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Simple Summary: Microplastics (MPs) are anthropogenic microscopic pieces of plastic in marine sediments and the water column that can originate from two different sources. Primary MPs are industrially produced microbeads, while secondary MPs result from the fragmentation of larger plastic debris by physical, chemical, and biological processes. Although not yet considered a food contaminant, they are readily ingested by marine organisms at several trophic levels, thus entering the food chain and representing a risk to consumers of fishery products. In this study, Donax trunculus (Truncate donax) specimens collected from Class A production areas in the Tyrrhenian Sea (Mediterranean Sea), and therefore destined directly for the final consumer, were analyzed for the presence of MPs. First, all items were morphologically classified and measured, and some of these were chemically identified. Then, the MPs' mean abundance (MA) was calculated, and a risk assessment of human exposure to MPs was carried out. Both the MA of MPs and human exposure to MPs were found to be low. However, given the magnitude of the problem, the collection of further data using standardized methods is essential for a better risk assessment. Microplastics (MPs) (0.1 µm–5 mm particles) have been documented in oceans and seas. Bivalve molluscs (BMs) can accumulate MPs and transfer to humans through the food chain. BMs (especially mussels) are used to assess MPs' contamination, but the genus Donax has not been thoroughly investigated. The aim of this study was to detect and characterize MPs in D. trunculus specimens collected along the Tuscan coast (Italy), and to assess the potential risk for consumers. The samples (~10 g of tissue and intervalval liquid from 35 specimens) were digested using a solution of 10% KOH, subjected to NaCl density separation, and filtered through 5 μm pore-size filters. All items were morphologically classified and measured, and their mean abundance (MA) was calculated. Furthermore, 20% of them were analyzed by Raman spectroscopy and, based on the obtained results, the MA was recalculated (corrected MA) and the annual human exposure was estimated. In the 39 samples analyzed, 85 items fibers (n = 45; 52.94%) and fragments (n = 40; 47.06%) were found. The MA was 0.23 ± 0.17 items/grww. Additionally, 83.33% of the items were confirmed as MPs (polyethylene and polyethylene terephthalate). Based on the correct MA (0.18 MPs/grww), D. trunculus consumers could be exposed to 19.2 MPs/per capita/year. The health risk level of MPs was classified as level III (moderate). [ABSTRACT FROM AUTHOR]

Concentrations of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (DL-PCBs) were investigated in six species of bivalve mollusc purchased on the Polish retail market. The risk to consumers was calculated as a percentage of the tolerable weekly intake (TWI) (2 pg World Health Organization toxic equivalent (WHO-TEQ) kg−1 body weight (b.w.)). Altogether 32 samples were analysed using an isotope dilution technique with high resolution gas chromatography coupled with high resolution mass spectrometry. Low levels of all analysed compounds were found. The range of PCDD/Fs was 0.08–0.37 pg WHO-TEQ g−1 of wet weight (w.w.) and 0.04–0.41 pg WHO-TEQ g−1 w.w. for DL-PCBs. The highest concentrations of all analysed compounds were found in Pacific oysters, at 0.30 pg WHO-TEQ g−1 w.w. for the sum of PCDD/Fs and 0.19 pg WHO-TEQ g−1 for the sum of DL-PCBs. These concentrations were 2–4 times higher than those detected in the other analysed mollusc species. Different species-dependent congener profiles were observed for PCDD/F concentrations, while PCB congener concentration profiles were species independent. The risk to consumers was assessed relating theoretical intakes of PCDD/Fs and DL-PCBs per 25, 50 and 100 g of consumption of molluscs per week to the TWI. Taking into account the low consumption of molluscs in Poland and low concentrations of analysed compounds, neither adults nor children are likely to exceed the TWI by ingestion of food in this category. [ABSTRACT FROM AUTHOR]

Twelve trace elements were studied in four economically important bivalve molluscs (Mediterranean mussel, manila clam, striped venus clam, and truncated wedge clam) from the southern Marmara Sea for seasonal bioaccumulation and human health risk assessment. According to the findings, Fe and Zn were the most accumulative and element concentrations changed considerably (P < 0.05) between seasons and species. Daily, weekly, monthly intake and target hazard quotients of the trace elements were estimated and compared with maximum limits. The results revealed that consuming these bivalve molluscs from the southern Marmara Sea could be considered safe for human consumption. Nevertheless, further monitoring is needed in order to better follow the development in time of elemental pollution due to increasing environmental pressure on the Marmara Sea basin, together with research on other different bioaccumulative organisms. [ABSTRACT FROM AUTHOR]

This study aimed to assess the presence of thermotolerant Campylobacter spp., as one of the most important foodborne zoonotic pathogens, in three shellfish species: mussels (Mytilus galloprovincialis), oysters (Ostrea edulis) and queen scallops (Aequipecten opercularis). The samples were collected from nine locations in the Istrian aquatory, Croatia. Isolation of Campylobacter was done according to standard ISO method, and species were identified using multiplex PCR. Isolates identified as C. jejuni and C. lari were genotyped using multilocus sequence typing (MLST) to determine the potential source of contamination. Among 108 examined samples of bivalve molluscs, mussels dominated and were the only ones found positive for the presence of Campylobacter (25.6%). In total, 19 C. lari and 1 C. jejuni strains were isolated. C. lari isolates found in this study belong to 13 sequence types (STs), and 9 of them are newly described in this paper. Two out of the four previously described C. lari STs that were found in this study were previously found in human stool. The only C. jejuni isolate was found to be sequence type 1268, which belongs to ST-1275 clonal complex that is almost exclusively found in seabirds and can sporadically cause infection in humans. Regarding the obtained results, introducing surveillance of thermotolerant Campylobacter in shellfish in the Republic of Croatia is advised as an improvement for public health safety. [ABSTRACT FROM AUTHOR]

