The occurrence of hydrocarbons and histopathological abnormalities in oysters for seven years following the wreck of the Amoco Cadiz in Brittany (France)

A study of the main oyster areas polluted by the Amoco Cadiz oil spill (Abers, Baie de Morlaix) was carried out between 1978 and 1985 by chemical analyses (mainly aromatic hydrocarbon determinations) and histopathological observations of lesions in the digestive tracts and gonads of oysters. The hydrocarbon content in oyster tissues was monitored and analyzed by UV spectrofluorimetry, and gas and liquid chromatographic techniques.Seven years after the Amoco Cadiz wreck, the oysters sampled in the AberBenoit and in the Baie de Morlaix (Carantec) are still polluted with residual aromatic hydrocarbons, corresponding, respectively, to about five and two times the values from an unpolluted site (Baie de Saint-Brieuc). Since petroleum residues are buried in anaerobic sediments, especially in the Abers areas, they are expected to persist and accordingly may still contaminate oysters for several more years.Depuration kinetics of contaminated flat and Pacific oysters from the heavily polluted zones (Abers, Carantec) were studied and compared with that of healthy oysters transplanted in the same areas. The decontamination rates of oysters after transfer to a clean environment were heavily dependent on the duration of oil exposure.Necrosis and inflammation were the most significant and noticeable changes occurring during the first months after the pollution. Atrophy of the gonadal cells was detected in the oysters from the Abers only during the first six months of the study. No neoplastic and parasitic diseases have been related to this oil spill.     

The effects of the ‘Braer’ oil spill, Shetland Isles, Scotland, on P4501A in fanned Atlantic salmon (Salmo salar) and the common dab (Limanda limanda)

This paper describes the response of two fish species, the Atlantic salmon (Salmo salar) and the common dab (Limanda limanda) to the oil spilled from the Braer tanker which grounded on the southern tip of Shetland, Scotland, on 5 January 1993. Both the immediate sublethal effects and the long-term response to the oil which accumulated in sediments around the Shetland Isles, Scotland, are examined. The primary response measured was the induction of detoxification enzymes and the relationship of the levels or activities of these enzymes to the concentration and distribution of aromatic hydrocarbons measured in sediments, water and fish. The results show that, immediately following the spill, there was a marked induction of Cytochrome P4501A enzymes in salmon, indicative of exposure to bioavailable aromatics. In dab there was evidence of induction at some sites in January 1993 immediately following the spill but, subsequently, no induction has been observed in fish caught in the vicinity of Shetland despite the very high concentrations of oil measured in sediments at some locations. This would indicate that the polyaromatic hydrocarbons in these sediments are unlikely to be directly bioavailable to fish. The effect of the oil spill on hepatic pathology in dab was investigated and shows that pathology predictive of neoplasia, such as basophilic foci and vacuolation, were observed in fish from the most contaminated sites in 1994 but not in 1993. The incidence of this pathology appeared to correlate with the degree of contamination at the sites, but additional studies are required to establish whether this was a consequence of the initial impact and exposure from waterborne oil at the time of the spill or whether it was due to the continued exposure to oil from the sediments.     

Bioavailability of heavy metals bound to sediments affected by a mining spill using Solea senegalensis and Scrobicularia plana

After the Aznalcóllar mining spill (April, 1998) different ecosystems along the Guadiamar river and the Guadalquivir estuary were impacted by high concentrations of metals. The concentration of metals Zn, Cd, Pb, Cu and As has been detected as a good tracer of the pathway of the toxic spill. Different individuals of the fish Solea senegalensis and the clam Scrobicularia plana were exposed to contaminated sediments after the accidental spill in the south-west Spain. The induction of metallothioneins in fish and the bioaccumulation of metals in clams were determined in different tissues to determine the biological adverse effect associated with the heavy metals from the spill. A relationship between the enrichment of metals in the tissues and the induction of metallothioneins was determined for those assays performed using contaminated sediments. The bioaccumulation of metals indicates that the risk associated with the spill should be continuously evaluated to establish the sublethal effects related to the accident.     

