Lysosomal membrane stability and metallothionein content in Mytilus galloprovincialis (L.), as biomarkers. Combination with trace metal concentrations

The simultaneous study of two biomarkers, the lysosomal membrane stability (LMS) of the digestive gland and the metallothionein (MT) content of the digestive gland, the gills and the mantle/gonad complex of the mussel Mytilus galloprovincialis was examined in an enclosed estuarine system in June and October 2001. Mussels were collected along the Gulf of Thermaikos (northern Greece) from stations displaying a pollution gradient, while Olympiada in the Gulf of Strymonikos was used as a reference station. In addition, the heavy metal (Cd, Pb, Cu and Zn) content, using atomic absorption spectrophotometry (AAS), were evaluated in the digestive gland, the gills and the mantle/gonad complex of mussels collected from the same sites and seasons. LMS values were significantly greater, and the MT content of the studied tissues were significantly less in mussels collected from the reference station compared to mussels from more polluted stations located in the Gulf of Thermaikos. Significant correlation was observed between the MT content of the gills and the mantle/gonad complex with the LMS values.     

Adaptation strategy of bivalve Modiolus modiolus from upwelling regions of the Kuril Islands shelf (Sea of Okhotsk) to heavy metal effects

Cd, Cu, Fe, Mn, Ni, Pb, Zn concentration in the organs (muscle, gonads, mantle, gills, digestive gland, kidney) of mussel Modiolus modiolus was studied. Mussels were collected from shelf upwelling sites of Kuril Is. Sea of Okhotsk (104 and 85 m) and, for comparison, coastal high contaminated and clean sites (Peter the Great Bay, Sea of Japan). It was found that mussels from shelf upwelling regions accumulated unusually high Cd concentration in studied organs, excepting digestive gland. Shelf mussels have evolved passive adaptation to elevated level of ambient heavy metals (distribution of heavy metal load between all organs, especially, muscle). In contrast, mussels from high contaminated sites have evolved active adaptation (strong regulation of metal concentration in the organs at the maximal permissible concentration due to increased kidney function). We supported that under natural condition, passive adaptation of aquatic organisms to increased ambient metal level have been evolved only under constant low-water temperature.     

Measurement of environmental trace-metal levels in Mediterranean coastal areas with transplanted mussels and DGT techniques

Mussels (Mytilus galloprovincialis) and diffusive gradients in thin films (DGT) devices were deployed together for three one-month periods in coastal waters of Sardinia (Italy), in order to assess the levels of Cd, Cu, Ni, and Pb in areas differently affected by anthropogenic activities. DGT devices were more sensitive than mussels in detecting differences in metal concentrations between sites, while interpretation of the biomonitoring data revealed difficulties related to the biotic and abiotic factors that can influence the measurements. Regression analysis showed a significant correlation between Cd and Pb concentrations measured in the mussel tissues and bioavailable metal levels in seawater. Moreover transplanted mussels did not accumulate Cu and Ni following pre-exposure, although DGT showed significant concentrations of bioavailable forms of these metals in seawater. The results provided an estimate of the water quality with respect to dissolved metals and pointed out the usefulness of a combination of biomonitoring and DGT techniques for a better understanding of trace metal availability in coastal waters.     

Measurement of environmental trace-metal levels in Mediterranean coastal areas with transplanted mussels and DGT techniques

Mussels (Mytilus galloprovincialis) and diffusive gradients in thin films (DGT) devices were deployed together for three one-month periods in coastal waters of Sardinia (Italy), in order to assess the levels of Cd, Cu, Ni, and Pb in areas differently affected by anthropogenic activities. DGT devices were more sensitive than mussels in detecting differences in metal concentrations between sites, while interpretation of the biomonitoring data revealed difficulties related to the biotic and abiotic factors that can influence the measurements. Regression analysis showed a significant correlation between Cd and Pb concentrations measured in the mussel tissues and bioavailable metal levels in seawater. Moreover transplanted mussels did not accumulate Cu and Ni following pre-exposure, although DGT showed significant concentrations of bioavailable forms of these metals in seawater. The results provided an estimate of the water quality with respect to dissolved metals and pointed out the usefulness of a combination of biomonitoring and DGT techniques for a better understanding of trace metal availability in coastal waters.     

