Uptake and depuration of PAHs and chlorinated pesticides by semi-permeable membrane devices (SPMDs) and green-lipped mussels (Perna viridis)

Semi-batch seawater experiments were conducted to follow the uptake and release of selected PAHs (anthracene, fluorathene, pyrene and B[a]P) and organochlorine pesticides (-HCH, aldrin, dieldrin, p,p′-DDT) in semi-permeable membrane devices (SPMDs) and green-lipped mussels (Perna viridis). Mathematical models were applied to describe the uptake and elimination curves of the contaminants for SPMDs, and kinetic parameters, such as uptake rate constants, and equilibrium triolein/water partitioning coefficients were calculated. SPMD data showed a good fit to estimate rate constant and partition coefficient equations, but only those contaminants which partitioned mainly in the dissolved phase (-HCH and dieldrin) were well explained for mussels. Poor conformity of the other contaminants indicated mussels uptake by routes other than diffusion, such as ingestion of algae. An apparent equilibrium state was only noted for -HCH in mussels. Aldrin was not detected in mussels in the first few days of exposure, indicating potential metabolism of this compound. B[a]P was not detected in the triolein of SPMDs, which suggests that the membrane may act as a reservoir. Loss of spiked B[a]P from the triolein was evident in a depuration experiment, which may indicate transfer to the membrane. Rate constants for mussels were higher than those for SPMDs, but the reverse was true for partition coefficients. Overall, mussels and SPMDs had similar uptake rates for all compounds in this study, excluding p,p′-DDT and dieldrin. Contaminant elimination took place more rapidly in mussels, implying that SPMDs are better candidates for detecting episodic discharge of organic contaminants.     

Biofouling confounds the uptake of trace organic contaminants by semi-permeable membrane devices (SPMDs)

The outer layers of layflat, low density polyethylene plastic tubing (the principal component of semi-permeable membrane devices, SPMDs) were biofouled at a clean site in Hong Kong coastal waters for periods of 1–4 weeks. Following pre-fouling, triolein was added to the SPMDs and, along with control (unfouled) devices, they were exposed to a range of organochlorine pesticides (-HCH, aldrin, p,p^′-DDT) and PAHs (anthracene, fluoranthene and benzo(a)pyrene) under laboratory conditions. Results showed that the uptake of contaminants by SPMDs was severely reduced by as much as 50% under fouling conditions in comparison to unfouled controls. The ultimate utility of SPMDs as passive monitors is thus reduced, although alternative measures, such as the use of permeability reference compounds may compensate, and allow for realistic evaluations of dissolved environmental concentrations in aquatic environments. However, due to the complexities involved in such procedures––especially as they need to be conducted on a case-by-case basis––the utility of SPMDs appears to be limited for estimates of bioavailability unless necessary calibrations are undertaken within each environment that the sampler is used.     

Uptake and depuration of PAHs and chlorinated pesticides by semi-permeable membrane devices (SPMDs) and green-lipped mussels (Perna viridis)

Semi-batch seawater experiments were conducted to follow the uptake and release of selected PAHs (anthracene, fluorathene, pyrene and B[a]P) and organochlorine pesticides (α-HCH, aldrin, dieldrin, p,p′-DDT) in semi-permeable membrane devices (SPMDs) and green-lipped mussels (Perna viridis). Mathematical models were applied to describe the uptake and elimination curves of the contaminants for SPMDs, and kinetic parameters, such as uptake rate constants, and equilibrium triolein/water partitioning coefficients were calculated. SPMD data showed a good fit to estimate rate constant and partition coefficient equations, but only those contaminants which partitioned mainly in the dissolved phase (α-HCH and dieldrin) were well explained for mussels. Poor conformity of the other contaminants indicated mussels uptake by routes other than diffusion, such as ingestion of algae. An apparent equilibrium state was only noted for α-HCH in mussels. Aldrin was not detected in mussels in the first few days of exposure, indicating potential metabolism of this compound. B[a]P was not detected in the triolein of SPMDs, which suggests that the membrane may act as a reservoir. Loss of spiked B[a]P from the triolein was evident in a depuration experiment, which may indicate transfer to the membrane. Rate constants for mussels were higher than those for SPMDs, but the reverse was true for partition coefficients. Overall, mussels and SPMDs had similar uptake rates for all compounds in this study, excluding p,p′-DDT and dieldrin. Contaminant elimination took place more rapidly in mussels, implying that SPMDs are better candidates for detecting episodic discharge of organic contaminants.     

