Laboratory-Scale Manipulation of Oysters

	Texas Chapter of the American Fisheries Society

From the 2000 Joint Meeting of the Arkansas, Louisiana, and Texas Chapters of the American Fisheries Society held in Bossier City, Louisiana.

Laboratory-Scale Manipulation of Oysters
Buchanan, J. T., Department of Oceanography and Coastal Sciences, Louisiana State University, Baton Rouge, LA 70803
Paniagua, C. G. and T. R. Tiersch, Aquaculture Research Station, Louisiana State University Agricultural Center, Baton Rouge,
LA 70803
Cooper, R. K., Department of Veterinary Science, Louisiana State University, Baton Rouge, LA, 70803

The eastern oyster Crassostrea virginica provides an important national fishery. There is a need for research in disease resistance and microbial elimination in oysters, and gene transfer research can lead to advances in this area. Cryopreservation of oyster gametes and larvae are important as well. To pursue research in these areas, it was necessary to develop research-scale techniques for the holding of oysters and the culture of larvae. There are unique needs associated with culturing oysters in the laboratory. Along with consistent production of high quality gametes and larvae, consideration must be given to experimental replication, avoidance of contamination, and containment of genetically modified organisms. All of our work was done with artificial seawater in recirculating systems over 100 km from the nearest coastal area. We examined the effect of several variables on the production of gametes and larvae of the eastern oyster. First, we developed protocols for holding broodstock in the laboratory, acclimation to laboratory conditions, and collection of gametes. We developed methods to assay gamete quality and for cold storage of gametes. Second, we optimized methods for the small-scale production of oyster larvae by artificial fertilization by examining the effect of container volume, aeration, and artificial water source on larval survival. Using these techniques, we successfully transferred the gene for red-shifted green fluorescent protein (rsGFP) into sperm and observed expression of the gene in oyster larvae. We have also observed expression of this
gene after transfer into adult oysters and in embryos. These techniques for oyster husbandry in the laboratory have been used in experiments to cryopreserve eastern oyster sperm and trochophore larvae. A program such as this would be useful for the small-scale production and culture of shellfish larvae for a variety of experimental purposes.

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