Evidence of polycyclic aromatic hydrocarbon (PAH) exposure in fish from the Antarctic peninsula

PAH exposure was assessed in fish from Arthur Harbor, Antarctica and sites remote from human activities by measuring the concentrations of polycyclic aromatic hydrocarbon (PAH) metabolites in bile and PAH in tissues and stomach contents. The highest concentrations of biliary metabolites and tissue PAH were present in fish captured near an Argentine supply ship (Bahia Paraiso) that ran aground in Arthur Harbor in 1989. The presence of biliary PAH metabolites in fish collected near Palmer Station, a small US scientific station in Arthur Harbor, suggests low level exposure to contaminants. However, PAH tissue concentrations in fish from Palmer Station were similar to those from remote sites. Both tissue PAH and metabolite concentrations were low in fish captured at sites remote from human activities. The confirmation of PAH metabolites in the bile of fish from Arthur Harbor is direct evidence that the presence of humans has resulted in Antarctic fish being exposed to petroleum-derived PAH.     

Assessment of PAH Pollution in Coastal Areas from the NW Mediterranean through the Analysis of Fish Bile

Polycyclic Aromatic Hydrocarbon (PAH) pollution in coastal areas from the NW Mediterranean has been assessed by measuring bile PAH metabolites in two fish species, the red mullet – Mullus barbatus-and the sea comber – Serranus cabrilla – Bile crude samples were directly analysed by LC-fluorescence at the excitation/emission wavelengths of benzo[a]pyrene (380/430 nm). Subsequently, bile samples were hydrolysed and analysed by (a) LC-fluorescence at the excitation/emission wavelength pairs of naphthol (290/335 nm) and pyrenol (345/395 nm), and (b) gas chromatography–mass spectrometry (GC–MS) for the determination of individual PAHs. Qualitative and quantitative differences were recorded among sampling sites. Both species allowed us to detect different pollution gradients, the highest levels of FACs were observed in organisms caught in the vicinity of big industrial cities, whereas the lowest levels were recorded in organisms from Corsica and Sardinia. The obtained results highlight the usefulness of the LC-fluorescence technique as an initial screening method to assess PAH exposure in coastal fish.     

Assessing PAH exposure in feral finfish from the Northwest Atlantic

Polycyclic aromatic hydrocarbon (PAH) concentrations were examined in small finfish (<30 cm) represented by capelin, sand lance, American plaice, yellowtail flounder and herring collected opportunistically in various NAFO divisions. Analyses were performed on whole fish and in a portion of the samples; concentrations in internal organs were compared to the rest of the carcass. The effect of pool size, size differences within and between species, lipid content and location were examined to interpret PAH concentrations. Measurements were carried out before the development of the Hibernia oil fields and represent baseline levels for future comparison. Limits in assessing future risk that could be due to discharges of produced water or accidental oil spills are also discussed. Increasing knowledge on the bioaccumulation of PAH, on the production of bile metabolites, the formation of DNA-adducts and of the potential toxic effects associated with PAH will lead to better ecosystem management and protection for future generations.     

Biomarker responses in flounder (Platichthys flesus) and mussel (Mytilus edulis) in the Klaipeda-Būtinge area (Baltic Sea)

During the EU project BEEP a battery of biomarkers was applied in flounder (Platichthys flesus) and the blue mussel (Mytilus edulis) collected at three locations off the Lithuanian coast (Baltic Sea) in June and September 2001 and 2002. The elevated biomarker responses in specimens sampled in September 2001 were apparently related to the extensive dredging activities in the Klaipeda port area and subsequent dumping of contaminated sediments. High concentrations of organic pollutants (organochlorines and PBDEs) were also measured in the tissues of both indicator species. In addition, response levels of genotoxicity, cytotoxicity, immunotoxicity as well as concentrations of PAH metabolites in the bile of flounder showed elevations in 2002 after an oil spill in the Būtinge oil terminal in November 2001. In flounder, biomarker measurements 10 months after the spill indicated recovery processes but in mussels a high level of genotoxicity could still be observed 22 months later. The present study illustrates the usefulness of the multi-biomarker approach in the detection of biological effects of pollution in this region of the Baltic Sea.     