Effects of a crude oil and an oil dispersant (Corexit 9527) on populations of the littleneck clam (protothaca staminea)

Field and laboratory experiments were carried out to investigate the effects of Alberta crude oil and an oil dispersant (Corexit 9527) on the larval settlement, survival, siphon activities and behaviour of the littleneck clam (Protothaca staminea). Corexit 9527 was much more toxic than crude oil, and the highest toxicity was obtained when Corexit 9527 was mixed with crude oil. Siphon activities were impaired and abnormal behaviour was exhibited when adult clams were treated with 100 ppm Corexit 9527, 1000 ppm crude oil or a combination of both. Larval settlement was not affected when the substratum was treated with 1000ppm crude oil but was retarded when the substratum was treated with a mixture of 1000 ppm oil and 100 ppm Corexit 9527. Gas chromatograms also showed that the retention time and depth of penetration of hydrocarbons in the substratum was increased when Corexit 9527 was used with crude oil.     

Detection of the water-soluble fraction of crude oil by the blue crab, Callinectes sapidus

The ability of the blue crab, Callinectes sapidus, to detect petroleum hydrocarbons was measured with behavioural techniques. When presented with a water-soluble fraction of Prudhoe Bay crude oil, blue crabs abruptly changed antennular orientation, began rhythmic beating of the maxillipedal flagellae, and increased antennular flicking rate. The threshold concentration at which 50% of the crabs detected the water-soluble fraction was 2 × 10^?6 mg/litre. The blue crab apparently can readily detect petroleum hydrocarbons at concentrations found in chronically polluted areas as well as oil spill situations.     

Uptake of petroleum hydrocarbons by the Blue Mussel (Mytilus edulis L.) after Experimental Oiling and High Pressure, Hot Water Shore cleaning

High pressure, hot water shore cleaning after an oil spill will release high concentrations of petroleum hydrocarbons to ambient marine ecosystems. The immediate increase of hydrocarbons observed in blue mussels, Mytilus edulis, went from background concentrations of 40 μg/g to 657 μg/g and 533 μg/g at a distance of 3 and 8m respectively from the shore. After two weeks the accumulated oil had decreased by 20–45 %. In comparison natural surf and ice cleaning of shores will only produce a small increase in hydrocarbon concentrations. We recommend that high pressure, hot water cleaning is not used in areas where no special bird or wild life protection is needed.     

Hindcast of the Argo merchant spill using the uri oil spill fates model

A comprehensive oilspill fates model, developed by Cornillon and Spaulding at the University of Rhode Island, has been employed to hindcast the well-studied Argo Merchant spill. Using data collected during the spill to define the environmental conditions and the oil composition, properties and release rates, the oil spill fates model was used to predict the spatial and temporal distribution of spilled oil from December 16 to 31, 1976. Comparison of model predictions to observations for spilled oil mass balance, surface trajectories and oiled surface area was in general quite good. Improved predictions require a better definition of the wind and tidally-forced circulation in the vicinity of the wreck site, a more soundly based theory for oil entrainment or dispersion into the water column and an oil spreading theory which more accurately addresses the complex behaviour of a weathering multicomponent petroleum hydrocarbon.     

Induction of heat shock protein (hsp)60 in Isochrysis galbana exposed to sublethal preparations of dispersant and Prudhoe Bay crude oil

Adaptation to sublethal exposure to crude oil by phytoplankton is poorly understood. Use of chemical dispersants for oil spill remediation increases petroleum hydrocarbon concentrations in water, while exposing marine organisms to potentially toxic concentrations of dispersant. Heat shock proteins (hsps) have been found to serve as an adaptive and protective mechanism against environmental stresses. The objective of this project was to examine the induction of hsps in Isochrysis galbana, a golden-brown algae, following exposure to the water-accommodated fraction (WAF) of Prudhoe Bay crude oil (PBCO) and PBCO chemically dispersed with Corexit 9527® (dispersed oil: DO). Initial experiments using ³⁵S-labeled amino acids and 2-dimensional electrophoresis with subsequent western blotting identified and confirmed hsp60, a member of the chaperonin family of stress proteins, as being efficiently induced by heat shock in this species. One-dimensional SDS PAGE and western blotting, with hsp60 antibodies and chemiluminesence detection, were used to quantitate hsp60 following exposure to a range of environmental temperatures and concentrations of WAF and DO preparations. I. galbana cultured in 22 parts per thousand (‰) salinity showed a statistically significant increase (p<0.05) in hsp60 after exposure to 25, 30, and 35°C for 1 h compared to controls (20°C), while 34‰ cultures showed no significant increase in hsp60 with increasing temperature. Cultures (22 and 34‰) exposed to varying doses of WAF and DO exhibited statistically significant (p<0.05) increases in levels of hsp60 in all cases. A dose-related response was observed in all exposures (r²≥0.90) with the exception of 34‰ WAF. In independent exposures, a dose-related hsp60 response to naphthalene, a relatively abundant water-soluble aromatic hydrocarbon, was observed in both 22 and 34‰ cultures. Results of this study are consistent with previous studies in other species documenting increases in hsp60 levels with exposure to xenobiotics. Further studies are investigating the protective function of hsp60 against the toxic effects of exposure to WAF and DO preparations.     

