OIE Reference Laboratory Reports
Activities in 2011
Name of disease for which you are Salmonellosis
a designated OIE Reference
Address of laboratory: OIE Reference Laboratory for Salmonellosis
Laboratory for Foodborne Zoonoses
Public Health Agency of Canada
110 Stone Road West, Guelph, Ontario
N1G 3W4 CANADA
e-mail address: firstname.lastname@example.org
Name of Head of Laboratory: Dr Roger Johnson
Name of OIE Reference Expert: Dr Cornelis Poppe
Annual reports of OIE Reference Centres, 2011 1
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
The OIÉ Reference Laboratory for Salmonellosis is a national Salmonella surveillance centre, contributing
laboratory support, Salmonella cultures and submissions data to the national enteric pathogen surveillance system
of the Public Health Agency of Canada, the Canadian Integrated Program for Antimicrobial Resistance
(http://www.phac-aspc.gc.ca/cipars-picra/index.html), and PulseNet Canada (the Canadian listserve for
PFGE/molecular methods, bacterial outbreak investigations and surveillance). The Laboratory also supports
provincial government Salmonella monitoring programs and research programs of government institutions and
universities in Canada and internationally.
Serotyping: 6323 Salmonella cultures isolated from animals and their environment, foods, feeds and other sources
in Canada during the year 2011 were identified serologically using the antigenic formulae of Grimont and Weill
(2007) and are shown in Table 1.
Table 1. The commonest Salmonella serovars and number of isolates of each serovar
Rank Serovar Number Rank Serovar Number Rank Serovar Number
1 Kentucky 1616 5 Hadar 217 9 Mbandaka 123
2 Enteritidis 883 6 Infantis 210 10 Derby 120
3 Heidelberg 871 7 Senftenberg 126 11 Schwarzengrund 118
4 Typhimuriuma 366 8 Orion var. 124 12 Other serovars 1549
Includes S. Typhimurium (154 isolates) and S. Typhimurium var. Copenhagen (212 isolates)
Table 2. The commonest Salmonella serovars and number of isolates of each serovar from each species or source
Specie Commonest serovars and number of isolates of each serovar from each Total
Chicken Kentucky 1101 Heidelberg 496 or source
Enteritidis 496species Hadar 96 isolate
Food - Kentucky 345 Heidelberg 215 Enteritidis 178 Hadar 36 953
Porcine Typhimuriuma 120 Derby 83 Infantis 45 Worthington 27 528
Avianb Enteritidis 96 Heidelberg 64 Kentucky 35 Hadar 27 426
Turkey Orion var. 74 Schwarzengrund 45 Senftenberg 42 Hadar 36 382
Bovine Typhimuriumc 97 I:6,14,18:-:- 10 Enteritidis 9 Dublin 7 181
Includes S. Typhimurium (40 isolates) and S. Typhimurium var. Copenhagen (80 isolates); b Avian species other
than chicken and turkey; c Includes S. Typhimurium (27 isolates) and S. Typhimurium var. Copenhagen (70
Phagetyping: The Salmonella Heidelberg, S. Enteritidis and S. Typhimurium strains were phagetyped with the
typing schemes of Demczuk et al. (J Clin Microbiol 2003; 41:4279-4284), Ward et al. (Epidemiol Infect 1987;
99:291-294), and Anderson et al. (J Hyg 1977; 78:297-300), respectively. A total of 2,193 strains were
phagetyped. The results are shown in Table 3.
2 Annual reports of OIE Reference Centres, 2011
Table 3. The commonest phagetypes of S. Heidelberg, S. Enteritidis and S. Typhimurium
Salmonella serovar PT No. PT No. PT No. PT No. Total
Heidelberg 19 280 29 137 18 91 Other 349 857
Enteritidis 8 310 13a 257 13 70 Other 217 854
Typhimurium var. Copenhagen c UT1 29 104 18 UT5 16 Other 184 247
Typhimurium b 104 30 108 26 132 17 Other 162 235
PT denotes phagetype; b Includes S. Typhimurium var. Copenhagen-like strains with the antigenic formula:
1,4,12:i:-. c Includes S. Typhimurium-like strains with the antigenic formula: 1,4,5,12:i:-;
Four strains of Salmonella Enteritidis PT4 were identified.
