Aequipecten Opercularis Research Articles

Lactiplantibacillus plantarum I Induces Gonad Growth in the Queen Scallop Aequipecten opercularis (Linnaeus, 1758) under Conditions of Climate Change

Climate change has presented a serious problem in recent times, which is why a new approach is being sought in terms of aquacultural food quality. In this study, the influence of temperature increase (by 2 °C) and pH decrease (by 0.2) was investigated on the queen scallop, Aequipecten opercularis (Linnaeus, 1758). Furthermore, the effect of a food-enriched diet with the probiotic culture Lactiplantibacillus plantarum I was assessed in climate-changed conditions. Scallops’ morphometric parameters were measured before the experimental setup and after one month of being kept in controlled conditions. Morphometric parameters included the elongation index, compactness index, convexity index, density index, condition index, meat yield, gonadosomatic index, adductor muscle index, and hepatosomatic index. Climate-changed conditions had no effect on the scallop condition index, meat yield, or hepatosomatic index. Nevertheless, the addition of probiotics to their diet had a positive effect on the queen scallops cultivated under conditions of climate change, influencing positive allometry and the increase of the gonadosomatic indices. On the other hand, the same conditions negatively affected the adductor muscle index of the scallops. To conclude, in the context of climate change conditions, queen scallops could be a good organism of choice that can be very well adapted to the changed environmental conditions, especially with the addition of the lactic acid bacteria culture Lpb. plantarum I.

The queen scallop Aequipecten opercularis: A slow domoic acid depurator?

Domoic acid (DA) is a dangerous phycotoxin produced by several strains of diatoms of the genus Pseudo-nitzschia, and responsible for Amnesic Shellfish Poisoning (ASP) in humans. The increasingly intense ASP-outbreaks along the English Channel over the last three decades have forced persistent harvest closures of economically important and highly contaminated bivalve stocks exhibiting slow DA-depuration rates, like the king scallop Pecten maximus. Under this scenario, other pectinid species, such as the queen scallop Aequipecten opercularis have been empirically proposed as alternative resources to redress the high economic losses due to the banning of the exploitation of P. maximus. Nevertheless, the kinetics of DA depuration in A. opercularis have not been assessed so far, and its direct extraction after ASP-episodes could represent a serious threat to public health. Hence, the main objective of this work was to estimate the DA-depuration rate in the digestive gland (DG) of naturally contaminated scallops A. opercularis after a toxic Pseudo-nitzschia australis bloom subjected to experimental depuration in the laboratory for 30 days. This study also intended to go further in the knowledge about the anatomical distribution of DA in scallop tissues, and corroborate the implications of autophagy in DA-sequestration in the DG of this species as recently hypothesized. In the DG, the DA-depuration rate (0.018 day−1) suggested that even with toxin burdens as low as 40 mg⋅kg−1 in the DG, queen scallops may remain contaminated for about 70 days, thus longer under intensely contamination scenarios. The subcellular analyses corroborated DA-sequestration mainly through late-autophagy within residual bodies in the DG, without differences in the frequencies of anti-DA labeled residual bodies across the entire depuration process. These results revealed that A. opercularis cannot be considered a fast DA-depurator, and represent a baseline knowledge for decision-making about harvesting natural beds of queen scallops after toxic Pseudo-nitzschia blooms. The findings of this work also represent a cornerstone for further research to accelerate DA-depuration in this species.

Procjena utjecaja matriksa na kvantifikaciju lipofilnih toksina u plodovima mora

Phycotoxin accumulation in seafood can cause human intoxication and significant economic losses in seafood-producing areas. To protect consumer safety, maximum permitted levels in bivalve molluscs have been set, with appropriate analytical methods for their determination and quantification. The reference method for lipophilic phycotoxins, commonly referred to as lipophilic toxins, is liquid chromatography tandem mass spectrometry (LCMS/MS method), where coeluting components from the matrix can affect the efficiency of ionisation, resulting in erroneous quantification. In this study, the matrix effect was evaluated using the slope ratio analysis method, which involves comparing the slopes of the calibration curves in the matrix with those in pure solvent. The extent of the matrix effect was investigated in mussels, oysters, queen scallops and ascidians. The study covered all phycotoxins under Regulation EU 853/2004 for which certified standards are commercially available, including unregulated pectenotoxin 2. The results indicated that all lipophilic toxins were susceptible to this effect when LC-MS/MS was used for their determination. Significant ion suppression was evident for most analytes in all matrices, except for okadaic acid and dinophysistoxin 2 in bivalves, where significant ion enhancement was demonstrated, and dinophysistoxin 1 in oyster and scallop extract where no significant effect on ionisation was observed. Further analysis revealed no significant differences between the slope of mussel matrix-matched calibration and that of other bivalve matrices. Given this minor difference, the mussel matrix-matched calibration curve could be applied to minimise the matrix effect and to quantify phycotoxins in bivalve matrices analysed here, with the exception of the okadaic acid group in ascidians, which requires matrix-matched calibration prepared with the blank extract of these mentioned species. Given the risks phycotoxins pose to human health, ongoing analytical method development is necessary in this field to properly control food safety and ensure consumer health.

