Lynda Webster

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.

Long-term Monitoring of Polycyclic Aromatic Hydrocarbons in Mussels (Mytilus edulis) Following the Braer Oil Spill†

On January 5, 1993, 84700 t of Norwegian Gullfaks crude oil was released into the coastal region of south Shetland when the tanker MV Braer grounded at Garths Ness. A Fisheries Exclusion Zone was designated under the Food and Environment Protection Act 1985 (FEPA), prohibiting the taking or harvesting of fish or shellfish within the Zone so as to prevent contaminated products reaching the marketplace. The criteria set for lifting of the Order were that the particular species must be free from any petrogenic taint and the concentration of polycyclic aromatic hydrocarbons (PAHs) must be within the range for reference samples. Between April 1993 and February 1995 the Order was progressively lifted for wild fish, salmon, crustacea, excluding Nephrops norvegicus (Norway lobster), and molluscs, with the exception of mussels. As part of the monitoring exercise, mussels from a reference site were transplanted in June 1995 to three sites within the Zone, where they were suspended in plastic mesh boxes from rafts to a depth of 5 m. Samples were collected at regular intervals over the following 12 months and the concentration and composition of PAHs were determined by gas chromatography with mass spectrometric detection. The total measured PAH concentration at the control site increased from 13.7 to 66.1 ng g–1 wet mass of tissue between June 1995 and February 1996. This trend was reversed by July 1996 when the PAH concentration was 12.8 ng g–1. The mean across the year for the control site was 24.0 ng g–1 (SE = 8.9 ng g–1, n = 6). A similar seasonal trend in PAH concentration over the year was observed at all sites within the Zone, but the PAH concentration was consistently greater at these sites, reaching a maximum concentration of 316 ng g–1 in February 1996. Although no taint was detected in any of the mussels, these results meant that it was not possible to lift the Prohibition Order for mussels. Further monitoring at three sites outwith the Zone and three sites within the Zone is under way together with investigations into the specific source of the PAHs.

Passive sampling: partition coefficients for a silicone rubber reference phase

Silicone rubber sheeting can be used as a passive sampling device for hydrophobic organic contaminants in the environment to determine the available concentrations in water and sediments. Reliable sampler-water partition coefficients are required to determine the sampling rates and the dissolved contaminant concentrations in water and in sediment pore water. Log partition coefficients (logK(sr,w)) for silicone rubber-water have been estimated for 32 polycyclic aromatic hydrocarbons (PAHs), 2 deuterated PAH analogues and 32 chlorobiphenyls (CBs) using the cosolvent method, with methanol as cosolvent. Strong linear relationships were found with literature values for the corresponding log octanol-water partition coefficients (logK(ow)) for both CBs and PAHs, confirming that partitioning into the silicone rubber is strongly determined by the hydrophobicity of the compounds, which suggests logK(ow) is a good predictor of logK(sr,w) and that absorption is the main mechanism for accumulation of analytes into the silicone rubber polymer.

The authentication of olive oil on the basis of hydrocarbon concentration and composition

Samples of virgin olive oil and refined olive oil were analysed for n-alkanes by gas chromatography with flame ionisation detection to determine if the pattern and composition were oil specific and, therefore, if the hydrocarbon patterns could be used as determinants for assessing adulteration of olive oil. The carbon number profile of the extra virgin olive oil was unique inasmuch as the odd numbered predominance was not limited to two or three n-alkanes but started at tricosane (nC23) and continued to tritriacontane (nC33). The olive oil n-alkane data was added to an existing database that included rapeseed, safflower, sunflower, corn, palm, palm kernel, coconut, groundnut and soyabean oils and analysed by principal component analysis (PCA). Olive oil could clearly be differentiated from the other vegetable oils. PCA also allowed for the distinction of olive oils from different geographical regions. Authentic extra virgin olive oil was adulterated with various amounts of either crude sunflower or crude rapeseed oil, which resulted in adulteration levels between 0.5 and 11% w/w. Using the carbon number profiles alone it was possible to determine adulteration of the extra virgin olive oil with as little as 2.6% crude rapeseed oil or crude sunflower oil. Analysis of the n-alkane pattern by PCA made it possible to identify adulterants at levels as low as 0.5% w/w.

