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.

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Frequency of Detection ofEscherichia coli,Salmonellaspp., andCampylobacterspp. in the Faeces of Wild Rats (Rattusspp.) in Trinidad and Tobago

Veterinary Medicine International ◽

10.4061/2011/686923 ◽

2011 ◽

Vol 2011 ◽

pp. 1-7 ◽

Cited By ~ 21

Author(s):

Comfort Nkogwe ◽

Juliah Raletobana ◽

Alva Stewart-Johnson ◽

Sharianne Suepaul ◽

Abiodun Adesiyun

Keyword(s):

Antimicrobial Agents ◽

Diffusion Method ◽

Disc Diffusion ◽

Macconkey Agar ◽

Disc Diffusion Method ◽

O157 Strain ◽

E Coli ◽

Faecal Samples ◽

Wild Rats

The study was conducted to determine the frequency of isolation ofSalmonella,CampylobacterandE. coliO157 in the faecal samples of rats trapped across the regional corporations in Trinidad and to assess their resistance to antimicrobial agents. A total of 204 rats were trapped for the detection of selected bacteria. Standard methods were used to isolateSalmonella,CampylobacterandE. coliO157. Characterization ofE. coliwas done on sorbitol MacConkey agar to determine non-sorbitol fermentation, blood agar to determine haemolytic and mucoid colonies and by usingE. coliO157 antiserum to determine O157 strain. The disc diffusion method was used to determine resistance to nine antimicrobial agents. Of the 204 rats, 4 (2.0%), 7 (3.4%) and 171 (83.8%) were positive forSalmonellaspp.,Campylobacterspp. andE. coli, respectively. Of the 171 isolates ofE. colitested 0 (0.0%), 25 (14.6%) and 19 (11.1%) were haemolytic, mucoid and non-sorbitol fermenters, respectively. All isolates were negative for the O157 strain. The frequency of resistance to the 9 antimicrobial agents tested was 75% (3 of 4) forSalmonella, 85.7% (6 of 7) ofCampylobacterspp. and 36.3% (62 of 171) forE. coli(;χ2).

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Frequency of Escherichia coli O157:H7 in Turkish Cattle

Journal of Food Protection ◽

10.4315/0362-028x-65.10.1637 ◽

2002 ◽

Vol 65 (10) ◽

pp. 1637-1640 ◽

Cited By ~ 15

Author(s):

AYSUN YILMAZ ◽

HUSEYIN GUN ◽

HUSEYIN YILMAZ

Keyword(s):

Escherichia Coli ◽

Urease Activity ◽

Sampling Method ◽

Immunomagnetic Separation ◽

Systematic Sampling ◽

Escherichia Coli O157 ◽

Macconkey Agar ◽

E Coli ◽

Systematic Sampling Method ◽

Age And Sex

In this study, five abattoirs in Istanbul were visited between January 2000 and April 2001. During these visits, 330 cattle were selected by a systematic sampling method. Cattle were examined clinically and breed, age, and sex were recorded. Rectal swabs were taken immediately after slaughter. Immunomagnetic separation was performed, and sorbitol-negative colonies were selected on sorbitol MacConkey agar with cefixime and tellurite (CT-SMAC agar). These colonies were checked for 4-methylenebelliferyl-β-d-glucuronide, indol, rhamnose, and urease activity and motility. Serotypes of bacteria were determined by using antisera specific for Escherichia coli O157 and H7. All cattle selected were clinically healthy. Of 88 sorbitol-negative colonies selected on CT-SMAC agar, isolates from only 14 (4.2%) cattle reacted with anti-O157, and 13 of these isolates also reacted with anti-H7. E. coli O157:H7 was isolated from all breeds, but the numbers of isolates were largest for Holstein and Swiss Brown cows. E. coli O157:H7 was most frequently isolated from 2-year-old cattle. Similarly, it was most frequently isolated from male cattle. E. coli O157:H7 was isolated from cattle slaughtered in four of the five abattoirs studied.

