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Pharmaceuticals are recognised as environmental contaminants of emerging concern (CECs) due to their increasing presence in the aquatic environment, along with high bioactivity linked to their therapeutic use. Therefore, information on environmental levels is urgently required. This study examined the presence of a range of common pharmaceuticals in oysters and mussels intended for human consumption from England and Wales using stable isotope dilution tandem mass spectrometry. A range of compounds were detected in bivalve tissue, with the Selective Serotonin Reuptake Inhibitor antidepressant sertraline being most abundant, reaching a maximum concentration of 22.1 ng/g wet weight shellfish tissue. Levels of all pharmaceuticals showed seasonal and geographical patterns. A dietary risk assessment revealed that the levels of pharmaceuticals identified in bivalve molluscs represent a clear hazard, but not a risk for the consumer. This study highlights the requirement for further monitoring of the presence of pharmaceuticals and other CECs in bivalve molluscs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Crown Copyright © 2024. Published by Elsevier Ltd. All rights reserved.)

Santa Catarina is the main producer state of oysters and mussels in Brazil, reaching 98 % of national production. To assure the safety of bivalve mollusks production, control programs of marine biotoxins (MBs) have been continuously performed. Herein, the co-occurrence of MBs and contaminants of emerging concern (CECs) in oyster and mussels from the main production sites of Santa Catarina was reported, covering 178 compounds. Samples of wild and non-cultivated oysters and mussels were also assessed. Chemometric tools were used to evaluate and optimize several sample preparation techniques such as solid-liquid, ultrasound assisted, and pressurized liquid extraction. The optimized protocol was based on ultrasound assisted extraction followed by liquid chromatography coupled to tandem mass spectrometry. The results showed the incidence of several CECs and MBs. In the case of MBs, all results were below the regulatory limits for both cultivated and non-cultivated samples. Wild mollusks have shown a higher number of compounds. Regarding CECs, the more frequent compounds were caffeine, diclofenac, meloxicam, and sertraline. Domoic acid and okadaic acid were the main toxins detected. The results highlighted the need of monitoring for MBs and the potential of oyster and mussels as sentinel organisms to risk analysis of CECs in coastal regions. To the best of our knowledge, this is the first method to describe a simultaneous sample preparation and analysis of CECs and MBs in bivalve mollusks, as well as the first report of meloxicam and florfenicol in mussels and oysters., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Background: Given the recent detection of tetrodotoxin (TTX) in bivalve molluscs but the absence of a full collaborative validation study for TTX determination in a large number of shellfish samples, interlaboratory assessment of method performance was required to better understand current capabilities for accurate and reproducible TTX quantitation using chemical and immunoassay methods., Objective: The aim was to conduct an interlaboratory study with multiple laboratories, using results to assess method performance and acceptability of different TTX testing methods., Methods: Homogenous and stable mussel and oyster materials were assessed by participants using a range of published and in-house detection methods to determine mean TTX concentrations. Data were used to calculate recoveries, repeatability, and reproducibility, together with participant acceptability z-scores., Results: Method performance characteristics were good, showing excellent sensitivity, recovery, and repeatability. Acceptable reproducibility was evidenced by HorRat values for all LC-MS/MS and ELISA methods being less than the 2.0 limit of acceptability. Method differences between the LC-MS/MS participants did not result in statistically different results. Method performance characteristics compared well with previously published single-laboratory validated methods and no statistical difference was found in results returned by ELISA in comparison with LC-MS/MS., Conclusion: The results from this study demonstrate that current LC-MS/MS methods and ELISA are on the whole capable of sensitive, accurate, and reproducible TTX quantitation in shellfish. Further work is recommended to expand the number of laboratories testing ELISA and to standardize an LC-MS/MS protocol to further improve interlaboratory precision., Highlights: Multiple mass spectrometric methods and a commercial ELISA have been successfully assessed through an interlaboratory study, demonstrating excellent performance., (© The Author(s) 2023. Published by Oxford University Press on behalf of AOAC INTERNATIONAL.)

A single laboratory method performance verification is reported for a rapid sensitive UHPLC-MS/MS method for the quantification of eight cyclic imine and two brevetoxin analogues in two bivalve shellfish matrices: mussel (Mytilus edulis) and Pacific oyster (Crassostrea gigas). Targeted cyclic imine analogues were from the spirolide, gymnodimine and pinnatoxin groups, namely 20-Me-SPX-C, 13-desMe-SPX-C, 13,19-didesMe-SPX-C, GYM-A, 12-Me-GYM, PnTx-E, PnTx-F and PnTx-G. Brevetoxin analogues consisted of the shellfish metabolites BTX-B5 and S-desoxy-BTX-B2. A rapid dispersive extraction was used as well as a fast six-minute UHPLC-MS/MS analysis. Mobile phase prepared using ammonium fluoride and methanol was optimised for both chromatographic separation and MS/MS response to suit all analytes. Method performance verification checks for both matrices were carried out. Matrix influence was acceptable for the majority of analogues with the MS response for all analogues being linear across an appropriate range of concentrations. In terms of limits of detection and quantitation the method was shown to be highly sensitive when compared with other methods. Acceptable recoveries were found with most analogues, with laboratory precision in terms of intra- and inter-batch precision deemed appropriate. The method was applied to environmental shellfish samples with results showing low concentrations of cyclic imines to be present. The method is fast and highly sensitive for the detection and quantification of all targeted analogues, in both mussel and oyster matrices. Consequently, the method has been shown to provide a useful tool for simultaneous monitoring for the presence or future emergence of these two toxin groups in shellfish., (Crown Copyright © 2021. Published by Elsevier B.V. All rights reserved.)

With the extensive application of nanotechnology, metal nanoparticles (MNPs) have been widely used, thus are universally detected in the environment. This has caused increasingly concerns due to their toxicity and the potential health risks they pose to humans. In this work, the concentrations and particle size distributions of MNPs and concentrations of associated metal ionic species in shellfish seafood (clams and oysters) were investigated using single particle inductively coupled plasma mass spectrometry (sp-ICP-MS) and inductively coupled plasma mass spectrometry (ICP-MS). The MNPs in the clam and oyster tissues were extracted via an alkaline digestion method with a recovery rate of 95.9% (for gold nanoparticles (AuNPs)). Then total concentrations of 41 metal elements were measured in the two types of seafood, of which 20 were selected for sp-ICP-MS analysis. The results showed that 5 types of MNPs were detectable in clams (Y, La, Ce, Pr, Gd) and 5 types of MNPs were detectable in oysters (Y, La, Ce, Pr, Nd). Size distributions of MNPs in clams and oysters were in the range of 35-55 nm and 30-65 nm, respectively. Nanoparticle concentrations in clams and oysters ranged from 0.6 to 37.7 ng/g and 4.2-19.7 ng/g, and accounted for 3.4%-50% and 5.5%-46% of the total metal content, respectively. Based on this analysis, the health risks of metals in the two kinds of seafood were evaluated by comparing the Provisional Tolerable Weekly Intake (PTWI) with limits recommended by the World Health Organization (WHO)/Food and Agriculture Organization (FAO). These results provide important information about the presence of metal nanoparticles in seafood and, to the best of our knowledge, this is the first time that the nanoparticles of rare earth elements have been detected and reported in bivalve mollusc tissues., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Inc. All rights reserved.)