Toxicity of sea-surface microlayer: Effects of hexane extract on baltic herring (Clupea harengus) and atlantic cod (Gadus morhua) embryos

This study was designed to determine whether contaminated sea-surface microlayer was toxic to marine fish embryos in its ntaive form and as a hexane extract. Developing embryos of Atlantic cod (Gadus morhua) and Baltic herring (Clupea harengus) were exposed to hexane extracts of sea-surface microlayer collected from five locations in the North Sea and Baltic Sea. Extracts from two of these locations produced significant embryos mortality as well as severe deformities in live hatched larvae. A control sample of bulk water collected from 20 cm under the surface and extracted in the same way produced no significant mortality or deformities. Significaant changes in timing of hatching were also observed in those samples which produced embryo toxic effects. A comparison of these data with those obtained from code embryos exposed to unextracted microlayer showed a similar biological effect with both unextracted samples and hexane extracts.Chemical analyses revealed the greatest biological effect in samples with petroleum hydrocarbon concentrations between 180 and > 200 μg liter⁻¹. The bulk water control had 1 μg liter⁻¹ while the three samples that showed no biological activity had 3 to 8 μg liter⁻¹ petroleum hydrocarbons. Phthalic acid esters were detected in four samples and chlorinated hydrocarbons in one, but could not be positively correlated with any of the toxic responses. No other chemical contaminants were detected in the five samples.The data presented here show that some sites contain sea-surface microlayer which can be toxic to marine fish embryos: that Baltic herring and Atlantic cod embryos respond similarly to the toxic effects of contaminated microlayer: and that unextracted microlayer and hexane extract of microlayer produce essentially the same toxic effect(s) if only organic contaminants are considered.     

Histochemistry as a tool for examining possible pathologic cause-and-effect relationships between heavy metal and inflammatory lesions in oysters, Crassostrea virginica

Unexplained inflammatory lesions have been seen on many occasions in oysters (Crassostrea virginica) that have been examined by histopathologic methods. Application of the Mallory's hematin and the rubeanic acid methods for demonstration of copper in histologic sections revealed strong staining reactions in granules of some, but not all, granular hemocytes. These cells were clearly involved as constituents of acute inflammatory lesions characterized by infiltration of arteries, occlusion of veins, and diapedesis through external epithelia of the mantle and gastrointestinal tract. Heavily contaminated sites (i.e. Raritan Bay, New Jersey) showed strong staining reactions in a large number of animals examined. Control sites such as Great Bay, New Jersey, showed much lighter reactions. Seasonal sampling in the New Jersey sites revealed the most intense involvement in the late summer with apparent decreasing staining intensities in the fall. Other species of bivalve mollusks (Mytilus edulis and Mercenaria mercenaria) only showed significant staining reactions for heavy metal in kidney tissues with no evidence that heavily contaminated sites had higher levels. These results constitute preliminary evidence of a cause-and-effect relationship between a heavy metal contaminant (most probably copper) and inflammatory lesions in the oyster.     