Comparison of mussels and semi-permeable membrane devices as intertidal monitors of polycyclic aromatic hydrocarbons at oil spill sites

Side-by-side comparisons of polycyclic aromatic hydrocarbon (PAH) concentrations in resident blue mussels (Mytilus trossulus) and in semi-permeable membrane devices (SPMDs) were made at four sites in Prince William Sound, Alaska. SPMDs were deployed for approximately 30 days on the surface of the beach sediment at three tidal elevations on each shore and in 0.5 m deep open pits in the middle intertidal zone. Total PAH (TPAH) concentrations in mussels and in SPMDs were correlated, but the PAH compositions were different. The lower molecular weight PAH were relatively more abundant in the SPMDs than in the mussels at oiled and HA sites. TPAH concentrations in SPMDs deployed in pits and mussels collected adjacent to those pits at oiled sites were higher than in SPMDs and mussels from non-pitted SPMD locations approximately 3-15 m from the pits. Pitting released buried oil making its PAH bioavailable. SPMDs deployed in the supratidal zone (+4.0 m tidal elevation) were exposed to atmospheric contaminants for a large fraction of the deployment time and accumulated primarily pyrogenic (combustion-sourced) PAH from the atmosphere. The SPMD strips supplied by the manufacturer contained significant amounts (approximately 125 ng/strip) of primarily alkylated 2-3 ring PAH. These blank levels make SPMDs unsuitable for shoreline assessments when environmental PAH concentrations are low. Consequently, where available, mussels are recommended for use in assessments of the bioavailability of buried oil residues sequestered in intertidal sediments following an oil spill. Mussels are the preferred monitoring tool when the assessments involve food-chain effects. At locations where the absence of mussels necessitates the use of SPMDs or other passive sampling devices, their limitations need to be carefully considered in the interpretation of results.     

Protective effect of phospholipid hydroperoxide glutathione peroxidase (PHGPx) against lipid peroxidation in mussels Perna perna exposed to different metals

Levels of antioxidant defenses and lipid peroxidation were evaluated in mussels exposed to lead (200 mg/l), iron (500 microg/l), cadmium (200 microg/l) and copper (40 microg/l), for 12, 24, 72 and 120 h. Glutathione S-transferase (GST) activity was unchanged with all treatments. Catalase (CAT) increased after 120 h of exposure to all metals. Mussels exposed to Cd for 12 h, and to Cu and Fe for 120 h had increased lipid peroxidation, which might be associated to decreased levels of reduced glutathione (GSH) and glutathione peroxidase (GPx) activity. Pb exposure caused GSH depletion after 12 h and increased GPx activity after 120 h. Negative correlations were observed between the enzyme phospholipid hydroperoxide glutathione peroxidase (PHGPx) and malonaldehyde (MDA) levels after Fe and Cu exposure, indicating a protective role of PHGPx against lipid peroxidation, and suggesting the use of this enzyme as a new potential biomarker of toxicity associated with contaminant exposure in mussels.     

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

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

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

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

Bioenergetic responses in green lipped mussels (Perna viridis) as indicators of pollution stress in Xiamen coastal waters, China

Samples of green lipped mussels (Perna viridis) were collected from three sites (Huoshao, I’Maluan and Tong’an) around Xiamen coastal waters, where levels of various trace organic pollutants have been studied. Samples were also collected at a “cleaner” reference site near Dongshan Island for comparison. Clearance rate, absorption efficiency and oxygen consumption of the mussels were measured under controlled laboratory condition; organic pollutants in their tissues were also analyzed. Scope for growth (SFG) was employed as a general biomarker to evaluate the stress of pollutants. Specimens from Tong’an site had the lowest SFG values (1.14 J/h/g); while specimens from the I’Maluan site and Huoshao site had SFG values of 5.01 and 6.72 J/h/g, respectively. Specimens from Dongshan (reference site) had a relatively high SFG value of 10.96 J/h/g. There was a significant negative correlation between the SFG of the different populations of green lipped mussels and the concentration of DDT in their tissues.     

