OIE Reference Laboratory Reports
Activities in 2011
Name of disease (or topic) for
which you are a designated OIE
Reference Laboratory:
Bovine spongiform encephalopathy (BSE)
Address of laboratory:
National Centre for Animal Diseases /Canadian Food Inspection
Agency
Lethbridge Laboratory
P.O.Box 640 (Township Road 9-1)
Lethbridge/Alberta T1J 3Z4
Canada
Tel.:
+1(403) 382-5549
Fax:
+1(403) 382-5583
e-mail address:
website:
Name (including Title and
Position) of Head of Laboratory
(Responsible Official):
Name(including Title and
Position) of OIE Reference
Expert:
stefanie.czub@inspection.gc.ca
www.cfia.ca
Dr Soren Alexandersen, Executive Director
National Centres for Animal Diseases
(NCAD)/CFIA
NCFAD-Winnipeg & ADRI-Lethbridge
Dr. Stefanie Czub, Research Manager;
Head of the National & OIE Reference
Laboratories for BSE
Name (including Title and
Position) of writer of this report
(if different from above):
Annual reports of OIE Reference Centres, 2011
1
Bovine spongiform encephalopathy
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 Canadian reference laboratory (RL) performs Immunohistochemistry (IHC) and/or the SAF Immunoblot for
the confirmation of BSE. In addition, histology - pertaining on the tissue quality -, the Hybrid Western Blot and
several rapid tests for BSE are part of the confirmatory schedule. Different rapid tests (n=6) are implemented and
one is used for BSE surveillance within the laboratory. The laboratory is also part of the national Scrapie
surveillance network.
Test
For
Specificity
Total (number of cases)
5
Antigen, Surveillance
BSE
9627
IHC
Antigen, Confirmation
BSE
5
Histology
Morphological changes,
confirmation
BSE
5
SAF Immunoblot
Antigen, Confirmation
BSE
1
Hybrid Western Blot
Antigen, Confirmation
BSE
1
BioRad TeSeE Elisa
Antigen, Confirmation
BSE
1
Prionics-Check Western
Antigen, Confirmation
BSE
1
Prionics-Check PrioStrip
Antigen, Confirmation
BSE
1
The Canadian RL performs the batch release testing of individual rapid tests series used by the Canadian
network laboratories for BSE surveillance, including one ELISA and one lateral flow immunoassay. Panels
consist of 16 or 18 members respectively; the BSE positive and negative panel members are from cattle of
Canadian origin.
2.
Production and distribution of diagnostic reagents
For quality assurance and quality control purposes, proficiency panels for BSE rapid tests (biannually) and IHC
(annually) are distributed nationally to the laboratories of the Canadian BSE network (n=8) and internationally to
BSE reference laboratories (n=5). The panel consists of Scrapie-positive and bovine negative brain homogenates
(n=6) for the respective rapid tests and of unstained Chronic Wasting Disease- and Scrapie-positive and unstained
bovine negative brain sections for the IHC.
Type of reagent
Amount supplied nationally
(including for own use)
Amount supplied
internationally
Tissue homogenates
93 ml
33 ml
Tissue sections
140
30
Tissue blocks
4
12
Blood samples
N/A
N/A
Urine samples
N/A
N/A
Fresh tissue
5g
60 g
Tissues, blood and other samples from experimentally (i.c.) infected C-, L- and H-type cattle are available for
interested parties upon request.
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Annual reports of OIE Reference Centres, 2011
Bovine spongiform encephalopathy
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
 International co-operations exist with the Argentinean, British, Japanese and Swiss OIE BSE Reference
Laboratories on exchanging reference materials, harmonising IHC methods and pursuing research
projects on atypical BSE.
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
 As part of the ISO 17025 certification, the Canadian RL participates in ring trials for BSE surveillance
and BSE IHC provided by the German TSE reference Laboratory (Friedrich-Loeffler-Institute). The ring
trials monitor the laboratory’s performance in a rapid western blot, a lateral flow immunoassay and in one
ELISA. In addition, proficiency panels for the performance of the IHC were received.
 For quality assurance and quality control purposes, proficiency panels for BSE rapid tests and IHC are
distributed biannually or annually to the national reference laboratories of Austria, Australia, China,
Germany and the USA. The panel consists of Scrapie-positive and bovine negative brain homogenates
(n=6) for the respective rapid tests and of unstained Chronic Wasting Disease and Scrapie-positive and
unstained bovine negative brain sections for the IHC.
4.
5.