Field validation, in Scotland and Iceland, of the artificial mussel for monitoring trace metals in temperate seas

The artificial mussel (AM), a novel chemical sampling device, has been developed for monitoring dissolved trace metals in marine environments. The AM consists of Chelex-100 suspended in artificial seawater within Perspex tubing and enclosed with semi-permeable polyacrylamide gel at both ends. To validate the field performance of the AM in temperate waters, we deployed AMs alongside transplanted blue mussels Mytilus edulis in coastal environments in Scotland (Holy Loch, Loch Fyne, Loch Striven and Millport) and Iceland (Reykjavikurh?fn, Gufunes, South of thornerney, Hofsvik, Hvalfj?rethur and Sandgerethi) for monitoring trace metals. While uptake patterns of Cd between the AM and M. edulis were highly comparable, discrepancies were found in the accumulation profiles of the other metals (Cu, Cr, Pb and Zn), in particular Zn. Nonetheless, the AMs gave a better resolution to accurately reveal the spatial difference in dissolved metal contamination when compared with M. edulis. AMs complement the use of mussels since AMs indicate dissolved metals in seawater, whereas uptake by mussels indicates a mixture of dissolved and particulate metals. Our results also indicated that historical metal exposure of the transplanted M. edulis could significantly confound their metal concentrations especially when the deployment period was short (i.e. <34d). This study suggested that the AM can overcome problems associated with variable biological attributes and pre-exposure history in the mussel, and provides a standardized and representative time-integrated estimate of dissolved metal concentrations in different marine environments.     

Comparison of metal accumulation between 'Artificial Mussel' and natural mussels (Mytilus galloprovincialis) in marine environments

The passive sampler called 'Artificial Mussel' (AM) developed by Wu et al. (2007) can provide a time-integrated estimate of metal concentrations in the marine environment, and offers a potential device to assess and compare metal concentration in different marine environments worldwide. The aim of this study was to compare metal accumulation on AM and natural mussel Mytilus galloprovincialis at three sites with different metal loads along the Portuguese coast for four months. M. galloprovincialis were placed in cages alongside AMs at each site. Samples were collected monthly and Cd, Cr, Cu, Pb, and Zn concentrations in whole soft tissues and AMs compared. For both Cu and Cd, the results were similar between AMs and natural mussels. Higher concentrations of Zn were observed in natural mussels, whereas the inverse was shown for Pb (about 10-fold higher). Our results showed that AMs are promising tools for assessing metal concentrations in marine environments.     

Comparative petroleum hydrocarbons levels and biochemical responses in mussels from hydrothermal vents (Bathymodiolus azoricus) and coastal environments (Mytilus galloprovincialis)

Aliphatic hydrocarbons and PAHs in the whole soft tissues of Bathymodiolus azoricus from three Mid-Atlantic Ridge hydrothermal vents (Menez-Gwen, Lucky Strike and Rainbow), and Mytilus galloprovincialis from three contaminated coastal sites in South Portugal were analysed, and its effects on the digestive gland microsomes mixed-function oxygenase system (MFO) were assessed. Aliphatic hydrocarbons levels were present in the same magnitude in both coastal and hydrothermal environments, while the UCM (unresolved complex mixture) for coastal mussels were higher than in vent mussels. In general, significantly higher PAHs concentrations were found in coastal mussels, compared to B. azoricus where low molecular weight PAHs (2-3 rings) represented the majority of PAHs contrarily to what was observed in M. galloprovincialis. The MFO components were present in both mussel species, and were detected in vent mussels for the first time. However this system seems to have different roles in species from these contrasting environments. In coastal mussels MFO responded to hydrocarbon contamination while response in hydrothermal organisms appeared to be related mainly to endogenous factors.     

