Integrated assessment of biological responses to pollution in wild mussels (Mytilus edulis) from subarctic and arctic areas in the Norwegian sea.

North Atlantic and Arctic Oceans contain large amount of undiscovered oil and gas reserves. Therefore threat of oil spills and its hazardous ecological consequences are of great importance to the marine environment. Although mussels (Mytilus sp.) respond clearly to contaminants, biomarkers have shown variability linked to biological and environmental changes. In order to help avoiding misinterpretation of biological responses the aim of this study was to reveal the effect of natural variability in the responsiveness to pollution of a battery of cell and tissue-level biomarkers in mussels. Mussels were collected in relatively non-impacted and potentially impacted sites at ports and the vicinity of a waste water treatment plant in Trondheim and Tromsø in autumn of 2016. Although the battery of biomarkers used herein proved to be useful to discriminate impacted and non-impacted mussel populations, some confounding factors altering the biological responses were identified. Geographical/latitudinal factors seemed to be critical regarding the reproductive cycle, reserve material storage and the prevalence of parasites such as Gymnophallus cf. Bursicola trematodes. Mussels from the reference site in Tromsø displayed general stress responses at different levels, which could be influenced by the pathogenic effect of the Gymnophallus cf. Bursicola trematode and by a more advanced gametogenic developmental stage compared to the mussels from Trondheim, which could lead to misinterpretation of the reasons behind the measured stress levels in those mussels. Despite these confounding effects, the use of integrative tools such as IBR index helped to discriminate mussel populations from chemically impacted and non-impacted sites. Overall, this work serves as an anchor point both as a reference of the baseline level values of the analyzed endpoints in the studied geographical area and time of the year, and as an indication of the potential extent of the environmental confounding factors in monitoring programs causing stress on the analyzed mussel populations.

A simulated toxic assessment of cesium on the blue mussel Mytilus edulis provides evidence for the potential impacts of nuclear wastewater discharge on marine ecosystems.

The toxic effects of cesium (Cs) on the blue mussel Mytilus edulis were experimentally investigated to assess the potential environmental consequences of the discharge of nuclear wastewater containing radionuclides. A simulated experimental system of stable cesium (133Cs) was set up to mimic the impacts of radiocesium, and its heavy metal property was emphasized. The mussels were exposed to a concentration gradient of 133Cs for 21 days, followed by another 21-day elimination period. 133Cs exposure resulted in effective bioaccumulation with distinct features of concentration dependence and tissue specificity, and hemolymph, gills and digestive glands were recognized as the most target tissues for accumulation. Although the elimination period was helpful in reducing the accumulated 133Cs, the remaining concentrations of tissues were still significant. 133Cs exposure presented little effect on growth status at the individual level but had distinct interference on feeding and metabolism indicated by the oxygen consumption rate, ammonia-N excretion rate and O:N ratio, simultaneously with the impairment of digestive glands. Regarding hemocytes in the hemolymph, the cell mortality increment, micronucleus promotion, lysosomal membrane stability disruption and phagocytic ability inhibition suggested that the immune function was injured. The cooccurrence of reactive oxygen species overproduction had a close relationship with the observed damages and was thought to be the possible explanation for the immune toxicity. The assay based integrated biomarker response (IBR) presented a good linear relation with the exposure concentrations, suggesting that it was a promising method for assessing the risk of 133Cs. The results indicated that 133Cs exposure damaged M. edulis at the tissue and cell before at the macroscopic individual, evidencing the potentially detrimental impacts of nuclear wastewater discharge on marine ecosystems.

Integrated biomarker assessment of the effects of tailing discharges from an iron ore mine using blue mussels (Mytilus spp.).

