A statewide outbreak of Escherichia coli O157:H7 infections in Washington State

In November 1986, a statewide outbreak of Escherichia coli O157:H7 infections in Washington State was identified after a physician in an eastern Washington community hospitalized three patients with hemorrhagic colitis which progressed to thrombotic thrombocytopenic purpura. Epidemiologic investigation identified 37 cases in this community and linked the illnesses to a local restaurant which had served ground beef that was the suspected initial vehicle of transmission. The plasmid profile and toxin production pattern (Shiga-like toxin II alone) of the outbreak strain provided a unique strain marker. E. coli O157:H7 infections caused by this strain were simultaneously seen in other parts of the state among nursing home residents and in patients with the hemolytic-uremic syndrome, and an increase in sporadic cases of hemorrhagic colitis was noted at a Seattle health maintenance organization. It is suspected that a contaminated product, probably ground beef distributed statewide, was the common source. Tracing of this meat led to farms where rectal swabs from six (1%) of 539 cattle tested yielded E. coli O157:H7, although the plasmids and toxin production patterns of these isolates differed from the human outbreak strain. Introduction of a single strain of E. coli O157:H7 has the potential to cause widespread concurrent outbreaks. Such outbreaks are likely to escape recognition until heightened screening and surveillance for E. coli O157:H7 is established.     

Escherichia coli O157:H7 outbreak associated with consumption of ground beef, June-July 2002

OBJECTIVE: A case-control and environmental study tested the hypothesis that purchasing and eating ground beef from a specific source was the cause of a cluster of cases of hemolytic uremic syndrome (HUS) and Escherichia coli (E. coli) O157:H7 gastroenteritis. METHODS: A case-control study comparing risk factors was conducted over the telephone on nine case-patients with 23 selected controls. An environmental investigation was conducted that consisted of reviewing beef handling practices at a specific local supermarket and obtaining ground beef samples from the store and two households with case-patients. RESULTS: The analysis of the case-control study showed that eight case-patients (89%) purchased ground beef at Grocery Chain A compared with four controls who did not develop illness (17%) (matched odds ratio=undefined; 95% confidence interval 2.8, infinity; p=0.006). The environmental investigation showed that Grocery Chain A received meat from Meatpacker A. Laboratory analysis of meat samples from Meatpacker A and Grocery Chain A and stool samples from some patients recovered an identical strain of E. coli O157:H7 according to pulse-field gel electrophoresis. CONCLUSIONS: Both the case-control and environmental studies showed that purchasing ground beef at Grocery Chain A, which received ground beef from Meatpacker A, was the major risk factor for illness in eight case-patients; the ninth case-patient was found to be unrelated to the outbreak. Furthermore, meat from Meatpacker A was associated with a nationwide outbreak of E. coli O157:H7 illness that resulted in the second largest recall of beef in U.S. history at the time.     

Detection of Escherichia coli O157:H7 and Listeria monocytogenes in beef products by real-time polymerase chain reaction

Rapid methods for the detection of Escherichia coli O157:H7 and Listeria monocytogenes in food products are important to the food industry and for public health. Conventional microbiological methods and newly developed molecular-based techniques such as polymerase chain reaction (PCR)-based methods are time consuming. In this study, a faster method based on utilization of a hybridization probe with real-time PCR, was developed and applied for detection of E. coli O157:H7 and L. monocytogenes from artificially contaminated raw ground beef and fully cooked beef hotdogs. Target genes for E. coli O157:H7 and L. monocytogenes were rfbE and hylA, respectively. An analysis of 169 bacterial strains showed that the chosen primers and probes were specific for detection of E. coli O157:H7 and L. monocytogenes by real-time PCR. The assay was positive for nine of 10. E. coli O157:H7 strains, and all L. monocytogenes (7/7) strains evaluated. Bacterial strains lacking these genes were not detected by these assays. Detection limits of real-time PCR assays ranged from 10(3) to 10(8) colony forming units (CFU)/ml for E. coli O157:H7 in modified tryptic soy broth and 10(4) to 10(8) CFU/mL for L. monocytogenes in Fraser Broth. Detection sensitivity ranged from 10(3) to 10(4) CFU/g of raw ground beef or hotdog without enrichment for E. coli and L. monocytogenes. Approximately 1.4-2.2 CFU/g of E. coli O157:H7 in raw ground beef were detected following an enrichment step of 4 h. Approximately 1.2-6.0 CFU/g of L. monocytogenes in beef hotdogs were detected following an enrichment step of 30 h. The real-time PCR assays for detection of E. coli O157:H7 and L. monocytogenes in raw ground beef and beef hotdogs were specific, sensitive and rapid.     

