Crustacean Nephrops Norvegicus Research Articles

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 pilot study for an innovative approach highlighting Actin and COI mRNAs as potential biomarkers of quality of the edible crustacean Nephrops norvegicus (Linnaeus, 1758)

Widely appreciated in gastronomy, many harvested and farmed Crustacean species are a great economic resource in the fishery business, throughout the world. Factors like pollutants and/or improper management in on-board/on-shore practices may adversely affect their edible quality and shelf-life. Recent studies from applied scientific research have been supporting the fishing industry and trading companies, in order to improve post-harvest practices and offer a safe and competitive product of high quality. Here, we developed an innovative molecular approach to get basic information on crustacean quality by using the European lobster Nephrops norvegicus (Linnaeus, 1758). For the first time, we investigated muscle-exudates of thawed lobsters for the presence of nucleic acids and analysed mRNA targets selected at NCBI to test their potential use as biomarkers of quality in frozen-thawed seafood. By RT-PCRs and sequencing we validated small amplicons of Actin, Cytochrome oxidase subunit 1 (COI), Calpain M, Prophenoloxidase (ppo) and Crustin-like antimicrobial peptide. In comparative One Step RT-PCR and/or One Step RT-qPCR assays, we analysed their amplification profiles at 1, 4 and 7 days after thawing (a.t.), in lobsters maintained to checked temperatures from capture. Based on amplification efficiency (AE), inversely correlated to the time after thawing and loss of RNA targets integrity, we identified new highly sensitive, species specific early indicators of quality. Our results highlight the advantage of using amplicons from exudates in different food science applications including authenticity/traceability, and provide new insights for the development of scientific tools applicable to crustacean quality monitoring on industrial-scale.

Evidence for the decline and possible extinction of a marine parasite species caused by intensive fishing

Abstract The first good evidence of the role of intensive commercial fishing in the decline and possible extinction of a marine parasite is presented. The larva of Stichocotyle nephropis Cunningham, 1887 (Aspidogastrea: Digenea) was described in 1887 from the outside of the intestine of the decapod crustacean Nephrops norvegicus (L., 1758) in the North Sea. Similar larvae were found outside the intestine of the American lobster Homarus americanus (Milne-Edwards, 1837) caught off New England in 1895. The adult form was first found in 1898 in the bile ducts of the thornback ray Raja clavata (L., 1758) from the North Sea, then in the barndoor skate Dipturus laevis (Mitchill, 1818) caught off Nantucket. There have been few records of its occurrence since then, with the last records from the northeast and northwest Atlantic published in 1972 and 1986 respectively, despite the recent attempts reported herein to find larvae and adult parasites. Both the elasmobranch final hosts are listed as “near threatened” or “ endangered” and the latest evidence on their population sizes suggests that they may have been fished to below the thresholds required for successful transmission of the parasite.

Managing Marine Plastic Pollution: Policy Initiatives to Address Wayward Waste

A few times a year, volunteers fan out along the causeway that links the South Carolina mainland with the seashore community of Folly Beach to clean up plastic bottles, straws, bags, and other debris from along the road and the salt marsh. Some of this debris has come from cities miles away. On windy days, litter is often blown off city streets into waterways. During rainstorms, debris floats into drains that empty into rivers. Other trash probably came from places closer to home. “I see bags and other plastic flying off the beds of pickup trucks going down the causeway,” says Marty Morganello, who organizes the cleanups for the Charleston-area chapter of the nonprofit Surfrider Foundation. “I see them coming out the open windows of cars and out the backs of garbage trucks and even recycling trucks. This material is lightweight, and if you don’t secure it, it will fly away.” By one estimate, the volume of plastic debris going into the world’s oceans could more than double by 2025, assuming current trends in coastal development and plastics use. Some countries have begun identifying ways to improve management of plastic ... Beach cleanups yield enormous amounts of trash, with plastic items a major constituent.1 Although the human health impacts of this marine plastic pollution remain poorly characterized, it is widely seen as an emerging problem that deserves much more research attention.2 Likewise, there is a growing urgency among industry, government, nongovernmental organizations, and environmental groups to develop tools and policies to track, capture, and recycle plastic waste before it reaches the ocean.

