Development of an indirect immunofluorescence technique for the diagnosis of toxoplasmosis in bottlenose dolphins.
The diagnosis of toxoplasmosis is often complicated by the lack of specific clinical symptoms or postmortem features, in humans and other animals. The only diagnostic test described so far for the serological diagnosis of Toxoplasma gondii in marine mammals is the modified agglutination test (Dubey et al., Am J Vet Res 48(8):1239-1243, 1987). The development of more sensible and specific immunological techniques requires specific antibodies, which are currently unavailable in the scientific market. Indirect immunofluorescence (IIF) is one of the most widely used methods for the diagnosis of toxoplasmosis in humans (Auer et al., Parasitol Res 12:965-970, 2000). In order to develop and apply this technique to the bottlenose dolphin (Tursiops truncatus), immunoglobulins were firstly purified using ion-exchange chromatography. The purified immunoglobulins were then injected in New Zealand rabbits in order to obtain polyclonal antibodies. These antisera were validated by the IIF technique, using as controls serum samples of dolphins infected by Toxoplasma. The results were visualized using antirabbit IgG labeled with fluorescein. This newly developed and specific serological assay was then tested with the dolphin collection of Loro Parque, Tenerife, Spain (group I), and L'Oceanogràfic of Valencia, Spain (group II). The obtained results in this study showed that none of the dolphins from group 1 were infected by T. gondii and two animals were positive in group 2. Furthermore, we conclude that this study has produced antibodies with high specificity against dolphin immunoglobulins and an IIF method which may be used as immunological diagnostic tools, especially for the serological diagnosis of toxoplasmosis.
University Institute of Tropical Diseases and Public Health of the Canary Islands, University of La Laguna, Av. Astrofísico F. Sánchez s/n, 38271, La Laguna, Tenerife, Spain, mjosebernalg@yahoo.es.
María José
MJ
Bernal-Guadarrama
Joan
J
Salichs
Javier
J
Almunia
Daniel
D
García-Parraga
Nuhacet
N
Fernández-Gallardo
María Ángeles
MÁ
Santana-Morales
Víctor
V
Pacheco
Raquel N
RN
Afonso-Lehmann
Daniel
D
Déniz
Jacob
J
Lorenzo-Morales
Basilio
B
Valladares
Enrique
E
Martínez-Carretero
D000372:Agglutination Tests; D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood*; D050681:Bottle-Nosed Dolphin / Q000276:immunology / Q000469:parasitology*; D002852:Chromatography, Ion Exchange; D019084:Fluorescent Antibody Technique, Indirect; D011817:Rabbits; D013030:Spain; D014122:Toxoplasma / Q000276:immunology*; D014124:Toxoplasmosis, Animal / Q000175:diagnosis* / Q000276:immunology
D000913:Antibodies, Protozoan