Preparation and supply of international reference standards for diagnostic tests or vaccines
Type of reagent
Amount available
Amount supplied
nationally
(including for own use)
Amount supplied to
other countries
Tissue homogenates
N/A
93 ml
33 ml
Tissue sections
N/A
140
30
Tissue blocks
242
4
12
Blood samples
100 l
N/A
N/A
Urine samples
20 l
N/A
N/A
Fresh tissue
1700 g
N/A
60 g
Research and development of new procedures for diagnosis and control
For the purpose of risk assessment and development of control measures for atypical BSE, six and thirteen
months-old cattle respectively were intracranially and perorally challenged with atypical BSE. Samples are
collected from these animals through-out the incubation period of the disease. For the ic challenge, animals were
allowed to progress to clinical disease after which an post mortem examination was performed to collect numerous
samples. Animals are monitored monthly using a clinical examination template for the evaluation of neurological
disease in atypical BSE. This template was developed in collaboration with University of Calgary’s Veterinary
School and is to be adapted for field use for consistent recording of clinical observations. Cattle intracranial
Annual reports of OIE Reference Centres, 2011
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Bovine spongiform encephalopathy
challenged with atypical BSE had a shorter clinical period and disease progression was faster as compared to
classical BSE. The experiment is ongoing.
For diagnostic measures, a project for the detection of biomarker in bovine urine is ongoing. A small number of
proteins in the CSF and blood have been proposed as biomarkers for prion infection. Unfortunately, insufficient
sensitivity and specificity of tests based on these proteins and the invasive procedure required to obtain CSF has
limited their use in universal surveillance. We have compared and contrasted the urine protein profiles of BSEinfected and control animals over the course of the disease. Proteins demonstrating differing concentrations
between the two sample pools have been identified. Further analysis is underway to determine how sensitive and
specific these proteins are for BSE diagnosis. Two urine proteins have been demonstrated to differentiate TSE
infected mice and bovine, immunohistochemistry validation of western blot data is underway.
6.
Collection, analysis and dissemination of epizootiological data relevant to international disease
control
For the purpose of epidemiological surveillance, from January 1 st to December 31st data were collected, analyzed
and provided via BSE surveillance testing (n = 33,457). The RL confirmed one case of BSE in 2011. For the
complete testing numbers of the Canadian BSE network please see the CFIA homepage.
7.
Maintenance of a system of quality assurance, biosafety and biosecurity relevant to the
pathogen and the disease concerned
Since its designation in 2005, the BSE OIE RL is ISO 17025 accredited and operates under a Quality Management
System covering all laboratory activities that fall under the scope of accreditation, identifying policies, protocols,
and procedures designed to conform to the requirements of ISO/IEC 17025: 2005 (E), "General requirements for
the competence of calibration and testing laboratories". To ensure quality, diagnostic tests for BSE are
standardized (via test validation; use of standard reference materials; standardization of reagents and instruments/
equipment; initial certification and annual re-certification of analysts); control samples and standard protocols are
used. Key activities include documentation and document control; QC procedures for tests and equipment; internal
and external audits; and proficiency testing.
Surveillance, confirmatory and research activities in the BSE Reference Laboratory are all carried out under
Biosafety Level 3 conditions. The laboratory is regularly recertified.
8.
Provision of consultant expertise to OIE or to OIE Member Countries
Expertise to the OIE was provided via the participation in the second Global Conference of OIE Reference
Laboratories and Collaborating Centers in Paris, France; the review of the OIE BSE chapter; email contacts; and
scientific and technical training on-site and in-house to OIE member countries including Argentina, Brazil, China,
Cuba, Costa Rica (see also: 9)
Advice was provided to a Chinese delegation from the Administration of Quality Supervision, Inspection and
Quarantine (AQSIQ) department of the government of China, as part of an audit on deboned UTM beef; to staff
members of the University of Beijing’s Faculty of Agriculture on the laboratory’s role in animal disease
surveillance using BSE as a model.
On-site consultation and advice was provided to the Beijing AQSIQ on quality assurance of test kits; and potential
research collaborations with the Chinese TSE reference laboratory (Agricultural University/Beijing) were
identified during a visit to China.
9.
Provision of scientific and technical training to personnel from other OIE Member Countries
One staff member from the Argentinean BSE Reference laboratory spend several days in the laboratory and was
trained in BSE confirmatory tests including different Western Blots and Immunohistochemistry (May).
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Annual reports of OIE Reference Centres, 2011
Bovine spongiform encephalopathy
The head of the BSE Reference Laboratory and two staff member went to the Chinese Jilin and Chongqing
provinces to provide training focused on diagnostic, quality assurance and occupational issues. The trip was
planned and organized with the help of Chinese provincial, municipal and federal authorities (June).
