Application of the biomarker responses in scallop (Chlamys farreri) to assess metals and PAHs pollution in Jiaozhou Bay,China

Scallops (Chlamys farreri) were collected at three sites in the Jiaozhou Bay, China in April and July 2007. The responses of biomarkers were evaluated for assessment of physiological status of scallops and pollution in the areas. Compared with site S1, the activity of aryl hydrocarbon hydroxylase was induced notably (P < 0.05) and glutathione S-transferase activity was inhibited significantly (P < 0.05) in S2 and S3. The levels of DNA alkaline unwinding, DNA protein crosslinks, protein carbonyl and lipid peroxidation were tested and indicated the oxidative stress situation of scallops. There was significant difference of biomarker levels between sampling seasons (P < 0.05), and between digestive gland and gill (P < 0.05). The results provided the reference data for multiple-pollution assessment in the marine environment and indicated that tissue type and seasons affect biomarkers, therefore these factors should be taken into consideration when biomarkers are used for environmental assessment purposes.     

A multi-biomarker approach to assess effects of Triclosan in the clam Ruditapes philippinarum

The effects of 7 days’ exposure to differing Triclosan (TCS) concentrations (300, 600, and 900 ng/L) were investigated in the clam Ruditapes philippinarum. Vitellogenin (Vg)-like protein levels in haemolymph and digestive gland from males and females, gill acetylcholinesterase (AChE) activity, superoxide dismutase (SOD) and catalase (CAT) activities in gills and digestive gland, and gill lipid peroxidation (LPO) were measured. The highest TCS concentrations decreased significantly Vg levels in male haemolymph and digestive gland, whereas no significant variations were found in females. The highest TCS concentrations increased significantly SOD activity in gills, but decreased it in digestive gland. No changes in CAT activity were observed. In gills, TCS reduced significantly AChE activity, but it did not induce significant variations in LPO. Our study demonstrates that TCS alters biochemical parameters in R. philippinarum, even at environmentally realistic concentrations, and suggests differing modes of action of the contaminant, in clams at least.     

Polycyclic aromatic hydrocarbons in surface seawater and in indigenous mussels (Mytilus galloprovincialis) from coastal areas of the Saronikos Gulf (Greece)

Polycyclic aromatic hydrocarbons (PAHs) were identified and measured in surface seawater and in the tissues (gills and mantle) of indigenous black mussels, Mytilus galloprovincialis, collected from three coastal sites of Saronikos Gulf (Greece), a gulf that exhibits high levels of pollution. The total PAHs measured by spectrofluorometry in the surface seawater were found in the range of 425–459 ng L⁻¹ at the most polluted sites 1 and 2 (Elefsis Bay–Salamis Island) and in the range of 103–124 ng L⁻¹ at site 3 (Aegina Island). PAHs' sources in seawater were identified by application of specific PAH ratios, such as phenanthrene/anthracene and fluoranthene/pyrene. Levels of PAHs in soft tissues (gills and mantle) of indigenous mussels were much higher than those reported for seawater. Total PAH concentrations in mantle tissues were in the range of 1300–1800 ng g⁻¹ dry weight (dw) tissue at sites 1 and 2 and approximately 380 ng g⁻¹ dw at site 3. In gill tissues total PAH concentrations were in the range of 1480–2400 ng g⁻¹ dw at sites 1 and 2 and approximately 430 ng g⁻¹ dw at site 3. PAHs composition was dominated by two-, three- and four-ring compounds in seawater, where 17 different PAH compounds were identified and measured in mussel tissues. Mussels can be used as sentinel organisms to monitoring PAHs' contamination, since they concentrate PAHs from the surrounding water media and therefore making the chemical analysis simpler and less prone to error than that for water. In surface seawater possible weathering and photodegradation due to hot climates contribute to reduced PAHs concentrations.     

