Biochemical and histological responses in mussel (Mytilus edulis) exposed to North Sea oil and to a mixture of North Sea oil and alkylphenols

Several environmental chemicals are suspected to be responsible for adverse health effects on the reproductive system in various organisms. During this work, environmentally relevant concentrations of North Sea oil were used alone or in combination with alkylphenols and additional PAH to study the effect on vitellogenin-like protein expression and gonadal development in mussels. North Sea oil (0.5 ppm) induced the expression of phospho-proteins in both sexes indicating that some compounds are oestrogen-mimics. This induction was not seen in samples dosed with the mixture but signs of toxic effects were observed in the gonads. Indeed, numerous degenerating ovarian follicles in females and foci, similar to vertebrate melanomacrophage centres, were observed in testes.     

Molecular identification of ascaridoid nematodes from the deep-sea onion-eye grenadier (Macrourus berglax) from the East Greenland Sea

Macrourus berglax from the East Greenland Sea was studied for the presence of ascaridoid nematodes in 2001, 2002 and 2003. The fishes were collected between 278 and 413 m water depth using a benthopelagic net. Based on the amplification of the internal transcribed spacer ITS-1, 5.8S, ITS-2 and flanking sequences (=ITS+), three ascaridoid nematode species were identified. The prevalence of infestation during the 3 years ranged from 42.9% to 62.9% and 22.9% to 40.0% for the anisakids Anisakis simplex (s.s.) and Pseudoterranova decipiens (s.s.), respectively, and from 28.6% to 60.0% for the raphidascarid Hysterothylacium aduncum. A total of 18 specimens, two of each species and examination year, revealed no sibling species, suggesting a limited distribution of other ascaridoid siblings into the deep sea. The ITS-1, 5.8S and ITS-2 sequences of A. simplex (s.s.) from the East Greenland Sea did not differ from previously published sequence data (GenBank) from other regions in the Atlantic and Pacific oceans. The sequences of P. decipiens (s.s.) corresponded most closely to those of specimens from Richardson Bay, western Pacific, and differed in four positions (0.5%). They corresponded least to those of specimens from Japan (1.5%). The sequence data for H. aduncum differed in two positions in the ITS-1 (0.2%) and three positions in the ITS-2 (0.3%) from sequences from Japan. A high genetic similarity between the regions can be explained by (a) extensive final host migration in the case of A. simplex (s.s.), (b) an overlapping distribution of final host populations along the continental shelves for P. decipiens (s.s.) and (c) a low host specificity and large population size in the intermediate and final hosts for H. aduncum. The occurrence of the identified species in the macrourid fish underlines the potential of cosmopolitan ascaridoid nematodes to distribute not only horizontally but also vertically in the deep sea.     

Benzo[a]pyrene-dione-stimulated oxyradical production by microsomes of digestive gland of the common mussel, Mytilus edulis L

The potential of benzo[ a]pyrene (BaP) quinones (diones) to stimulate NAD(P)H-dependent hydroxyl radical (·OH) production (2-keto-4-methiolbutyric acid (KMBA) oxidation) by digestive gland microsomes of M. edulis was studied using iron/EDTA as a promotor of the Haber-Weiss reaction (O₂⁻ + H₂O₂ = ·OH + OH⁻ + O₂). Stimulation of basal rates of KMBA oxidation was observed for all three diones. Stimulation was similar for the 1,6- and 3,6-diones, but much less for the 6,12-dione, and greater for the NADH than the NADPH-dependent reaction, viz. % increases of 78 to 333 (NADH) compared to 0 to 78 (NADPH). Maximal KMBA oxidation was obtained at dione concentrations of 12.5 to 50 μm. Inhibition of KMBA oxidation by Superoxide dismutase and catalase indicated the involvement of respectively Superoxide anion radical (O₂⁻) and hydrogen peroxide (H₂O₂) in 1,6-dione stimulated NADPH-dependent · OH formation. The apparent K_m values of xenobiotic-stimulated KMBA oxidation were lower for BaP-diones than for the model redox cycling quinone menadione (2-methyl-1,4-naphthoquinone), viz. in μm, 0.6 to 14.6 (NADH) and 4.4 to 28.5 (NADPH) compared to 315 to 457 (NADH) and 72 to 103 (NADPH). The results are consistent with metabolism of BaP to diones and resultant enhanced generation of oxyradicals being a potential mechanism of pollutant-mediated toxicity in molluscs.     

