Occurrence and Exposure Assessment of Lipophilic Shellfish Toxins in the Zhejiang Province, China.

Although lipophilic shellfish toxins (LSTs) pose a significant threat to the health of seafood consumers, their systematic investigation and risk assessment remain scarce. The goals of this study were as follows: (1) analyze LST levels in commercially available shellfish in Zhejiang province, China, and determine factors influencing LST distribution; (2) assess the acute dietary risk of exposure to LSTs for local consumers during the red tide period; (3) explore potential health risks of LSTs in humans; and (4) study the acute risks of simultaneous dietary exposure to LSTs and paralytic shellfish toxins (PSTs). A total of 546 shellfish samples were collected. LSTs were detected in 89 samples (16.3%) at concentrations below the regulatory limits. Mussels were the main shellfish species contaminated with LSTs. Spatial variations were observed in the yessotoxin group. Acute exposure to LSTs based on multiple scenarios was low. The minimum tolerable exposure durations for LSTs calculated using the mean and the 95th percentile of consumption data were 19.7 and 4.9 years, respectively. Our findings showed that Zhejiang province residents are at a low risk of combined exposure to LSTs and PSTs; however, the risk may be higher for children under 6 years of age in the extreme scenario.

Contamination Status and Risk Assessment of Paralytic Shellfish Toxins in Shellfish along the Coastal Areas of China.

Paralytic shellfish toxins (PSTs) are widely distributed in shellfish along the coast of China, causing a serious threat to consumer health; however, there is still a lack of large-scale systematic investigations and risk assessments. Herein, 641 shellfish samples were collected from March to November 2020, and the PSTs' toxicity was detected via liquid chromatography-tandem mass spectrometry. Furthermore, the contamination status and potential dietary risks of PSTs were discussed. PSTs were detected in 241 shellfish samples with a detection rate of 37.60%. The average PST toxicities in mussels and ark shells were considerably higher than those in other shellfish. The PSTs mainly included N-sulfonylcarbamoyl toxins (class C) and carbamoyl toxins (class GTX), and the highest PST toxicity was 546.09 µg STX eq. kg-1. The PST toxicity in spring was significantly higher than those in summer and autumn (p < 0.05). Hebei Province had the highest average PST toxicity in spring. An acute exposure assessment showed that consumers in Hebei Province had a higher dietary risk, with mussels posing a significantly higher dietary risk to consumers. This research provides reference for the green and sustainable development of the shellfish industry and the establishment of a shellfish toxin prevention and control system.

Detection of paralytic shellfish toxins by near-infrared spectroscopy based on a near-Bayesian SVM classifier with unequal misclassification costs.

BACKGROUND: Paralytic shellfish poisoning caused by human consumption of shellfish fed on toxic algae is a public health hazard. It is essential to implement shellfish monitoring programs to minimize the possibility of shellfish contaminated by paralytic shellfish toxins (PST) reaching the marketplace. RESULTS: This paper proposes a rapid detection method for PST in mussels using near-infrared spectroscopy (NIRS) technology. Spectral data in the wavelength range of 950-1700 nm for PST-contaminated and non-contaminated mussel samples were used to build the detection model. Near-Bayesian support vector machines (NBSVM) with unequal misclassification costs (u-NBSVM) were applied to solve a classification problem arising from the fact that the quantity of non-contaminated mussels was far less than that of PST-contaminated mussels in practice. The u-NBSVM model performed adequately on imbalanced datasets by combining unequal misclassification costs and decision boundary shifts. The detection performance of the u-NBSVM did not decline as the number of PST samples decreased due to adjustments to the misclassification costs. When the number of PST samples was 20, the G-mean and accuracy reached 0.9898 and 0.9944, respectively. CONCLUSION: Compared with the traditional support vector machines (SVMs) and the NBSVM, the u-NBSVM model achieved better detection performance. The results of this study indicate that NIRS technology combined with the u-NBSVM model can be used for rapid and non-destructive PST detection in mussels. © 2023 Society of Chemical Industry.

Occurrence and risk assessment of okadaic acid, dinophysistoxin-1, dinophysistoxin-2, and dinophysistoxin-3 in seafood from South Korea.

