Polycyclic aromatic hydrocarbons in water, seston and copepods in a harbour area in the Western Mediterranean (Ligurian Sea)

Polycyclic aromatic hydrocarbons (PAHs) are widespread carcinogenic, mutagenic and teratogenic pollutants that can significantly affect ecosystems and exert an influence on marine planktonic and benthic communities as well as on higher organisms such as fish and mammals, including humans. In this paper the concentrations of six PAHs in water, seston and copepods were examined in a harbour zone of the Genova Gulf (Ligurian Sea) from November 1997 to June 1998. The importance of freshwater inputs coming from a polluted stream and the influence of seasonal and atmospheric factors were also considered. Highest PAH values were recorded during spring. Fluoranthene, Benzo[b]fluoranthene and Benzo[k]fluoranthene were the most abundant PAHs either in water, seston or copepods. The concentration of total PAHs in seawater was often lower than the detection limit: recorded values ranged between 0.001 and 0.06 ppb. In seston, total PAHs ranged between 0.033 and 6.26 ng·g^?1 and in copepods between 0.01 and 2.34 ng·g^?1. With this study, PAH contamination in the harbour zone of Genova Sestri‐Ponente and the importance of inputs of organic pollutants coming from land and freshwaters have been emphasized.     

Evidence for resistance to benzo[a]pyrene and 3,4,3'4'-tetrachlorobiphenyl in a chronically polluted Fundulus heteroclitus population

Halogenated aromatic hydrocarbons (HAHs) and polynuclear aromatic hydrocarbons (PAHs) are major environmental contaminants. Fish species that are chronically exposed to these compounds can develop resistance to their toxic effects. In all fish species studied to date, toxicant resistance has been accompanied by decreased inducibility of the xenobiotic metabolizing enzyme, cytochrome P450 1A (CYPIA). CYP1A induction is mediated through the Aryl Hydrocarbon Receptor (AHR). Although these compounds mediate their effects through this pathway, there have been resistant populations in which one chemical class cannot induce CYPIA expression (HAHs) while the other (PAHs) can. Resistance to PAHs was examined in a HAH-resistant population of Fundulus heteroclitus collected from a site contaminated with both compound classes (Newark Bay, NJ). Fish were injected intraperitoneally with the HAH 3,4,3',4'-tetrachlorobiphenyl (PCB77), benzo[a]pyrene (B[a]P, a PAH) or vehicle and sacrificed after 2 (B[a]P) or 5 days (PCB77, vehicle). We found no significant increase in CYP1A mRNA levels in resistant Newark Bay F. heteroclitus treated with either B[a]P or PCB77, while there was a 3.9 fold (PCB77) and 4.2 fold (B[a]P) increase in CYP1A mRNA in Flax fish relative to controls. AHR labeling studies revealed significantly (P < 0.05) lower levels of hepatic AHR in Newark fish (1,770 +/- 1,693.2 DPM) relative to Flax fish (6,082.5 +/- 1,709.9 DPM). Overall, these data suggest Newark F. heteroclitus are resistant to both PAHs and HAHs at the level of CYP1A mRNA, which might be mediated, in part, though lower expression of AHR. We are currently studying the promoter sequence to determine its role in chemical resistance.     

固相萃取-高效液相色谱-荧光检测法测定海水中的多环芳烃

建立了用固相萃取-高效液相色谱-荧光检测法测定海水中多环芳烃的方法,优化了色谱条件和萃取条件。除苊不能用荧光检测器检出外,其余15种多环芳烃的空白加标回收率在64.5%(苯并[g,h,i],茚并[1,2,3-cd]芘)～88.7%(苯并[a]蒽)之间,相对标准偏差(n=5)为4.9%(荧蒽,苯并[b]荧蒽)～11.1%(苯并[g,h,i],茚并[1,2,3-cd]芘),方法的检出限在0.72(蒽)～14.10 ng/L(荧蒽)之间,基本上达到了痕量分析的要求。利用该方法测得青岛湾表层海水中多环芳烃的浓度在0.125(苯并[k]荧蒽)～25.996 ng/L(萘)之间,但苯并[a]芘未检出。     

Photoinduced toxicity single and binary mixtures of four polycyclic aromatic hydrocarbons to the marine diatom Skeletonema costatum

