SREL Reprint #3617

Peripheral blood hematology, plasma biochemistry, and the optimization of an in vitro immune-based assay in the brown watersnake (Nerodia taxispilota)

David Lee Haskins1,2, M. Kyle Brown2, Kristina Meichner3, Tracey D. Tuberville2, and Robert M. Gogal, Jr.4

1Interdisciplinary Toxicology Program, University of Georgia, Athens, Georgia, USA
2University of Georgia, Savannah River Ecology Laboratory, Aiken, South Carolina, USA
3Department of Pathology, University of Georgia College of Veterinary Medicine, Athens, Georgia, USA
4Department of Biosciences and Diagnostic Imaging, College of Veterinary Medicine,
University of Georgia, Athens, Georgia, Georgia, USA

Abstract: Reptiles represent a phylogenetic lineage that provides a unique link between ectothermic anamniotes and endothermic amniotes. Compared to mammalian and avian species, our understanding of the reptilian immune system is greatly lacking. This gap in knowledge is largely due to an absence of established immune-based assays or specific reagents for these species. In the present study, brown watersnakes (Nerodia taxispilota) were live-captured in the wild, sexed, weighed, measured, bled via the caudal vein, and released. At 24 hr post-collection, peripheral blood leukocytes were enriched and evaluated with an established mammalian in-vitro lymphocyte proliferation assay. Snake peripheral blood leukocyte enrichment yielded >90% lymphocytes with viabilities averaging 81.5%. Baseline physiologic data for N. taxispilota, including hematology and total solids, leukocyte differentials, cell recovery, and plasma biochemistry, were also collected. Cells cultured with Concanavalin A exhibited significantly increased proliferation at both 72 and 96 hr. These preliminary results show that enriched peripheral blood from wild-caught N. taxispilota provides a sufficient yield of leukocytes that can be cultured and functionally evaluated using a standard mammalian in-vitro immune-based assay.

Keywords: Snake; peripheral blood collection; hematology; lymphocyte separation; mitogen; proliferation

SREL Reprint #3617

Haskins, D. L., M. K. Brown, K. Meichner, T. D. Tuberville, and R. M. Gogal, Jr. 2021. Peripheral blood hematology, plasma biochemistry, and the optimization of an in vitro immune-based assay in the brown watersnake (Nerodia taxispilota). Journal of Immunoassay and Immunochemistry 42(1): 4-18.

This information was provided by the University of Georgia's Savannah River Ecology Laboratory (srel.uga.edu).