The okadaic acid (OA)-group toxins, including OA, dinophysistoxin-1 (DTX1), dinophysistoxin-2 (DTX2), and dinophysistoxin-3 (DTX3), cause diarrheic shellfish poisoning in humans. To manage OA-group toxins more strictly, Korean regulations were recently revised to consider OA, DTX1, DTX2, and DTX3 combined. Thus, our study characterized the occurrence of OA, DTX1, DTX2, and DTX3 in seafood distributed across South Korea, and a risk assessment of seafood consumption was conducted. Two hundred and seventeen samples from 16 bivalve and 7 non-bivalve species collected from three representative coastal areas in 2021 were analyzed via liquid chromatography-tandem mass spectrometry. OA, DTX1, and DTX3 were detected in 2.3%, 4.1%, and 9.2% of the examined samples, with positive mean levels of 11.3, 16.4, and 40.9 µg/kg, respectively. DTX2 was not detected in any of the samples. At least one OA-group toxin was detected in the bivalve samples, including blood clams, pan shells, hard clams, mussels, and scallops, whereas none were detected in non-bivalves. The estimated acute exposure to OA-group toxins through the intake of seafood in the Korean population and consumer groups was low, ranging from 24.7 to 74.5% of the recommended acute reference dose (ARfD) of 0.33 µg OA equivalents/kg body weight. However, for the scallop consumers aged 7-12 years, acute exposure to OA-group toxins exceeded the ARfD, indicating a possible health risk. These results suggest that including DTX3 in the new regulatory limits is appropriate to protect Korean seafood consumers from exposure to OA-group toxins.

An integrated biological effects assessment of the discharge water into the Sunndalsfjord from an aluminium smelter.

An integrated biological effects study using field transplanted mussels was applied to determine the potential biological effects of an effluent discharge from an aluminium smelter into a Norwegian fjord. Chemical body burden and biological effects were measured in mussels positioned downstream (1, 2, 5, 10 and 20 km) from the aluminium smelters discharge for a period of 6 weeks. A suite of biomarkers, from whole organism to subcellular responses were measured. Chemical concentrations in mussel tissues were low; however, a change in the PAC (polyaromatic compound) profile from high to low pyrogenic influence provided evidence of exposure to the smelter's effluent. Overall, the biological responses observed where greater in the mussels positioned closest to the smelter (1-5 km). Lowest chemical accumulation and biomarker responses were observed in mussels positioned 10 km from the smelter and were considered as the reference field population. Mussels located furthest from the smelter (20 km) exhibited significant biomarker responses and suggested a different contaminant source within the fjord. The integrated biological response index (IBR) was applied and reflected the expected level of exposure to the smelters discharge, with highest IBR calculated in mussels positioned closest to the discharge (1-5 km). Principal component analysis (PCA) also differentiated among mussel groups, with the most impacted located closest to the smelter. Not one chemical factor could explain the biological responses observed in mussels, but the presence of PAH16, PAH41 and metals Mn, Ni and Cr were the main contributors measured to the higher stress seen in the mussels from the 1 and 5 km groups.

Characterization of a novel recombinant calcium-binding protein from Arca subcrenata and its anti-hepatoma activities in vitro and in vivo.

Previous studies demonstrated that ASP-3 was a novel calcium-binding protein from Arca subcrenata that effectively inhibited the proliferation of HepG2 cells. To further study the antitumor activity and mechanism of ASP-3, the cytotoxic effects of recombinant ASP-3 were evaluated in HepG2 cells. The results demonstrated that ASP-3 inhibited the proliferation of HepG2 cells by competitively binding to the EGF binding pocket of EGFR and inhibiting the JAK-STAT, RAS-RAF-MEK-ERK, and PI3K-Akt-mTOR signaling pathways mediated by EGFR. ASP-3 significantly inhibited tumor growth in a HepG2 cell subcutaneous xenograft nude mouse model, and its (25 mg/kg and 75 mg/kg) tumor inhibition rates were 46.92 % and 60.28 %, respectively. Furthermore, the crystal structure of ASP-3 was resolved at 1.4 Å. ASP-3 formed as a stable dimer and folded as an EF-Hand structure. ASP-3 stably bound to domain I and domain III of the EGFR extracellular region by using molecular docking and molecular dynamics simulation analysis. Compared with the endogenous ligand EGF, ASP-3 displayed a stronger interaction with EGFR. These experimental results indicated that recombinant ASP-3 possessed an effective anti-hepatoma effect. So, it might be a potential molecule for liver cancer therapy.