Diatom Skeletonema costatum Cleve is one of the main predominant phytoplankton species in the Changjiang Estuary in China. In order to provide some basic information for future assessment of the potential risk on phytoplankton communities in this estuary caused by polycyclic aromatic hydrocarbons(PAHs), this alga was selected as a representative to investigate the photoinduced toxicity of PAHs, in single and mixture. Four PAHs including three-ring phenanthrene and anthracene, four-ring fluoranthene and pyrene were tested in the laboratory. The single toxicity of each PAH on this microalga was compared with and without the simulated solar UV radiation. The results showed that this microalga was sensitive to PAH's photoinduced toxicity. Ratios of the 72 h median effect concentration obtained for fluorescent and UV light tests were about 8.4 for phenanthrene, 13.0 for anthracene, 6.5 for fluoranthene, and 5.7 for pyrene, indicating that UV light enhanced the PAH toxicity to this alga significantly. Under the fluorescent radiation (lacking UV), the dose-response curves based on chemical concentrations revealed that the order of toxic strength was fluoranthene greater than pyrene greater than anthracene greater than phenanthrene; while under the UV radiation (476 μW/cm2 for UVA, 6.5 μW/cm2 for UVB) it became fluoranthene approximately equaling anthracene greater than pyrene greater than phenanthrene, indicating that the UV light also changed its relative toxicity to this alga. The photoinduced toxicity of PAHs to the marine diatom S. costatum might be a synergistic effect of photosensitization reactions (e.g., generation of single-state oxygen) and photomodification (photooxidation and/or photolysis).The combined effects of six binary mixtures on the marine diatom S. costatum were investigated using the additive-index method. Four binary-mixtures (phenanthrene plus anthracene; phenanthrene plus pyrene; anthracene plus fluoranthene; anthracene plus pyrene) were found to be synergistic joint action mode, while two binary-mixtures (phenanthrene plus fluoranthene; fluoranthene plus pyrene) displayed antagonist, revealing a complex pattern of possible interactions. The combined action mode of PAHs might be related to various factors such as the mixture compounds, mixture ratios and test conditions, etc.     

PAH-biodegradation potential of indigenous microorganisms: evidence from the respiratory activity of surface sediments in the Quanzhou Bay in China

Seven stations were established in the Quanzhou Bay (24.73°-24.96°N, 118.50°-118.70°E) in China on three cruises to determine the concentrations of polycyclic aromatic hydrocarbons (PAHs) and the numbers of PAH-degrading bacteria in surface sediments. Assessing the biodegradation poten- tial of indigenous microorganisms by measuring the respiratory intensity with the addition of PAHs in sediment samples was also one of the aims of this study. The results show that the total PAH concentrations of the sedimen...     

南大西洋沉积物中多环芳香化合物的组成特征研究

10 samples of sediments obtained from the South Mid-Atlantic Ridge were measured for the abundances and distributions of polycyclic aromatic compounds(PAHs). The total concentrations of PAHs(∑PAHs) ranged from 2.768 to 9.826 μg/g dry sediment. The ∑PAHs was higher in sample 22V-TVG10 and sample 26V-TVG05 which were close to hydrothermal fields, with the lowest value in sample 22V-TVG14 which was farthest from hydrothermal fields, suggesting a probable hydrothermal origin of ∑PAHs of samples. Approximately nine kinds of PAHs were identified, and low molecular mass tricyclic and tetracyclic aromatic compounds were predominant in the samples. The concentrations of fluoranthene which were typical as hydrothermal alteration compounds were the highest among PAHs with dry weight between 0.913–3.157 μg/g. The phenanthrene homologue was most abundant in the samples, and the ratios between parent phenanthrene and methylphenanthrene which probably reflected the degree of hydrothermal alteration ranged from 0.097 to 1.602. The sample 22V-TVG10 possessing a maximum ratio value showed the intense influence of the hydrothermal alteration on this sample, which might further imply that PAHs in sediments were mainly derived from the hydrothermal alteration.     

Induction of cytochrome P4501A and biliary PAH metabolites in European eel Anguilla anguilla: Seasonal, dose- and time-response variability in field and laboratory conditions

Induction of cytochrome P4501A in fish is a well-known indicator of exposure to polycyclic aromatic hydrocarbons (PAHs) and determination of PAH metabolites in bile by fixed wavelength fluorescence (FF) or synchronous fluorescence spectroscopy (SFS), has become an useful method in monitoring programs. In this work the relationship between cytochrome P4501A (EROD activity) and levels of biliary PAH metabolites was measured in the European eel Anguilla anguilla, in both field and laboratory conditions: organisms were sampled on a seasonal basis from the Orbetello lagoon (Tuscany) to characterize the natural variability of these biological parameters, while in laboratory eels were intraperitoneally injected with benzo[a]pyrene to investigate temporal and dose-dependent induction patterns. Results showed that induction of cytochrome P450 and accumulation of PAHs metabolites in bile are not necessarily correlated either in field, or in laboratory investigations; different seasonal changes were measured in natural conditions and slight variations in dose and time response patterns were also obtained in laboratory exposures.     

