Marine shellfish exposed to the microalgae Karenia selliformis can accumulate gymnodimines (GYM). Shellfish samples collected from Beihai City in Guangxi Autonomous Region, and Ningde City in Fujian Province, in the South China Sea, as well as mussels Mytilus galloprovincialis fed on K. selliformis under laboratory conditions were analyzed. Gymnodimines and various fatty acid ester metabolites were detected in the clam Antigona lamellaris and pen shell Atrina pectinata, while no esters were found in the oyster Crassostrea sp. and the gastropod Batillaria zonalis despite positive detection of free GYM in both species. When present, the predominant acyl esters observed were 18:0-GYM-A and 20:1-GYM-A. Under laboratory conditions GYM-A was accumulated and metabolized to fatty acid esters in mussels exposed to K. selliformis, with 16:0-GYM-A and 20:1-GYM-A as the major variants. A novel compound with the same accurate mass as GYM-A and its 16:0 fatty acid ester were observed in the experimental mussels but was not present in the microalgal strain to which mussels were exposed. No significant differences of reactive oxygen species (ROS) levels and antioxidant enzymes were found between mussels fed on K. selliformis or GYM-free microalgae Isochrysis galbana. This suggests the accumulation of GYM and its metabolites does not significantly impact the physiological status of mussels. While it is currently not proven that GYM affects human health, risk assessments should consider the presence of GYM esters in naturally contaminated shellfish as part of exposure analysis. Please note that this is an author-produced PDF of an article accepted for publication following peer review. The definitive publisher-authenticated version is available on the publisher Web site.
In recent years, detection of trace amounts of dissolved lipophilic phycotoxins in coastal waters has been possible using solid phase adsorption toxin tracking (SPATT) samplers. To explore the contribution of dissolved diarrhetic shellfish toxins (DST) to the accumulation of toxins by cultivated bivalves, mussels (Mytilus galloprovincialis) were exposed to different concentrations of purified okadaic acid (OA) and dinophysistoxin-1 (DTX1) in filtered (0.45 µm) seawater for 96 h. Accumulation and esterification of DST by mussels under different experimental conditions, including with and without the addition of the food microalga Isochrysis galbana, and with the addition of different size-fractions of suspended particulate matter (SPM) (<75 µm, 75–150 µm, 150–250 µm) were compared. Results showed that mussels accumulated similar amounts of OA and DTX1 from seawater with or without food microalgae present, and slightly lower amounts when SPM particles were added. Mussels preferentially accumulated OA over DTX1 in all treatments. The efficiency of the mussel’s accumulation of OA and DTX1 from seawater spiked with low concentrations of toxins was higher than that in seawater with high toxin levels. A large proportion of OA (86–94%) and DTX1 (65–82%) was esterified to DTX3 by mussels in all treatments. The proportion of I. galbana cells cleared by mussels was markedly inhibited by dissolved OA and DTX1 (OA 9.2 µg L−1, DTX1 13.2 µg L−1) in seawater. Distribution of total OA and DTX1 accumulated in the mussel tissues ranked in all treatments as follows: digestive gland > gills > mantle > residual tissues. However, the percentage of total DST in the digestive gland of mussels in filtered seawater (67%) was higher than with the addition of SPM particles (75–150 µm) (51%), whereas the gills showed the opposite trend in filtered seawater with (27%) and without (14.4%) SPM particles. Results presented here will improve our understanding of the mechanisms of DST accumulation by bivalves in marine aquaculture environments.
A number of new C-11 hydroxyl metabolites (so-called M-toxins) of paralytic shellfish toxins (PSTs) have been discovered in contaminated shellfish, and trace amounts have also been detected in some strains of PST-producing microalgae. To investigate the chemical conversion and stability of M-toxins, mussel extracts were purified with solid-phase extraction cartridges (Oasis HLB) and Biogel P-2 resin columns and four partially purified M-toxin fractions were stored at different temperatures (−20, 4, and 20 °C) and pH values (3, 4, and 5). The concentrations and profiles of M-toxins in these fractions were analyzed using liquid chromatography coupled with tandem mass spectrometry for 27 weeks. Results further confirmed the chemical conversion pathway M1 → M3 → M5 and determined for the first time two new transformation pathways: M2 → M4 → M6 and neosaxitoxin (NEO) → M10. The half-lives of M1, M2, M4, and M10 were calculated using a first-order degradation kinetics model, which indicated that the degradation of all M-toxins was dependent upon the temperature and pH, increasing with rising temperature and pH. In comparison to M4 and M10, M1 was more sensitive to the temperature, followed by M2. Results suggest that M-toxins should be maintained at a low temperature (−20 °C) and low pH (3) for their prolonged storage. M-toxins were less stable than all of the common analogues of PSTs, which may be beneficial for shellfish to achieve rapid detoxification through transformation of PSTs to M-toxins. These new findings are of significance because they enable further understanding of the metabolism of PSTs and their detoxification mechanisms in contaminated shellfish.
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