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.     

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.     

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.     

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.     

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.     

Evaluation of the Mytilus galloprovincialis Lam. digestive gland metallothionein as a biomarker in a long-term field study: seasonal and spatial variability

A study was made of seasonal and spatial variability of metallothionein (MT) concentrations, determined spectrophotometrically in the soluble fraction of the digestive gland of mussels Mytilus galloprovincialis, collected between 1999 and 2001 from several coastal and estuarine locations along the central Eastern Adriatic coast (Croatia). The seasonal influence on the MT and metal concentrations (higher values in winter-spring season, than in summer-autumn season) is more pronounced than the local site-specific influence. Furthermore, within each season a significant site-specific dependence on the MT and trace metal variations can be detected. An inverse relationship of mussel condition index (CI) and temperature with MT and trace metals levels indicates the influence of food abundance and mussel annual reproductive cycle. Substantially higher concentrations of both MT and Cd were recorded in mussels inhabiting estuarine locations, possibly indicating a markedly higher Cd bioavailability at these locations. The positive correlations obtained between MT and Cd in all seasons except autumn support an argument for application of digestive gland MT as a biomarker of Cd exposure, providing evidence for assessing the most appropriate season for mussel sampling.     

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.     

Bioavailability of heavy metals in soil of the Tieguanyin tea garden,southeastern China

The bioavailability of 22 heavy metals was investigated at 19 sampling sites in Tieguanyin tea garden in Anxi County,Fujian Province,southeastern China.Heavy metal concentrations were determined by inductively coupled plasma-mass spectrometry(ICP-MS)and evaluated by geo-accumulation index(I_(geo)).Dilute nitric acid extraction was used to evaluate biological activity.Cu,Pb,and Cd were highly bioavailable and most easily absorbed by tea trees.Heavy metal bioavailability in the surface soil was as the ratio of the effective state to the total amount.Cd had the highest I_(geo)values,and the respective samples and sites were classified as moderately/strongly contaminated.Cd element is considered the main factor of heavy metal pollution in the tea garden in Anxi.The other heavy metals studied were present in lower concentrations;thus,the samples were classified as uncontaminated or slightly contaminated.     

Histopathology alterations and histochemistry measurements in mussel, Mytilus edulis collected offshore from an aluminium smelter industry (Norway)

Histopathological characteristics of specific organs express condition, and represent time-integrated impacts on the organism stemming from alterations at lower levels of biological organisation. As integrative parameters, histochemical investigations have proved to be sensitive tools to detect effects of chemical compounds.The objective of this study was to determine changes in the tissues of mussels collected at a PAH contaminated site compared to a reference site using histopathological and histochemical parameters: lipofuscin (LF) accumulation in mussel digestive gland, and lysosomal membrane stability (LMS), and using additional information provided by body burden analysis to compare the sensitivity of these parameters.The histochemical measurements for both LF and LMS gave a clear indication of a high level of stress in animals from the PAH contaminated site. This LF accumulation in lysosomes is the result of peroxidation of autophagocytosed proteins associated with protein aggregates and oxidatively damaged organelles. These measurements were able to detect the effects of PAHs, and showed a strong relationship with the body burden results.     

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.     

Histopathology alterations and histochemistry measurements in mussel,Mytilus edulis collected offshore from an aluminium smelter industry (Norway)

Histopathological characteristics of specific organs express condition, and represent time-integrated impacts on the organism stemming from alterations at lower levels of biological organisation. As integrative parameters, histochemical investigations have proved to be sensitive tools to detect effects of chemical compounds.The objective of this study was to determine changes in the tissues of mussels collected at a PAH contaminated site compared to a reference site using histopathological and histochemical parameters: lipofuscin (LF) accumulation in mussel digestive gland, and lysosomal membrane stability (LMS), and using additional information provided by body burden analysis to compare the sensitivity of these parameters.The histochemical measurements for both LF and LMS gave a clear indication of a high level of stress in animals from the PAH contaminated site. This LF accumulation in lysosomes is the result of peroxidation of autophagocytosed proteins associated with protein aggregates and oxidatively damaged organelles. These measurements were able to detect the effects of PAHs, and showed a strong relationship with the body burden results.     

Histopathology alterations and histochemistry measurements in mussel, Mytilus edulis collected offshore from an aluminium smelter industry (Norway)

Histopathological characteristics of specific organs express condition, and represent time-integrated impacts on the organism stemming from alterations at lower levels of biological organisation. As integrative parameters, histochemical investigations have proved to be sensitive tools to detect effects of chemical compounds. The objective of this study was to determine changes in the tissues of mussels collected at a PAH contaminated site compared to a reference site using histopathological and histochemical parameters: lipofuscin (LF) accumulation in mussel digestive gland, and lysosomal membrane stability (LMS), and using additional information provided by body burden analysis to compare the sensitivity of these parameters. The histochemical measurements for both LF and LMS gave a clear indication of a high level of stress in animals from the PAH contaminated site. This LF accumulation in lysosomes is the result of peroxidation of autophagocytosed proteins associated with protein aggregates and oxidatively damaged organelles. These measurements were able to detect the effects of PAHs, and showed a strong relationship with the body burden results.     

