Characteristics of dispersions and water-soluble extracts of crude and refined oils and their toxicity to estuarine crustaceans and fish

The quantitative hydrocarbon composition and behavior in seawater of water-soluble fractions (WSF) and oil-in-water dispersions (OWD) of 4 oils was investigated. Two crude oils, South Louisiana crude and Kuwait crude, and two refined oils, No. 2 fuel oil and bunker C residual oil, were used in these investigations. The WSFs of the crude oils had higher total oil-hydrocarbon concentrations and were richer in light aliphatics and single-ring aromatics than were the WSFs of the refined oils. The WSFs of the refined oils contained significantly higher concentrations of naphthalenes than did those of the crudes. The hydrocarbon composition of the aqueous phase of OWDs closely resembled that of the parent oils. Gentle aeration of the OWDs resulted in a loss of 80 to 90% of the aqueous hydrocarbons in 24 h. Alkanes disappeared from the dispersions more rapidly than aromatics. The WSFs and OWDs of the refined oils were considerably more toxic to the 6 test species than were those of the crude oils. The test species can be ranked according to increasing sensitivity to oil as follows: Cyprinodon variegatus, Menidia beryllina, Fundulus similus, Penaeus aztecus postlarvae, Palaemonetes pugio and Mysidopsis almyra. The results of this investigation are discussed in relation to the potential impacts of oil spills on the marine and estuarine environments.     

天津地区表层土壤和河流沉积物中甾烷与五环三萜烷的分布特征及污染源分析

分析了天津地区表层土壤和具有代表性的河流沉积物中甾烷和五环三萜烷系列化合物的组成与分布特征，讨论了这类化合物的来源及环境意义。分析表明，在天津地区不同环境功能区表层土壤和河流沉积物中均检测到了甾烷和五环三萜烷系列化合物，但样品之间其含量存在明显的差别；表层土壤与河流沉积物中甾烷、五环三萜烷化合物的组成与石油中的基本一致，样品间这类化合物在饱和烃中的相对含量与正烷烃CPI之间有较好的负相关关系，表明它们主要来源于石油及副产品，可以根据样品中甾、萜化合物与正构烷烃的比值来反映饱和烃中石油烃污染源的贡献。论文根据样品中甾烷、五环三萜烷化合物与正构烷烃的比值，并结合正烷烃CPI参数初步分析了天津地区不同环境功能区表层土壤和河流沉积物中饱和烃污染物的污染源。     

The effects of a no. 2 fuel oil and two crude oils on the growth and photosynthesis of microalgae

Seawater, when equilibrated with a sample of No. 2 fuel oil, becomes toxic in varying degrees to growth of representative types of microalgae, two blue-greens, a diatom, two greens, and a dinoflagellate. For a sensitive organism such as Thalassiosira pseudonana, Strain 3H, 5 ml of seawater equilibrated with fuel oil (containing 15 mg/l of organics) in 20 ml of growth medium is lethal, or roughly in the range of 40 to 400 ppb if the toxic material(s) constitute 1 to 10% water also immediately stops photosynthesis in organism 3H. For other microalgae tested e.g. 580 (a green alga) and PR-6 (a blue-green alga), similar effects on growth and photosynthesis were found, but required higher concentrations of the oil-equilibrated seawater. Water solubles from Kuwait or Southern Louisiana crude oils (when the straight crude oil was equilibrated 1:8 with seawater) were not toxic; however, specific fractions obtained by distillation did show some water-soluble toxicity. Growth experimetns in open or closed growth systems revealed that most organisms were inhibited by varying amounts of these two crude oils when in direct contact with them. Organism 580 would not grow above 5 μl of Southern Louisiana/25 ml of medium, or 10 μl of Kuwait/25 ml of medium (oil in direct contact with algae). With both the seawater equilibrated with fuel oil and the crude oils, the toxic activity is mainly localized in medium and higher boiling fractions derived from distillation cuts from these materials.     

Effect of microbial seeding of crude oil in seawater in a model system

The adding of a mixed culture of marine hydrocarbon-degrading bacteria to petroleum polluted seawater did not increase the hydrocarbon degradation capability of the water. All the strains of the mixed culture disappeared from the dominant microflora while the autochthonous bacteria showed a capacity for adaptation to petroleum degradation about four days after the oil spill. This confirms the advantage of natural bacterial communities. The oil spilled on the sea surface evolved very quickly under the influence of abiotic factors such as sunlight. Aromatic petroleum fractions were oxidized and polymerized. This chemical evolution causes a drastic decrease of the biodegradability of crude oil.     