Biliary fluorescent aromatic compounds (FACs) measured by fixed wavelength fluorescence (FF) in several marine fish species from the NW Mediterranean

The fixed wavelength fluoresce (FF) method was used to estimate the levels of fluorescent aromatic compounds (FACs) in the bile of fourteen fish species of commercial and/or ecological interest. Sampling was carried out in the NW Mediterranean at depths ranging from 50 to 1000 m during four seasonal cruises. During the summer sampling period, some species were also collected from another site (Vilanova fishing grounds) for comparison. Baseline levels of the polycyclic aromatic hydrocarbons (PAHs) naphthalene, phenanthrene, pyrene and benzo[a]pyrene were measured. Some seasonality was observed, with reduced FF levels in summer and no differences among sites, consistent with sediment PAH levels. We discuss our results in relation to fish phylogeny, season, depth, diet, trophic level and swimming capacity. Overall FF levels indicated differences among species; the suprabenthic feeders from shallow and deep communities, and Mullus barbatus in particular, displayed elevated FF values and are potential candidates for additional monitoring studies.     

Non-destructive assessment of polycyclic aromatic hydrocarbon (PAH) exposure by fluorimetric analysis of crab urine

The detection of urinary polycyclic aromatic hydrocarbon (PAH) metabolites by fluorescence spectrophotometry is particularly effective as a practical means to assess PAH exposure in decapod crabs. However, the practical application of this technique has thus far only been tested for the European shore crab (Carcinus maenas) and only a few field studies have been conducted in heavily polluted areas. The present study evaluated the adaptability of this method as a rapid, cost-effective and non-destructive biomonitoring tool for the New Zealand crab species, Macrophthalmus hirtipes (stalk-eyed mud crab). A field gradient could be detected among the sites and different input sources of PAH contamination could be discerned through the differentiation of pyrogenic and petrogenic PAH signatures. The present study shows that the fluorescence screening method is sensitive to relatively low levels of PAH contamination and more broadly applicable to smaller crab species than C. maenas, for which the technique was developed.     

Mugil curema as a PAH bioavailability monitor for Atlantic west sub-tropical estuaries

This work aimed to evaluate the use of Mugil curema as a biomonitor for polycyclic aromatic hydrocarbon (PAH) bioavailability. Fish biliary metabolites from low-contaminated (Cananéia) and contaminated (Santos) Brazilian estuaries were analysed using a high-performance liquid chromatography apparatus coupled to fluorescence detectors connected in a series. In the Cananéia and Santos estuaries, total metabolite levels ranged from 0.85 to 34.4 μg g^? 1 of bile and from 4.06 to 528 μg g^? 1 of bile, respectively. Metabolite levels were not influenced by feeding status or sexual hormones (p < 0.05), thus suggesting that M. curema is a good biomonitor for PAH bioavailability. In estuarine regions, it is possible to sample both the male and female genders of this species in any reproductive period.     

Survey of estrogenic and androgenic disruption in Swedish coastal waters by the analysis of bile fluid from perch and biomarkers in the three-spined stickleback

The potential for endocrine disruption close to sewage treatment plant and pulp mill effluent discharge points along the Swedish Baltic Sea coast was explored using a dual survey strategy employing two stationary fish species. The levels of vitellogenin and spiggin as biomarkers of endocrine disruption were determined in juvenile three-spined sticklebacks (Gasterosteus aculeatus L.) together with the sex ratios and the presence of intersex. As an indication of exposure, estrogenic and androgenic substances were analysed by GC–MS in bile from perch (Perca fluviatilis L.). Spiggin and vitellogenin levels in juvenile three-spined sticklebacks were generally low, and, for most sampling sites no deviation in gonad type ratios were observed. No remarkable levels of natural or synthetic estrogens or androgens were observed in bile fluid from perch, while bisphenol A and 4-nonylphenol were detected in perch from both reference sites and exposed sites. Taken together, the results did not indicate estrogenic or androgenic disruption in the investigated waters.     

