OIE Reference Laboratory Reports
Activities in 2010
Name of disease (or topic) for
which you are a designated OIE
Reference Laboratory:
Address of laboratory:
Escherichia coli
Reference Laboratory for Escherichia coli (EcL)
Faculté de médecine vétérinaire
Université de Montréal
3200 Sicotte St.
Saint-Hyacinthe (Quebec)
Canada J2S 2M2
Tel.:
(+1-450) 773.85.21
Fax:
(+1-450) 778.81.08
e-mail address:
john.morris.fairbrother@umontreal.ca
www.ecl-lab.ca
websites:
www.apzec.ca
Name of Head of Laboratory
(Responsible Official):
John M. FAIRBROTHER
Name of OIE Reference Expert:
John M. FAIRBROTHER
Name of writer of this report
(if different from above):
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
1
Escherichia coli
Part I: Summary of general activities related to the disease
1.
Test(s) in use/or available for the specified disease/topic at your laboratory
Test
For
Specificity
Total
PCR
E. coli pathotypes in samples
3-6 virulence factor genes
1412
PCR
Virotyping of E. coli isolates
3-6 virulence factor genes
1433
Gene probe
Virotyping of E. coli isolates
20 virulence factor genes
584
Slide agglutination
Serotyping of E. coli isolates
O serotyping
109
PCR
Human pathogenic E. coli isolates
in animal samples
O157:H7
298
Samples were mostly faeces or intestinal contents. E. coli pathotypes present in samples were identified by PCR
testing for the genes of virulence factors STa, STb, LT, Eae, F4, Stx1, and/or Stx2. Isolates were tested by PCR for
virotype based on the presence of the genes for virulence factors STa, STb, LT, Eae, F4, Stx1, and/or Stx2.
Isolates were tested by gene probe for virotype based on the presence of the genes for up to 20 virulence factors,
including STa, STb, LT, Eae, F4, Stx1, and/or Stx2. Isolates were tested for serotype by slide agglutination using
antisera for up to 86 O serotypes.
2.
Production and distribution of diagnostic reagents
Type of reagent
Amount supplied nationally
(including for own use)
Pool 1N OK antiserum for slide
agglutination (1 ml vial)
10
Pool 2N OK antiserum for slide
agglutination (1 ml vial)
2
Individual O antisera for slide
agglutination (1 ml vial)
70
Individual OK antisera for slide
agglutination (1 ml vial)
3
F4 (K88) antiserum for slide
agglutination (1 ml vial)
64
F5 (K99) antiserum for slide
agglutination (1 ml vial)
58
E.coli reference strains
78
Amount supplied to other
countries
6 (France)
68 (South Korea)
52 (USA)
Pool 1N comprises 9 OK antisera usually associated with neonatal diarrhoea in pigs and calves. Pool 2N
comprises 10 antisera usually associated with postweaning diarrhoea and oedema disease in pigs. F4 (K88)
fimbrial adhesin is expressed by many E. coli causing diarrhoea in pigs. F5 (K99), F6 (987P) and F41 fimbrial
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Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
Escherichia coli
adhesins is expressed by many E. coli causing diarrhoea in pigs and calves.
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
3
Escherichia coli
Part II: Activities specifically related to the mandate
of OIE Reference Laboratories
3.
International harmonisation and standardisation of methods for diagnostic testing or the
production and testing of vaccines
We participated in the 2nd round of the External Quality Assurance (EQA) ring test for verocytotoxin-producing
E. coli (VTEC) typing organized by the International Escherichia and Klebsiella Centre (WHO) at Statens Serum
Institut (SSI), Denmark, for the European Centre for Disease Prevention and Control (ECDC). This laboratory
organizes annual EQA rounds for the national reference laboratories in the EU and European Economic Area
(EEA) countries on serotyping and virulence typing for VTEC. We are currently participating in the Third ECDC
EQA on Verocytotoxin encoding genes subtyping.
4.
Preparation and supply of international reference standards for diagnostic tests or vaccines
We prepare and supply more than 70 reference E. coli strains and accompanying data sheets for virotyping and O
serotyping of animal pathogenic and zoonotic E. coli by PCR, colony hybridisation, agglutination, etc.
5.
Research and development of new procedures for diagnosis and control
A PCR for the rapid detection of E. coli pathotypes in faecal and other tissue samples from animals and
subsequent identification of the virotype and determination of the presence of antimicrobial resistance genes has
been set up. This procedure is already used in many laboratories. We are standardizing this test and the protocol is
available on our website (http://www.apzec.ca/en/APZEC/Protocols/APZEC_PCR_en.aspx ). In 2011, we will
carry out interlaboratory test evaluations to harmonise the procedure and the reagents used. Laboratories
undertaking this evaluation will then submit data directly to the data base set up in our laboratory (see Section 6).
A procedure using non-radioactive DIG colony hybridisation has been developed for the identification of the
virotype and determination of the presence of antimicrobial resistance genes in isolates from PCR-positive
samples. We are currently validating this procedure. This technique will be particularly valuable for the
identification of pathogenic and antimicrobial resistant E. coli in environmental samples, food products, and
healthy animal carriers of zoonotic E. coli.