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Verotoxin-producingEscherichia coliinfections in Sheffield: cattle as a possible source

Epidemiology and Infection ◽

10.1017/s0950268800030156 ◽

1989 ◽

Vol 102 (3) ◽

pp. 439-445 ◽

Cited By ~ 51

Author(s):

P. A. Chapman ◽

D. J. Wright ◽

P. Norman

Keyword(s):

Escherichia Coli ◽

Haemolytic Uraemic Syndrome ◽

Matched Control ◽

Bloody Diarrhoea ◽

E Coli ◽

Faecal Samples ◽

O 157 ◽

Age And Sex

SUMMARYDuring 1986 and 1987, faecal samples from patients with haemorrhagie colitis (HC) or haemolytic-uraemic syndrome (HUS) were examined for evidence of infection by verotoxin-producingEscherichia coli(VTEC). During the 2-year period VTEC infections were found in 31 (78%) of 40 patients initially presenting with HC, and in 5 (63%) of 8 patients initially presenting with HUS, VTEC were found in only 2 (0·9%) of 229 age and sex matched control patients with acute non-bloody diarrhoea. All but one VTEC belonged toE. coliserogroup O 157. During 1987 this serogroup was isolated from 2 (1%) of 207 samples of faeces taken from cattle arriving at a Sheffield abattoir, indicating a possible source of these infections for man. We are unaware of previous reports of isolation of this organism from cattle in England.

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Occurrence of Escherichia coli non-susceptible to quinolones in faecal samples from fluoroquinolone-treated, contact and control pigs of different ages from 24 Swiss pig farms

Porcine Health Management ◽

10.1186/s40813-021-00209-y ◽

2021 ◽

Vol 7 (1) ◽

Author(s):

Morena Amsler ◽

Katrin Zurfluh ◽

Sonja Hartnack ◽

Xaver Sidler ◽

Roger Stephan ◽

...

Keyword(s):

Escherichia Coli ◽

Bacterial Infections ◽

Farm Animals ◽

Resistant Bacteria ◽

Control Group ◽

Reduced Frequency ◽

Colony Forming Units ◽

Faecal Samples ◽

And Control

Abstract Background Despite their indispensability in human medicine, fluoroquinolones (FQ) are used for the treatment of bacterial infections in farm animals which increases the risk of transferring FQ-resistant bacteria into the environment and via the food chain to humans. The objectives of this observational study were to follow-up of the presence of quinolone non-susceptible Escherichia coli (QNSE) qualitatively and quantitatively in faecal samples of pigs at four time points (2 weeks old, 4 weeks old, 2 weeks post weaning and during fattening period). Moreover differences between groups of FQ-treated pigs, pigs with contact to treated pigs and control pigs were investigated. Additionally, quinolone and FQ resistance of Escherichia coli isolates of the faecal samples were investigated by determining minimum inhibitory concentrations (MICs). Results 40.9% of 621 fecal samples contained QNSE. Proportion of samples with detectable QNSE from treated and contact pigs did not differ significantly and were highest in piglets of 2 and 4 weeks of age. However, the proportions of samples with QNSE were significantly lowest in control pigs (7/90; 7.8%; CI = 3.5–14.7%) among all groups. Also, the number of colony-forming units was lowest in both weaners and fattening pigs of the control group compared to treated and contact groups. Following CLSI human breakpoints, in total, 50.4% out of 254 isolates in faecal samples were intermediate or resistant to ciprofloxacin. Conclusions QNSE were present in faeces of pigs independent of age or FQ background but significantly less were found in pigs from farms without FQ usage. Due to the long half-life of FQ, it is likely that only a prolonged absence of fluoroquinolone treatments in pig farming will lead to a reduced frequency of QNSE in the farm environment. Solutions need to be found to minimise the emergence and transfer of quinolone and FQ-resistant bacteria from treated pigs to contact pigs and to farms without FQ usage.