Assessment of organic contamination along the coast of Laizhou Bay, China: chemical analysis and integrated biomarker responses in the clam Ruditapes philippinarum.

An investigative biomonitoring study was conducted along the coastal area of Laizhou Bay (China) to evaluate the impact of organic pollution on the clam Ruditapes philippinarum using bioaccumulation and multi-biomarker measurements. In addition, the polychlorinated biphenyls (PCBs), total petroleum hydrocarbons (TPHs) and nonylphenol (NP) content in surface sediment at the study sites were also analyzed. Concentrations of PCBs, TPHs and NP in the sediments of the study area were 1.90 ± 0.10 µg kg-1, 39.55 ± 2.42 mg kg-1, 9.23 ± 0.41 µg kg-1 dry weight, respectively, while the organic contaminants in the soft tissues of R. philippinarum were 14.81 ± 0.96 µg kg-1 for PCBs, 165.87 ± 5.03 mg kg-1 for TPHs and 86.16 ± 5.29 µg kg-1 for NP. Linear regression analysis on the levels of organic pollutants accumulated in R. philippinarum and in sediments showed no significant correlation. Multi-biomarkers including superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase, total glutathione and lipid peroxidation were assayed in gills and digestive glands of R. philippinarum. Finally, the biomarkers in gills were selected to calculate the Integrated Biomarker Response (IBR) index and to evaluate the impact of the three organic contaminants on R. philippinarum collected from different sites. According to IBR results, the western coast and eastern coast exhibited higher environmental stress than the sampling sites along the southern coast of Laizhou Bay. Significant correlation was found between the level of organic contaminants in the sediments and IBR whereas no dependence was found between pollutants' concentrations in sediments and separate biomarker responses. The results showed that PCBs and NP were the main organic pollutants among the three studied which have caused pollution pressure on R. philippinarum in Laizhou Bay coastal area.

Highly-selective complex matrices removal via a modified QuEChERS for determination of triazine herbicide residues and risk assessment in bivalves.

A modified quick, easy, cheap, effective, rugged, and safe (QuEChERs) method for determining triazine herbicide residues in bivalves (Mussels, Scallops, Cockles) was developed. The use of molecularly imprinted polymers (MIPs) as a selective purification material during dispersive-solid phase extraction (d-SPE) increased the removal rate of pigments interference. With 4% acidic acetonitrile as the organic modifier, the modified QuEChERs method achieved good extraction rate of herbicide residues. The satisfactory recoveries (80%-118%) and RSDs (1.0%-11.6%) of herbicide residues were obtained at three spiked levels. The limits of quantification of herbicide residues ranged from 0.10 µg/kg to 1.59 µg/kg. Further, the herbicide residues in bivalves collected in the eastern coasts of China was analyzed. The developed QuEChERs procedure coupled with GC-MS/MS was successfully applied to the herbicide residues detection in bivalves, and due to the extensive use of herbicides and the large consumption of bivalves in globally, the ongoing risk evaluation is needed.

Mussel Shell-Derived Macroporous 3D Scaffold: Characterization and Optimization Study of a Bioceramic from the Circular Economy.

Fish industry by-products constitute an interesting platform for the extraction and recovery of valuable compounds in a circular economy approach. Among them, mussel shells could provide a calcium-rich source for the synthesis of hydroxyapatite (HA) bioceramics. In this work, HA nanoparticles have been successfully synthesized starting from mussel shells (Mytilus edulis) with a two steps process based on thermal treatment to convert CaCO3 in CaO and subsequent wet precipitation with a phosphorus source. Several parameters were studied, such as the temperature and gaseous atmosphere of the thermal treatment as well as the use of two different phosphorus-containing reagents in the wet precipitation. Data have revealed that the characteristics of the powders can be tailored, changing the conditions of the process. In particular, the use of (NH4)2HPO4 as the phosphorus source led to HA nanoparticles with a high crystallinity degree, while smaller nanoparticles with a higher surface area were obtained when H3PO4 was employed. Further, a selected HA sample was synthesized at the pilot scale; then, it was employed to fabricate porous 3D scaffolds using the direct foaming method. A highly porous scaffold with open and interconnected porosity associated with good mechanical properties (i.e., porosity in the range 87-89%, pore size in the range 50-300 µm, and a compressive strength σ = 0.51 ± 0.14 MPa) suitable for bone replacement was achieved. These results suggest that mussel shell by-products are effectively usable for the development of compounds of high added value in the biomedical field.