6.
Collection, analysis and dissemination of epizootiological data relevant to international disease
control
As part of its mandate as an OIE reference laboratory, the EcL has developed the global “Animal pathogenic and
zoonotic Escherichia coli” (APZEC) Project. The mission of this project is to become an international reference
centre on E. coli, centralizing worldwide surveillance data on APZEC. The global APZEC project has 3 main
themes: establishment of a global, integrated surveillance system for APZEC, development of applied research
projects in collaboration with the international scientific community, and education on APZEC. Through this
project, the EcL is developing partnerships with laboratories in different countries. The surveillance programme
integrates virotype and antimicrobial resistance data on APZEC. A surveillance data base has been developed at
the EcL and will permit mapping of the geographical distribution of virotypes in different animal species. This
data will be reported on the website for the EcL from early 2011. Representative isolates are sent to the EcL for
validation of data and further in-depth characterization to monitor the emergence of new pathogenic strains. We
are building up a reference collection of APZEC, which are characterised for virotype, as determined by PCR and
gene probe testing for the genes of over 20 virulence factors, and for O serotype. Strains are also characterised for
the pulse-field gel electrophoresis profile.
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Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
Escherichia coli
Specific scientific projects in 2010 include:
Purpose
Investigating E. coli virotypes
and antibiotic resistance
patterns in wild populations of
Southern elephant seals,
Antarctica
Prevalence and characterization
of potentially pathogenic E. coli
in healthy sheep in Brazil
Prevalence and characterization
of potentially pathogenic E. coli
in unpasteurised cheese in
Brazil
Virotypes and antimicrobial
resistance of E. coli isolates
from pigs with diarrhea in Ivory
Coast
7.
Participants
Time
frame
Outputs
Follow up plans
University of
New South
Wales, Sydney,
Australia
2009ongoing
Characterisation of 138
isolates by PCR and
antimicrobial resistance
by disk diffusion
Analysis and
publication of
results
2009ongoing
Identification of STEC
and EPEC strains,
characterisation by
PFGE and sequencing
of virulence genes
Publication in
preparation
2010
Characterisation of 167
isolates by PCR,
PFGE, disk diffusion
technique for
antimicrobial
resistance, serotyping,
phylogeny by PCR
Analysis and
publication of
results
2009ongoing
Characterisation of 236
isolates by PCR and
non-radioactive DIG
colony hybridisation
for presence of
virulence and
antimicrobial resistance
genes, antimicrobial
resistance by disk
diffusion
Analysis and
publication of
results
Survey of
potentially
zoonotic E. coli
in cattle, pigs,
other production
species
Analysis and
publication of
results
University of Sao
Paulo State,
Brazil
University of Sao
Paulo State,
Brazil
University of
Abobo-Adjamé,
Ivory Coast
Virotype and antimicrobial
resistance of E. coli isolates
from pigs with diarrhea in
Vietnam
National Institute
of Veterinary
Research (NIVR),
Vietnam
2008ongoing
Demonstration of
unique virotypes and
antimicrobial resistance
patterns in Vietnam
(see section 12 for
publications)
Virotype and antimicrobial
resistance of E. coli isolates
from pigs with diarrhea in
South Korea
National
Veterinary
Research and
Quarantine
Service(NVRQS),
South Korea
2009ongoing
O serotyping of 100
isolates
Provision of consultant expertise to OIE or to OIE Members
Dr. Fairbrother participated in a Verocytotoxin/Shiga toxin-producing E. coli (VTEC/STEC) Expert Group for
DISCONTOOLS. The DISCONTOOLS project is a joint initiative of industry and a wide range of stakeholders
including the research community, regulators, users and others. It is actively encouraged and funded by the
European Commission services and will provide a mechanism for focusing and prioritising research that ultimately
delivers new and improved vaccines, pharmaceuticals and diagnostic tests.
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
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Escherichia coli
8.
Provision of scientific and technical training to personnel from other OIE Members
The following personnel were trained in my laboratory on procedures for detection and identification of
pathogenic E. coli.
9.
Personnel
OIE Member
Time of stay
PhD student
Australia
4 weeks
PhD student
Brazil
4 months
DVM student
Brazil
4 months
PhD student
Ivory Coast
6 months
Researcher
Vietnam
3 weeks
MSc student
Vietnam
4 weeks (ongoing)
Provision of diagnostic testing facilities to other OIE Members
Virotyping and serotyping was carried out on E. coli isolates from pigs and chickens in Vietnam, France, and
South Korea. To date, E. coli is not a reportable disease.
10. Organisation of international scientific meetings on behalf of OIE or other international bodies
No activities.
11. Participation in international scientific collaborative studies
Continued participation in a collaborative study with the National Institute of Veterinary Research (NIVR), in
Vietnam, to compare trends in virotype and antimicrobial resistance of E. coli isolates from pigs with diarrhoea, in
different geographical regions and production systems.
