Metabotyping of a Protected Non-Model Organism, Green Sea Turtle (Chelonia mydas), using 1H NMR Spectroscopy and Optimized Plasma Methods for Metabolomics

The metabolomic fingerprints of a protected sea turtle species have been investigated for the first time using a nuclear magnetic resonance (NMR)-based metabolomics approach. We emphasized method development of optimal plasma filtration conditions (filter type, washing techniques, extract stability) for green turtles and other organisms, while also using the National Institute of Standards and Technology (NIST) Standard Reference Material 1950 (SRM 1950) Metabolites in Human Plasma for quality control. We surveyed the blood plasma metabolomic fingerprints of Hawaiian green sea turtles representing a wide range of physiological conditions that include varying disease states, behavioral conditions, and locales. The turtles sampled were free-swimming (n=5 from Hualalai on the west coast of the island of Hawaii), basking (n=7 from Hualalai), free-swimming tumor-free (n=3 from Kapoho, east coast of Hawaii), or freeswimming afflicted with external tumors (n=5 from Kapoho) caused by the disease, fibropapillomatosis (FP). The metabolomic profiles and the specific metabolites that differed among individual turtles are discussed. This optimized tool and the annotated metabolic profiles will benefit future investigations into the behavioral and disease conditions of the green turtle.