Anthropogenic pollutants in Nephrops norvegicus (Linnaeus, 1758) from the NW Mediterranean Sea: Uptake assessment and potential impact on health.

Anthropogenic pollution is considered one of the main threats to the marine environment, and there is an imperious need to assess its potential impact on ecologically and economically relevant species. This study characterises plastic ingestion and tissue levels of potentially toxic metallic elements in Nephrops norvegicus and their simultaneous levels in abiotic compartments from three locations of the Catalan coast (NW Mediterranean Sea). A multidisciplinary assessment of the health condition of N. norvegicus through condition indices, enzymatic biomarkers and histological techniques is provided, and its relationship with anthropogenic pollutant levels explored. Plastic fibres were commonly found in stomachs of N. norvegicus (85% of the individuals), with higher abundances (13 ± 21 fibres · ind-1) in specimens captured close to Barcelona. The presence of long synthetic fibres in near-bottom waters, as well as the mirroring trends in abundance among locations for water and ingested plastics, suggest that uptake from water may be occurring potentially through suspension feeding. The spatial variability in the levels of metallic elements in N. norvegicus was poorly correlated to the variability in sediments. In any case, present levels in abdominal muscle are considered safe for human consumption. Levels of ingested plastics only showed significant, yet weak, correlations with glutathione S-transferase and catalase activities. However, no other health parameter analysed showed any trend potentially associated to anthropogenic pollutant levels. Neither the condition indices nor the histopathological assessment evidenced any signs of pathologic conditions affecting N. norvegicus. Thus, it was concluded that presently there is no evidence of a negative impact of the studied pollutants on the health condition of N. norvegicus in the studied grounds.

A Novel Detection Refinement Technique for Accurate Identification of Nephrops norvegicus Burrows in Underwater Imagery.

With the evolution of the convolutional neural network (CNN), object detection in the underwater environment has gained a lot of attention. However, due to the complex nature of the underwater environment, generic CNN-based object detectors still face challenges in underwater object detection. These challenges include image blurring, texture distortion, color shift, and scale variation, which result in low precision and recall rates. To tackle this challenge, we propose a detection refinement algorithm based on spatial-temporal analysis to improve the performance of generic detectors by suppressing the false positives and recovering the missed detections in underwater videos. In the proposed work, we use state-of-the-art deep neural networks such as Inception, ResNet50, and ResNet101 to automatically classify and detect the Norway lobster Nephrops norvegicus burrows from underwater videos. Nephrops is one of the most important commercial species in Northeast Atlantic waters, and it lives in burrow systems that it builds itself on muddy bottoms. To evaluate the performance of proposed framework, we collected the data from the Gulf of Cadiz. From experiment results, we demonstrate that the proposed framework effectively suppresses false positives and recovers missed detections obtained from generic detectors. The mean average precision (mAP) gained a 10% increase with the proposed refinement technique.

Size dependent egestion of polyester fibres in the Dublin Bay Prawn (Nephrops norvegicus).

Microplastics (MPs) are an extensive global contaminant in the marine environment, known to be ingested by marine organisms. The presence of MPs in the commercially important marine decapod crustacean Nephrops norvegicus (Dublin Bay Prawn) has been documented for the North-East Atlantic and the Mediterranean, however, uncertainties remain about retention times of MPs in the gastrointestinal tract (GIT) of this species. This study aims to investigate the retention times of polyester MP fibres of three sizes (3, 5, and 10 mm in length) and to determine whether the egestion of MP fibres is size and time dependent. Results suggest that MP fibres of different lengths are retained for different periods of time, with larger MP fibres being retained for longer periods (e.g., minimum 96 h for 10 mm fibres). The present study also assesses for the first time, the size dependent relationship of MP fibres under controlled conditions for N. norvegicus.

A baseline study of the metallothioneins content in digestive gland of the Norway lobster Nephrops norvegicus from Northern Adriatic Sea: Body size, season, gender and metal specific variability.

Metallothioneins content was investigated in digestive gland of two wild-caught Norway lobster Nephrops norvegicus populations from the Northern Adriatic Sea, in relation to body size, season and gender. Concomitant accumulation of cadmium, mercury, arsenic, lead, chromium and manganese, reactive oxygen species concentration and energy reserves in digestive gland were also assessed. While differences between genders were not recorded, metallothioneins content seasonal trends were affected by body size. Most of parameters displayed inconsistent trends across sampling sites. Significant correlation between metallothioneins content and cadmium, arsenic and mercury concentrations was recorded only for larger lobsters. A negative correlation of reactive oxygen species concentration and metallothioneins content was observed for small, but not large lobsters. Energy reserves, in particular lipids, could considerably influence biochemical and chemical parameters variations. The present results constitute the essential baseline for future studies aimed at evaluating the N. norvegicus health in relation to metal contamination of coastal sediments.