Occurrence and potential health risks assessment of polycyclic aromatic hydrocarbons (PAHs) in different tissues of bivalves from Hainan Island, China.

The levels of 16 PAHs were determined in the adductor, gills, gonads, hepatopancreas and mantles of the pearl oyster (Pinctada martensii) and the mussel (Perna viridis) collected from coasts of Li'an and Xincun Bays. The levels of ΣPAHs ranged from 597.1 to 2332 ng g-1 d w in the various tissues of bivalves. The pyrolytic source played an important role in the local coastal environment. Significantly higher levels of M-PAHs and H-PAHs were observed in Pinctada martensii than in Perna viridis. The ΣPAHs at different tissues showed the following order from high to low: mantles > hepatopancreas > gonads > gills > adductor. When levels of individual PAHs in the five bivalve tissues have been compared with each other, high correlations have been found (r2 = 0.793-0.975). A general trend was observed that log transformed BSAFs declined with increase of Kow values. The estimated amount of ΣPAHs via ingestion of oyster and mussel varied from 1.35 × 10-2-1.70 × 10-1 and 2.15 × 10-2-1.91 × 10-1 µg kg-1 body weight day-1, respectively. The THQs and CRs calculated for regular consumption of raw bivalves were in the acceptable ranges and may not pose health risk concerns. But for certain population with higher consumption rate for PAHs contaminated bivalves, cautions should be taken for their higher cancer risk.

Assessment of the biochemical and nutritional values of Venerupis decussata from Tunisian lagoons submitted to different anthropogenic ranks.

The present study aimed to evaluate the biochemical status of Venerupis decussata collected seasonally from two Tunisian coastline lagoons (Ghar El Melh and Boughrara) submitted to different anthropogenic levels. Trace elements were significantly higher in clams from Boughrara, especially during the warm season. Fatty acids, proximate compositions, and nutritional value of both clam populations were higher, appearing to be the best for human consumption principally during the spring season. The principal component analysis was determined, including phospholipids and reserve lipids, revealing a great variation with the exogenous and endogenous factors. Generally, clams from Boughrara had a significant alteration in biochemical composition principally due to the anthropogenic levels and physicochemical fluctuation of this lagoon. Our results reflected the healthy benefit of clam consumption and the use of energy reserves and essential fatty acids as a great biomarker to assess ecological risk in lagoons and other coastal ecosystems.

Metal Concentrations in Age-Groups of the Clam, Megapitaria squalida, from a Coastal Lagoon in Mexico: A Human Health Risk Assessment.

The present study shows the human health risk of Cd, Cu, Hg and Zn by consumption of clams Megapitaria squalida from Northwest Mexico, collected in 2013. The mean concentration for each metal in the soft tissue was: Zn > Cu > Cd > Hg; and mean values of 68.89 ± 37.59-30.36 ± 27.19, 8.77 ± 1.35-6.80 ± 0.36, 4.47 ± 0.21-3.18 ± 0.63 and 0.99 ± 0.81-0.52 ± 0.16 µg/g, respectively. Clam age was significantly negatively correlated (p < 0.05) with soft tissue Zn concentrations. For all metals there is a low level of human health risk associated with the consumption of M. squalida, but it is necessary to determine the specific characteristics of the human population of the study site.

Baseline assessment of contaminants in marine biota prior to remediation of industrial effluent impacted sediments in a former tidal estuary in Nova Scotia, Canada.

Contaminated sediments at a pulp mill and former chor-alkali effluent treatment facility in Nova Scotia, Canada will undergo remediation. However, baseline studies assessing contaminants in marine biota in the marine receiving environment are lacking. Historical qualitative and quantitative contaminant data in biota from Boat Harbour (a former tidal lagoon which was used to treat industrial effluent since 1967), and surrounding marine environment were reviewed to establish baseline pollution from industrial effluent and contaminated sediments. Elevated metal, dioxins and furan concentrations previously measured in marine biota needs updating to help inform pre-remediation monitoring. Selection of species, contaminants of concern and sampling locations were ad hoc and often inconsistent with environmental effects monitoring requirements under Canadian federal Pulp and Paper Effluent Regulations. These consolidated baseline data are required to determine historical impacts and to assist future monitoring during Boat Harbour sediment remediation to compare against.