Induction of cytochrome P4501A and biliary PAH metabolites in European eel Anguilla anguilla: Seasonal, dose- and time-response variability in field and laboratory conditions

Induction of cytochrome P4501A in fish is a well-known indicator of exposure to polycyclic aromatic hydrocarbons (PAHs) and determination of PAH metabolites in bile by fixed wavelength fluorescence (FF) or synchronous fluorescence spectroscopy (SFS), has become an useful method in monitoring programs. In this work the relationship between cytochrome P4501A (EROD activity) and levels of biliary PAH metabolites was measured in the European eel Anguilla anguilla, in both field and laboratory conditions: organisms were sampled on a seasonal basis from the Orbetello lagoon (Tuscany) to characterize the natural variability of these biological parameters, while in laboratory eels were intraperitoneally injected with benzo[a]pyrene to investigate temporal and dose-dependent induction patterns. Results showed that induction of cytochrome P450 and accumulation of PAHs metabolites in bile are not necessarily correlated either in field, or in laboratory investigations; different seasonal changes were measured in natural conditions and slight variations in dose and time response patterns were also obtained in laboratory exposures.     

Differential gene expression in mummichogs (Fundulus heteroclitus) following treatment with pyrene: comparison to a creosote contaminated site

Mummichogs, Fundulus heteroclitus, an estuarine fish with a relatively small home range found along the eastern coast of the United States are well-suited to monitoring contaminant effects, including those of polycyclic aromatic hydrocarbons (PAHs). One of the common PAHs in estuaries is pyrene. We report here on efforts to develop multiple biomarkers of pyrene exposure in this species. Adult male mummichogs were exposed in the laboratory to the weak aryl hydrocarbon receptor (AhR) agonist pyrene at 0, 30, or 50 microg/L in 7-day static renewal exposures. The RNA was extracted from livers and alterations in mRNA expression were assessed by subtractive hybridization and differential display in order to produce multiple biomarkers of pyrene exposure. Genes demonstrating differential expression were confirmed by quantitative-PCR (Q-PCR) and include cytochrome P-450 1A (CYP1A), a putative hepatocyte growth factor activator, a X-ray inducible retrotransposon, and several expressed sequenced tags (ESTs). Some of these genes represent new biomarkers of pyrene exposure and potential biomarkers of PAH exposure. Therefore, similar changes were investigated at a Superfund site in Charleston, SC. Mummichogs from a creosote contaminated site and from a reference site (North Inlet National Estuarine Research Reserve near Georgetown, SC) were trapped, RNA extracted from the livers, and Q-PCR performed. Many of the genes differentially expressed following pyrene exposure were not altered at the creosote contaminated site in comparison to the reference site. However, CYP1A and an EST were induced. CYP1A induction at Diesel Creek indicates that this population of fish does not demonstrate refractory CYP1A phenotypes observed at several sites with high levels of AhR agonists. Ultimately, we anticipate that the use of multiple biomarkers of PAH exposure will provide useful information on the potential effects of toxicants.     

夏季珠江口水体中多环芳烃的分布、组成及来源

利用1999年7月对珠江口海域的调查资料,对该区表层海水中优控多环芳烃的分布、组成及来源进行了分析和讨论,结果表明:(1)夏季珠江口海域表层海水中14种溶解态多环芳烃[苊、芴、菲、蒽、荧蒽、芘、苯并(a)蒽、艹屈、苯并(b)荧蒽、苯并(k)荧蒽、苯并(a)芘、二苯并(a,h)蒽、苯并(g,h,i)艹北、茚并(1,2,3-cd)艹比]的质量浓度为63.8~171.7 ng/L,且沿着冲淡水流向呈降低趋势;(2)颗粒态中15种多环芳烃[萘、苊、芴、菲、蒽、荧蒽、芘、苯并(a)蒽、艹屈、苯并(b)荧蒽、苯并(k)荧蒽、苯并(a)芘、二苯并(a,h)蒽、苯并(g,h,i)艹北、茚并(1,2,3-cd)艹比]的质量浓度为60.7~186.7 ng/L,其分布与水体载沙量及悬浮颗粒物的性质、粒径有关,具有从河口内向外海降低的分布特征;(3)多环芳烃组成和特征参数比值的分析表明,珠江口海域高温裂解来源的多环芳烃在伶仃洋海区输入最多,且主要为人类活动中煤燃烧排放的,而在香港岛周围海区的输入则相对较少,且主要为油燃烧排放的;(4)与法国塞纳河及长江口等河口相比,珠江三角洲海域水体中存在高菲含量排放源。     