Competitive sorption of persistent organic pollutants onto microplastics in the marine environment

Plastics are known to sorb persistent organic pollutants from seawater. However, studies to quantify sorption rates have only considered the affinity of chemicals in isolation, unlike the conditions in the environment where contaminants are present as complex mixtures. Here we examine whether phenanthrene and 4,4′-DDT, in a mixture, compete for sorption sites onto PVC with no added additives (unplasticised PVC or uPVC) and Ultra-High Molecular Weight polyethylene. Interactions were investigated by exposing particles of uPVC and UHMW PE to mixtures of 3H and 14C radiolabelled Phe and DDT. Changes in sorption capacity were modelled by applying a Freundlich binding sorption isotherms. An Extended Langmuir Model and an Interaction Factor Model were also applied to predict equilibrium concentrations of pollutants onto plastic. This study showed that in a bi-solute system, DDT exhibited no significantly different sorption behaviour than in single solute systems. However, DDT did appear to interfere with the sorption of Phe onto plastic, indicating an antagonistic effect.     

Evaluation of the mussel Perna perna as a biomonitor of polycyclic aromatic hydrocarbon (PAH) exposure and effects

The mussel Perna perna was used to monitor and assess areas suspected of oil contamination by transplanting animals from unimpacted to impacted sites and vice versa. The deployment lasted for three months during which total PAH were measured in seawater samples collected every 15 days. PAH concentrations were determined in mussel tissue monthly. The lysosomal neutral red assay and the condition index were used as biomarkers of the adverse biological effects. The PAH concentration in mussels at unimpacted sites, prior to deployment was ca. 100 microg kg(-1) but increased to 300 microg kg(-1) after three months at the contaminated site. Mussels transplanted to the uncontaminated site showed a decrease from 380 to 80 microg kg(-1). Significant changes in lysosomal stability occurred within one month of the beginning of the transposition; in contrast condition indexes were not correlated with the observed changes in PAH concentration.     

Occurrence of persistent organic pollutants in marine fish from the Natuna Island,South China Sea

Five marine fish species were collected from the Natuna Island, South China Sea to investigate the occurrence of polybrominated diphenyl ethers (PBDEs), polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane and its metabolites (DDTs). Concentrations of PBDEs, PCBs, and DDTs in marine fish ranged from 2.85 to 7.82, 14.3 to 48.1, and 7.99 to 40.3 ng/g lipid weight, respectively. Higher concentrations of PBDEs, PCBs, and DDTs were observed in Snakefish (Trachinocephalus myops), which might be attributed to their different feeding and living habits. PCBs were the predominant POPs in all marine fish, followed by DDTs and PBDEs. BDE 47 and PCB 153 were the predominant congener of PBDEs and PCBs, respectively. Compositional distribution of DDTs indicated the possible presence of fresh input sources around the Natuna Island. The ratios of o,p′-DDT/p,p′-DDT being less than 1 in fish samples suggested that DDT contributions from dicofol seemed considerably low. New input sources of DDT in South China Sea are worth further research.     

Relationships between tissue concentrations of chlorinated hydrocarbons (polychlorinated biphenyls and chlorinated pesticides) and antioxidative responses of marine mussels,Perna viridis

Marine mussels, Perna viridis, were transplanted from a reference site to various polluted sites around Hong Kong. After 30 d of exposure, antioxidative responses in the gills and hepatopancreas and tissue concentrations of chlorinated hydrocarbons [polychlorinated biphenyls (PCBs) and chlorinated pesticides (CPs)] were determined for individual mussels. Glutathione S transferase (GST) and glutathione (GSH) were positively correlated with tissue PCB concentrations. Only one of the enzymatic antioxidants, glutathione peroxidase (GPx), showed significant response to tissue PCB. No significant correlation was found between tissue concentrations of chlorinated hydrocarbons and other enzymatic antioxidants (superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR) and NADPH DT-diaphorase (DT-d). Oxidative stress, measured as thiobarbituric acid reactive substances, was correlated with chlorinated pesticide concentrations in tissues. This study demonstrated a correlation between GST/GSH and chlorinated hydrocarbons. The apparent lack of correlation between trace organic pollutants and some of the enzymatic antioxidants may be due to the inhibitory effects caused by these chemicals. The above results suggest that more investigations are needed before these enzymes can be used as biomarkers.     