Polymerase chain reaction screening for Salmonella and enterohemorrhagic Escherichia coli on beef products in processing establishments

Pathogenic Escherichia coli strains on raw or insufficiently cooked foods are of public health concern as serious disease may result from their ingestion. Therefore, many commercial producers of beef products screen for E. coli O157:H7 before shipment. While Salmonella is not considered an adulterant on raw beef products, it is used as an indication of process control. To detect these microorganisms, rapid screening methods are often used to provide results within 8-24 hours after sampling. During 2005-2008, about 971,389 samples from several commercial beef production plants were tested using a rapid screening method based on the polymerase chain reaction to determine if they were presumptively positive for bacterial cells carrying Salmonella or Shiga toxin-producing E. coli-specific genes. Of the product lots sampled (trim, ground beef, and variety meats), 15% were positive for the stx(1) and/or stx(2) (Shiga toxin genes), 9.1% for the eae gene (the attaching and effacing gene [eae] encoding intimin), 3.0% for an rfb gene region (encoding the O157-specific O side chain polysaccharide), and 1.67% for Salmonella by the polymerase chain reaction assay. In general, lots of ground beef showed the lowest frequency of contamination, and variety meats (by-products of carcass evisceration), the highest. Overall, 4.6%, 4.6%, and 0.8% samples were screen-positive for enteropathogenic E. coli, enterohemorrhagic E. coli, and E. coli O157, respectively. Of the E. coli O157-positive samples, 14% were also Salmonella positive. The frequency of screen-positive samples increases during the summer months, probably because of the prevalence of climatic conditions more conducive to microbial growth. The presence of fecal organisms in beef products suggests a failure of sanitary controls during processing and the more prevalent relatives of E. coli O157, Shiga toxin-producing Escherichia coli, enteropathogenic E. coli, and enterohemorrhagic E. coli, serve as more sensitive indicators of contamination than O157 strains alone.     

Cattle as a possible source of verocytotoxin-producing Escherichia coli O157 infections in man.

In May-June 1992 cases of infection with verocytotoxin-producing (VT+) Escherichia coli O157 in South Yorkshire could have been associated with prior consumption of beef from a local abattoir. During investigation of the abattoir, bovine rectal swabs and samples of meat and surface swabs from beef carcasses were examined for E. coli O157, isolates of which were tested for toxigenicity, plasmid content and phage type. E. coli O157 was isolated from 84 (4%) of 2103 bovine rectal swabs; of these 84, 78 (93%) were VT+, the most common phage types being 2 and 8, the types implicated in the cluster of human cases. Positive cattle were from diverse sources within England. E. coli O157 was isolated from 7 (30%) of 23 carcasses of rectal swab-positive cattle and from 2 (8%) of 25 carcasses of rectal swab-negative cattle. The study has shown that cattle may be a reservoir of VT+ E. coli O157, and that contamination of carcasses during slaughter and processing may be how beef and beef products become contaminated and thereby transmit the organism to man.     

Hamburger-associated Escherichia coli O157:H7 infection in Las Vegas: a hidden epidemic.

