OIE Reference Laboratory Reports

Activities in 2011

Name of disease (or topic) for Escherichia coli

which you are a designated OIE
Reference Laboratory:

Address of laboratory: Reference Laboratory for Escherichia coli (EcL)

Faculté de médecine vétérinaire
Université de Montréal
3200 Sicotte St.
Saint-Hyacinthe (Quebec)
CANADA J2S 2M2

Tel.: + 1-450.773.8521

Fax: +1-450.778.8108

e-mail address: john.morris.fairbrother@umontreal.ca

websites: www.ecl-lab.ca
www.apzec.ca

Name (including Title and John M. FAIRBROTHER

Position) of Head of Laboratory Professor
(Responsible Official):

Name(including Title and John M. FAIRBROTHER

Position) of OIE Reference Professor
Expert:

Name (including Title and

Position) of writer of this report
(if different from above):

Annual reports of OIE Reference Centres, 2011 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 1992

PCR Virotyping of E. coli isolates 3–6 virulence factor genes 1882

Gene probe Virotyping of E. coli isolates 20 virulence factor genes 774

Slide agglutination Serotyping of E. coli isolates O serotyping 152

PCR Human pathogenic E. coli isolates

O157:H7 672
in animal samples

2. Production and distribution of diagnostic reagents

Amount supplied nationally Amount supplied to other

Type of reagent
(including for own use) countries

F4 (K88) antiserum for slide

70
agglutination (1 ml vial)

F5 (K99) antiserum for slide

92
agglutination (1 ml vial)

E. coli reference strains 10 2 (Costa Rica); 1 (USA)

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

a) Establishment and maintenance of a network with other OIE Reference Laboratories

designated for the same pathogen or disease and organisation of regular inter-laboratory
proficiency testing to ensure comparability of results

As part of its mandate as an OIE reference laboratory, the EcL has developed the Global “Animal pathogenic
and zoonotic Escherichia coli” (APZEC) Database, in order to centralize worldwide surveillance data on
APZEC. The surveillance programme integrates virotype and antimicrobial resistance data on APZEC. E.
coli isolates are characterised for virotype, as determined by PCR and gene probe testing for the genes of
over 20 virulence factors, and for O serotype. We are currently developing a ring assay to harmonise the
procedure and the reagents used in the virotyping. Laboratories undertaking this evaluation will then submit
data directly to the data base housed in our laboratory.

b) Organisation of inter-laboratory proficiency testing with laboratories other than OIE

Reference Laboratories for the same pathogens and diseases to ensure equivalence of
results

2 Annual reports of OIE Reference Centres, 2011

 Escherichia coli

We participated in the 3rd 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. This programme focussed on
the subtyping of VT-encoding genes (vtx) by conventional PCR amplification of the different gene subtypes.

4. Preparation and supply of international reference standards for diagnostic tests or vaccines

We prepare and provide more than 200 reference E. coli strains and accompanying data sheets for virotyping, O
serotyping, and antimicrobial resistance testing of animal pathogenic and zoonotic E. coli by PCR, colony
hybridisation, agglutination, disk diffusion, 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 2012, 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 3).

We have now included primers for the detection of important O serotypes and antimicrobial resistance genes such
as those for fluoroquinolones in our PCR. The PCR will, to a large degree, replace the standard agglutination test
for O serotyping. The PCR for detection of antimicrobial resistance genes will be used mostly for surveillance
purposes. These PCR tests will be validated in 2012.

We have optimised an approach for the transport of primary cultures on MacConkey agar of potentially pathogenic
E. coli isolated from faecal samples taken on farm in OIE member countries. These are transported to our
laboratory for rapid detection of E. coli pathotypes and subsequent identification of the virotype and determination
of the presence of antimicrobial resistance genes in individual selected isolates.

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. The
Global APZEC Database will be available on the website for the EcL from early 2012. 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.

