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Assessment of Shiga toxin-producing

Escherichia coli isolates from wildlife

meat as potential pathogens for
Miko A, Pries K, Haby S, Steege K, Albrecht N, Krause G, Beutin L.

National Reference Laboratory for Escherichia coli, Federal Institute for Risk
Assessment (BfR), Diedersdorfer Weg 1, D-12277 Berlin, Germany.


A total of 140 Shiga toxin-producing Escherichia coli (STEC) strains from wildlife meat
(deer, wild boar, and hare) isolated in Germany between 1998 and 2006 were
characterized with respect to their serotypes and virulence markers associated with
human pathogenicity. The strains grouped into 38 serotypes, but eight O groups (21, 146,
128, 113, 22, 88, 6, and 91) and four H types (21, 28, 2, and 8) accounted for 71.4% and
75.7% of all STEC strains from game, respectively. Eighteen of the serotypes, including
enterohemorrhagic E. coli (EHEC) O26:[H11] and O103:H2, were previously found to be
associated with human illness. Genes linked to high-level virulence for humans (stx(2),
stx(2d), and eae) were present in 46 (32.8%) STEC strains from game. Fifty-four STEC
isolates from game belonged to serotypes which are frequently found in human patients
(O103:H2, O26:H11, O113:H21, O91:H21, O128:H2, O146:H21, and O146:H28). These
54 STEC isolates were compared with 101 STEC isolates belonging to the same
serotypes isolated from farm animals, from their food products, and from human patients.
Within a given serotype, most STEC strains were similar with respect to their stx
genotypes and other virulence attributes, regardless of origin. The 155 STEC strains were
analyzed for genetic similarity by XbaI pulsed-field gel electrophoresis. O103:H2,
O26:H11, O113:H21, O128:H2, and O146:H28 STEC isolates from game were 85 to
100% similar to STEC isolates of the same strains from human patients. By multilocus
sequence typing, game EHEC O103:H2 strains were attributed to a clonal lineage
associated with hemorrhagic diseases in humans. The results from our study indicate that
game animals represent a reservoir for and a potential source of human pathogenic STEC
and EHEC strains.

PMID: 19700552 [PubMed - indexed for MEDLINE]PMCID: PMC2765146Free PMC



Methods for the detection and isolation of
Shiga toxin-producing Escherichia coli.
De Boer E, Heuvelink AE.

Inspectorate for Health Protection, Zutphen, The Netherlands.


Shiga toxin-producing Escherichia coli (STEC) are an important cause of haemorrhagic

colitis and the diarrhoea-associated form of the haemolytic uraemic syndrome. Of the
numerous serotypes of E. coli that have been shown to produce Shiga toxin (Stx), E. coli
O157:H7 and E. coli O157:NM (non-motile) are most frequently implicated in human
disease. Early recognition of STEC infections is critical for effective treatment of
patients. Furthermore, rapid microbiological diagnosis of individual patients enables the
prompt notification of outbreaks and implementation of control measures to prevent more
cases. Most human infections caused by STEC have been acquired by the consumption of
contaminated foods, especially those of bovine origin such as undercooked ground beef
and unpasteurized cows' milk, and by person-to-person contacts. To identify the
reservoirs of STEC and the routes of transmission to man, sensitive methods are needed
as these pathogens may only be present in food, environmental and faecal samples in
small numbers. In addition, sensitive and rapid detection methods are necessary for the
food industry to ensure a safe supply of foods. Sensitive methods are also needed for
surveillance programmes in risk assessment studies, and for studies on survival and
growth of STEC strains. Cultural methods for the enrichment, isolation and confirmation
of O157 STEC are still evolving. Several selective enrichment media have been
described, of which modified tryptone soy broth with novobiocin and modified E. coli
broth with novobiocin, seem to be the most appropriate. These media are minimally-
selective broths that give a somewhat limited differential specificity favouring isolation
of O157 STEC, as opposed to other Gram-negative bacteria, in the sample. An incubation
temperature of 41-42 degrees C further enhances selectivity. The occurrence of heat-,
freeze-, acid- or salt-stressed STEC in foods means that it is important to be able to detect
cells that are in a stressed state, as STEC generally have a very low infectious dose, and
injured cells mostly retain their pathogenic properties. For the isolation of stressed O157
STEC, pre-enrichment in a non-selective broth is necessary. The most widely used
plating medium for the isolation of typical sorbitol-non-fermenting strains of STEC of
serogroup O157 is sorbitol MacConkey agar with cefixime and tellurite (CT-SMAC). As
some STEC strains are sensitive for tellurite and/or are sorbitol-fermenting, the use of a
second isolation medium, such as one of the newer chromogenic media, is recommended.
Immunomagnetic separation (IMS) following selective enrichment, and subsequent
spread-plating of the concentrated target cells onto CT-SMAC agar, appears to be the
most sensitive and cost-effective method for the isolation of E. coli O157 from raw foods.
IMS increases sensitivity by concentrating E. coli O157 relative to background
microflora, which may overgrow or mimic O157 STEC cells on selective agars. While
cultural isolation of O157 STEC from foods and faeces is time-consuming, labour-
intensive and hence, costly, rapid immunological detection systems have been developed
which significantly reduce the analysis time. These methods include enzyme-linked
immunosorbent assays (ELISAs), colony immunoblot assays, direct immunofluorescent
filter techniques, and several immunocapture techniques. Both polyclonal and
monoclonal antibodies specific for the O and H antigens are used for these methods.
Many of these test systems are able to detect less than one O157 STEC cell g(-1) of raw
meat after overnight enrichment. Presumptive results are available after just one day, but
need to be completed with the isolation of the organisms. The primary use of these
procedures is therefore to identify food and faecal samples that possibly contain O157