Preliminary evaluation of the toxic effects of the antifouling biocide Sea-Nine 211™ in the soft coral Sarcophyton cf. glaucum (Octocorallia,Alcyonacea) based on PAM fluorometry and biomarkers

Sea-Nine 211™ is a new biocide specifically formulated for antifouling paints and being considered to have a low environmental impact. Even with a short environmental half-life, this compound can cause toxic effects on marine organisms. This study used PAM fluorometry and biomarkers of oxidative stress (GST, CAT and LPO) to monitor potential toxic effects of Sea-Nine 211™ on fragments of the soft coral Sarcophyton cf. glaucum. After exposure to concentrations of 1–100 μg l⁻¹ for 72 h, CAT activity was inhibited under the two highest concentrations, being in accordance with the activity of GST. LPO activity (as TBARS) and photosynthetic efficiency of endosymbiotic zooxanthellae were not significantly affected. These results show that PAM fluorometry alone cannot detect the full effects of Sea-Nine 211™ on Sarcophyton cf. glaucum and should be used together with other biomarkers. This holobiont driven approach to evaluate chemical toxicity in photosynthetic corals is therefore recommended for biocides which are not photosystem II inhibitors.     

Connectivity clues from short-term variability in settlement and geochemical tags of mytilid mussels

The use of geochemical tags in calcified structures of fish and invertebrates is an exciting tool for investigating larval population connectivity. Tag evaluation over relatively short intervals (weeks) may detect environmental and ecological variability at a temporal scale highly relevant to larval transport and settlement. We collected newly settled mussels (Mytilus californianus and M. galloprovincialis) weekly during winter/spring of 2002 along the coast of San Diego, CA, USA, at sites on the exposed coast (SIO) and in a protected coastal bay (HI), to investigate temporal patterns of geochemical tags in mussel shells. Analyses of post-settlement shell via LA-ICP-MS revealed statistically significant temporal variability for all elements we examined (Mg, Mn, Cu, Sr, Cd, Ba, Pb and U). Despite this, our ability to distinguish multielemental signatures between sites was largely conserved. Throughout our 13-week study, SIO and HI mussels could be chemically distinguished from one another in 78–87% of all cases. Settlement varied between 2 and 27 settlers gram-byssus^?1 week^?1 at SIO and HI, and both sites were characterized by 2–3 weeks with “high” settlement. Geochemical tags recorded in early larval shell of newly settled mussels differed between “high” and “low” settlement weeks at both sites (MANOVA), driven by Mg and Sr at SIO (p = 0.013) and Sr, Cd, Ba and Pb at HI (p < 0.001). These data imply that shifts in larval sources or transport corridors were responsible for observed settlement variation, rather than increased larval production. In particular, increased settlement at HI was observed concurrent with the appearance of geochemical tags (e.g., elevated Cd), suggesting that those larvae were retained in upwelled water near the mouth of the bay. Such shifts may reflect short-term changes in connectivity among sites due to altered transport corridors, and influence the demography of local populations.     

Enhancement of polycyclic aromatic hydrocarbons in estuarine invertebrates by surface runoff at a decommissioned military fuel depot

Accumulation of polycyclic aromatic hydrocarbons (PAHs) was determined in blue mussels (Mytilus spp.) and shore crabs (Hemigrapsus sp.) at a recently closed military fuel depot in central San Francisco Bay, California. In April 1996, during a period of above average precipitation, specimens were collected at the depot, near the depot, and at sites 10 and 20 km south of the depot. Four weeks after the rains ended, blue mussels were again collected at the depot, and at two additional sites in the central Bay region. In April, total PAHs in mussels from the depot were significantly higher only than that in mussels collected 20 km from the depot; however, seven specific, substituted PAHs were higher at the depot than at all other sites. In June, only two of the 38 PAHs common in mussels in April were detected at the depot; these concentrations were comparable to ambient concentrations in mussels at the Bay. It seemed that bioavailability of PAHs at the depot was enhanced by rainfall, probably due to the mobilization of PAHs via groundwater into the Bay. Concentrations in mussels from chronically contaminated sites were about five times higher than mussels collected from the depot. Low PAH concentrations were detected in shore crabs near the depot, and the highest levels were not associated with the depot. Observed PAH concentrations are discussed in relation to upper trophic organisms.     

Trace metal content in sediments and autochthonous intertidal organisms from two adjacent bays near Ushuaia,Beagle Channel (Argentina)

The aim of this work was to monitor levels of Cd, Cu, Pb, Zn and Fe in sediments, mussels (Mytilus edulis chilensis) and limpets (Nacella magellanica) from the Industrial zone (IZ); fuel dock (FD) and Ushuaia Peninsula (UP) on the Beagle Channel. In sediments, seasonal variations showed high values of Cu and Pb in spring and Zn in autumn. Comparing among sites, Cd concentration was superior in UP (2.07 μg/g); while Pb was maximum in FD (41.00 μg/g). In mussels, a higher bioaccumulation in winter was found. Mussels from UP showed the highest bioaccumulation of Cu (5.95 μg/g) and those from FD presented the highest of Zn (170.15 μg/g). A seasonal trend was not found for limpets, while differences among sites were observed for Cd being the highest at IZ (3.02 μg/g). Although pollution level found was low, anthropic activities at the studied sites could result in deterioration, further monitoring is recommended.     

