Development and application of species-specific cell-based bioassays to assess toxicity in green sea turtles

Abstract

Despite the detection of a wide range of contaminants in the blood of green turtle populations foraging in three locations of northern Queensland - Upstart Bay, Cleveland Bay and the Howick Group of Reefs, little is known about the effects of these contaminants on turtle health. Newly developed cell-based bioassays using green turtle primary cell cultures provide an ethical, reproducible, and high-throughput method for assessing the risk of chemical exposure sea turtles. In this project, the toxicity of six priority metals (Mn, Co, Mo, As, Sb, Cu) and blood extracts from foraging turtles were tested in two bioassays adapted to green turtle primary skin and liver cells. Cytotoxicity of metals and blood extracts was measured in primary skin fibroblast cells using a resazurin assay. Glutathione-S-transferase (GST) activity was measured in primary skin fibroblasts and primary liver epithelial cells following exposure to metals and blood extracts. Arsenic, molybdenum, cobalt and copper were found to be cytotoxic to green turtle skin cells. Only manganese, cobalt and copper were found to alter GST activity, predominantly in skin cells, indicating a higher sensitivity of green turtle skin cells compared to liver cells. Effect concentrations of metals in both bioassays were above concentrations found in turtle blood. Turtle blood extracts from the three foraging grounds showed differences in cytotoxicity and GST activity. In both assays, blood extracts of turtles from Upstart Bay were the most toxic, followed by those from Cleveland Bay, then the Howick Reefs, suggesting turtles from Upstart Bay and Cleveland Bay may be at risk from current concentrations of organic contaminants. This study demonstrates that species-specific cell-based bioassays can be used effectively to assess chemical risk in sea turtles and their foraging grounds, and could be applied to assess chemical risk in other marine wildlife.

Keywords: Exposure; Metals; Organics; QuEChERS; Sea turtle; in vitro bioassay.