Indices for the assessment of environmental pollution of the Baltic Sea coasts: integrated assessment of a multi-biomarker approach

Two mathematical methods to assess the "health status" of flounder (Platichthys flesus), eelpout (Zoarces viviparus) and blue mussel (Mytilus spp.) populations of the Baltic Sea were applied on selected biomarker data collected during the EU project "BEEP" (Biological Effects of Environmental Pollution on Marine Coastal Ecosystems). The Bioeffect Assessment Index (BAI) and the Integrated Biomarker Index (IBR) combine different biomarkers to single values, which can be used to describe the toxically-induced stress level of populations in different areas. Both indices determined here produced essentially similar results, which in most cases agreed with the known contamination levels in the different study areas. Advantages and limitations of index applications and interpretations are critically discussed. The use of indices provides comprehensive information about biological effects of pollution in marine organisms and may therefore serve as a useful tool for environmental management by ranking the pollution status of marine coastal areas.     

Biomarkers of contaminant exposure: results of a field study with flounder (Platichthys flesus) from the southern Baltic Sea

The results of the present study are based on enzyme biomarker measurements in flounder (Platichthys flesus), a flatfish species that is widely distributed in Baltic coastal waters. The fish were collected from known pollution gradients and from reference areas regarded as relatively free of anthropogenic input. Muscular cholinesterases (ChEs), hepatic glutathione S-transferase (GST), and hepatic ethoxyresorufin-O-deethylase (EROD) activities were measured in each sampled specimen of flounder. Generalized linear models (GLM) were used to analyze the dependence of the enzyme activity on sampling year and area as well as on the biological parameters of the fish. Statistically significant differences in the activities of the measured biomarkers were observed between reference and contaminated sites. ChEs and GST activities differed with gender. The results of this study suggest that the location and year of sampling have a significant impact on the activity of all the measured biomarkers.     

Tributyltin in blood of marine fish collected from a coastal area of northern Kyushu, Japan

We investigated levels of the pollutant tributyltin (TBT) in blood of pufferfishes (six species), Japanese sea perch, red sea bream, Japanese common goby, Japanese flounder, rockfish, conger eel, and sea mullet collected off the coast of northern Kyushu, Japan. We found considerable levels of TBT (1.4-190 ng/mL) accumulated in the blood of these fish. Blood TBT concentrations were 1.3-22.5 times liver concentrations and 4.9-78 times muscle concentrations, except in conger eel and mullet. We detected TBT (16-111 ng/mL-blood) in the plasma of the fine-patterned puffer (Takifugupoecilonotus) year-round, without any apparent seasonal trend. These results suggest that fish inhabiting coastal areas of Kyushu, Japan, continue to be contaminated with TBT.     

Bioaccumulation and biomarker responses of trace metals and micro-organic pollutants in mussels and fish from the Lagoon of Venice,Italy

