Brucellosis
Klaus Nielsen
Ottawa Laboratories (Fallowfield), Canadian Food Inspection Agency
3851 Fallowfield Road, Ottawa, Ontario, Canada K2H 8P9
Tel.: (1-613) 228-6698, Fax: (1-613) 228-6669
nielsenk@inspection.gc.ca
Summary of general activities related to the disease
1.
Test(s) in use/or available for the specified disease at your laboratory
Test
For
Specificity
Total
BPAT
Antibody
Whole cell
20093
CFT
Antibody
Whole cell
7901
ELISA
Antibody
LPS
5220
FPA
Antibody
OPS
2856
TAT
Antibody
Whole cell
6083
CFT B. ovis
Antibody
Whole cell
645
Bacterial culture
Isolation
NA
87 animals
PCR
Identification
NA
171 items
Bacterial cultures were attempted on 120 tissues from 73 marine mammals and tissues from 12 caribou in support
of research for a total of 324 set-ups. Samples from 2 caribou were cultured for diagnostic purposes. Eleven
Brucella sp. were biotyped.
171 tissues/bacterial colonies were tested by various PCR procedures
2.
Production and distribution of diagnostic reagents
BPAT antigen (63700 doses) was produced for use at CFIA and contract laboratories.
TAT and CFT (B. abortus and B. ovis) whole cell antigens (30000 doses total) were produced for use at the CFIA
Ottawa and Lethbridge laboratories and for human diagnostics.
SLPS was prepared for CELISA and OPS-FITC for FPA use at the Ottawa laboratory.
Diagnostic reagents were distributed to the Departments of Agriculture in Argentina, Chile and China.
Reagents for research purposes were distributed to the Federal and/or State Departments of Agriculture/
Universities of Argentina, Brazil, Chile, China, Kazakhstan, Mexico and USA.
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
Provision of standardized reagents for ELISAs and FPA. Internal QC reagents for certification. Participation in
testing proposed small ruminant and porcine brucellosis standard reference sera.
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2007
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Brucellosis
4.
Preparation and supply of international reference standards for diagnostic tests or vaccines
Prepared a candidate serum to be considered as the standard reference serum for porcine brucellosis. The serum
was distributed to other OIE reference labs for testing. Results are pending (analysis by the OIE Collaborating
Center at Weybridge)
5.
Research and development of new procedures for diagnosis and control
Monoclonal antibodies and SLPS preparations were provided to a number of national, state and university
laboratories for the IELISA and the CELISA.
O-polysaccharide and R-core antigens labelled with fluorescein isothiocyanate was provided for FPA use to a
number of laboratories.
6.
Collection, analysis and dissemination of epizootiological data relevant to international disease
control
Collaborating with the M. Aikimbayev’s Kazakh Scientific Center for Quarantine and Zoonotic Diseases via a
project (Dr. T. Grushina, leader) to establish prevalence of brucellosis in areas of southern Kazakhstan and to
transfer current diagnostic methods.
A large number of marine mammals were tested for the presence of antibodies in blood and cavity fluids in
collaboration with the Department of Fisheries and Ocean, British Colombia provincial government, University of
Guelph, the Nunavik Research Center and other establishments. A number of tissues were cultured for isolation of
Brucella sp. PCR was performed on numerous tissues for detection of Brucella sp.
PCR assays for detection of Brucella sp. and each individual species as well as S19 and RB51 were adapted for
maximum sensitivity and fine tuned for distinguishing terrestrial and marine isolates of Brucellae.
Collaborated with Department of Fisheries and Oceans, Government of Canada and the Government of the
Northwest Territories on the dangers of marine mammal brucellosis to Native Canadians.
7.
Provision of consultant expertise to OIE or to OIE Member Countries
Consulted with the China Animal Health and Epidemiology Center, Qindao. Participated in and acted as reporter
for ad hoc meeting on brucellosis at the OIE headquarters.
Provided data for inclusion of the FPA for the serological diagnosis of brucellosis in cattle as a prescribed test for
international trade and for the FPA for the serological diagnosis of brucellosis in small ruminants as an alternative
test for international trade.
Consulted with the M. Aikimbayev’s Kazakh Scientific Center for Quarantine and Zoonotic Diseases, Almaty,
Kazakhstan.
8.
Provision of scientific and technical training to personnel from other OIE Member Countries
Drs. Xu Wen Jiang and Jaibo Ding, China Institute of Veterinary Drug Control visited the laboratory to receive an
update on diagnostic methodology, both serological and molecular biology procedures.
9.
Provision of diagnostic testing facilities to other OIE Member Countries
NA
10. Organisation of international scientific meetings on behalf of OIE or other international bodies
NA
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Annual reports of OIE Reference Laboratories and Collaborating Centres, 2007
Brucellosis
11. Participation in international scientific collaborative studies
Collaborating with the M. Aikimbayev’s Kazakh Scientific Center for Quarantine and Zoonotic Diseases via a
project (Dr. T. Grushina, leader) to establish prevalence of brucellosis in areas of southern Kazakhstan and to
transfer current diagnostic methods.