OBJECTIVES: This study sought to determine whether a multistate fast food hamburger-associated outbreak of Escherichia coli O157:H7 infection involved Las Vegas residents as well and, if so, why public health officials had not detected it. METHODS: A matched case-control study was conducted among persons with bloody diarrhea and their healthy meal companions. Hamburger production, distribution, and cooking methods were reviewed. Unused hamburger patties were cultured, and E. coli O157:H7 isolates were characterized. Local laboratory stool culture practices were reviewed. RESULTS: Fifty-eight cases of bloody diarrhea were identified. Illness was associated with eating regular hamburgers (matched odds ratio [OR] = 9.0, 95% confidence interval [CI] = 1.02,433.4), but 25% of ill persons reported eating only jumbo hamburgers. Regular and jumbo hamburger patties yielded E. coli O157:H7 indistinguishable from the lone clinical isolate. No local laboratory cultured routinely for E. coli O157:H7 until after the outbreak. CONCLUSIONS: A large outbreak of E. coli O157:H7 infections escaped timely notice in Las Vegas because local laboratories did not culture for this pathogen. Health officials should encourage laboratories to screen at least all bloody stools on sorbitol-MacConkey medium.     

Effects of gamma ray and electron-beam irradiations on survival of anaerobic and facultatively anaerobic bacteria

An extension of the approval for food irradiation is desired due to the increase in the incidence of food poisoning in the world. One anaerobic (Clostridium perfringens) and four facultatively anaerobic (Bacillus cereus, Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella Enteritidis) bacteria irradiated with gamma ray or electron beam (E-beam) were tested in terms of survival on agar under packaging atmosphere. Using pouch pack, effects of two irradiations on survival of anaerobic and facultatively anaerobic bacteria were evaluated comparatively. E-beam irradiation was more effective than gamma ray irradiation in decreasing the D10 value of B. cereus at 4 degrees C, slightly more effective in that of E. coli O157, and similarly effective in that of the other three bacteria at 4 degrees C. The gamma irradiation of the bacteria without incubation at 4 degrees C before irradiation was more effective than that of the bacteria with incubation overnight at 4 degrees C before irradiation in decreasing the D10 values of these bacteria (B. cereus, E. coli O157, and L. monocytogenes). Furthermore, ground beef patties inoculated with bacteria were irradiated with 1 kGy by E-beam (5 MeV) at 4 degrees C. The inoculated bacteria in the 1-9 mm beef patties were killed by 1 kGy E-beam irradiation and some bacteria in more than 9 mm beef patties were not killed by the irradiation.     

Comparison between human and animal isolates of Shiga toxin-producing Escherichia coli O157 from Australia

There is very little human disease associated with enterohaemorrhagic Escherichia coli O157 in Australia even though these organisms are present in the animal population. A group of Australian isolates of E. coli O157:H7 and O157:H- from human and animal sources were tested for the presence of virulence markers and compared by XbaI DNA macrorestriction analysis using pulsed-field gel electrophoresis (PFGE). Each of 102 isolates tested contained the gene eae which encodes the E. coli attaching and effacing factor and all but one carried the enterohaemolysin gene, ehxA, found on the EHEC plasmid. The most common Shiga toxin gene carried was stx2c, either alone (16%) or in combination with stx1 (74%) or stx2 (3%). PFGE grouped the isolates based on H serotype and some clusters were source specific. Australian E. coli O157:H7 and H- isolates from human, animal and meat sources carry all the virulence markers associated with EHEC disease in humans therefore other factors must be responsible for the low rates of human infection in Australia.     

2001年中国食源性致病菌及其耐药性主动监测研究

目的　旨在监测中国食源性致病菌及其耐药性。方法　中国疾病预防控制中心营养与食品安全所建立的全国食品污染物监测网的食源性致病菌监测部分 ,2 0 0 1年在全国 11个省市设点采样并检测了七大类 (生肉、熟肉、生奶、冰激淋、酸奶、水产品和蔬菜 )共 4 0 34份样品。结果　沙门氏菌、大肠杆菌O15 7∶H7和单核细胞增生李斯特氏菌 ,总阳性率 5 5 0 %。其中沙门氏菌 3 32 % ,单核细胞增生李斯特氏菌 1 2 9% ,大肠杆菌O15 7∶H70 82 %。生肉的污染情况最为严重 ,阳性率 12 96 %。共分离出沙门氏菌 137株 ,德比沙门氏菌、阿贡纳沙门氏菌、肠炎沙门氏菌、里定沙门氏菌、鸭沙门氏菌、明斯特沙门氏菌及鼠伤寒沙门氏菌 7种血清型占 80 %。结论　沙门氏菌的血清型分布和耐药性各个省不同。自生、熟肉中分离到O15 7∶H7的强毒株。对分离的沙门氏菌、大肠杆菌检测了 14种抗生素的耐药性 ,结果表明沙门氏菌和大肠杆菌中都有多重耐药株。     