Behaviour influences larval dispersal in shelf sea gyres: Nephrops norvegicus in the Irish Sea

The western Irish Sea seasonal gyre is widely believed to play an important role in the local retention of resident larvae. This mechanism could be particularly crucial for the larvae of the heavily fished crustacean Nephrops norvegicus (L.), as their sediment requirements highly restrict where they are able to settle. As recent research suggests that the gyre may be becoming less retentive due to changes in atmospheric forcing, it is now crucial to understand how the gyre influences dispersal. This investigation addresses the hypothesis that shelf sea gyres reinforce larval retention using a biophysical model with vertical migration, habitat selection and temperature-dependent pelagic larval duration (PLD) configured to match the behaviour of N. norvegicus larvae. The results of this study suggest that the gyre does increase the likelihood that passive larvae remain within the western Irish Sea, on the condition that the larvae remain fixed at the depth of peak gyral flow. Retention rates are significantly lower when vertical migration is introduced, and there is no evidence that the gyre promotes larval retention amongst either vertically migrating larvae, or larvae that require muddy sediments for successful settlement. By contrast, vertical migration is shown to be favourable for retention in the eastern Irish Sea. PLD varies by a factor of two according to release date and location. The simulations suggest that whilst some highly limited and almost entirely unidirectional larval exchange may occur, the distinct sites largely rely upon local recruitment.

Simulated climate change causes immune suppression and protein damage in the crustacean Nephrops norvegicus

Rising atmospheric carbon dioxide concentration is causing global warming, which affects oceans by elevating water temperature and reducing pH. Crustaceans have been considered tolerant to ocean acidification because of their retained capacity to calcify during subnormal pH. However, we report here that significant immune suppression of the Norway lobster, Nephrops norvegicus, occurs after a 4-month exposure to ocean acidification (OA) at a level predicted for the year 2100 (hypercapnic seawater with a pH lowered by 0.4 units). Experiments carried out at different temperatures (5, 10, 12, 14, 16, and 18°C) demonstrated that the temperature within this range alone did not affect lobster immune responses. In the OA-treatment, hemocyte numbers were reduced by almost 50% and the phagocytic capacity of the remaining hemocytes was inhibited by 60%. The reduction in hemocyte numbers was not due to increased apoptosis in hematopoetic tissue. Cellular responses to stress were investigated through evaluating advanced glycation end products (AGE) and lipid oxidation in lobster hepatopancreata, and OA-treatment was shown to significantly increase AGEs', indicating stress-induced protein alterations. Furthermore, the extracellular pH of lobster hemolymph was reduced by approximately 0.2 units in the OA-treatment group, indicating either limited pH compensation or buffering capacity. The negative effects of OA-treatment on the nephropidae immune response and tissue homeostasis were more pronounced at higher temperatures (12–18°C versus 5°C), which may potentially affect disease severity and spread. Our results signify that ocean acidification may have adverse effects on the physiology of lobsters, which previously had been overlooked in studies of basic parameters such as lobster growth or calcification.

Effects of manganese and hypoxia on coelomocyte renewal in the echinoderm, Asterias rubens (L.)