多环芳烃化合物对栉孔扇贝组织芳烃羟化酶活力的影响

研究了苯并(a)芘(B[a]P)、苯并(k)荧蒽(B[k]F)以及两者的混合物对栉孔扇贝(Chlamys farreri)鳃丝和消化盲囊芳烃羟化酶(AHH)活力的影响。结果表明,多环芳烃化合物(PAHs)对栉孔扇贝鳃丝和消化盲囊AHH活力具有显著的诱导作用(P<0.05),并且呈现了明显的时间、剂量效应特征。B[a]P和B[k]F混合物处理组鳃丝和消化盲囊的AHH活力在实验期间呈现先升高后稳定趋势,B[a]P处理组则表现峰值变化,然后呈下降趋势。消化盲囊各处理组AHH活力显著高于鳃丝。作者认为可以采用消化盲囊的AHH活力来反映周围环境PAHs的污染,并应以最大饱和浓度为标准。3种PAHs中B[a]P的毒性最强,PAHs对栉孔扇贝的毒性大小与PAHs的组成有关,不能以不同种类单一PAHs毒性的总和来代表PAHs混合物的毒性。     

海洋微生物对多环芳烃的降解

从海域沉积物中富集分离出以芘作为唯一碳源和能源的海洋微生物，以ST4富集培养的混合微生物作为研究对象；该海洋混合菌株能利用菲（Phe）、芘（Pyr)、荧蒽（Fla)等多种多环芳烃；在不同浓度的芘的降解中，当芘的浓度为50mg/dm^3时，其生长水平和降解速率最高；当芘的浓度为200mg/dm^3时，其生长受到抑制，芘几乎不能被降解。外加营养盐酵母浸出液和葡萄糖促进降解微生物的生长，提高降解速率。研究表明了海洋微生物在多环芳烃污染环境的生物修复应用前景。     

1-Hydroxypyrene as a biomarker of PAH exposure in the marine polychaete Nereis diversicolor

The possibility of using the pyrene metabolite 1-hydroxypyrene as a biomarker of polycyclic aromatic hydrocarbons (PAHs) exposure was investigated by exposure of the marine polychaete Nereis diversicolor to several PAHs in the laboratory. Animals were exposed to pyrene alone and to five different PAHs – phenanthrene, anthracene, pyrene, benzo[a]pyrene and benzo[k]flouranthene. After five days of exposure the concentrations of parent PAHs and 1-hydroxypyrene were identified using three different analytical methods, high-performance liquid chromatography with fluorescence detection (HPLC/F), synchronous fluorescence spectroscopy (SFS) and gas chromatography with mass spectrometric detection (GC/MS). The SFS measurements of 1-hydroxypyrene were validated by the more sensitive method HPLC/F. The positive correlation between total PAHs and 1-hydroxypyrene concentrations in the polychaete tissues observed in experiments, suggests the feasibility of 1-hydroxypyrene as a suitable biomarker for total PAH exposure assessment. Furthermore, the possibility of employment of the simple and rapid SFS method instead of HPLC/F for biomarker analysis has been confirmed by the positive and significant correlation between results achieved by these two analytical methods.     

Inhibition by polycyclic aromatic hydrocarbons of ATPase activities in Sebastiscus marmoratus larvae: relationship with the development of early life stages

Sebastiscus marmoratus larvae were exposed to waterborne polycyclic aromatic hydrocarbons (PAHs) containing 3-5 rings, benzo[a]pyrene (BaP), pyrene (Py) or phenanthrene (Phe), respectively at 0.01, 0.1 and 1 μg L?1. Cumulative mortality, frequency of dorsal curvature and rate of pericardial and yolk sac edema in larvae treated for 8 days were significantly increased in a dose-dependent manner. All three PAHs resulted in reduction of the lower jaw, and the extent of reduction increased with increasing concentrations of the PAHs. Na+/K+-ATPase and Ca2+-ATPase activity in larvae exposed to the three PAHs were all significantly inhibited in a dose-dependent manner. Analysis using the Pearson correlation coefficient indicated a significant correlation between the rate of the dorsal curvature and edema and the inhibition of Na+/K+-ATPase and Ca2+-ATPase activity, suggesting that the developmental defects caused by PAHs were related to their inhibition of Na+/K+-ATPase and Ca2+-ATPase activity.     