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 (r 2  = 0.793-0.975). A general trend was observed that log transformed BSAFs declined with increase of K ow 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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The profile of 11 trace metals in two commonly used biomonitors (the native oyster Crassostrea palmula and mussel Mytella strigata) from Estero de Urias lagoon, Gulf of California, were studied for six months, covering both dry and wet seasons. Metal concentrations in these two bivalves were compared with concentrations accumulated by Artificial Mussels (AMs) deployed alongside during the same period. Significant temporal variations in Cd, Cr and Mn were observed in both bivalve species and AMs. Temporal changes were observed for Fe in both bivalve species, Pb in oyster only and Cu in both AMs and oysters, revealing seasonal changes in inputs and/or chemical forms of these metals in the lagoon. Significant correlations for Cd, Cr and Cu were found in mussels and oysters, but their Co, Fe, Mn and Zn profiles were very different, despite these two species being taxonomically closely related and often used as biomonitors for metals. Interestingly, Hg and U were detected in AMs but not in oysters and mussels. The difference in metal profile in oysters, mussels and AMs revealed in the present study clearly showed that different biomonitors and AM take up metals differentially from the same environment, and metal profile in a single biomonitor or AM alone therefore, cannot provide a good estimate on metal concentrations in the ambient environment. As such, different biomonitors and AM should be used in metal monitoring, in order to provide a comprehensive picture on metal levels in aquatic ecosystems. Concentrations of Ni and Pb in oysters, and Cr, Fe and Mn in mussels were among the highest reported in coastal waters worldwide. Concentrations of Pb in oysters exceeded legal limits set for bivalve mollusks in EU. Concentrations of Cr in mussels and oysters exceeded or were very close to, respectively, the legal limit for fish, crab-meat, oysters, prawns, and shrimps in Hong Kong. The results indicate a potential public health risk on human consumption of oysters and mussels commonly harvested from the Estero de Urias lagoon, and corresponding pollution control measures are deemed necessary., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

When considering the geographical expansion of marine toxins, the emergence of new toxins and the associated risk for human health, there is urgent need for versatile and efficient analytical methods that are able to detect a range, as wide as possible, of known or emerging toxins. Current detection methods for marine toxins rely on a priori defined target lists of toxins and are generally inappropriate for the detection and identification of emerging compounds. The authors describe the implementation of a recent approach for the non-targeted analysis of marine toxins in shellfish with a focus on a comprehensive workflow for the acquisition and treatment of the data generated after liquid chromatography coupled with high resolution mass spectrometry (LC-HRMS) analysis. First, the study was carried out in targeted mode to assess the performance of the method for known toxins with an extended range of polarities, including lipophilic toxins (okadaic acid, dinophysistoxins, azaspiracids, pectenotoxins, yessotoxins, cyclic imines, brevetoxins) and domoic acid. The targeted method, assessed for 14 toxins, shows good performance both in mussel and oyster extracts. The non-target potential of the method was then challenged via suspects and without a priori screening by blind analyzing mussel and oyster samples spiked with marine toxins. The data processing was optimized and successfully identified the toxins that were spiked in the blind samples.

The objective of this study was to determine the effect of high hydrostatic pressure (HHP) on the inactivation of Vibrio sp. in pure cultures and mussel homogenates. Four Vibrio strains including V. alginolyticus, V. cholerae, V. parahaemolyticus and V. vulnificus were used. HHP treatments were performed with both pure Vibrio sp. cultures in alkaline peptone water (2% NaCl) and artificially inoculated mussel homogenates at pressure levels of 250, 350 and 450 MPa for 1 and 3 min at 25°C. Counts of Vibrio were determined before and after treatment using drop plating method. The effect of high pressure on the reduction level significantly differed among the respective Vibrio species. Vibrio vulnificus was the most susceptible species to HHP. To achieve a >5 log reduction in mussel homogenates, pressure treatment needs to be (i) 350-450 MPa for ≥1 min at 25°C for both V. alginolyticus and V. cholerae, (ii) 250 MPa for ≥3 min or 350-450 MPa for ≥1 min for V. vulnificus and (iii) 350 MPa for ≥3 min or 450 MPa for ≥1 min for V. parahaemolyticus., Significance and Impact of the Study: High hydrostatic pressure (HHP) has been applied to inactivate spoilage and pathogenic micro-organisms in a variety of food products, including seafood. Vibrio sp. are frequently reported as the main cause of foodborne illness associated with consumption of raw or undercooked seafood particularly shellfish worldwide. To date, data on the inactivation of Vibrio sp. via HHP are still limited and most of the trials only investigated HHP application in oysters and clams. This study demonstrates the efficacy of HHP inactivating Vibrio sp. in both pure culture and mussel homogenates., (© 2018 The Society for Applied Microbiology.)

Shellfish is a common part of indigenous cuisines throughout the world and one of the major sources of human exposure to arsenic (As). We evaluated As speciation in shellfish after cooking and gastrointestinal digestion in this study. Results showed that washing and cooking (boiling and steaming) can reduce As exposures from shellfish. The use of spices during cooking processes also helped to reduce the bioaccessibility of total As. Through mass balance calculations, we verified the transformation of methylated As compounds into inorganic As in shellfish takes place during cooking and that As demethylation can occur during simulated gastrointestinal digestion. In vivo demethylation of As after gastrointestinal digestion was also demonstrated in laboratory mice. This increase in inorganic As during digestion suggests that risks of As toxicity from shellfish consumption are being underestimated. Further studies on the mechanisms of As speciation transformation in food are necessary for more thorough risk assessments.

A receptor binding assay (RBA) for the determination of paralytic shellfish poisoning toxicity is formally validated through collaborative study and approved for regulatory monitoring use in the US for mussels and clams. However, to date, the method has not been tested on bivalve molluscs originating from European waters and no validation studies have been conducted for oysters, a shellfish species of great importance globally. This study firstly reports the work conducted to assess the performance of the assay in comparison with a regulatory chemical detection method for a range of shellfish species originating from Great Britain. Data obtained showed a complete absence of false negative RBA results, with a tendency to over-estimate PSP toxicity for some shellfish species in comparison with liquid chromatography with fluorescence detection. Secondly, the performance of the RBA was assessed for oysters, with the analysis of a dilution series of oyster matrix certified reference materials. Method trueness, sensitivity and precision were found to compare well with results reported previously for other species. In addition, the RBA analysis of untreated and demetallated oyster extracts, provided good evidence that the RBA is not suppressed in the presence of high concentrations of zinc as reported previously for the mouse bioassay. Consequently, there is strong evidence from this study, that the RBA would be suitable for determination of PSP toxicity in bivalve molluscs from GB, with acceptable method performance in oysters. Further validation studies would be required for other shellfish species of interest before the method can be considered suitable for implementation in Europe., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Aquaculture, which is growing 3-5 times faster than terrestrial agriculture, will play an important role to meet future global food production needs. However, over 80% of global sewage production is returned to the environment untreated or poorly treated. In developing nations, these nontraditional waters of diverse quality are being recycled for aquaculture, yet chemical residues are differentially studied. Here, we examined pharmaceuticals, pesticides, polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), polycyclic aromatic hydrocarbons (PAHs), and flame retardants in marine bivalves using isotope dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS) and low-pressure gas chromatography-tandem mass spectrometry (LP GC-MS/MS). Green-lipped mussels from the field and oysters from aquaculture net pens, which are harvested as food products, were collected adjacent to point source municipal wastewater and landfill leachate effluent discharges, respectively, in Hong Kong, the fourth most densely populated country in the world. Multiple classes of pharmaceutical, pesticides, PAHs, and phosphorus-based flame retardants were detected at low μg/kg levels. Acceptable servings per week indicated minimal risk for a number of chemicals; however, such calculations could not be performed for other contaminants of emerging concern. Future efforts are needed to better understand contaminant influences on marine bivalve populations and aquaculture product safety, particularly in rapidly urbanizing regions of developing countries with limited wastewater infrastructure.