Effects of dietary and water-borne oil exposure on larval pacific herring (Clupea harengus pallasi)

Pacific herring (Clupea harengus pallasi) larvae were exposed either directly to the water-soluble fraction (WSF) of crude oil or indirectly via oil-contaminated prey (OCP) (Artemia salina) for <- 28 days, to determine the relative effects of diet and water as routes of contamination. Larvae were affected rapidly by 0·9 ppm WSF, which caused high larval mortality, reduced swimming ability and rapid reductions in feeding rates. Larval length was significantly reduced by 0·7 ppm WSF in 7 days and by 0·3 ppm WSF within 14 days. The WSF exposure also caused similar reductions in larval weights. Highly contaminated prey (6 ppm prey exposure) caused significant mortality, but surviving larvae appeared robust. The OCP did not affect swimming, feeding, or growth. Exposure of larvae was not significantly extended by OCP, which rapidly depurated WSF in clean water (98% in 1 day). Therefore, OCP is probably not an important source of low molecular weight petroleum contamination to larval fish in the marine environment.     

Effects of polynuclear aromatic hydrocarbons on fishes and shellfish: An overview of research in virginia

This paper discusses the following topics related to polynuclear aromatic hydrocarbon pollution in estuaries: (1) the use of oysters (Crassostrea virginica), hard clams (Mercenaria mercenaria) and brackish water clams (Rangia cuneata) in residue monitoring; (2) the effects of elevated PAH residues on oyster condition; (3) three years of field studies relating PAH sediment contamination to abnormalities in fishes; and (4) laboratory bioassays for effects and bioconcentration modeling. Oysters, hard clams and Rangia have been shown, from 3 years of field studies, to be good monitors of pollution inputs as one proceeds along salinity gradients from 25‰ to 0·5‰. Effects of increased body burdens of PAHs are shown by a lowering of the oysters' condition index, as measured by lipid levels. Fishes inhabiting the Elizabeth River, VA, which is highly contaminated with PAHs, have abnormalities (cataracts, skin lesions, abnormal fins, etc.). These abnormalities have higher incidence in regions of the river where the sediments are more heavily contaminated. Laboratory studies utilizing contaminated sediments have reproduced some of the abnormalities observed in the field. Bioconcentration of PAHs from sediments has been studied with oysters and hard clams. Oysters generally accumulate three times the body burdens of clams exposed to the same suspensions. A faster depuration rate (k₂) for clams appears to be responsible for the higher equilibrium body burden of oysters.     

消油剂处理120#燃料油对海水青鳉(Oryzias melastigma)胚胎抗氧化酶活性影响的研究

以海水青鳉(Oryzias melastigma)胚胎为研究对象,比较了120#燃料油分散液(water-accommodated fractions,WAFs)与乳化液(biologically enhanced water-accommodated fractions,BE-WAFs)的急性毒性效应,并研究了不同浓度(40、100、250 mg/L)下WAFs、BE-WAFs对胚胎内超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽硫转移酶(GST)活性的影响。结果表明:在受到石油烃的氧化胁迫后,海水青鳉胚胎内3种抗氧化酶活性变化明显。随着石油烃浓度的升高和暴露时间的延长,3种酶表现出程度不同的诱导效应和抑制效应。其中受石油烃污染影响最为明显的为SOD酶;而GST酶则对消油剂单独暴露表现较为敏感。实验证明,海水青鳉体内SOD酶活性对石油烃污染反应最为敏感,适合作为监测石油烃污染程度的生物标志物。     

The effects of petroleum hydrocarbons on lipid metabolism and energetics of larval development and metamorphosis in the american lobster (homarus americanus Milne Edwards)