PMID: 10880188 [PubMed - indexed for MEDLINE]



Prevalence and pathogenicity of Shiga

toxin-producing Escherichia coli in beef
cattle and their products.
Hussein HS.

Department of Animal Biotechnology, University of Nevada, Reno 89557, USA.



During the past 23 yr, a large number of human illness outbreaks have been traced
worldwide to consumption of undercooked ground beef and other beef products
contaminated with Shiga toxin-producing Escherichia coli (STEC). Although several
routes exist for human infection with STEC, beef remains a main source. Thus, beef
cattle are considered reservoirs of O157 and nonO157 STEC. Because of the global
nature of the food supply, safety concerns with beef will continue, and the challenges
facing the beef industry will increase at the production and processing levels. To be
prepared to address these concerns and challenges, it is critical to assess the beef cattle
role in human infection with STEC. Because most STEC outbreaks in the United States
were traced to beef containing E. coli O157:H7, the epidemiological studies have focused
on the prevalence of this serotype in beef and beef cattle. Worldwide, however, additional
STEC serotypes (e.g., members of the O26, O91, O103, O111, O118, O145, and O166
serogroups) have been isolated from beef and caused human illnesses ranging from
bloody diarrhea and hemorrhagic colitis to the life-threatening hemolytic uremic
syndrome (HUS). To provide a global assessment of the STEC problem, published
reports on beef and beef cattle in the past 3 decades were evaluated. The prevalence rates
of E. coli O157 ranged from 0.1 to 54.2% in ground beef, from 0.1 to 4.4% in sausage,
from 1.1 to 36.0% in various retail cuts, and from 0.01 to 43.4% in whole carcasses. The
corresponding prevalence rates of nonO157 STEC were 2.4 to 30.0%, 17.0 to 49.2%,
11.4 to 49.6%, and 1.7 to 58.0%, respectively. Of the 162 STEC serotypes isolated from
beef products, 43 were detected in HUS patients and 36 are known to cause other human
illnesses. With regard to beef cattle, the prevalence rates of E. coli O157 ranged from 0.3
to 19.7% in feedlots and from 0.7 to 27.3% on pasture. The corresponding prevalence
rates of nonO157 STEC were 4.6 to 55.9% and 4.7 to 44.8%, respectively. Of the 373
STEC serotypes isolated from cattle feces or hides, 65 were detected in HUS patients and
62 are known to cause other human illnesses. The results indicated the prevalence of a
large number of pathogenic STEC in beef and beef cattle at high rates and emphasized
the critical need for control measures to assure beef safety.

PMID: 17060419 [PubMed - indexed for MEDLINE]Free Article

Publication Types, MeSH Terms, Substances

Publication Types:

• Research Support, U.S. Gov't, Non-P.H.S.

• Review

MeSH Terms:

• Animals
• Cattle/microbiology
• Cattle Diseases/epidemiology*
• Cattle Diseases/microbiology
• Disease Outbreaks
• Disease Reservoirs*
• Escherichia coli/isolation & purification
• Escherichia coli/pathogenicity*
• Escherichia coli Infections/epidemiology
• Escherichia coli Infections/microbiology
• Escherichia coli Infections/veterinary*
• Escherichia coli O157/isolation & purification
• Escherichia coli O157/pathogenicity*
• Humans
• Meat/microbiology*
• Prevalence
• Shiga Toxins/biosynthesis*


• Shiga Toxins

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