The occurrence of freshwater harmful algal bloom toxins impacting the coastal ocean is an emerging threat, and the potential for invertebrate prey items to concentrate toxin and cause harm to human and wildlife consumers is not yet fully recognized. We examined toxin uptake and release in marine mussels for both particulate and dissolved phases of the hepatotoxin microcystin, produced by the freshwater cyanobacterial genus Microcystis. We also extended our experimental investigation of particulate toxin to include oysters (Crassostrea sp.) grown commercially for aquaculture. California mussels (Mytilus californianus) and oysters were exposed to Microcystis and microcystin toxin for 24h at varying concentrations, and then were placed in constantly flowing seawater and sampled through time simulating riverine flushing events to the coastal ocean. Mussels exposed to particulate microcystin purged the toxin slowly, with toxin detectable for at least 8 weeks post-exposure and maximum toxin of 39.11ng/g after exposure to 26.65μg/L microcystins. Dissolved toxin was also taken up by California mussels, with maximum concentrations of 20.74ng/g after exposure to 7.74μg/L microcystin, but was purged more rapidly. Oysters also took up particulate toxin but purged it more quickly than mussels. Additionally, naturally occurring marine mussels collected from San Francisco Bay tested positive for high levels of microcystin toxin. These results suggest that ephemeral discharge of Microcystis or microcystin to estuaries and the coastal ocean accumulate in higher trophic levels for weeks to months following exposure., (Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.)

The analytical procedures for reference measurements of the total Hg and methyl mercury (MeHg) mass fractions at various concentration levels in marine biota samples, candidates for certified reference materials (oyster and clam Gafrarium tumidum), were evaluated. Two modes of application of isotope dilution inductively coupled plasma mass spectrometry method (ID ICP-MS), namely direct isotope dilution and species-specific isotope dilution analysis with the use of two different quantification mass spectrometry techniques were compared. The entire ID ICP-MS measurement procedure was described by mathematical modelling and the combined uncertainty of measurement results was estimated. All factors influencing the final results as well as isotopic equilibrium were systematically investigated. This included the procedural blank, the moisture content in the biota samples and all factors affecting the blend ratio measurements (instrumental background, spectral interferences, dead time and mass discrimination effects as well as the repeatability of measured isotopic ratios). Modelling of the entire measurement procedures and the use of appropriate certified reference materials enable to assure the traceability of obtained values to the International System of Units (SI): the mole or the kilogram. The total mass fraction of mercury in oyster and clam biota samples, after correction for moisture contents, was found to be: 21.1 (1.1) 10(-9) kg kg(-1) (U =5.1% relative, k=2) and 390.0 (9.4) 10(-9) kg kg(-1) (U=2.4% relative, k=2), respectively. For the determination of mercury being present as methyl mercury, the non-chromatographic separation on anion-exchange resin AG1-X8 of the blended samples was applied. The content of MeHg (as Hg) in oyster sample was found: 4.81 (24) 10(-9)kgkg(-1) (U=5.0%, k=2) and 4.84 (21) 10(-9)kgkg(-1) (U=4.3%, k=2) with the use of quadrupole (ICP QMS) or sector field (ICP SFMS) inductively coupled plasma mass spectrometers, respectively. In the case of clam sample, the concentration of MeHg (as Hg) was found to be: 61.0 (2.3) 10(-)(9)kgkg(-1) (U=3.8%, k=2) and 61.3 (2.2) 10(-)(9)kgkg(-1) (U=3.6%, k=2), respectively. The mass fractions for total Hg and MeHg determined in this study were used as a contribution of the International Atomic Energy Agency (IAEA) Environment Laboratories in the characterisation of the IAEA 461 and IAEA 470 certified reference materials. The obtained good agreement with the reference values further validated the methods developed in this study., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The concentrations of As, Cd, Cu, Hg, Pb and Zn were measured in four shellfish species (oysters, mussels, scallops, warty venus) collected along the Istria Peninsula coast during autumn 2013. Mean metal levels in shellfish ranged as follows (mg/kg): As 1.69-3.91, Cd 0.15-0.99, Hg 0.06-0.2, Pb 0.33-0.74, Cu 1.23-36.2, Zn 16.3-511. Statistical analysis showed a significant difference in As (p<0.05), Cd, Pb, Hg, Cu and Zn (p<0.01, all) levels among shellfish species. The highest metal concentrations were found in oysters (mg/kg): As 3.91, Cd 0.99, Cu 36.2 and Zn 511. The highest means of Pb and Hg were measured in (mg/kg): warty venus 0.74; scallops 0.12. The estimated daily intakes for oysters shown it as a good source of Cu and Zn. The higher contribution of Cd and As levels to the provisional tolerable weekly intake in shellfishes suggest a careful approach for consumption.

A single-step lateral flow immunoassay was developed and validated to detect okadaic acid (OA) and dinophysis toxins (DTXs), which cause diarrhetic shellfish poisoning. The performance characteristics of the test were investigated, in comparison to reference methods (liquid chromatography tandem mass spectrometry and/or bioassay), using both spiked and naturally contaminated shellfish. A portable reader was used to generate a qualitative result, indicating the absence or presence of OA-group toxins, at concentrations relevant to the maximum permitted level (MPL). Sample homogenates could be screened in 20 min (including extraction and assay time) for the presence of free toxins (OA, DTX1, DTX2). DTX3 detection could be included with the addition of a hydrolysis procedure. No matrix effects were observed from the species evaluated (mussels, scallops, oysters, and clams). Results from naturally contaminated samples (n = 72) indicated no false compliant results and no false noncompliant results at <50% MPL. Thus, the development of a new low-cost but highly effective tool for monitoring a range of important phycotoxins has been demonstrated.

A single-step lateral flow immunoassay (LFIA) was developed and validated for the rapid screening of paralytic shellfish toxins (PSTs) from a variety of shellfish species, at concentrations relevant to regulatory limits of 800 μg STX-diHCl equivalents/kg shellfish meat. A simple aqueous extraction protocol was performed within several minutes from sample homogenate. The qualitative result was generated after a 5 min run time using a portable reader which removed subjectivity from data interpretation. The test was designed to generate noncompliant results with samples containing approximately 800 μg of STX-diHCl/kg. The cross-reactivities in relation to STX, expressed as mean ± SD, were as follows: NEO: 128.9% ± 29%; GTX1&4: 5.7% ± 1.5%; GTX2&3: 23.4% ± 10.4%; dcSTX: 55.6% ± 10.9%; dcNEO: 28.0% ± 8.9%; dcGTX2&3: 8.3% ± 2.7%; C1&C2: 3.1% ± 1.2%; GTX5: 23.3% ± 14.4% (n = 5 LFIA lots). There were no indications of matrix effects from the different samples evaluated (mussels, scallops, oysters, clams, cockles) nor interference from other shellfish toxins (domoic acid, okadaic acid group). Naturally contaminated sample evaluations showed no false negative results were generated from a variety of different samples and profiles (n = 23), in comparison to reference methods (MBA method 959.08, LC-FD method 2005.06). External laboratory evaluations of naturally contaminated samples (n = 39) indicated good correlation with reference methods (MBA, LC-FD). This is the first LFIA which has been shown, through rigorous validation, to have the ability to detect most major PSTs in a reliable manner and will be a huge benefit to both industry and regulators, who need to perform rapid and reliable testing to ensure shellfish are safe to eat.

