Evaluation of immunological techniques for host fish identification, and cryopreservation of embryos for conserving rare freshwater mussels

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1993-09-16
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Virginia Tech
Abstract

Glochidia (larvae) of freshwater mussels are obligate parasites which attach to and become encysted in the gills or fins of host fish species. The immune responses of the host fish to the parasite affects the susceptibility of the fish to glochidia of different mussels. The immune response provides an opportunity to identify which fish species are hosts. The number and variety of mussels in rivers and lakes has sharply declined since the last century due to various anthropogenic factors, and some mussels species are facing extinction. It is an urgent task to preserve these vanishing mussels, or extinction will be inevitable.

An attempt was made to develop an assay, using the immunological response to glochidia, to screen fish species for appropriate hosts. This would facilitate the production and rearing of juveniles. In order to design these assays, reagents such as anti-immunoglobulins which can react with antibodies from many different fish species have to be developed. This work was carried out to develop such reagents. Host and non-host fish were immunized with killed bacteria (Brucella abortus) to study their humoral immune response to an antigen. All fish were able to respond well, as measured by agglutination and Western Blot assays. Antibodies produced by the Brucella injections were used to stimulate anti-fish immunoglobulins in goats, and the antisera were tested for their ability to recognize immunoglobulins from different host fish species. The specificities of these reactions were compared to the reactivity of Protein A. Goat antisera were able to cross-react with different fish antibodies, but it was found that Protein A was a more suitable reagent. Protein A is seemingly suitable to identify the host-fish species and could be used as a reagent for the serological diagnosis of various fish diseases.

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