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Validation of selective agars for detection and quantification of Escherichia coli resistant to critically important antimicrobials

10.1101/2021.06.18.449085 ◽

2021 ◽

Author(s):

Zheng Z Lee ◽

Rebecca Abraham ◽

Mark O'Dea ◽

Ali Harb ◽

Kelly Hunt ◽

...

Keyword(s):

Escherichia Coli ◽

Linear Models ◽

Macconkey Agar ◽

Generalised Linear Models ◽

E Coli ◽

Designed Experiments ◽

Faecal Samples ◽

Pure Cultures ◽

Design Factors ◽

Detection And Quantification

Success in the global fight against antimicrobial resistance (AMR) is likely to improve if surveillance can be performed more rapidly, affordably and on a larger scale. An approach based on robotics and agars incorporated with antimicrobials has enormous potential to achieve this. However, there is a need to identify the combinations of selective agars and key antimicrobials yielding the most accurate counts of susceptible and resistant organisms. A series of designed experiments involving 1,202 plates identified the best candidate-combinations from six commercially available agars and five antimicrobials using 18 Escherichia coli strains as either pure cultures or inoculums within faeces. The effect of various design factors on colony counts were analysed in generalised linear models. Without antimicrobials, Brilliance™ E. coli (Brilliance) and CHROMagar™ ECC (CHROMagar) agars yielded 28.9% and 23.5% more colonies than MacConkey agar. The order of superiority of agars remained unchanged when faecal samples with and without spiking of resistant E. coli were inoculated onto agars with or without specific antimicrobials. When incorporating antimicrobials at varying concentrations, it was revealed that ampicillin, tetracycline and ciprofloxacin are suitable for incorporation into Brilliance and CHROMagar agars at all defined concentrations. Gentamicin was only suitable for incorporation at 8 and 16 µg/mL while ceftiofur was only suitable at 1 µg/mL. CHROMagar™ ESBL agar supported growth of a wider diversity of extended-spectrum cephalosporin-resistant E. coli. The findings demonstrate the potential for combining robotics with agars to deliver AMR surveillance on a vast scale with greater sensitivity of detection and strategic relevance.

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Isolation and characterization of enterohaemorrhagic Escherichia coli O157[ratio ]H7 from cattle in Belgium and Poland

Epidemiology and Infection ◽

10.1017/s0950268802007197 ◽

2002 ◽

Vol 129 (1) ◽

pp. 41-47 ◽

Cited By ~ 18

Author(s):

A. V. TUTENEL ◽

D. PIERARD ◽

J. URADZINSKI ◽

E. JOZWIK ◽

M. PASTUSZCZAK ◽

...

Keyword(s):

Phage Type ◽

Immunomagnetic Separation ◽

E Coli ◽

Slaughter Cattle ◽

Isolation And Characterization ◽

Enterohaemorrhagic Escherichia Coli ◽

Faecal Samples ◽

Peptone Water ◽

Ferment Lactose

EHEC O157 were isolated from faeces of Belgian and Polish beef slaughter cattle. In Belgium, 1281 faecal samples were analysed by immunomagnetic separation [IMS] after enrichment in buffered peptone water from June 1998 till July 1999. Eighty-one samples (6.3%) were positive for E. coli O157. Phage type 8 was most frequently found. Bulls between 1 and 2 years old, slaughtered in September and October were most frequently found positive. Atypical biochemical features were observed in some isolates: 22 (27%) isolates were urease positive and 1 (1.2%) isolate was unable to ferment lactose. In Poland, 551 faecal samples, taken from January 1999 till December 1999, were examined using exactly the same techniques. Four faecal samples (0.7%) were positive for O157 EHEC, yielding seven phage type 8 isolates. All positive samples were from cattle younger than 2 years. Positive samples occurred in August, September and October.

