Integrated use of biomarkers (superoxide dismutase, catalase and lipid peroxidation) in mussels Mytilus galloprovincialis for assessing heavy metals' pollution in coastal areas from the Saronikos Gulf of Greece

Mussels are used as sentinel organisms and bioindicators to evaluate the toxic effects of chemical pollutants in marine organisms, especially heavy metals, representing an important tool for biomonitoring environmental pollution in coastal areas. Antioxidant defence enzymes play an important role in cellular antioxidant defence systems and protect from oxidative damage by reactive oxygen species (ROS). Indigenous mussels Mytilus galloprovincialis of the Saronikos Gulf of Greece were used for monitoring heavy metal pollution in three polluted sites in the area and in one unpolluted site. Seasonal variations of the activity of antioxidant defence enzymes, superoxide dismutase (SOD) and catalase (CAT), as well as lipid peroxidation (LP) were measured as biomarkers in a period of three years in relation to concentrations of trace metals in their gills and mantle and compared to mussels from an unpolluted sampling site. SOD activity increased at least 2 fold at the polluted sites when compared to the control site (the high activity was recorded in the spring time). CAT activity was increased 2-3 times at the polluted sites, with high activity in the winter and spring time, compared to the control site. LP concentration was twice higher at the polluted sites, following the same seasonal pattern. Trace metals contents in mussels collected at polluted sites were 3-4 fold higher compared to the control site and showed moderate variations along the months, with a winter maximum followed by a summer pre-spawning minimum matching the seasonal trends of temperature and salinity. Our results showed that metal pollution in the Elefsis Bay (the most polluted coastal area) causes relatively medium levels of oxidative stress in tissues of mussels due to cellular oxy-radical generation. This study, which is the first in the area, showed that seasonal variations of the activity of antioxidant defence enzymes and LP concentrations in mussels can be used as potential biomarkers of toxicity for long-term monitoring in marine coastal ecosystems.     

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.     

The influence of diet on comparative trace metal cadmium,copper and zinc accumulation in Thais clavigera (Gastropoda: Muricidae) preying on intertidal barnacles or mussels

The influence of diet on comparative metal accumulation was investigated using a predatory muricid gastropod Thais clavigera. Individuals were fed for up to 56 days on either barnacles, i.e., Tetraclita squamosa, or mussels, i.e., Perna viridis, collected from metal-contaminated and clean sites. Barnacles and mussels have contrasting metal handling strategies and, therefore, different body concentrations, intracellular distributions and detoxification systems. Field collection of prey items that accumulated body metal concentrations over a lifetime of exposure allowed bioavailability to the predator, T. clavigera, to be assessed naturally, which may not be the case for prey exposed to metals for a short time in the laboratory. T. clavigera that was fed cadmium- and copper-contaminated barnacles or mussels ingested significantly greater amounts compared to those fed conspecifics collected from clean locations. T. clavigera body cadmium and copper concentrations were not, however, significantly different between individuals fed either contaminated or clean prey. Amount of zinc ingested was similar in mussels collected from clean and contaminated environments but much less when compared to the barnacle prey. The body concentrations of zinc in T. clavigera fed mussels collected from both sites fell. In contrast, the amount of zinc ingested from barnacle prey was significantly greater from those collected from the metal-contaminated site as compared to the clean one. This was reflected as significantly greater body zinc concentrations in T. clavigera fed contaminated barnacles compared to those fed clean individuals. Copper and zinc accumulation from prey was, therefore, complex. It varied between metal and between prey type, but appeared to be related to the amount ingested and the metal handling strategy of the prey.     

Selection of bioindicators for monitoring marine environmental quality in the Bay of Fundy,Atlantic Canada

Distribution of metals, PAH's and PCB's in lobsters, mussels, and sediments were used to assess marine environmental quality of the Bay of Fundy. This study demonstrates that the lobster (Homarus americanus) is a better bioindicator for monitoring contaminants in the marine environment and has a greater capacity for the uptake and accumulation of contaminants than the mussel (Mytilus edulis) and sediments. A definite pattern in the spatial distribution of lobster Cu, Cd, and Ag was evident. The distribution of organic contaminants for both mussels and lobsters in the Bay of Fundy lacked a spatial trend, and organic contaminants were undetectable in sediments from all sites. The Gulf Watch Programme, which monitors chemicals in mussels in the Bay of Fundy, did not indicate a problem with high levels of Cu, Cd, and Zn in the ecosystem. Analytes below the detection limit, such as in mussels and sediments, increase the difficulties of chemical analysis and detection for environmental monitoring. Deficiencies of mussels in monitoring the Bay of Fundy were discussed.     

