Shiga Toxin Producing Escherichia coli: Recent Trends and Facts

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E. coli O157:H7 Nomenclature Introduction Outbreaks Non- 0157 STEC Animal Reservoir Transmission Antibiotic Resistance Detection Methods Summary

 Gram negative rod shaped bacterium  Facultative anaerobe  Flagella are encoded by the H7 antigen  Produces Shigatoxin

.. Nebraska) ƒ ƒ Verocytotoxin-producing E. coli (VTEC) Shiga toxin-producing E. Clay Center.Lipopolysaccharide (LPS) = O antigen O1-O181 Y Flagella = H antigen H1-H56 Y (USDA-ARS. 1977) (O Brien et al. 1983) . coli (STEC) (Konawalchuck et al.

coli O157:H7 .ƒ STEC cause a serious illness with symptoms of severe bloody diarrhea and kidney failure hemolytic uremic syndrome (HUS) ƒ The STEC serotype that causes the most outbreaks and cases of HUS in the United States is E.

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each year ƒ E.S.ƒ According to the CDC.000 nonO157:H7 STEC infections occur in U. coli O157:H7 infections and at least 37. coli O157:H7 infections leads hospitalizations and 60 deaths annually to 2000 (USDA. 2008) . an estimated 73.000 E.

coli O157:H7 ƒ E. coli O157:H7 ƒ E. coli O157:H7 ƒ E. coli O157:H7 ƒ E. coli O157:H7 2010 ƒ E. coli O157:H7 Poultry 2009 ƒ E.2011 ƒ E. coli O157:H7 Lebanon Bologna Hazelnuts Bravo Farms Cheeses Romaine Lettuce from Beef from National Steak and Beef from Fairbank Farms Beef from JBS Swift Beef Company Prepackaged Cookie Dough . coli O145 : ƒ E.

2008) ƒ USDA and beef industry reported at least six other strains of STEC O26. O111. O45. coli O157:H7 (CDC..ƒ Non-O157 STEC strains are equally linked to several outbreaks and can be isolated in a similar frequency as E. coli O157:H7 (Bielaszewska et al. O103 that are equally as virulent as E. 2005) . O145.

coli O45 E. coli O145 CDC. coli O103 E.Occurrence Non-O157 Serotypes E. coli O26 E. coli O111 E. 2006 . coli O121 E.

ƒ 20-70 % of STEC infections throughout the world are due to non²O157 STEC (WHO. 1998) A four fold increase in incidence of non-O157 STEC has been reported from 2000-2006 (FoodNet. CDC) ƒ ƒ Non-O157 are underrepresented .

goat. pigs. where it is a benign commensal of the gastrointestinal tract (Galland et al..ƒ This pathogen is widely distributed within bovine populations. 2001) ƒ STEC isolates have been recovered from a variety of ruminant and non-ruminant: cattle. sheep. poultry. dogs and cats .

9% infection (Rangel et al..14% infection Environmental transmission .52% infection  41% ground beef  21% manufacturing process Person-to-person contact .There are 3 general modes of STEC transmission :  Foodborne transmission . 2005)   .

2005 .STEC Transmission Model Meat and Pet Animals Meat Cow Contact Water Cow Milk Water Human Human Manure Deer Fruits and vegetables Sulka.

com/tag/vre/ . coli (Maynard et al.ƒ The spread of antibiotic-resistant (ABR) bacteria is serious public health concerns (WHO.. 2004) http://benefitofmanukahoney. 2003) ƒ Antibiotics resistance for Ampicillin Tetracycline SulphamethoxazoleTrimethoprim. Chloramphenicol has increased among E.