Acquiring antimicrobial-resistant (AMR) bacteria through consuming contaminated animal food products is an emerging public health concern, though the sources of contamination are not always clear. This study characterized the occurrence of AMR in Escherichia coli from bivalve molluscs and assessed for the possible sources in the Hillsborough river complex of Prince Edward Island, Canada in areas overlapping with an oyster fishery. Multivariable statistical analysis indicated that the probability of detecting E. coli increased as the estimated dosage of animal effluent contamination decreased. Isolates with AMR were only found from sampling sites closest to untreated human effluent sources. Twenty-seven percent (n=6 of 22) of the isolates were pathogenic, with virulence factors consistent with extraintestinal E. coli of human origin. Though there is more evidence of contamination arising from human effluent, more research is needed to identify driving sources.

Estuarine shorelines have been degraded since humans arrived in the coastal zone. In recent history, a major cause of habitat degradation has been the armoring of shorelines with vertical walls to protect property from erosive wave energy; however, a lack of practical alternatives that maintain or enhance ecological function has limited the options of waterfront residents and coastal zone managers. We experimentally investigated the habitat value of two configurations of submerged breakwaters constructed along an eroding shoreline in northwest Mobile Bay, AL (USA). Breakwaters comprised of bagged oyster shell or Reef Ball™ concrete domes were built by a community-based restoration effort. Post-deployment monitoring found that: bagged oyster breakwaters supported much higher densities of live ribbed mussels than Reef Ball breakwaters; both breakwater configurations supported increased species richness of juvenile and smaller fishes compared to controls; and that larger fishes did not appear to be affected by breakwater presence. Our study demonstrates that ecologically degraded shorelines can be augmented with small-scale breakwaters at reasonable cost and that these complex structures can serve as habitat for filter-feeding bivalves, mobile invertebrates, and young fishes. Understanding the degree to which these structures mitigate erosive wave energy and protect uplands will require a longer time frame than our 2-year-long study.

A liquid chromatography quadrupole linear ion trap mass spectrometry method with fast polarity switching and a scheduled multiple reaction monitoring algorithm mode was developed for multiclass screening and identification of lipophilic marine biotoxins in bivalve molluscs. A major advantage of the method is that it can detect members of all six groups of lipophilic marine biotoxins [okadaic acid (OA), yessotoxins (YTX), azaspiracids (AZA), pectenotoxins (PTX), cyclic imines (CI), and brevetoxins (PbTx)], thereby allowing quantification and high confidence identification from a single liquid chromatography tandem mass spectrometry (LC-MS/MS) injection. An enhanced product ion (EPI) library was constructed after triggered collection of data via information-dependent acquisition (IDA) of EPI spectra from standard samples. A separation method for identifying 17 target toxins in a single analysis within 12min was developed and tested. Different solid phase extraction sorbents, the matrix effect (for oyster, scallop, and mussel samples), and stability of the standards also were evaluated. Matrix-matched calibration was used for quantification of the toxins. The limits of detection were 0.12-13.6μg/kg, and the limits of quantification were 0.39-45.4μg/kg. The method was used to analyze 120 shellfish samples collected from farming areas along the coast of China, and 7% of the samples were found to be contaminated with toxins. The library search identified PbTx-3, YTX, OA, PTX2, AZA1, AZA2, and desmethylspirolide C (SPX1). Overall, the method exhibited excellent sensitivity and reproducibility, and it will have broad applications in the monitoring of lipophilic marine biotoxins., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Surface sediment and biota were collected from 12 sampling sites - seven along the Pearl River Delta and five along the Hong Kong coastline. Perfluorinated compound (PFC) concentrations were detected using a high-performance-liquid-chromatogram-tandem-mass-spectrometry system. Analytical results indicated that the total PFC concentrations were in the range of 0.15-3.11 ng/g dry weight in sediments, while the total PFC concentrations in oyster and mussel samples were between 0.46-1.96 and 0.66-3.43 ng/g wet weight, respectively. The major types of PFCs detected in the sediment samples were perfluorooctanesulfonic acid (PFOS) and perfluorobutanoic acid (PFBA), with concentrations ranging from low limits of quantification to 0.86±0.12 ng/g dry weight and 1.50±0.26 ng/g dry weight, respectively. In bivalve samples, PFOS was the dominant contaminant with concentrations ranging from 0.25±0.09 to 0.83±0.12 ng/g wet weight in oysters and 0.41±0.14 to 1.47±0.25 ng/g wet weight in mussels. An increase in PFC concentration was found to be correlated with increased human population density in the study areas., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The bioaccumulation of polycyclic aromatic hydrocarbons (PAHs), organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) was characterized in cockle, oyster and noble pen shell from nine areas in the Mar Menor lagoon with different hydrodynamic and pollutant sources. Biota, sediment and water samples were simultaneously collected in the spring and autumn of 2010. Considering all bivalve samples, PAH concentrations ranged from 8.98 to 370 μg·kg(-1) d.w., those of PCBs from 0.15 to 42.36 μg·kg(-1) d.w. and those of DDXs from below detection limit to 240.6 μg·kg(-1) d.w., where p,p'-DDE was the main fraction. The bioaccumulation of PAHs was similar for cockle, oyster and noble pen shell, being higher close to ports and wastewater effluents. However, DDX and PCB bioaccumulations in oyster and noble pen shell were significantly higher than in cockle in spring (p=0.02). The first organic pollutant bioaccumulation data for noble pen shell were obtained in this study, showing a preferential accumulation of pyrene. The increase of PAH bioaccumulation in autumn, as compared to spring, was low, due to high water temperatures during the summer, which favoured PAH dissipation processes. No significant seasonal variations were detected for OCPs and PCBs, except in some specific areas. The PAH, PCB and OCP levels detected in these bivalves were lower than OSPAR/MED POL environmental assessment criteria, except for p,p'-DDE in bivalves sited close to El Albujón watercourse mouth., (© 2013 Elsevier B.V. All rights reserved.)

Thirteen laboratories participated in an inter-laboratory study to evaluate the method performance characteristics of a liquid chromatography-tandem mass spectrometric method (LC-MS/MS) for marine lipophilic shellfish toxins. Method performance characteristics were evaluated for mussel (Mytilus edulis), oyster (Crassostrea gigas) and cockle (Cerastoderma edule) matrices. The specific toxin analogues tested included okadaic acid (OA), dinophysistoxins-1 and -2 (DTX1, -2), azaspiracids-1, -2 and -3 (AZA1, -2, -3), pectenotoxin-2 (PTX2), yessotoxin (YTX), and 45-OH-yessotoxin (45-OH-YTX). The instrumental technique was developed as an alternative to the still widely applied biological methods (mouse or rat bioassay). Validation was conducted according to the AOAC-harmonised protocol for the design, conduct and interpretation of method-performance studies. Eight different test materials were sent as blind duplicates to the participating laboratories. Twelve laboratories returned results that were accepted to be included in the statistical evaluation. The method precision was expressed as HORRATs. For the individual toxins (except for 45-OH-YTX) HORRATs were found to be ≤1.8 (median HORRAT=0.8) in all tested materials. The recoveries of OA-, AZA- and YTX-group toxins were within the range of 80-108% and PTX2 was within the range of 62-93%. Based on the acceptable values for precision and recovery, it was concluded that the method is suitable for official control purposes to quantitatively determine OA/DTXs, AZAs, YTXs and PTX2 in shellfish., (© 2011 Taylor & Francis)