大肠杆菌O157多克隆抗体及食品中双抗ELISA测定方法的研究

本研究获得了抗肠出血性大肠杆菌O157:H7的多克隆抗体 ,建立了一种适宜食品样品检测的双抗ELISA检测方法。该方法对纯培养菌液检出限为 10 3 ～ 10 4 cfu ml;只对O157菌株有特异性反应 ,对非O157菌株无交叉反应 ;经过增菌 ,鸡肉与牛奶染菌样品中的大肠杆菌O157的检出限均为 0 1cfu g(cfu ml)。     

绿色荧光蛋白在大肠杆菌O157∶H7检测中的应用

目的　将绿色荧光特征引入大肠杆菌E coliO15 7∶H7,用于传统检测方法的改进及目的菌的应用研究。方法　将pGFP质粒转化E coliO15 7∶H7,构建一种工程菌 (E coliO15 7∶H7 pGFP) ,并进行了选择性培养基的筛选。以E coliO15 7∶H7 pGFP对鸡肉与牛奶等食品样品进行接种与回收试验 ,同时设定不同温度模拟食品储存的不同情况。结果　pGFP质粒可以在E coliO15 7∶H7菌株中稳定存在。将E coliO15 7∶H7 pGFP接种食品样品 ,以LBan培养基平板回收发现 :较高温度存放肉类与牛奶 ,其污染的E coliO15 7∶H7菌量可在 12h增加 35 0 0 0～ 2 0 0 0 0 0倍 ,而 4℃存放可以使污染菌量缓慢下降。结论　构建了一种具有紫外灯下发出绿色荧光与氨苄青霉素抗性等特性的重组菌株———O15 7∶H7 pGFP ,并设计了适合E coliO15 7∶H7 pGFP生长的选择性培养基———LBan。该重组菌株可应用于检测方法的改进及该菌特性的研究 ,是一种非常有用的工程菌     

食品标本处理方法对大肠埃希菌检出限影响

目的 研究快速、敏感、低成本的食品标本处理方法。方法 在牛肉馅中接种不同浓度的大肠埃希菌O157：H7，经离心、过滤，去除PCR抑制剂。浓缩菌株，用煮沸法和酶／冻融法裂解菌株，制备模板DNA。通过PCR检测大肠埃希菌O157：H7志贺毒素基因以评价该处理方法的可行性。结果 在未增菌条件下，PCR最低能检测到103CFU／g牛肉馅，增菌6h，最低能检测1CFU／g牛肉馅，整个检测过程只需要12h。结论 所采用的标本处理方法是一种灵敏、省时、低成本的方法，能显著地提高PCR检测食品标本中的大肠埃希菌O157：H7的灵敏度。     

Review of epidemiological surveys on the prevalence of contamination of healthy cattle with Escherichia coli serogroup O157:H7.

This paper gathers and critically analyses the results of 26 published epidemiological surveys on the prevalence of contamination of cattle with verocytotoxin-producing E. coli (VTEC) serogroup O157:H7. These surveys have been conducted since 1986 on farms in North America (10 studies), on farms in Europe (6 studies) and at slaughterhouses prior to or just after slaughter (7 studies) or after skinning and evisceration (3 studies). The purpose of this review is to understand the first stages of the epidemiology of the infection in animals and humans (the infection process being obscure in many points) and to prepare herd-based control measures to reduce the risk of O157:H7 human infection. The different statistical methods employed in these surveys, as well as the various laboratory screening methods used for detecting positive animals are presented. The observed frequencies of infected animals (animal prevalence) and herds (herd prevalence) are given as a function of localisation, year, type of industry (beef or dairy) and age. From these measured prevalence values, the risk of contamination of ground beef by E. coli O157:H7 in the first stages of the farm-to-fork continuum is assessed. First, we follow the evolution of contamination frequencies from the living animal on-farm to carcasses before transformation. Then, within each set of measurements (i.e., on farm or at slaughterhouse), we identify the effects of the following factors: target population, sampling strategies and laboratory procedures. We argue that the prevalence values inferred from these measurements are very likely underestimated, due to insufficient sampling and not enough sensitive laboratory procedures (one exception being the immunomagnetic bead separation technique). No firm conclusion can be drawn as to the effects of geographical localisation and season. In those surveys, the effect of hygiene level at slaughterhouse on prevalence values is not quantitatively assessed. In addition, there is growing evidence of other sources of E. coli O157:H7 than live cattle in the farm environment, such as feed, water and water-troughs.     