Responses of Mytilus edulis on exposure to the water-accommodated fraction of North Sea oil

Individuals of Mytilus edulis L., collected from the Erme estuary (S.W. England) in 1978, were exposed to low concentrations (7 to 68 g l^-1) of the water-accommodated fraction (WAF) of North Sea crude oil. The pattern of accumulation of petroleum hydrocarbons in the body tissues was affected by the presence of algal food cells, the period of exposure, the hydrocarbon concentration in seawater, the type of body tissue and the nature of the hydrocarbon. Many physiological responses (e.g. rates of oxygen consumption, feeding, excretion, and scope for growth), cellular responses (e.g. lysosomal latency and digestive cell size) and biochemical responses (e.g. specific activities of several enzymes) were significantly altered by short-term (4 wk) and/or long-term (5 mo) exposure to WAF. Stress indices such as scope for growth and lysosomal latency were negatively correlated with tissue aromatic hydrocarbons.     

Hydrocarbons characterization in coastal sediments of the Argentine Patagonia using the nuclear magnetic resonance (NMR) spectroscopy

Hydrocarbons in Patagonian sediments were analyzed to evaluate their biogenic or petrogenic origins. Structural determination by ¹H-NMR spectrometry and principal component analysis (PCA) show that the first two principal components accounted for more than 75.7% of the variance. The results indicate that biogenic hydrocarbons have mainly linear hydrocarbon chains and polar groups (high H₂, H₄, H₅, and H_COOH), while petrogenic hydrocarbons show more branched chains and mainly mono-aromatic components, e.g., crude oil (high H₁, H_1A, and H₃) or mainly poly-aromatic components (high H₄, and H_2A) in weathering oil.     

流沙湾海水中石油烃的时空分布特征研究

于2008年2月（冬季）、5月（春季）、8月（夏季）和11月（秋季）对流沙湾进行了4次采样考察,研究分析了流沙湾表层海水中石油烃质量浓度的平面分布和季节变化特征。结果表明,在2008年度,流沙湾表层海水石油烃的质量浓度为0~1.930 mg.L-1,平均值为0.080 mg.L-1,季节差异比较明显,呈春、冬、夏、秋季依次减小的变化趋势,冬、春季节海水呈现不同程度石油污染,夏、秋季节属Ⅰ、Ⅱ级水质,整个流沙湾海域表层海水石油烃的平面分布相对比较均匀。在内外湾分布上,冬、春、秋季节外湾大于内湾,而夏季节内湾大于外湾。流沙湾的水产养殖活动是其海水石油烃时空分布的主要影响因素。     

Short-term effect of microencapsulated hydrocarbons on shell growth of Mytilus edulis

Two North Sea crude oils, weathered crude oil, n-alkanes, medical liquid paraffin and a fish oil were microencapsulated and added to seawater. This gave a medium containing both water soluble fractions (WSF) and oil particles, and thus simulated the conditions in natural seawater contaminated with oil. The microencapsulated oils were fed to Mytilus edulis L. in different concentrations, and the growth in terms of shell length of the mussels was measured in intervals of 24 to 48 h for 4 to 12d. With the crude oilss A and B, weathered crude oil and n-alkanes at concentrations1 mg l^-1, shell growth rate decreased rapidly compared to controls. With liquid paraffin at levels of 1 to 12 mg l^-1, a small but significant negative effect on shell growth occurred after 5 d of exposure. Exposure to fish oil at 1 and 4 mg l^-1 gave no significant reduction in growth rate. The toxicity of the different hydrocarbons was not related to their content of aromatic fractions. Crude oil B was tested at concentrations ranging from 0.12 to 12 mg l^-1. At 0.12 mg l^-1 shell growth was not significantly different from the control, while at 0.25 and 0.50 mg l^-1 a temporary and significant stimulation of growth was observed. The product concentration (C)xresponse (R) shows a linear regression on exposure time (t). The regression model CxR=75-0.18t is used to estimate EC-values (effect of a given concentration) for given response levels. The model gives a very good fit to observed data.     

