Exceptions and exemptions under the ballast water management convention - Sustainable alternatives for ballast water management?

The International Convention for the Control and Management of Ships' Ballast Water and Sediments (BWM Convention) aims to mitigate the introduction risk of harmful aquatic organisms and pathogens (HAOP) via ships' ballast water and sediments. The BWM Convention has set regulations for ships to utilise exceptions and exemptions from ballast water management under specific circumstances. This study evaluated local and regional case studies to provide clarity for situations, where ships could be excepted or exempted from ballast water management without risking recipient locations to new introductions of HAOP. Ships may be excepted from ballast water management if all ballasting operations are conducted in the same location (Regulation A-3.5 of the BWM Convention). The same location case study determined whether the entire Vuosaari harbour (Helsinki, Finland) should be considered as the same location based on salinity and composition of HAOP between the two harbour terminals. The Vuosaari harbour case study revealed mismatching occurrences of HAOP between the harbour terminals, supporting the recommendation that exceptions based on the same location concept should be limited to the smallest feasible areas within a harbour. The other case studies evaluated whether ballast water exemptions could be granted for ships using two existing risk assessment (RA) methods (Joint Harmonised Procedure [JHP] and Same Risk Area [SRA]), consistent with Regulation A-4 of the BWM Convention. The JHP method compares salinity and presence of target species (TS) between donor and recipient ports to indicate the introduction risk (high or low) attributed to transferring unmanaged ballast water. The SRA method uses a biophysical model to determine whether HAOP could naturally disperse between ports, regardless of their transportation in ballast water. The results of the JHP case study for the Baltic Sea and North-East Atlantic Ocean determined that over 97% of shipping routes within these regions resulted in a high-risk indication. The one route assessed in the Gulf of Maine, North America also resulted in a high-risk outcome. The SRA assessment resulted in an overall weak connectivity between all ports assessed within the Gulf of the St. Lawrence, indicating that a SRA-based exemption would not be appropriate for the entire study area. In summary, exceptions and exemptions should not be considered as common alternatives for ballast water management. The availability of recent and detailed species occurrence data was considered the most important factor to conduct a successful and reliable RA. SRA models should include biological factors that influence larval dispersal and recruitment potential (e.g., pelagic larval duration, settlement period) to provide a more realistic estimation of natural dispersal.

Temporal Trends (1981-2013) of Per- and Polyfluoroalkyl Substances and Total Fluorine in Baltic cod (Gadus morhua).

Temporal trends from 1981 to 2013 of 28 per- and polyfluoroalkyl substances (PFASs) were investigated in liver tissue of cod (Gadus morhua) sampled near southeast Gotland, in the Baltic Sea. A total of 10 PFASs were detected, with ∑28 PFAS geometric mean concentrations ranging from 6.03 to 23.9 ng/g ww. Perfluorooctane sulfonate (PFOS) was the predominant PFAS, which increased at a rate of 3.4% per year. Most long-chain perfluoroalkyl carboxylic acids increased at rates of 3.9 to 7.3% per year except for perfluorooctanoate (PFOA), which did not change significantly over time. The perfluoroalkyl acid precursors perfluorooctane sulfonamide (FOSA) and 6:2 fluorotelomer sulfonic acid were detected, of which the former (FOSA) declined at a rate of -4.4% per year, possibly reflecting its phase-out starting in 2000. An alternate time trend analysis from 2000 to 2013 produced slightly different results, with most compounds increasing at slower rates compared to the entire study period. An exception was perfluorohexane sulfonate (PFHxS), increasing at a faster rate of 3.7% measured from 2000 on, compared to the 3.0% per year measured starting from 1981. Analysis of the total fluorine content of the samples revealed large amounts of unidentified fluorine; however, its composition (organic or inorganic) remains unclear. Significant negative correlations were found between concentrations of individual PFASs (with the exception of PFOS) and liver somatic index. In addition, body length was negatively correlated with PFOA and perfluorononanoate, but positively correlated with perfluorododecanoate (PFDoDA) and FOSA. Additional studies on endocrine, immunological, and metabolic effects of PFAS in marine fish are essential to assess the environmental risk of these substances. Environ Toxicol Chem 2020;39:300-309. © 2019 SETAC.

Toxic effects of chemical warfare agent mixtures on the mussel Mytilus trossulus in the Baltic Sea: A laboratory exposure study.