Partial purification and properties of cytochrome P450 from digestive gland microsomes of the common mussel, Mytilus edulis L.

Cytochrome P450 from the digestive gland of M. edulis was partially purified by sodium cholate solubilization, 4–15% polyethylene glycol fractionation, and octyl-Sepharose affinity, DEAE-Sephacel ion-exchange and hydroxylapatite chromatography (yields of up to 7–10%). Three peaks were resolved by DEAE-Sephacel chromatography (termed peaks 1–3). P450 specific content was increased from 26 to 800 pmol per mg protein, and the ratio of P450 content to NADPH-cytochrome c (P450) reductase activity reduced by a factor of 250. Oxidised spectrum λmax of P450 was 410.5 ± 1.5 nm. Type II difference spectra were seen with both type II (clotrimazole, metyrapone) and type I (α-naphthoflavone, 7-ethoxycoumarin) compounds. Western blotting with polyclonal anti-P4501A from perch (Perca fluviatilis) gave a single band of approximately 54 kDa molecular weight. A reconstituted system containing peak 2 or 3, rat liver P450 reductase, lipid and NADPH metabolised benzo[a]pyrene to diones, diols, phenols and putative protein adducts. Peak 2 plus cumene hydroperoxide was indicated to produce diones and protein adducts only. Peak 2 alone was indicated to produce diones and phenols. The major free metabolites in all cases were diones (53–100%). The results indicate the existence of a P4501A-like enzyme in M. edulis, possibly with unusual properties as indicated by the difference spectra, metabolism in absence of NADPH and added P450 reductase, and predominance of diones.     

Some effects of copper on the veliger larvae of the mussel Mytilus edulis and the Scallop Pecten maximus (Mollusca, Bivalvia)

During 1983 and 1985 several batches of laboratory reared veliger larvae of Mytilus edulis and Pecten maximum here subjected to a rank of concentrations of added copper (CuCl₂) over a 15-day period. M. edulis larvae were less sensitive, measured both as mortality (15-day LC₅₀) of 400 μglitre⁻¹) and reduced growth, than P. maximus larvae (15-day LC₅₀ of 85μg litre⁻¹). Both species appeared less sensitive to Cu than other bivallve larvae previously studied. Veliger larvae of M. edulis are from 7 to 10 times more tolerant of Cu than juveniles or adults and this unexpected finding is discussed in relation to the recent literatures on Cu toxicity and accumulation in mussels.     

Biochemical and ultrastructural observations of long-term silver accumulation in the mussel, Mytilus edulis

The cellular distribution and chemical forms of Ag were determined in mussels after accumulation of the metal from sea water. The major accumulations were either in the vacuoles of connective tissue macrophages, where it was associated with S, or in deposits in the fibrillar layer of the basement membranes of the digestive diverticulum and kidney epithelia, where it was bound to the sulphydryl and sulphate groups of glycoproteins and proteoglycans. In acutely exposed animals, about 10 % of the accumulated Ag was found in a (Ag, Cu)-binding protein with characteristics indicating that it may be a metallothionein. There was a concomitant increase in the body Cu levels after Ag exposure and 40 % of this was bound to this low molecular weight metal-binding protein.     