The aims of this study were to evaluate levels and effects of trace metals (Cd, Cr, Cu, Fe, Mn, Pb, Zn), PAHs and PCBs in mussels and fish native to the Lagoon of Venice. To this end, bioaccumulation and various biomarkers of generic stress and exposure (malondialdehyde, MDA and metallothioneins, MT; ethoxyresorufin O-deethylase activity, EROD; fluorescent bile metabolites and somatic indices in fish; lysosomal membrane stability, LMS; lipofuscins, LF; neutral lipids, NL and survival-in-air, SOS in mussels) were measured in organisms seasonally collected from two differently influenced areas.In mussels, metal bioaccumulation levels at the two sites were low and fluctuated similarly, exhibiting decreasing levels in summer, like MDA and MT. Micro-organic pollutants and the probably related biological response of LMS did not show either significant site differences or seasonal trends. LF and NL revealed inconclusive patterns. The results of the survival-in-air test were quite erratic, showing that they were associated with the high variability of both natural and physiological parameters temperature, food, reproduction, and body reserve cycle.In fish, both inorganic and organic micropollutant levels were low, and differences were detected between sites only for PCBs. The pattern exhibited by micro-organic contaminants was clearly related to the reproductive cycle at both sites; a relationship with PCBs, EROD and MDA was established at the site where their levels were generally higher. Bile metabolites (3-OH benzo(a)pyrene, 1-OH pyrene) were associated with PAH contents.     

Significance of cytochrome P450 system responses and levels of bile fluorescent aromatic compounds in marine wildlife following oil spills

The relationships among cytochrome P450 induction in marine wildlife species, levels of fluorescent aromatic compounds (FAC) in their bile, the chemical composition of the inducing compounds, the significance of the exposure pathway, and any resulting injury, as a consequence of exposure to crude oil following a spill, are reviewed. Fish collected after oil spills often show increases in cytochrome P450 system activity, cytochrome P4501A (CYP1A) and bile fluorescent aromatic compounds (FAC), that are correlated with exposure to polycyclic aromatic hydrocarbons (PAH) in the oil. There is also some evidence for increases in bile FAC and induction of cytochrome P450 in marine birds and mammals after oil spills. However, when observed, increases in these exposure indicators are transitory and generally decrease to background levels within one year after the exposure. Laboratory studies have shown induction of cytochrome P450 systems occurs after exposure of fish to crude oil in water, sediment or food. Most of the PAH found in crude oil (dominantly 2- and 3-ring PAH) are not strong inducers of cytochrome P450. Exposure to the 4-ring chrysenes or the photooxidized products of the PAH may account for the cytochrome P450 responses in fish collected from oil-spill sites. The contribution of non-spill background PAH, particularly combustion-derived (pyrogenic) PAH, to bile FAC and cytochrome P450 system responses can be confounding and needs to be considered when evaluating oil spill effects. The ubiquity of pyrogenic PAH makes it important to fully characterize all sources of PAH, including PAH from natural resources, e.g. retene, in oil spill studies. In addition, such parameters as species, sex, age, ambient temperature and season need to be taken into account. While increases in fish bile FAC and cytochrome P450 system responses, can together, be sensitive general indicators of PAH exposure after an oil spill, there is little unequivocal evidence to suggest a linkage to higher order biological effects, e.g. toxicity, lesions, reproductive failure.     

Bioaccumulation and biomarker responses of trace metals and micro-organic pollutants in mussels and fish from the Lagoon of Venice, Italy

The aims of this study were to evaluate levels and effects of trace metals (Cd, Cr, Cu, Fe, Mn, Pb, Zn), PAHs and PCBs in mussels and fish native to the Lagoon of Venice. To this end, bioaccumulation and various biomarkers of generic stress and exposure (malondialdehyde, MDA and metallothioneins, MT; ethoxyresorufin O-deethylase activity, EROD; fluorescent bile metabolites and somatic indices in fish; lysosomal membrane stability, LMS; lipofuscins, LF; neutral lipids, NL and survival-in-air, SOS in mussels) were measured in organisms seasonally collected from two differently influenced areas. In mussels, metal bioaccumulation levels at the two sites were low and fluctuated similarly, exhibiting decreasing levels in summer, like MDA and MT. Micro-organic pollutants and the probably related biological response of LMS did not show either significant site differences or seasonal trends. LF and NL revealed inconclusive patterns. The results of the survival-in-air test were quite erratic, showing that they were associated with the high variability of both natural and physiological parameters temperature, food, reproduction, and body reserve cycle. In fish, both inorganic and organic micropollutant levels were low, and differences were detected between sites only for PCBs. The pattern exhibited by micro-organic contaminants was clearly related to the reproductive cycle at both sites; a relationship with PCBs, EROD and MDA was established at the site where their levels were generally higher. Bile metabolites (3-OH benzo(a)pyrene, 1-OH pyrene) were associated with PAH contents.     

