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Clinical and Vaccine Immunology, June 2009, p. 906-915, Vol. 16, No. 6
1071-412X/09/$08.00+0     doi:10.1128/CVI.00413-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Serological Diagnosis of Brucella Infections in Odontocetes{triangledown}

Gabriela Hernández-Mora,1 Charles A. Manire,2 Rocío González-Barrientos,1 Elías Barquero-Calvo,1 Caterina Guzmán-Verri,1 Lydia Staggs,3 Rachel Thompson,4 Esteban Chaves-Olarte,1,5 and Edgardo Moreno1*

Programa de Investigación en Enfermedades Tropicales, Escuela de Medicina Veterinaria, Universidad Nacional, Heredia, Costa Rica,1 Mote Marine Laboratory, 1600 Ken Thompson Parkway, Sarasota, Florida,2 Gulf World Marine Park, 15412 Front Beach Road, Panama City Beach, Florida 32413,3 Six Flags Discovery Kingdom, 1001 Fairgrounds Dr., Vallejo, California 94589,4 Centro de Investigación en Enfermedades Tropicales, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica5

Received 17 October 2008/ Returned for modification 12 January 2009/ Accepted 15 April 2009

Brucella ceti causes disease in Odontoceti. The absence of control serum collections and the diversity of cetaceans have hampered the standardization of serological tests for the diagnosis of cetacean brucellosis. Without a "gold" standard for sensitivity and specificity determination, an alternative approach was followed. We designed an indirect enzyme-linked immunosorbent assay (iELISA) that recognizes immunoglobulins G (IgGs) from 17 odontocete species as a single group. For the standardization, we used Brucella melitensis and Brucella abortus lipopolysaccharides, serum samples from seven resident odontocetes with no history of infectious disease displaying negative rose bengal test (RBT) reactions, and serum samples from seven dolphins infected with B. ceti. We compared the performance of the iELISA with those of the protein G ELISA (gELISA), the competitive ELISA (cELISA), and the immunofluorescence (IF) and dot blot (DB) tests, using 179 odontocete serum samples and RBT as the reference. The diagnostic potential based on sensitivity and specificity of the iELISA was superior to that of gELISA and cELISA. The correlation and agreement between the iELISA and the gELISA were relatively good (Ri/g2 = 0.65 and {kappa}i/g = 0.66, respectively), while the correlation and agreement of these two ELISAs with cELISA were low (Ri/c2 = 0.46, Rg/c2 = 0.37 and {kappa}i/c = 0.62, {kappa}g/c = 0.42). In spite of using the same anti-odontocete IgG antibody, the iELISA was more specific than were the IF and DB tests. An association between high antibody titers and the presence of neurological symptoms in dolphins was observed. The prediction is that iELISA based on broadly cross-reacting anti-dolphin IgG antibody would be a reliable test for the diagnosis of brucellosis in odontocetes, including families not covered in this study.

* Corresponding author. Mailing address: Programa de Investigación en Enfermedades Tropicales, Escuela de Medicina Veterinaria, Universidad Nacional, Heredia, Costa Rica. Phone: (506) 22380761. Fax: (506) 22381298. E-mail: emoreno{at}racsa.co.cr

{triangledown} Published ahead of print on 22 April 2009.

Clinical and Vaccine Immunology, June 2009, p. 906-915, Vol. 16, No. 6
1071-412X/09/$08.00+0     doi:10.1128/CVI.00413-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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