Psychoactive substances in mussels: Analysis and occurrence assessment.

This work presents an analytical methodology based on a "Quick, easy, cheap, effective, rugged, and safe" (QuEChERS) extraction and liquid chromatography-tandem mass spectrometry detection (LC-MS/MS) for the simultaneous determination of 35 psychoactive substances in fresh mussel samples. The compounds investigated include illicit drugs, new psychoactive substances, commonly prescribed psychiatric pharmaceuticals, and caffeine. The methodology was validated in terms of recovery (relative recoveries 77-118%), repeatability (relative standard deviation values <20%), and sensitivity (limits of detection and quantification <2 ng/g fresh weight (f.w.) and <6.7 ng/g f.w., respectively, for most compounds). The method was applied to the analysis of 15 samples, covering both commercially available mussels purchased from local food markets and wild fresh mussels collected in the Northeast coast of Spain in the Mediterranean Sea. Only one sample corresponding to wild mussels was found to contain 2 of the target analytes, namely, sertraline (1.5 ng/g f.w.) and caffeine, (12.8 ng/g f.w.).

Assessment of trace metal contamination in the marine sediment, seawater, and bivalves of Parangipettai, southeast coast of India.

Industrial and domestic discharge into the coastal environment has driven us to propose an integrated approach to delineate stations contaminated with metals on the Parangipettai coast by collecting sediment and seawater samples monthly at 18 stations from 2015 to 2017. Descriptive statistics revealed that the concentrations of some metals in the sediment and seawater samples were beyond the permissible level. Further, factor analysis showed a sampling adequacy of 0.90 with high positive loading for Ni (0.94), Cd (0.91), Co (0.90), Pb (0.89), and Zn (0.87) in sediment samples. The degree of contamination by metals was evaluated using pollution indices. The results of the contamination index revealed that some stations in the study area were moderately polluted, and those of the ecological index showed that open sea was under low risk while other stations were in the moderate-to-high-risk category. The results obtained are essential to establish the reference condition for a comparative study in similar environments in the tropical regions.

Market Basket Survey of Arsenic Species in the Top Ten Most Consumed Seafoods in the United States.

The study focused on the determination of arsenic species in the top ten most consumed seafoods in the United States. Fifty-four samples were collected from local supermarkets, and their species identities were confirmed by DNA barcoding. The total arsenic in the samples varied greatly in the range of 8-22200 ng/g (wet mass). Speciation analysis based on extraction of water-soluble and nonpolar arsenic showed that inorganic arsenic (iAs) was found only in clams and crabs, while arsenobetaine (AsB) predominates in most samples. Among the other arsenicals, trimethylarsoniopropionate (TMAP) was found in most matrices with higher concentrations in crabs, and arsenosugars existed in most clams and crabs. Nonpolar arsenic accounted for 1-46% of the total arsenic in the samples. The accuracy of the analytical results was evaluated using standard reference materials and spike recovery tests. The survey showed that the iAs concentrations in America's most consumed seafood products are much lower than the tolerable intake set by the Joint FAO/WHO Expert Committee, even at the highest levels found in this study.

Mercury and Po-210 in mollusc species in the island of Gökçeada in the north-eastern Aegean Sea: Bioaccumulation and risk assessment for human consumers.