Baltic blue mussels (Mytilus trossulus) were implemented to assess potential toxicity, health impairments and bioaccumulation of dumped chemical warfare agents on marine benthic organisms. Mussels were collected from a pristine cultivation side and exposed under laboratory conditions to different mixtures of chemical warfare agents (CWAs) related phenyl arsenic compounds, Clark I and Adamsite as well as chloroacetophenone. Using a multi-biomarker approach, mussels were assessed thereafter for effects at different organisational levels ranging from geno-to cytotoxic effects, differences in enzyme kinetics and immunological responses. In an integrated approach, chemical analysis of water and tissue of the test organisms was performed in parallel. The results show clearly that exposed mussels bioaccumulate the oxidized forms of chemical warfare agents Clark I, Adamsite (DAox and DMox) and, to a certain extent, also chloroacetophenone into their tissues. Adverse effects in the test organisms at subcellular and functional level, including cytotoxic, immunotoxic and oxidative stress effects were visible. These acute effects occurred even at the lowest test concentration.

Integrated monitoring of chemicals and their effects on four sentinel species, Limanda limanda, Platichthys flesus, Nucella lapillus and Mytilus sp., in Seine Bay: A key step towards applying biological effects to monitoring.

The International workshop on Integrated Assessment of CONtaminants impacts on the North sea (ICON) provided a framework to validate the application of chemical and biological assessment thresholds (BACs and EACs) in the Seine Bay in France. Bioassays (oyster larval anomalies, Corophium arenarium toxicity assay and DR Calux) for sediment and biomarkers: ethoxyresorufin-O-deethylase (EROD) activity, acetylcholinesterase (AChE) activity, lysosomal membrane stability (LMS), DNA strand breaks using the Comet assay, DNA adducts, micronucleus (MN), PAH metabolites, imposex, intersex and fish external pathologies were analysed in four marine sentinel species (Platichthys flesus, Limanda limanda, Mytilus sp. and Nucella lapilus). Polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and heavy metals were analysed in biota and sediment. Results for sediment and four species in 2008-2009 made it possible to quantify the impact of contaminants using thresholds (Environmental Assessment Criteria/EAC2008: 70% and EAC2009: 60%) and effects (EAC2008: 50% and EAC2009: 40%) in the Seine estuary. The Seine estuary is ranked among Europe's most highly polluted sites.

Immunomodulating effects of environmentally realistic copper concentrations in Mytilus edulis adapted to naturally low salinities.

The monitoring of organisms' health conditions by the assessment of their immunocompetence may serve as an important criterion for the achievement of the Good Environmental Status (GES) as defined in the Marine Strategy Framework Directive (EU). In this context, the complex role of natural environmental stressors, e.g. salinity, and interfering or superimposing effects of anthropogenic chemicals, should be carefully considered, especially in scenarios of low to moderate contamination. Organisms from the Baltic Sea have adapted to the ambient salinity regime, however energetically costly osmoregulating processes may have an impact on the capability to respond to additional stress such as contamination. The assessment of multiple stressors, encompassing natural and anthropogenic factors, influencing an organisms' health was the main aim of the present study. Immune responses of Mytilus edulis, collected and kept at natural salinities of 12‰ (LS) and 20‰ (MS), respectively, were compared after short-term exposure (1, 7 and 13 days) to low copper concentrations (5, 9 and 16 µg/L Cu). A significant interaction of salinity and copper exposure was observed in copper accumulation. LS mussels accumulated markedly more copper than MS mussels. No combined effects were detected in cellular responses. Bacterial clearance was mostly achieved by phagocytosis, as revealed by a strong positive correlation between bacterial counts and phagocytic activity, which was particularly pronounced in LS mussels. MS mussels, on the other hand, seemingly accomplished bacterial clearance by employing additional humoral factors (16 µg/L Cu). The greatest separating factor in the PCA biplot between LS and MS mussels was the proportion of granulocytes and hyalinocytes while functional parameters (phagocytic activity and bacterial clearance) were hardly affected by salinity, but rather by copper exposure. In conclusion, immune responses of the blue mussel may be suitable and sensitive biomarkers for the assessment of ecosystem health in brackish waters (10-20‰S).

The activity of macrophage aggregates in the liver of flounder (Platichthys flesus) and wrasse (Symphodus melops) is associated with tissue damage.