Projecting expected growth period of bivalves in a coastal temperate sea

AbstractThe impact of climate warming on coastal benthic fauna is already observed, but forecasting their long‐term fate remains challenging. This study uses δ18Oshell data of specimens of five bivalve species collected at six locations and results from kilometer‐scale atmosphere–ocean climate model for the time intervals of 1987–2017 and 2070–2100, to estimate changes in bivalve growth phenology. All species will benefit from climate warming during winter, experiencing a longer growing season than currently. The growth of Aequipecten opercularis, Flexopecten glaber, and Pecten jacobaeus will decrease in summer, resulting in up to 3 months of reduced growth per year. Glycymeris pilosa and Venus verrucosa in the southern Adriatic Sea will be more affected than those in the north, with up to 4 months longer annual growth. These findings can inform adaptation plans for bivalve management in the Adriatic Sea but also in areas where the studied species are present.

Effect of Dietary Supplementation with Lactiplantibacillus plantarum I On Queen Scallop Aequipecten opercularis Under Simulated Climate Change Conditions

Abstract This study examined the effects of dietary supplementation of queen scallop Aequipecten opercularis with an indigenous strain of lactic acid bacteria (LAB), Lactiplantibacillus plantarum I, previously isolated from its digestive tract, on gut microbial populations and growth rates during cultivation under simulated climate change conditions (pH 7.8, T = 16 ± 2 °C). After one month of feeding, the results showed a noticeable reduction in aquaculture diseases causing pathogens while maintaining sufficient viable Lpb. plantarum I cells. A higher pH and temperature resulted in higher growth rates, measured by the weight and length of scallops, compared to the control group. The results obtained shed light on the influence of the addition of lactic acid bacteria on the growth of bivalve mollusks under normal and climate change conditions, and provide control of pathogenic microorganisms. In the context of climate change, host-pathogen interactions need to be recognized and put under control by applying natural solutions to minimize the environmental footprint.

Comparative study of domoic acid accumulation, isomer content and associated digestive subcellular processes in five marine invertebrate species

Despite the deleterious effects of the phycotoxin domoic acid (DA) on human health, and the permanent threat of blooms of the toxic Pseudo-nitzschia sp. over commercially important fishery-resources, knowledge regarding the physiological mechanisms behind the profound differences in accumulation and depuration of this toxin in contaminated invertebrates remain very scarce. In this work, a comparative analysis of accumulation, isomer content, and subcellular localization of DA in different invertebrate species was performed. Samples of scallops Pecten maximus and Aequipecten opercularis, clams Donax trunculus, slippersnails Crepidula fornicata, and seasquirts Asterocarpa sp. were collected after blooms of the same concentration of toxic Pseudo-nitzschia australis. Differences (P < 0.05) in DA accumulation were found, wherein P. maximus showed up to 20-fold more DA in the digestive gland than the other species. Similar profiles of DA isomers were found between P. maximus and A. opercularis, whereas C. fornicata was the species with the highest biotransformation rate (∼10 %) and D. trunculus the lowest (∼4 %). DA localization by immunohistochemical analysis revealed differences (P < 0.05) between species: in P. maximus, DA was detected mainly within autophagosome-like vesicles in the cytoplasm of digestive cells, while in A. opercularis and C. fornicata significant DA immunoreactivity was found in post-autophagy residual bodies. A slight DA staining was found free within the cytoplasm of the digestive cells of D. trunculus and Asterocarpa sp. The Principal Component Analysis revealed similarities between pectinids, and a clear distinction of the rest of the species based on their capabilities to accumulate, biotransform, and distribute the toxin within their tissues. These findings contribute to improve the understanding of the inter-specific differences concerning the contamination-decontamination kinetics and the fate of DA in invertebrate species.