Among the radioactive pollutants 210Po is the most substantial one in terms of seafood safety due to its efficient accumulation in marine animals and high irradiation of its alpha emission. Mercury is a highly toxic metal for both marine organisms and human beings. Biomagnification of MeHg (methylmercury) through marine food chains has made Hg concern of ecotoxicology and seafood safety. In the current study, the bioaccumulation of 210Po and THg (total mercury) were determined in 20 mollusc species, including 8 bivalves, 7 gastropods and 5 cephalopods collected from the island of Gökçeada in the north-eastern Aegean Sea. The highest accumulation of 210Po and Hg was seen in bivalves and cephalopods, respectively. Elevated Hg concentrations in all body parts (arms, mantle and viscera) were observed in octopus' species. The results of this study suggests that filter feeder bivalves and gastropods have a capacity to concentrate 210Po in their bodies, whereas predator gastropods and cephalopods have a capacity to concentrate Hg in their bodies. 7.0 kg (3.2-14.2) bivalve flesh intake is adequate due to 210Po ingestion in the studied region to reach 1 mSv which is the annual committed effective dose. Octopus consumption of 705 g in a week alone is needed to reach Provisional Tolerable Weekly Intake (PTWI) of mercury, 5 µg kg-1 body weight. Due to very low non-fish seafood consumption in Turkey there is no risk of Hg intake and alpha radiation of 210Po above the limit values through mollusc consumption.

Dietary Intake of Essential, Toxic, and Potentially Toxic Elements from Mussels (Mytilus spp.) in the Spanish Population: A Nutritional Assessment.

The levels of forty-three elements were determined in fresh, preserved, and frozen mussels (n = 208) with the purpose of evaluating their contribution to the recommended dietary intake of essential elements and their potential risk to Spanish consumers' health. We found relevant differences in the element content in relation to the mode of conservation of mussels as well as in relation to their geographical origin, brand, or mode of production. According to our estimates, mussels are important contributors to the intake of most essential elements, contributing almost 70% of daily requirements of Se, 30-35% of Mo, Zn, and Co, and around 15% of Fe. At the same time, the pattern of average consumption of mussels in Spain does not seem to imply an excessive risk associated with any of the 36 toxic elements studied. However, it should be noted that, in the high percentile of consumption the exposure to Cd and As may be high, in particular that associated with the consumption of fresh and/or frozen mussels. According to the results of this study, a moderate consumption of mussels can be recommended as a valuable and safe source of trace elements.

Linking social preferences and ocean acidification impacts in mussel aquaculture.

Ocean Acidification (OA) has become one of the most studied global stressors in marine science during the last fifteen years. Despite the variety of studies on the biological effects of OA with marine commercial species, estimations of these impacts over consumers' preferences have not been studied in detail, compromising our ability to undertake an assessment of market and economic impacts resulting from OA at local scales. Here, we use a novel and interdisciplinary approach to fill this gap. We experimentally test the impact of OA on commercially relevant physical and nutritional attributes of mussels, and then we use economic discrete choice models to assess the marginal effects of these impacts over consumers' preferences and wellbeing. Results showed that attributes, which were significantly affected by OA, are also those preferred by consumers. Consumers are willing to pay on average 52% less for mussels with evidences of OA and are willing to increase the price they pay to avoid negative changes in attributes due to OA. The interdisciplinary approach developed here, complements research conducted on OA by effectively informing how OA economic impacts can be analyzed under the lens of marginal changes in market price and consumer' welfare. Thereby, linking global phenomena to consumers' wellbeing, and shifting the focus of OA impacts to assess the effects of local vulnerabilities in a wider context of people and businesses.

Cost-Effective and High-Reliability Analytical Approach for Multitoxin Screening in Bivalve Mollusks by Liquid Chromatography Coupled to Tandem Mass Spectrometry.

A fast, less expensive, analytical approach with high metrologic reliability was developed to assist an official program for 21 marine biotoxins, monitoring in bivalve mollusks. The simultaneous analysis of lipophilic and hydrophilic marine biotoxins was achieved using a sample preparation protocol based on solid-liquid extraction and low-temperature cleanup, followed by liquid chromatography coupled to tandem mass spectrometry. Samples were extracted with acidified methanol/water (90:10), followed by low-temperature cleanup. Chromatographic separation was obtained using a cyano-bonded silica phase. The mobile phase was composed of water and acetonitrile, with both 0.1% formic acid and 2.5 mmol L-1 ammonium formate. Electrospray ionization was used in both negative and positive modes. The single-laboratory validation approach enabled method performance assessment, and the necessary data to design a model for result expression were yielded. With this purpose, a systematic study of errors and uncertainties was performed. This new analytical approach aimed to minimize the use of highly expensive analytical standards, promoting economic viability to be applied by high-throughput routine laboratories. After its implementation on the Brazilian official monitoring program, positive results near the regulatory limits were obtained, demonstrating the fit for purpose of the method as a surveillance tool.