This work describes a method for the determination of paralytic shellfish toxins in mussels (M. galloprovincialis) , oysters (Crassostrea gigas), and clams (Ruditapes decussatus) samples. The acetic acid extraction method is used for simultaneous analysis of Paralytic Shellfish Poisoning (PSP) and the separation of these compounds was achieved on ethylene bridged hybrid (BEH) Amide column with gradient mobile phase composed of acetonitrile (MeCN) and water (as eluent A and eluent B) both containing 2 mM ammonium formate and 50 mM formic acid. The method specificity, linearity, sensitivity, precision, and accuracy were successfully evaluated towards the detection of 13 hydrophilic marine biotoxins analyzed together (single run) in different sample matrixes such as mussels. oysters. and clams. Calibration curves were found to be linear (R2 ≥ 0.98). average Recovery% of 98.5–99.9%. limits of detection (LODs): 0.16–1.92 µg·kg−1; limits of quantitation (LOQs): 0.54–6.41 µg·kg−1) and the relative standard deviation (RSD%) of the precision was ≤ 2.8%. The method was applied for the determination of PSP in shellfish collected from several Tunisian locations. All LC-MS/MS results of analyzed molluscs collected from different Tunisia locations, reveal no PSP toxin group contamination and that the amount found does not exceeding the threshold of 800 µg STXequ·kg−1. [Display omitted] • A proposed official alternative method of analysis for regulatory control and food safety. • Simultaneous determination of paralytic shellfish toxins using LC-MSMS. • A highly sensitive detection of shellfish biotoxins. • Validation of the method for paralytic shellfish toxins determination. • Effectiveness of proficiency testing within 34 different laboratories. [ABSTRACT FROM AUTHOR]

We examined the occurrence and levels of 19 legacy and emerging per- and polyfluoroalkyl substances (PFASs) in 7 species of marine bivalve molluscs collected from four coastal cities of Shandong Province, China. Perfluorooctanoic acid (PFOA) was the most prevalent component, accounting for 68.1 % of total PFASs. The total PFASs in bivalve molluscs ranged from 0.86 to 6.55 ng/g wet weight, with the highest concentration found in Meretrix meretrix L. The concentration of total PFASs in bivalve molluscs showed the following trend: clams > scallops > oysters > mussels. Estimation on the human intake of PFASs from consumption of bivalve molluscs resulted in hazard ratios (HR) ranging from 0.12 to 6.40. Five of the seven species had HR >1, indicating high exposure risks associated with PFASs. Therefore, the occurrence of PFASs in marine biota is particularly concerning and further investigations on the sources of PFASs in Shandong are warranted. • First time detection of 19 PFASs in bivalve molluscs in Shandong Province • PFASs levels in bivalve molluscs varied from 0.86 to 6.55 ng/g wet weight. • PFOA, PFBA, PFNA and PFOS were the compounds most frequently detected. • Frequent consumption of bivalve molluscs may pose health risk. [ABSTRACT FROM AUTHOR]

Contaminants of Emerging Concerns (CECs) are defined as chemicals not commonly monitored in aquatic ecosystems, but with the potential to cause adverse effects on biota. CECs include Endocrine Disrupting Chemicals (EDCs) and Neuro-Endocrine disruptors (NEDs) of vertebrates. However, most invertebrates only rely on neuroendocrine systems to maintain homeostatic processes. Although conserved neuroendocrine components have been characterized in ecologically relevant groups, limited knowledge on invertebrate neuroendocrinology makes it difficult to define EDCs and NEDs in most species. The monoamine serotonin (5-hydroxytryptamine, 5-HT) acts both as a neurotransmitter and as a peripheral hormone in mammals. In molluscs, 5-HT is involved in multiple physiological roles and molecular components of the serotonergic system have been identified. This review is focused on the effects of CECs on the serotonergic system of bivalve molluscs. Bivalves are widespread in all aquatic environments, estuarine and coastal areas in particular, where they are exposed to a variety of chemicals. In bivalves, 5-HT is involved in gametogenesis and spawning, oocyte maturation and sperm motility, regulates heart function, gill ciliary beating, mantle/siphon function, the "catch" state of smooth muscle and immune responses. Components of 5-HT transduction (receptors and signaling pathways) are being identified in several bivalve species. Different CECs have been shown to affect bivalve serotonergic system. This particularly applies to antidepressants, among the most commonly detected human pharmaceuticals in the aquatic environment. In particular, selective serotonin reuptake inhibitors (SSRIs) are frequently detected in seawater and in bivalve tissues. Information available on the effects and mechanisms of action of SSRIs on the serotonergic system of adult bivalves is summarized. Data are also reported on the effects of CECs on development of neuroendocrine pathways of early larval stages, in particular on the effects of model EDCs in the marine mussel Mytilus galloprovincialis. Overall, available data point at the serotonergic system as a sensitive target for neuroendocrine disruption in bivalves. The results contribute drawing Adverse Outcome Pathways (AOPs) for model EDCs and SSRIs in larvae and adults. However, basic research on neuroendocrine signaling is still needed to evaluate the potential impact of neuroendocrine disruptors in key invertebrate groups of aquatic ecosystems. [ABSTRACT FROM AUTHOR]