The Effect of Lactiplantibacillus plantarum I-Enriched Diet on the Phenolic Content and Antioxidant Capacity of Queen Scallop (Aequipecten opercularis Linnaeus, 1758) Extracts.

The use of probiotics in the diet of bivalves poses a great potential in aquaculture as an alternative to antibiotics. The aim of this study was to assess the effect of Lactiplantibacillus plantarum I on the phenolic content and antioxidant capacity (AC) of queen scallop extracts after one month of feeding. Total phenols (TP) ranged from 28.17 ± 3.11 to 58.58 ± 8.57 mg GAE/100 g, total non-flavonoids (TNF) from 23.33 ± 3.66 to 36.56 ± 9.91 mg GAE/100 g, and total flavonoids (TF) from 10.56 ± 5.57 to 30.16 ± 1.69 mg CE/100 g. AC was assessed via three different methods: the ferric-reducing ability of plasma assay (FRAP), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic) acid assay (ABTS), and 2,2-diphenyl-1-picryhydrazyl assay (DPPH). FRAP values ranged from 0.13 ± 0.03 to 0.17 ± 0.02 µM AA/g, ABTS from 0.68 ± 0.11 to 2.79 ± 0.34 µM AA/g, and DPPH from 1.75 ± 0.17 to 2.98 ± 0.53 µM AA/g. Among all extracts, the best phenolic content and AC were observed in water extracts from queen scallops. The bivalves treated with the Lactiplantibacillus plantarum I-enriched diet showed higher AC according to the FRAP assay in all extracts. A significant correlation was observed between AC and TP and TNF in control and Lactiplantibacillus plantarum I-treated scallops.

Study of the Impact of Lactiplantibacillus plantarum I on the Health Status of Queen Scallop Aequipecten opercularis

The objective of this research was to isolate and identify lactic acid bacteria living in the digestive tract of the queen scallop Aequipecten opercularis and further test it as a probiotic culture on the growth of the bivalve during one month in captivity. Classic microbiological techniques (API and MALDI TOF) were used and four different strains of Lactiplantibacillus and one Lactococcus were identified and further analyzed, namely: Lactiplantibacillus plantarum I, Lactiplantibacillus plantarum 1, Lactiplantibacillus plantarum 2, Lactococcus lactis, and Lactiplantibacillus brevis. The isolated cultures were further tested for the highest antimicrobial activity towards the most common marine pathogens and survival at different temperatures and pH levels. The strain Lactiplantibacillus plantarum I had the best results, with the highest antimicrobial activity (77–85% pathogen growth inhibition) and the best safety standards showing no antibiotic resistance, and no ability to synthesize biogenic amine and degrade red blood cells. Considering all the abovementioned characteristics, the strain Lpb. plantarum I was further tested on the growth and health status of the bivalve Aequipecten opercularis during a month of cultivation in captivity. The results showed that the incorporation of Lpb. plantarum I led to a decrease in the quantity of test microorganisms in the bivalve and an increase in both the growth rate weight and growth rate length of the queen scallop A. opercularis. Furthermore, the scallops fed with the algae culture + Lpb. plantarum I gained significantly higher meat yield (33.15 ± 2.63%) compared with the control scallops fed only with the algae culture (29.66 ± 2.87%). To conclude, the results from this research indicate that including Lpb. plantarum I as a dietary supplement could enhance growth performance and serve as a feasible approach to reduce pathogen levels while cultivating A. opercularis in captivity.

Distribution of metals in the queen scallop Aequipecten opercularis during a transplant experiment: Metal rich granules as drivers of Pb bioaccumulation

The queen scallop Aequipecten opercularis accumulates high concentrations of lead (Pb) in its tissues, what has led to the interruption of this fishery in some extraction areas in Galicia (NW Spain). This study follows the dynamics of bioaccumulation of Pb and other metals in this species, the tissue distribution and the subcellular partitioning in selected organs, in order to understand the mechanisms that provoke the high Pb levels reached in its tissues and to increase our knowledge about metal bioaccumulation dynamics in this species. Scallops originating from a clean area were exposed in cages in two places in the Ría de Vigo (one shipyard and a less impacted location) and 10 individuals were collected every month over a three months period. Metal bioaccumulation and metal distribution in several organs, including gills, digestive gland, kidneys, muscle, gonad and remaining tissues, was studied. The results showed that scallops accumulated similar levels of Cd, Pb and Zn at both sites, while Cu and Ni showed an opposite pattern at the shipyard, with Cu concentrations increasing around 10 times and Ni decreasing during the 3 months of exposure. The preferential organs for metal accumulation were the kidneys for Pb and Zn, the digestive gland for Cd, both organs for Cu and Ni, and the muscle for As. Subcellular partitioning of kidney samples additionally showed an extraordinary ability to accumulate Pb and Zn at very high concentrations in kidney granules, a fraction that accounted for 30 to 60 % of Pb in soft-tissues. It is concluded that Pb bioaccumulation in kidney granules is the mechanism responsible for the high levels of Pb observed in this species.