石油污染与微生物群落结构的相互影响

从两种土壤中分别分离出石油烃降解菌,并从中筛选出6株石油烃高效降解菌A1、A2、A6、A8和B2及B5,然后将各菌株鉴定至属,分别为A1假单胞菌属、A2鞘氨醇单胞菌属、A6微球菌属、A8节杆菌属、B2不动杆菌属和B5诺卡氏菌属。另外对比分析了单菌株及不同菌株重组对不同石油烃组分的利用情况,结果发现,从不同石油污染的土壤中分离到的菌株对石油烃组分的利用能力不同,从胜利原油污染的土壤中分离到的菌株A1、A2、A6和A8对石油烃组分的利用范围窄,主要利用饱和烃组分;而从经芳香烃驯化过的土壤中分离到的菌株B2及B5对石油烃利用组分的利用范围较宽,能同时利用饱和烃和芳香烃组分。     

Accumulation,fractionation and release of oil by the intertidal clam Macoma balthica

The bivalve mollusc Macoma balthica accumulated hydrocarbons during 180 days of continuous exposure to Prudhoe Bay crude oil in seawater dispersions with nominal concentrations of 0.03, 0.3 and 3.0 mg l^-1. The mollusc's ability to concentrate oil from seawater increased with decreasing oil-in-water concentration. Decreases in oil burden began after 30 to 120 days (depending on the oil concentration) and continued for at least 60 days after exposure to oil ceased. Aliphatic and aromatic hydrocarbons were fractionated in markedly different ways by the bivalve. Branched and cyclic aliphatics in the molecular weight-range dodecane through hexadecane were preferentially retained over straightchain and their higher homologs. Larger and more substituted aromatic compounds were selectively concentrated. There appeared to be no selective concentration of aromatic sulphur compounds.Please address requests for reprints to Dr. D. G. Shaw at the Institute of Marine Science     

重质原油和轻质原油水溶性组分对紫贻贝(Mytilus edulis)毒理效应的研究

近年来,海洋石油开采与运输泄漏、石油及产品离岸排放等事故逐年增多,对近海海域生态环境产生了巨大破坏。为探究重质原油和轻质原油对紫贻贝(Mytilus edulis)毒理效应,测定了不同浓度重质原油和轻质原油暴露下紫贻贝鳃和外套膜中过氧化氢酶(CAT)和超氧化物歧化酶(SOD)的活性及体内石油烃的含量变化。结果显示,石油污染暴露下,紫贻贝腮和外套膜中CAT和SOD活性变化明显,与暴露浓度和暴露时间有关。CAT活性在轻质原油组随着浓度的增大,呈现下降趋势,且随着暴露时间的延长呈现先下降后上升趋势;在重质原油暴露组,随着浓度的增大,呈现先上升后下降趋势,且随着暴露时间的延长呈现先下降后上升趋势。SOD活性在轻质原油与重质原油暴露组,随着浓度的增大,呈现下降趋势,且随着暴露时间的延长呈现上升趋势并存在一定的剂量-效应关系。重质原油和轻质原油暴露168 h后紫贻贝体内总石油烃含量呈线性递增,生物富集系数(BCF)随着暴露浓度的增加不断减小并最终趋于平稳。结果表明,以紫贻贝腮和外套膜中SOD和CAT活性作为石油烃污染的生物标志物具有一定应用前景,紫贻贝对原油溶液中石油烃的生物富集作用可用来判断污染原油的来源和性质。     

0~#柴油和流花原油对斑节对虾(Penaeus monodon)幼体的急性毒性效应

溢油污染导致的原油和燃料油入海,会对海洋生物的生长发育过程产生影响。为研究溢油污染对海洋虾类的毒性效应,以斑节对虾(Penaeus monodon)为研究对象,比较了不同浓度0#柴油和南海流花原油(LH原油)乳化液对斑节对虾不同发育阶段幼体的急性毒性效应。结果表明,3.59 mg·L~(-1)0#柴油和0.77 mg·L~(-1)LH原油乳化液可以显著降低斑节对虾无节幼体变态率(P0.05),且对无节幼体变态具有延迟效应。较之0#柴油,LH原油乳化液对斑节对虾无节幼体发育的影响更为明显。0#柴油对斑节对虾无节幼体、蚤状幼体、糠虾和仔虾的48或96小时半致死浓度(48 h/96 h-LC50)分别为0.55 mg·L~(-1)、0.42 mg·L~(-1)、0.95 mg·L~(-1)和1.09 mg·L~(-1),其对应的安全浓度分别为0.05 mg·L~(-1)、0.04 mg·L~(-1)、0.10 mg·L~(-1)和0.11 mg·L~(-1);LH原油对上述幼体的48 h/96 h-LC50则依次为0.62 mg·L~(-1)、0.51 mg·L~(-1)、1.05 mg·L~(-1)和1.42 mg·L~(-1),对应的安全浓度分别为0.06 mg·L~(-1)、0.05mg·L~(-1)、0.11 mg·L~(-1)和0.14 mg·L~(-1)。斑节对虾不同发育阶段幼体对0#柴油和LH原油的耐受力依次为:仔虾糠虾无节幼体蚤状幼体,0#柴油和LH原油乳化液对斑节对虾的毒性大小为0#柴油LH原油。上述结果为深入研究石油类污染对海洋生物的毒性效应提供了基础数据和理论依据。     