To better understand critical aspects of diarrhetic shellfish poisoning (DSP) occurrence in a chief producing region of bivalves in Korea, the geographical and annual variation of DSP toxins and other lipophilic toxins in mussels (Mytilus galloprovincialis) and oysters (Crassostrea gigas) were investigated by liquid chromatography-tandem mass spectrometry in an area on the south coast of Korea from 2007 to 2009. The total lipophilic shellfish toxin (LST) levels in bivalves showed geographical and annual variations. LSTs were detected mostly in the hepatopancreas of mussels from Jinhae Bay throughout the entire year, except in November and December of 2007, but were almost undetectable in all samples during the entire year in 2009. The peak DSP toxin (okadaic acid plus dinophysistoxin 1) levels in the hepatopancreas of mussels from Jinhae Bay and the Tongyeong region were 945.3 and 37.6 ng/g, respectively. The DSP toxin content was about 10 times higher in mussels than in oysters collected from the same region. The major toxins in bivalves were okadaic acid and dinophysistoxin 1; however, pectenotoxin 2 or yessotoxin was occasionally detected as a major component. The results of a quantitative analysis of phytoplankton showed that Dinophysis acuminata was the most probable source of the LSTs, with the exception of yessotoxin. When the highest DSP toxin level was measured (945.3 ng/g in the hepatopancreas of mussels from Jinhae Bay), the toxin concentration in whole mussel tissue was calculated to be 114.0 ng/g. The calculated highest DSP toxin level in whole oyster tissue from both regions was 15.0 ng/g. The calculated maximum toxicities in whole mussel and oyster tissues were lower than the regulatory limit (160 to 200 ng/g) in Korea, the European Union, and the United States. Korean oysters (242 samples) and mussels (214 samples) were thus deemed safe for consumption. But because such variation was detected in a relatively small area of the coast, it is possible that at some locations or during a specific period LST levels could exceed the standard and a few consumers could be at risk of experiencing DSP.

In 2009, a refined HPLC method based on AOAC Official Method 2005.06 was developed and validated for the determination of paralytic shellfish poisoning (PSP) in mussels. A single-laboratory validation study of this method was undertaken here for the analysis of PSP toxins in oysters, cockles, clams, and razor clams. The method was characterized for selectivity, sensitivity, linearity, precision, repeatability, recovery, ruggedness, and uncertainty of measurement. Validation data were utilized to determine method performance characteristics for non-mussel bivalves for all commercially available certified reference toxins, extending the method to dcNEO and dcGTX2,3, where available. A period of parallel testing of oysters, cockles, and clams enabled a comparison of sample toxicities obtained using mouse bioassay (MBA) and HPLC methodologies, although only a very low number of PSP-positive samples were obtained through the United Kingdom official control monitoring program. Results from the MBA and HPLC methods were well-correlated for PSP-negative samples, but the low number of naturally contaminated PSP-positive samples has prevented any comparative statistical assessment of method performance for non-mussels between the two official methods. However, some evidence for potentially significant differences in total saxitoxin equivalents obtained by the two methods in some species has highlighted the need for further comparative testing in non-mussel samples to be conducted prior to implementation of the HPLC method in routine official control monitoring programs.

Okadaic acid, a diarrhetic shellfish poison, domoic acid, an amnesic shellfish poison, and saxitoxin, a paralytic shellfish poison, are three of the best-known marine biotoxins. The mouse bioassay is the method most widely used to detect many of these toxins in shellfish samples, but animal welfare concerns have prompted researchers to seek alternative methods of detection. In this study, three direct competitive enzyme-linked immunosorbent assays (ELISAs), each based on antibodies raised in rabbits against a conjugate of the analyte of interest, were developed for marine biotoxin detection in mussel, oyster, and scallop. One assay was for okadaic acid, one for saxitoxin, and one for domoic acid usually detected and quantified by high-performance liquid chromatography-ultraviolet light (HPLC-UV). All three compounds and a number of related toxins were extracted quickly and simply from the shellfish matrices with a 9 : 1 mixture of ethanol and water before analysis. The detection capabilities (CCbeta values) of the developed ELISAs were 150 microg kg(-1) for okadaic acid, 50 microg kg(-1) for domoic acid, and 5 microg kg(-1) or less for saxitoxin. The assays proved satisfactory when used over a 4-month period for the analysis of 110 real samples collected in Belgium.

Bivalve shellfish samples containing paralytic shellfish poisoning toxins were subjected to gamma irradiation dosage trials in order to assess the potential suitability of the technique in the production of toxin reference materials. Two candidate reference materials of tissue homogenates, mussels (Mytilus sp.) and native oysters (Ostrea edulis), were prepared in-house. Both were subjected to gamma irradiation at four different dose levels, 3.0, 6.0, 13.0 and 18.1 kGy. Bacterial levels were shown to be eliminated in the mussels and significantly reduced in the oysters following irradiation at all four dose levels. Paralytic shellfish poisoning (PSP) toxin concentrations were not significantly reduced in any of the samples indicating the treatment had no adverse affect on the initial stability of any of the PSP toxins monitored. Chromatographic results showed near-identical profiles for treated and non-treated samples inferring that no fluorescent toxin degradation products or matrix interferences were produced during the irradiation process. Results therefore proved that gamma irradiation treatment reduced bacterial levels within paralytic shellfish poisoning reference materials without compromising analyte content, with the subsequent potential to enhance the stability of future candidate reference materials treated in this manner.

The lagoonal system Altata-Ensenada del Pabellón supports important traditional fisheries and mollusc cultures and receives urban and agricultural effluents. The annual mean Cd contents of the oyster and mussel Crassostrea gigas and Mytella strigata of the inner mangrove swamps were higher than that of the clam Megapitaria squalida, which lives in areas under marine influence. Crassostrea corteziensis had the highest Cu and Zn contents, showing that it is a strong accumulator of both metals and especially of Zn, and there were no significant differences in the Pb content of the three species.

With data from the annual analyses of mussels and oysters collected in 1986-1993 from sites located throughout the coastal United States [O'Connor, T.P., 1996. Trends in chemical concentrations in mussels and oysters collected along the US coast from 1986 to 1993. Mar. Environ. Res. 41, 183-200] showed decreasing trends, on a national scale, for chemicals whose use has been banned or has greatly decreased and that concentrations of most other chemicals were neither increasing nor decreasing. With data through 2003 those conclusions still apply. National median concentrations of synthetic organic chemicals and cadmium continue to decrease. The added data show that concentrations of lindane and high molecular weight PAHs are also decreasing on a national scale. For metals other than cadmium and zinc (in mussels), the added data reveal trends at more sites than in 1993 but no additional national trends. However, the longer time series has revealed several local and regional trends.

Bivalve tissue samples were examined over a range of spatial and temporal scales (1993-2001) to determine PAH distributions, trends, and possible sources. Mussels (Mytilus californianus), oysters (Crassostrea gigas), and clams (Corbicula fluminea) were deployed for three months in the estuary at stations remote from known point source discharges. The range of summation operatorPAH detected in bivalves was oysters 184-6899 microg/kg dry wt (mean 678 microg/kg dry wt), mussels 21-1093 microg/kg dry wt (mean 175 microg/kg dry wt), and clams 78-720 microg/kg dry wt (mean 323 microg/kg dry wt). Linear regression analysis showed no statistically significant (p>0.05) temporal trends in clam and mussel summation operatorPAH at any of the deployment stations or estuary segments. On the other hand, a statistically significant (p<0.05) decreasing trend was found in summation operatorPAH in oysters at the Petaluma River station, and in the North Estuary segment. PAH isomer pair ratios applied as diagnostic indicators suggested that the bioaccumulated PAH were derived primarily from petroleum combustion, with lesser amounts derived from biomass and coal combustion, and unburned petroleum.

