Comparison of Species Specific Antisera and Cross-reactive Antisera in an Elisa for the Determination of Serum Antibody Titers against Erysipelothrix rhusiopathiae in Delphinapterus leucas and Lagenorhyncus obliquidens
Rhonda A. Patterson; John C. Jones; Bobby L. Middlebrooks
University of Southern Mississippi, Department of Biological Sciences,
Hattiesburg, MS
Abstract
Erysipelas is considered one of the most problematic of diseases seen at
facilities that house cetaceans. Cetacean erysipelas is usually manifested in one of two forms: a
dermatological form, which appears as diamond- or square-shaped lesions mainly seen on the
anterior dorsal area of the skin, or an acute septicemic form. In order to provide a diagnostic
tool for routinely monitoring antibody titers against Erysipelothrix rhusiopathiae a
biotin-avidin amplified enzyme linked immunosorbent assay (ELISA) was developed. This assay
employs a "double indirect" indicator antibody system that includes biotin labeled
rabbit anti-Tursiops truncatus immunoglobuhn antiserum (produced against ammonium sulfate
precipitated immunoglobulins) followed by avidin-labeled alkaline phosphatase. In initial
determinations of anti-Erysipelothrix antibody levels with serum samples from
Delphinapterus leucas and Lagenorhyncus obliquidens, biotin labeled rabbit
anti-T. Truncatus immunoglobuhn anti which cross-reacts with immunoglobulins from those
species, was employed. The degree of cross-reactivity of the antisera was a variable in the assay
that could not be appropriately defined, so species specific antisera were produced and biotin
labeled. In order to determine if the use of species specific antisera would result in greater
assay sensitivity it was necessary to test serum samples from D. leucas (provided by Naval
Research and Development, San Diego, CA and the John G Shedd Aquarium, Chicago, II,) and L.
obliquidens (provided by the John G. Shedd Aquarium) under both the initial assay conditions
and with the appropriate species specific antisera. The results of this study indicated a
relative increase in recorded titers and the number of animals showing titers against E.
rhusiopathiae when the species specific antisera was utilized compared with the titers
determined when anti-T. Truncatus was utilized.
Acknowledgements
This work was supported by a grant from the John G Shedd Aquarium. The
authors wish to thank Dr. J. Boehm (John G. Shedd Aquarium) and Dr. S. Ridgway, Dr. B. Van Bonn,
and Dr. E. Jensen (NRAD) for their generous donation of samples for this study.