Liver histopathology in Baltic flounder (Platichthys flesus) as indicator of biological effects of contaminants

Results are presented of a study on liver histopathology in Baltic flounder (Platichthys flesus) carried out in 2001 and 2002 in four coastal sampling areas of the Baltic Sea: Kv?d?fj?rden (Swedish east coast, reference area), Klaipeda-Butinge (Lithuanian coast), Gulf of Gdansk (Polish coast), and Wismar Bay (German coast) within the framework of the EU-funded BEEP project. Liver lesions were diagnosed and categorised using standardised methodologies and, for a spatial and temporal assessment of the prevalence and types of lesions detected, a scoring system was applied, involving the calculation of mean histopathology lesion scores. 83.0% of the 436 female flounder examined (size range: 20-43 cm total length, age range: 2-8 years) were affected by liver lesions, out of which 74.3% were assigned to the category of non-specific, 3.4% to the category of early toxicopathic non-neoplastic, 4.6% to the category of pre-neoplastic and 0.7% to the category of neoplastic lesions. Mean lesions scores were highest in the areas at the Lithuanian and Swedish coast and there is indication of an impact of the age structure of the flounder populations studied, the sampling season as well as of contaminant effects.     

Biomarker responses in pelagic and benthic fish over 1 year following the Hebei Spirit oil spill (Taean, Korea)

After the Hebei Spirit oil spill incident (7th December, 2007) in the west coast of Korea, contamination of biliary PAH metabolite and hepatic biomarkers in a pelagic and a benthic fish was monitored for 1 year. Concentrations of 16 PAHs and alkylated PAHs in fish muscle were highest (22.0 ng/g d.w. for 16 PAHs and 284 ng/g d.w. for alkylated PAHs) at 5 days after the spill and then decreased rapidly to background levels at 11 months after the spill. Fish from the oiled site had elevated biliary PAH metabolite concentrations immediately after the spill; these declined steadily in both species, but were still above reference site concentrations 2 months after the spill. Oiled-site fish showed hepatic CYP 1A induction whose trend closely followed those of biliary PAH metabolite concentrations, implying continuous exposure to PAHs. Brain acetylcholinesterase activity was not related to oil exposure.     

Developmental disorders in larvae of eelpout (Zoarces viviparus) from German and Swedish Baltic coastal waters

Pregnant eelpout were collected for several years at German and Swedish Baltic coastal stations with different impact of pollutants and the prevalence of fry abnormalities compared between sites. Mortality of apparently normal developed larvae was observed with high variability in both German and Swedish sites. Malformation of larvae was a prominent disorder at all German stations with a prevalence ranging from about 50% up to a maximum of 90% in almost all analysed samples. At the Swedish stations malformation rates were significantly lower (range: 0-6%). Growth retardation of fry was clearly more prominent in eelpout from German coastal waters as well. Taken together the field data suggests that impaired larval development was associated with the state of environmental pollution at the sampling locations.     

Toxic contaminants and their biological effects in coastal waters of Xiamen,China. I. Organic pollutants in mussel and fish tissues