Manganese (Mn) is a naturally abundant metal and particularly so in soft-bottom oceanic sediments where it generally occurs bound in a four-valent colloidal state as MnO 2. When hypoxic conditions occur in bottom waters, the metal reduces to the bioavailable ion Mn 2+ and can reach concentrations known to have immunotoxic effects in the crustacean Nephrops norvegicus, reducing numbers of circulating haemocytes as a consequence. However, we have previously shown that Mn seems to have a contrasting effect on the echinoderm Asterias rubens in which it triggers the proliferation of haematopoietic cells and increases coelomocyte numbers. Since elevated Mn levels mostly co-occur with hypoxia in nature, here we investigated whether hypoxia has a negative effect on haematopoiesis. Proliferation and differentiation of coelomocytes and cells in the coelomic epithelium of A. rubens were compared after 3 days of exposure to realistic levels of Mn, hypoxia or a combination of these two parameters. We can confirm that Mn elevated numbers of coelomocytes and increased proliferation of epithelial cells, but hypoxia did not affect these levels. However, hypoxia did affect differentiation of these cells as judged by investigating the expression of a Runt domain transcription factor, which was also cloned and sequenced. Through comparative quantification using a real time PCR technique, we found that exposure to hypoxia had a clearly stimulating effect on mRNA expression of Runt gene in both coelomocytes and epithelial cells. These results indicate that during hypoxic conditions the composition of coelomocyte sub-populations changed.

The influence of light availability and predatory behavior of the decapod crustacean Nephrops norvegicus on the activity rhythms of continental margin prey decapods

AbstractThe day–night cycle is one of the strongest geophysical cycles modulating species’ behavioral rhythms. However, in deep‐water continental margins, where light intensity decreases over depth, interspecific competition may alter behavioral responses to day–night cycles. The burrowing decapod crustacean Nephrops norvegicus is a large‐size predator in benthic communities, exerting despotic territorial behavior. In this study, we analysed how the effect of light intensity cycles on decapod behavioral rhythms is reduced as one moves from shelves to slopes. In the Western Mediterranean, the predatory behavior and interspecific competition for substrate use of Nephrops increases moving from the shelf (100–110 m) to the slope (400–430 m). Vector fitting and generalized additive models were used to assess the effect of light intensity and behavioral rhythms of N. norvegicus on the temporal variation of prey decapods co‐occurring in trawl tow catches carried out on the shelf and the slope during October 1999 and June 2000. The combination of diel variations in light intensity and N. norvegicus abundance influences the activity rhythms of prey decapods in a depth‐ and seasonal‐dependent manner. Light modulation is stronger on the shelf and weaker on the slope, where Nephrops population size is greater. Although present regression analysis does not necessarily imply a direct cause–effect relationship between rhythms of predators and prey, we suggest that Nephrops alters the temporal patterning in the behavior of its prey on the slope, where light intensity is reduced. This alteration is stronger in endobenthic species than in benthopelagic species; the former rely on bottom substrate for the expression of behavioral rhythms, experiencing stronger interspecific competitions with Nephrops at time of activity.

A comparative study on the influence of manganese on the bactericidal response of marine invertebrates

Manganese, Mn, is a naturally abundant metal in marine sediments. During hypoxic conditions the metal converts into a bioavailable state and can reach levels that have been shown immunotoxic to the crustacean Nephrops norvegicus. For this species it has previously been shown that exposure to 15 mg L −1 of Mn decreased the number of circulating haemocytes while it for the echinoderm Asterias rubens increased the number of coelomocytes. Here, we compared if five days of exposure to the same concentration of Mn affects the bactericidal capacity of these two species and the mollusc Mytilus edulis when inoculated with the bacterium Vibrio parahaemolyticus. Viable counts of the bacteria were investigated at a time-course post-injection in the blood and the digestive glands of Mn-exposed and un-exposed (controls) animals. Accumulation of Mn was also analyzed in these tissues. When exposed to Mn the haemocyte numbers were significantly reduced in M. edulis and it was shown that the bactericidal capacity was impaired in the mussels as well as in N. norvegicus. This was most obvious in the digestive glands. These two species also showed the highest accumulation of the metal. In A. rubens the bactericidal capacity was not affected and the metal concentration was similar to the exposure concentration. After a recovery period of three days the concentration of Mn was significantly reduced in all three species. However, in M. edulis and N. norvegicus it was still double that of A. rubens which could explain the remaining bactericidal suppression observed in N. norvegicus. This study pointed out that exposure to such Mn-levels that are realistic to find in nature could have effects on the whole organism level, in terms of susceptibility to infections. The effect seemed associated to the accumulated concentration of Mn which differed on species level.