2. Production and distribution of diagnostic reagents
Cultures are available from the Salmonella strain collection (which consists of more than 60,000 strains) for
research and diagnostic purposes at laboratories nationally and in other OIE Member countries.
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
The OIÉ Reference Laboratory has been a member of the WHO Global Foodborne Infections Network
(who.int/gfn/en) since 2000 and is listed on its web page (who.int/gfn/members/en). The laboratory tested a
blind panel of 100 Salmonella serovars from the Veterinary Laboratories Agency in the United Kindom using
a Salmonella geno-serotyping array (SGSA) (see Section 12 below).
b) Organisation of inter-laboratory proficiency testing with laboratories other than OIE
Reference Laboratories for the same pathogens and diseases to ensure equivalence of
The Laboratory has successfully participated in a yearly External Quality Assurance System (EQAS) for
Salmonella serotyping with other Global Foodborne Infections Network member laboratories since 2001.
4. Preparation and supply of international reference standards for diagnostic tests or vaccines
Salmonella antisera, Salmonella cultures and other diagnostic supplies are available to researchers and
diagnosticians in Member Countries.
5. Research and development of new procedures for diagnosis and control
The following topics are being studied: The epidemiology, pathogenesis and control of salmonellosis in animals
and humans; antimicrobial resistance and trends in resistance of Salmonella; the association between antimicrobial
resistance and virulence in Salmonella; further characterization of Salmonella by serotyping, phagetyping, PFGE,
plasmid profiles, and by PCR to identify antimicrobial resistance, virulence and other genes; sequencing of DNA
and other molecular typing and research techniques; alternative molecular methods to traditional serotyping. The
Annual reports of OIE Reference Centres, 2011 3
laboratory led an international scientific collaborative study to develop a Salmonella geno-serotyping array
(SGSA) that rapidly generates an antigenic formula consistent with the White-Kauffmann-Le Minor scheme (see
Section 12 and Reference 1). A genomic and proteomic characterization was carried out on the Salmonella
bacteriophage PVP-SE1; the phage has a broad lytic spectrum against different Salmonella serovars (see
Reference 4). Two Salmonella enterica serovar Typhimurium-based vaccines against necrotic enteritis were
assessed to determine concomitant reduction in colonization of chickens by Salmonella serovars of different
serogroups (see Reference 12).
6. Collection, analysis and dissemination of epizootiological data relevant to international disease
A recent study showed that Salmonella Enteritidis has emerged as the most prevalent cause of human
salmonellosis in Canada. Recent trends of S. Enteriditis subtypes and their potential sources were described by
integrating Salmonella data from several Canadian surveillance and monitoring programmes. A threefold increase
in S. Enteritidis cases from 2003 and 2009 was primarily associated with phage types 13, 8 and 13a. Other
common phage types (4, 1 and 6a) showed winter seasonality and were more likely associated with cases linked to
international travel. Conversely, phage types 13, 8 and 13a had summer seasonal peaks and were associated with
cases of domestically acquired infections. During agri-food surveillance, S. Enteriditis was detected in various
commodities, most frequently in chicken (with phage types 13, 8 and 13a predominating). Antimicrobial
resistance was low in human and non-human isolates. Continued integrated surveillance and collaborative
prevention and control efforts are necessary to mitigate future illness (see Reference 17).
7. Maintenance of a system of quality assurance, biosafety and biosecurity relevant to the
pathogen and the disease concerned
The OIÉ Reference Laboratory for Salmonellosis has been accredited by the Standards Council of Canada since
1997 and its diagnostic procedures comply with the requirements of the ISO/IEC Guide 17025. Besides
participating in the yearly External Quality Assurance System (EQAS) for Salmonella serotyping with other
Global Foodborne Infections Network member laboratories (see Section 3b), the laboratory continued its
participation in a joint inter-laboratory quality assurance (QA) program with the Ontario Agency for Health
Protection and Promotion, Toronto, Ontario, Canada and the Laboratoire d’épidémiosurveillance animale du
Québec, Saint-Hyacinthe, Québec, Canada. It also carried out an internal QA program. A total of 270 isolates
belonging to 78 serovars unique to the QA program and to other more commonly occurring serovars were
serotyped, and 16 strains were phagetyped as part of the QA program.