In studies with larval stages of the American lobster (Homarus americanus) exposed to a sublethal concentration of South Louisiana crude oil, the normal patterns of lipid storage, utilization and synthesis during larval development and metamorphosis were altered with oil exposure. Increased rates of protein catabolism, delayed molting and reduced growth were also evident among oil-exposed lobster larvae and suggest either deficiency or immobilization of lipid reserves. From analyses of lipid class composition of control and oil-exposed lobster larvae, it is evident that oil-exposed larvae have consistently lower levels of triacylglycerols (the major energy store) and higher levels of sterols than control larvae. Decreased ratios of non-essential fatty acids (both saturated and unsaturated) and increased ratios of essential fatty acids were detected in the neutral lipid pools of oil-exposed larvae that suggest both decreased storage of fatty acids in energy reserves and decreased mobilization of essential fatty acids into phospholipid pools. The fatty acid distribution in the phospholipid pools of oil-exposed larvae, however, varied little from control values and reflects a tendency to preserve the integrity of these structural lipids. Increases in sterol levels may be related to decreased mobilization of cholesterol or other sterols and/or transformation of cholesterol into metabolic pathways including the biosynthesis of ecdysterone.Oil-exposed animals contained trace quantities of benzene, thiophene, toluene, alkylcyclohexane and alkylbenzenes, detected with GC-MS analyses. No naphthalene, dimethylnaphthalene, higher molecular weight aromatics or their phenolic derivatives were detected; their absence may be related to either the detection limits of the methodology used or extremely rapid metabolism and turnover of these components. Alterations in lipid metabolism may account for the developmental and energetic abnormalities observed in marine crustaceans. It cannot be ruled out, however, that decreased lipid utilization, synthesis and mobilization may be a defense mechanism against incorporating lipophilic components of petroleum hydrocarbons in metabolic pathways, and that disruption in energetics and development is a consequence of the reduction in energy available for growth and molting.     

Use of the Aurelia Metamorphosis Test System to detect subtle effects of selected hydrocarbons and petroleum oil

The Aurelia Metamorphosis Test System was devised to use simple Aurelia polyps as indicator organisms for the detection of subtle effects of petroleum-related hydrocarbons and of Alaskan crude petroleum oil (ACPO) on developmental processes involved in strobilation initiation, differentiation of ephyrae, and genetic modification expressed through cloning. Compounds tested were ACPO, aniline, anthracene, benzanthracene, benzene, benzo[a]pyrene, biphenyl, cresol, naphthalene, perylene, phenol and pyrene. Gross morphology of the ephyrae (patterning, numbers and appearance of structures), statolith and mematocyst numbers, and pulsing behavior were recorded using the light microscope. Selected ephyrae with teratological malformations were studied with the scanning electron microscope and swimming behavior was observed visually. The variety of subtle developmental, behavioral and genetic effects due to marine hydrocarbons detected using the Aurelia Metamorphosis Test System demonstrated that this test system could be used as an early warning system to detect subtle effects of other marine pollutants.     

Candidate biomarker discovery in plasma of juvenile cod (Gadus morhua) exposed to crude North Sea oil, alkyl phenols and polycyclic aromatic hydrocarbons (PAHs)

In this study we have investigated protein changes in plasma of juvenile Atlantic cod (Gadus morhua) induced by crude North Sea oil and North Sea oil spiked with alkyl phenols and polycyclic aromatic hydrocarbons, a surrogate produced water composition. Using a proteomic approach, we identified 137 differentially expressed proteins at different levels of crude oil exposure. Many of the induced protein changes occurred at low levels of exposure. The results obtained with protein expression profiles after exposure to oil and surrogate produced water indicate effects on fibrinolysis and the complement cascade, the immune system, fertility-linked proteins, bone resorption, fatty acid metabolism as well as increased oxidative stress, impaired cell mobility and increased levels of proteins associated with apoptosis. Although the number of individuals and samples in this study is limited within each treatment group, the protein changes observed in this study represent a first screening for potential biomarker candidates in cod plasma reflecting potential effects of crude oil and produced water exposure on fish.     

A chemical investigation of the transport and fate of petroleum hydrocarbons in littoral and benthic environments: The TSESIS oil spill

The fate of saturated and aromatic hydrocarbons discharged into the coastal Baltic Sea environment from the TSESIS oil spill has been studied in the acute and postacute (one year) phases of the spill. Periodic samples of Mytilus edulis (mussels) from eight littoral zone stations and Macoma balthica from nine soft bottom stations were obtained as well as sediment trap samples and surface sediment samples. Glass capillary gas chromatography and gas chromatographic mass spectrometry were used as the analytical tools to determine saturated and aromatic hydrocarbon composition and concentrations in these samples.Sediment trap samples indicated that sizable quantities of chemically and microbially weathered oil were sedimented, and available for benthic uptake shortly after the spill. After initial uptake of sedimented oil (500 to 1000 μ/g dry weight), Macoma populations appear to have begun slow depuration through the first winter after the spill, but TSESIS oil was again introduced to the benthic stations studied during the following summer. Mytilus populations in the region were severely impacted by the oil. Initial depuration of spilled oil during the first month was rapid and nearly complete at all but the most heavily impacted stations one year after the spill. The post-spill depuration of assimilated hydrocarbons was characterised by a relative retention of alkylated dibenzothiophenes and alkylated phenanthrenes compared to their unsubstituted parent compounds and compared with the entire homologous naphthalene series.These data suggest that petroleum hydrocarbons from the TSESIS spill have become a chronic source of degraded saturated and aromatic hydrocarbons to the soft bottom benthic communities. Petroleum hydrocarbons in the benthic environment from this spill appear to reside in the difficulty sampled and mobile flucculent layer at the sediment/water interface and may affect epifaunal communities for an extended period of time.     