Cornish mackerel

Port Phillip Bay receives the industrial and domestic effluents from Melbourne and a recent survey revealed an accumulation of a variety of heavy metals in bottom sediments. These studies have now been extended to survey the accumulation in oysters and mussels, and in most areas in the Bay these bivalves are heavily contaminated with cadmium. Oysters accumulate more of this metal than mussels.     

基于"标准化"背角无齿蚌(Anodonta woodiana)对太湖五里湖重金属污染的主动监测

将人工繁育的"标准化"背角无齿蚌(Anodonta woodiana)移殖至太湖五里湖,并以仍养殖在未受污染的中国水产科学研究院淡水渔业研究中心南泉基地的同批蚌作为对照,进行为期9个月的主动监测研究.每3个月回收一次蚌样,应用电感耦合等离子质谱仪(ICP-MS)测定15种重金属(Al、Cr、Mn、Fe、Co、Ni、Cu、Zn、As、Mo、Ag、Cd、Ba、Tl和Pb)的含量.结果表明,南泉基地对照组和五里湖移殖组的蚌样对重金属均产生了明显的生物积累.培养3个月的五里湖移殖组蚌样中As的含量显著高于同期南泉基地对照组,而前者Mn、Fe、Zn和Ba含量显著低于后者;培养6个月的五里湖移殖组蚌样中Al和Pb的含量显著低于同期南泉基地对照组;培养9个月的五里湖移殖组蚌样中Pb含量显著低于同期南泉基地对照组.然而,南泉基地对照组和五里湖移殖组蚌样中重金属(Cr、Cu、As、Cd和Pb)含量均低于我国及国际上的相关标准.培养3、6和9个月的南泉基地对照组及五里湖移殖组蚌样的重金属污染指数分别为1.8、1.8,2.4、2.1和8.3、16.8,均值综合污染指数分别为0.0218、0.0289,0.0337、0.0218和0.0560、0.0732,属于正常背景水平,并且两水体蚌样的重金属污染指数和均值综合污染指数无显著差异,提示五里湖和南泉基地均未受到明显的重金属污染.     

Izmit Bay (Turkey) ecosystem after Marmara earthquake and subsequent refinery fire: the long-term data.

As a part of Marmara Sea, Izmit Bay (Turkey) has been one of the most polluted sites in the region for the last 25 years. On 17 August 1999, a powerful earthquake along the North Anatolian Fault struck the eastern part of the Marmara region including Izmit Bay. The earthquake destroyed many coastal cities. The Bay was also affected by the quake and subsequent fire in the refinery situated on the north-eastern coast of the Bay. Oceanographic characteristics and polycyclic aromatic hydrocarbon (PAH) levels of Izmit Bay (Marmara Sea) have been investigated to find out the degree of contamination. Seawater samples were collected at nine stations of the Bay in April and September 1999 and the results were compared with those obtained in the previous years (1984 and 1994). Monitoring data are presented for plant nutrients (nitrate + nitrate, ortho-phosphate and silicate), dissolved oxygen and chlorophyll a. Surface sediments and mussels, Mytilus galloprovincialis, have been analysed for total PAH (T-PAH) contents in April and September 1999 (before and after the Marmara Earthquake) for the samples collected from eight coastal stations of the Bay. Biomarker (Lysosomal stability and feeding rate) studies at three different sites of the Bay have also been performed to investigate the effect of pollution on mussels. Nitrate + nitrite levels in the upper layer of the eastern part of the Bay increased significantly compared to those measured before the earthquake. Of the samples analysed, the highest o-phosphate concentrations were found in September 1999 in the bottom waters of the Bay. The concentration of chlorophyll a reached its minimum value of the last 15 years. Dissolved oxygen decreased dramatically from 1984 to 1999. Total PAH concentrations measured in April 1999 at both offshore and coastal sites of the Bay were more or less the same (2 micrograms l-1). The subsequent fire after the earthquake caused an increase in the total PAH levels in water column, in sediment and in mussels. Seawater total PAH concentrations ranged between 3.5 and 11 micrograms l-1 at open coast stations and 5-17.5 micrograms l-1 at coastal stations in September 1999. A 2- to 3-fold increase in sediment PAH concentrations (200-5220 mg kg-1 dry weight) was detected after the earthquake. This increase was much more significant in the sediments located around the refinery. More contaminated mussels were detected around the refinery area (110-170 mg kg-1 dry weight). Overall, sediment and mussel PAH concentrations in Izmit Bay are much higher than those found in the other marine systems. In general, the feeding rate and the neutral red retention times of the mussels decreased in some sites of the Bay after the earthquake, but no direct correlation could be detected between the body burden of mussels and biomarkers or between the two biomarker techniques.     