The bivalves, Anadara (Senilia) senilis (n=95), Crassostrea tulipa (n=75) and Perna perna (n=30) from Ghana, were analyzed for their organochlorines (pesticides and PCBs) concentrations and the body burden. A comparison was done based on two different standard PCB mixtures (Aroclor 1254 and 1260) with the percentage individual congener contribution and that of the sample. From these comparisons, it appeared that the pattern of PCB residues in the samples was neither Aroclor 1254 nor 1260. On the other hand, there was seasonal qualitative difference in the pattern that is purely marine (in the dry season) versus terrestrial input (in the wet season). SigmaPCB was 0.10 microg/g dw or 2.2 microg/g lw (median values). There was no correlation between PCB concentration and lipid content reflecting the importance of indirect contamination of the bivalves. The pesticides, aldrin, dieldrin, endrin, lindane, DDTs, HCH, HCB, endosulfan, heptachlor epoxide and heptachlor, were detected. The most abundant pesticides were the SigmaDDT: 73 and SigmaHCH: 29 ng/g dw. DDT/DDE and hept./heptachlor epoxide ratios reflect their recent applications, while the ratio for aldrin/dieldrin suggests that the use of aldrin has been discontinued, at least on the coast of Ghana.

The Gulf of Paria is bordered by both Trinidad and Venezuela, from which various metallic pollutants and other contaminants can originate. The Gulf is still a significant source of fish, crabs and shellfish for human consumption to both countries, where concerns over the quality of this marine environment have been long expressed but never properly addressed. In addition, the circulatory current patterns in the Gulf ensure that contaminants originating from either country are likely to affect both countries eventually. Heavy metals were determined in oysters (Crassostrea rhizophorae and C. virginica), green mussels (Perna viridis) and sediments from the Gulf of Paria. Samples were obtained at four sites in Trinidad and three sites in Venezuela in the Gulf of Paria, in addition to comparative samples collected from three sites on the north coast of Venezuela. Edible tissues of twelve shellfish from each location were blended and aliquots digested with concentrated nitric acid, for extraction of cadmium, chromium, copper, lead, nickel and zinc. The solutions were analysed by flame atomic absorption spectroscopy. Mercury was extracted with a mixture of nitric, hydrochloric and sulphuric acids and determined by cold vapour atomic absorption. Sediments were oven-dried at 60'C, before being similarly extracted. Results showed that mercury in sediments at all sites in Trinidad and Venezuela exceeded NOAA and Canadian sediment quality guidelines, while cadmium, copper, nickel, lead and zinc also exceeded these guidelines at several sites. Heavy metal levels in oysters and green mussels varied widely with location. However, oysters from the Gulf of Paria contained significantly higher mean levels of cadmium, copper, nickel and zinc than those from the north coast of Venezuela, but this difference was not apparent in mussels. Cadmium, mercury and zinc in sediments were significantly correlated with those of mussels, but not of oysters, in which copper and zinc at several sites in the Gulf of Paria exceeded local maximum permissible levels (Cu = 20 microg g(-1) wet wt; Zn = 50 microg g(-1) wet wt) for human consumption. These findings indicate that while mussels may be better biological indicators of heavy metal pollution in sediments than oysters, the latter may provide copper and zinc contamination. Further research is needed to determine the most appropriate biological indicators of heavy metal and other pollutants in the local marine environment and to develop protocols for their use.

A 15-month experiment combining a geochemical survey of Cd, Cu, Zn and Hg with a bioaccumulation study for three filter-feeding bivalve species (oysters, Crassostrea gigas; cockles, Cerastoderma edule; and clams, Ruditapes philippinarum) was conducted in a breeding basin of the Nord Medoc salt marshes connected to the Gironde estuary, which is affected by historic polymetallic pollution. Regular manual surface measurements of temperature, salinity, pH and dissolved O(2) concentration and hourly multiprobe in situ measurements throughout several periods for 6-8 weeks were performed. The geochemical behavior of metals in water, suspended particulate matter and sediment and their ecotoxicological impact on the three bivalve species were evaluated by in situ exposure of juvenile oysters (water column) and adult cockles and clams (sediment surface). The physico-chemical parameters reflected seasonal variations and basin management. A distinct daily periodicity (except salinity) indicated intense photosynthesis and respiration. In summer, low dissolved O(2) saturations ( approximately 40-50%) occurred in the early morning at 30 cm above the sediment, whereas in depressions, the water column near the sediment surface was suboxic. Cadmium, Zn and Cu concentrations in suspended particulate matter exceeded typical estuarine values and were much higher than the homogeneously distributed concentrations in different depth ranges of the basin sediment. Particles collected in sediment traps showed intermediate metal concentrations close to sediment values. These results suggest trace metal recycling due to reductive dissolution under suboxic conditions at the sediment surface resulting in trace metal release to the water column and adsorption onto suspended particles. Dissolved Cd, Zn and Hg concentrations (e.g. 13-136 ng l(-1); 0.3-25.1 microg l(-1) and 0.5-2.0 ng l(-1), respectively) in the basin corresponded to the concentration range typically observed in the Gironde estuary, except for some maximum values attributed to metal recycling. In contrast, dissolved Cu concentrations (1.08-6.08 microg l(-1)) were mostly higher than typical estuarine values, probably due to recycled Cu complexation by dissolved organic matter. Growth, bioaccumulation rates and kinetics in the whole soft body of the bivalves were analyzed every 40 days. Although Cd bioaccumulation of oysters was lower in the basin than in the estuary during the same period (27,000 ng g(-1), dry weight and 40,000 ng g(-1), respectively) these values are largely above the new human consumption safety level (5000 ng g(-1), dw; European Community, 2002). For cockles and clams, Cd bioaccumulation was lower, reaching 1400 ng g(-1) and 950 ng g(-1), respectively. Similar results were obtained for Zn and Cu suggesting physiological differences between the species and/or differences in the exposure of the organisms due to physico-chemical conditions and metal distribution between dissolved and particulate phases. In contrast, Hg bioaccumulation was highest for cockles reaching bioconcentration factors of approximately 200,000, which even exceeded that of Cd in oysters (50,000) for the same exposition period. Nevertheless, Hg concentrations remained relatively low in the three bivalve species.

Butyl- and phenyltin residues in bivalves were quantified around the coast of Korean peninsula in 1995-1998 and 2001. Butyltin compounds were detected at all the sites surveyed, whereas phenyltins were detected at 41 out of 67 sites in 1995-1998. Tributyltin (TBT) concentrations in bivalves ranged 48-2800 ng Sn/g on dry weight basis. Triphenyltin (TPT) concentrations ranged <3-1820. The mean TPT concentration was 3.5 times less than that of TBT. Spatial distribution of TBT was closely related to boating and dry-docking activities. TPT concentration in bivalves showed significant correlation with TBT. Concentrations of TBT in bivalves, 2001 are not significantly changed compared to those in 1995-1998. However, TPT concentrations in bivalves steeply decreased in 2001. High concentration of TBT and lesser extent TPT and their nation-wide distribution indicate that organotins are one of major organic pollutants in the marine environment of Korea.

In the United States, the detection of paralytic shellfish poisoning (PSP) for regulatory purposes relies on the mouse bioassay (MBA). Using a saxitoxin presence/absence test could reduce animal usage significantly. Three in vitro methods, the RIDASCREEN Saxitoxin kit, MIST Alert, and a 5 h neuroblastoma assay, were evaluated in parallel with the MBA using 106 twice-frozen, acidified extracts from California-grown mussel and oyster tissues. For each assay, a cutoff point was established whereby data below or equal to that point were scored as negative and were assigned a score of zero. Data above the cutoff were considered positive and assigned a score of one. Pearson correlation coefficients were generated. The RIDASCREEN, MIST Alert, and neuroblastoma bioassay correlated to the MBA at 0.849, 0.853, and 0.832 when used for presence/absence detection. These data suggest that a reduction in MBA usage could be achieved in the surveillance of California-grown mussels and oysters for PSP-associated toxins. Correlation data between the in vitro assays, cost comparisons, and the potential for false negatives and false positives were examined. Implications of these methodologies in protecting public health are discussed.