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Infection with verocytotoxin-producing Escherichia coli O157 during a visit to an inner city open farm

Epidemiology and Infection ◽

10.1017/s0950268800004775 ◽

2000 ◽

Vol 125 (3) ◽

pp. 531-536 ◽

Cited By ~ 47

Author(s):

P. A. CHAPMAN ◽

J. CORNELL ◽

C. GREEN

Keyword(s):

Escherichia Coli ◽

Inner City ◽

Magnetic Beads ◽

Profile Analysis ◽

Enrichment Culture ◽

Phage Type ◽

Immunomagnetic Separation ◽

E Coli ◽

Zoonotic Infections ◽

Faecal Samples

Two cases of Escherichia coli O157 infection occurred in children after visiting an inner city open farm. Subsequently faecal samples collected from animal pens and samples of composted mixed animal manure and vegetable waste were examined for E. coli O157 by enrichment culture, immunomagnetic separation and culture of magnetic beads to cefixime tellurite sorbitol MacConkey agar. Strains of E. coli O157 were characterized by hybridization with DNA probes for VT1, VT2 and eaeA, plasmid profile analysis, phage typing and pulsed field gel electrophoresis (PFGE). Verocytotoxin-producing E. coli O157 strains were isolated from faecal samples from a cow, a horse, 3 breeds of pigs, 2 breeds of sheep and 2 breeds of goats and from 2 samples of compost which had been processed for 3 months. All strains were phage type 21, hybridized with probes for VT2 and eaeA but not with one for VT1, harboured 92 and 2 kb plasmids and gave indistinguishable banding patterns with PFGE. Although only two culture-confirmed cases of infection had been identified, the farm had over 100 000 visitors per year and so it was closed as a precaution both to allow a thorough investigation and to prevent further cases. The investigation identified many factors which may have contributed to transmission of E. coli O157 infection. Most of these were readily resolved by appropriate corrective measures and as there were no further cases associated with the farm during the ensuing 4 weeks it then re-opened. These cases highlight the risk, especially to young children, of acquiring zoonotic infections during visits to open farms and emphasize the need for adequate guidance and supervision before and during such visits.

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Detection and characterisation of extended-spectrum and plasmid-mediated AmpC β-lactamase produced by Escherichia coli isolates found at poultry farms in Bosnia and Herzegovina

Archives of Industrial Hygiene and Toxicology ◽

10.2478/aiht-2021-72-3560 ◽

2021 ◽

Vol 72 (4) ◽

pp. 305-314

Author(s):

Majda Fetahagić ◽

Amir Ibrahimagić ◽

Selma Uzunović ◽

Nataša Beader ◽

Vesna Elveđi-Gašparović ◽

...

Keyword(s):

Escherichia Coli ◽

Bosnia And Herzegovina ◽

Farm Animals ◽

E Coli ◽

Extended Spectrum ◽

Faecal Samples ◽

Poultry Farms ◽

Genes Encoding ◽

Polymerase Chain ◽

Sequence Types

Abstract Extended-spectrum β-lactamases (ESBLs) hydrolyse extended-spectrum cephalosporins (ESC) and aztreonam. As ESBL-producing organisms have been identified in food producing animals, the aim of our study was to detect and analyse such Escherichia coli isolates from poultry. Antibiotic susceptibility of the isolates was determined with disk-diffusion and broth microdilution methods. ESBLs were detected with the double-disk synergy and inhibitor-based test with clavulanic acid. The transferability of cefotaxime resistance was determined with conjugation experiments, and genes encoding ESBLs, plasmid-mediated AmpC β-lactamases, and quinolone resistance determinants identified by polymerase chain reaction. The study included 108 faecal samples (cloacal swabs) from 25 different poultry farms in the Zenica-Doboj Canton, Bosnia and Herzegovina. Of these, 75 (69.4 %) were positive for E. coli, of which 27 were resistant to cefotaxime, amoxicillin, cefazoline, and cefriaxone, and susceptible to imipenem, meropenem, ertapenem, and amikacin. All 27 cefotaxime-resistant isolates were positive in double-disk synergy and combined disk tests. Eighteen isolates transferred cefotaxime resistance to E. coli recipient. Twenty-one isolates were positive for the bla CTX-M-1 cluster genes and seven for bla CTX-M-15. Fourteen were positive for the bla TEM genes. The most frequent plasmid incompatibility group was IncFIB, whereas IncFIA and Inc HI1 were present in only a few isolates. Two different sequence types (STs) were identified: ST117 and ST155. The emergence of ESBL-producing E. coli in farm animals presents a public health threat, as they can colonise the intestine and cause infections in humans.