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.     

Field validation of antioxidant enzyme biomarkers in mussels (Perna viridis) and clams (Ruditapes philippinarum) transplanted in Hong Kong coastal waters

Green-lipped mussels, Perna viridis, and Manila clams, Ruditapes philippinarum were sourced from “clean” sites in the Hong Kong region, depurated in a laboratory using uncontaminated filtered seawater for 8 days, and transplanted to a suspected gradient of chemically polluted sites in Hong Kong. After 14- and 28-days of field exposure, several antioxidant parameters including glutathione S transferase (GST), catalase (CAT), glutathione peroxidase (GPx), and glutathione (GSH) were quantified in gill and hepatopancreas tissues. Whole body tissue concentrations of polycyclic aromatic hydrocarbons (PAHs), petroleum hydrocarbons (PHCs), polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCs) were determined in pooled site samples. Chemical analyses indicated that: (a) clams had higher levels of PAHs, PHCs, DDTs and PCBs, whereas mussels had higher hexachlorocyclohexane (HCHs) and there was no difference between species for dieldrin and remaining OCs; (b) Kat O should not be continued as a “clean” reference site for Hong Kong, because of the levels of contaminants measured and (c) PAH concentrations in the current survey were similar to those previously measured. Toxicological conclusions were: (a) antioxidant responses were different between species; (b) CAT and GST have highest utility in clams for field use in Hong Kong, whereas CAT in both gill and hepatopancreas tissue showed most potential in mussels; (c) significant induction of antioxidant responses over day 0 (excluding GPx in both tissues, and GST in mussel hepatic tissue); (d) groups of contaminants do not consistently induce antioxidant responses and (e) organochlorines and PCBs correlated significantly with CAT and GST in clam hepatopancreas and with CAT in mussel gill and hepatic tissue. Multivariate statistical techniques indicated little relationship between the site patterns for antioxidant responses and the contaminant gradients identified in body burden analysis.     

Correlations between metal uptake in the soft tissue of Perna perna and gill filament pathology after exposure to mercury

The accumulation of metal in soft tissues, filtration rate and gill filament morphology are correlated in the southern African rock mussel, Perna perna, during exposure to mercury (24 days) and recovery (24 days). The amount of Hg in soft tissues increased from 0.13 to 87.5 μg/g dry weight after 24 days exposure, and declined to 13 μg/g during recovery. Mean filtration rate fell from 3979 to 1818 ml/h/g dry weight by day 2, but recovered slightly through days 4 and 8 (3037 ml/h/g), with a higher average rate (5030 ml/h/g) being maintained over the 24–48 days recovery period. The initial decline in filtration coincided with epithelial cell deterioration presented as interstitial oedema, neural and epithelial cell degeneration and reduced ciliation. Between days 8 and 24, cilia regenerated and there was a general improvement in cell morphology. Gill filament morphology returned to near normal during the metal-free recovery period. The usefulness of P. perna as an indicator of pollution is discussed.     

Polar organic micropollutants in the coastal environment of different marine systems

Polar anthropogenic organic micropollutants are frequently detected in freshwater and discharged on large scale into marine systems. In this work the results of 153 samples collected from the shorelines of the Baltic Sea (Germany), Northern Adriatic Sea (Italy), Aegean Sea and Dardanelles (Greece & Turkey), San Francisco Bay (USA), Pacific Ocean (USA), Mediterranean Sea (Israel), and Balearic Sea (Spain) are presented. The samples were analyzed for various classes of micropollutants such as pharmaceuticals, corrosion inhibitors, biocides, and stimulants. Caffeine, paraxanthine, theobromine, tolyltriazole, 1H-benzotriazole, and atrazine were detected in >50% of all samples. The detection frequencies of carbamazepine, iopamidol, diuron, sulfamethoxazole, paracetamol, theophylline, and atenolol were between 20% and 32%. As caffeine is linked to untreated wastewater, the widespread occurrence of raw sewage in marine environments and thus potentially elevated nutrient concentrations and risk for the presence of wastewater-related pathogens is remarkable.     

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.