Sublethal effects of chronic oil exposure on the intertidal clam Macoma balthica

When exposed to Prudhoe Bay crude oil in flowing seawater for 180 days, the small intertidal clam Macoma balthica showed behavioral, physical, physiological and biochemical changes. At a high concentration of oil in seawater (3.0 mg l^-1) burrowing rate decreased, respiration rate increased, growth was inhibited, and very high mortalities resulted. The lowest concentration of oil in seawater (0.03 mg l^-1) inhibited growth and caused reabsorption of gametes. One group of adverse oil effects which was related to sluggishness and disorientation of the clams appeared after a week of exposure to oil; another group related to a negative energy balance was not observed until 60 days. We conclude that chronic exposure of M. balthica to oil-in-seawater concentrations even as low as 0.03 mg l^-1 will, in time, lead to population decreases.Please address requests for reprints to Dr. D. G. Shaw at the Institute of Marine Science     

Bioavailability of a hydrocarbon from water and sediment to the marine worm Arenicola marina

The uptake, accumulation and depuration of a labelled hydrocarbon (¹⁴C-1-naphthalene) by the marine annelid Arenicola marina were studied. Naphthalene was chosen because it is an important component of oil and is known to be toxic to marine organisms. The bioavailability of the hydrocarbon to the worm from contaminated sediment is examined and the results discussed with reference to the rehabilitation of oiled environments, as A. marina is thought to be capable, in high numbers, of greatly reworking sediments.     

Possible Use of Rna:Dna Ratio for Detecting Oil Induced Disturbance: A Field Report

The effects of oil contamination on the metabolic state of suspended participate and sedimented organic matter were investigated through the analysis of the RNA:DNA ratio on samples collected from January 1991 to January 1992 in the Ligurian Sea (northwestern Mediterranean). the ratio of RNA to DNA appeared to be highly sensitive to hydrocarbon stress since the oil disturbance resulted in an evident decline in the RNA:DNA ratio relative to the pre-pollution conditions. Such a pattern was unusual since it occurred in a period when we would expect a seasonal increase in bacterial activity. Differences are seen when comparing the temporal trend of DNA in suspended particulates and in sediments. However, the effects of oil on the RNA:DNA ratio were similar. Oiled samples showed a RNA:DNA ratio lower than 1 whereas RNA concentrations exceeded DNA content in undisturbed samples. Bacterial DNA accounted for a significant fraction (average 26%) of total DNA in sediments. It is likely that changes in RNA:DNA ratio in oiled sediments are due to the disturbance of bacterial communities. Consistent with the meiofaunal response to oil contamination, the hydrocarbons had only temporary and limited effects and RNA:DNA values recovered to pre-pollution conditions after a few weeks. the possible use of the RNA:DNA ratio in pollution monitoring is discussed.     

Effects of exposure to sublethal concentrations of crude oil on the copepod Centropages hamatus

Female copepods of the species Centropages hamatus show decreased ingestion rates and decreased egg viability when exposed to crude oil/seawater dispersions having crude oil concentrations of 10–80 ppb. However, rates of egg production were not significantly affected by these exposure levels. In addition, we found no evidence for accumulation of petroleum hydrocarbons by copepods exposed to 200 ppb of South Louisiana crude oil. The results imply that biosynthetic pathways involved in oogenesis may be influenced by sublethal concentrations of crude oil or that petroleum hydrocarbons directly affect the viability of eggs. Recruitment into field populations of copepods could be severely reduced as a consequence of exposure to low levels of physically dispersed crude oil.     

Experimental field study of the effects of crude oil,drill cuttings and natural biodeposits on microphyto- and macrozoobenthic communities in a Mediterranean area

