The complexity of threats to amphibians associated with climate change, pollution, habitat loss, anddisease (e.g. chytrid fungus, caused by the batrachochytrim dendrobatidis ,BD, demand new,…

This Project Grant award from the U.S. Department of the Interior Fish and Wildlife Service under the Endangered Species Recovery Implementation program (CFDA 15.657) provides $548,021 to Revive & Restore, a non-profit conservation organization, to develop new biobanking methods to enhance recovery and management of threatened amphibian species, with a focus on the endangered Wyoming toad (Anaxyrus baxteri).

The key goals of the project are to: (1) cryopreserve and recover live amphibian embryos, (2) generate, cryopreserve, and recover embryonic cell cultures, (3) generate, cryopreserve, and recover adult somatic cell cultures from tissue biopsies, and (4) use population genomics to identify high-value lineages for biobanking and captive breeding. These advanced biobanking and genetic rescue techniques aim to safeguard genetic diversity, enable disease resistance research, and potentially reduce costs for amphibian recovery efforts. If successful, the methods developed could be directly applicable to other endangered Anaxyrus toad species.

The complexity of threats to amphibians associated with climate change, pollution, habitat loss, anddisease (e.g. chytrid fungus, caused by the batrachochytrim dendrobatidis ,BD, demand new, innovativeapproaches to recovery efforts. According to the iucn classifications of threatened and endangeredspecies, amphibians are the most threatened group of vertebrates, with 41 percent of speciesthreatened with extinction. For comparison, the second most threatened group is mammals with 27PERCENT of species threatened. People working to save native amphibians in the U.S. And its territorieshave witnessed entire breeding populations vanish in the span of their careers. Innovative approacheswill take time to develop and deploy, during which important adaptive genetic diversity will be lostthrough population decline and or genetic drift. Biobanking technologies could safeguard amphibiangenetic diversity for future conservation management, but the existing toolkit for amphibians isextremely limited. Our proposed project aims to develop new biobanking methods to enhance recoveryand management of threatened amphibians, starting with the wyoming toad, anaxyrus baxteri, is one ofnorth americas most endangered amphibian species. The wyoming toads recovery plan is the onlyamphibian recovery plan that lists cryopreservation (biobanking) coupled with reproductive technologiesas a priority 1 action: action 4.3.1.5. Pursue sperm cryopreservation and in vitro fertilization (IVF). Ourproject aims to fulfill this action by expanding capabilities beyond sperm and IVF. Our goals are to: (1)cryopreserve and recover live embryos that develop into new reproductively viable individuals (2)generate, cryopreserve, and recover embryonic cell cultures (3) generate, cryopreserve, and recoveradult somatic cell cultures from non-lethally collected tissue biopsies and (4) use population genomicsto identify the highest value lineages in the pedigree for immediate biobanking and captive breeding.together, these strategies will enhance present and future genetic management efforts for the wyomingtoad and lay the groundwork for additional priority 1 recovery actions, including: 4.3.1.1. Maintaingenetic diversity of captive population, 4.3.1.6. Continue to reduce disease in captive toads, 4.3.2.1.continue genetic research, 4.3.3.2. Minimize spread of BD in the field. The ability to preserve andrestore both male and female genomes for breeding, through the goals we have proposed here (liveembryos or cell lines), would further reduce resources and costs to achieve genetic management goals. The methods proposed here for the wyoming toad may be directly transferable to other endangeredanaxyrus species (such as the houston toad, arroyo toad, yosemite toad, or dixie valley toad) and, ifsuccessful, embryo cryopreservation specifically could be broadly applied to all listed amphibian species.with effective biobanking tools, the wyoming toad recovery team, and other amphibian recovery teamscould immediately revolutionize recovery strategies, slow further gene diversity loss, enable diseaseresistance research, and potentially reduce overall breeding program costs.