Marine biotoxins are toxic substances that may cause illness and death in marine organisms and humans. This article disseminates the results of a 4-year study on the occurrence of marine biotoxins in raw bivalve molluscs purchased from Polish suppliers. A total of 256 samples of 8 different molluscs species were analysed for the presence of biotoxins using the ELISA method for paralytic shellfish poison, diarrhoetic shellfish poison, and amnaesic shellfish poison. The permitted limits of marine biotoxin content were not exceeded in any of the analysed samples and the majority of them were free from these compounds. The results of the study indicate that the tested raw bivalve molluscs available in Poland were safe for consumers. [ABSTRACT FROM AUTHOR]

Highly populated coastal environments receive large quantities of treated and untreated wastewater from human and industrial sources. Bivalve molluscs accumulate and retain contaminants, and their analysis provides evidence of past contamination. Rivers and precipitation are major routes of bacteriological pollution from surface or sub-surface runoff flowing into coastal areas. However, relationships between runoff, precipitation, and bacterial contamination are site-specific and dependent on the physiographical characteristics of each catchment. In this work, we evaluated the influence of precipitation and river discharge on molluscs' Escherichia coli concentrations at three sites in Central Italy, aiming at quantifying how hydrometeorological conditions affect bacteriological contamination of selected bivalve production areas. Rank-order correlation analysis indicated a stronger association between E. coli concentrations and the modelled Pescara River discharge maxima (r= 0.69) than between E. coli concentration and rainfall maxima (r= 0.35). Discharge peaks from the Pescara River caused an increase in E. coli concentration in bivalves in 87% of cases, provided that the runoff peak occurred 1-6 days prior to the sampling date. Precipitation in coastal area was linked to almost 60% of cases of E. coli high concentrations and may enhance bacterial transportation offshore, when associated with a larger-scale weather system, which causes overflow occurrence. [ABSTRACT FROM AUTHOR]

Bivalve molluscs possess effective cellular and humoral defence mechanisms against bacterial infection. Although the immune responses of mussels to challenge with pathogenic vibrios have been largely investigated, the effects at the site of injection at the tissue level have not been so far evaluated. To this aim, mussels Mytilus galloprovincialis were herein in vivo challenged with Vibrio splendidus to assess the responses induced in hemolymph and posterior adductor muscle (PAM), being the site of bacterial infection. The number of living intra-hemocyte bacteria increased after the first hour post-injection (p.i.), suggesting the occurrence of an intense phagocytosis, while clearance was observed within 24 h p.i. A recruitment of hemocytes at the injection site was found in mussel PAM, together with marked morphological changes in the volume of muscular fibers, with a recovery of muscle tissue organization after 48 h p.i. A concomitant impairment in the osmoregulatory processes were observed in PAM by an initial inhibition of aquaporins and increased immunopositivity of Na+/K+ ATPase ionic pump, strictly related to the histological alterations and hemocyte infiltration detected in PAM. Accordingly, an intense cell turnover activity was also recorded following the infection event. Overall, results indicated the hemolymph as the system responsible for the physiological adaptations in mussels to stressful factors, such as pathogenicity, for the maintenance of homeostasis and immune defence. Also, the osmotic balance and cell turnover can be used as objective diagnostic criteria to evaluate the physiological state of mussels following bacterial infection, which may be relevant in aquaculture and biomonitoring studies. Unlabelled Image • Mussels Mytilus galloprovincialis were in vivo challenged with Vibrio splendidus. • The number of living intra-hemocyte bacteria increased after 1 h post-infection. • Histological changes and recruitment of hemocytes occurred at the injection site. • Impairment of osmoregulation and cell turnover activity were found in mussel muscles. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Natural radionuclides and 137Cs in different types of bivalve molluscs were determined by alpha and gamma spectrometry. • 137Cs was below the minimum detectable activity concentration in all bivalves samples. • The annual ingestion doses due to bivalves ingestion were assessed. • A statistical analysis of natural radioactivity was used to discriminate between molluscs of Atlantic and Mediterranean. Abstract The presence of natural and artificial radionuclides in the marine environment produces the accumulation of radionuclides in bivalve molluscs consumed by humans, and therefore it could result in a radiological hazard. In this study, the activity concentrations of 210Po, 40K, 210Pb and 234Th were determined in different types of bivalve molluscs sampled during the period of May 2014–June 2015, along coastal areas from the Andalusian region (South of Spain), through alpha-particle spectrometry and low-level gamma-ray spectrometry. The activity concentrations of 210Po; 40K; 210Pb and 234Th varied between 40 ± 2 and 515 ± 9 Bq kg−1 dry weight (d.w.); 121 ± 7 and 674 ± 34 Bq kg−1 d.w.; ND (lower than limit of detection) and 73 ± 10 Bq kg−1 d.w.; and ND and 126 ± 27 Bq kg−1 d.w., respectively. The committed effective dose to humans was calculated to range from 41 to 479 μSv year−1. Both activity concentrations and dose levels were comparable to previous studies from other countries. Finally, a multivariate statistical analysis of natural radioactivity content allowed the discrimination between bivalve molluscs from Atlantic and Mediterranean coasts. [ABSTRACT FROM AUTHOR]