Day–night and depth differences in haemolymph melatonin of the Norway lobster, Nephrops norvegicus (L.)

Few studies have been conducted to quantify and understand the role of melatonin in invertebrates, and particularly in crustaceans and in deep-sea animals. In this study, we examined day–night differences in haemolymph melatonin of the burrowing decapod crustacean Nephrops norvegicus (L.) during exposure to cycles of monochromatic blue light (480 nm) and darkness cycles of 10 and 0.1 lx. These differential intensity conditions simulate illumination at the depth of the shelf (80–100 m) and of the slope (300–400 m), where these lobster populations are chiefly found in the Western Mediterranean Sea. Our objectives were: (a) to verify the presence of melatonin in the haemolymph of this species using liquid chromatography/tandem mass spectrometry (LC–MS/MS) and fluorescence HPLC (HPLC); and (b) to study the relationship between diel variations in melatonin concentration and locomotor rhythms, in order to examine whether the former influences behaviour. Melatonin was identified in LC–MS/MS by Q1 and Q3 mass peaks at an elution time of 3.7 min, and it was also detected by HPLC. Melatonin concentration was found to be two orders of magnitude higher at 10 lx (4.8±5.3 ng ml −1) than at 0.1 lx (0.06±0.03 ng ml −1). Also, the increase at daytime in 10 lx was absent in 0.1 lx. When the locomotor rhythm of animals exposed to both photoperiod regimes was compared, the diel periodicity was found to be preserved, but the timing of activity shifted from night to day. Extrapolating these data to the field, we interpret our results to mean that locomotor activity preserves its diel character, but not its phase and amplitude, in a bathymetric range where haemolymph melatonin reduces its concentration and rhythmic fluctuation.

The accumulation and retention of 95mTc by the Norway lobster, Nephrops norvegicus L.

Laboratory experiments were carried out to study bioaccumulation and determine a concentration factor (CF) for technetium ( 95mTc) in the homarid crustacean Nephrops norvegicus L. The steady state CF for accumulation from seawater was estimated to be about 2000 and the biological half-time was about 50 days. The highest tissue Tc concentrations were found in the green gland and the digestive gland. Depuration following accumulation from water was slow with a half-time of about 165 days. Tc accumulation from labelled food followed a biphasic model with one compartment containing about 94 percent of the ingested activity and with a half-time of about 1 day and the second compartment containing about 6 percent of the ingested activity with a half-time of about 56 days. Most retained activity was found in the digestive gland.

Functional properties of a hybrid vacuolar H(+)-ATPase in Saccharomyces cells expressing the Nephrops 16-kDa proteolipid.

The hydrophobic 16-kDa polypeptide which forms gap-junction-like structures in the crustacean Nephrops norvegicus is a member of a highly conserved family of proteolipids involved in a variety of membrane transport functions in eukaryotic cells. This family also includes the product of the Saccharomyces cerevisiae VMA3 gene which encodes an integral membrane component of the vacuolar membrane H(+)-ATPase. The cDNA for the Nephrops proteolipid complements a mutation in the yeast VMA3 gene, resulting in assembly of a hybrid H(+)-ATPase comprising yeast catalytic subunits and Nephrops integral membrane components. The hybrid vacuolar ATPase was capable of ATP hydrolysis which was coupled to proton translocation and showed inhibitor binding and enzymological properties similar to those of wild-type V-ATPases (Km for ATP, 0.4 mM), suggesting that both yeast and crustacean proteolipids share conserved structure at regions of protein interaction. To facilitate isolation of the Nephrops proteolipid by affinity chromatography on a Ni(2+)-binding support, six C-terminal histidine residues were added to the proteolipid. This modification did not prohibit assembly into the hybrid H(+)-ATPase, although the resultant enzyme did have a markedly elevated Km (1.8 mM). The membrane-bound Vo sector of the ATPase was isolated by the affinity-chromatography procedure and reconstituted into synthetic vesicles. This complex was found to be impermeable to small cations in the absence of catalytic ATPase subunits either in situ in the vacuolar membrane or in the reconstituted system. The functional significance of this impermeability and the structure/function relationships between proteolipids from different sources are discussed.