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.     

Petrowatch: Petroleum hydrocarbons,synthetic organic compounds,and heavy metals in mussels from the Monterey Bay area of central California

Mussel Watch techniques were used to measure the concentrations of petroleum hydrocarbons, synthetic organic hydrocarbons, and trace metals in a preliminary study of the Monterey Bay area in central California. Resident mussels were found to have higher-than-expected petroleum hydrocarbon body burdens in Carmel Bay, an area thought to be relatively contaminant free. A ‘hot spot’ of PCB 1254 and pp′DDE was measured at Año Nueuvo Island, a site previously recognized because of elevated levels of mercury in mussels. PCB 1254 concentrations at Año Nuevo Island were high in relation to the remainder of coastal United States and Baja California, reflecting entrainment of this compound at a biologically active area. The Monterey Harbor Jetty location showed the highest contamination levels for zinc and lead. In fact, this location has the second highest lead concentrations in mussels for the California coast. Biological cycling of certain compounds (PCB, DDT and mercury) in remote locations such as Año Nuevo Island, is hypothesized as a mechanism for these elevated concentrations in mussels. Levels of other synthetic organic hydrocarbons were generally low, with chlordanes showing higher concentrations in the more urbanized locations of the study area, i.e. the Monterey Harbor Jetty.     

Alkylbenzenes in mussels from South and South East Asian coasts as a molecular tool to assess sewage impact

Alkylbenzenes, molecular markers of sewage, were measured in 34 green mussels collected from India, Indonesia, Malaysia, Thailand, Cambodia, Vietnam, and the Philippines together with blue mussels collected from Tokyo Bay, Japan. Linear alkylbenzene (LAB) concentrations in South and South East Asian countries ranged from 10 to 1640 ng-∑LAB/g-dry tissue. In some populous cities, LAB concentrations were similar or higher than those found in northern Tokyo Bay which is heavily impacted by sewage effluents. I/E ratios (a ratio of internal to external isomers of LABs) in the South and South East Asian countries (1–3) were much lower than those in Tokyo Bay (3–8), indicating sewage discharged in the coastal zone is poorly treated (e.g., raw sewage and/or primary effluents). Alkylbenzenes with branched alkyl chains, tetrapropylene-based alkylbenzenes, were also detected in mussels from Indonesia and Philippines. This “tell-tale” sign indicates that poorly degradable detergents are still in use in this area, although they have long been phased-out in many industrialized countries.     

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.     

Stable carbon and nitrogen isotope signatures indicate recovery of marine biota from sewage pollution at Moa Point,New Zealand

Stable carbon and nitrogen isotopes have been used to assess sewage contamination of a sewage outfall, discharging milli-screened effluent into Moa Point Bay, New Zealand, and monitor the recovery of flora and fauna after the outfall's closure. An initial study characterising the extent of the discharge and the effects on seaweed (Ulva lactuca L.), blue mussels (Mytilus galloprovincialis) and limpets (Cellana denticulata) from the area, showed effects of the sewage discharge on flora and fauna were localised within in the bay. The immediate area surrounding the discharge area was found to contain limited biodiversity, with an abundance of Ulva lactuca, a bright green lettuce-like seaweed, typically found in areas with high nutrient input, limpets and small blue mussels. The nitrogen isotopic signature (delta15N) is shown to be a good tracer of sewage pollution in seaweed and associated grazers (i.e. limpets) as a result of the increased contribution of urea and ammonia to seawater nitrogen derived from the effluent. The carbon isotopic signature (delta13C) is suggested as a more appropriate sewage tracer for mussels, which filter feed the effluent's particulate organic matter from the water. Lower carbon:nitrogen ratios were found in Ulva lactuca sampled from around the outfall region compared to uncontaminated control sites. However carbon:nitrogen ratios do not vary significantly amongst shellfish species.After closure, monitoring continued for 9 months and showed that the carbon and nitrogen isotopic signatures of algae (Ulva lactuca L.) returned to similar control site levels within 3 months. Limpet and blue mussels (Cellana denticulata and Mytilus galloprovincialis) showed slower recovery times than the Ulva lactuca, with detectable levels of the sewage-derived carbon and nitrogen remaining in the animal's tissue for up to 9 months.     