Polycyclic aromatic hydrocarbon (PAH) burden of mussels (Mytilus sp.) in different marine environments in relation with sediment PAH contamination, and bioavailability

Sediments and mussels (Mytilus edulis, Mytilus galloprovincialis) were sampled in different European coastal environments (Germany, France, Spain) and analysed for their polycyclic aromatic hydrocarbon (PAH) content by Gas Chromatography/Mass Spectrometry (GC-MS). Bioaccumulation factors of individual compounds from the sediment were calculated and discussed according to the compound solubility. The mussels showed different accumulation patterns according to the pollution source they were exposed to (dissolved fraction of PAHs, particulate fraction, petroleum present in the water column). The exposure source also depends on the geographical location of the mussels. In the Mediterranean Sea, the bivalves were mainly exposed to the dissolved fraction of PAHs, while in the Baltic Sea and in the Atlantic Ocean, the PAHs associated to the particles were significant sources.     

Metal pollution assessment of the marine environment by determination of metal-binding proteins in Mytilus sp.

Adult mussels (Mytilus galloprovincialis) (shell length 5–7 cm) exposed to elevated concentrations of Cd (0.1–1.3 mg l⁻¹), in a flowing seawater aquarium under laboratory conditions, responded by elevated synthesis of metal-binding proteins in a digestive gland. Response depended on the Cd concentration level as well as on duration of the exposure time. Results obtained show that inducible proteins (isolated by gel-filtration column chromatography and characterized by electrochemical and spectrophotometric methods) should be considered metallothionein-like proteins.The induced metal-binding proteins of mussels in specific organs or tissues are considered to be a potential biological indicator of metal pollution, taking into account specific qualities of the mussel response to different metals. The advantages of the analytical methods applied (gel-chromatography and polarography) and the relevance of the laboratory data when transferred to environmental conditions are discussed. Previously reported results dealing with metal-binding proteins of Mytilus species are also summarized.     

The effect of shore location on biomarker expression in wild Mytilus spp. and its comparison with long line cultivated mussels

Biomarkers are a common tool in the assessment of potential effects of contaminants in aquatic organisms. In order to identify the effects of anthropogenic pollution it is essential to identify background levels and to know the range of natural variability in the biomarker response. In this study, we examined various biomarkers of stress (glutathione S-transferase and metallothionein), damage (lipid peroxidation (LPO) and DNA damage (DNA)) and reproduction (vitellin-like proteins) in marine mussels (Mytilus spp.) from four locations along a vertical transect from high to low shore and compared them with cultivated long line mussels. High shore and cultivated mussels showed significantly higher LPO and DNA damage expression than the low shore mussels indicating a level of oxidative stress resulting from mussel location. Significant effects in physiological endpoints were also found. This study highlights the need to consider the diversity of natural environmental stress factors when using biomarkers in environmental assessment.     

Expression and activity of glutathione S-transferases and catalase in the shrimp Litopenaeus vannamei inoculated with a toxic Microcystis aeruginosa strain

Microcystin (MC) produced during cyanobacteria blooms is notably toxic to human and wildlife. Conjugation with reduced glutathione (GSH) by glutathione S-transferase (GST) and the antioxidant enzymes defenses (e.g. catalase, CAT) are important biochemical defense mechanisms against MCs toxicity. We investigated the enzymatic activity of CAT and GST and the gene expression levels of CAT and eight GST isoforms in the hepatopancreas of the globally farmed shrimp Litopenaeus vannamei 48-h after injection with a sub-lethal dose of 100 μg kg⁻¹ of a toxic Microcystis aeruginosa extract. MCs caused up-regulation for GST Ω, μ and a MAPEG isoform, by 12-, 2.8- and 1.8-fold, respectively, and increases in the total GST enzyme activity and CAT enzyme activity. The study points to the importance of further characterization for the L. vannamei GST isoforms and GST/CAT post-translational regulation processes to better understand the key mechanisms involved in the shrimp’s defense against MC exposure.     