Bioaccumulation of PCBs from microplastics in Norway lobster (Nephrops norvegicus): An experimental study.

Plastic debris acts as a sorbent phase for hydrophobic organic compounds like polychlorinated biphenyls (PCBs). Chemical partitioning models predict that the ingestion of microplastics with adsorbed chemicals in the field will tend not to result in significant net desorption of the chemical to the organism's tissues. This is expected due to the often limited differences in fugacity of the chemical between the indigestible plastic materials and the tissues, which are typically already exposed in the same environment to the same chemicals as the plastic. However laboratory trials validating these model predictions are scarce. In this study, PCB-loaded microplastics were offered to field-collected Norway lobsters (Nephrops norvegicus) during in vivo feeding laboratory experiments. Each ingestion experiment was repeated with and without loading a mixture of ten PCB congeners onto plastic microspheres (MS) made of polyethylene (PE) and polystyrene (PS) with diameters of either 500-600 µm or 6 µm. We observed that the presence of chemicals adsorbed to ingested microplastics did not lead to significant bioaccumulation of the chemicals in the exposed organisms. There was a limited uptake of PCBs in Nephrops tail tissue after ingestion of PCB-loaded PE MS, while almost no PCBs were detected in animals exposed to PS MS. In general, our results demonstrated that after 3 weeks of exposure the ingestion of plastic MS themselves did not affect the nutritional state of wild Nephrops.

Sex and tissue specific gene expression patterns identified following de novo transcriptomic analysis of the Norway lobster, Nephrops norvegicus.

BACKGROUND: The Norway lobster, Nephrops norvegicus, is economically important in European fisheries and is a key organism in local marine ecosystems. Despite multi-faceted scientific interest in this species, our current knowledge of genetic resources in this species remains very limited. Here, we generated a reference de novo transcriptome for N. norvegicus from multiple tissues in both sexes. Bioinformatic analyses were conducted to detect transcripts that were expressed exclusively in either males or females. Patterns were validated via RT-PCR. RESULTS: Sixteen N. norvegicus libraries were sequenced from immature and mature ovary, testis and vas deferens (including the masculinizing androgenic gland). In addition, eyestalk, brain, thoracic ganglia and hepatopancreas tissues were screened in males and both immature and mature females. RNA-Sequencing resulted in >600 million reads. De novo assembly that combined the current dataset with two previously published libraries from eyestalk tissue, yielded a reference transcriptome of 333,225 transcripts with an average size of 708 base pairs (bp), with an N50 of 1272 bp. Sex-specific transcripts were detected primarily in gonads followed by hepatopancreas, brain, thoracic ganglia, and eyestalk, respectively. Candidate transcripts that were expressed exclusively either in males or females were highlighted and the 10 most abundant ones were validated via RT-PCR. Among the most highly expressed genes were Serine threonine protein kinase in testis and Vitellogenin in female hepatopancreas. These results align closely with gene annotation results. Moreover, a differential expression heatmap showed that the majority of differentially expressed transcripts were identified in gonad and eyestalk tissues. Results indicate that sex-specific gene expression patterns in Norway lobster are controlled by differences in gene regulation pattern between males and females in somatic tissues. CONCLUSIONS: The current study presents the first multi-tissue reference transcriptome for the Norway lobster that can be applied to future biological, wild restocking and fisheries studies. Sex-specific markers were mainly expressed in males implying that males may experience stronger selection than females. It is apparent that differential expression is due to sex-specific gene regulatory pathways that are present in somatic tissues and not from effects of genes located on heterogametic sex chromosomes. The N. norvegicus data provide a foundation for future gene-based reproductive studies.

Lipid Composition of Oil Extracted from Wasted Norway Lobster (Nephrops norvegicus) Heads and Comparison with Oil Extracted from Antarctic Krill (Euphasia superba).