A group of four Indonesian scientists were trained in BSE surveillance and confirmation testing; Quality
Assurance and Biocontainment Level 3 laboratory operational requirements and working practices (July).
10. Provision of diagnostic testing facilities to other OIE Member Countries
Confirmatory testing on several bovine samples was provided to SENASA/Costa Rica.
11. Organisation of international scientific meetings on behalf of OIE or other international bodies
N/A
12. Participation in international scientific collaborative studies
International scientific collaboration with the German TSE Reference Laboratory (Friedrich-Loeffler-Institute) on
the identification of Specific Risk Material and age-related susceptibility in cattle per orally challenged with
German H- and L-type BSE (Ongoing project).
International scientific collaboration with the Swiss & OIE BSE/TSE Reference Laboratory (Neurocenter Berne)
on the infectious potential of two atypical Swiss cases. Cattle were challenged intracranially with Swiss
homogenates and are presently under observation in the Lethbridge laboratory (New project).
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
S Dudas, T Pickles, Y Fang, C Graham, S Czub, M Czub: Exploring the role of dendritic cells in BSE oral uptake
and neuro-invasion. Prion2011, Montreal/Canada, May 16-19, 2011.
J Yang, S Dudas, R Flitton, T Pickles, K Colwell, Y Fang, C Graham, S Czub: Early Detection of PrP CWD in Rapid
Test-Negative Elk Using a Modified Western Blot. Prion2011, Montreal/Canada, May 16-19, 2011.
S Braithwaite, BH Gilroyed, T Reuter, S Czub, C Graham, A Balachandran, T McAllister, M Belosevic, N
Neumann: Detection of prions in anaerobic digestion sludge by PMCA. Prion2011, Montreal/Canada, May 16-19,
2011.
D Motzkus, W Schulz-Schaeffer, M Beekes, H Schaetzl, FR Jirik, AC Schmaedicke, M Daus, J Breyer, SV Gilch,
E Comoy, C Graham, JP Deslys, JA Richt, B Race, S Czub: Transmission of of CWD to Non-Human Primates:
Interim Results of a Comprehensive Study on the Transmissibility to Humans. Prion2011, Montreal/Canada, May
16-19, 2011.
L Lamoureux, SL Simon, MM Plews, C Graham, SV Ruddat, S Czub, D Knox: Factors affecting the accuracy of
urine based biomarkers of BSE. Prion2011, Montreal/Canada, May 16-19, 2011.
BH Gilroyed, T Reuter, S Braithwaite, S Czub, C Graham, A Balachandran, T McAllister, M Belosevic, N
Neumann: Prion Detection during Anaerobic Digestion of Specified Risk Materials using Protein Misfolding
Amplification (PMCA) and Immuno-Quantitative Real-Time PCR (iQ-RT-PCR). Prion2011, Montreal/Canada,
May 16-19, 2011.
C Graham, M Levy, E Pajor, G McGregor, R Flitton, S Czub: Clinical disease in cattle experimentally inoculated
with all types of BSE. Prion2011, Montreal/Canada, May 16-19, 2011.
Annual reports of OIE Reference Centres, 2011
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Bovine spongiform encephalopathy
C Graham, B Hills, M Groschup, S Dudas, S Czub: Pathogenesis and agent distribution of here known types of
BSE. Prion2011, Montreal/Canada, May 16-19, 2011.
SL Simon, L Amoureux, I Tayyab, V Ruddat, C Graham, S Czub, D Knox: The specificity of Scrapie-induced
changes in the murine urine proteome. Prion2011, Montreal/Canada, May 16-19, 2011.
 Scientific publications in peer-reviewed journals
Simmons MM, Spiropoulos J, Webb P, Spencer YI, Czub S, Mueller R, Davis A, Arnold ME, Marsh S, SAC
Hawkins, Cooper JA, Konold T, Wells GAH. Experimental classical bovine spongiform encephalopathy:
definition and progression of neural PrP immunolabeling in relation to diagnosis and disease controls. Vet Pathol,
48: 948-963, 2011.
M Plews, L Lamoureux, SLR Simon, C Graham, V Ruddat, S Czub, J D Knox: Factors affecting the accuracy
of urine-base biomarkers of BSE. Proteome Science, 9-6, 2011.
JG Gray, S Dudas, S Czub: A Study on the Analytical Sensitivity of 6 BSE Tests Used by the Canadian BSE
Reference Laboratory. PLOS One, 2011, 6,3, 1-10;e17633.

Other communications
N/A
_______________
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Annual reports of OIE Reference Centres, 2011