Escherichia coli
OIE Reference Laboratory Reports
Activities in 2011
Name of disease (or topic) for which you are a designated OIE Reference Laboratory: / Escherichia coliAddress 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: /
websites: / www.ecl-lab.ca
www.apzec.ca
Name (including Title and Position) of Head of Laboratory (Responsible Official): / John M. FAIRBROTHER
Professor
Name(including Title and Position) of OIE Reference Expert: / John M. FAIRBROTHER
Professor
Name (including Title and Position) of writer of this report
(if different from above):
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 / TotalPCR / 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 in animal samples / O157:H7 / 672
2. Production and distribution of diagnostic reagents
Type of reagent / Amount supplied nationally(including for own use) / Amount supplied to other countries
F4 (K88) antiserum for slide agglutination (1ml vial) / 70
F5 (K99) antiserum for slide agglutination (1ml vial) / 92
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
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:
Purpose / Participants / Time frame / Outputs / Follow up plansInvestigating E. coli virotypes and antibiotic resistance patterns in wild populations of Southern elephant seals, Antarctica / University of New South Wales, Sydney, Australia / 2009- ongoing / Characterisation of 138 isolates by PCR and antimicrobial resistance by disk diffusion / Analysis and publication of results
Purpose / Participants / Time frame / Outputs / Follow up plans
Prevalence and characterization of potentially pathogenic E. coli in healthy sheep in Brazil / University of Sao Paulo State (UNESP), Brazil / 2009-ongoing / Identification of STEC and EPEC strains, characterisation by PFGE and sequencing of virulence genes / Publication in preparation
Prevalence and characterization of potentially pathogenic E. coli in unpasteurised cheese in Brazil / University of Sao Paulo State (UNESP), Brazil / 2010 / Characterisation of 167 isolates by PCR, PFGE, disk diffusion technique for antimicrobial resistance, serotyping, phylogeny by PCR / Analysis and publication of results
Virotype and antimicrobial resistance of E. coli isolates from pigs and chickens on farm and at slaughter in Vietnam / National Institute of Veterinary Research (NIVR), Vietnam / 2008-ongoing / Demonstration of unique virotypes and antimicrobial resistance patterns in Vietnam (see section 12 for publications) / Survey of food born potentially zoonotic E. coli in pigs and chickens
Virotype and antimicrobial resistance of E. coli isolates from pigs with diarrhoea in South Korea / National Veterinary Research and Quarantine Service(NVRQS), South Korea / 2009-ongoing / O serotyping of 150 isolates / Analysis and publication of results
Virotype and antimicrobial resistance of E. coli isolates from chickens on farm and at slaughter in Sénégal / École Inter-États des Sciences et Médecine Vétérinaires (EISMV), Sénégal / 2011-ongoing / Identification of potentially pathogenic E. coli and their antimicrobial resistance / Continued characterisation of isolates, analysis, publication of 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.
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 stayMSc 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.
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.