Abstract Worldwide bivalve production has increased significantly in recent years. Bivalve meat is recommended in dietary regimens due to the benefits associated with their consumption but, at the same time, is internationally recognized as a potential vehicle for outbreaks of foodborne disease. To reduce food risk exposure, consumers must pay close attention when purchasing, handling and consuming bivalves. This study aimed to collect information on consumers' perceptions and practices related to bivalve purchasing, managing and consumption at home. A focus group methodology was adopted to better understand consumers' food practices and risk perceptions regarding molluscs. Four focus groups involving 42 consumers were conducted in Veneto and Marche region. The core analysis was then applied to each focus group discussion in order to identify, categorize and rank the information collected. The most important factors that came to light during the focus groups concerned the personal reasons for preparing bivalves at home; the place of purchase; the elements guiding consumers when purchasing bivalves; the bivalve preparation, cooking and storage conditions at home; the consumption of raw molluscs and the health risks; the consumers' categories most at risk; and the production chain control by the authorities. The data reported in the article can provide useful information to public authorities working on food safety for developing effective risk communication strategies. Highlights • Consumers play a crucial role in food risk reduction. • The focus group methodology collected information from Italian consumers. • The study analysed consumers' purchasing, handling and consumption of bivalves. • The data provide useful information for implementing risk communication activities. [ABSTRACT FROM AUTHOR]

Highlights • Bivalve molluscs species were analyzed by ICP OES and ICP-MS. • Most Lucina pectinata samples had higher concentrations of Cu, Cr and Pb. • Concentrations of As obtained in most samples were above the legal limit recommended by ANVISA. • PCA and HCA showed separation between Lucina pectinata and Trachycardium muricatum species. Abstract Four species of bivalve molluscs (Anomalocardia brasiliana , Iphigenia brasiliana , Lucina pectinata and Trachycardium muricatum) were collected in the Todos os Santos Bay (TSB), Bahia, Brazil, in order to evaluate As, Cd, Co, Cu, Cr, Fe, Mn, Ni, Pb, Se, V and Zn levels and, consequently, the risk of bivalve mollusc consumption in humans. The samples were analyzed by inductively coupled plasma optical emission spectrometry (ICP OES) and inductively coupled plasma mass spectrometry (ICP-MS) after closed-vessel microwave digestion. The accuracy was confirmed using the certified reference materials of oyster tissue (NIST 1566b) and mussel tissue (NIST 2977), and the results were statistically equivalent to the certified values. Application of principal component analysis (PCA) and hierarchical cluster analysis (HCA) showed a tendency to form two groups between samples of Lucina pectinata and Trachycardium muricatum. All species showed As and Cr concentrations higher than the maximum tolerable limit specified in Brazilian legislation. [ABSTRACT FROM AUTHOR]

Norovirus (NoV) and Hepatitis A virus (HAV) are the most important viral hazards associated with human illness following consumption of contaminated bivalve molluscs. The effectiveness of the current EU criteria for heat processing of bivalve molluscs (i.e. raising the temperature of the internal mollusc flesh to at least 90°C for a minimum of 90 seconds) was evaluated using predictive microbiology. A HAV thermal inactivation model was developed based on literature data in mollusc matrices during isothermal heat treatment. Application of the developed model demonstrated that the 90°C-90 s requirement may lead to significantly different virus inactivation depending on the commercial process design. This shows the need for the establishment of a Performance Criterion for bivalve molluscs heat processing which will assure a common specified level of consumer protection. A risk-based approach is described that allows for an effective processing design providing a more transparent and objective relation between the thermal processing targets and public health. Model simulations demonstrate that the F-value is a more appropriate Process Criterion than a single time-temperature combination since it enables the food business operators to design a process that is compliant with the safety requirements while at the same time achieving a desired product quality. [ABSTRACT FROM AUTHOR]

Bivalve molluscs, including oysters and scallops, are hedonic foods that have been a valuable source of vitamins, minerals and proteins in the human diet since the Stone Age. Despite being rich in beneficial compounds, it is still debatable whether bivalves should be regarded as a healthy food since they can accumulate harmful microorganisms, and toxic compounds from the surrounding environment. Currently, there is still no in-depth understanding of the ultimate effect of bivalve consumption on humans. The use of advanced foodomics approaches is beginning to provide a more detailed characterisation of the beneficial and deleterious compounds in these seafood products. In addition to defining bivalve molluscs within an international dietary framework, this review highlights the major nutritional components of their metabolome (foodome) with a focus on several groups of toxicants whose presence can negatively offset their nutritional value. An overview of metabolomics applications to the study of bivalve molluscs is included. [ABSTRACT FROM AUTHOR]

The Toll/interleukin-1 receptor (TIR) domain has a fundamental role in the innate defence response of plants, vertebrate and invertebrate animals. Mostly found in the cytosolic side of membrane-bound receptor proteins, it mediates the intracellular signalling upon pathogen recognition via heterotypic interactions. Although a number of TIR-domain-containing (TIR-DC) proteins have been characterized in vertebrates, their evolutionary relationships and functional role in protostomes are still largely unknown. Due to the high abundance and diversity of TIR-DC proteins in bivalve molluscs, we investigated this class of marine invertebrates as a case study. The analysis of the available genomic and transcriptomic data allowed the identification of over 400 full-length sequences and their classification in protein families based on sequence homology and domain organization. In addition to TLRs and MyD88 adaptors, bivalves possess a surprisingly large repertoire of intracellular TIR-DC proteins, which are conserved across a broad range of metazoan taxa. Overall, we report the expansion and diversification of TIR-DC proteins in several invertebrate lineages and the identification of many novel protein families possibly involved in both immune-related signalling and embryonic development. [ABSTRACT FROM AUTHOR]