8. Provision of consultant expertise to OIE or to OIE Member Countries
The OIE Expert on Salmonellosis at our Laboratory completed an OIE questionnaire on salmonellosis in
connection with an attempt to prioritize animal diseases in East Africa. He answered inquiries from OIE member
countries about procedures and protocols regarding the isolation, identification, virulence, antimicrobial resistance
and host-specificity of Salmonella isolates.
9. Provision of scientific and technical training to personnel from other OIE Member Countries
10. Provision of diagnostic testing facilities to other OIE Member Countries
The OIE Salmonellosis Reference Laboratory has had a long-standing relationship with researchers and
epidemiologists in Ethiopia and serotyped and determined antimicrobial resistance of bovine, human and
environmental Salmonella strains isolated in Ethiopia (see Reference 22).
11. Organisation of international scientific meetings on behalf of OIE or other international bodies
4 Annual reports of OIE Reference Centres, 2011
12. Participation in international scientific collaborative studies
The laboratory led an international scientific collaborative study to develop a Salmonella geno-serotyping array
(SGSA) that rapidly generates an antigenic formula consistent with the White-Kauffmann-Le Minor scheme,
currently the gold standard for Salmonella serotyping. A set of 287 strains representative of 133 Salmonella
serovars was assembled to validate the array and to test the array probes for accuracy, specificity, and
reproducibility. Initially, 76 known serovars were utilized to validate the specificity and repeatability of the array
probes and their expected probe patterns. The SGSA generated the correct serovar designations for 100% of the
known subspecies I serovars tested in the validation panel and an antigenic formula consistent with that of the
White-Kauffmann-Le Minor scheme for 97% of all known serovars tested. Once validated, the SGSA was
assessed against a blind panel of 100 Salmonella enterica subsp. I samples serotyped using traditional methods
correctly identifying all samples as Salmonella, and successfully identifying 92% of the antigens found within the
unknown samples. Antigen- and serovar-specific probes, in combination with a pepT PCR for confirmation of S.
enterica subsp. enterica determinations, generated an antigenic formula and/or a serovar designation consistent
with the White-Kauffmann-Le Minor scheme for 87% of unknown samples tested with the SGSA. Although the
current array has been validated for detection of the 76 target serovars it is important to note that the combination
of probes on the current layout has the potential to type 1026 subspecies I. A tri-site collaborative validation of
the SGSA is currently underway involving testing of >770 strains representing >150 serovars to determine the
specificity, sensitivity, repeatability and robustness of the assay. This will be followed by parallel testing of
approximately 700 incoming Salmonella isolates at each of the three locations in an effort to demonstrate the
versatility and utility of this array as a public health tool in the identification of Salmonella (see Reference 1). To
increase the detection capabilities of future array layouts the group has sequenced more than 20
Salmonella isolates which are undergoing assembly, gap closure and annotation. An additional 40 strains are
scheduled for sequencing in 2012 which will include representatives from all previously unsequenced serogroups,
and allow for differentiation of highly homologous serotypes.
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
1. Yoshida C, Franklin K, Lingohr E, Kostic T, Anjum M, Clark C, Kropinski A. Blind assessment of human,
animal and environmental samples using the Salmonella Geno-Serotyping Array (SGSA). FEMS, Geneva,
Switzerland. June 26-30, 2011. Poster Presentation.
