter. The phage types identified most widely were 4, 34 and 2, which were each found on six of the 29 positive farms.     

Transmission of Escherichia coli O157:H7 to cattle by house flies

The main reservoir of Escherichia coli O157:H7 is the digestive tract of cattle; however, the ecology of this food-borne pathogen is poorly understood. House flies (Musca domestica L.) might play a role in dissemination of this pathogen in the cattle environment. In our study, eight calves were individually exposed to house flies that were orally inoculated with a mixture of four strains of nalidixic acid-resistant E. coli O157:H7 (Nal(R)EcO157) for 48h. Another eight calves were individually exposed to uninoculated flies and served as the control. Fresh cattle feces (rectal sampling) and drinking water were periodically sampled and screened for Nal(R)EcO157 up to 19 days after the exposure. At the end of the experiment, all calves were euthanized and the lumen contents of rumen, cecum, colon, and rectum as well as swab samples of gall-bladder mucosa and the recto-anal mucosa were screened for Nal(R)EcO157. On day 1 after the exposure, fecal samples of all eight calves and drinking-water samples of five of eight calves exposed to inoculated flies tested positive for Nal(R)EcO157. The concentration of Nal(R)EcO157 in feces ranged over time from detectable only by enrichment (<10(2)) to up to 1.1 x 10(6)CFU/g. Feces of all calves remained positive for Nal(R)EcO157 up to 11 days after the exposure and 62% were positive until the end of experiment. Contamination of drinking water was more variable and all samples were negative on day 19. At necropsy, the highest prevalence of Nal(R)EcO157 was in the recto-anal mucosa region, followed by rectal and colonic contents.     

Pre-slaughter control of Escherichia coli O157 in beef cattle: a simulation study.

A stochastic simulation model was used to assess the benefit of measures implemented in the pre-slaughter period that are aimed at reducing the contamination of beef carcasses with Shiga-like-toxin-producing Escherichia coli O157. The scenario studied was based on an abattoir processing approximately 1000 head of lot-fed cattle per day. Input assumptions were described using probability distributions to reflect uncertainty in their true values. Control measures that were assessed were based on either a reduction in herd prevalence of infection, reduction in opportunity for cross-contamination in the processing plant by re-ordering of the slaughter queue, reduction of concentration of E. coli O157 in fresh faeces, or a reduction in the amount of faeces, mud and bedding ('tag') transferred from the hide to the carcass. Some control measures evaluated were hypothetical in nature and were included to assist with the planning of research priorities. Simulations suggested that the greatest potential impact is associated with vaccination and with an agent that reduces shedding E. coli O157 in faeces. Knowledge of herd-test results obtained by testing a sample of animals from the herd provides only a minor advantage in control programmes, although application of a rapid test to all animals in all lots might be of some benefit. Under most scenarios, there is ample opportunity for cross-contamination to occur within the slaughter plant as a result of early entry of cattle contaminated with E. coli O157. An industry-wide reduction in the amount of tag attached to hides and addition of a source of cattle having a prolonged average fasting time were not predicted to have a large impact on mean amount of carcass contamination with E. coli O157.     

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。     

Escherichia coli O157 in feedlot cattle feces and water in four major feeder-cattle states in the USA

The prevalence of Escherichia coli O157 was determined in 10662 fecal samples, 2130 water and 1132 water tank-sediment samples collected during the summer months in 2001 from 711 pens in 73 feedlots located in Kansas, Nebraska, Texas, or Oklahoma, USA. Overall, 10.2% of fecal samples were positive for E. coli O157, with 52% of the pens and 95.9% of the feedlots having at least one positive fecal sample. There were no differences among states or months in the fecal prevalences. Water or water tank-sediment was positive in 13.1% of the water tanks, and 60.3% of feedlots had at least one positive tank. Cattle were more likely to be shedding E. coli O157 in pens with positive water tanks, and water was more likely to be positive when E. coli O157 was detected in the sediment.     

Distribution of Escherichia coli O157:H7 within and among cattle operations in pasture-based agricultural areas

OBJECTIVE: To determine the distribution of Escherichia coli O157:H7 in pasture-based cattle production areas. SAMPLE POPULATION: Two 100-km2 agricultural areas consisting of 207 pasture, 14 beef-confinement, and 3 dairy locations within 24 cattle operations. PROCEDURE: 13,726 samples from cattle, wildlife, and water sources were obtained during an 11-month period. Escherichia coli O157:H7 was identified by use of culture and polymerase chain reaction assays and characterized by pulsed-field gel electrophoresis (PFGE). RESULTS: Odds of recovering E coli O157:H7 from feeder-aged cattle were > 4 times the odds for cow-calf or dairy cattle. There was no difference in prevalence for pastured versus confined cattle after controlling for production age group. Number of samples collected (37 to 4,829), samples that yielded E coli O157:H7 (0 to 53), and PFGE subtypes (0 to 48) for each operation varied and were highly correlated. Although most PFGE subtypes were only detected once, 17 subtypes were detected on more than 1 operation. Ten of 12 operations at which E coli O157:H7 was detected had at least 1 subtype that also was detected on another operation. We did not detect differences in the probability of having the same subtype for adjacent operations, nonadjacent operations in the same study area, or operations in the other study area. CONCLUSIONS AND CLINICAL RELEVANCE: Strategies aimed at controlling E coli O157:H7 and specific subtypes should account for the widespread distribution and higher prevalence in feeder-aged cattle regardless of production environment and the fact that adjacent and distant cattle operations can have similar subtypes.     

Evaluation of a model for Escherichia coli O157:H7 colonization in streptomycin-treated adult cattle

OBJECTIVE: To develop a repeatable model for studying colonization with streptomycin-resistant Escherichia coli O157:H7 in adult cattle. ANIMALS: 5 adult mixed-breed beef cattle. PROCEDURES: Cattle were surgically cannulated in the duodenum, treated daily with streptomycin (33 mg/kg) via the duodenal cannula prior to and during experimental colonizations, and colonized with 10(10) CFUs of streptomycin-resistant E coli O157:H7 via the duodenal cannula. Colonization of rectal mucus and shedding in feces were monitored. Antimicrobials were administered to eliminate the colonizing strain so that 5 repeated colonization experiments could be performed. A comprehensive analysis of colonization was performed at necropsy. RESULTS: Streptomycin treatment resulted in improved experimental colonization variables, compared with untreated controls, during initiation (days 2 to 6) and early maintenance (days 7 to 12) of colonization. Elimination of the colonizing strain followed by 5 repeated colonizations in the same animals indicated the repeatability of the protocol. Positive results of bacteriologic culture of feces 7 and 12 days after colonization were obtained in 100% and 84% of samples, respectively, across all animals and trials. At necropsy, highest magnitude recovery was in terminal rectal mucus. CONCLUSIONS AND CLINICAL RELEVANCE: The model was highly repeatable and novel with respect to streptomycin treatment, use of duodenal cannulas, and repeated colonizations of the same animals. Its use in adult cattle, from which most bovine-derived food originates, is critical to the study of preharvest food safety. The findings have implications for understanding intermittency of shedding in the field and for proposed vaccine-based interventions.