BACKGROUND The increase in seafood consumption and the presence of different species of bivalves on the global markets has given rise to several commercial frauds based on species substitution. To prevent and detect wilful or unintentional frauds, reliable and rapid techniques are required to identify seafood species in different products. In the present work, a pyrosequencing-based technology has been used for the molecular identification of bivalve species. RESULTS Processed and unprocessed samples of 15 species belonging to the bivalve families Pectinidae, Mytilidae, Donacidae, Ostreidae, Pharide and Veneridae were analysed and correctly identified by the developed pyrosequencing-based method according to the homology between query sequences of the 16S ribosomal RNA (16S rRNA) and cytochrome c oxidase I (COI) genes and their correspondent reference libraries. This technique exhibits great potential in automated and high-throughput processing systems, allowing the simultaneous analysis of 96 samples in shorter execution and turnaround times. CONCLUSIONS The correct identification of all the species shows how useful this technique may prove to differentiate species from different products, providing an alternative, simple, rapid and economical tool to detect seafood substitution frauds. © 2016 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Endocrine disrupting chemicals (EDCs) in estuaries and coastal habitats have been widely detected over the world and caused global concern. Bivalves have been shown to be vulnerable to endocrine disruption. However, estrogen receptors (ERs) sensitivity to steroids and EDCs has long been considered to be restricted to vertebrates. In the present study, a computational simulation docking model was applied to qualitatively predict the binding behavior of two bivalve ERs to estradiol and compared the docking activity with zebra fish ER a. A novel reconstituted yeast system was constructed by using transcriptional activator GAL-4 consists of ER-expressing plasmid and ERE (estrogen responsive element)-containing plasmid. The assays showed that bivalve ER specifically activate transcription in response to tested steroids and EDCs, but the activation ability is weaker compared to zebra fish ER a. The results corroborate the presence of an active ER in bivalve molluscs and provide a promising tool for screening of marine environmental pollutants active in disturbing ERs of bivalves, as well as understanding the underlying mechanism across taxonomic groups and phyla. [Display omitted] • Two key amino acid residues of bivalve ER play a role in forming hydrogen bonds with estradiol. • Bivalve ERs specifically activate transcription in response to tested steroids and EDCs. • The response pattern of ERs was comparable among the two bivalves and zebrafish. [ABSTRACT FROM AUTHOR]

Continuous European Union programmes with specified methods for enumeration of Escherichia coli in bivalves for human consumption are currently running. The objective of this research was to examine the species accuracy of the five times three tube Most Probable Number (MPN) EU reference method used for detection of E. coli in marine bivalves. Among 549 samples of bivalves harvested from Norwegian localities during 2014 and 2015, a total number of 200 bacterial isolates were prepared from randomly selected culture-positive bivalves. These presumptive E. coli isolates were characterized biochemically by the Analytical Profile Index (API) 20E, as well as by Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS). The majority of isolates (90%) were identified as E. coli , by both API 20E and MALDI-TOF MS. Ten isolates (5%) were identified as Klebsiella pneumoniae , while one isolate was identified as K. oxytoca by both methods, whereas three isolates were identified as Acinetobacter baumannii , Citrobacter braakii , and Enterobacter cloacae , respectively. The identification of the remaining six isolates were not in compliance between the two methods. [ABSTRACT FROM AUTHOR]

The difficulty in supplying the animal protein demand of great biological value for the human population is a major concern of our century. According to the FAO, the fishery (extractive fishing and aquaculture), will be one of the main activities responsible for food safety in the world, with animal protein production forecasts for the next decade longer than the slaughter of animals sector (beef, pork and poultry). The use of fishing resources with underutilized potential for exploitat ion can ensure the maintenance of primary supply of raw materials on-site domestic and / or regional, and inhibit movement of prices, as the practice of shellfish. In this context, the present review aims to highlight the potential of Anomalocardia brasiliana to exploitation and processing contributing to the strengthening of this production chain in the craft segment and product diversification in the Brazilian market. [ABSTRACT FROM AUTHOR]

Reactive oxygen and nitrogen species (ROS and RNS) are naturally produced in all cells and organisms. Modifications of standard conditions alter reactive species generation and may result in oxidative stress. Because of the degradation of marine ecosystems, massive aquaculture productions, global change and pathogenic infections, oxidative stress is highly prevalent in marine bivalve molluscs. Haemocytes of bivalve molluscs produce ROS and RNS as part of their basal metabolism as well as in response to endogenous and exogenous stimuli. However, sources and pathways of reactive species production are currently poorly deciphered in marine bivalves, potentially leading to misinterpretations. Although sources and pathways of ROS and RNS productions are highly conserved between vertebrates and invertebrates, some uncommon pathways seem to only exist in marine bivalves. To understand the biology and pathobiology of ROS and RNS in haemocytes of marine bivalves, it is necessary to characterise their sources and pathways of production. The aims of the present review are to discuss the currently known and unknown intracellular sources of reactive oxygen and nitrogen species in marine bivalve molluscs, in light of terrestrial vertebrates, and to expose principal pitfalls usually encountered. [ABSTRACT FROM AUTHOR]