Recruitment variability in dab (Limanda limanda) in the southeastern North Sea

Data from three annual surveys, covering inshore and offshore waters of the southeastern North Sea, were analysed to study recruitment variability in dab (Limanda limanda) over the period 1978–1997. Geometric mean abundance of 0- to 5-group dab was estimated using general linear models. Juvenile dab (0- and 1-group) were found over the entire area, from inside the estuaries to 50 m depth offshore. Environmental conditions (water temperature, wind stress, turbidity) affected the catch rates. The potential errors in the estimates of year-class strength, caused by differences in catchability, are discussed. The inter-annual pattern of year-class strength appeared to be established between ages 1 and 2, suggesting that factors determining recruitment are not restricted to the pelagic early life phase only, but also operate during the demersal juvenile phase. Recruitment variability at age 2 was in the order of 50–60% and appears to be equal to, or lower than, recruitment variability in plaice and sole. These results contradict expectations based on the concentration hypothesis, which states that the degree of variation in recruitment is inversely related to the degree of concentration during early life phases.     

Ab initio investigations of the charge transport properties of endohedral M@C20 (M= Na and K) metallofullerenes

Using density functional theory and quantum transport calculations based on nonequilibum Green's function formalism, we investigate the charge transport properties of endohedral M@C₂₀ (M= Na and K) metallofullerenes. Our results show that the conductance of C₂₀ fullerene can be obviously improved by insertion of alkali atom at its centre. Both linear and nonlinear sections are found on the I-V curves of the Au-M@C₂₀-Au two-probe systems. The novel negative differential resistance behaviour is also observed in Na@C₂₀ molecule but not in K@C₂₀.     

Bioaccumulation of heavy metals by bivalves from Limski Kanal (North Adriatic Sea). II. Copper distribution between oysters, Ostrea edulis, and ambient water

Accumulation of copper was studied between June 1979 and June 1980 in the oyster Ostrea edulis Linnaeus, cultivated in the Limski Kanal, on the Istria Peninsula (Yugoslavia).The copper distribution in different organs of oysters of the same age was compared with concentrations of copper existing in different physicochemical forms in ambient sea water, i.e., dissolved Cu in labile complexes determined at the natural pH of 8; ‘total’ dissolved Cu determined after acidification to pH 2; and Cu bound to suspended particles with a diameter above 0.45 μm. Copper concentrations in the water were determined by differential pulse anodic stripping voltammetry (DPASV) and in analyses of biotic material, graphite furnace atomic absorption spectrometry (GFAAS) was applied.Significant correlations have been found between copper complexed with dissolved organic ligands and its concentration in the soft part of the oyster. Copper contents increase gradually with the weight of the organs until enhanced gametogenetic activity of the oysters occurs. Thereafter the correlation factor between copper contents and the size of body parts varied significantly with the season.     

三种典型POPs对紫贻贝不同组织DNA损伤的比较研究

采用彗星电泳技术,比较研究了Aroclor 1254、BaP、DDT三种典型POPs对紫贻贝(Mytilus edulis)鳃、性腺、消化腺细胞的DNA损伤效应的差异,以期为进一步评价这三种典型POPs遗传毒理机制及其预警监测提供科学依据.结果表明,Tail DNA%是评价DNA损伤的理想指标,另外,在急性毒性试验中,...     

Effects of long-term exposure to silver or copper on growth, bioaccumulation and histopathology in the blue mussel Mytilus edulis