Effects of an experimental pollution by neogenous and fossil organic matter on microphyto- and macrozoobenthic communities were studied in situ over a 1 yr period (July 1989 to May 1990) in a shallow microtidal bay (Gulf of Fos, south coast of France). Three experimental enclosures of 1 m² non-defaunated sediments were covered with 1 cm of polluted defaunated sediments. The sediment in one enclosure contained natural biodeposits with a high organic matter content (BD), that in a second enclosure contained Arabian light crude oil (BAL), and the sediment in the third enclosure contained diesel oil-based cuttings (CUT). Pollution by contaminants did not prevent microphytobenthos from colouizing sediments. Population changes over time were quite similar in all enclosures, except in CUT, where a four times higher chlorophyll a content appeared to be related to a decreased number of grazers and consequently lower grazing rates of animals. Toxicity to the fauna was immediate in the case of BAL and occurred within 3 mo at CUT. Opportunistic species settled in all contaminated sediments; this occurred quite rapidly in BD and BAL which recovered within 3 mo to levels comparable with control sediments. In CUT, natural populations had not recovered after 1 yr, whereas a quasi-monospecific population of Capitella capitata was still present. During the first 3 mo, the oxygen demand of the sediment was higher in oil-contaminated sediments than in controls. On the whole, the changes in fluxes and organism assemblages in our weakly tidal area appear to be consistent with other findings in macrotidal seas.     

Colonization of meiobenthos in oil-contaminated subtidal sands in the lower Chesapeake Bay

In-situ manipulative experiments were conducted over a 3-month period (May–August 1980) to examine the rate at which meiobenthos colonizes oiled and untreated azoic fine sands at a shallow subtidal site in the lower York River, Virginia. Three concentrations of fresh Prudhoe Bay crude oil were added to sediments: 100, 2 500 and 10000 mg oil kg^-1 dry wt sediment. Untreated azoic and natural sediments served as controls. Within 16 d, meiofauna densities in all treatments were comparable to natural populations in surface oxidized sediments, but densities fluctuated greatly during the remainder of the sampling period. Nematodes slowly colonized the subsurface anoxic sediments below the redox potential discontinuity (RPD); some less common species did not significantly recover below the RPD in the two more heavily oiled treatments. Analysis of nematode community composition by reciprocal averaging ordination and numerical classification revealed generally lower abundances, but no distinct differences, in species composition in the oiled substrates as compared to untreated and natural community controls. Ordination of sequential samples suggested that the nematode species assemblages in the untreated controls fully recovered from these small-scale disturbances by 90 d. Life history characteritics and frequent tidal transport combine to make estuarine meiobenthos highly resilient following disturbance. Contrary to prior recolonization studies, a successional sequence was found for the colonizing nematodes which may be analogous to models of macrobenthic colonization (e.g. McCall, 1975). The comesomatid nematode Sabatieria pulchra, which is frequently dominant in polluted sediments, colonized relatively late in the experiment. Consequently, stress resistance and resilience may not be as coincident in meiofauna as in macrofauna because of differences in factors affecting their dispersal.     

Hydrocarbons contamination and microbial degradation in mangrove sediments of the Niger Delta region (Nigeria)

The distribution of aliphatic hydrocarbons and polycyclic aromatic hydrocarbons (PAHs), and their degradability by bacteria in epipelic and benthic sediments from Qua Iboe Estuary mangrove ecosystem and associated creeks were investigated. The research findings revealed that total aliphatic hydrocarbons in sediments ranged from 16.82 mg·kg ^?1 to 210 mg kg ^?1, wheras total PAHs ranged from 6.30 to 35.55 mg·kg ^?1 dry weight of sediment. Low molecular mass (i.e. the 2–3-ring) PAHs were predominant in almost all the sampling points, whereas the higher molecular masses (4-, 5- and 6-ring PAHs) had the lowest concentrations. In general, the sediment samples ES ₂ (39.7%), ES ₃ (24.8%), BS ₁ (46.7%), BS ₂ (49.9%) and BS ₃ (44.2%) showed<50% contributions of Σ combustion–derived PAH (COMP-PAH) concentration to the Σ PAH concentrations, whereas ES ₁ (57%) contained>50% of COMP-PAHs. Our results have also shown that many mangrove bacteria have strong capacity to utilise Qua Iboe Light (QL) crude oil as the sole source of carbon and energy, while lower number of bacterial species including Bacillus sp., Micrococcus sp., Pseudomonas aeruginosa, Alcaligenes sp. and Flavobacterium sp. exhibited detectable PAHs degradability; and as such may serve as efficient degraders of QL crude oil contamination of mangrove ecosystem.     

Chemical characterization of a water soluble fraction (WSF) of crude oil

Very polar oil compounds are predominant in the resulting water soluble fraction (WSF) of crude oil, in agreement with their high solubility in seawater. Non‐volatile hydrocarbons represent only a low proportion of total soluble extracts.

The experimental process we have achieved allows us to obtain quantitative and reproducible soluble oil extracts that could be used in ecotoxicological tests and for the study and the characterization of marine oil pollution.