As a part of the global APZEC Project (see section 6), initiation of participation in collaborative studies to
determine the prevalence of pathogenic E. coli in animals with diarrhoea and to identify animal reservoirs for
zoonotic E. coli with 1) Korean National Veterinary Research and Quarantine Service; 2) University of NSW,
Australia; 3) University of Sao Paulo, Brazil; 4) Prevtec do Brazil; 5) University of Abobo-Adjamé, Ivory Coast;
and 6) École Inter-États des Sciences et de Médecine Vétérinaires (EISMV), Sénégal.
12. Publication and dissemination of information relevant to the work of OIE (including list of
scientific publications, internet publishing activities, presentations at international conferences)

Presentations at international conferences and meetings
Nadeau E, Tremblay D, Fairbrother JM. Vaccination with Coliprotec® vaccine for the prevention of post-weaning
diarrhea associated with F4(K88)-positive enterotoxigenic Escherichia coli (ETEC). Proceedings of the 21st
International Pig Veterinary society congress, Vancouver, Canada. July 18-21 2010. International Pig Veterinary
Society 2010. p. 463.
Do TN, Trott DJ, Nadeau E, Desautels C, Fairbrother JM. Comparison of the pathotypes and virotypes of
pathogenic Escherichia coli in diseased pigs in Vietnam and Quebec, Canada. Proceedings of the 21st IPVS
Congress, Vancouver, Canada. July 18 to 21, 2010. International Pig Veterinary Society 2010. p. 766.
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Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
Escherichia coli
Do TN, Trott DJ, Nadeau E, Desautels C, Fairbrother JM. Comparison of the antimicrobial resistance of
pathogenic Escherichia coli in diseased pigs in Vietnam and Quebec, Canada. Proceedings of the 21st IPVS
Congress, Vancouver, Canada. July 18 to 21, 2010. International Pig Veterinary Society 2010. p. 774.
Kaboré PK, Letellier A, Fravalo P, Lahaye L, Fairbrother JM. Prevalence and association of virulence and
antimicrobial resistance genes of avian pathogenic E. coli from the intestinal microflora of healthy chickens. 2nd
ASM Conference on Antimicrobial Resistance in Zoonotic Bacteria and Foodborne Pathogens in Animals,
Humans and the Environment, Toronto, Canada, June 8-11, 2010, p. 33.
Kaboré PK, Letellier A, Fravalo P, Lahaye L, Fairbrother JM. Prevalence and association of virulence and
antimicrobial resistance genes of avian pathogenic E. coli from the intestinal microflora of healthy chickens. 2nd
ASM Conference on Antimicrobial Resistance in Zoonotic Bacteria and Foodborne Pathogens in Animals,
Humans and the Environment, Toronto, Canada, June 8-11, 2010, p.113.
Jahanbakhsh S, Desautels C, Desrosiers A, l’Homme Y, Fairbrother JM. Antimicrobial resistance in potentially
pathogenic Escherichia coli isolated from oysters from prohibited growing areas in Canada. 2nd ASM Conference
on Antimicrobial Resistance in Zoonotic Bacteria and Foodborne Pathogens in Animals, Humans and the
Environment, Toronto, Canada, June 8-11, 2010, p. 80.

Scientific publications in peer-reviewed journals
Smith MG, Jordan D, Chapman TA, Chin JJ-C, Barton MD, Do TN, Fahy VA, Fairbrother JM, Trott DJ. 2010.
Antimicrobial resistance and virulence gene profiles in multi-drug resistant enterotoxigenic Escherichia coli
isolated from pigs with post-weaning diarrhoea. Veterinary Microbiology. In press.
Lefebvre B, Diarra MS, Fairbrother JM, Nadeau E, Dubois MJr, Malouin F. 2010. Intestinal Mucosa Adherence
and Cytotoxicity of A Sorbitol-Fermenting, Shiga Toxin-Negative Escherichia coli O157 Isolate With an Atypical
Type III Secretion System. Foodborne Pathogens and Disease 7: 985-990.

Other communications
Fairbrother JM and Gyles CL. Escherichia coli. In Diseases of Swine. Straw BE, Zimmerman JJ, D’Allaire S,
Taylor DJ, editors. Iowa State University Press. Ames, Iowa, USA. 10th edition. In press.
Fairbrother JM and Nadeau E. 2010. Colibacillosis. In Infectious and Parasitic Diseases of Livestock. Lefèvre PC,
Blancou J, Chermette R, Uilenberg G, editors. Lavoisier. Vol. 2, Chapter 74: 917-945.
Gyles CL and Fairbrother JM. 2010. Escherichia coli. In Pathogenesis of Bacterial Infections in Animals. Gyles
CL, Prescott JF, Songer JG, Thoen CO, editors. Wiley-Blackwell. Ames, Iowa, USA. 4th edition. Chapter 15, pp.
267-308.

Internet websites
www.ecl-lab.ca/en
www.apzec.ca
13. Inscription of diagnostic kits on the OIE Register
i)
Did you participate in expert panels for the validation of candidate kits for inscription on the
OIE Register? If yes, for which kits?
No
ii)
Did you submit to the OIE candidate kits for inscription on the OIE Register? If yes, for
which kits?
No
_______________
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
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