2. Yoshida C, Lingohr EJ, Franklin K, Bodrossy L, Anjum M, Clark CG, Kropinski AM. A Salmonella Geno-
Serotyping Array (SGGSA) for the rapid classification of serovars. National Molecular Microbiology
Diagnostic Users Group Annual Fall Meeting (NMG), Toronto, Canada. October 17-18, 2011. Oral
Scientific publications in peer-reviewed journals
1. Franklin K, Lingohr EJ, Yoshida C, Anjum M, Bodrossy L, Clark CG, Kropinski AM, Karmali MA. Rapid
genoserotyping tool for classification of Salmonella serovars. J Clin Microbiol 2011; 49:2954-2965.
2. Laing C, Villegas A, Taboada EN, Kropinski, A, Thomas JE, Gannon VP. Identification of Salmonella
enterica species- and subgroup-specific genomic regions using Panseq 2.0. Infect Genet Evol 2011; 11:2151-
3. Anany H, Lingohr EJ, Villegas A, Ackermann H-W, She Y-M, Griffiths MW, Kropinski AM. A Shigella
boydii bacteriophage which resembles Salmonella phage ViI. Virology Journal 2011; 8:242.
4. Santos SB, Kropinski AM, Ceyssens PJ, Ackermann H-W, Villegas A, Lavigne R, Krylov VN, Carvalho CM,
Ferreira EC, Azeredo J. Genomic and proteomic characterization of the broad-host-range Salmonella phage
PVP-SE1: creation of a new phage genus. J Virol 2011; 85:11265-11273.
5. Kropinski AM, Arutyunov D, Foss M, Cunningham A, Ding W, Singh A, Pavlov AR, Henry M, Evoy S,
Kelly J, Szymanski CM. Genome and proteome of Campylobacter jejuni bacteriophage NCTC 12673. Appl
Environ Microbiol 2011; 77:8265-8271.
Annual reports of OIE Reference Centres, 2011 5
6. Kutter EM, Skutt-Kakaria K, Blasdel B, El-Shibiny A, Castano A, Bryan D, Kropinski AM, Villegas A,
Ackermann H-W, Toribio AL, Pickard D, Anany H, Callaway T, Brabban AD. Characterization of a ViI
phage specific to Escherichia coli O157:H7. Virology Journal 8:430.
7. Martin LC, Weir EK, Poppe C, Reid-Smith RJ, Boerlin P. Characterization of blaCMY-2 plasmids in
Salmonella and E. coli from food animals in Canada. Appl Environ Microbiol 2011; Dec 9 [Epub ahead of
8. Jardine C, Reid-Smith RJ, Janecko N, Allan M, McEwen SA. Salmonella in raccoons (Procyon lotor) in
southern Ontario, Canada. J Wildl Dis 2011; 47:344-351.
9. Allen SE, Boerlin P, Janecko N, Lumsden JS, Barker IK, Pearl DL, Reid-Smith RJ, Jardine C. Antimicrobial
resistance in generic Escherichia coli isolates from wild small mammals living in swine farm, residential,
landfill, and natural environments in southern Ontario, Canada. Appl Environ Microbiol 2011; 77:882-888.
10. Cook A, Reid-Smith RJ, Irwin RJ, McEwen SA, Young V, Ribble C. Antimicrobial resistance in
Campylobacter, Salmonella, and Escherichia coli isolated from retail grain-fed veal meat from Southern
Ontario, Canada. J Food Prot 2011; 74:1245-1251.
11. Mather AE, Matthews L, Mellor DJ, Reeve R, Denwood MJ, Boerlin P, Reid-Smith RJ, Brown DJ, Coia JE,
Browning LM, Haydon DT, Reid SW. An ecological approach to assessing the epidemiology of antimicrobial
resistance in animal and human populations. Proc Biol Sci 2011; Nov 16 [Epub ahead of print].
12. Jiang Y, Kulkarni RR, Parreira VR, Poppe C, Roland KL, Prescott JF. Assessment of 2 Salmonella enterica
serovar Typhimurium-based vaccines against necrotic enteritis in reducing colonization of chickens by
Salmonella serovars of different serogroups. Can J Vet Res 2010; 74:264-270.