Pathological reactions and recovery of hepatopancreatic digestive cells from the marine snail Littorina littorea following exposure to a polycyclic aromatic hydrocarbon

The aim of this study was to investigate the cellular pathological responses of hepatopancreatic digestive cells from the periwinkle Littorina littorea exposed to the polycyclic aromatic hydrocarbon (PAH) fluoranthene and to ascertain whether any injurious effects were reversible within the experimental time scale. A secondary objective was to establish the relationship of the various reactions to animal health status, using lysosomal stability as an index of well-being. Exposure of snails to a concentration of 335 microgl(-1) (1.7 microM) fluoranthene (seawater renewed and spiked daily with fluoranthene) for 5 days resulted in a reduction in lysosomal stability (neutral red retention) and endocytosis; and an increase in smooth endoplasmic reticulum (ER) and 7-ethoxycoumarin-o-deethylase (ECOD; measured as cyano-ECOD) activity measured in isolated live digestive cells. Exposed snails treated with clean seawater for a further 8 days resulted in a return to control levels of lysosomal stability, ECOD and ER; endocytosis showed only a partial recovery. Multi-variate and uni-variate analysis showed that there were strong correlations between the various cellular biomarker responses. These findings are interpretable within the current framework of molluscan biomarker responses to PAHs. Principal component analysis was used to derive the first principal component for endocytosis, ER and ECOD reactions and these were plotted against lysosomal stability as a measure of cellular well-being. The resulting significant regression represents the mapping of the individual biomarkers within health status space for a gradient of fluoranthene toxicity. From this analysis, we concluded that endocytosis is an indicator of healthy snails while proliferation of ER and to a lesser extent induced ECOD are indicative of dysfunction and reduced health. Finally, the results indicate that stress induced by chronic exposure to a PAH is reversible.     

Stress protein expression in Ampelisca abdita (Amphipoda) exposed to sediments from San Francisco Bay

Stress proteins (heat shock proteins, hsps) form part of the cellular protein repair system, and are induced by a wide variety of Stressors. To determine their suitability as tools for assessing sublethal sediment toxicity, we measured levels of members of the stress protein families hsp60 and hsp70 in benthic estuarine amphipods (Ampelisca abdita) exposed to sediments from 23 different sampling sites in San Francisco Bay for 10 d. Concentrations of sediment-associated xenobiotics were determined. Per cent survival was recorded and surviving animals were analysed for stress proteins using western blotting techniques. An inverse correlation (r² = 0.44) was seen between amphipod survival and hsp64 levels, and hsp64 levels were positively correlated with concentrations of total polycyclic aromatic hydrocarbons (PAHs) (r² = 0.5). Principal component analysis revealed that amphipod mortality was linked to a combination of several PAHs (phenanthrene, fluoranthene, pyrene, benzo(a)pyrene) and di-n-butylphthalate at southern San Francisco Bay sites. At northern San Francisco Bay sites, negative correlations were found between hsp64 levels and organotin compounds (MBT, DBT, TBT), and between hsp71 levels and the PAHs, benzo (b,k)fluoranthene and benzo(G,H,I)perylene, suggesting an inhibitory effect of these compounds on stress protein expression.     

Polycyclic aromatic hydrocarbons in recent sediments from a subtropical estuary in Brazil

