Project Summary/Abstract: Aquatic organisms are powerful biomedical models used in laboratories around the world. These animals are maintained mostly as live organisms, with the associated expense and risk. As thousands of new lines of mutants and transgenic organisms are generated, maintaining populations of each becomes prohibitive for stock centers. To address these problems, cryopreservation has become a necessity for the preservation of genetic resources. Cryopreservation must be linked to development of germplasm repositories where genetic resources are protected in a reproducible, standardized way. A well developed and maintained repository can reduce the cost of maintaining and recovering lines, reduce duplication of effort, and reduce the costs required to recreate lines. We are working to develop appropriate processes and repository technology for amphibians (i.e., Ambystoma and Xenopus) and invertebrates (i.e., Aplysia). Our current parent award is intended to provide access to community-level approaches for reproducibility, training, and standardization through a centralized resource Hub and a repository network that can address aquatic biomedical model repository needs. Through R24 funding the Hub is establishing capabilities to process germplasm samples and develop pathways for repository development. Currently, animals used for the project are sent from stock centers and other laboratories to the AGGRC. Our reliance on outside sources has become a substantial problem in ensuring the availability of adequate numbers of animals for replicated studies and outreach activities. Supplemental funds are requested herein to enable us to spawn and grow animals in-house. This will greatly advance the objectives of the parent award by increasing research productivity and outreach capability. Also, the risks associated with animal shipping, particularly in extreme weather conditions will be diminished. This work can be completed within the current project year (entering Year 3), and it does not overlap with work funded in the parent award or through previous supplemental funding. The specific use of this request is to provide in-house production live amphibians cultured from spawning through adulthood, to provide a timely source of broodstock to support the activities of the parent award in research, teaching, training, collaboration, and outreach.