In the UK, the Norway lobster (Nephrops norvegicus) supports its most important shellfish fishery. Nephrops are sold either whole, or as "tails-only" for the scampi trade. In the "tailing" process, the "head" (cephalothorax) is discarded as waste. A smaller crustacean species, the Antarctic krill Euphasia superba, represents an economically valuable industry, as its extractable oil is sold as a human dietary supplement. The aim of this study was to determine the amount and composition of the oil contained in discarded Nephrops heads and to compare its composition to the oil extracted from krill. Differences due to Geographical variation and seasonal patterns in the amount and composition of lipid were also noted. Results indicated that Nephrops head waste samples collected from more southern locations in Scotland (Clyde Sea area) contained higher levels of oil when compared to samples collected from northern locations in Iceland. Moreover, seasonal differences within the Clyde Sea area in Scotland were also observed, with oil extracted from Nephrops head waste peaking at around 11.5% during the summer months when larger and more mature females were caught by trawl. At this time of the year, the valuable fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) accounted for around 23% of the total fatty acid content in oil extracted from Nephrops head waste. A seasonal effect on EPA content was found, with higher levels obtained in the summer, while no trend was found in DHA percentages. Finally, oil from Nephrops head waste contained a higher proportion of EPA and DHA than krill oil but these fatty acids were more abundantly linked to the neutral lipids rather to than polar lipids. The characterization of lipid that could be extracted from Nephrops head waste should be seen as a first step for the commercial use of a valuable resource currently wasted. This approach is extremely relevant given the current limited supply of EPA and DHA and changes in the Common Fisheries Policy.

Long-term microplastic retention causes reduced body condition in the langoustine, Nephrops norvegicus.

Microplastic represents a rising proportion of marine litter and is widely distributed throughout a range of marine habitats. Correspondingly, the number of reports of species containing microplastics increases annually. Nephrops norvegicus in the Firth of Clyde have previously been shown to retain large aggregations of microplastic fibres. The potential for N. norvegicus to retain plastic over an extended time period increases the likelihood of any associated negative impacts to the individual. This study represents the longest observation of the impacts of microplastic retention in invertebrates. We exposed N. norvegicus to plastic over eight months to determine the impacts of extended exposure. Over this period we compared the feeding rate, body mass, and nutritional state of plastic-fed N. norvegicus to that of fed and starved control groups. Following the experimental period, the plastic-fed langoustine contained microplastic aggregations comparable to those of small individuals from the Clyde Sea Area. Comparisons between fed, unfed and plastic-fed individuals indicated a reduction in feeding rate, body mass, and metabolic rate as well as catabolism of stored lipids in plastic contaminated animals. We conclude that N. norvegicus exposed to high levels of environmental microplastic pollution may experience reduced nutrient availability. This can result in reduced population stability and may affect the viability of local fisheries.

Anticoagulant properties and cytotoxic effect against HCT116 human colon cell line of sulfated glycosaminoglycans isolated from the Norway lobster (Nephrops norvegicus) shell.

Sulfated glycosaminoglycans (SGNL) were extracted for the first time from Norway lobster (Nephrops norvegicus) shell. The monosaccharide composition analysed by GC/MS revealed the presence of galacturonic acid, glucuronic acid, N-acetylgalactosamine and N-acetylglucosamine. The analysis of SGNL with acetate cellulose electrophoresis in Zn-acetate revealed the presence of heparan sulfate (HS) and dermatan sulfate (DS). SGNL were evaluated for their anticoagulant activities using activated partial thromboplastin time (aPTT), thrombin time (TT) and prothrombine time (PT) tests. After 21h incubation, HCT116 cell proliferation was inhibited (p<0.05) between 39.7 and 54.8% at 1.5-7.5mg/mL of SGNL. SGNL don't show hemolytic activity towards bovine erythrocytes and no cytotoxicity against the normal lymphocytes. The antiproliferative efficacy of these lobster glycosaminoglycans were probably related with the higher sulfate content. SGNL demonstrated promising antiproliferative and anticoagulant potential, which may be used as a novel, effective and promising antithrombotic agent.

Bacteriostatic suppression in Norway lobster (Nephrops norvegicus) exposed to manganese or hypoxia under pressure of ocean acidification.