Seasonal and pollution-induced variations in biomarkers of transplanted mussels within the Beagle Channel

The occurrence of biomarker variations linked to environmental factors makes it difficult to distinguish the effect of pollution. In an attempt to evaluate spatial and seasonal effects of environmental parameters on biomarker responses, mussels Mytilus edulis chilensis coming from an aquaculture farm were transplanted to several points within Ushuaia Bay (Beagle Channel) for 6 weeks in summer and winter. Activities of superoxide dismutase, catalase, glutathione-S-transferase and levels of lipid peroxidation were measured in gills and digestive gland. Cu, Zn, Fe, Cd and Pb concentrations were also assessed. Results indicated a significant effect of seasons on biological responses as well as in metal bioaccumulation showing the influence of natural factors such as dissolved oxygen, temperature and food availability. The interdependence of those environmental factors is important for the homeostasis of thermoconformers, especially regarding their oxidative metabolism and should also be taken into consideration to distinguish natural from pollution-induced variations.     

Stable isotopes and metal contamination in caged marine mussel Mytilus galloprovincialis

Metal concentrations and isotopic composition were measured in different tissues of the mussel Mytilus galloprovincialis in waters of the Balearic Islands (Western Mediterranean) in order to assess pollution levels. The isotopic composition was correlated with lead, cadmium, selenium and nickel obtained from the digestive gland and foot of the mussels. Significant negative correlations were found between cadmium, selenium and zinc and the mussel foot, mainly for ¹³C. Significant correlations were also found between lead and cadmium and the digestive gland. Pearson correlations indicated that the ¹³C isotopic signal in foot is a good proxy for the concentration of metals such as lead, cadmium, selenium and zinc. Similarly, ¹⁵N isotopic signatures in the digestive gland reflected the lead and cadmium concentration.     

Field validation,in Scotland and Iceland,of the artificial mussel for monitoring trace metals in temperate seas

The artificial mussel (AM), a novel chemical sampling device, has been developed for monitoring dissolved trace metals in marine environments. The AM consists of Chelex-100 suspended in artificial seawater within Perspex tubing and enclosed with semi-permeable polyacrylamide gel at both ends. To validate the field performance of the AM in temperate waters, we deployed AMs alongside transplanted blue mussels Mytilus edulis in coastal environments in Scotland (Holy Loch, Loch Fyne, Loch Striven and Millport) and Iceland (Reykjavikurhöfn, Gufunes, South of þerney, Hofsvik, Hvalfjörður and Sandgerði) for monitoring trace metals. While uptake patterns of Cd between the AM and M. edulis were highly comparable, discrepancies were found in the accumulation profiles of the other metals (Cu, Cr, Pb and Zn), in particular Zn. Nonetheless, the AMs gave a better resolution to accurately reveal the spatial difference in dissolved metal contamination when compared with M. edulis. AMs complement the use of mussels since AMs indicate dissolved metals in seawater, whereas uptake by mussels indicates a mixture of dissolved and particulate metals. Our results also indicated that historical metal exposure of the transplanted M. edulis could significantly confound their metal concentrations especially when the deployment period was short (i.e. <34 d). This study suggested that the AM can overcome problems associated with variable biological attributes and pre-exposure history in the mussel, and provides a standardized and representative time-integrated estimate of dissolved metal concentrations in different marine environments.     

Effects of trace metal and exposure to air on serotonin and dopamine levels in tissues of the mussel Perna perna

We evaluated levels of serotonin (5HT) and dopamine (DOPA) in muscle and digestive glands of the mussel, Perna perna, collected at different times of day; exposed to air for 24 h, followed by re-submersion; and after exposure to different metals. Mussels collected at different periods of day showed little oscillation in 5HT and DOPA levels. Mussels exposed to metals showed significant changes in 5HT and DOPA levels in digestive gland and muscle, as did mussels exposed to air. Our data suggest that analyses of 5HT and DOPA in tissues of mussels could serve as a tool to evaluate the presence and effects of heavy metal contamination in mussels. Care in data interpretation is required, however, since other environmental factors such as exposure of mussels to air (i.e. at low tides) can also cause changes in DOPA and 5HT levels. Additional research is necessary to separate such natural environmental effects from effects of contaminants.     

