Escherichia coli (STEC) serotype O104 outbreak causing haemolytic syndrome (HUS) in Germany and France

At the beginning of May an outbreak of bloody diarrhoea and haemolytic uraemic syndrome (HUS) began in Germany. During the succeeding months following the initial outbreak in Germany, thousands of infections occurred resulting in 877 cases of haemolytic uraemic syndrome (HUS) with 32 deaths and 3,043 cases of enterohaemorrhagic Escherichia coli (EHEC) with 16 deaths.     

Enteroaggregative E. coli O104 from an outbreak of HUS in Germany 2011, could it happen again?

Enterohaemorrhagic E. coli (EHEC) particularly O157:H7 (Sequence type 11 complex), is the best documented and most well-known of E. coli that cause diarrhoea. The importance of EHEC lies in the severity of disease. Outbreaks can infect thousands of people causing bloody diarrhoea and haemolytic uremic syndrome (HUS) that in turn can result in protracted illness or even death. The ability of EHEC to colonise the human gut is normally associated with the presence of genes from another group of diarrhoeagenic E. coli, the enteropathogenic E. coli (EPEC), via the locus of enterocyte effacement. However, the massive outbreak in Germany was caused by an EHEC which had acquired virulence genes from yet another group of diarrhoeagenic E. coli, the enteroaggregative E. coli (EAEC). In reality EAEC is probably the most common bacterial cause of diarrhoea but is not identified in most diagnostic laboratories. This outbreak emphasises the importance of being able to detect all diarrhoeagenic E. coli and not to focus on E. coli O157:H7 alone. Routine surveillance systems for EAEC, a once ignored global pathogen, would go a long way to reaching this goal. This review describes methods for identifying non-O157 EHEC and describes the key genetic features of EHEC and EAEC. Our aim is to provide information for laboratories and policy makers which enables them to make informed decisions about the best methods available for detecting newly emergent strains of diarrhoeagenic E. coli.     

肠出血性大肠杆菌O104:H4病原学及其检测方法研究进展

2011年5月以来,德国暴发了由肠出血性大肠杆菌O104:H4感染引起的疫情,世界多个国家相继报告有此类病例发生,这是首次报告由肠出血性大肠杆菌O104:H4感染引起疾病的暴发流行。截至2011年6月24日,已造成3920例感染和48例死亡[1]。该病患者以血水样腹泻、呕吐起病,部分病例并发溶血性尿毒综合征,导致多器官受损,甚至死亡。疫情发生后,世界各国相关学者立即开展其病原学特性研究,同时迅速研制出实验室检测方法和试剂,为该病的及时诊断、治疗和疫情控制发挥了很大作用。本文介绍了肠出血性大肠杆菌O104:H4病原学及其检测方法的研究进展。     

Clinical management of children with suspected or confirmed E. coli O157 infection

Scotland continues to report higher rates of infection with Escherichia coli O157 than elsewhere in the UK. Infection with E. coli O157 usually manifests as acute, afebrile, painful, bloody diarrhoea and is the commonest cause of Haemolytic Uraemic Syndrome (HUS), an important cause of childhood renal failure. In 1996 an outbreak of E. coli O157 infection in Central Scotland, resulted in over 500 cases and 17 deaths. Ten years on, high-profile outbreaks of E. coli O157 infection in Scotland still result in cases of HUS and fatalities. We sought to identify outcomes and describe pre-hospital clinical management strategies using prospective, national surveillance of paediatric HUS cases, from 2003 to 2006 inclusive. We recommend that children who present with acute, afebrile, and painful bloody diarrhoea be referred to hospital as early as possible for appropriate clinical management.     

Clonal diversity of Shiga toxin-producing Escherichia coli O103:H2/H(-) in Germany.

Shiga toxin producing Escherichia coli O103:H2/H(-) belong to the third most frequently isolated EHEC serotypes in Germany following isolates of O157:H7/H(-) and O26:H11/H(-). A total of 145 respective E. coli 103 isolates from single cases of diarrhoea and haemolytic uremic syndrome (HUS) in 1997-2000 were characterised by a range of molecular subtyping methods (PFGE, P-gene profiling, ribotyping, electrotyping) and phage typing in order to analyse their genetic relatedness and the practicability for new epidemiological tracing back. All isolates cluster into a distinct EHEC subgroup and reveal a high clonal diversity together with a considerable stability. Since strains of this serotype rank up to the third most frequently isolated EHEC in Germany a large population of this serotype, and therefore, a great supply of such strains may exist in this country.     