Specific scientific projects in 2011 include:

Time
Purpose Participants Outputs Follow up plans
frame

Investigating E. coli virotypes University of 2009- Characterisation of 138 Analysis and

and antibiotic resistance New South ongoing isolates by PCR and publication of
patterns in wild populations of Wales, Sydney, antimicrobial resistance results
Southern elephant seals, Australia by disk diffusion

Annual reports of OIE Reference Centres, 2011 3

 Escherichia coli

Antarctica

Time
Purpose Participants Outputs Follow up plans
frame

Identification of STEC
Prevalence and characterization University of Sao and EPEC strains,
2009- Publication in
of potentially pathogenic E. coli Paulo State characterisation by
ongoing preparation
in healthy sheep in Brazil (UNESP), Brazil PFGE and sequencing
of virulence genes
Characterisation of 167
isolates by PCR,
Prevalence and characterization
University of Sao PFGE, disk diffusion Analysis and
of potentially pathogenic E. coli
Paulo State 2010 technique for publication of
in unpasteurised cheese in
(UNESP), Brazil antimicrobial results
Brazil
resistance, serotyping,
phylogeny by PCR
Demonstration of
Survey of food
Virotype and antimicrobial National Institute unique virotypes and
born potentially
resistance of E. coli isolates of Veterinary 2008- antimicrobial resistance
zoonotic E. coli
from pigs and chickens on farm Research (NIVR), ongoing patterns in Vietnam
in pigs and
and at slaughter in Vietnam Vietnam (see section 12 for
chickens
publications)
National
Virotype and antimicrobial Veterinary
Analysis and
resistance of E. coli isolates Research and 2009- O serotyping of 150
publication of
from pigs with diarrhoea in Quarantine ongoing isolates
results
South Korea Service(NVRQS),
South Korea
École Inter-États Continued
Virotype and antimicrobial des Sciences et Identification of characterisation
resistance of E. coli isolates Médecine 2011- potentially pathogenic of isolates,
from chickens on farm and at Vétérinaires ongoing E. coli and their analysis,
slaughter in Sénégal (EISMV), antimicrobial resistance publication of
Sénégal results

7. Maintenance of a system of quality assurance, biosafety and biosecurity relevant to the

pathogen and the disease concerned

The Diagnostic Service of the Faculty of Veterinary Medicine, of which our laboratory is a part, is currently
undergoing accreditation by the American Association of Veterinary Laboratory Diagnosticians (AAVLD). The
purpose of AAVLD accreditation program is to accredit public veterinary diagnostic laboratories in North
America relative to technical and operational competence compatible with appropriate standards, and to provide
an administrative assessment.

Our laboratory is a Biosafety Level 2 Laboratory, permitting us to work with pathogenic E. coli, including
VTEC/STEC. Our laboratory is certified by the Public Health Agency of Canada and the Canadian Food
Inspection Agency to import E. coli which are human and/or animal pathogens into Canada.

8. Provision of consultant expertise to OIE or to OIE Member Countries

Dr. Fairbrother carried out a review of the scientific literature on the control of VTEC in food producing animals
for presentation to the OIE Animal Production Food Safety Working Group. This report will provide the basis for
a decision by the OIE on future priorities for standards development in relation to the on-farm control of food
borne pathogens.

4 Annual reports of OIE Reference Centres, 2011

 Escherichia coli

Dr. Fairbrother carried out a one week mission at the École Inter-États des Sciences et Médecine Vétérinaires
(Sénégal), to provide advice on setting up a project on the study of virotype and antimicrobial resistance of E. coli
isolates from chickens on farm and at slaughter in Sénégal.

He carried out a mission to Brazil to provide advice to swine veterinarians on alternative approaches to the use of
antimicrobial agents for the control E. coli disease in pigs.

9. Provision of scientific and technical training to personnel from other OIE Member Countries

The following personnel were trained in my laboratory on procedures for detection and identification of
pathogenic E. coli and determination of antimicrobial resistance.

Personnel OIE Member Country Time of stay

MSc student Cameroun 2 years (ongoing)
MSc student Sénégal/Tchad 3 months
MSc student Vietnam 2 years (ongoing)
PhD student Korea 3 days

10. Provision of diagnostic testing facilities to other OIE Member Countries

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.