The purpose of this study was to determine the long-term accumulation of either silver or copper from low concentrations in seawater by blue mussels, Mytilus edulis. Mussels raised from eggs in the laboratory to the age of 2·5 months (approximately 4·5 mm in length) were continuously exposed to 0, 1, 5 and 10μg/liter of either silver (nitrate) or copper (chloride) and sampled at 12, 18 and 21 months for growth studies, measurements of metal accumulation and histopathological examination.Whole-body soft tissues were analyzed for the presence of both silver and copper, as background levels of copper in the incoming seawater averaged 2–4 μg/liter. Mussels exposed to silver had accumulated significant amounts of silver only at the highest test concentration (10 μg/liter Ag) after 12 months, but at 18 and 21 months significant levels were accumulated at all three test concentrations. Mussels exposed to copper accumulated significant amounts of copper at 5 and 10 μg/liter Cu after all three sampling periods, but not at 1μg/liter. Silver-exposed animals also accumulated significantly greater amounts of copper than control animals.In a comparative study, field-collected juvenile mussels (approximately 16·1 mm in shell length) and adult mussels (approximately 53·4 mm in shell length) were exposed for 12 months to 0, 5, 25 and 50 μg/liter silver only and subsequently sampled for metal-accumulation analyses and growth measurements. Juvenile mussels accumulated significant amounts of silver at all test concentrations, with the exception of mussels exposed to 5 μg/liter Ag for 6 months. Copper accumulation in the silver-exposed juveniles was significant only at 50 μg/liter Ag after 6 months, but at all test concentrations after 12 months. Adult mussels exposed to silver accumulated significant levels of both silver and copper, but at somewhat lower levels than juveniles.In the growth study, silver had no effect on laboratory reared mussels at the highest concentration of 10 μg/liter tested, whereas copper at 10 μg/liter did appear to affect growth as early as 4 months after the start of experimental exposure. Field-collected juvenile mussels did show inhibition in growth after 6 months' exposure to 25 and 50 μg/liter Ag, with some growth occurring after 12 months. Adults also showed inhibition in growth after 6 months but not at 12 months.Histopathological examination of mussels exposed to either 5 or 10 μg/liter of copper for 18 months showed changes in the digestive diverticula, gastrointestinal tract, reproductive tract and muscle tissues. These changes were more noticeable in mussels exposed to 5 μg/liter Cu than in those exposed to 10 μg/liter. Mussels exposed to silver for 21 months showed yellowish to black particulate deposition in the basement membrane and connective tissue of the various organs and tissues. Silver deposition increased with increasing test concentration.     

A new meiofauna polychaeta Pholoe (Polychaeta, Sigalionidae)from the Huanghai Sea (Yellow Sea)

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The impact of coastal defence structures (tetrapods) on decapod crustaceans in the southern North Sea

Although the use of coastal defence structures is expected to increase, little is known about the ecological impact of such structures on the natural environment. In particular, the temporal and spatial patterns of communities in association with artificial substrate are still poorly understood. This study examined possible effects of experimental tetrapod fields on the decapod crustacean community in a subtidal hard-bottom area in the southern North Sea. We performed in situ studies in the fields and along transects oriented away from the tetrapod fields. Species composition and abundances were assessed before and after the introduction of the artificial material. The study revealed a significant decrease of smaller, less vagile species (Pisidia longicornis, Pilumnus hirtellus, Galathea squamifera) over the entire study area in the years following the tetrapod introduction. For 2 species, Hyas araneus and Homarus gammarus, the tetrapods appeared to be highly attractive as habitat and shelter because their abundance increased over time. No distinct spatial or temporal effects were observed for mobile predatory crabs, such as Cancer pagurus and Liocarcinus spp. The results of the study demonstrate that possible effects of artificial structures on macro-invertebrates in temperate hard-bottom areas are highly species-specific and depend on the size, lifestyle and ecological requirements of the species. This work highlights the importance of long-term studies. Our findings clearly indicate that more time is needed to distinguish between natural and anthropogenic influences on species distributions.     

Relationships between daily growth of different groups of swordtip squid (Uroteuthis edulis) and environmental variables in the East China Sea