Commercial Quality, Biological Indices and Biochemical Composition of Queen Scallop Aequipecten opercularis in Culture

Market demand for scallops has considerably increased in recent decades. Although natural populations of scallops are vulnerable, the queen scallop, Aequipecten opercularis, can represent a possible alternative to at-risk species. The aim of this study was to determine the effect of seawater parameters on the commercial quality, biological indices and nutritional quality of the scallop A. opercularis in 1900 L tanks in ex situ conditions. The condition index (CI) and meat yield (MY) peaked in autumn (89.92% and 40.29%, respectively). The muscle index (MI) showed the highest peak during the winter season (5.96%), while the gonadosomatic index (GSI) (34.06%) peaked in the spring months. Protein content (6.89–9.56 g/100 g), lipids (2.58–2.79 g/100 g) and carbohydrates (0.12–0.33 g/100 g) varied during the study period. Seasonal fluctuations in seawater temperature and pH negatively influenced the CI, MY and protein and lipid content, while positively affecting carbohydrate and moisture content. The most suitable period of the year for the consumption of the scallop A. opercularis was found to be the winter and spring periods, when the scallops achieved their highest nutritional value. It can be stated from this study that queen scallops in the Adriatic Sea have a high nutritional quality and that they can be proposed for higher human consumption.

THE FIRST RECORD OF THE SCALLOP, &lt;i&gt;AEQUIPECTEN OPERCULARIS&lt;/i&gt; (L. 1758) (MOLLUSCA, PECTINIDAE), IN RUSSIA’S WATERS

A juvenile specimen of the scallop, Aequipecten opercularis, was found in 2007 in the eastern part of the Varanger-Fjord, Kola coast of the Barents Sea. The northern range limit of this species has earlier been drawn much more southerly, at the Atlantic coast of Norway. This record is the first in Russia’s waters, probably being related to climate warming in the southern part of the Barents Sea since the early 2000’s.

First Data on Campylobacter spp. Presence in Shellfish in Croatia.

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.

Effects of material composition and face block exposure on the long-term (2014-2018) colonisation of an intertidal RECIF Artificial Reef in the Bay of Seine

An experimental artificial reef (AR) with the incorporation of crushed seashells of the queen scallop Aequipecten opercularis was initiated on the intertidal zone of the Bay of Seine in the framework of the INTERREG IVa RECIF project for a everyone year monitoring observations from March 2014 to April 2018. Three block types were operated: eco-friendly material with 20% or 40% of crushed queen scallop shells, and ordinary concrete made from natural aggregates. On the seven blocks analysed at the end of the experiment in 2018, 74 taxa including 32 sessile and 42 motile fauna have been accounted; no difference of taxonomic richness was observed between material and immersion time. The sessile fauna was identified for the six external faces of two blocks aged of four years: horizontal above face (FAB), horizontal below Face (FBE), face oriented offshore (FOF), face oriented inshore (FIN), vertical face oriented west (FEW) and vertical face oriented east (FEA). FBE showed higher taxonomic richness and abundance than the other block faces. FAB exposed to the light showed the lowest abundances. Our observations showed the efficiency of such AR on the intertidal zone accessible at each spring tide, which was easier to survey than in the subtidal zone, where divers must be mobilised to sample blocks.

Valorization of queen scallop shells in the preparation of metakaolin-based geopolymer mortars

This paper aims to assess the effect of the seashell waste (SW) incorporation on the performance of MK-based geopolymer mortar. The SW powder was introduced as partial replacement of MK with a substitution rate of 0, 10, 20 and 30%. Its effect on setting time, water porosity, mechanical strengths and both drying and autogenous shrinkage was investigated. A special attention was also devoted to the evaluation of the efflorescence stability. The obtained results show that partial replacement of MK by SW powder leads to limit considerably the water demand of the binder resulting in mortars with higher mechanical strengths and lower porosity and shrinkage. In addition, SW powder allows to accelerate the setting time of geopolymer mortars. The autogenous shrinkage was found to increase with increasing SW percentage. However, the total shrinkage was dominated by the drying shrinkage that represents around 85% of the total strain at 90 days regardless the SW replacement rate. In terms of efflorescence stability, the lowest efflorescence formation was attributed to the geopolymer mixture formulated with 30% of SW powder.

