Benthic biodiversity indices versus salinity gradient in the southern Baltic Sea.

The need to assess the environmental status of marine and coastal waters according to the EU Water Framework Directive (WFD) encouraged the design of specific biotic indices to evaluate the response of benthic communities to human-induced changes in water quality. In the present study three of these indices, the traditional Shannon Wiener Index (H') and the more recently published AMBI (AZTI' Marine Biotic Index) and BQI (Benthic Quality Index), were tested along a salinity gradient in the southern Baltic Sea. The comparison of the three indices demonstrates that in the southern Baltic Sea the ecological quality (EcoQ) classification based on macrozoobenthic communities as indicator greatly depends on the biotic index chosen. We found a significant positive relation between species number, H', BQI and salinity resulting in EcoQ status of "Bad", "Poor" or "Moderate" in areas with a salinity value below 10 psu. The AMBI was less dependent on salinity but appear to partly overestimate the EcoQ status. Presently none of these biotic indices appear to be adjusted for application in a gradient system as given in the southern Baltic Sea. A potential approach describing how to overcome this limitation is discussed.

Interactions between climate change and contaminants.

There is now general consensus that climate change is a global threat and a challenge for the 21st century. More and more information is available demonstrating how increased temperature may affect aquatic ecosystems and living resources or how increased water levels may impact coastal zones and their management. Many ecosystems are also affected by human releases of contaminants, for example from land based sources or the atmosphere, which also may cause severe effects. So far these two important stresses on ecosystems have mainly been discussed independently. The present paper is intended to increase awareness among scientists, coastal zone managers and decision makers that climate change will affect contaminant exposure and toxic effects and that both forms of stress will impact aquatic ecosystems and biota. Based on examples from different ecosystems, we discuss risks anticipated from contaminants in a rapidly changing environment and the research required to understand and predict how on-going and future climate change may alter risks from chemical pollution.

Protein responses in blue mussels (Mytilus edulis) exposed to organic pollutants: a combined CYP-antibody/proteomic approach.

Polyclonal antibodies were raised against highly conserved, trans-metazoan sequences of cytochrome P450 (CYP) families 2 and 4 and used to investigate responses in the common blue mussel (Mytilus edulis) exposed to various organic contaminants. The results were evaluated by means of cross-reacting proteins on Western blots of both one- and two-dimensional electrophoresis gels, and by scanning spectroscopy measurements of total CYP content. Furthermore, a proteomic approach was applied aimed at elucidating exposure-related protein changes in a more general term. Identities of isolated proteins were searched by means of peptide mass fingerprints obtained from MALDI-TOF MS analyses. The results demonstrated that both antibodies rendered several cross-reactive bands when probed on Western blots. The most obvious cross-reaction of the CYP2 antibody was with a strongly expressed protein of size approximately 57kDa, pI 4.5-4.6, whereas the CYP4 antibody cross-reacted with a protein of size approximately 55kDa, pI 5.6. However, expression of cross-reacting proteins did not change as a result of the exposures, and resulted only in small and insignificant fluctuations in total CYP content. As a contrast, silver-stained 2DE gels showed that several microsomal proteins were affected in individuals exposed to diallylphthalate as well as crude oil, with and without a spike of alkylphenols and PAHs. Mass spectrometry based analyses of excised, trypsin-digested spots did so far not decipher the identities of the proteins affected by the exposures, nor of those cross-reacting with CYP2 and CYP4 antibodies. This study has underlined the power of the proteomic approach in environmental toxicology, although protein identification was not successful. The missing identities of the proteins cross-reacting with the CYP2- and CYP4-antibodies does not enable a clear conclusion as to whether or not these peptides actually represent CYP iso-enzymes.

Expression of cytoskeletal proteins, cross-reacting with anti-CYP1A, in Mytilus sp. exposed to organic contaminants.

The possible use of cytoskeletal components as biomarkers of organic pollution in mussels has been investigated. Responses of non-muscular actin and tropomyosin (TM), two bivalve proteins that were recently demonstrated to cross-react with anti-fish-CYP1A, were analysed in digestive tissue of blue mussels (Mytilus sp.) exposed to a wide range of organic contaminants. The results were evaluated with ELISA and Western blot assays, utilising commercial monoclonal antibodies, and compared with expression of Hsp70, a marker of chemical stress. Furthermore, mussels were sampled from the Baltic Sea at sites with different degrees of pollution to assess the expression of these proteins, and to monitor seasonal changes in relation to energy reserves and water temperature. The results demonstrated that expression of microsomal actin was significantly higher (p<0.02) in mussels exposed to a brominated flame retardant (BDE-47), and lower, however not significantly, in specimens exposed to crude oil, alone and spiked with alkylphenols and PAHs. Hsp70 was strongly induced in all exposure groups, which also included bisphenol A and diallylphthalate. Furthermore, microsomal actin exhibited seasonal variations, and expression was negatively correlated with water temperature. No correlation was seen between actin and the microfilament-binding protein TM, indicating that regulation of these two cytoskeletal components are not coupled. Furthermore, parallel and significant (p<0.05) up-regulations of TM and Hsp70 were seen in individuals sampled from a strongly polluted field site, whereas the seasonal analysis showed that TM expression was positively correlated with energy reserves (total glycogen content) in mussels, suggesting the use of TM as a marker of growth. In conclusion, this study has demonstrated the cytoskeleton to be a target of contaminants in mussels, calling for further attention. Exposure-induced increase of microsomal actin can be interpreted either as stimulated actin synthesis, or re-arrangements of the dynamic microfilaments.