An outbreak of Escherichia coli O157 gastroenteritis in a care home for the elderly

In the summer of 2001 an outbreak of Escherichia coli O157 gastroenteritis affected staff and residents of a care home for the elderly in the West Midlands, UK. E. coli O157 phage type 2 was isolated from faeces in eight patients and 12 staff members. Thirty-five staff and 40 residents met the case definition for clinical gastrointestinal infection. Serological testing identified a further 14 possible cases of infection amongst asymptomatic staff and residents. The outbreak was atypical, as the disease seemed to be milder than has been observed in past outbreaks in similar settings. The index case, a member of staff, developed bloody diarrhoea and haemolytic-uraemic syndrome (HUS), but only one resident developed bloody diarrhoea and required hospitalization. No deaths occurred, despite the high-risk nature of the affected population. The source of the outbreak could not be identified. The prolonged nature of the outbreak and observed lapses in infection control practices indicated that person-to-person spread was the likely route of transmission. This outbreak illustrates the importance of observing appropriate infection control measures in the institutions providing residential and nursing care to the elderly.     

Investigation of human infections with verocytotoxin-producing strains of Escherichia coli (VTEC) belonging to serogroup O118 with evidence for zoonotic transmission

Twenty verocytotoxigenic Escherichia coli (VTEC) O118 strains isolated between 1996 and 1998 from human patients in Germany were analysed for their serotypes, their virulence markers and their epidemiological relatedness. Three strains were typed as O118:H12, these carried only the VT2d-Ount variant gene and were not associated with diarrhoea or haemolytic uraemic syndrome (HUS). Seventeen strains were serotyped as O118:H16 or O118:non-motile (NM). These carried all the genes for VTI, eae and EHEC-haemolysin. The O118:H16/NM strains were from diarrhoea (13 cases) and HUS (2 cases). Sixteen of the patients were young infants and most infections were associated with a rural environment. Evidence for zoonotic transmission from cattle to humans was found in two cases. The epidemiological relationship between the human and bovine O118:H16/NM isolates was indicated by homogeneous plasmid patterns and by very similar XbaI restriction patterns obtained by pulsed-field gel electrophoresis. VTEC O118:H16/NM are emerging pathogens in Germany and should be classified as new enterohaemorrhagic E. coli (EHEC) types.     

肠出血性大肠埃希菌感染的流行病学及临床诊治新进展

肠出血性大肠埃希菌(EHEC)是一种高度危险的致病菌,主要引起出血性肠炎,但有少部分患者感染后可能发展为威胁生命的疾病,例如溶血尿毒症综合征(HUS),而HUS是欧美儿童急性肾功能衰竭最常见的原因.绝大多数EHEC感染是可以预防的,一旦EHEC感染发生,目前还没有任何有效的措施预防HUS的发生.此文主要介绍了EHEC流...     

The stimulating effect of fosfomycin,an antibiotic in common use in Japan,on the production/release of verotoxin-1 from enterohaemorrhagic Escherichia coli O157:H7 in vitro.

In 1996 Japan had a big outbreak of enterohaemorrhagic Escherichia coli (EHEC) O157:H7, especially in Osaka prefecture where about 6000 patients (mainly schoolchildren) suffered from diarrhoea, haemorrhagic colitis and haemolytic uraemic syndrome (HUS) due to EHEC infection via school lunch. Our survey of doctors who took care of those patients showed that most patients had received antibiotics, especially fosfomycin which comprised 84% of the prescribed treatment. Since the administration of antibiotics for EHEC infection is under discussion, we analysed the effect of fosfomycin on production/release of verotoxins (VTs). Addition of fosfomycin into EHEC culture in CAYE broth at 2 and 5 h after start of incubation caused marked increase of VT1 release. This observation warns of the possibility of fosfomycin treatment making the disease worse.     

Prevention and control of enterohaemorrhagic Escherichia coli (EHEC) infections: memorandum from a WHO meeting. WHO Consultation on Prevention and Control of Enterohaemorrhagic Escherichia coli (EHEC) Infections.