Settlement of Bivalve Spat on Artificial Collectors (Net Bags) in Two Commercial Mussel Parks in the North-Western Adriatic Sea

Among aquaculture activities, shellfish culture is considered more sustainable and beneficial in terms of food security. Currently, only a few bivalve species are reared and there is a need to explore the possibility to introduce new candidates for shellfish farming. Due to the lack of information on bivalve recruitment in the North-Western Adriatic Sea, in this study, the possibility to collect natural spat of commercial species was investigated. Artificial collectors (net bags) were deployed in two sites, Pellestrina and Caleri (North-Western Adriatic Sea), within two commercial mussel parks, during the spring–summer and summer–autumn periods. At both sites, collectors were placed at a distance of 1 m from each other, from 5 to 14 m depth. The influence of season, site and depth on bivalve recruitment was inspected and the presence of invasive species was also evaluated. In all, 28 bivalve taxa were found, and a higher settlement rate was observed in summer–autumn compared to the spring–summer period. Mytilus galloprovincialis, Flexopecten glaber, Mimachlays varia and Aequipecten opercularis were the most abundant species in spring–summer. In the summer–autumn period, in both sites analysed, a very high quantity of Anadara transversa and F. glaber were found. Indeed, these species were dominant at Pellestrina and Caleri, respectively. Another non-indigenous species, Arcuatula senhousia, was also detected. Relevant amounts of Pectinidae spat, F. glaber in particular, were collected and the optimal depth range for the scallop spat collection was found to be between 8 and 14 m. Our results highlight the relevant potential of Pectinidae spat collection along the North-Western Adriatic coasts, even though the presence of invasive species needs to be monitored.

Larvae of Sulcascaris sulcata (Nematoda: Anisakidae), a parasite of sea turtles, infect the edible purple dye murex Bolinus brandaris in the Tyrrhenian Sea

Anisakid nematodes are among the most common helminth parasites of the marine realm, particularly important for their implications with human infections and/or allergies. Members of the Anisakidae use aquatic mammals, birds and fish as definitive hosts, and crustaceans, fish and molluscs as intermediate/paratenic hosts. Sulcascaris sulcata, the only species in the monotypic genus Sulcascaris, represents the exception being a parasite of sea turtles as adult. The recent findings of larvae of S. sulcata in scallops (Pecten jacobeus and Aequipecten opercularis) and Mediterranean mussels Mytilus galloprovincialis intended for human consumption from the Mediterranean Sea caused concern regarding the sanitary control of edible molluscs and consumer safety. Herein, we investigated the larval anisakids collected from the purple dye murex, Bolinus brandaris, harvested for human consumption from the Central Mediterranean Sea (Tyrrhenian Sea). Morphological study and sequence analysis of the internal transcribed spacer regions of the ribosomal DNA and the mitochondrial cytochrome c oxidase subunit 2 gene locus revealed the occurrence of fourth stage larvae of S. sulcata in 16% of the purple dye murex examined. The present study adds the purple dye murex to the list of the known intermediate hosts of this parasite in the Mediterranean Sea and the northern coast of Campania region as site where individuals of loggerhead turtle and purple dye murex may become infected. This is the first study reporting an anisakid nematode in edible gastropods. Epidemiological features of infection in the purple dye murex and implications for gastropod safety and risk for consumers are discussed.

Occurrence and Seasonal Monitoring of Domoic Acid in Three Shellfish Species from the Northern Adriatic Sea.

As filter feeders, bivalves and ascidians can accumulate contaminants present in the environment and pass them on to higher food chain levels as vectors. The consumption of bivalves contaminated with the potent neurotoxin domoic acid (DA) can cause amnesic shellfish poisoning in humans. The aim of this study was to determine seasonal differences in occurrence and accumulation of this phycotoxin in European oysters (Ostrea edulis Linnaeus, 1758) (n = 46), Queen scallops (Aequipecten opercularis Linnaeus, 1758) (n = 53), and edible ascidians of the Microcosmus spp. (n = 107), originating from the same harvesting area in the Northern Adriatic Sea. The quantification was performed using ultra-performance liquid chromatography–tandem mass spectrometry (LC-MS/MS) preceded by derivatization with dansyl chloride. DA was found in very low concentrations throughout the year, with a maximum value of 810 μg/kg in Queen scallops. This study reveals differences in the occurrence and accumulation of DA between Queen scallops and the other two investigated species (oysters and ascidians) and the highest concentrations during the colder part of the year. Even though DA was detected in all of them, Queen scallops showed higher DA accumulation compared to the other two (p < 0.001), hence representing a sentinel species suitable for the monitoring of DA level in seafood.