Accumulation of Metals in Naturally Grown Weeds (Aquatic Macrophytes) Grown on an Industrial Effluent Channel

The ability of seven hyperaccumulator macrophytes which grow naturally in the heavy metal contaminated channels of three different industries (Hindustan Aeronautical Ltd., Eveready Ltd., and Scooter India Ltd.) to accumulate heavy metals was recorded. All these industries use electroplating processes in their manufacturing and are located in the inner area of Lucknow City, U.P., India. Of the three industries monitored, effluent released from Eveready Ltd. contained the highest concentration of heavy metals. In general, accumulations of heavy metals depend upon the plant species and the metal concentration in the media. All plant samples showed heterogeneous metal accumulations, except for Fe or Cd. It was observed that some plant species accumulated high level of metals, e. g., Eichhornnia crassipes for Fe (4052.44 μg/g), Mn (788.42 μg/g), and Cu (315.50 μg/g), and Spirodela polyrhiza for Cd (12.75 μg/g), Pb (20.25 μg/g), and Cr (128.27 μg/g), even when the metal concentrations were not high in the effluent. In summary, these two plants were found to be the best accumulators at each contaminated site. The results will be helpful in the selection of plant species which can be used as bioaccumulators or bioindicators.     

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.     

Sediment chemical contamination of a shallow water area close to the industrial zone of Porto Marghera (Venice Lagoon,Italy)

The industrial zone of Porto Marghera is one the most important “contaminated sites of national interest” (SIN) in Italy, being identified as an area of high environmental risk. The site includes a wide shallow water area of the Venice Lagoon extending toward the city of Venice, which was investigated in order to acquire information on the pollutant level and distribution. Grain-size, heavy metal, nutrient, and organic micropollutant concentrations were determined in the surface sediment layers (0–5 and 5–10 cm depths) of 51 sites. A generally low contamination was found, except for Hg concentration, which increases the toxicological risk in most of the sites of the area, according to the results of a comparison with Sediment Quality Guidelines. A heavy pollution fingerprint (Cd, Cu, Hg, Pb, and Zn up to 15.2, 257, 11.9, 248, and 3010 mg/kg d.w., respectively) was instead found near the Tresse Island, which is ascribed to the spill of pollutants from the contaminated sediment disposed therein. Grain-size and heavy metal profiles down to a depth of 40 cm in eight selected sites, finally show a probable decrease of the pollution affecting the area in recent years.     

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.     

Sediment chemical contamination of a shallow water area close to the industrial zone of Porto Marghera (Venice Lagoon, Italy)

The industrial zone of Porto Marghera is one the most important "contaminated sites of national interest" (SIN) in Italy, being identified as an area of high environmental risk. The site includes a wide shallow water area of the Venice Lagoon extending toward the city of Venice, which was investigated in order to acquire information on the pollutant level and distribution. Grain-size, heavy metal, nutrient, and organic micropollutant concentrations were determined in the surface sediment layers (0-5 and 5-10 cm depths) of 51 sites. A generally low contamination was found, except for Hg concentration, which increases the toxicological risk in most of the sites of the area, according to the results of a comparison with Sediment Quality Guidelines. A heavy pollution fingerprint (Cd, Cu, Hg, Pb, and Zn up to 15.2, 257, 11.9, 248, and 3010 mg/kg d.w., respectively) was instead found near the Tresse Island, which is ascribed to the spill of pollutants from the contaminated sediment disposed therein. Grain-size and heavy metal profiles down to a depth of 40 cm in eight selected sites, finally show a probable decrease of the pollution affecting the area in recent years.     

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.     

Phytoremediation of heavy metal‐contaminated water and sediment by Eleocharis acicularis

Phytoremediation is an environmental remediation technique that takes advantage of plant physiology and metabolism. The unique property of heavy metal hyperaccumulation by the macrophyte Eleocharis acicularis is of great significance in the phytoremediation of water and sediments contaminated by heavy metals at mine sites. In this study, a field cultivation experiment was performed to examine the applicability of E. acicularis to the remediation of water contaminated by heavy metals. The highest concentrations of heavy metals in the shoots of E. acicularis were 20 200 mg Cu/kg, 14 200 mg Zn/kg, 1740 mg As/kg, 894 mg Pb/kg, and 239 mg Cd/kg. The concentrations of Cu, Zn, As, Cd, and Pb in the shoots correlate with their concentrations in the soil in a log‐linear fashion. The bioconcentration factor for these elements decreases log‐linearly with increasing concentration in the soil. The results indicate the ability of E. acicularis to hyperaccumulate Cu, Zn, As, and Cd under natural conditions, making it a good candidate species for the phytoremediation of water contaminated by heavy metals.     

Heavy metals in Baltic herring and cod

The monitoring programme of harmful substances in selected species in the Baltic Sea was started in 1979. In the present study, three-year averages of Zn, Cu, Cd and Pb in cod liver (Hg in cod muscle) and herring muscle are reported. The concentrations of heavy metals in cod show areal differences roughly in accordance with the concentrations of heavy metals in sea water. No significant differences between heavy metal concentrations in North Atlantic and Baltic Sea species were found, except for mercury, for which values in the Baltic Sea species are lower than in those from the North Atlantic.