The blue mussel (Mytilus spp.) has been used to assess the potential biological effects of the discharge effluent from the Sydvaranger mine, which releases its tailings into Bøk fjord at Kirkenes in the north of Norway. Metal bioaccumulation and a suite of biomarkers were measured in mussels positioned for 6 weeks at varying distances from the discharge outlet. The biomarkers used included: stress on stress (SS); condition index (CI); cellular energy allocation (CEA); micronuclei formation (MN); lysosomal membrane stability (LMS), basophilic cell volume (VvBAS); and neutral lipid (NL) accumulation. The individual biomarkers were integrated using the integrated biological response (IBR/n) index. The accumulation of Fe was significantly higher in mussels located closer to the discharge outlet, indicating that these mussels had been exposed to the suspended mine effluent. The IBR/n results were in good agreement with the location of the mussels in relation to the distance from the discharge outlet and expected exposure to the mine effluent. Several biomarkers showed responses resulting in higher IBR/n values of analysed mussels within a 3 km distance from the tailing discharge.

Mussels as a tool for mitigation of nutrients in the marine environment.

Long-line mussel farming has been proposed as a mitigation tool for removal of excess nutrients in eutrophic coastal waters. A full-scale mussel farm optimized for cost efficient nutrient removal was established in the eutrophic Skive Fjord, Denmark where biological and economic parameters related to nutrient removal was monitored throughout a full production cycle (1 yr). The results showed that it was possible to obtain a high area specific biomass of 60 t WW ha(-1) eqvivalent to a nitrogen and phosphorus removal of 0.6-0.9 and 0.03-0.04 t ha(-1)yr, respectively. The analysis of the costs related to establishment, maintenance and harvest revealed that mussel production optimized for mitigation can be carried out at a lower cost compared to mussel production for (human) consumption. The costs for nutrient removal was 14.8 € kg(-1)N making mitigation mussel production a cost-efficient measure compared to the most expensive land-based measures.

Assessment of biomarkers in Mytilus edulis to determine good environmental status for implementation of MSFD in Ireland.

Candidate OSPAR/ICES recommended biomarkers at the level of the individual in Mytilus edulis for determination of good environmental status for MSFD were evaluated against contaminant levels at sites around Ireland. The sites chosen ranged from moderate to low pollution levels, but the actual ranking of the sites varied according to the contaminant levels present. At the most contaminated site, Cork, 4 out of 16 contaminants exceeded the EAC, while at Shannon, no EACs were exceeded. The SOS assay suggested that Cork was the healthiest site with a LT50 of 17.6 days, while SOS for Shannon was 15.6 days. Likewise, condition factors varied among sites and did not always correspond to contaminant-based status. There may be uncertainty in assigning status around the not good:good boundary. This raises potential difficulties not only in the biomarker/contaminant load relationship but also in the reliability of the biomarkers themselves and hence barriers meeting compliance levels.

Assessment of sacrificial anode impact by aluminum accumulation in mussel Mytilus edulis: a large-scale laboratory test.

Since the early 1960s, the application of aluminum alloy sacrificial anodes to mitigate marine corrosion has been well known. The aim of this work was to study aluminum bioconcentration in Mytilus edulis by an in vitro test performed in two tanks: the first containing non-contaminated water (NCW) and the second containing aluminum-contaminated water (CW) (530 µg L(-1)) released by sacrificial anode. The mussels were collected and examined over a period of 8 weeks. A comparison between the aluminum concentrations in the digestive glands of mussels from the CW and NCW tanks shows that the highest value (1700 mg/kg d.w.) was found in the CW mussels collected after 13 days. In NCW, the mean aluminum concentration in digestive glands during the test was 281 mg/kg d.w. The rapid concentration decrease in digestive glands is probably due to the inhibition of filtering activity due to valve closure at the high concentration as well as the induction of the detoxification response.

Mussel farming as a nutrient reduction measure in the Baltic Sea: consideration of nutrient biogeochemical cycles.