Phase I and II biotransformation enzymes and polycyclic aromatic hydrocarbons in the Mediterranean mussel (Mytilus galloprovincialis,Lamarck, 1819) collected in front of an oil refinery

The aim of the present study was to investigate the responses of phase I and II biotransformation enzymes and levels of PAHs in the Mediterranean mussel (Mytilus galloprovincialis, Lamarck, 1819) collected from three sites at different distance from an oil refinery. Phase I enzyme activities as NAD(P)H-cyt c red, NADH ferry red, B(a)PMO and phase II as UDPGT, GST were measured in digestive gland while 16 PAHs (US-EPA) in whole soft tissue. An added value to the data obtained in the present study rely on the RDA analysis which showed close correlations between PAHs levels and phase I enzyme activities in mussels collected in front of the refinery. And again a significant spatial correlation between B(a)P levels and NADPH-cyt c red activities was observed using linear models. No differences among sites for B(a)PMO and phase II GST activities were observed, while the application of UDPGT as biomarkers requires further investigation.     

Algal concentration profiles above mussel beds

The distribution of phytoplankton was measured in vertical profiles at three stations in Limfjorden (Denmark) within and above Mytilus edulis beds (−5 to 200 cm above the bed surface) and related to the filtration capacity of the mussels and hydrodynamic conditions. The density of phytoplankton was 2690 to 10 600 cells cm⁻³ at 200 cm above the bottom (∼1.4 to 3.2 mg Chl-a m⁻³), declining towards the bottom to about 1000 cells cm⁻³ at two of the stations. This is a threshold of minimum algal concentration below which mussels are reported to stop feeding. The results of the present study further demonstrate that the near-bed phytoplankton concentrations observed in the field often remained below the concentration level at which mussels reduce their maximal filtration activity.     

Implications from a field study regarding the relationship between polycyclic aromatic hydrocarbons and glutathione S-transferase activity in mussels

An aluminium smelter on the west coast of Scotland discharges an aqueous effluent containing polycyclic aromatic hydrocarbons (PAHs) at the head of Loch Leven. The loch also supports two mussel (Mytilus edulis) farms. Data are presented on burdens of PAHs in the soft tissues of mussels and the effect of these contaminants on glutathione S-transferase (GST) activity in mussel hepatopancreas. GST activity is shown to be correlated with total PAH burden and also with the concentrations of certain individual PAHs. These field data show that high molecular weight PAHs are closely correlated to GST activity, whereas low molecular weight PAHs are not. This suggests that 5- and 6-ring PAHs have a more pronounced role than 2- to 4-ring compounds in inducing GST activity in mussels from Loch Leven. It is proposed that it may be more appropriate to link GST activity with 5- and 6-ring compounds only, rather than with the total PAH burden.     

Pyrene exposure influences the craniofacial cartilage development of Sebastiscus marmoratus embryos

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental contaminants, which are known carcinogens and teratogens. However, the toxicity of PAHs during skeletal development and the mechanism involved are not completely clear. In the present study, rockfish (Sebastiscus marmoratus) embryos were exposed to pyrene (Pyr) for 7 days at 0.5, 5 and 50 nM which resulted in craniofacial skeleton deformities. Pyr exposure for 6 days reduced the expression of PCNA, Col2a1 and Sox9 in the craniofacial skeleton revealed using in situ hybridization. These results suggest that Pyr exposure impairs skeleton development via disrupting the proliferation of the chondrocytes. At the same time, Pyr exposure reduced the expression of lox1 and inhibited the activity of lysyl oxidase, which is the key enzyme controlling the collagen cross-linking, and which might therefore have been one of the reasons for the deformative Meckel's cartilage (lower jaw).     

Physical drivers of phytoplankton production in the southern Benguela upwelling system