Algal biotoxins, chemical compounds produced by some microscopic algae, constitute the phytoplankton. The mussels, feeding on phytoplankton, can accumulate these compounds to become themselves toxic. There have been several cases of food poisoning by consumption of contaminated shellfish. Such food poisoning have pushed our health care system to provide monitoring of shellfish in the framework of the monitoring plans carried out by AASSLL. In this paper we report the results obtained monitoring the presence of amnesic shellfish poisoning biotoxins, like domoic acid and its isomers, produced by Pseudonitzschia algae. The analyses were carried out by using both the high-performance liquid chromatography-ultraviolet official method and an experimental method performed with a time-off-light mass spectrometer (ESI-TOF). The 100% of samples analysed by the official method have always been below the limits of sensitivity (except one sample), and the 65% of samples analysed by ESI-TOF showed the presence of domoic acid. [ABSTRACT FROM AUTHOR]

The input of anthropogenic contaminants to the aquatic environment is a major concern for scientists, regulators and the public. This is especially relevant in areas such as the Tamar valley in SW England, which has a legacy of contamination from industrial activity in the nineteenth and twentieth centuries. Following on from previous laboratory validation studies, this study aimed to assess the relationship between genotoxic and cytotoxic responses and heavy metal concentrations in two bivalve species sampled from locations along the Tamar estuary. Adult cockles, Cerastoderma edule, and blue mussels, Mytilus edulis, were sampled from five locations in the Tamar and one reference location on the south Devon coast. Bivalve haemocytes were processed for comet and neutral red retention (NRR) assays to determine potential genotoxic and cytotoxic effects, respectively. Sediment and soft tissue samples were analysed for metal content by inductively coupled plasma mass spectrometry. Sediment concentrations were consistent with the physico-chemical nature of the Tamar estuary. A significant correlation ( P = 0.05) was found between total metal concentration in sediment and C. edule soft tissues, but no such correlation was found for M. edulis samples. DNA damage was elevated at the site with highest Cr concentrations for M. edulis and at the site with highest Ni and Pb concentrations for C. edule. Analysis of NRR revealed a slight increase in retention time at one site, in contrast to comet data. We conclude that the comet assay is a reliable indicator of genotoxic damage in the field for both M. edulis and C. edule and discuss reasons for the apparent discrepancy with NRR. [ABSTRACT FROM AUTHOR]

Abstract: During two surveys conducted in 2008 and 2009, the culture method described in the international standard ISO/TS 21872–1 was applied to the detection of Vibrio parahaemolyticus and Vibrio cholerae in 112 living bivalve mollusc samples, with a chromogenic medium used in addition to the TCBS agar, as second selective isolation medium and for enumeration of V. parahaemolyticus and V. cholerae by surface inoculation. A PCR method for detection of these 2 Vibrio species and the hemolysin genes tdh and trh, was applied in parallel. In 2009, the survey was extended to finfish fillets and crustaceans. PCR was also used for species confirmation of characteristic colonies. The identity of the PCR products, specifically targeting V. parahaemolyticus, was checked by sequencing. Occurrence of V. parahaemolyticus and V. cholerae isolates in living bivalve molluscs ranged from 30.4% to 32.6% and from 1.4% to 4.7% respectively. In frozen crustaceans (2009 survey) V. parahaemolyticus and V. cholerae isolates were respectively found in 45% and 10% of the samples. No V. parahaemolyticus or V. cholerae was detected in frozen fish fillets, neither by the ISO method nor by PCR. In 2009, enteropathogenic V. parahaemolyticus (trh+) was isolated from 4 out of 43 oyster samples while the trh gene was present in V. alginolyticus strains and in samples where V. parahaemolyticus was not detected (9 over 112 samples). The ISO method failed to isolate V. parahaemolyticus in 44% to 53% of the living bivalve molluscs where PCR detected the toxR gene specific of V. parahaemolyticus (Vp-toxR). Our results highlighted the need for a revision of the ISO/TS 21872–1 standard, at least, for analysis of living bivalve molluscs, and confirmed the increasing concern of enteropathogenic V. parahaemolyticus in French bivalve molluscs. Enrichment at 41.5°C was questioned and some reliable solutions for the improvement of the ISO/TS 21872–1 method, such as the PCR method for screening of positive samples and confirmation of colonies, were pointed out. [Copyright &y& Elsevier]

Abstract: The concentration of Cd, Cr and Pb in the following species Ameghinomya antiqua, Aulacomya atra and Mytilus chilensis, bivalves from Iloca, Constitución and Pelluhue, coastal towns located in the Maule Region, Chile was determined. Representative tissue samples of each species were analysed by using atomic absorption spectroscopy with flame. Validation of methodology was carried out using a certified reference material (TORT-1). The ranges of concentrations found for Cd, Cr and Pb in the 3 species were, respectively, 0.21–4.32, 0.38–12.62 and 0.43–31.10mgkg−1 dry weight. In relation to the results obtained and the average of each individual metal in the different species of bivalves, Pb concentrations were found in higher levels rather than those of other metals reported by other authors in a similar work, this element constitutes a potential health hazard for the consumers of shellfish. The results were statistically evaluated for possible correlations in the content of metals in different species and in different sites of origin. [Copyright &y& Elsevier]

The article presents a study on the estradiol-17ß and testosterone levels in the cockle Fulvia mutica during the annual reproductive cycle in China. The study underwent investigation on seasonal variations in the concentrations of estradiol-17ß and testosterone in the gonad of the cockle Fulvia mutica from March 2004 to February 2005 relative to the reproductive cycle. Histological data revealed that Fulvia mutica is a hermaphrodite bivalve specie and development and maturation of sex cells took place during winter and spring and spawning happened in summer when water temperature was higher.