Application of artificial mussels (AMs) under South African marine conditions: a validation study

Over the last three decades there has been a significant decline in the number of marine pollution monitoring-related studies in South Africa. Thus, the current study was conducted to assess the current state of metal contamination within the South African marine environment through the validation of the artificial mussel (AM). Indigenous reference mussels (Perna perna) were deployed alongside the passive device within the South African marine environment for a 6 week exposure period. Analysis of metal uptake (Cd, Cr, Cu, Pb and Zn) was determined by filtration and elution of the AM chelex resins, microwave digestion of the transplanted mussels, and determination of their metal concentrations by ICP-MS and ICP-OES analysis. Uptake patterns between the AM and transplanted mussels showed significant temporal and spatial correlation for the majority of the elements analysed. While the AM provided relevant and complementary information on the dissolved metal concentrations, limitations were also observed.     

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.     

Mercury in tissues of mussel off southern California

Total mercury content has been determined in three tissues of the intertidal mussel Mytilus californianus collected at locations along the southern California coast and at island control stations. The apparent influence of certain harbours and municipal wastewater outfalls was reflected by the relatively higher levels of mercury in all tissues of mussels from these areas. In general, the concentrations reported here are lower than those in mussels on west European coasts.     

Comparative petroleum hydrocarbons levels and biochemical responses in mussels from hydrothermal vents (Bathymodiolus azoricus) and coastal environments (Mytilus galloprovincialis)

Aliphatic hydrocarbons and PAHs in the whole soft tissues of Bathymodiolus azoricus from three Mid-Atlantic Ridge hydrothermal vents (Menez-Gwen, Lucky Strike and Rainbow), and Mytilus galloprovincialis from three contaminated coastal sites in South Portugal were analysed, and its effects on the digestive gland microsomes mixed-function oxygenase system (MFO) were assessed.Aliphatic hydrocarbons levels were present in the same magnitude in both coastal and hydrothermal environments, while the UCM (unresolved complex mixture) for coastal mussels were higher than in vent mussels. In general, significantly higher PAHs concentrations were found in coastal mussels, compared to B. azoricus where low molecular weight PAHs (2–3 rings) represented the majority of PAHs contrarily to what was observed in M. galloprovincialis. The MFO components were present in both mussel species, and were detected in vent mussels for the first time. However this system seems to have different roles in species from these contrasting environments. In coastal mussels MFO responded to hydrocarbon contamination while response in hydrothermal organisms appeared to be related mainly to endogenous factors.     

Comparative petroleum hydrocarbons levels and biochemical responses in mussels from hydrothermal vents (Bathymodiolus azoricus) and coastal environments (Mytilus galloprovincialis)

Aliphatic hydrocarbons and PAHs in the whole soft tissues of Bathymodiolus azoricus from three Mid-Atlantic Ridge hydrothermal vents (Menez-Gwen, Lucky Strike and Rainbow), and Mytilus galloprovincialis from three contaminated coastal sites in South Portugal were analysed, and its effects on the digestive gland microsomes mixed-function oxygenase system (MFO) were assessed.Aliphatic hydrocarbons levels were present in the same magnitude in both coastal and hydrothermal environments, while the UCM (unresolved complex mixture) for coastal mussels were higher than in vent mussels. In general, significantly higher PAHs concentrations were found in coastal mussels, compared to B. azoricus where low molecular weight PAHs (2–3 rings) represented the majority of PAHs contrarily to what was observed in M. galloprovincialis. The MFO components were present in both mussel species, and were detected in vent mussels for the first time. However this system seems to have different roles in species from these contrasting environments. In coastal mussels MFO responded to hydrocarbon contamination while response in hydrothermal organisms appeared to be related mainly to endogenous factors.     

Okadaic acid, a causative toxin of diarrhetic shellfish poisoning, in green-lipped mussels Perna viridis from Hong Kong fish culture zones: method development and monitoring

Green-lipped mussels (Perna viridis) were collected from seven fish culture zones (FCZs) in Hong Kong and analyzed for okadaic acid (OA). A conventional HPLC method was modified by incorporating a proteinase K digestion step. Results suggest that a higher recovery (2.5 times higher) of OA was obtained from spiked samples after the addition of 1.08 mg proteinase K in comparison with samples incubated without the proteolytic enzyme. For the hepatopancreas (HP) of individual field-collected mussels, the additional digestion step can enhance OA extraction by 3.1 times. Spatial and temporal variations in OA concentrations in the mussels from various FCZs were investigated. The highest concentration of OA in mussel HP samples was 1164.9 ng/g HP wet wt. With respect to OA concentrations in whole mussel tissues from seven sites and four seasonal samplings, the concentrations were between 70.0 and 131.0 ng/g wet wt., which did not exceed the generally recognized international regulatory criteria (>200 ng/g) for OA.     