Investigations of primary production (PP) were undertaken in the southern Benguela ecosystem during two research surveys in October 2006 and May 2007. Significant differences in environmental conditions, as well as biomass and PP, were observed between October and May. During October, integrated biomass and PP were significantly higher, ranging from 20.43 to 355.01 mg m⁻², and 0.71 to 6.98 g C m⁻² d⁻¹, respectively, than in May, where the range was 47.92–141.79 mg m⁻², and 0.70–3.35 g C m⁻² d⁻¹, respectively. Distribution patterns indicated low biomass and PP in newly upwelled water along the coast, higher biomass and PP in the mid-shelf region, while lower values were observed at and beyond the shelf edge. Latitudinal variations showed consistently higher biomass and PP in the St. Helena Bay region compared to biomass and PP south of Cape Town. During both surveys, phytoplankton communities were comprised primarily of diatoms and small flagellates, with no significant differences. Phytoplankton adaptation to environmental variability was characterised by increased P_m^B and E_k under elevated temperatures and irradiance, while no clear relationships were evident for α^B. Generalised Additive Models (GAMs) showed that photosynthetic parameters were all significant predictors of photosynthesis rates (P_z), with P_m^B being the most important, accounting for 36.97% of the deviance in P_z. However, biomass levels and environmental conditions exerted a much greater influence on P_z, with irradiance explaining the largest proportion (68.24%) of the deviance. Multiple predictor GAMs revealed that 96.26% of the deviance in P_z could be explained by a model which included nitrate, chlorophyll a, and irradiance.     

Abundance of small individuals influences the effectiveness of processing techniques for deep-sea nematodes

Nematodes are the most abundant metazoans of deep-sea benthic communities, but knowledge of their distribution is limited relative to larger organisms. Whilst some aspects of nematode processing techniques, such as extraction, have been extensively studied, other key elements have attracted little attention. We compared the effect of (1) mesh size (63, 45, and 32 μm) on estimates of nematode abundance, biomass, and body size, and (2) microscope magnification (50× and 100×) on estimates of nematode abundance at bathyal sites (250–3100 m water depth) on the Challenger Plateau and Chatham Rise, south-west Pacific Ocean. Variation in the effectiveness of these techniques was assessed in relation to nematode body size and environmental parameters (water depth, sediment organic matter content, %silt/clay, and chloroplastic pigments). The 63-μm mesh retained a relatively low proportion of total nematode abundance (mean±SD=55±9%), but most of nematode biomass (90±4%). The proportion of nematode abundance retained on the 45-μm mesh in surface (0–1 cm) and subsurface (1–5 cm) sediment was significantly correlated (P<0.01) with %silt/clay (R²=0.39) and chloroplastic pigments (R²=0.29), respectively. Variation in median nematode body weight showed similar trends, but relationships between mean nematode body weight and environmental parameters were either relatively weak (subsurface sediment) or not significant (surface sediment). Using a low magnification led to significantly lower (on average by 43%) nematode abundance estimates relative to high magnification (P<0.001), and the magnitude of this difference was significantly correlated (P<0.05) with total nematode abundance (R²_p=0.53) and the number of small (≤250 μm length) individuals (R²_p=0.05). Our results suggest that organic matter input and sediment characteristics influence the abundance of small nematodes in bathyal communities. The abundance of small individuals can, in turn, influence abundance estimates obtained using different mesh sizes and microscope magnifications.     

Analysis of a PAH-degrading bacterial population in subsurface sediments on the Mid-Atlantic Ridge

Little is known about the types and concentrations of polycyclic aromatic hydrocarbons (PAHs) existing in the deep-sea subsurface environment, which is believed to be cold, oligothrophic and of high static pressure. PAHs in the upper layers of the water column are unavoidably subjected to degradation while they are deposited to the sea floor and become embedded in the deep-sea sediment. In this report, a high concentration of PAHs was discovered in the sediment 2.7 m beneath the bottom surface at a water depth of 3962 m on the Mid-Atlantic Ridge (MAR). The total concentration of PAHs was 445 ng (g dry wt sediment)^-1. Among the seven detected PAHs, the concentrations of phenanthrene (222 ng g^-1) and fluorene (79 ng g^-1) were relatively high. In addition, PAH-degrading bacteria were found within the sediments. As in a previously detected site on the MAR, in the PAH-enriched region of this site, a bacterium of the genus Cycloclasticus was found to be the predominant isolate detected by PCR-DGGE analysis. In addition, bacteria of the Halomonas, Marinobacter, Alcanivorax, Thalassospira and Maricaulis genera, were also included in the PAH-degrading community. In summary, a high concentration of PAHs was detected in the subsurface of the deep-sea sediment, and once again, the Cycloclasticus bacterium was confirmed to be a ubiquitous marine PAH degrader even in the subsurface marine environment. Considering the abundance of PAHs therein, biodegradation is thus thought to be inactive, probably because of the low temperature, limited oxygen and/or limited nutrients.