The phagocytic activity of macrophage aggregates (MAs) in the liver of fish is characterized by high acid phosphatase activity and generation of reactive oxygen species (ROS). Question of the present study was whether these activities were associated with damage of adjacent and surrounding liver cells and the macrophages itself. Thus, the lysosomal membrane stabilities (LMS) of two different populations of lysosomes (LMS1, short destabilization periods, and LMS2, longer destabilization periods) of liver cells close to MAs of different phagocytic activity were measured as indicator for cellular integrity by computer assisted image analysis. They were then compared to the LMS of hepatocytes remote from any MA. In addition, LMS was also assessed inside the MAs. Studies were performed on adult European flounder (Platichthys flesus) and corkwing wrasse (Symphodus melops) caught at clean and polyaromatic hydrocarbon (PAH)-contaminated field locations of the German Bight and the Karmoy peninsula at the West coast of Norway. In the hepatocytes adjacent to MAs which showed medium to high phagocytic activity, LMS was significantly decreased with a reduction of both, first (LMS1), and second (LMS2) destabilization periods in both species. LMS1 inside of these MAs was extremely low, whereas LMS2 was sometimes even higher compared to the liver cells. We suggest that LMS1 represents phagosomes which had contact to ROS and LMS2 represents less affected primary lysosomes. In single flounder and wrasse with liver tumors, MAs in the vicinity of these tumors showed low phagocytic activity and were not associated with adverse effects on adjacent tumor cells. Inside these MAs, LMS1 was higher compared to MAs with high phagocytic activity in wrasse from the reference site, indicating that cytotoxicity was not involved in their phagocytic deficiency.

Effects of nanoparticles in Mytilus edulis gills and hepatopancreas - a new threat to marine life?

Every day new extraordinary properties of nanoparticles (a billionth of a meter) are discovered and worldwide millions are invested into nanotechnology and nanomaterials. Risks to marine organisms are still not fully understood and biomarkers to detect health effects are not implemented, yet. We used the filter feeding blue mussel as a model to analyse uptake and effects of nanoparticles from glass wool, a new absorbent material suggested for use in floating oil spill barriers. In both, gills and hepatopancreas we analysed uptake of nanomaterials by transmission electronmicroscopy (TEM) in unstained ultrathin sections over a period of up to 16 days. Lysosomal stability and lipofuscin content as general indicators of cellular pathology and oxidative stress were also measured. As portals of uptake, diffusion and endocytosis were identified resulting in nanoparticle accumulation in endocytotic vesicles, lysosomes, mitochondria and nuclei. Dramatic decrease of lysosomal membrane stability occurred after 12h of exposure. Lysosomal damage was followed by excessive lipofuscin accumulation indicative of severe oxidative stress. Increased phagocytosis by granulocytes, autophagy and finally apoptosis of epithelial cells of gills and primary and secondary digestive tubules epithelial cells indicated progressive cell death. These pathological responses are regarded as general indices of toxic cell injury and oxidative stress. By the combinational use of biomakers with the ultrastructural localisation of nanoparticle deposition, final evidence of cause-effect relationships is delivered.

The liver of wrasse - morphology and function as a mirror of point source chemical impact.

Corkwing wrasse (Symphodus melops L.), a protogynous, non-migratory lipfish species, living close to rocky shores was chosen as an indicator species for the monitoring of biological effects of contaminants. Fish were caught by local fisherman at the Norwegian west coast in fjord sites within the framework of the EU BEEP project. The sites represented different point source impacts of (I) copper (a former copper mine), (II) polycyclic aromatic hydrocarbons (PAHs, aluminium smelter discharge), (III) formaldehyde plus PAHs (kelp-factory and influence of the aluminium smelter). Livers of wrasse were studied for histopathological alterations and compared to healthy livers of fish from a reference location. Besides liver morphology, different functional and metabolic parameters were measured to link pathological alterations to functional disorders. The integrity of the lysosomal compartment was tested by the assessment of lysosomal membrane stability (lys), and the accumulation of neutral lipids and lipofuscin. Activity and intracellular localisation of the NADPH-producing enzymes in the liver were assessed histochemically and measured by computer-assisted image analysis. Histopathological alterations were most severe at the site impacted by formaldehyde and PAHs. These findings were associated with highest tumor prevalence, lowest membrane stabilities in hepatocytes and highest accumulation rates of lipofuscin in the liver. The activities of the NADPH-producing enzymes phosphogluconate dehydrogenase (PGDH) and glucose-6-phosphate dehydrogenase (G6PDH) were significantly lower compared to unimpacted reference fish. Histopathological alterations showed clear differences dependent on the input source. Potential links between specific contaminant impact and functional and morphological disorders are discussed.

Indices for the assessment of environmental pollution of the Baltic Sea coasts: integrated assessment of a multi-biomarker approach.

Two mathematical methods to assess the "health status" of flounder (Platichthys flesus), eelpout (Zoarces viviparus) and blue mussel (Mytilus spp.) populations of the Baltic Sea were applied on selected biomarker data collected during the EU project "BEEP" (Biological Effects of Environmental Pollution on Marine Coastal Ecosystems). The Bioeffect Assessment Index (BAI) and the Integrated Biomarker Index (IBR) combine different biomarkers to single values, which can be used to describe the toxically-induced stress level of populations in different areas. Both indices determined here produced essentially similar results, which in most cases agreed with the known contamination levels in the different study areas. Advantages and limitations of index applications and interpretations are critically discussed. The use of indices provides comprehensive information about biological effects of pollution in marine organisms and may therefore serve as a useful tool for environmental management by ranking the pollution status of marine coastal areas.