Prevalence of Escherichia coli O157:H7 in range beef calves at weaning

This study was designed to determine the prevalence of Escherichia coli O157:H7 infection of beef calves at weaning, prior to arrival at the feedlot or mixing with cattle from other sources. Fifteen range cow-calf herds, which weaned calves in October and November, were sampled in Kansas, Missouri, Montana, Nebraska and South Dakota. Faecal culture for E. coli O157:H7 was performed and anti-O157 serum antibody titres were determined by blocking ELISA. Thirteen of the 15 herds (87%) were found to have at least one positive isolation of E. coli O157:H7 in faecal samples. Within positive herds, prevalence ranged from 1.7-20.0%, with an average of 7.4+/-6.2% S.D. of individual animals shedding E. coli O157:H7 in faeces. All herds had high prevalence of anti-O157 antibodies, ranging 63-100% of individuals within herds seropositive. This study indicates that E. coli O157:H7 infection before weaning, prior to entry into feedlots, is widespread. Furthermore, serologic evidence suggests that most calves (83%) and all herds (100%) have been exposed to E. coli O157.     

The prevalence of Escherichia coli O157.H7 in dairy and beef cattle in Washington State.

Escherichia coli O157.H7 was found in 10 of 3570 (0.28%) faecal samples from dairy cattle in 5 of 60 herds (8.3%). Several tentative associations with manure handling and feeding management practices on dairy farms were identified. Faecal/urine slurry samples, bulk milk samples, and milk filters from dairy herds were negative for E. coli O157.H7. E. coli O157.H7 was also isolated from 10 of 1412 (0.71%) faecal samples from pastured beef cattle in 4 of 25 (16%) herds. The prevalence of E. coli O157.H7 excretion in feedlot beef cattle was 2 of 600 (0.33%). The identification of cattle management practices associated with colonization of cattle by E. coli O157.H7 suggests the possibility that human E. coli O157.H7 exposure may be reduced by cattle management procedures.     

The role of heightened surveillance in an outbreak of Escherichia coli O157.H7.

After instituting laboratory screening for Escherichia coli O157.H7, a Connecticut hospital isolated the organism from four persons in September 1993. As a result, an outbreak of E. coli O157.H7 associated with a country club was detected. The club had served hamburger from the same shipment at two picnics. Attendees of two picnics were interviewed, stool cultures were obtained from symptomatic persons, and the remaining hamburger was cultured. Twenty (22%) of 89 persons who ate hamburger became ill, compared with 1 of 60 who did not eat hamburger (relative risk = 13.5, 95% confidence interval 3.2-56.3). Among persons who ate hamburgers, illness was strongly associated with eating hamburger that was not thoroughly cooked (P < 0.001). All 20 samples from 5 remaining boxes of patties yielded E. coli O157.H7. Isolates from hamburger and case-patients were indistinguishable by pulsed-field gel electrophoresis. Heightened surveillance can rapidly identify outbreaks and may mitigate their impact. However, continued review of food safety issues is necessary if E. coli O157.H7 outbreaks are to be prevented.     