Nutrient loads from the land to the sea must be reduced to combat coastal eutrophication. It has been suggested that further mitigation efforts are needed in the brackish Baltic Sea to decrease nutrients, especially in eutrophic coastal areas. Mussel farming is a potential measure to remove nutrients directly from the sea. Mussels consume phytoplankton containing nitrogen (N) and phosphorus (P); when the mussels are harvested these nutrients are removed from the aquatic system. However, sedimentation of organic material in faeces and pseudo-faeces below a mussel farm consumes oxygen and can lead to hypoxic or even anoxic sediments causing an increased sediment release of ammonium and phosphate. Moreover, N losses from denitrification can be reduced due to low oxygen and reduced numbers of bioturbating organisms. To reveal if mussel farming is a cost-effective mitigation measure in the Baltic Sea the potential for enhanced sediment nutrient release must be assessed.

Integrated biomarker assessment of the effects exerted by treated produced water from an onshore natural gas processing plant in the North Sea on the mussel Mytilus edulis.

The biological impact of a treated produced water (PW) was investigated under controlled laboratory conditions in the blue mussel, Mytilus edulis. Mussel health status was assessed using an integrated biomarker approach in combination with chemical analysis of both water (with SPMDs), and mussel tissues. Acyl-CoA oxidase activity, neutral lipid accumulation, catalase activity, micronuclei formation, lysosomal membrane stability in digestive cells and haemocytes, cell-type composition in digestive gland epithelium, and the integrity of the digestive gland tissue were measured after 5 week exposure to 0%, 0.01%, 0.1%, 0.5% and 1% PW. The suite of biomarkers employed were sensitive to treated PW exposure with significant sublethal responses found at 0.01-0.5% PW, even though individual chemical compounds of PW were at extremely low concentrations in both water and mussel tissues. The study highlights the benefits of an integrated biomarker approach for determining the potential effects of exposure to complex mixtures at low concentrations. Biomarkers were integrated in the Integrative Biological Response (IBR/n) index.

A historical assessment of coastal contamination in Birch Harbor, Maine based on the analysis of mussels collected in the 1940s and the Mussel Watch Program.

Coastal contamination in the 1940s was assessed based on analysis of canned blue mussels presumably collected from Birch Harbor, Maine, USA. Analytical results on legacy organic contaminants were compared to long-term National Oceanic and Atmospheric Administration (NOAA) Mussel Watch (MW) monitoring data to estimate the degree of coastal contamination before World War II (WWII) when many synthetic organic compounds were first introduced into the environment. While dieldrin and chlordane were not detected in the canned mussels, dichloro-diphenyl-trichloroethane (DDT) and hexachlorocyclohexanes (HCHs) were present at lower concentrations relative to the more recent MW data. Polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) were detected, and the later were significantly higher in canned mussels relative to the MW data (p<0.05). Furthermore, moving average analysis applied to the MW data depicted three-phased temporal trend patterns (increase-decrease-steady state) for virtually all contaminants indicating an overall increased coastal contamination in post WWII era.

An assessment of persistent organic pollutants (POPs) in wild and rope grown blue mussels (Mytilius edulis) from Scottish coastal waters.

Farmed, rope grown mussels (Loch Etive and Loch Ewe, both on the west coast of Scotland) and wild mussels (Straad on the west coast and Shell Bay and Aberdeen Breakwater, both on the east coast of Scotland) were collected on a monthly basis and analysed for persistent organic pollutants (POPs) with the aim of assessing the status of Scottish mussels, with respect to concentrations of POPs, and investigating site-specific and seasonal differences. Samples were analysed for polycyclic aromatic hydrocarbons (PAHs), polybrominated diphenyl ethers (PBDEs) and chlorobiphenyls (CBs). Total PAH (2- to 6-ring parent and alkylated) concentrations in mussels from three pristine sites (ref. 1: ICES Marine Chemistry Working Group Report 2008, http://www.ices.dk/reports/MHC/2007/MCWG07.pdf) (Loch Etive, Loch Ewe and Straad) were significantly lower than in mussels from sites with greater coastal influences (Aberdeen Breakwater and Shell Bay). Seasonal trends in the PAH concentrations were evident at the pristine sites, with concentrations being significantly higher for samples collected between November and March compared to those collected between April and October. The PAH data was assessed using a recently proposed traffic light system, based on the assessment criteria adopted by OSPAR for use in the 2008 Coordinated Environmental Monitoring Programme (CEMP) assessment. Concentrations were compared to Background Assessment Concentrations (BAC; blue/green transition) and Environmental Assessment Concentrations (EACs; green/red transition). All sites were classed as 'green' for the PAHs analysed, being below EACs, where available. The pristine sites were also below BACs for some PAHs and therefore would be classed as 'blue' for these PAHs. CBs and PBDEs were measured in mussels collected between 2006 and 2008 inclusive. Concentrations for CB and PBDEs were significantly higher in the Aberdeen Breakwater mussels than for all other sites. Concentrations at all sites were low with many congeners being below the detection limits for both contaminant groups. No seasonal trends were observed in the CBs or PBDE concentrations or composition at any of the five sites. The green-red transition (described by the EACpassive) was exceeded for CB118 in mussels from Aberdeen Breakwater and surprisingly at the pristine site of Loch Etive. As such both sites were classed as 'red'. All other ICES7 CBs were below EACs and therefore classed as 'blue' or 'green'.