Escherichia coli is a commonly occurring inhabitant of the intestine of humans and other animals, but there are several pathogenic types of E. coli which cause a variety of human diseases. One of these pathogenic types, E. coli O157:H7, belongs to the group of enterohaemorrhagic E. coli (EHEC) which produce potent toxins and cause a particularly severe form of disease, haemorrhagic colitis (HC). About 10% of patients with HC can go on to develop haemolytic uraemic syndrome (HUS), a life-threatening complication of E. coli O157:H7 infection that is characterized by acute renal failure, haemolytic anaemia, and thrombocytopenia. These sequelae are particularly serious in young children and older people. On average, 2-7% of patients with HUS die, but in some outbreaks among the elderly the mortality rate has been as high as 50%. This Memorandum reviews the growing importance of E. coli O157:H7 as a foodborne pathogen and reports on the issues of surveillance, outbreak investigation, and control strategies with respect to EHEC infections that were discussed at the WHO Consultation on Prevention and Control of EHEC Infections, held in Geneva on 28 April to 1 May 1997. Recommended measures for prevention and control include the following: use of potable water in food production; presentation of clean animals at slaughter; improved hygiene throughout the slaughter process; appropriate use of food processing measures; thorough cooking of foods; and the education of food handlers, abattoir workers, and farm workers on the principles and application of food hygiene.     

Escherichia coli O104:H4 outbreak from sprouted seeds

From May to July 2011, one of the largest reported outbreaks of haemolytic uraemic syndrome (HUS) and bloody diarrhoea caused by the Shiga toxin-producing Escherichia coli (STEC) O104:H4 occurred in Germany and France. The hypothetical origin of the outbreak strain was a combined enteroaggregative E. coli and an enterohaemorrhagic E. coli with the ability to resist multi-antibiotics and produce Shiga-toxin 2. The combination of aggregative ability, antibiotic resistance and the production of Shiga-toxin 2 significantly affected the severity of the symptoms presented. Since humans may be the primary reservoir, it is likely that contamination could have occurred through contact with infected individuals. Farm food safety management, and hand hygiene training programmes are crucial to primary production to prevent or reduce risks of contamination.     

Vero cytotoxin-producing Escherichia coli, particularly serogroup O 157, associated with human infections in the United Kingdom: 1989-91.

This survey reports the results of investigations performed by the Laboratory of Enteric Pathogens (LEP), to identify evidence of human infection with Vero cytotoxin-producing Escherichia coli (VTEC) in the UK during the period 1989-91. Bacterial isolates, faecal specimens and serum samples were received from patients suffering from diarrhoea, bloody diarrhoea and haemolytic uraemic syndrome. Using serotyping, Vero cytotoxin gene probing and an ELISA for serum antibodies to E. coli O 157, evidence of infection was detected in 232, 428 and 615 individuals in 1989, 1990 and 1991 respectively. Of these individuals, 15% were reported as having HUS. Vero cytotoxin-producing E. coli O 157 was the most frequently encountered serogroup, with isolations from a total of 1092 individuals over the 3-year period. The incidence of VTEC infection increased from 0.41/100,000 in 1989 to 1.07/100,000 in 1991. The area with the highest rate of infection in each year was Scotland, increasing from 1.37/100,000 in 1989 to 3.97/100,000 in 1991.     

An international outbreak of Vero cytotoxin-producing Escherichia coli O157 infection amongst tourists; a challenge for the European infectious disease surveillance network

In March 1997, an outbreak of Vero cytotoxin-producing Escherichia coli O157 (VTEC) infection occurred amongst holidaymakers returning from Fuerteventura, Canary Islands. For the investigation, a confirmed case was an individual staying in Fuerteventura during March 1997, with either E. coli O157 VTEC isolated in stool, HUS or serological evidence of recent infection; a probable case was an individual with bloody diarrhoea without laboratory confirmation. Local and Europe-wide active case finding was undertaken through national centres, Salm-Net and the European Programme of Intervention Epidemiology, followed by a case-control study. Fourteen confirmed and one probable case were identified from England (7), Finland (5), Wales (1), Sweden (1) and Denmark (1) staying in four hotels. Three of the four hotels were supplied with water from a private well which appeared to be the probable vehicle of transmission. The case-control study showed illness was associated with consumption of raw vegetables (OR 8.4, 95% CI 1-5-48.2) which may have been washed in well water. This investigation shows the importance of international collaboration in the detection and investigation of clusters of enteric infection.     

Epidemiology of infection due to Escherichia coli O157: a 3-year prospective study.