13. Farzan A, Friendship RM, Dewey CE, Poppe C, Funk J. Evaluation of the risk factors for shedding
Salmonella with or without antimicrobial resistance in swine using multinomial regression method. Zoonoses
Public Health 2010; 57: Suppl 1:85-93.
14. Garcia-Migura L, Sunde M, Karismose S, Veldman K, Schroeter A, Guerra B, Granier SA, Perrin-Guyomard
A, Gicquel-Bruneau M, Franco A, Englund S, Teale C, Heiska H, Clemente L, Boerlin P, Moreno MA,
Daignault D, Mevius D, Hendriksen RS, Aarestrup FM. Establishing streptomycin epidemiological cut-off
values for Salmonella and Escherichia coli. Microb Drug Resist 2011; Jul 12 [Epub ahead of print].
15. Jokinen C, Edge TA, Ho S, Koning W,Laing C, Mauro W, Medeiros D, Miller J, Robertson W, Taboada E,
Thomas JE, Topp E, Ziebell K, Gannon VP. Molecular subtypes of Campylobacter spp., Salmonella enterica,
and Escherichia coli O157:H7 isolated from faecal and surface water samples in the Oldman river watershed,
Alberta, Canada. Water Res 2011; 45:1247-1257.
16. Wilkes G, Edge TA, Gannon VP, Jokinen C, Lyautey E, Neumann NF, Ruecker N, Scott A, Sunohara M,
Topp E, Lapen DR. Associations among pathogenic bacteria, parasites, and environmental and land use
factors in multiple mixed-use watersheds. Water Res 2011; 45:5807-5825.
17. Nesbitt A, Ravel A, Murray R, McCormick R, Savelli C, Finley R, Parmley J, Agunos A, Majowicz SE,
Gilmour M and the Canadian Integrated Program for Antimicrobial Resistance Surveillance Public Health
Partnership and the Canadian Public Health Laboratory Network. Integrated surveillance and potential sources
of Salmonella Enteritidis in human cases in Canada from 2003 to 2009. Epidemiol Infect 2011; Dec 14:1-16
[Epub ahead of print].
18. Davidson VJ, Ravel A, Nguyen TN, Fazil A, Ruzante JM. Food-specific attribution of selected
gastrointestinal illnesses: estimates from a Canadian expert elicitation survey. Foodborne Pathog Dis 2011;
19. Ruzante JM, Majowicz SE, Fazil A, Davidson VJ. Hospitalization and deaths for select enteric illnesses and
associated sequelae in Canada, 2001-2004. Epidemiol Infect 2011; 139:937-945.
20. Bucher O, Fazil A, Rajić A, Farrar A, Wills R, McEwen SA. Evaluating interventions against Salmonella in
broiler chickens: applying synthesis research in support of quantitative exposure assessment. Epidemiol Infect
2011; Jul 25:1-21 [Epub ahead of print].
6 Annual reports of OIE Reference Centres, 2011
21. Bucher O, Farrar AM, Totton SC, Wilkins W, Waddell LA, Wilhelm BJ, McEwen SA, Fazil A, Rajić A. A
systematic review-meta-analysis of chilling interventions and a meta-regression of various processing
interventions for Salmonella contamination of chicken. Prev Vet Med 2012; 103:1-15 [Epub 2011 Oct 10].
22. Sibhat B, Molla Zewde B, Zerihun A, Muckle A, Cole L, Boerlin P, Wilkie E, Perets A, Mistry K, Gebreyes
WA. Salmonella serovars and antimicrobial resistance profiles in beef cattle, slaughterhouse personnel and
slaughterhouse environment in Ethiopia. Zoonoses Public Health 2011; 58:102-109.
23. Leonard EK, Pearl DL, Finley RL, Janecko N, Reid-Smith RJ, Peregrine AS, Weese JS. Comparison of
antimicrobial resistance patterns of Salmonella spp. and Escherichia coli recovered from pet dogs from
volunteer households in Ontario (2005-06). J Antimicrob Chemother 2012; 67:174-181 [Epub 2011 Oct 20].
Annual reports of OIE Reference Centres, 2011 7