Monitoring biological effects of pollution in the Baltic Sea: neglected--but still wanted?

In the Baltic Sea, studies regarding biological effects of contaminants are relatively few, partly due to political and economical reasons, specific hydrographic characteristics, and a strong eutrophication-targeted research focus during the past three decades. The development of a monitoring strategy concerning biological effects and its implementation into environmental monitoring programmes in the Baltic Sea is lagging behind the progress currently taking place in most of western and southern Europe. The pan-European project BEEP (Biological Effects of Environmental Pollution on Marine Coastal Ecosystems, 2001-2004) included the Baltic Sea as one of the target areas for the evaluation of a suite of biological effects indicators in European coastal waters. The main aims of the BEEP project are described and how the expected outcome for the Baltic Sea could provide the needed "baseline" information and expertise for a biological effect monitoring and contribute to harmonise environmental monitoring programmes within the EU.

Cytogenetic and cytotoxic effects in gill cells of the blue mussel Mytilus spp. from different zones of the Baltic Sea.

Frequency of micronuclei (MN) and other nuclear abnormalities (nuclear buds, bi-nucleated and fragmented-apoptotic cells) was analysed in gill cells of the blue mussel (Mytilus spp.) from selected coastal sites in the Baltic Sea--Kvädöfjärden (Sweden), Klaipeda-Butinge zone (Lithuania), Gulf of Gdansk (Poland) and Wismar Bay (Germany). Samples were collected from 650 specimens during bi-annual sampling campaigns in 2001 and 2002. The lowest frequency of MN (0.37 MN/1000 cells) was found in blue mussels from the reference site (Kvädöfjärden). The highest MN values (up to 6.7 MN/1000 cells) were registered in blue mussels from the Gulf of Gdansk in autumn 2001 and 2002, and at Wismar Bay in spring 2001 (up to 5.06 MN/1000 cells). Gradients of MN incidences were observed when comparing the three studied locations in Wismar Bay, and at the Lithuanian coast before the crude oil spill in the Butinge oil terminal. Moreover, significant seasonal and inter-location differences in the responses were documented (P<0.0001). Nuclear abnormalities were observed most frequently in blue mussels from the Gulf of Gdansk.

Biomarker responses as indication of contaminant effects in blue mussel (Mytilus edulis) and female eelpout (Zoarces viviparus) from the southwestern Baltic Sea.

During a field study performed in spring and autumn 2001 and 2002, blue mussels (Mytilus edulis) and female eelpout (Zoarces viviparus) were collected at three locations in the Wismar Bay (Baltic Sea), and several biomarkers of contaminant effects were analysed. Besides seasonal and inter-annual variations, biomarker signals were most pronounced at the location closest to Wismar Harbour (Wendorf) in both species. Lysosomal membrane stability (LMS) was lowest and acetylcholinesterase activity (AChE) was significantly reduced. Frequency of micronuclei (MN) was significantly higher (in blue mussels), indicating mutagenic effects. In eelpout elevated levels of DNA adducts, EROD induction and PAH-metabolites were measured. Metallothionein (MT), biomarker for trace metal exposure, showed a gradient only in spring. Organochlorine contaminant analyses (PCBs, DDTs) corresponded to the observed biomarker levels. The results obtained clearly demonstrate pollution effects in the southwestern Baltic Sea. Moreover, they show that a multibiomarker approach is also applicable in a brackish water environment.

Use of biliary PAH metabolites as a biomarker of pollution in fish from the Baltic Sea.

