In the current study, we compared protein profiles in gills of Perna viridis after exposure to Prorocentrum lima, a dinoflagellate producing DSP toxins, and identified the differential abundances of protein spots using 2D-electrophoresis. After exposure to P. lima, the level of okadaic acid (a main component of DSP toxins) in gills of P. viridis significantly increased at 6 h, but mussels were all apparently healthy without death. Among the 28 identified protein spots by MALDI TOF/TOF-MS, 12 proteins were up-regulated and 16 were down-regulated in the P. lima-exposed mussels. These identified proteins were involved in various biological activities, such as metabolism, cytoskeleton, signal transduction, response to oxidative stress and detoxification. Taken together, our results indicated that the presence of P. lima caused DSP toxins accumulation in mussel gill, and might consequently induce cytoskeletonal disorganization, oxidative stress, a dysfunction in metabolism and ubiquitination/proteasome activity.

We reported previously that okadaic acid (OA) and dinophysistoxin-1 (DTX1) were responsible for diarrhetic shellfish poisoning (DSP) incidents due to consuming cultivated mussels (Mytilus galloprovincialis) in coastal cities near the East China Sea in May 2011. Pectenotoxin-2 (PTX2) and its seco acids were also present in these mussels. Causative species of microalgae were not identified because detailed information on the location of the contaminated shellfish was not recorded. In order to explore potential causes for these poisoning events, the lipophilic toxin profiles in picked cells of Dinophysis and in shellfish samples collected from two mariculture zones in the East China Sea were analyzed in the present study. Single-cell isolates (100 cells total for each location) of Dinophysis were collected from the aquaculture zones of Gouqi Island (Ningbo City, Zhejiang Province) and Qingchuan Bay (Ningde City, Fujian Province) in July and September 2013, respectively, for lipophilic toxin profiling. Shellfish samples collected over the course of a year from the Gouqi Island aquaculture zone and mussels (M. galloprovincialis) collected four times from the Qingchuan Bay aquaculture zone were tested for lipophilic toxins by LC-MS/MS. The Dinophysis cells isolated from both sampling sites were identified under the light microscope as Dinophysis caudata. Average quota of PTX2, the predominant toxin in D. caudata isolated from the coastal waters of Gouqi Island and Qingchuan Bay, was 0.58 and 2.8 pg/cell, respectively. Only trace amounts of OA and DTX1 were detected in D. caudata. PTX2, PTX2sa, 7-epi-PTX2sa, OA, and/or DTX1 were found in samples of mussels (M. galloprovincialis and Mytilus coruscus) collected in the Gouqi Island aquaculture zone from the end of May to the beginning of July 2013. PTX2, PTX2sa, and 7-epi-PTX2sa were also detected in oyster (Crassostrea gigas) during that period, but almost no OA and DTX1 were present. Gymnodimine (GYM) was detected in almost all mussel (M. coruscus) samples, with the highest levels occurring in winter. Trace amounts of pectenotoxins (PTXs) and OAs were also found in mussels (M. galloprovincialis) collected from Qingchuan Bay. D. caudata is suggested as an important source of PTXs in shellfish cultivated in the East China Sea. This is the first report of toxin profiles for single-cell isolates of Dinophysis in the East China Sea.

This constitutes the first study to report on the reduction in toxicity of the dinoflagellate algal toxin okadaic acid after novel pulsed light (PL) treatments where ecotoxicological assessment was performed using a miniaturised format of the conventional in vivo freshwater crustacean Daphnia sp. acute toxicity test. Bivalves accumulate this toxin, which can then enter the human food chain causing deleterious health effects such as diarrhetic shellfish poisoning. This miniaturised toxicological bioassay used substantially less sample volume and chemical reagents. Findings revealed a 24-h EC₅₀ of 25.87 μg/L for PL-treated okadaic acid at a UV dose of 12.98 μJ/cm² compared to a 24-h EC₅₀ of 1.68 μg/L for the untreated okadaic acid control, suggesting a 15-fold reduction in toxicity to Daphnia pulex. The bioassay was validated in this study and correlated well with the “classic” ISO format (r = 0.98) using the traditional reference chemical potassium dichromate (K₂Cr₂O₇). Reduction by up to 65% in PL-treated okadaic acid concentration was confirmed by LC-MS/MS analysis. Findings from this study have positive ecological, societal and enterprise implications, such as the development of PL technology for the prevention or reduce algal contamination of fisheries and aquaculture industries.