Source: KENT SEATECH CORP. submitted to
DEVELOPMENT OF LARGE-SCALE SPERM CRYOPRESERVATION TECHNIQUES FOR STRIPED BASS AQUACULTURE
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
0199523
Grant No.
2004-33610-14314
Cumulative Award Amt.
(N/A)
Proposal No.
2004-00423
Multistate No.
(N/A)
Project Start Date
May 15, 2004
Project End Date
Jun 30, 2005
Grant Year
2004
Program Code
[8.7]- (N/A)
Recipient Organization
KENT SEATECH CORP.
11125 FLINTKOTE AVE., SUITE J
SAN DIEGO,CA 92121
Performing Department
(N/A)
Non Technical Summary
Hybrid striped bass aquaculture is the fourth largest form of U.S. finfish production U.S. producers and university scientists recently formed a National Breeding Program to develop superior strains. To take advantage of the new strains and use them in commercial production facilities, methods of reliable storage and distribution of broodstock gametes are needed. Cryopreservation of sperm offers an inexpensive method of storing and distributing the gametes of promising strains. Working with the LSU Dairy Improvement Center, we will develop techniques for freezing, storing, and distributing striped bass sperm that take advantage of large-scale dairy cryopreservation facilities which offer modern, standardized and cost-effective methods for cryopreservation of sperm. Development of large-scale, standardized technologies for cryopreservation of striped bass and white bass sperm will be extremely beneficial in the commercial culture of hybrid striped bass. Cost-effective methods for cryopreserving, storing, and transporting sperm from selectively bred broodstock will assist in providing rapid-growing and disease-resistant strains of fingerlings to U.S. producers for grow-out. Also, since the sperm will be available throughout the year, eggs from female broodfish could be fertilized at any time. This will facilitate production of fingerlings and stocking of grow-out facilities on a year-round basis, which will greatly enhance production efficiencies in both intensive and extensive culture systems nationwide.
Animal Health Component
100%
Research Effort Categories
Basic
(N/A)
Applied
100%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
30137131020100%
Knowledge Area
301 - Reproductive Performance of Animals;

Subject Of Investigation
3713 - Striped bass;

Field Of Science
1020 - Physiology;
Goals / Objectives
The overall goal of our Phase I SBIR research is to determine whether large-scale cryopreservation techniques such as those used in the dairy industry will be applicable in the aquaculture of hybrid striped bass. Studies will be conducted to develop optimal techniques for collecting, extending, and storing white bass and striped bass sperm. Also, we will determine the optimal requirements for automated, large-scale white bass and striped bass sperm cryopreservation, in cooperation with the Louisiana State University Agricultural Center and Genex Custom Collection, Inc., a farmers' cooperative that operates a modern and automated high-throughput cryopreservation facility in Baton Rouge, LA. The specific technical objectives of our Phase I SBIR research are: 1) to collect and evaluate characteristics of milt from white bass and striped bass broodstock, 2) to develop cold-storage and shipment methodologies for white bass and striped bass sperm, 3) to develop high-throughput cryopreservation techniques for white bass and striped bass sperm, and 4) to evaluate economic benefits to be derived from white bass and striped bass sperm cryopreservation.
Project Methods
Kent SeaTech and other private researchers, academic collaborators, and government scientists have established the National Breeding Program for the Hybrid Striped Bass Industry, a unique collective effort focused on the domestication and genetic improvement of striped bass and white bass. For this program to succeed, efficient methods of maintaining and distributing strains easily among the participating investigators will be required. Also, once genetically superior lines are developed, technologies will be required for safe storage and large-scale commercial distribution of gametes to producers throughout the U.S. One very promising method for large-scale distribution of gametes is through the use of cryopreservation technology. Sperm from selected male broodstock can be collected and cryopreserved and stored for long periods and then easily airshipped to users as needed. Although previous researchers have been able to develop sperm cryopreservation techniques for striped bass (e.g. Kerby (1983, Brown and Brown 2000, Jenkins-Keeran and Woods 2002) and several other fish species, there have been no practical applications of cryopreservation technology in the aquaculture industry to date. This is likely due to several inhibitory factors, including the need for costly specialized equipment, lack of technical expertise, and the inability to freeze and store the large quantities of male germplasn that would be needed for commercial production. Kent SeaTech is conducting SBIR Phase I research to develop efficient, large-scale sperm cryopreservation and storage technologies for use in the hybrid striped bass industry. The large-scale techniques we propose to develop are similar to those used in the dairy industry, where sperm cryopreservation has become an indispensable component in breeding and genetic improvement and has created an entirely new commercial industry. Large-scale cryopreservation technologies will reduce many of the risks and costs involved in maintaining broodstock and serve to accelerate selective breeding efforts, which should allow the hybrid striped bass industry to achieve more efficient production, reduce costs, and compete more effectively in the global seafood markets. The research is being conducted in cooperation with two outstanding academic scientists: Dr. Terrence R. Tiersch, a Professor at the Louisiana State University Agricultural Center, and Dr. Craig V. Sullivan, a Professor at North Carolina State University.

