The unifying goal of our work is to exploit the tremendous potential of the captive breeding and reintroduction program to recover the louisiana pinesnake

This federal Project Grant award, provided by the U.S. Department of the Interior Fish and Wildlife Service under the Endangered Species Recovery Implementation program (CFDA 15.657), supports a comprehensive conservation research initiative focused on recovering the endangered Louisiana Pinesnake.

The $232,668 award, granted to the Memphis Zoological Society, will fund several key activities over a 2-year period from June 2024 to May 2026. These include: improving cryopreservation and artificial insemination technologies to preserve genetic diversity; constructing genomic pedigrees for captive and reintroduced wild populations; detecting the presence of diseases that threaten wild Louisiana Pinesnakes; and testing effective release protocols to optimize the success of the captive breeding and reintroduction program. The project involves collaboration with several partner organizations, including other zoos, research institutions, and government agencies, to pool expertise and resources for this critical endangered species recovery effort.

The reintroduction program is an impactful conservation action for the pinesnake, having placed 408 ZOO-bred juveniles onto restored habitat in kisatchie national forest, louisiana. Young pinesnakes have been confirmed to be offspring of released individuals, indicative that a new population is taking hold in this unoccupied portion of the historic range. Despite early success, the pinesnake release program would benefit from several components to amplify its contribution to species recovery. These are: knowledge-based biosecurity protocols to improve captive and wild snake habitat welfare, increased reproductive rates through improved reproduction technologies, preserving genetic diversity by using genomic lineage data and reproduction technologies, scientific data to inform the best release protocol, and effective detection methods to guide the selection of future release sites. The memphis ZOO, fort worth ZOO, ellen trout ZOO, audubon ZOO, US army corps of engineers engineer research and development center, and university of illinois are pooling their expertise on a body of conservation research that will address all priority 1 and 2 actions in the draft recovery plan for the louisiana pinesnake.by improving cryopreservation and artificial insemination technologies developed by the memphis ZOO, we will both preserve in situ genetic diversity and increase captive genetic diversity. This will be accomplished by collecting and cryobanking semen from wild-caught louisiana pinesnakes and fertilizing captive females via artificial insemination to produce offspring carrying native genetic diversity which will capture this diversity and strengthen the genome of the captive population. We must base future management decisions for the captive population and in situ reintroduction program using actual genetic information. For the captive population, we will construct a multigenerational genomic pedigree and reveal a precise estimation of the degree of relatedness between every individual. For the reintroduced in-situ population, we will determine founder representation in natural recruitment. For wild louisiana pinesnakes, we will estimate the genomic diversity in the current and historical natural populations. We must protect louisiana pinesnake habitat, of which an important component is the disease risk to wild populations. Snake fungal disease and cryptosporidium are two known pathogens that threaten snake health. Both organisms are capable of decimating wild populations and causing local extinctions. Therefore, via DNA analysis of skin and rectal swabs, we will detect the presence of both organisms in pinesnakes and other snakes living in pinesnake habitat. This research will benefit state and federal agencies in developing strong release protocols that protect naturally occurring louisiana pinesnakes from disease. Furthermore, to prevent introduction of pathogens by captive bred releases into naturalhabitat, we will test all snakes in the captive population for the two diseases, and screen candidate snakes before they are approved for release. We must optimize the success of the reintroduction program by employing the best species and habitat specific release protocol. To do this will conduct an experimental release to discern spatial use and behaviors of ZOO-bred pinesnakes under different release procedures to determine which strategy yields the best results in terms of acclimation and survival.finally, practical and effective detection methods are critical in order to discover any currently undocumented wild populations, and for selecting appropriate unoccupied sites for expanded reintroduction programs. We will test and develop edna sampling methodology to apply on louisiana pinesnake habitat.