Mimicking biopolymers on a molecular scale: nano(bio)technology based on engineered proteins.

Proteins are ubiquitous biopolymers that adopt distinct three-dimensional structures and fulfil a multitude of elementary functions in organisms. Recent systematic studies in molecular biology and biotechnology have improved the understanding of basic functional and architectural principles of proteins, making them attractive candidates as concept generators for technological development in material science, particularly in biomedicine and nano(bio)technology. This paper highlights the potential of molecular biomimetics in mimicking high-performance proteins and provides concepts for applications in four case studies, i.e. spider silk, antifreeze proteins, blue mussel adhesive proteins and viral ion channels.

Modeling the effect of temperature on bioaccumulation of metals by a marine bioindicator organism, Mytilus edulis.

To determine whether regional or seasonal variability in water temperatures might affect the bioaccumulation of metals by marine invertebrates, we used a biokinetic/bioenergetic approach to model metal bioaccumulation (Ag, Am, Cd, Co, Se, and Zn) from dietary and dissolved sources by blue mussels, Mytilus edulis, acclimated at 2 and 12 degrees C. Accumulation of metal from the aqueous phase was not affected by temperature. However, Ag, Am, and Zn from diet were respectively accumulated up to 5.7-fold, 5.3-fold, and 2-fold more effectively at 2 degrees C than at 12 degrees C, largely because these three metals were assimilated from food more effectively at the lower temperature. In contrast, bioaccumulation of Cd, Co, Se from diet was not substantially affected by temperature even though efflux constants for these metals were up to 4-fold lower at 2 degrees C than at 12 degrees C. Total bioaccumulation of Ag, Am, and Zn was up to 1.7-3.6-fold higher at 2 degrees C than at 12 degrees C, with the largest differences predicted for high food conditions. Temperature-related variability in bioaccumulation of metals should be considered when interpreting patterns in metal tissue concentrations and when adapting management strategies developed for temperate seas to polar areas.

Characterization of fatty acid esters of okadaic acid and related toxins in blue mussels (Mytilus edulis) from Norway.

Marine algal toxins of the okadaic acid group can occur as fatty acid esters in blue mussels, and are commonly determined indirectly by transformation to their parent toxins by alkaline hydrolysis. Some data are available regarding the identity of the fatty acid esters, mainly of palmitic acid (16:0) derivatives of okadaic acid (OA), dinophysistoxin-1 (DTX1) and dinophysistoxin-2 (DTX2). Other fatty acid derivatives have been described, but with limited mass spectral data. In this paper, the mass spectral characterization of the [M-H](-) and [M+Na](+) ions of 16 fatty acid derivatives of each of OA, DTX1 and DTX2 is presented. The characteristic fragmentation of [M+Na](+) ions of OA analogues provided a useful tool for identifying these, and has not been described previously. In addition, a set of negative ion multiple reaction monitoring (MRM) methods was developed for direct determination of 16 fatty acid esters of OA, 16 fatty acid esters of DTX1 and 16 fatty acid esters of DTX2 in shellfish extracts. The MRM methods were employed to study the profiles of fatty acid esters of OA analogues in blue mussels and to compare these with fatty acid ester profiles reported for other groups of marine algal toxins.