The polycyclic aromatic hydrocarbon and geochemical biomarker composition of sediments from voes and coastal areas in the Shetland and Orkney Islands

Marine sediments from coastal areas and voes in the Shetland and Orkney Islands were analysed for parent and branched 2- to 6-ring polycyclic aromatic hydrocarbons (PAHs) and geochemical biomarkers. Where possible 14 sediment samples were collected at random from each of 17 Shetland and 9 Orkney sites. The wide range of total PAH concentrations in sediments (i.e., < LOD up to 22619 ng g(-1) dry weight) was indicative of a variety of anthropogenic activities and different sediment types associated with the specific locations. Low PAH concentrations were determined in sandy sediments from areas of limited boat activity. The highest PAH concentrations were found in muddy sediment close to a boat repair yard. PAH concentration ratios were consistent with the main source of these compounds, in most areas, being pyrolysis. Geochemical biomarker (triterpane and sterane) profiles from the sediment were indicative, for some areas, of limited petrogenic input. Punds Voe was the only voe to show evidence of North Sea oil. PAH profiles were similar across sites within each island group, with any differences attributable to known local sources of PAHs. However, there was a clear difference in the PAH profiles of Shetland and Orkney sediments, with Orkney sediments having a higher proportion of the lighter alkylated PAHs.

Analysis of sediments from Shetland Island voes for polycyclic aromatic hydrocarbons, steranes and triterpanes

A few days after the grounding of the oil tanker Braer on 5 January 1993, an Exclusion Zone was designated by Order under the Food and Environment Protection Act 1985, prohibiting the harvesting of farmed or wild shellfish within the Zone to prevent contaminated products reaching the market place. The order was progressively lifted for species that were found to be free of petrogenic taint and for which the polycyclic aromatic hydrocarbon (PAH) levels were within the range for reference samples. This Order, however, still remains in place for mussels (Mytilus edulis) as the PAH levels are higher than in reference mussels. To investigate the possible source of PAHs found in these mussels, sediments were collected from three reference and three Zone sites and their hydrocarbon compositions studied using the n-alkane composition and concentration, PAH composition and concentration and the sterane and triterpane composition. The reference site at Olna Firth was found to have the highest levels of 2-6-ring parent and branched PAHs, the highest concentration in one of the pooled sediments being 4,530 ng g(-1) dry weight. Values in the other two reference sites (Vaila Sound and Mangaster Voe) ranged from 248.7 to 902.2 ng g(-1) dry weight. PAH concentrations at the Zone sites (Sandsound Voe, Stromness Voe and Punds Voe) ranged from 641.0 to 2,766 ng g(-1) dry weight. The PAH data were normalised to the percentage of organic carbon and log-transformed prior to being analysed using principal component analysis. The mean total PAH concentrations for Zone sites were found not to be significantly different from the reference sites. The PAH concentration ratios were consistent with the main source of PAHs being pyrolysis. However, there was a petrogenic contribution, suggested by the presence of alkylated PAHs, with Punds Voe having the largest petrogenic hydrocarbon content. This was supported by the triterpane profiles and the presence of a UCM in the aliphatic chromatograms from Punds Voe sediments.

Investigating the significance of dissolved organic contaminants in aquatic environments: Coupling passive sampling with in vitro bioassays

We investigated the feasibility of coupling passive sampling and in vitro bioassay techniques for both chemical and ecotoxicological assessment of complex mixtures of organic contaminants in water. Silicone rubber passive sampling devices (SR-PSDs) were deployed for 8-9 weeks in four streams and an estuary of an agricultural catchment in North East (NE) Scotland. Extracts from the SR-PSDs were analysed for freely dissolved hydrophobic organic contaminants (HOCs) and screened for wide range of pesticides. The total concentrations of dissolved PAHs (∑PAH(40), parent and branched) in the water column of the catchment varied from 38 to 69 ng L(-1), whilst PCBs (∑PCB(32)) ranged 0.02-0.06 ng L(-1). A number and level of pesticides and acid/urea herbicides of varying hydrophobicity (logK(OW)s ~2.25 to ~5.31) were also detected in the SR extracts, indicating their occurrence in the catchment. The acute toxicity and EROD induction potentials of SR extracts from the study sites were evaluated with rainbow trout liver (Oncorhynchus mykiss; RTL-W1) cell line. Acute cytotoxicity was not observed in cells following 48 h exposure to the SR extracts using neutral red uptake assay as endpoint. But, on a sublethal level, for every site, statistically significant EROD activity was observed to some degree following 72 h exposure to extracts, indicating the presence of compounds with dioxin-like effect that are bioavailable to aquatic organisms in the water bodies of the catchment. Importantly, only a small fraction of the EROD induction could be attributed to the PAHs and PCBs that were determined. This preliminary study demonstrates that the coupling of silicone rubber passive sampling techniques with in vitro bioassays is feasible and offers a cost effective early warning signal on water quality deterioration.

An assessment of persistent organic pollutants in Scottish coastal and offshore marine environments.