During field campaigns of the BEEP project (Biological Effects of Environmental Pollution in Marine Coastal Ecosystems) in 2001-2002, metabolites of polycyclic aromatic hydrocarbons (PAHs) were determined in bile samples from three fish species, flounder (Platichthys flesus), perch (Perca fluviatilis) and eelpout (Zoarces viviparus), from four separate areas in the Baltic Sea. Two determination methods were applied: fixed wavelength fluorescence (FF) for pyrene-type metabolites and high-performance liquid chromatography with fluorescence detection (HPLC). There was a good correlation between the FF method and 1-OH pyrene determined by HPLC. Normalisation of the FF data for absorbance at 380 nm or bile protein concentrations greatly increased variance in one third and decreased it in two thirds of the cases and resulted in a loss of significant differences (protein normalisation) between the sampling stations, but normalisation of the HPLC data had little effect on the results. The biliary PAH metabolite content was usually higher in males than in females. In perch and eelpout the biliary PAH contents were at similar levels, whereas in flounder the levels were lower. The sampling areas arranged in decreasing order of biliary PAH contents were: Wismar Bay > Gulf of Gdansk > Lithuanian coast > Kvadofjärden (reference area). It is concluded that FF with un-normalised data is a reliable and simple method for monitoring purposes and only one sex of a selected species should be used.

The BEEP project in the Baltic Sea: overview of results and outline for a regional biological effects monitoring strategy.

Field studies in the framework of the EU funded BEEP project (Biological Effects of Environmental Pollution in Marine Coastal Ecosystems, 2001-2004) aimed at validating and intercalibrating a battery of biomarkers of contaminant exposure and effects in selected indicator species in the Mediterranean, the North Atlantic and the Baltic Seas. Major strategic goals of the BEEP project were the development of a sensitive and cost-efficient biological effects monitoring approach, delivery of information and advice to end-user groups, and the implementation of a network of biomarker researchers around Europe. Based on the main results obtained in the Baltic Sea component of the BEEP the present paper summarises and assesses the applicability of biomarkers for different regions and species in this sea area. Moreover, a general strategy and some practical considerations for the monitoring of biological effects in the Baltic Sea are outlined.

Biochemical biomarkers in adult female perch (Perca fluviatilis) in a chronically polluted gradient in the Stockholm recipient (Sweden).

A battery of biochemical biomarkers and the SigmaPCB concentration in adult female perch (Perca fluviatilis) verified an aquatic pollution gradient with the city of Stockholm (Sweden) as a point source of anthropogenic substances. The investigation included both an upstream gradient, 46 km westwards through Lake Mälaren, and a downstream gradient, 84 km eastwards through the Stockholm archipelago. Besides the main gradient from Stockholm, there were strong indications of pollution coming from the Baltic Sea. The results indicated a severe pollution situation in central Stockholm, with poor health status of the perch, characterised by increased specific EROD activity in the liver, increased liver EROD somatic index, decreased AChE activity in the muscle, increased amount of DNA adducts in the liver, and a high concentration of biliary 1-pyrenol. In addition, laboratory exposure to common EROD inducers elicited an abnormal response, suggestive of chronic intoxication.

Regime shifts in the marine environment: the scientific basis and political context.

Regime shifts in the marine environment have recently received much attention. To date, however, few large-scale meta-analyses have been carried out due to insufficient data coverage and integration between sustained observational datasets because of diverse methodologies used in data collection, recording and archival. Here we review the available data on regime shifts globally, followed by a review of current and planned policies with relevance to regime shifts. We then focus on the North and Baltic Seas, providing examples of existing efforts for data integration in the MarBEF Network of Excellence. Existing gaps in data coverage are identified, and the added value from meta-analyses of multiple datasets demonstrated using examples from the MarBEF integrated data project LargeNet. We discuss whether these efforts are addressing current policy needs and close with recommendations for future integrated data networks to increase our ability to understand, identify and predict recent and future regime shifts.

Geographic and seasonal patterns and limits on the adaptive response to temperature of European Mytilus spp. and Macoma balthica populations.

Seasonal variations in seawater temperature require extensive metabolic acclimatization in cold-blooded organisms inhabiting the coastal waters of Europe. Given the energetic costs of acclimatization, differences in adaptive capacity to climatic conditions are to be expected among distinct populations of species that are distributed over a wide geographic range. We studied seasonal variations in the metabolic adjustments of two very common bivalve taxa at European scale. To this end we sampled 16 populations of Mytilus spp. and 10 Macoma balthica populations distributed from 39 degrees to 69 degrees N. The results from this large-scale comprehensive comparison demonstrated seasonal cycles in metabolic rates which were maximized during winter and springtime, and often reduced in the summer and autumn. Studying the sensitivity of metabolic rates to thermal variations, we found that a broad range of Q (10) values occurred under relatively cold conditions. As habitat temperatures increased the range of Q (10) narrowed, reaching a bottleneck in southern marginal populations during summer. For Mytilus spp., genetic-group-specific clines and limits on Q (10) values were observed at temperatures corresponding to the maximum climatic conditions these geographic populations presently experience. Such specific limitations indicate differential thermal adaptation among these divergent groups. They may explain currently observed migrations in mussel distributions and invasions. Our results provide a practical framework for the thermal ecophysiology of bivalves, the assessment of environmental changes due to climate change and its impact on (and consequences for) aquaculture.