A model for estimating the potential biomagnification of chemicals in a generic food web: preliminary development.

GOAL, SCOPE AND BACKGROUND: Bioaccumulation and biomagnification of organic pollutants have been increasingly assessed and modeled during the last years. Due to the complexity of these processes and the large variability of food webs, setting generic assessments for these parameters is really difficult. Equilibrium models, based on a compound's lipophylicity, are the main tool in regulatory proposals, such as for identifying Persistent, Bioaccumulative and Toxic Substances (PBTs), although a refinement has been claimed by the scientific community. Toxicokinetic studies offer an alternative for these estimations, where biomagnification is modeled as a succession of bioaccumulation processes, each one regulated by toxicokinetic parameters. METHODS: A review of kinetic models covering species belonging to different trophic levels and with different ecological behavior has been conducted. The results were employed for setting a conceptual model for estimating the biomagnification potential in a generic food web, which was mathematically implemented through system dynamic models developed under data sheet software. Crystal Ball was then employed for allowing Monte Carlo based probabilistic calculations. Bioaccumulation laboratory assays have been performed to estimate toxicokinetic parameters in mussels (Mytilus edulis) with two PAHs (chrysene and benzo[a]pyrene). The contamination was delivered via food. The exposure period lasted more than one month followed then by a depuration phase. The contaminant content was determined on an individual basis on five replicates. RESULTS AND DISCUSSION: . The reviewed information suggested the development of a tiered conceptual biomagnification model, starting with a simplified food chain which can be refined to more realistic and complex models in successive levels. CONCLUSIONS: The mathematical implementation of the conceptual model offers tools for estimating the potential for bioaccumulation and biomagnification of chemicals under very different conditions. The versatility of the model can be used for both comparative estimations and for validating the model. RECOMMENDATIONS AND PERSPECTIVES: Since bioaccumulation and biomagnification processes are crucial elements for a proper risk assessment of chemicals, their estimation by mathematical models has been widely tested. However, inregulatory assessments, too simplistic models are still being used quite often. The biomagnification model presented in this study should be amore accurate alternative to these models. In comparison to other previously published biomagnification models, the present one covers the time variation of bioaccumulation using just a few toxicokinetic parameters.

Assessment of the quantitative determination of paralytic shellfish poisoning toxins by pre-column derivatization and elimination of interfering compounds by solid-phase extraction.

Monitoring programmes for paralytic shellfish poisoning toxins in bivalve molluscs still rely heavily on the use of mouse bioassays (MBA) for consumer protection. A high-performance liquid chromatography (HPLC) methodology (Lawrence method) was implemented in 1996 in the Portuguese monitoring programme as a complementary means of analysis. Comparison between MBA and HPLC was done at the time only by a qualitative approach due to the scarce number of positive samples tested. More quantitative data were obtained recently when studying toxin profiles in Moroccan shellfish, and the correlation found between these two methodologies is reported here for the first time. Two different matrices were studied: blue mussel and the giant cockle Acanthocardia tuberculatum. A good linear correlation was obtained for both matrices. However, a second-degree polynomial best fitted the data at both low and high extremes of toxicity. According to the HPLC quantitative results, 13% of false-negatives could be obtained by MBA due to an underestimation of toxicity near the limit of detection of the MBA. Difficulties on relying solely on HPLC for consumer protection have been aroused with uncommon matrices, such as imported clams or crustaceans, due to the presence of high concentrations of interfering compounds. The solid-phase extraction step of the Lawrence method was implemented to eliminate an unknown compound that could be mistaken for saxitoxin, and an 80% reduction of another common unknown compound eluting close to decarbamoylsaxitoxin. The implementation of the HPLC methodology achieved so far allows a high degree of consumer protection without the need to resource to animal sacrifice.