Abstract: Metallothionein (MT) and other biomarker levels were measured in Scrobicularia plana clams to assess pollution of the Guadalquivir Estuary possibly affected by metals released from Aznalcóllar pyrite mine in 1998. After optimizing reagent concentrations for monobromobimane derivatization, MT levels were quantified by reversed-phase high-performance liquid chromatography coupled to fluorescence detection (RP-HPLC-FD) in heated or unheated digestive gland extracts and compared to those obtained by differential pulse polarography (DPP). MT content assayed by RP-HPLC-FD in unheated samples was higher than that obtained by DPP and correlated better with metals and anti-oxidant activities. MT assay by RP-HPLC-FD in unheated extracts would be preferable for assessing metal pollution, due to its greater sensitivity and specificity. In addition to MT induction, glyoxalase II inhibition was well correlated with metal contents. Our results suggest that metals at the estuary do not originate from Aznalcóllar spill, but from those carried along by the river and deposited at its concave bank. [Copyright &y& Elsevier]

In this work data of salinity, temperature, suspended particulate matter (SPM), chlorophyll-a, and phytoplankton concentration in the Gibraltar Strait coast, in the confluence of the Mediterranean Sean and the Atlantic Ocean, were analyzed together with 210Po, 40K, 210Pb and 234Th activity concentration in different types of bivalve molluscs at the same time-period. The physicochemical parameters were evaluated using the Copernicus Marine Environment Monitoring Service (CMEMS) products based on satellite observations. A multivariate statistical analysis, including contrasted natural radioactivity contents, allowed the discrimination of bivalve molluscs from Atlantic and Mediterranean coasts. Additionally, a cluster analysis determined a highly significant negative correlation 210Po concentration in molluscs and phytoplankton concentration suggesting that phytoplankton concentration in the water column is a determinant factor to regulate 210Po concentration in those animals. These results introduce a useful tool to calculate the radiological doses in seafood from chlorophyll satellite image. • First time that physicochemical parameters based on satellite observations were analyzed with activity concentration. • A significant negative correlation was shown between 210Po concentration in molluscs and phytoplankton concentration. • A multivariate statistical analysis allowed the discrimination of bivalve molluscs from Atlantic and Mediterranean coasts. • Radiological doses in seafood were obtained through chlorophyll satellite image. [ABSTRACT FROM AUTHOR]

Abstract: In the framework of the national monitoring program for marine biotoxins in bivalve molluscs for 2002 and 2003, a total of 862 samples were examined to investigate the presence of domoic acid (DA). Samples mainly consisted of mussels (Mytilus galloprovincialis) and venus clams (Venus verucosa) from various Greek harvesting areas. In 2002, 83% of all samples examined contained less that 1μg DA/g, while in 2003 this percentage was 95%. In both years, DA was only detected in springtime without any sample exceeding the regulatory level. In 2002, the highest value was 14.0μg/g in mussels, while in 2003 these were 4.2μg/g in mussels and 5.6μg/g in venus clams. This is the first report regarding the presence of ASP toxins in Greek shellfish. [Copyright &y& Elsevier]

Abstract: Trace metals were determined in the two most abundant species of bivalve molluscs along the Atlantic coast of southern Spain (Donax trunculus and Chamelea gallina) and in the sediments where they live. The results show that the area near the mouth of the Huelva estuary is where the highest metal concentrations are found in sediments and in the two bivalve species. This is not surprising, considering that the Huelva estuary is the mouth of the Tinto and Odiel rivers, which have one of the highest levels of metal pollution of all the rivers of Europe. The two species of bivalves have different amounts of metals in their tissues. The concentrations of Cr, Cu, Pb, Zn, As and Hg in D. trunculus were significantly higher (p &#60;0.05) than in C. gallina; however, C. gallina contained more Ni and Cd. In both species the most abundant elements were Cu and Zn, while Hg showed the lowest values. There is a significant correlation (p &#60;0.05) for concentrations of Cu, Pb, Zn and Hg in D. trunculus and C. gallina relative to their concentrations in surface sediments. [Copyright &y& Elsevier]