11. Organisation of international scientific meetings on behalf of OIE or other international bodies

No activities.

12. 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 and chickens on
farm and at slaughter, in different geographical regions and production systems. Dr. Fairbrother carried out a one
week mission to Hanoi, Vietnam, meeting the different Institutes of the Ministry Of Agriculture and Rural
Development to explore the possibility of setting up a surveillance of pathogenic E. coli and antimicrobial
resistance in production animals on farm.

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.

13. 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

Jahanbakhsh S., P.K. Kabore, P. Fravalo, A. Letellier, J.M. Fairbrother. Effect of dietary Clinoptilolite on fecal
shedding of blaCMY-2 –positive E. coli in healthy pigs. Antimicrobial Stewardship in Canadian Agriculture and
Veterinary Medicine Conference, Oct. 30-Nov. 2, 2011.

Annual reports of OIE Reference Centres, 2011 5

 Escherichia coli

Fairbrother, J.M., C. Desautels, H.R. Kohan-Ghadr, E. Nadeau. Global animal pathogenic and zoonotic E. coli
(APZEC) database. Antimicrobial Stewardship in Canadian Agriculture and Veterinary Medicine Conference, Oct.
30-Nov. 2 2011.

Byun, J., B.Y. Jung, H.Y. Kim, J.M. Fairbrother, W.K. Lee. Real-time PCR for differentiation of F4 variants
(F4ab, F4ac, and F4ad) of enterotoxigenic E. coli isolated from diarrheic piglets. 115th Annual Meeting United
States Animal Health Association and 54th Annual Conference American Association of Veterinary Laboratory
Diagnosticians, Buffalo, New York, Sept. 29-Oct. 5 2011.

Lessard, M., J. P. Brousseau, J. F. Daudelin, F. Beaudoin, É. Nadeau, G. Talbot, D. Roy, J. M. Fairbrother.

Microbial diversity and intestinal immunity of weanling piglets modulated by probiotics. 15th International
Congress of Mucosal Immunology (ICMI 2011), Paris, France, July 5-9 2011.

Fairbrother, J.M. Update on worldwide E. coli disease: Incidence, impact and control. 6th Suinter 2011 (Sympósio
Internacional de Produçăo Suína), Iguazu Falls, Brazil, June 27, 2011.

Fairbrother, J.M., C. Desautels, É. Nadeau. Prevalence of pathogenic Escherichia coli in production animals in
Québec, Canada, between 2008 and 2010. 10th Annual meeting of the Canadian Animal Health Laboratorians
Network (CAHLN), New Technologies - New Horizons, June 5-8, 2011.

Fairbrother, J.M. Nouvelles tendances dans les Escherichia coli pathogènes chez le porc, le bovin et la volaille.
5ème Colloque International Francophone de Microbiologie Animale, Marrakech, Maroc, April 5 2011.

 Scientific publications in peer-reviewed journals

Smith, M.G., D. Jordan, T.A. Chapman, J.J-C Chin, M.D. Barton, T.N. Do, V.A. Fahy, J.M. Fairbrother, D.J.
Trott. 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.

Daudelin, J.F., M. Lessard, F. Beaudoin, É. Nadeau, N. Bissonnette, Y. Boutin, J.P. Brousseau, K. Lauzon, J.M.
Fairbrother. 2011. Administration of probiotics influences F4 positive enterotoxigenic Escherichia coli attachment
and intestinal cytokine expression in weaned pigs. Veterinary Research 42:69 pp. 1-11.

 Other communications

Fairbrother, J.M. 2011. Prevención de diarreas por Escherichia coli en lechones (Prevention of E. coli diarrhoea in
pigs). Suis 81: 14-22.

Expert Panel on Approaches to Animal Health Risk Assessment. 2011. Healthy animals, healthy Canada. Council
of Canadian Academies. 267 p.

 Internet websites

www.ecl-lab.ca/en
www.apzec.ca

_______________

6 Annual reports of OIE Reference Centres, 2011