Quantifying abundance trends and environmental effects on a population of queen scallop Aequipecten opercularis targeted by artisanal fishers in a coastal upwelling area (Ría de Arousa, NW Spain) using a Bayesian spatial model

Queen scallop Aequipecten opercularis, is exploited by small-scale trawlers in separated aggregations along the east Atlantic coast and the Mediterranean Sea. However, population performance is poorly known. Here, we combine official information and on-board observers’ data over two decades to study the fishery ecology of an aggregation occurring in a coastal upwelling system (Ría de Arousa, NW Spain). Annual landings fluctuated around a mean of 170 tons while beam trawlers declined at a rate of 11 vessels per decade. A hurdle Bayesian spatial model fitted to observers’ catch and effort data showed that the probability of occurrence and abundance of the species increased with fishing effort and decreased in sandy bottoms. Moreover, abundance increased with upwelling intensity and decreased with continental runoff and along the fishing season. Furthermore, occurrence was higher in the inner part of the embayment while abundance increased in the central channel. The predicted index of standardized abundance correlated with the trend in landings, and year-to-year fluctuations in abundance were negatively and positively related to upwelling intensity and net primary production, respectively, during the spawning and settlement period.

Evaluation of the shucking of certain species of scallops contaminated with lipophilic toxins with a view to the production of edible parts meeting the safety requirements foreseen in the Union legislation.

EFSA was asked by the European Commission to provide information on levels of lipophilic shellfish toxins in whole scallops that would ensure levels in edible parts below the regulatory limits after shucking, i.e. removal of non‐edible parts. This should include the okadaic acid (OA), the azaspiracid (AZA) and the yessotoxin (YTX) groups, and five species of scallops. In addition, EFSA was asked to recommend the number of scallops in an analytical sample. To address these questions, EFSA received suitable data on the three toxin groups in two scallop species, Aequipecten opercularis and Pecten maximus, i.e. data on individual and pooled samples of edible and non‐edible parts from contamination incidents. The majority of the concentration levels were below limit of quantification (LOQ)/limit of detection (LOD), especially in adductor muscle but also in gonads. Shucking in most cases resulted in a strong decrease in the toxin levels. For Pecten maximus, statistical analysis showed that levels in whole scallops should not exceed 256 μg OA eq/kg or 217 μg AZA1 eq/kg to ensure that levels in gonads are below the regulatory limits of 160 μg OA or AZA1 eq/kg with 99% certainty. Such an analysis was not possible for yessotoxins or any toxin in Aequipecten opercularis and an assessment could only be based on upper bound levels. To ensure a 95% correct prediction on whether the level in scallops in an area or lot is correctly predicted to be compliant/non‐compliant, it was shown that 10 scallops per sample would be sufficient to predict with 95% certainty if levels of OA‐group toxins in the area/lot were 25% below or above the regulatory limit. However, to predict with a 95% certainty for levels between 140 and 180 μg OA eq/kg, a pooled sample of more than 30 scallops would have to be tested.

Cyclic imines in shellfish and ascidians in the northern Adriatic Sea

ABSTRACT The aim of this study was to determine the occurrence of the most representative cyclic imines (CIs) gymnodimine (GYM), pinnatoxin G (PnTX-G), and 13-desmethyl SPX C (SPX1) in Mediterranean mussels (Mytilus galloprovincialis Lamarck, 1819) (n = 416), European oysters (Ostrea edulis Linnaeus, 1758) (n = 104), Queen scallops (Aequipecten opercularis Linnaeus, 1758) (n = 52) and edible ascidians of the Microcosmus spp. (n = 104) originating from nine harvesting and breeding areas in the northern part of the Adriatic Sea using ultra-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). All CI concentrations were far below the guidance level of 400 μg SPXs/kg proposed by the EU Reference Laboratory for Marine Toxins. In contrast to Queen scallops and ascidians, in Mediterranean mussels and European oysters CIs were found throughout the year. Our data reveal the differences between species predisposed for CIs accumulation, as well as seasonal and locational variations in CIs occurrence.