Exposure to hydrocarbons 10 years after the Exxon Valdez oil spill: evidence from cytochrome P4501A expression and biliary FACs in nearshore demersal fishes

Three biomarkers of hydrocarbon exposure, CYP1A in liver vascular endothelium, liver ethoxyresorufin O-deethylase (EROD), and biliary fluorescent aromatic compounds (FACs), were examined in the nearshore fishes, masked greenling (Hexagrammos octogrammus) and crescent gunnel (Pholis laeta), collected in Prince William Sound, Alaska, 7-10 years after the Exxon Valdez oil spill (EVOS). All biomarkers were elevated in fish collected from sites originally oiled, in comparison to fish from unoiled sites. In 1998, endothelial CYP1A in masked greenling from sites that were heavily oiled in 1989 was significantly higher than in fish collected outside the spill trajectory. In 1999, fishes collected from sites adjacent to intertidal mussel beds containing lingering Exxon Valdez oil had elevated endothelial CYP1A and EROD, and high concentrations of biliary FACs. Fishes from sites near unoiled mussel beds, but within the original spill trajectory, also showed evidence of hydrocarbon exposure, although there were no correlations between sediment petroleum hydrocarbon and any of the biomarkers. Our data show that 10 years after the spill, nearshore fishes within the original spill zone were still exposed to residual EVOS hydrocarbons.     

基于特征比值的原油中PAHs标志物风化规律研究

以渤海某原油作为研究对象进行综合模拟风化实验, 探讨了原油中五类PAHs组分的风化规律并筛选出其中稳定的诊断比值。结果表明: 经过50d风化, 原油中PAHs的分布已经发生了较大的改变, 其中萘系类损失最为严重, 菲系列所占的比例有所提高, 二苯并噻吩、屈、芴系列则保持相对稳定, 这为进一步筛选用于溢油来源鉴别的新诊断比值参数提供了一定依据; 经风化检验, 现有常用6种PAHs诊断比值在风化50d后较稳定, 可用于风化溢油的鉴别; 所选取新诊断比值中, 菲和屈系列、烷基取代二苯并噻吩类、烷基取代芴类抗风化能力较强, 共筛选出34种诊断比值可作为溢油鉴别的有效指标。     

多环芳烃油指纹应用于船舶溢油鉴别研究

溢油种类主要包括船舶燃料油和原油,二者性质的差异决定了鉴别方法也相应不同,寻求适合于船舶溢油的鉴别方法具有重要意义。在使用柱色谱层析方法对样品进行分离前处理的基础上,以气相色谱/质谱方法(GC-MS)为主要分析手段,对溢油样品和可疑船舶溢油源样品的多环芳烃油指纹特征进行对比,并在多环芳烃油指纹参数的基础上进一步进行多环芳烃内组成三角图分布特征与聚类分析研究,成功为珠江口水域某船舶溢油事故追踪到肇事溢油源。结果表明:取自丁船的油样和现场溢油样芳烃油指纹特征最为相近,是此次溢油事故的溢油源。受风化作用后的船舶燃料油中饱和烃类化合物数量稀少,且含量极低,不适合用于溢油鉴别,而多环芳烃类化合物较饱和烃类化合物而言具有含量高、种类丰富的特点,是该类溢油鉴别的主要油指纹依据。使用油指纹参数进行可疑溢油源识别时,充分考虑油品中有机分子所受风化影响程度的不同是风化条件下溢油鉴定的关键。因此,多环芳烃油指纹可以有效应用于船舶燃料油溢油的鉴别。