A method was developed for the quantitative determination of total vanadium concentration in mussels via electrothermal atomic absorption spectrometry (ETAAS). After the microwave digestion of the samples, a program using temperatures of 1600 degrees C and 2600 degrees C for ashing and atomization respectively, without any matrix modifiers, allowed us to obtain results that were satisfactory since they agreed closely with certified reference material values. The detection limit was 0.03 mg kg(-1) (dry weight), indicating that the method is suitable for the analysis of mussel samples. This determination was compared with matrix modifiers that have been reported previously. The method was applied to various cultivated and wild mussels from the Galician coast, yielding levels below 1 mg kg(-1) (wet weight).

In the Thau lagoon, despite there being no phosphate limitation policy, the annual mean phosphate concentrations decreased by 90% between 1971 and 1994. The present study indicates a relationship between the decrease of phosphate concentrations and changes in urban, industrial and agricultural activities within the catchment generating phosphorus (P) inputs; and shellfish farming leading to phosphorus storage and export. The relationship (tonnes of P) is: P-PO(4)(3-) in the lagoon waters=1.0124 anthropogenic phosphorus balance--47.62 (r(2) = 0.824, p < 0.000). As population growth is expected in coastal zones, sustainable development requires the need for addressing the role played by human activities and the multi-scale regulation of them. As an element of integrated coastal zone management, the present study provides a regional scale analysis of these aspects.

The FA composition of glycerophospholipid (GPL) classes and subclasses was investigated in whole animals of three marine bivalve mollusks: the Japanese oyster Crassostrea gigas, the blue mussel Mytilus edulis, and the Manila clam Ruditapes philippinarum. Individual organs (gills, mantle, foot, siphon, and muscle) of the Manila clam also were examined. The PS plasmalogen (PSplsm), PE plasmalogen (PEplsm), and PC plasmalogen (PCplsm) subclasses were isolated by HPLC, and their individual FA compositions were examined using GC. Plasmalogen forms of PS and PE, when compared to their respective diacyl forms, were found to be specifically enriched with non-methylene-interrupted (NMI) FA (7,15-22:2, 7,13-22:2, and their precursors) and 20:1 n-11 FA. Such a clear specific association was not found for PCplsm. Interestingly, this trend was most apparent in PSplsm, and the above FA were found to be, respectively, the predominant PUFA and monounsaturated FA in the PSplsm isolated from the three species. This specificity was maintained in all the analyzed organs of the Manila clam but varied in proportions: The highest level of plasmalogens, NMI FA, and 20:1 n-11 was measured in gills and the lowest was in muscle. These results represent the first comprehensive report on a FA composition of the PSplsm subclass isolated from mollusks. The fact that NMI FA and 20:1 n-11, which are thought to be biosynthesized FA, were mainly associated with aminophospholipid plasmalogens (PE and PS) is likely to have a functional significance in bivalve membranes.

Azaspiracid Poisoning (AZP) is a new toxic syndrome that has caused human intoxications throughout Europe following the consumption of mussels (Mytilus edulis), harvested in Ireland. Shellfish intoxication is a consequence of toxin-bearing microalgae in the shellfish food chain, and these studies demonstrated a wide geographic distribution of toxic mussels along the entire western coastal region of Ireland. The first identification of azaspiracids in other bivalve mollusks including oysters (Crassostrea gigas), scallops (Pecten maximus), clams (Tapes phillipinarium), and cockles (Cardium edule) is reported. Importantly, oysters were the only shellfish that accumulated azaspiracids at levels that were comparable with mussels. The highest levels of total azaspiracids (microg/g) recorded to-date were mussels (4.2), oysters (2.45), scallops (0.40), cockles (0.20), and clams (0.61). An examination of the temporal variation of azaspiracid contamination of mussels in a major shellfish production area revealed that, although maximum toxin levels were recorded during the late summer period, significant intoxications were observed at periods when marine dinoflagellate populations were low. Although human intoxications have so far only been associated with mussel consumption, the discovery of significant azaspiracid accumulation in other bivalve mollusks could pose a threat to human health.

The relationship between Zn, Cu, Cd, Pb, and Ni concentrations in soft tissues of mussels and oysters and those in ambient sediments was examined. The study area was the coastal habitats of the northwestern Sea of Japan, which has a broad range of contamination due to urban sewage. Significant increases of all metals, except Ni, were observed in mussel Crenomytilus grayanus at concentrations of easily leachable metals in ambient sediments higher than 2, 100, and 800 microg/g for Cd, Cu, and Zn, respectively. Pb was accumulated by the mussels with no distinct threshold in Pb contamination of sediments. After 2 months, C. grayanus transplanted from a pristine to a contaminated locality increased in Pb up to 12%, and Cu up to 68% of concentrations of these metals in the contaminated local mussels, but Zn and Cd concentrations increased only in the excretory tissue, not in the somatic tissue, or in the soft tissue as a whole. Such regulation of metal accumulation puts limitations on the use of C. grayanus for monitoring of slightly and moderately contaminated localities. On the other hand, the oyster Crassostrea gigas showed definite accumulation of all metals, except Ni, at moderate contamination of ambient sediments. As contamination of sediments increased, increases of Pb, Zn and Cu concentrations in C. gigas slowed, possibly due to physiological control at very high metal concentrations in oyster soft tissue. Thus, the mussel C. grayanus should be used mainly for the monitoring of heavy contaminated localities, and the oyster C. gigas is more suitable as an indicator of low and moderate contamination.

In this paper, a new high-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP-MS) methodology for the analysis of organotin compounds in complex matrices is described. Earlier studies had failed to show baseline resolution between dibutyltin (DBT) and triphenyltin (TPhT). The data presented in this paper show that, by using a different C-18 stationary phase material (Ace C-18) with decreased particle size, baseline resolution of DBT and TPhT can be achieved, with the resultant separation of a third interfering component. In addition, the Ace C-18 stationary phase yields a significant increase in the number of theoretical plates, and, combined with changes in the mobile phase composition, a reduction in run-time by approximately 25%. It is shown that the minor compounds detected are present in the sample and not artefacts of the analytical procedure. The accuracy and precision of the proposed HPLC-ICP-MS method was demonstrated for the determination of TBT in oyster tissue during the BCR "MULSPOT" international interlaboratory certification project.

Concentrations of 8 heavy metals: cadmium (Cd), copper (Cu), zinc (Zn), lead (Pb), nickel (Ni), chromium (Cr), antimony (Sb) and tin (Sn) were examined in 3 species of bivalves ( Perna viridis, Crassostrea rivularis and Ruditapes philippinarum) collected from 25 sites along the Pearl River Delta coastal waters in the South China Sea from July to August 1996. In general, Cd, Cu, Zn and Sn concentrations in the three bivalve species collected from the Estuarine Zone were significantly higher than those collected from the Western and Eastern Zones of the Pearl River Delta, which are related to the existence of various anthropogenic activities in the catchment of the Pearl River Delta. The Western Estuarine Zone is mainly impacted hy Cr, Ni and Cu contamination. In Victoria Harbor, heavy metal contamination is mainly due to Cu and Pb, Cd, Cu and Zn concentrations in oysters were significantly higher than those in mussels and clams. This could be explained by the fact that oysters live mainly in the Estuarine Zone of the Pearl River Delta which receives most of the polluting discharges from the catchment of the Delta. During turbid condition, heavy metals( soluble or adsorbed on suspended particulates) discharged from the Delta are filtered from the water column and subsequently accumulated into the soft body tissues of oysters. Heavy metal concentrations in the three bivalve species were compared with the maximum permissible levels of heavy metals in seafood regulated by the Public Health and Municipal Services Ordinance, Laws of Hong Kong, and it was revealed that Cd and Cr concentrations in the three bivalve species exceeded the upper limits. At certain hotspots in the Delta, the maximum acceptable daily load for Cd was also exceeded.