Progress 05/15/04 to 06/30/05

Outputs
The overall objective of this USDA SBIR research project was to determine the basic cryopreservation parameters and techniques that would be required in a large-scale, automated approach to cryopreservation of white bass and striped bass sperm for the hybrid striped bass culture industry. The specific technical objectives of our Phase I SBIR research were: 1) to collect and evaluate characteristics of milt from white bass and striped bass broodstock, 2) to develop cold-storage and shipment methodologies for white bass and striped bass sperm, 3) to develop high-throughput cryopreservation techniques for white bass and striped bass sperm, and 4) to evaluate economic benefits to be derived from white bass and striped bass sperm cryopreservation. We made excellent progress toward all of these objectives. Preliminary studies were conducted to evaluate a variety of techniques for collecting, extending, and storing sperm from these species. Using reagents and techniques developed from these studies, we were able to successfully cold-store white bass and striped bass sperm for 24-72 hours and maintain motilities that yielded fertilization rates that were often comparable to those attained with fresh semen. We then used these cold-storage techniques to ship samples and conduct experiments using an automated, large-scale white bass and striped bass sperm cryopreservation platform, in cooperation with the Louisiana State University Agricultural Center and Genex Custom Collection, Inc., a farmers' cooperative that operates a modern and automated high-throughput cryopreservation facility in Baton Rouge, LA. Successful fertilization of white bass eggs using cryopreserved sperm in small-scale and commercial-scale trials (the first ever conducted to our knowledge) provided excellent proof-of-principle for these new technologies. If we are able to expand upon our successful Phase I results and develop reliable and commercially-viable cyropreservation techniques during Phase II, U.S. culturists will have an important new production technology that could significantly enhance hybrid striped bass aquaculture.

Impacts
The advanced techniques for large-scale cryopreservation of white bass and striped bass sperm that we are developing will be of considerable assistance to private, federal, and academic hatcheries breeding these species. University and private researchers have joined together to form the National Breeding Program for the Genetic Improvement and Selective Breeding of the Hybrid Striped Bass Industry. The results from this research will help these scientists maintain and distribute the improved strains that are being developed for industry-wide use. The research also will assist the National Animal Germplasm Program (NAGP), a research group formed to preserve the genetic diversity of agriculturally important species. Samples of cryopreserved sperm from defined white bass and striped bass lines used in these studies, along with data on the quality of those sperm samples, is being deposited and stored at the NAGP repository. Also, successful development of our cryopreservation model involving a standardized central freezing center and efficient analysis of sperm quality will serve as a gateway for the cryopreservation and deposit of sperm samples of other aquatic species for preservation of genetic diversity with the NAGP.

Publications

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