Future ocean acidification (OA) and warming following climate change elicit pervasive stressors to the inhabitants of the sea. Previous experimental exposure to OA for 16 weeks at pH levels predicted for 2100 has shown to result in serious immune suppression of the Norway lobster, Nephrops norvegicus. The lobsters are currently affected by stressors such as periodical hypoxia inducing high levels of bioavailable manganese (Mn). Here, we aimed to investigate possible effects of interactions between OA and these stressors on total hemocyte counts (THCs) and on recovery of inoculated bacteria in the lobsters, measured as a proxy for bacteriostatic response. The effects were judged by following numbers of culturable Vibrio parahaemolyticus in hepatopancreas, 4 and 24h post inoculation in lobsters kept in replicate tanks with six different treatments: either ambient (pCO2∼500 µatm/pH∼8.1 U) or CO2-manipulated seawater (OA; pCO2∼1550 µatm/pH∼7.6 U) for 8 weeks. During the last 2 weeks, additional stress of either hypoxia (∼23% oxygen saturation) or Mn (∼9 mg L(-1)) was added except in control treatments. Our results showed clear effect on bacteriostatic response in Norway lobsters exposed to these stressors. In lobsters kept in ambient seawater without additional stressors, the number of culturable bacteria in hepatopancreas was reduced by ∼34%. In combined treatment of ambient seawater and hypoxia, the reduction was ∼23%, while in the Mn-exposed animals, there was no reduction at all. This was also the case in all OA treatments where mean numbers of culturable V. parahaemolyticus tended to increase. In lobsters from ambient seawater with or without hypoxia, the THC was not significantly different as was also the case in OA without additional stressors. However, in OA treatments combined with either hypoxia or Mn, THC was reduced by ∼35%. While the reduction of culturable V. parahaemolyticus in lobsters was clearly affected by these stressors, we found no notable effects on growth, survival or hemolytic properties of the bacteria itself. Thus, we conclude that this predicted stress scenario is beneficial for the pathogen in its interaction with the host. As OA proceeds, it may force the health of the ecologically and economically important N. norvegicus to a tipping point if exposed to more short-term stressors such as the periodical events of hypoxia and Mn. This could impact lobster condition and biomass and may as well increase the risk for bacterial transmission to consumers.

Reproductive biology of female Norway lobster, Nephrops norvegicus (Linnaeus, 1758) Leach, in Icelandic waters during the period 1960-2010: comparative overview of distribution areas in the Northeast Atlantic and the Mediterranean.

Maturity size, reproductive cycle, sex ratio and fecundity of female Nephrops were investigated at SW, S and SE Iceland for the period 1960-2010. Time series of biological parameters and fisheries data displayed significant relationships. In addition, female biological data from 20 areas in the Atlantic and Mediterranean were compared. Fifty percentage maturity estimates had an overall range of 23.9-34.4mm CL with some anomalies in the 2000s. The reproductive cycle in Iceland has been biennial during the whole study period from mid-1960s to 2010 with minor change in phase in the 2000s. Biennial moulting retards female growth more than annual spawning, and the length of incubation and hatch time of year show significant relationships with latitude and sea temperature. Variations in sex ratio were observed and relationships found between female sex ratio and CL, CPUE and stock biomass during 1961-2010, displaying apparent fishery-induced effects on sex ratio. Potential and realized fecundity estimates in Iceland are 35-50% of those reported from more southerly waters. Biennial spawning and low fecundity limit the number of progeny in Icelandic Nephrops and necessitate lower fishing mortality. Ambient temperature in Icelandic waters has risen by 1°C since the late 1990s, generating around 30 days shorter incubation time in the 2000s, but around 3°C rise is necessary for possible annual spawning.

Comparison of the Norway lobster (Nephrops norvegicus) gut bacterial communities using 16S rDNA clone libraries and pyrosequencing.

By comparing 16S rDNA cloning and 454 pyrosequencing in the Nephrops norvegicus midgut, several common bacterial OTUs were detected. However, when only one method is to be selected, it needs to be considered whether the revealing of rare OTUs or their accurate phylogenetic relationships is mostly preferred.

A new laboratory radio frequency identification (RFID) system for behavioural tracking of marine organisms.

Radio frequency identification (RFID) devices are currently used to quantify several traits of animal behaviour with potential applications for the study of marine organisms. To date, behavioural studies with marine organisms are rare because of the technical difficulty of propagating radio waves within the saltwater medium. We present a novel RFID tracking system to study the burrowing behaviour of a valuable fishery resource, the Norway lobster (Nephrops norvegicus L.). The system consists of a network of six controllers, each handling a group of seven antennas. That network was placed below a microcosm tank that recreated important features typical of Nephrops' grounds, such as the presence of multiple burrows. The animals carried a passive transponder attached to their telson, operating at 13.56 MHz. The tracking system was implemented to concurrently report the behaviour of up to three individuals, in terms of their travelled distances in a specified unit of time and their preferential positioning within the antenna network. To do so, the controllers worked in parallel to send the antenna data to a computer via a USB connection. The tracking accuracy of the system was evaluated by concurrently recording the animals' behaviour with automated video imaging. During the two experiments, each lasting approximately one week, two different groups of three animals each showed a variable burrow occupancy and a nocturnal displacement under a standard photoperiod regime (12 h light:12 h dark), measured using the RFID method. Similar results were obtained with the video imaging. Our implemented RFID system was therefore capable of efficiently tracking the tested organisms and has a good potential for use on a wide variety of other marine organisms of commercial, aquaculture, and ecological interest.