Okadaic acid, a causative toxin of diarrhetic shellfish poisoning, in green-lipped mussels Perna viridis from Hong Kong fish culture zones: Method development and monitoring

Green-lipped mussels (Perna viridis) were collected from seven fish culture zones (FCZs) in Hong Kong and analyzed for okadaic acid (OA). A conventional HPLC method was modified by incorporating a proteinase K digestion step. Results suggest that a higher recovery (2.5 times higher) of OA was obtained from spiked samples after the addition of 1.08 mg proteinase K in comparison with samples incubated without the proteolytic enzyme. For the hepatopancreas (HP) of individual field-collected mussels, the additional digestion step can enhance OA extraction by 3.1 times. Spatial and temporal variations in OA concentrations in the mussels from various FCZs were investigated. The highest concentration of OA in mussel HP samples was 1164.9 ng/g HP wet wt. With respect to OA concentrations in whole mussel tissues from seven sites and four seasonal samplings, the concentrations were between 70.0 and 131.0 ng/g wet wt., which did not exceed the generally recognized international regulatory criteria (>200 ng/g) for OA.     

Lysosomal membrane stability and metallothioneins in digestive gland of Mussels (Mytilus galloprovincialis Lam.) as biomarkers in a field study

The lysosomal membrane destabilization and the metallothionein content in the digestive gland cells of mussels (Mytilus galloprovincialis Lam.), collected along the east coast of the North Adriatic (Istrian and Kvarnerine coast, Croatia), were examined over a period of four years (1996–1999). The lysosomal membrane stability, as a biomarker of general stress, showed that the membrane labilization period in mussels from polluted, urban- and industrial-related areas was significantly decreased (p<0.05) when compared to mussels from control, clean sea water sites. In the harbour of Rijeka, the most contaminated site, the lysosomal membrane stability was reduced by more than 70% compared to the control. This method also proved to be a useful biomarker for detection of additional stress caused by short-term hypoxia that occurred once during this study inside the polluted and periodically quite eutrophic Pula Harbour. The concentration of metallothioneins in the mussel digestive gland, as a specific biomarker of exposure to heavy metals, did not reveal significant differences (p<0.05) between sites covered by this study.     

Bioaccumulation and biomarker responses of trace metals and micro-organic pollutants in mussels and fish from the Lagoon of Venice, Italy

The aims of this study were to evaluate levels and effects of trace metals (Cd, Cr, Cu, Fe, Mn, Pb, Zn), PAHs and PCBs in mussels and fish native to the Lagoon of Venice. To this end, bioaccumulation and various biomarkers of generic stress and exposure (malondialdehyde, MDA and metallothioneins, MT; ethoxyresorufin O-deethylase activity, EROD; fluorescent bile metabolites and somatic indices in fish; lysosomal membrane stability, LMS; lipofuscins, LF; neutral lipids, NL and survival-in-air, SOS in mussels) were measured in organisms seasonally collected from two differently influenced areas. In mussels, metal bioaccumulation levels at the two sites were low and fluctuated similarly, exhibiting decreasing levels in summer, like MDA and MT. Micro-organic pollutants and the probably related biological response of LMS did not show either significant site differences or seasonal trends. LF and NL revealed inconclusive patterns. The results of the survival-in-air test were quite erratic, showing that they were associated with the high variability of both natural and physiological parameters temperature, food, reproduction, and body reserve cycle. In fish, both inorganic and organic micropollutant levels were low, and differences were detected between sites only for PCBs. The pattern exhibited by micro-organic contaminants was clearly related to the reproductive cycle at both sites; a relationship with PCBs, EROD and MDA was established at the site where their levels were generally higher. Bile metabolites (3-OH benzo(a)pyrene, 1-OH pyrene) were associated with PAH contents.