A 3-year study of Escherichia coli infections in Grampian Region was conducted to ascertain the incidence, document clinical sequelae and identify at-risk groups. Approximately 30,000 stools from patients with acute diarrhoea were screened for E. coli O157, and an epidemiological questionnaire filled in for each patient whose stool was positive. Eighty-three patients were studied. The annual incidence was 6 per 100, 000. Proportionately more infections occurred in people involved in agriculture. Evidence was seen of case-to-case transmission, and contamination of a water supply. Eight cases developed haemolytic uraemic syndrome (HUS). There were 2 deaths due to HUS and 2 due to haemorrhagic colitis (HC). Symptomatic E. coli infection is relatively common in the Grampian Region, more common in the agricultural community, and is the main cause of HUS in this Region.     

Three years experience of adults admitted to hospital in north-east Scotland with E. coli O157

To describe the epidemiology, clinical features, treatment and outcomes of adults with E. coli O157 infection presenting to Aberdeen Royal Infirmary over a three year period. METHODS: A retrospective casenote review. RESULTS: Thirty-two confirmed cases of E. coli O157 infection were admitted between 1997 and 2000. The median age was 58 years (range 16-93). Ten patients (31%) were from the city of Aberdeen and 22 (69%) from surrounding rural areas. Twenty-seven patients (85%) presented between May and October. The source of infection was unknown or unconfirmed in all cases. Bloody diarrhoea was present in 30 (94%). Leucocytosis was present in 18 (63%) but only four patients (13%) had a fever. Six of the 32 patients (19%) developed Haemolytic-Uraemic Syndrome (HUS) of whom 2 died. Ten patients received antibiotics of whom two developed HUS. Twenty-seven of the 32 (85%) had made a full recovery by time of discharge, three (9%) had impaired renal function and two (6%) died in hospital. CONCLUSION: E. coli O157 infection tends to occur sporadically in rural areas in North East Scotland. It is not usually associated with fever. Infection occurs more commonly in the summer and autumn. HUS complicates infection in almost one fifth of patients.     

Predictors for the development of haemolytic uraemic syndrome with Escherichia coli O157:H7 infections: with focus on the day of illness

A large outbreak of Escherichia coli O157 infections via school lunches occurred at primary schools in 1996 in Sakai City, Japan. As many as 10,000 patients suffered from diarrhoea, haemorrhagic colitis and haemolytic uraemic syndrome (HUS). Using data on 288 inpatient school children affected by this outbreak, of whom 36 presented complete HUS and the remaining 252 tested positive for E. coli O157 culture, we attempted to identify predictors for the progression to HUS. Within the first 5 days of illness, clinical features associated with inpatients who developed HUS compared with those without HUS included a C reactive protein (CRP) level higher than 1.2 mg/dl (OR 44.26; 95% CI 5.83-336.23), a white blood cell (WBC) count greater than 11.0 x 10(9)/l (OR 5.03; 95% CI 2.13-11.87) and a temperature higher than 38.0 degrees C (OR 5.00; 95% CI 2.25-11.08). It can be concluded that these three factors are predictive factors for the development of HUS in patients with E. coli O157 infection, and patients who have two or all of these factors should be observed closely.     

Sequence heterogeneity of the eae gene and detection of verotoxin-producing Escherichia coli using serotype-specific primers.

The distribution of the Escherichia coli attaching and effacing (eae) gene in strains of verotoxin-producing E. coli (VTEC) isolated from cattle and humans was studied. The majority of strains isolated from humans with bloody diarrhoea or HUS and cattle with severe diarrhoea were eae positive (82 and 83% respectively). In contrast, 59% of VTEC isolated from asymptomatic cattle were eae negative and of the remaining 41% that were eae positive, the majority were serotype O157. H7. The nucleotide sequence of the 3'' end of the eae gene of enteropathogenic E. coli (EPEC) of serotype O55. H7 was found to be almost identical to that of serotype O157. H7. Specific primers are described which detect the eae sequences of VTEC serotypes O157. H7, O157. H-, and EPEC serotypes O55. H7 and O55. H-. The nucleotide sequence of the 3'' end of the eae gene of serotype O111. H8 differed significantly from that of O157. H7. Primers were developed to specifically identify the eae sequences of VTEC serotypes O111. H- and O111. H8. We conclude that whereas the majority of VTEC associated with disease in cattle and humans possess the eae gene, the gene itself may not be necessary to produce haemorrhagic colitis and HUS. Sequence heterogeneity in the 3'' end of eae alleles of VTEC permits specific identification of subsets of these organisms.     