Measurements of biomarker levels in flounder (Platichthys flesus) and blue mussel (Mytilus trossulus) from the Gulf of Gdansk (southern Baltic).

In the framework of the EU funded BEEP project a set of biomarkers, gross morphometric indices and tissue concentrations of selected organic pollutants were measured in flounder (Platichthys flesus) and mussels (Mytilus trossulus) collected twice a year (April and October) from three sites in the inner Gulf of Gdansk between autumn 2001 and spring 2003. In flounder, seasonal differences in most biomarkers were observed, but no correlations with tissue pollutant levels could be found. In mussels, highly variable levels in biomarker responses were seen, but no clear seasonal or spatial trends, directly related to tissue concentrations, could be established. The observed biomarkers distribution the study sites are probably mostly caused by interannual, seasonal and individual variability and, in case of flounder, possibly by exchange of stocks between the sampling sites.

Biomarker responses in flounder (Platichthys flesus) and mussel (Mytilus edulis) in the Klaipeda-Butinge area (Baltic Sea).

During the EU project BEEP a battery of biomarkers was applied in flounder (Platichthys flesus) and the blue mussel (Mytilus edulis) collected at three locations off the Lithuanian coast (Baltic Sea) in June and September 2001 and 2002. The elevated biomarker responses in specimens sampled in September 2001 were apparently related to the extensive dredging activities in the Klaipeda port area and subsequent dumping of contaminated sediments. High concentrations of organic pollutants (organochlorines and PBDEs) were also measured in the tissues of both indicator species. In addition, response levels of genotoxicity, cytotoxicity, immunotoxicity as well as concentrations of PAH metabolites in the bile of flounder showed elevations in 2002 after an oil spill in the Butinge oil terminal in November 2001. In flounder, biomarker measurements 10 months after the spill indicated recovery processes but in mussels a high level of genotoxicity could still be observed 22 months later. The present study illustrates the usefulness of the multi-biomarker approach in the detection of biological effects of pollution in this region of the Baltic Sea.

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.

Liver histopathology in Baltic flounder (Platichthys flesus) as indicator of biological effects of contaminants.

Results are presented of a study on liver histopathology in Baltic flounder (Platichthys flesus) carried out in 2001 and 2002 in four coastal sampling areas of the Baltic Sea: Kvädöfjärden (Swedish east coast, reference area), Klaipeda-Butinge (Lithuanian coast), Gulf of Gdansk (Polish coast), and Wismar Bay (German coast) within the framework of the EU-funded BEEP project. Liver lesions were diagnosed and categorised using standardised methodologies and, for a spatial and temporal assessment of the prevalence and types of lesions detected, a scoring system was applied, involving the calculation of mean histopathology lesion scores. 83.0% of the 436 female flounder examined (size range: 20-43 cm total length, age range: 2-8 years) were affected by liver lesions, out of which 74.3% were assigned to the category of non-specific, 3.4% to the category of early toxicopathic non-neoplastic, 4.6% to the category of pre-neoplastic and 0.7% to the category of neoplastic lesions. Mean lesions scores were highest in the areas at the Lithuanian and Swedish coast and there is indication of an impact of the age structure of the flounder populations studied, the sampling season as well as of contaminant effects.

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.

Effects of dredging in Göteborg harbor, Sweden, assessed by biomarkers in eelpout (Zoarces viviparus).

We used a battery of biomarkers in fish to study the effects of the extensive dredging in Göteborg harbor situated at the river Göta alv estuary, Sweden. Eelpout (Zoarces viviparus) were sampled along a gradient into Göteborg harbor, both before and during the dredging. Biomarker responses in the eelpout before the dredging already indicated that fish in Göteborg harbor are chronically affected by pollutants under normal conditions compared to those in a reference area. However, the results during the dredging activities clearly show that fish were even more affected by remobilized pollutants. Elevated ethoxyresorufin-O-deethylase activities and cytochrome P4501A levels indicated exposure to polycyclic aromatic hydrocarbons. Elevated metallothionein gene expression indicated an increase in metal exposure. An increase in general cell toxicity, measured as a decrease in lysosomal membrane stability, as well as effects on the immune system also could be observed in eelpout sampled during the dredging. The results also suggest that dredging activities in the Göta alv estuary can affect larger parts of the Swedish western coast than originally anticipated. The present study demonstrates that the application of a set of biomarkers is a useful approach in monitoring the impact of anthropogenic activities on aquatic environments.