Organochlorines and organophosphates were measured in four fish species (Lateolabrax japonicus, Pagrasomus major, Miichthys miiuy and Epinephalus awoara) and the mussel Perna viridis collected from aquaculture cages in coastal waters of Xiamen, China. Polycyclic aromatic hydrocarbon (PAH) metabolites were also measured in fish bile. Sites in Xiamen coastal waters (the harbour, Maluan and Tongan) are compared with a "cleaner" reference site at Dongshan Island. DDT was found in livers of all fish sampled (May 1998, 1999 and December 1999) ranging from 0.15 to 2.2 microg/g WW, but levels in muscle tissue (<0.5 ng/g to 0.22 microg/g WW) were at least an order of magnitude lower in the harbour, and were not detectable (<0.5 ng/g) in Tongan samples. All other pesticides examined were not detected in fish or mussels, except for dieldrin in one liver sample (0.07 microg/g WW). Food of the caged fish, small fish and dried pellets, had very low (0.015-0.027 microg/g WW) or non-detectable levels of DDT, indicating significant bioaccumulation of contaminants in caged fish and/or other possible sources of uptake (water and sediments). DDT levels in fish varied with species. DDT contamination in fish livers and whole mussels varied significantly with location. Highest concentrations in fish occurred in the harbour, and less consistently so in Tongan. Dongshan and Maluan had comparatively low levels of DDT. Relatively low residues of DDT in both fish and mussels at Dongshan likely reflect lower levels of pesticide input at this reference site. However, the low levels of DDT contamination in fish tissues from Maluan were unexpected in view of the close proximity of the harbour and possibly reflect the removal of particulate-bound contaminants by the intensive bivalve culture there. Consistently high proportions of DDT in the form of the parent compound (p,p'-DDT: fish 30-45%; mussels 40-65%) from all sites suggest recent releases of this chemical to the environment. Levels of two main groups of PAH metabolites (naphthalene and phenanthrene) in fish bile suggest a different environmental distribution pattern than that of pesticides, in that Dongshan fish appear to be no less exposed to hydrocarbons than those from Xiamen coastal waters.     

Monitoring polycyclic aromatic hydrocarbons in the Northeast Aegean Sea using Posidonia oceanica seagrass and synthetic passive samplers

The concentrations of 22 polyaromatic hydrocarbons (PAH) in Posidonia oceanica seagrass, sediments, and seawater from the Alexandroupolis Gulf in the Aegean Sea, were investigated from 2007 to 2011. Temporal trends of total PAH contents in P. oceanica and sediments were similar. PAH levels in seawater, sediments, and seagrasses generally decreased with increasing distance from Alexandroupolis Port. Leaves and sheaths of P. oceanica had higher PAH levels than roots and rhizomes. P. oceanica accumulates PAHs and has good potential as a bioindicator of spatiotemporal pollution trends. PAH concentrations were also examined using in situ passive seawater sampling and were compared to results of passive sampling in the laboratory using local sediments and seawater. Levels of high molecular weight PAHs assessed using passive samplers confirmed the decreasing gradient of pollution away from Alexandroupolis Port. Passive sampling also proved useful for investigating sources of PAHs in P. oceanica meadows.     

Biomarkers of exposure and effect in flounder (Platichthys flesus) exposed to sediments of the Adriatic sea.

The modifications of several biomarkers were investigated in flounder (Platichthys flesus) when exposed in the laboratory to sediment samples collected from the Northern Adriatic Sea. Besides clean sand used as a control substrate, fish were exposed to sediments sampled offshore the delta of the Po River, the harbour of Trieste, and from the industrial harbour of Venice (Porto Marghera). After six days of exposure, the enzyme activities ethoxyresorufin-O-deethylase (EROD), UDP glucuronyltransferase (UDPGT), glutathione S-transferase (GST), glutathione reductase (GR) and glutathione peroxidase (GPx), were assayed in fish liver. In addition, the contents of reduced glutathione (GSH), nonprotein thiols (SH), total sugars and extractable lipids were also determined in hepatic tissue, as well as the number of micronucleated hepatocytes and biliary concentrations of fluorescent aromatic compounds (FAC). Despite some variability within treatment groups, differences among exposed organisms were evident and consistent with known contaminant levels of sampled areas. Microsomal activities (EROD, UD-PGT) and FAC levels were the most sensitive exposure indicators. Variations in the other biomarkers showed only tendencies which although not statistically significant were generally consistent with the contamination pattern.