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Occurrence of shigatoxinogenic Escherichia coli O157 in Norwegian cattle herds

Epidemiology and Infection ◽

10.1017/s0950268897008364 ◽

1998 ◽

Vol 120 (1) ◽

pp. 21-28 ◽

Cited By ~ 29

Author(s):

L. VOLD ◽

B. KLUNGSETH JOHANSEN ◽

H. KRUSE ◽

E. SKJERVE ◽

Y. WASTESON

Keyword(s):

Escherichia Coli ◽

Vero Cell ◽

Immunomagnetic Separation ◽

The Other ◽

Virulence Plasmid ◽

Escherichia Coli O157 ◽

E Coli ◽

Faecal Samples ◽

Cattle Herds ◽

Encoding Genes

To investigate if there is a reservoir of Escherichia coli O157 in Norwegian cattle, faecal samples from 197 cattle herds were screened for E. coli O157 by the use of immunomagnetic separation (IMS) and PCR during the 1995 grazing season. Six E. coli O157[ratio ]H-isolates were detected in two herds, one isolate in one and five in the other. The isolates carried the stx1, stx2, and eae genes, and a 90 MDa virulence plasmid. They were toxinogenic in a Vero cell assay. From 57 other herds, 137 faecal samples were positive for stx1 and/or stx2 genes detected by PCR run directly on IMS-isolated material. Among these samples, stx2 were the most widely distributed toxin encoding genes. No difference was found among milking cows and heifers in the rate of stx1 and/or stx2 in positive samples.

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The use of sorbitol-MacConkey agar in conjunction with a specific antiserum for the detection of Vero cytotoxin-producing strains ofEscherichia coliO 157

Epidemiology and Infection ◽

10.1017/s0950268800054261 ◽

1988 ◽

Vol 101 (2) ◽

pp. 327-335 ◽

Cited By ~ 11

Author(s):

H. Kleanthous ◽

N. K. Fry ◽

H. R. Smith ◽

R. J. Gross ◽

B. Rowe

Keyword(s):

Escherichia Coli ◽

Haemolytic Uraemic Syndrome ◽

Screening Method ◽

Bloody Diarrhoea ◽

Macconkey Agar ◽

Haemorrhagic Colitis ◽

Faecal Samples ◽

Genes Encoding ◽

Homologous Antiserum ◽

O 157

SUMMARYUsing DNA probes specific for the genes encoding Vero cytotoxins 1 and 2 in hybridization experiments on faecal samples. Vero cytotoxin-producingEscherichia coli(VTEC) of serogroup 0 157 were detected in 21 of 63 cases of haemorrhagic colitis, 9 of 31 cases of non-bloody diarrhoea and 14 of 68 cases of haemolytic uraemic syndrome. Compared with these results sorbitol-MacConkey agar in conjunction with a specific 0 157 antiserum gave a sensitivity of 62% in haemorrhagic colitis, 56% in non-bloody diarrhoea and 57% in haemolytic uraemic syndrome.The specificity of this method was 100% in all three groups. This demonstrates that sorbitol-MacConkey agar is a useful screening method for the detection of VTEC of serogroup O 157 when used in conjunction with a specific homologous antiserum. However, this method does not detect VTEC belonging to other serogroups and such strains were found, particularly in cases of haemolytic uraemic syndrome.

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