The usefulness of transplantation studies in monitoring of metals in the marine environment: South African experience

Due to their close proximity to the point sources of pollution, estuaries and harbours are exposed to metals. Mussels are used worldwide to monitor marine pollution due to their ability to take up contaminants and the ease of transplantation. Mussels were collected from two reference sites and transplanted in Richards Bay Harbour (2006 and 2010) and in three harbours. Transplanted and resident mussels were removed after 6 weeks exposure and metal concentrations were analysed using ICP-MS techniques. Transplanted mussels had higher metal concentrations than the resident mussels. This was attributed to regulation of metals by the resident mussels. Metal regulation was greater in mussels that are continuously submerged as opposed to those that undergo tidal influences. For monitoring it is essential that the reference population for transplantation is considered carefully as upwelling events in the pristine reference location results in significant increases in metal bioaccumulation.     

An ecotoxicological approach for the Boka Kotorska Bay (south-eastern Adriatic Sea): first evaluation of lysosomal responses and metallothionein induction in mussels

Mytilus galloprovincialis was used as a biomonitoring organism in Boka Kotorska Bay, a coastal transitional ecosystem in south-western Montenegro. Native mussels were collected in June 2008 at four sites thought to be differently impacted. Biological effects were investigated analysing both generic and specific biomarkers at cellular level (metallothionein content, lysosomal membrane stability, lipofuscin and neutral lipid accumulation, lysosomal structural changes). Trace element levels in mussels were quite low, only Cu and Zn exhibiting slight increases in the two sampling sites of Tivat Bay, the first one in front of a small working dockyard and the other in the water area of a former Naval dockyard. Mussels collected from these sites exhibited the highest values in neutral lipid and lysosomal volume density and the lowest neutral red retention times. Metallothionein content was always low, reflecting the minor body burden of the trace elements and suggesting a non-relevant environmental induction.     

Ecotoxicity assessment of natural attenuation effects at a historical dumping site in the western Baltic Sea

During the late 1950s and early 1960s of the past century, industrial waste material highly enriched in various contaminants (heavy metals, PAHs) was dumped in the inner Mecklenburg Bay, western Baltic Sea. Large-scale shifts in the spatial distribution of heavy metals in surface sediments were mapped by geochemical monitoring in the mid-1980s and 12 years later in 1997. A further study in 2001 was designed to investigate the small-scale spatial distribution of contaminants inside, on top of, and around the historical dumping ground and to examine possible effects to benthic organisms (Arctica islandica, microbiological toxicity tests). The site is located within an area characterized by a discontinuous deposition of fine sediments, but net sedimentation rate of about 3mm/y led ultimately to a gradual coverage of the dumped material. During the first decades after the dumping, about half of the originally dumped material was already spread over the surrounding area as a result of sediment dynamics and re-suspension processes. Recent sediments of the historical dump site are still significantly enriched in heavy metals and PAHs. Microbiological toxicity test results were positive, though uptake of contaminants in mussels was found to deviate only slightly from that of a non-contaminated reference station due to a gradual dilution and decrease in bioavailability of the contaminants.     

