Progress 10/01/20 to 09/30/21
Outputs
Target Audience:Target audiences are members of the scientific community, aquaculture industry members, country extension agents, and administration agencies involvingwith aquaculture and fishery management, natural resourcebiodiversity, and genetic resourceconservation. Changes/Problems:No significant changes will be made. What opportunities for training and professional development has the project provided?One postdoctoral scientist, one graduate student, and four undergraduate students participated the research activities and got trained in this program within this reporting period. Training activities include scientific research activities, such as procedure demonstration and laboratory techniques, and extension activities, such as on-site visit and extension publications. Professional development activities include project discussion, supervisory special course, online-course, and participation of academic conferences. How have the results been disseminated to communities of interest?Dissemination of research updates were made through onsite demonstration, field trip visit, extension publications, graduate student thesis, and peer-reviewed journal publications. What do you plan to do during the next reporting period to accomplish the goals?During next reporting period, the planned research activities in the proposal will be continued to achieve project goal and objectives.
Impacts
What was accomplished under these goals?
A cost-effective and portable freezing system for germplasm cryopreservation of fish and shellfish was developed. A wide range of cooling rates (1.5 to 32.1 °C/min) were achieved by combination of positioning samples at 2 or 5 cm above liquid nitrogen surface, nitrogen vapor temperature of -150 or -180 °C, and aeration time of 0, 1, 2 min, or continuity. Compared to a programmable freezer, the aeration freezing system generated cooling profiles with significantly higher temperature for ice nucleation initiation. Cryopreservation of sperm of eastern oysters was conduction by use of the aerated freezing system and a computer programmable freezer. No difference in post-thaw sperm motility were found between samples cooling in the aeration freezing system or a programable freezer. Cooling rates of 10 or 15 °C/min from 4 °C to - 80°C yielded the highest post-thaw sperm motility regardless of the cooling devices. The aeration freezing system we developed has potential for high-throughput sample processing, and the design allows easy construction by multiple users. A sperm repository of the broodstock, including a total of 102 male oysters from the 17 collection sites, was established to support the breeding project, which was initiated in 2019 to support the fast-growing oyster aquaculture industry in the Gulf of Mexico. Sperm collection was accomplished by strip spawn, and fresh sperm production, motility, and fertility were recorded for quality analysis. Cryopreserved sperm samples were sorted, labelled, archived, and stored in liquid nitrogen for future use. Post-thaw motility and plasm membrane integrity were recorded as post-thaw quality parameters. Overall, this study demonstrated sperm sample collection, processing, cryopreservation, and a data management plan involved in the establishment of the sperm repository. The streamlined procedure can serve as a template for construction of oyster germplasm repositories for breeding programs. A review was conducted on the status and summary of larval cryopreservation in aquaculture mollusks. A total of 25 publications were identified addressing 11 molluscan bivalve species. Most studies were conducted on trochophores and D-larvae, a few studies were on umbo larvae, and no study was found on pediveliger. Based on the post-thaw viability, there is no general conclusion about the best larval stage for cryopreservation. Generally, most publications focused on developing or improving cryopreservation protocols through exploring one or more vital factors of the cryopreservation procedure. Evaluation of post-thaw viability was reported over a wide range of parameters with no standard criteria, for example motile post-thaw larvae were considered alive but no definition of swimming velocity. A sharp decrease of survivals was often reported after a period of culture although immediate post-thaw survivals were reported high in several studies. Overall, this review lay foundation for promoting molluscan larval cryopreservation technology for germplasm repository establishment and commercial seed production.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Yang H, Huo Y, Yee JC, Rikard S, Walton W, and Saillant E. 2021. Sperm repository for a breeding program of the Eastern oyster Crassostrea virginica: Sample collection, processing, cryopreservation, and data management plan. Animals, 11(10), 2836. https://doi.org/10.3390/ani11102836
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Huo Y, Yee JC and Yang H. 2021. Development of a cost effective and portable aeration freezing system for high-throughput sperm cryopreservation of eastern oysters Crassostrea virginica and other aquatic species. Aquaculture Research, https://doi.org/10.1111/are.15587
- Type:
Journal Articles
Status:
Published
Year Published:
2022
Citation:
Yang H and Huo Y. 2022. Review of molluscan larval cryopreservation and potential application on germplasm repository establishment and commercial seed production. Aquaculture, 547, 737491. https://doi.org/10.1016/j.aquaculture.2021.737491
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Progress 02/13/20 to 09/30/20
Outputs
Target Audience:Target audiences are members of the scientific community, aquaculture industry farmers, country extension agents, and administration agencies involvingwith aquaculture and fishery management, natural resourcebiodiversity, and genetic resourceconservation. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?One postdoctoral scientist, one graduate student, and one volunteer participated the research activities and got trained in this program within this reporting period despite of the covid-19 pandemic. Training activities include scientific research activities, such as procedure demonstration and laboratory techniques, and extension activities, such as on-site visit and extension publications. Professional development activities include project discussion, supervisory special course, online-course, and participation of academic conferences. How have the results been disseminated to communities of interest?Dissemination of research updates were made through onsite demonstration, field trip visit, extension publications, graduate student thesis, and peer-reviewed journal publications. What do you plan to do during the next reporting period to accomplish the goals?During next reporting period, the planned research activities in the proposal will be continued to achieve project goal and objectives.
Impacts
What was accomplished under these goals?
A novel aeration system was developed to create continuous supply of nitrogen vapor during the cooling process and thus achieving varied cooling rates. This system, named as "aeration freezing system", can be used for cryopreservation in field as required cooling rates from 5 to 25°C/min. Parallel comparison of this novel system and a programmable freezer was conducted for oyster sperm cryopreservation, the post-thaw sperm and similar post-thaw sperm motility (30-50%) was obtained. A sperm packaging system for cryopreservation was set up for field use. To package sperm samples quickly enough within certain time for cryopreservation, a sample filling system was constructed, including a filling station, a vacuum pump, and a straw sealing machine. The "aeration freezing system" and sample packaging system were applied on sperm cryopreservation for an ongoing oyster breeding program in the Gulf region. A total of 3893 straws (each straw = 0.5 ml) of cryopreserved samples were produced. The "aeration freezing system" produced the optimized cooling rates of 10.21 to 15.74 °C/min.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Yang H and Tiersch TR. 2020. Concepts, History, Principles, and Application of Germplasm Cryopreservation Technology. The Electronic Data Information Source of UF/IFAS Extension https://edis.ifas.ufl.edu/fa223
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