The current study presents data on the antimicrobial resistance (AMR) patterns of 102 S. enterica subsp. enterica (72 Salmonella ser. Typhimurium and 30 monophasic S. Typhimurium serovar) and 79 Escherichia coli (and their phylogenetic group determination) isolates from different species of bivalve molluscs and from the water samples collected from the sub-areas of a mollusc production area near Ferrara (Italy). These areas were classified as Long-line, Lupini, B-Out, B-in, and Sacca. A retrospective evaluation was performed to assess the spatial trends of the resistance patterns of Salmonella and E. coli and the temporal trend for Salmonella ; the role of molluscs as AMR indicators and the potential use of E. coli as a microorganism indicator of AMR occurrence in a seawater environment were also investigated. Overall, 81% of Salmonella spp. and 75% of E. coli isolates were resistant to, at least, one antimicrobial agent (AA) and 44% and 38% of the isolates were multidrug resistant (MDR), respectively. The resistance levels of Salmonella were influenced by the investigated serovars. Monophasic S. Typhimurium serovar showed the highest resistance value with 70% of MDR isolates, in contrast with only 33% in S. Typhimurium. In monophasic S. Typhimurium versus S. Typhimurium, twofold resistance levels were observed to streptomycin (97 versus 43%), ampicillin (80 versus 40%) and tetracyclines (67 versus 36%). Regarding the temporal trend for Salmonella , strains were resistant to, at least, one AA, but this resistance was significantly lower during the first years of this 17-year sampling; however, in parallel MDR isolates, the resistance increased from 23% to a maximum level of 57% during the 2008–2012 period. On assessing the spatial trends, the Sacca area was found to show the lowest number of Salmonella spp. and E. coli strains resistant to, at least, one AA and MDR. For E. coli , the most commonly observed resistance was towards ampicillin (56%), streptomycin (52%), sulphonamides (30%) and ceftiofur (24%). The great majority (65%) of E. coli isolates belonged to the commensal phylogroups A and B1, with B1 as the dominant one, whereas most MDR isolates belonged to phylogroup C. Molluscs may be an efficient tool for antimicrobial resistance monitoring, and E. coli could be used as a microorganism indicator of the occurrence of antimicrobial resistance in seawater environment. • Bivalve molluscs are confirmed as efficient tool for antimicrobial resistance monitoring. • High multi-resistant levels were observed in Salmonella spp. and E. coli from molluscs and water. • Resistance levels for Salmonella are influenced by the investigated serovars. • Spatial trends of MDR Salmonella strains are observed. [ABSTRACT FROM AUTHOR]

Klebsiella spp. are a major cause of both nosocomial and community acquired infections, with K. pneumoniae being responsible for most human infections. Although Klebsiella spp. are present in a variety of environments, their distribution in the sea and the associated antibiotic resistance is largely unknown. In order to examine prevalence of K. pneumoniae and related species in the marine environment, we sampled 476 batches of marine bivalve molluscs collected along the Norwegian coast. From these samples, K. pneumoniae (n = 78), K. oxytoca (n = 41), K. variicola (n = 33), K. aerogenes (n = 1), Raoultella ornithinolytica (n = 38) and R. planticola (n = 13) were isolated. The number of positive samples increased with higher levels of faecal contamination. We found low prevalence of acquired resistance in all isolates, with seven K. pneumoniae isolates showing resistance to more than one antibiotic class. The complete genome sequence of cefotaxime-resistant K. pneumoniae sensu stricto isolate 2016-1400 was obtained using Oxford Nanopore and Illumina MiSeq based sequencing. The 2016-1400 genome had two contigs, one chromosome of 5,088,943 bp and one plasmid of 191,744 bp and belonged to ST1035. The β-lactamase genes blaCTX-M-3 and blaTEM-1, as well as the heavy metal resistance genes pco, ars and sil were carried on a plasmid highly similar to one found in K. pneumoniae strain C17KP0055 from South-Korea recovered from a blood stream infection. The present study demonstrates that K. pneumoniae are prevalent in the coastal marine environment and that bivalve molluscs may act as a potential reservoir of extended spectrum β-lactamase (ESBL)-producing K. pneumoniae that may be transmitted through the food chain. [ABSTRACT FROM AUTHOR]

In 2015, tetrodotoxins (TTXs) were considered a potential threat in Europe since several studies had shown the presence of these toxins in European bivalve molluscs. In this study, we investigated the occurrence of TTXs in 127 bivalve samples (mussels and oysters) and in 66 gastropod samples (whelks) collected all along the French mainland coasts in 2017 and 2018. Analyses were carried out after optimization and in-house validation of a performing hydrophilic interaction liquid chromatography associated with tandem mass spectrometry (HILIC-MS/MS) method. The concentration set by European Food Safety Authority (EFSA) not expected to result in adverse effects (44 µg TTX equivalent/kg) was never exceeded, but TTX was detected in three mussel samples and one whelk sample (1.7–11.2 µg/kg). The tissue distribution of TTX in this whelk sample showed higher concentrations in the digestive gland, stomach and gonads (7.4 µg TTX/kg) than in the rest of the whelk tissues (below the limit of detection of 1.7 µg TTX/kg). This is the first study to report the detection of TTX in French molluscs. [ABSTRACT FROM AUTHOR]

Abstract: The aim of this study was to (a) evaluate the potential for the ‘Comet assay’ to be used as a method for detecting genetic damage in the common cockle Cerastoderma edule; and (b) to compare the relative sensitivity with Mytilus edulis as the bivalve widely used as a sentinel species in biomonitoring studies. In vitro validation studies were carried out on haemocytes from each species using hydrogen peroxide (H2O2), a known oxidant and the induced DNA damage was measured using the Comet assay. On exposure to 0, 100, 500 and 1000μM H2O2, a significant concentration-dependent increase was observed in both species. Use of an additional concentration of 5000μM H2O2 showed that while DNA damage could be assessed in M. edulis at this concentration, only a few cells from C. edule were amenable to measurements owing to extensive DNA damage (“hedgehog cells”). The evidence also suggested that the cells from C. edule are more sensitive to oxidative damage induced by H2O2 when compared with M. edulis. Bearing in mind that sediments are the ultimate sink for many contaminants, this study demonstrates the potential application of sediment-dwelling C. edule as a useful biomonitoring species. [Copyright &y& Elsevier]