An ecotoxicological approach for the Boka Kotorska Bay (south-eastern Adriatic Sea): First evaluation of lysosomal responses and metallothionein induction in mussels

Mytilus galloprovincialis was used as a biomonitoring organism in Boka Kotorska Bay, a coastal transitional ecosystem in south-western Montenegro. Native mussels were collected in June 2008 at four sites thought to be differently impacted. Biological effects were investigated analysing both generic and specific biomarkers at cellular level (metallothionein content, lysosomal membrane stability, lipofuscin and neutral lipid accumulation, lysosomal structural changes). Trace element levels in mussels were quite low, only Cu and Zn exhibiting slight increases in the two sampling sites of Tivat Bay, the first one in front of a small working dockyard and the other in the water area of a former Naval dockyard. Mussels collected from these sites exhibited the highest values in neutral lipid and lysosomal volume density and the lowest neutral red retention times. Metallothionein content was always low, reflecting the minor body burden of the trace elements and suggesting a non-relevant environmental induction.     

Seasonal variations of physiological and cellular biomarkers and their use in the biomonitoring of north Adriatic coastal waters (Croatia)

Lysosomal membrane stability, the content of neutral lipids and lipofuscin as cellular biomarkers were measured in the digestive gland of mussels, Mytilus galloprovincialis Lam., during a one year survey at selected sites along the coast of the northern Adriatic. The ability of mussels to survive in air as a general physiological response was tested at the same sites. Annual changes of sea temperature, salinity, O(2) saturation, pH and chlorophyll a concentration were measured and the effects of these environmental factors on the biomarkers selected for this study were determined. Survival of mussels after aerial exposure was significantly (p < 0.05) reduced in mussels collected from the contaminated harbors of Pula and Rijeka compared to clean control sites. Lysosomal membrane stability of mussels from urban and industrial areas was also lower indicating exposure to different environmental contaminants. At the same sites, the content of neutral lipids and lipofuscin in digestive cells was significantly higher compared to the reference sites, suggesting alterations in lipid metabolism and contaminant-induced peroxidation of cell membranes. The greatest influence of natural environmental factors was observed for the aerial survival of mussels and the neutral lipids and lipofuscin content in the digestive gland, indicating that the annual cycles of biomarker responses should be well understood for a correct interpretation of biomonitoring results.     

Cytological and Physiological Biomarker Responses from Green Mussels, Perna viridis (L.) Transplanted to Contaminated Sites in Hong Kong Coastal Waters

Biological monitoring of marine contaminants often involves the analysis of metals accumulated in mussel tissues. Critiques of this type of monitoring suggest that although relatively good indications of ambient metal concentrations are determined, actual biological harm is not elucidated. Biomarkers, techniques originally developed in medicine, have recently been employed in an attempt to ascertain the health of marine mussels. Biomarkers have been used with good effect in temperate marine pollution monitoring programmes. The use of biomarkers in the toxicological evaluation of tropical and subtropical waters is, however, generally lacking, which is somewhat surprising, as considerable industrialisation and environmental degradation is taking place in the Pacific-rim countries of Southeast Asia. In the present study, a suite of cytological and physiological biomarkers have been measured from mussels (Perna viridis: Mytilidae) transplanted within the coastal waters of Hong Kong. Mussels from contaminated sites, had labile lysosomal membranes, whereas animals from a cleaner reference site had more stable membranes. These cytological responses have been related to metal-induced stress and are simple, cost-effective markers of pollution. Cardiac activity from mussels transplanted to polluted sites was generally similar to that of individuals from the reference site, although a mild tachycardia was associated with one site. The cause of the tachycardia and the use of heart rate in mussels as a biomarker is discussed. Integrated physiological responses to stress, condition indices, revealed mussels from a clean reference site had higher indices than animals from contaminated sites. The relative sensitivities of the cytological and physiological biomarkers deployed in this study and their application to pollution monitoring is discussed.     

Old cannery wastes,a potential source of trace metals in the marine environment

High levels of lead and zinc have been measured in the California Mussel Mytilus californianus taken from southern Monterey Bay as part of the California State Mussel Watch programme. The levels of lead and zinc increase steadily from Cypress Point towards Cannery Row. The California State Water Resources Control Board (which funded but did not conduct the State Mussel Watch) has attributed the high levels to heavy metals contributed by sewage. The state mussel watch data reports have not considered the historic canning wastes from more than 30 canneries along Cannery Row as a possible source of trace metals. Leachates from old cannery wastes may be a major source of lead and zinc in mussels in southern Monterey Bay and must be considered in assessing any data from the State Mussel Watch programme.     

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.