Swordtip squid (Uroteuthis edulis) is one of the important economical fishing target species in the East China Sea. Uroteuthis edulis is characterized by rapid growth, extensive migration, and long spawning period and sensitive to surrounding environment. In order to assess its stock status, it is necessary to explore its spawning season, growth patterns of different populations and their relationship with the environment in advance. In this paper, based on the samples of U. edulis collected in the East China Sea from September 2017 to March 2018, we explored the relationships between daily growth of statolith microstructure and environmental variables by gradient forest method and generalized additive model. The spawning season of U. edulis was found to be nearly one year, and two dominant season groups were found: the spring group with the peak period of April and the summer group with the peak period of August. Water temperature in the depth of 25 m (Temp_25), sea surface temperature (SST) and zonal velocity were the key environmental variables for the daily growth of April-spawning group. The most suitable environmental conditions for the growth of April-spawning group were water temperature (24?27°C) and velocity (0.1?0.3 m/s). SST, Temp_25 and mixed layer depth were the key environment variables for the daily growth of August-spawning group. The most suitable environmental conditions for the growth of August-spawning group were water temperature (21?28°C) and water depth (0?50 m). Key environmental variables of different groups suggested that early growth was significantly affected by seasonal changes of water temperature, current velocity and prey abundance. This study explored the relationships between early growth and environmental variables and provided the scientific guidance for the management and conservation of U. edulis.     

Development of habitat models for Nephtys species (Polychaeta: Nephtyidae) in the German Bight (North Sea)

The aim of the study was the development of habitat models for Nephtys species (Polychaeta: Nephtyidae). The investigation area was the German Bight, the southeastern part of the North Sea. Models were developed based on field data collected between 2000 and 2006. In addition, data on environmental variables were retrieved from long-term monitoring data sets and from the sediment map by Figge [Figge, K., 1981. Nordsee. Sedimentverteilung in der Deutschen Bucht. Map No. 2900. Publisher: Deutsches Hydrographisches Institut, Hamburg]. The statistical modelling technique used was multivariate adaptive regression splines (MARS). Models were fitted individually for each species. Evaluation of predictive discrimination and predictive accuracy of the developed models was by calculation of the area under the receiver operating curve (AUC) or sensitivity and specificity, respectively. Habitat models with best predictive fit were selected for the presentation of habitat suitability maps.Six Nepthys species were found: Nephtys assimilis, N. caeca, N. cirrosa, N. hombergii, N. incisa and N. longosetosa. N. hombergii was most common whereas N. incisa and N. longosetosa were rare. Habitat preferences varied considerably among the species. For all investigated Nephtys species except N. longosetosa a habitat model could be developed based on four predictor variables. The habitat models with best predictive fit were those for N. cirrosa and N. hombergii. The N. caeca habitat model was of limited predictive accuracy and only accept predictive discrimination. The number of predictors as well as the relative importance of the respective predictors in the model varied among the different species. Direct comparison of most suitable habitats for the different species based on modelling revealed that in the mostly sandy regions parallel to the German coast in water depths up to 20 m an overlap between N. caeca, N. hombergii and N. cirrosa exists. In the deeper central German Bight with mostly fine sands with increased mud contents N. hombergii, N. assimilis and, at least partially and rare in numbers, N. incisa co-occur. It can be concluded that important sediment characteristics like grain size median and mud content as well as water depth and mean salinity are useful parameters to describe the habitat requirements of most Nepthys species in the German Bight. However, additional variables need to be incorporated into such analyses.     

Effects of linear alkylbenzene sulphonate (LAS) and cadmium in the digestive gland of mussel, Mytilus sp.

A laboratory study was carried out exposing mussels (Mytilus sp.) to linear alkylbenzene sulphonate (LAS) (6 mg litre⁻¹), Cd (0.05 mg litre⁻¹) and LAS plus Cd at the same concentrations. The aim was to assess the use of several histopathological and biochemical indices as potential biomarkers of the impact of these xenobiotics in the digestive gland of molluscs. Treated mussels actively accumulated Cd in the digestive gland compared with controls (p 0.01), the highest levels occurring after 30 days of exposure in the group treated with Cd plus LAS. Among several histological alterations screened in digestive gland tissues the thickness of digestive tubules in Cd treated animals decreased more markedly (p 0.01) than in LAS exposed mussels. As for biochemical parameters, the investigated antioxidant enzyme activities, superoxide dismutase (SOD), catalase, DT-diaphorase and glutathione peroxidase (GPX) did not show any significant induction due to these xenobiotics. However, a slight decrease of the antioxidant defences of the animals was detected after 30 days of exposure to contaminants.