The scope of this work was to evaluate the levels of polycyclic aromatic hydrocarbons (PAHs) and their possible sources in two shallow sediment cores from an estuarine area located in the south region of Brazil, the Guaratuba Bay. The estuary is encompassed by an Environmental Protection Area (EPA) of 1306 km², which is still considered to be pristine, despite recent urban growth. To assess levels of 14 selected PAHs in the sediments, among those listed as “priority pollutants” by the US Environmental Protection Agency, the cores were divided in several sections, followed by extraction, clean-up, fractionation and subsequent analysis using HPLC coupled to a fluorescence detector. Among the PAHs detected, phenanthrene and fluoranthene showed the highest concentrations. PAHs with more than four rings were usually present in low concentrations or were undetectable. To assist in the identification of possible sources, the ratios An/(An + Phe) and Flt/(Flt + Py) (anthracene to anthracene plus phenanthrene and fluoranthene to fluoranthene plus pyrene) and a PCA model were also used. Total concentrations of PAHs ranged between 1.5 and 3130 ng/g (mean 495) dry weight for the core collected in the inner part of the estuary, and between 78.5 and 3270 ng/g (mean 899) dry weight for the other core, collected in the outer part. Compared to values found in other studies in coastal zones along South American eastern margin, this area can be considered to range from low polluted to moderately polluted. From the data, it was also possible to conclude that there is predominance of petroleum sources, and essentially close to the more urbanised areas. Combustion sources have only minor contribution and are episodic, when compared to the latter.     

Phototoxicity of pyrene and benzo[a]pyrene to embryo-larval stages of the Pacific oyster Crassostrea gigas

There is a growing body of evidence to suggest that certain polycyclic aromatic hydrocarbons (PAHs) pose a greater hazard to aquatic organisms than previously demonstrated, due to their potential to cause photo-induced toxicity when exposed to ultraviolet (UV) radiation. The consequences of photo-induced toxicity are reported here for embryo-larval stages of the pacific oyster Crassostrea gigas, following exposure to pyrene and benzo[a]pyrene. During laboratory investigations, significant increases in toxicity were observed in the presence of environmentally attainable levels of UV-radiation, compared with embryos exposed to PAH alone, at levels previously deemed to have little acute biological effect. The phototoxicity of pyrene and benzo[a]pyrene completely inhibited the development to the D-shell larval stage when embryos were simultaneously exposed to 5 microg l(-1) PAH and ultraviolet light (UVB = 6.3 +/- 0.1 microW/cm2 and UVA = 456.2 +/- 55 microW/cm2). A linear relationship was also demonstrated for benzo[a]pyrene phototoxicity with decreasing UV light intensity.     

Polycyclic aromatic hydrocarbons in surface sediments of the Zhelin Bay in the South China Sea

This study examined the concentrations of polycyclic aromatic hydrocarbons (PAHs) in surface sediments collected in July 2004 from eight stations in the Zhelin Bay, one of the most important bays for large-scale mariculture in Guangdong Province. Thirteen individual parent PAH compounds were identified using high performance liquid chromatography with UV detection. The overall average concentration of total PAHs was 477.0 ng/g, ranging from 146.1 to 928.8 ng/g. Low molecular mass PAHs with two to three rings (e.g., acenaphthene) were dominant in each sample. The PAH concentration varied among sampling stations, with the highest concentration observed at bay outlets and the lowest found at stations outside the bay. Ratios of low to high molecular mass PAHs and fluoranthene to pyrene were used to determine the origin of PAHs, and results indicated mainly petroleum-derived contamination. Compared with other bays and harbors around the world, the total concentrations of PAHs in surface sediments at the Zhelin Bay are moderate, but this does not exclude the possibility of potential impact on human consumers because some strong carcinogenic PAHs with high molecular mass were found at the station with a nearby caged-fish and oyster farm. Long-term monitoring of PAH contamination in the Zhelin Bay is recommended to reduce the potential toxicological effects on aquatic organisms and humans.     

Different levels of mussel (Mytilus edulis) DNA strand breaks following chronic field and acute laboratory exposure to polycyclic aromatic hydrocarbons

Levels of polycyclic aromatic hydrocarbons (PAHs) including benzo[a]pyrene (B[a]P) were at least seven-fold higher in mussels sampled from a polluted site (Loch Leven, in Scotland, UK) compared to a nearby clean reference site (Loch Etive) throughout the year 2000. Levels of DNA strand breaks (alkaline COMET assay) using both gill and digestive gland nuclei were similar at both sites despite the difference in contaminant load (total PAH). In contrast, mussels collected from a reference site (Port Quin, Cornwall, UK) had an increase in DNA strand breaks in digestive gland cells following laboratory exposure to B[a]P-dosed Isochrysis galbana. However, after 14 days high dose (20 ppb-exposed diet) animals had returned to levels similar to the controls. There was no evidence of increased necrosis or apoptosis after treatments. The results from these two studies suggest that an adaptive response may prevent ongoing DNA damage in mussels exposed to high levels of B[a]P and PAH contamination.