This paper compares data of 15 individual polycyclic aromatic hydrocarbon (PAH) concentrations from two monitoring programs: the French Rdseau National d'Observation de la qualité du milieu marin (RNO) and the Mussel Watch project of the U.S. National Status and Trends (NST) program. Polycyclic aromatic hydrocarbon measurements in bivalve (mussels and oysters) are made from samples collected at 265 sites along the U.S. coastline and at 97 sites in the French coastal waters. Individual PAH patterns were found strikingly similar between the two countries with higher concentrations for high-molecular-weight (HMW) PAHs. Principal component analysis results for both RNO and NST show the variability to be dominated by just two components with HMW compounds contributing primarily to the first and low-molecular-weight (LMW) compounds to the second. This could imply a separation of petrogenic and pyrolytic sources with the latter being the more important in both nations accounting for the similarity in results.

Heavy metal concentrations were monitored in edible soft tissues of shellfish from Trinidad and Venezuela. Oysters (Crassostrea sp.) and the green mussel (Perna viridis), which is a recently transplanted species to the Caribbean from the Far East, were collected at six locations in Venezuela and five in Trinidad, the latter along the coast line of the Gulf of Paria. Simple and low-cost methods of analysis were optimized and validated using standard reference materials. Cadmium, copper, lead, nickel, and zinc were analyzed by flame atomic absorption spectrometry. Mercury was determined by cold-vapor atomic absorption spectrometry. The present study has confirmed that oysters have a much greater capacity for accumulation of copper and zinc than does green mussel. In addition, concentrations of copper and zinc in oysters (Crassostrea sp.) at many of the sites in the Gulf of Paria exceeded local and international standards, whereas green mussel P. viridis contained generally acceptable levels for human consumption.

Since 1986 the NOAA National Status and Trends (NS&T) Program Mussel Watch has monitored concentrations of trace chemicals in the coastal United States by sampling mussels, oysters, and sediment. The sediment data have been used to define the status or geographic distribution of chemical concentrations (Daskalakis, K. D., & O'Connor, T. P. (1995). Distribution of chemical contamination in coastal and estuarine sediments. Marine Environmental Research 40, 381-398) and the molluscan data have provided an estimate of temporal trends (O'Connor, T. P. (1996). Trends in chemical concentrations in mussels and oysters collected along the US coast from 1986 to 1993. Marine Environmental Research 41,183-200, O'Connor, T. P. (1998). Mussel Watch results from 1986 to 1996. Marine Pollution Bulletin 37, 14-19). This paper centers on chemical concentrations in mollusks at 263 sites around the United States. It provides perspective on concentration ranges and on geographic distributions. For most organic chemicals and lead, concentrations vary in proportion to numbers of people living near a site. For elements, other than lead, high concentrations in mollusks can be due more to natural factors than to human activity. Concentrations of PAHs in tissues of mussels from urban areas are in a range reported to exert biological responses.

A method using high-performance liquid chromatography/inductively coupled plasma mass spectrometry (HPLC/ICP-MS) has been developed to determine inorganic arsenic (arsenite, arsenate) along with organic arsenic compounds (monomethylarsonic acid, dimethylarsinic acid, arsenobetaine, arsenocholine, trimethylarsine oxide, tetramethylarsonium ion and several arsenosugars) in fish, mussel, oyster and marine algae samples. The species were extracted by means of a methanol/water mixture and a dispersion unit in 2 min, with extraction efficiencies ranging from 83 to 107% in the different organisms. Up to 17 different species were determined within 15 min on an anion-exchange column, using a nitric acid gradient and an ion-pairing reagent. As all species are shown in one chromatogram, a clear overview of arsenic distribution patterns in different marine organisms is given. Arsenobetaine is the major compound in marine animals whereas arsenosugars and arsenate are dominant in marine algae. The method was validated with CRM DORM-2 (dogfish muscle). Concentrations were within the certified limits and low detection limits of 8 ng g(-1) (arsenite) to 50 ng g(-1) (arsenate) were obtained., (Copyright 2002 John Wiley & Sons, Ltd.)

Vitamin B12 content of various edible shellfish was determined by both Lactobacillus leichmannii ATCC 7830 microbiological and intrinsic factor-chemiluminescence methods. The values determined by the microbiological method were 1.2-19.8 (M/C ratio) fold greater in the shellfish than the values determined by the chemiluminescence method. Vitamin B12 compounds were purified from most eaten shellfish, oyster (M/C, 1.5), mussel (M/C, 1.2), and short-necked clam (M/C, 2.7), and partially characterized. TLC and HPLC patterns of each red-colored vitamin B12 compound (M/C, 1.0-1.2) purified from these shellfish were identical to those of authentic vitamin B12. Although the higher values in the determination of vitamin B12 by the microbiological method may be due to the occurrence of vitamin B12-substitutive compounds, the edible shellfish would be excellent vitamin B12 sources judging from the values (> or = 6 micrograms/100 g) determined by the chemiluminescence method.

The methylmercury content in two new marine bivalve mollusk tissue Standard Reference Materials (SRMs) has been certified using results of analyses from the National Institute of Standards and Technology (NIST) and two other laboratories. The certified concentrations of methylmercury were established based on the results from four and six different (independent) analytical methods, respectively, for SRM 1566b Oyster Tissue (13.2 +/- 0.7 microg/kg) and SRM 2977 Mussel Tissue (organic contaminants and trace elements) (36.2 +/- 1.7 microg/kg). The certified concentration of methylmercury in SRM 1566b is among the lowest in any certified reference material (CRM).

Background: Peroxynitrite is increasingly proposed as a contributor to defence system in marine bivalve. It can be formed by combination of superoxide and nitric oxide, and can react with tyrosine residues of proteins giving rise to 3-nitrotyrosine., Results: The present article describes a competitive ELISA for the measurement of 3-nitrotyrosine contents of plasma proteins from marine bivalves by means of a monoclonal anti 3-nitrotyrosine antibody mouse IgG., Conclusions: This assay is sensitive enough to determine the amounts of 3-nitrotyrosine in plasma proteins from one animal only. Using the C-ELISA, we have shown that the phagocytosis of zymosan particles increased the 3-nitrotyrosine levels of plasma proteins from mussel M. galloprovincialis and oyster C. gigas 5.8 and 7.5 times respectively.

Owing to their induction by metals metallothioneins (MTs) have been proposed as biomarkers of the metallic contamination of the environment. On the other hand, bivalves are regarded as very convenient bioindicators of the aquatic ecosystems and an extensive literature has been dedicated to their response to metals. Among studies supporting the involvement of MTs in metal detoxification some discrepancy appears due to inter- and intra-specific variations, or to heterogeneous exposure conditions. A lesser number of papers are dealing with the use of metallothionein levels as biomarkers, and sometimes they evidence that natural factors influencing metallothionein synthesis have to be taken into account before final conclusions can been drawn. Moreover, there is still a large number of non-intercalibrated protocols used to quantify amounts of metallothioneins in organisms. As comparisons are necessary to assess the relative abundance of metallothioneins in a studied species, more work has to be completed before such comparisons could be validated. In the present paper we wish to establish the limits of the use of mollusc metallothioneins as a biomarker of aquatic ecosystem contamination by trace metals, using published and recent data as support for our conclusions and perspectives.