Concentrations of persistent organic pollutants (POPs), including polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) were determined in sediment and biota (fish liver) from around Scotland. The concentrations were investigated using assessment criteria developed by OSPAR and ICES. Organic contaminant concentrations, PAHs, PCBs and PBDEs in sediment, and PCBs and PBDEs in fish liver, were significantly higher in the Clyde compared to all other sea areas. This is mainly due to historic industrial inputs. Highest PCB and PAH concentrations were found in the strata furthest up the Clyde estuary, with concentrations of POPs in these strata being at levels such that there is an unacceptable risk of chronic effects occurring in marine species. Furthermore, for PAHs in Clyde sediment there was a significant negative gradient going from north to south towards the open sea. PAH and PCB concentrations in sediment and biota in all other Scottish sea areas (except for PCBs in sediment from East Scotland) were unlikely to give rise to pollution effects, being below relevant assessment criteria. Although no assessment criteria are available for PBDEs, the concentrations observed in Scottish sediments were low with all congeners below the limit of detection (LoD; 0.03 µg kg(-1) dry weight) in 140 out of a total of 307 samples analysed. Where PBDEs were detected, the dominant congeners were BDE47 and BDE99. PBDEs were detected in fish livers, although concentrations were less than 150 µg kg(-1) lipid weight in all sea areas except the Clyde where concentrations ranged between 8.9 and 2202 µg kg(-1) lipid weight. Few trends were detected in contaminant concentrations in biota or sediment at any Scottish site with more than five years data. Downward trends were detected in PAHs in sediment from the Clyde, Irish Sea and Minches and Malin Sea and PCBs in fish liver from the Moray Firth. Rules were developed for the aggregation of the contaminant data across a sea area. An overall assessment for each sea area was then assigned, looking at the frequency of sites or strata within each sea area that were above or below the relevant assessment criteria. Overall the status of the various sea areas, with respect to the assessed POPs, can be considered to be acceptable in that they were below concentrations likely to result in chronic effects for all sea areas except the Clyde.

Uptake and effects of the cypermethrin-containing sea lice treatment Excis in the marine mussel, Mytilus edulis.

Wild and farmed mussels, Mytilus edulis, coexist with salmon farms in Scottish sea lochs. A synthetic pyrethroid, cypermethrin, is licensed for use on fish farms to control sea lice infestations as a formulation called Excis. In this study, uptake of cypermethrin from Excis exposure is investigated through the use of gas chromatography with mass-spectrometry. The effects of Excis on mussels are also examined by measuring the neutral red retention time of lysosomes, aerial survival and shell closure. The isomeric ratios of cis:trans cypermethrin measured in mussels are around 80:20; a marked increase from 40:60 to which the mussels were exposed. This is most likely due to preferential metabolism of trans-isomers, as the same response is seen in vertebrates. There is a pronounced behavioural effect of shell closure, where mussels exposed to 1000 microg/l cypermethrin shut their shells within an hour of exposure. Arguments are presented for this effect being either a voluntary response on recognition of cypermethrin, or an effect arising from an involuntary action of cypermethrin on the adductor muscle. Even at 1000 microg/l cypermethrin, neutral red retention time and aerial survival are not affected. The data suggest that the responses of mussels shown here are unlikely to occur in the field, even at the concentrations of cypermethrin used in fish cages, for the treatment of sea lice.

Molecular responses of European flounder (Platichthys flesus) chronically exposed to contaminated estuarine sediments

Molecular responses to acute toxicant exposure can be effective biomarkers, however responses to chronic exposure are less well characterised. The aim of this study was to determine chronic molecular responses to environmental mixtures in a controlled laboratory setting, free from the additional variability encountered with environmental sampling of wild organisms. Flounder fish were exposed in mesocosms for seven months to a contaminated estuarine sediment made by mixing material from the Forth (high organics) and Tyne (high metals and tributyltin) estuaries (FT) or a reference sediment from the Ythan estuary (Y). Chemical analyses demonstrated that FT sediment contained significantly higher concentrations of key environmental pollutants (including polycyclic aromatic hydrocarbons (PAHs), chlorinated biphenyls and heavy metals) than Y sediment, but that chronically exposed flounder showed a lack of differential accumulation of contaminants, including heavy metals. Biliary 1-hydroxypyrene concentration and erythrocyte DNA damage increased in FT-exposed fish. Transcriptomic and (1)H NMR metabolomic analyses of liver tissues detected small but statistically significant alterations between fish exposed to different sediments. These highlighted perturbance of immune response and apoptotic pathways, but there was a lack of response from traditional biomarker genes. Gene-chemical association annotation enrichment analyses suggested that polycyclic aromatic hydrocarbons were a major class of toxicants affecting the molecular responses of the exposed fish. This demonstrated that molecular responses of sentinel organisms can be detected after chronic mixed toxicant exposure and that these can be informative of key components of the mixture.