Oxygen radicals production and actin filament disruption in bivalve haemocytes treated with benzo(a)pyrene

Haemocytes play an essential role in the internal defence of molluscs. It has been reported that organic xenobiotics commonly found as pollutants in the marine environment impair defence capabilities of haemocytes. The purpose of the present study was to investigate the effects of benzo(a)pyrene [B(a)P] on the integrity of the actin cytoskeleton and on endocytosis in haemocytes and to see if these effects are related to generation of reactive oxygen species. Haemocytes were exposed in vitro to B(a)P (0.5–40 μg/ml) for 1 h. Cell viability (using 2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide or XTT assay) indicated that selected doses were sublethal. Uptake of neutral red was significantly decreased in a dose-dependent manner in B(a)P-treated haemocytes. Distribution of actin filaments, labeled with rhodamine-conjugated phalloidin, was altered in haemocytes treated with 20 or 40 μg/ml B(a)P. These effects could be related to an increased production of superoxide anion during B(a)P metabolism, as detected by the nitroblue tetrazolium (NBT) reduction assay in haemocytes treated with 10 μg/ml B(a)P.     

Cell responses to xenobiotics: comparison of MCF7 multi-drug- and mussel blood cell multi-xenobiotic-defense mechanisms

Multi-drug resistance (MDR) in MCF7 breast cancer cells and multi-xenobiotic resistance (MXR) in mussel (Mytilus edulis) blood cells (MBC) are well known mechanisms that contribute to the decrease in intracellular concentrations of many unrelated but cytotoxic compounds. In the present work, we have carried out comparative investigations of the MDR/MXR protective mechanisms using a rapid colorimetric assay for cell viability and calcein accumulation for MDR/MXR activities. These studies were performed using cultured MCF7 and MBC before and after in vitro exposure to xenobiotics. Our results indicate that a 5-day exposure to doxorubicin or vincristine decreased calcein accumulation in MBC which is consistent with an induction of multi-xenobiotic resistance. The increase in calcein accumulation provoked by 1-h treatment with 50 microM verapamil was much lower in MBC when compared to the P-glycoprotein overexpressing MCF7 cell line. We conclude that such microplate assays could be used in primary cultures of MBC to estimate the effects of various chemicals on MXR activity.     

Effect of biodeposits from mussel culture on macroinvertebrate assemblages at sites of different hydrodynamic regime

The present study examined for differences in macroinvertebrate assemblage composition inside and outside of mussel farm sites experiencing different hydrodynamic regimes. Multivariate analysis revealed that there were significant differences in macroinvertebrate assemblage composition (averaged across seasons) between samples taken inside and outside of the two relatively low energy sites, whilst no such difference was observed for the relatively high energy site. Taxa that best discriminated between the dissimilarities observed in macroinvertebrate assemblage composition inside and outside farms were principally polychaetes (more abundant inside) and ophiuroids (more abundant outside). Sediment total organic matter and number of mussel shells were the parameters that best explained the difference in macroinvertebrate assemblage observed at the two relatively sheltered farm study sites. No taxa or environmental variables were particularly good at discriminating or explaining differences observed inside and outside the relatively high energy farm site. The present study indicates there is a relationship between the hydrodynamic regime of a farm site, organic enrichment of seabed sediments by mussel biodeposits, and a subsequent modification of the macroinverebrate assemblages. This finding has implications for the site selection of mussel farms in coastal environments.     

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.     

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.     

Susceptibility to oxidative stress of mussels (Mytilus galloprovincialis) in the Venice Lagoon (Italy)

The aim of this study was to evaluate the susceptibility to pollutant mediated oxidative stress of the Mediterranean mussel Mytilus galloprovincialis in the Venice lagoon (Italy).

In June 2003, mussels from a farm were transplanted to eight sites in the lagoon for five weeks. Oxidative stress responses were measured by: (i) total oxyradical scavenging capacity (TOSC) assay, for an overall evaluation of the oxidative stress response capability; (ii) catalase (CAT), as a key enzyme involved in the antioxidant defence system; (iii) malondialdehyde (MDA), as an indicator of lipid peroxidation, to evaluate an oxidative damage; (iv) metallothioneins (MTs), as they play a role in the antioxidant defence.

The TOSC analysis revealed a reduced capability to eliminate: (i) peroxyl radical in mussels transplanted at Palude della Rosa, Valle Millecampi and Chioggia; (ii) hydroxyl radical at Campalto and Valle Millecampi; (iii) peroxynitrite at Valle Millecampi.

Inhibition in CAT activity, observed in all the monitored sites, confirms the presence of an oxidative pressure in transplanted mussels.

In addition, Pearson correlation analysis was performed in order to observe possible links between the various parameters. The PCA was a powerful tool to discriminate impacted sites, suggesting that the mussels transplanted throughout the Venice lagoon were subjected to different levels of oxidative pressure. Furthermore, it provided an easy and useful tool to summarize the obtained results.     

