Monitoring of brevetoxins in the Karenia brevis bloom-exposed Eastern oyster ( Crassostrea virginica)

Abstract

Brevetoxin uptake and elimination were examined in Eastern oyster ( Crassostrea virginica) exposed to recurring blooms of the marine alga Karenia brevis in Sarasota Bay, FL, over a three-year period. Brevetoxins were monitored by in vitro assays (ELISA, cytotoxicity assay, and receptor binding assay) and LC-MS, with in vivo toxicity of shellfish extracts assessed by the traditional mouse bioassay. Measurements by all methods reflected well the progression and magnitude of the blooms. Highest levels recorded by mouse bioassay at bloom peak were 157 MU/100 g. Oysters were toxic by mouse bioassay at levels ≥20 MU/100 g for up to two weeks after bloom dissipation, whereas brevetoxins were measurable by in vitro assays and LC-MS for several months afterwards. For the structure-based methods, summed values for the principal brevetoxin metabolites of PbTx-2 (cysteine and cysteine sulfoxide conjugates), as determined by LC-MS, were highly correlated ( r 2 = 0.90) with composite toxin measurements by ELISA. ELISA and LC-MS values also correlated well ( r 2 = 0.74 and 0.73, respectively) with those of mouse bioassay. Pharmacology-based cytotoxicity and receptor binding assays did not correlate as well ( r 2 = 0.65), and were weakly correlated with mouse bioassay ( r 2 = 0.48 and 0.50, respectively). ELISA and LC-MS methods offer rapid screening and confirmation, respectively, of brevetoxin contamination in the oyster, and are excellent alternatives to mouse bioassay for assessing oyster toxicity following K. brevis blooms.