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/100g. Oysters were toxic by mouse bioassay at levels >or=20 MU/100g 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.
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ABSTRACT: Harmful algal blooms can cause mass mortalities of top predators such as fish, marine mammals and seabirds but the food web transfer from toxic phytoplankton to these organisms has not been fully elucidated. Macrobenthic invertebrates in coastal waters, including bivalve suspension- and deposit-feeders, carnivorous gastropods, deposit-feeding amphipods and polychaetes, are a major food source for a wide variety of predators and can thus play a critical role in the trophic transfer of algal toxins to higher trophic levels. The objective of this study was to investigate toxin accumulation in transplanted juvenile hard clams, Mercenaria mercenaria, a species naturally occurring in the region, and in various macrobenthic functional groups from Florida coastal waters during a natural bloom of the dinoflagellate, Karenia brevis, a producer of brevetoxins. Bloom concentrations in the water column ranged from 100 to 1200 cells ml−1 over the course of the experiment. This study revealed that these lipophilic toxins can be rapidly accumulated by both suspension- and deposit-feeding benthos, especially bivalve molluscs [1.9–2.8 μg PbTx-3 eq (g wet weight)−1]. Transplanted M. mercenaria rapidly accumulated toxins from the water column attaining ∼0.5 μg PbTx-3 eq (g wet tissue)−1 after only 4 h-exposure to the K. brevis bloom and a maximum value of 1.5 ± 0.2 μg PbTx-3 eq (g wet tissue)−1 after 72 h. Relatively high brevetoxin concentrations were also measured in co-occurring benthic carnivorous gastropods [1–2.6 μg PbTx-3 eq (g wet weight, WW)−1]. Mean toxin concentrations in polychaetes and crustaceans varied in the range ∼0.04–0.2 μg PbTx-3 eq (g WW)−1 over the study period, and thus were typically lower than in molluscs. This study demonstrated in situ toxin accumulation by benthic primary and secondary consumers during a natural Florida red tide. Accumulation by primary consumers may be highly variable in space and time (as shown in bivalves from the natural benthic community) and among taxonomic groups. Toxin transfer further up the food web will thus depend on the toxin level accumulated in prey, the number of pathways from which the predator may accumulate toxins and on possible biological magnification of lipophilic toxins. Overall, this study revealed qualitatively and quantitatively that benthic consumers of a number of taxa can serve as vectors for transporting brevetoxins within the food web.Harmful Algae 04/2012; 16:27–34. DOI:10.1016/j.hal.2012.01.001 · 3.34 Impact Factor
European Neuropsychopharmacology 09/2011; 21. DOI:10.1016/S0924-977X(11)71024-0 · 5.40 Impact Factor
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ABSTRACT: Mouse bioassays have been a mainstay for detecting harmful concentrations of marine algal toxins in shellfish for over 70 years. Routine monitoring involves intraperitoneal injection of shellfish extracts into mice; shellfish contaminated with algal toxins are thus identified by mortality in exposed mice. With the advent of alternative test methods to detect and quantify specific algal toxins has come increasing criticism of enduring use of mouse bioassays for shellfish safety testing. However, the complete replacement of shellfish safety mouse bioassays by chemical, antibody-based, and functional assays has been and will continue to be a gradual process for various reasons, including skills availability and instrument costs for chromatography-based toxin monitoring. Mouse bioassays for shellfish safety testing do not comply with modern standards for laboratory animal welfare, specifically the requirement in published official methods for death as a test outcome. Mouse bioassays for algal biotoxins in shellfish, as well as fundamental algal toxin research endeavors using in vivo models, are amenable to revision and refinement from a humane endpoints perspective. Regulated hypothermia may be a useful and easily acquired nonlethal toxicological endpoint; objective determination of neuromuscular blockade may allow algal neurotoxin testing and research to enter the domain of humane endpoints evaluation. Relinquishing reliance on subjective test endpoints, including death, will likely also deliver collateral improvements in assay variability and sensitivity.Journal of AOAC International 03/2014; 97(2):356-72. DOI:10.5740/jaoacint.SGEStewart · 1.39 Impact Factor