Structure and biological activity of crustacean gastrointestinal peptides identified with antibodies to gastrin/cholecystokinin

Four gastrin/cholecystokinin-like peptides (G/CCK) which cross-react with a specific C-terminal gastrin/CCK antiserum have been isolated from the stomach of the marine crustacean Nephrops norvegicus. The molecular weight of the four peptides was estimated between 1000 and 2000 Da by molecular sieving. By radioimmunoassay, the cross-reactivity of these peptides with human gastrin 17-I was found to be around 0.03%. Pure peptidic fractions were recovered after four successive steps of HPLC. Amino-acid analysis suggested a similarity between the four peptides identified which may belong to a new family. A limited homology between the C-terminus of one Nephrops peptide and vertebrate G/CCK was found after sequenceing. Two of the peptides exhibited secretagogue effects on crustacean isolated midgut glands. The Nephrops peptides, although structurally distinct from the vertebrate G/CCKs, appear to serve similar biological functions in crustaceans.

Calcitonin gene-related peptide immunoreactivity in a crustacean Nephrops norvegicus and correlation with calcitonin

Immunoreactive calcitonin-gene-related peptide (ir-CGRP) was detected in the crustacean Nephrops norvegicus. High levels of ir-CGRP were present in the foregut and hepatopancreas (3 ± 0.7 and 4.6 ± 1.0 μg eq per 100 mg of fresh organ, respectively). Molecular sieving of acidic extracts of anterior gut of Nephrops norvegicus showed a high molecular weight immunoreactive peptide in the range 15,000 to 25,000 Da. Immunoreactivity related to salmon calcitonin was present in the high molecular weight fraction.

The gap junction-like form of a vacuolar proton channel component appears not to be an artifact of isolation: An immunocytochemical localization study

Gap junctional structures containing a 16-kDa intrinsic membrane protein have been isolated from the hepatopancreas of the crustacean Nephrops norvegicus. These structures are double membranes 14–15 nm thick and composed of hexagonal arrays of particles which have a central pore that is penetrated by a cationic negative stain. Membrane preparations have also been isolated from the hepatopancreas and these contain similar gap junctional regions of uniform width. Affinity purified antibodies to the 16-kDa protein bind principally to these gap junctional regions. Antiserum raised against the isolated gap junctional structures binds strongly to the lateral surfaces of the columnar epithelial cells and in particular to gap junction-like regions.

Detection of unconjugated and conjugated steroids in the ovary, eggs, and haemolymph of the decapod crustacean Nephrops norvegicus.

Gas chromatography/mass spectrometry with selected ion monitoring has been employed to examine extracts from the ovary, eggs, and haemolymph of the marine prawn, Nephrops norvegicus, to demonstrate the presence of steroids. Both free and conjugated steroids were isolated by solvent partitioning and chromatography (lipophilic Sephadex, reversed-phase Sep Pak, and normal phase medium-pressure liquid chromatography) and steroidal conjugates were cleaved enzymatically. Steroids were determined as their methyloxime derivatives, trimethylsilyl (TMS) ethers or methyloxime-TMS ethers. All assignments were based on the detection of characteristic ions and cochromatography with the authentic steroid derivatives. 5 alpha-Dihydrotestosterone, testosterone, pregnenolone, and 20 alpha-hydroxypregn-4-en-3-one were detected in unconjugated form in the ovary. The eggs and haemolymph were found to contain unconjugated 17 beta-estradiol. Conjugated 5 alpha-dihydrotestosterone was detected in both the ovary and haemolymph, but no conjugated steroids were found in the eggs.