Epidemiological analysis of a cluster within the outbreak of Shiga toxin-producing Escherichia coli serotype O104:H4 in Northern Germany, 2011

In May 2011 one of the worldwide largest outbreaks of haemolytic uraemic syndrome (HUS) and bloody diarrhoea caused by Shiga toxin-producing Escherichia coli (STEC) serotype O104:H4 occurred in Germany. One of the most affected federal states was Lower Saxony. We present the investigation of a cluster of STEC and HUS cases within this outbreak by means of a retrospective cohort study. After a 70th birthday celebration which took place on 7th of May 2011 among 72 attendants seven confirmed cases and four probable cases were identified, two of them developed HUS. Median incubation period was 10 days. Only 35 persons (48.6%) definitely answered the question whether they had eaten the sprouts that were used for garnishing the salad. Univariable analysis revealed different food items, depending on the case definition, with Odds Ratio (OR) > 1 indicating an association with STEC infection, but multivariable logistic regression showed no increased risk for STEC infection for any food item and any case definition. Sprouts as the source for the infection had to be assumed based on the results of a tracing back of the delivery ways from the catering company to the sprouts producer who was finally identified as the source of the entire German outbreak. In this large outbreak several case–control studies failed to identify the source of infection.     

Diarrhoea in close contacts as a risk factor for childhood haemolytic uraemic syndrome. The CPKDRC co-investigators.

To determine whether the risk factors for childhood haemolytic uraemic syndrome (HUS) are similar to risk factors previously reported for Escherichia coli O 157. H7 gastroenteritis, we conducted a case-control study at eight paediatric hospitals in the summer of 1990. Thirty-four consecutive children with HUS were prospectively enrolled; all had diarrhoea and 88% had laboratory evidence of exposure to verotoxin-producing E. coli (VTEC). The 102 controls were otherwise healthy children with minor acute injuries. Parents of all subjects responded to a questionnaire about each child''s exposure to various foods, methods of food preparation, sources of water, travel, and individuals with diarrhoea. Children with HUS were significantly more likely than controls to have had close contact with an individual with diarrhoea in the 2 weeks before the onset of illness (74 v. 29%, P < 0.00001; odds ratio 7.0, 95% CI 2.7-18.5). The onset of diarrhoea in the contacts occurred a median of 6 days (range, 1- > 14 days) before the onset of diarrhoea in the HUS patients. Exposure to undercooked ground meat was not significantly more common in the patients with HUS (15 v. 8%; P = 0.05). These data provide evidence consistent with person-to-person transmission of VTEC in a substantial proportion of episodes of childhood HUS.     

An outbreak of infection due to verocytotoxin-producing Escherichia coli O157 in four families: the influence of laboratory methods on the outcome of the investigation.

Three members of family A, who had diarrhoea on 20 October, lived on a small arable farm which had 10 cattle. Manure from the animals was used to fertilize the ground for growing potatoes which were then offered for retail sale, unwashed, directly from the farm. The mother from family B bought potatoes, which were covered with manure, from family A in early November and over the subsequent 10 days she became ill with diarrhoea and her daughter and son both became ill with bloody diarrhoea. The mother from family C visited family B while the daughter from the latter family was symptomatic; the mother developed diarrhoea several days later. The mother and two sons from family D visited family B while the son from the latter family was symptomatic; the first son developed bloody diarrhoea 6 days later which progressed to development of haemolytic-uraemic syndrome. Direct culture of faecal samples onto cefixime rhamnose sorbitol MacConkey agar failed to isolate E. coli O157 from any of the symptomatic patients, and direct culture onto cefixime tellurite sorbitol MacConkey agar isolated the organism from only one patient. In contrast, a combination of isolation of E. coli O157 by immunomagnetic separation and detection of E. coli O157-specific secretory IgA, suggested E. coli O157 infection in all eight symptomatic patients, but not in any of the family members who were not ill. Two children who excreted the organism for 60 and 89 days respectively were the only two patients who did not develop a secretory IgA response. E. coli O157 was not isolated from potatoes from the farm and faecal samples from the farm animals were not available for examination. The study illustrates the need to use the most sensitive methods available during the investigation and follow up of cases of E. coli O157 infection.