An outbreak of Escherichia coli O157:H7 infections and haemolytic uraemic syndrome associated with consumption of unpasteurized apple cider

During October 1996, an outbreak of Escherichia coli O157:H7 infections among Connecticut residents occurred. An epidemiologic investigation included enhanced surveillance and a case-control study. Clinical isolates of Escherichia coli O157:H7 were typed by pulsed-field gel electrophoresis (PFGE). Implicated cider samples were analysed by culture and polymerase chain reaction (PCR). Consumption of implicated cider was associated with illness; (matched odds ratio = undefined, 95 % confidence interval = 3.5-infinity). Ultimately, a total of 14 outbreak-associated patients were identified. All isolates analysed by PFGE yielded the outbreak-associated subtype. Escherichia coli O157:H7 was not cultured from three cider samples; PCR analysis detected DNA fragments consistent with Escherichia coli O157:H7 in one. This outbreak was associated with drinking one brand of unpasteurized apple cider. PFGE subtyping supported the epidemiologic association. PCR analysis detected microbial contaminants in the absence of live organisms. Washing and brushing apples did not prevent cider contamination.     

Surveillance of Escherichia coli O157 isolation and confirmation, United States, 1988

In 1989, to examine patterns of testing for Escherichia coli O157:H7 in state public health laboratories (SPHLs), CDC conducted a survey to determine the availability and type of Escherichia coli O157:H7 testing in SPHLs during 1988 and the number of isolates confirmed at SPHLs if such testing was available. The results were compared with information on isolates submitted for confirmatory testing at CDC in 1988. Thirty-nine (78%) of the 51 SPHLs were testing for E. coli O157:H7 in 1988; 26 confirmed at least one E. coli O157 isolate in that year. CDC confirmed isolates from three additional states. A total of 489 E. coli O157:H7 or E. coli O157:NM isolates were identified, with the largest numbers being reported from Washington (156), Oregon (64), Minnesota (63), and Massachusetts (36). These results show that E. coli O157 has been detected in most areas of the United States. Infections are apparently concentrated in northern states; however, improved surveillance data are needed to determine regional incidence and trends.     

Beef patty composition: effects of fat content and cooking method

Patties from ground round, ground chuck , and regular ground beef containing 14%, 19%, and 24% fat, respectively, were cooked by six different methods and analyzed for compositional differences. Cooking methods were electric broiling , charbroiling , roasting, convection heating, frying, and microwave heating. Percent yield, fat, and moisture of cooked patties were significantly affected (p less than .05) by the interaction of fat level and cooking method. Except for patties cooked by broiling or convection heating, patty yields decreased with increasing fat levels. Leaner formulations of ground beef increased in fat percentage with cooking, while patties processed to contain more fat initially decreased in fat percentage following cooking. Microwave cooking always produced patties containing the least fat and caloric content in comparison with other methods. Increasing levels of fat in the raw patty formulation did not appear to affect substantially the total caloric value of the patties following cooking. Cooked patties processed to have 14% fat contained slightly more water and less fat than patties having 24% fat. At current retail prices, cooked patties made to have 14% fat cost +0.20 more per patty than patties produced to contain 24% fat, regardless of the method of cooking.     

Prevalence of verocytotoxigenic Escherichia coli in ground beef, pork, and chicken in southwestern Ontario

Samples of ground beef (225), pork (235) and chicken (200) were randomly selected from meat processing plants in the southwestern Ontario area. Supernatants of broth cultures of the samples were tested for verocytotoxins using a Vero cell assay. Neutralization of cytotoxic activity using antisera specific for three types of verocytotoxin (Verotoxin 1, Verotoxin 2 and Shiga-like toxin II) was performed on positive samples. Isolation of verocytotoxigenic Escherichia coli (VTEC) was attempted from positive samples. VTEC were confirmed as E. coli biochemically, tested for drug resistance, and serotyped. Based on neutralization studies, the prevalence of VTEC in beef and pork was at least 36.4% and 10.6%, respectively. This is much higher than has been reported from a survey of retail meats in which a method designed to detect only E. coli O 157.H7 was used. Isolations of VTEC were made from 10.4% of the beef samples and 3.8% of the pork samples. No VTEC were recovered from the chicken samples. The majority of VTEC isolates were susceptible to commonly used antimicrobial agents. A number of the serotypes of the VTEC isolates recovered have been associated with human disease; however, no VTEC of serotype O 157.H7 were isolated.