Assessment of sacrificial anode impact by aluminum accumulation in mussel Mytilus edulis: a large-scale laboratory test

Since the early 1960s, the application of aluminum alloy sacrificial anodes to mitigate marine corrosion has been well known. The aim of this work was to study aluminum bioconcentration in Mytilus edulis by an in vitro test performed in two tanks: the first containing non-contaminated water (NCW) and the second containing aluminum-contaminated water (CW) (530 μg L⁻¹) released by sacrificial anode. The mussels were collected and examined over a period of 8 weeks. A comparison between the aluminum concentrations in the digestive glands of mussels from the CW and NCW tanks shows that the highest value (1700 mg/kg d.w.) was found in the CW mussels collected after 13 days. In NCW, the mean aluminum concentration in digestive glands during the test was 281 mg/kg d.w. The rapid concentration decrease in digestive glands is probably due to the inhibition of filtering activity due to valve closure at the high concentration as well as the induction of the detoxification response.     

Influence of climate change on interannual variation in contaminant body burden in Gulf of Mexico oysters

As part of NOAA's National Status and Trends Mussel Watch Program, oysters were sampled along the Gulf of Mexico coast each winter from 1986 to 1992 and analyzed for trace metal, polynuclear aromatic hydrocarbon (PAH), and pesticide body burden. We identified contaminant variables for which large-scale climate processes played an important role in establishing the interannual variation in body burden by examining cases where body burdens rose or fell more or less in unison over broad geographic regions and distinguished these concordant changes from cases where bays varied independently. Of the 11 metals analyzed, nine had scales of concordancy 100 km. Ni and Se, however, had among the largest scales of concordancy in the study, 1200 km. That is, oyster body burdens in bays as far apart as 1200 km tended to rise and fall in unison from one year to the next. Interannual variations in body burden of organic contaminants had a much stronger regional component. All but two of the 11 organic contaminants had scales of concordancy of 200 km or greater and six exceeded 400 km. Concordancy was strongest either in the southern, northwestern, or north-central Gulf, depending upon the contaminant. For all contaminants, bays tended to vary independently in the northeastern Gulf. For three contaminants, total chlordanes, dieldrin and Cd, regional concordancy may originate from a widespread decrease in use and, therefore, input. These contaminants declined nearly monotonically over the 7 years. For others, including Zn and many of the PAHs and pesticides, the El Niño/Southern Oscillation (ENSO) cycle may be important in establishing the interannual variability in contaminant body burden: (1) ENSO has the geographic scale required; (2) a subtropical influence is required to explain the similarity in interannual variation between south Texas and southern Florida; (3) ENSO-related climate responses in the Gulf follow the southwestern/northeastern trend that would establish the northwestern Gulf focus in concordancy so prominent in many of the contaminants; and (4) deviations in yearly mean body burden from the Gulf-wide mean track the Southern Oscillation Index in some cases.     

紫贻贝(Mytilus edulis)对4种重金属的生物富集动力学特性研究

应用半静态双箱模型室内模拟了紫贻贝对 4种重金属 (As、Hg、Cd、Pb)的生物富集实验 ,通过非线性拟合得到了紫贻贝富集重金属的吸收速率常数k1 、排出速率常数k2 、生物富集因子BCF、生物学半衰期B1 / 2 等动力学参数。实验海水盐度为 32、温度分别为( 1 2 .34± 2 .2 8)℃和 ( 6.63± 3.1 2 )℃、溶氧量 >6.5mg/L。得到的Hg、Cd、Pb的动力学参数k1为 8.71 2— 30 1 .5 ;k2 为 0 .0 0 942 5— 0 .0 30 64;BCF为 783.4— 1 1 75 4 ;B1 / 2 为 2 2 .62— 73.5 4。比较结果得出 :吸收速率常数k1 及生物富集因子BCF均随着外部水体浓度的增加而减小 ,在平衡状态下生物体内金属含量CAmax随着外部水体浓度的增加而增加 ,且基本呈正相关 ,说明紫贻贝是比较理想的重金属Hg、Cd、Pb污染的指示生物。紫贻贝排出Cd的生物学半衰期要长于金属Hg、Pb、As。     

Market driven management of climate change impacts in the Spanish mussel sector

Mussel farming is the largest aquaculture activity in Spain both in volume and value, being a key pillar for the development of the coastal areas in Galicia (NW Spain), where this production is concentrated. The available scientific knowledge suggests that in the long term the primary productivity of the Galician estuaries will be reduced due to climate change. Consequently, adaptive management will be required to face this challenge. The impact of the likely production decrease will depend, among other factors, on the availability of substitutes, being their identification the main object of this research. So, it was analysed whether this markets are integrated or not through cointegration tests, finding that only the French and the Spanish markets are partially integrated. This implies that since very little substitutes are available for the Galician fresh mussels, decreases in production will not necessarily affect producer's income, as prices increases will compensate the (eventual) reduction in volume. In terms of policy, this study suggests that market intelligence may contribute to an adaptive, pragmatic and flexible framework to face climate change impacts, avoiding too anticipated or blind responses that may result in economic, environmental and social costs.