Collaborated with the China Animal Health and Epidemiology Center, Qindao (Dr. W. Fan and colleagues).
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
Yu WL, Nielsen K, Grushina T, Meka-Mechanko. Diagnosis of human brucellosis using AMOS PCR. Eur Soc
Clin Microbiol Infec Dis. Nice. April 2006.
Nielsen K, Smith P, Yu WL, Kelly W, Gall D, Halbert G. Rapid, accurate serological diagnosis of brucellosis.
CAHLN, Ottawa, June 2006.
Yu WL, Nielsen K. Diagnosis of brucellosis using modified AMOS PCR. CAHLN, Ottawa, June 2006.
Nielsen K, Yu WL. Quality control for serological diagnosis by enzyme immunoassay. China Epizootiology
Center, Qingdao, PRC, July 2006.
Nielsen K, Yu WL. ISO 17025 accreditiation. Overview of requirements. China Epizootiology Center, Qingdao,
PRC, July 2006.
Nielsen K, Yu WL. Control of brucellosis. China Epizootiology Center, Qingdao, PRC, July 2007.
Nielsen K, Yu WL. Diagnosis of brucellosis. China Epizootiology Center, Qingdao, PRC, July 2007.
Nielsen K, Yu WL. Vaccination against brucellosis. China Epizootiology Center, Qingdao, PRC, July 2007.
Nielsen K. Historical and contemporary aspects of diagnosis and control of brucellosis. Catedra Nacional de
Mexicana Veterinaria, Universidad Autonoma de baja California. Mexicali, Mexico October 2007.
 Scientific publications in peer-reviewed journals
Nielsen K, Smith P, Yu W, Nicoletti P, Jungersen G, Stack J, Godfroid J. Serological discrimination by indirect
enzyme immunoassay between the antibody response to Brucella sp. and Yersinia enterocolitica O:9 in cattle and
pigs. Vet. Immunol. Immunopathol. 109: 69-78, 2006.
Ramirez-Pfeiffer C, Nielsen K, Marin-Ricalde F, Rodrigues-Padilla C, Gomez-Flores R. Comparison of
fluorescence polarization assay with card and complement fixation tests for the diagnosis of goat brucellosis in a
high prevalence area. Vet. Immunol. Immunopathol. 110: 121-127, 2006.
Nielsen K, Smith P, Yu W, Nicoletti P, Elzer P, Robles C, Bermudez R, Renteria T, Moreno F, Ruiz A, Massangill
C, Muenks Q, Jurgersen G, Tollersrud T, Samartino L, Conde S, Forbes L, Gall D, Perex B, Rojas X, Minas A.
Towards single screening tests for brucellosis. Rev sci tech Off int Epiz. 24: 1027-1038, 2005
Gall D, Nielsen K, Yu W, Smith P. Rapid field-adapted indirect enzyme-linked immunosorbent assay for detection
of antibodies in bovine whole blood and serum to Brucella abortus. Clin Vaccine Immuno. l13: 501-506, 2006.
Ramirez-Pfeiffer C, Nielsen K, Marin-Ricalde F, Rodriguez-Padilla C, Gomez-Flores R. Comparison of
fluorescence polarization assay with card and complement fixation tests for the diagnosis of goat brucellosis in a
high-prevalence area. Vet Immunol Immunpathol., 110: 121-127, 2006.
Nielsen K, Smith P, Yu WL. Detection of anti-Brucella antibodies in Llama (Lama glama). J. Immunoassay
Immunochem., 28: 61-66, 2007.
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Brucellosis
Ramirez-Pfeiffer C, Nielsen K, Smith P, Marin-Ricalde F, Rodriquez-Padilla C, Gomez-Flores R. Application of
the fluorescence polarization assay for detection of caprine antibodies to Brucella melitensis in areas of high
prevalence and widespread vaccination. Clin Vaccine Immunol., 14: 299-303, 2007.
Nielsen K, Smith P, Yu WL, Halbert G. Salmonella enterica serotype Urbana interferes with brucellosis serology.
J Immunoassay Immunochem., 28:289-296, 2007.
Nielsen K, Smith P, Yu WL, Rojas X, Perez B, Conde S, Samartino L, Robles C. Detection of ovine antibody to
Brucella ovis by indirect enzyme immunoassay. J. Immunoassay Immunochem., 28: 243-250, 2007.
Nielsen K, Smith P, Yu WL, Elmgren C, Nicoletti P, Perez B, Bermudez R, Renteria T. Second generation
competitive enzyme immunoassay for detection of bovine antibody to Brucella abortus. Vet. Microbiol., 124: 173177, 2007.
Muma JB, Toft N, Oloya J, Lund A, Nielsen K, Samui K, Skjerve E. Evaluation of three serological tests for
brucellosis in naturally infected cattle using latent class analysis. Vet. Microbiol., 125: 187-192, 2007.
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Annual reports of OIE Reference Laboratories and Collaborating Centres, 2007