1988) In more recent years.In 1980s. coli O157:H7 strains were resistant to an antibiotic (Ratnam et al. ABR was uncommon in E.. however.9% (5/174) E. coli O157:H7 ƒ 1% (2/200) of E. coli O157:H7 collected by CDC 1983 1985 were resistant to antibiotics (Bopp et al. 1987) ƒ 2.. the isolation of antibiotic resistance STEC has shown an increasing trend ƒ .

coli O157:H7 (Willshaw et al. showed similar resistance (SSxT) (Meng et al. (2005) reported >80% of 296 commensal E. coli isolates from animals were resistant to Tetracycline sulfamethoxazole and streptomycin ƒ . 1996) ƒ 70% E. 1998) Kang et al. coli O157:H7 isolated from ground beef.ƒ Streptomycin-sulfisoxazole-tetracycline (SSxT) resistance was dominant among E.

. O128.E. and O145 isolates. O111.2002) . ƒ Sulfamethoxazole . O103. coli O26.59% ƒ Streptomycin -59% ƒ Ampicillin -56% ƒ Tetracycline -56% ƒ Cephalothin -50% ƒ Trimethoprim-sulfamethoxazole -38% ƒ Chloramphenicol -34% (Schroeder et al.

coli O157:H7 in ground beef has been in place since 1996  Sorbitol-MacConkey agar supplemented with cefixime and tellurite (ct-SMAC) Chromocult Rainbow agar   .ƒ Zero-tolerance policy for the presence of E.

Specimen Colorless colony on SMAC agar Agglutination in O157 antiserum Pulsed field gel electrophoresis PFGE Patterns to PulseNet .

Stool Specimen GN broth Stx EIA Note: no SMAC plate. no PFGE. no colony. no PulseNet .

coli Long identification time 2-4 days Sorbitol MacConkey agar (SMAC) .    Presence of sorbitol fermenting E. coli O157:H7 No selective medium available Look like ´normalµ E.

IMS Most commercial kits target  O157 antigen  Shiga-toxin antigen ƒ ƒ .ƒ Enzyme immunoassays (EIA). were first licensed in1995 for being used in labs ELISA. Latex agglutination.

Commercial Stx EIAs  Premier EHEC  ProSpecT Shiga Toxin  Duopath Verotoxin GLISA  ImmunoCard STAT! EHEC  BioStar OIA SHIGATOX .

ƒ Stx EIAs cannot differentiate  between E. coli O157:H7 and other STEC serotypes  between Stx1 and Stx2 (more serious symptoms) False positive reactions are not uncommon Many lab tests for only Shiga toxin antigen ƒ ƒ .

PFGE Gold Standard Drawbacks ƒ Its can not differentiate non-O157 serotypes Complicated data analysis may raised reproducibility issues ƒ .

coli O157:H7 and STEC ƒ ƒ Samples are enriched in selective broth PCR for presence of Shiga toxin & virulence genes USDA-ARS. Nebraska .ƒ A number of PCR based methods are available for the detection of E. Clay Center.

O145. O45. Valadez et al.Recently a Multiplex Taqman Real. Detection limit 1-10 cfu/ml (Fratamico et al. O103. O111.Time PCR based assay was developed for the detection: O26... O121. 091.2011.2010) Drawback: They are very expensive .

. 2011) . O111.O145 and also O157 A cloth-based hybridization array system Detection limit: 10-60 cfu/ml The advantages of this assay: ƒ Polyester cloth is inexpensive and rapid ƒ It detects 6 different STEC strain with virulence gene (Blais et al. O103.Macroarray system for detection of Non:O157 strains O26.

Diarrhoea/Blood Samples 36 % samples Reference Labs 64% onsite STEC testing 89% Culture based 7% Stx EIA 4% Culture based + EIA No lab used PCR based methods for detection of STEC (Hoefer et al.. 2011) .

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 No selective medium for non-O157 STEC  Lack of any test for simultaneous identification STEC and detection of Virulence and Antibiotic Resistance genes  Lack of clear guidelines for testing. maybe more. STEC are a natural part of the animal microflora  Non-O157 STEC is probably just as prevalent. interpretation of results. than O157 STEC. and reporting .

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