Uptake and Dupuration of Petroleum Hydrocarbons By the Backwater Clam, Meretrix Casta Chemnitz

Meretrix casta were experimentally exposed to water soluble fractions of refined and crude oil and their rate of accumulation of the petroleum hydrocarbons including total and individual aromatics viz., naphthalene, methylnaphthalene and dimethylnaphthalene, was studied. Subsequent transfer to clean waters in the laboratory and field resulted in depuration of the accumulated hydrocarbons from tissues. in general, the rate of discharge was found to be dependent on the concentration that the animals had been earlier exposed to and also the alkylation of aromatic hydrocarbons. Occasional increases were observed in the hydrocarbon values of clams which were placed in the field, compared to their laboratory counterparts, suggesting an intermittent source of petroleum hydrocarbon input into the environment. the influence on the rate of uptake, release and retention of hydrocarbons in the clams is discussed.     

The persistence of chronically accumulated hydrocarbons in the hard shell clam Mercenaria mercenaria

Hard shell clams, Mercenaria mercenaria, from the chronically polluted environment of the Providence River, USA, were transferred to a clean laboratory system located in lower Narragansett Bay, Rhode Island. The hydrocarbon contents of these transplanted clams were monitored for 120 days after transfer. After this period only slight depuration had occurred. A 41.9 g g wet weight^-1 average initial hydrocarbon burden decreased to 29.3 g g^-1, a decrease that became statistically significant (P<0.1) only after 120 days. At most, only 30% of the clams' hydrocarbons were lost. These findings indicate that the duration of the exposure period and the chemical nature of the source hydrocarbons are both instrumental in determining the persistence of these compounds in filter-feeding bivalves. Chronically accumulated petroleum hydrocarbons are strongly retained by the organisms and are only very slowly depurated.     

植物-微生物修复石油烃污染土壤与根际微生态环境变化

石油烃作为一类持久性难降解有机污染物对土壤环境质量产生严重的危害。以天津大港油田原油污染土壤中筛选出的耐低温高效石油烃降解菌为靶细胞,以小麦、紫花苜蓿作为供试植物,利用盆栽试验,对植物-外源菌协同修复体系中的脱氢酶活性和土壤微生物多样性进行研究,分析其变化及其与石油烃降解率的关系。结果表明植物-微生物协同修复对石油烃具有较好的降解能力,其中小麦-固定化外源菌组具有最高的降解率,石油烃含量从最初的30 600 mg獉kg-1下降为24 300 mg獉kg-1,降解率为20.6%,并且其试验后期石油烃的降解率最大,远远高于其他时期,表现出良好的修复潜力。外源菌投加的初始阶段会迅速提高脱氢酶活性,然而这种影响随着降解时间延长而逐渐减弱。初期脱氢酶活性与总石油烃的降解存在较好的相关性,脱氢酶活性可以在一定程度上表征土壤石油烃的降解情况。微生物多样性与总石油烃降解也存在一定的相关性。     

石油烃对栉孔扇贝血淋巴细胞DNA损伤的初步研究

为研究石油烃对海洋生物的毒性效应,将栉孔扇贝(Chlamys farreri)暴露于0.08、0.21和0.88mg·L-1石油烃中,采用单细胞凝胶电泳实验(彗星实验)技术检测不同暴露时间扇贝血淋巴细胞的DNA损伤程度,对照组中石油烃背景浓度为0.04mg·L-1。结果显示,低浓度(0.08mg·L-1)的石油烃短期(<7d)内即可导致栉孔扇贝血淋巴细胞的DNA损伤,并且随石油烃浓度的增大和暴露时间的延长,DNA损伤程度增加,石油烃浓度达0.88mg·L-1时,DNA损伤程度已非常严重。3d恢复实验后,各浓度组DNA损伤又均有不同程度的恢复。研究表明,彗星实验是检测石油烃对海洋贝类DNA损伤的一种有效手段,贝类血淋巴细胞DNA损伤有望成为石油烃污染的一种生物标志物,用于海洋污染的早期预警监测。     

Accumulation,release and retention of petroleum hydrocarbons by the oyster Crassostrea virginica

Two Crassostrea virginica populations, differing in fat content, were experimentally exposed to a complex petroleum-hydrocarbon fraction. The hydrocarbons in this mixture were accumulated by both groups of oysters, and their lipid content, as well as the concentration of hydrocarbon in the water, were found to affect the rate and extent of accumulation. Hydrocarbons accumulated were rapidly, although incompletely, discharged when the oysters were transferred to an uncontaminated system. Amounts of hydrocarbons discharged and amounts retained after discharge are probably related to the level of contamination. The data can be interpreted as indicating that equilibration and the occurrence of multiple compartments where hydrocarbons can reside are factors involved in the uptake and retention of nonbiogenic hydrocarbons by oysters. The petroleum hydrocarbons contained in the oysters differed from the contaminating oil by displaying a greater aromatic content. In addition, gas-liquid chromatograms of aliphatic fractions of the hydrocarbons in the oysters rapidly showed a degraded appearance; the possibility that the oysters themselves are modifying the oil cannot be excluded.Contribution No. 3098 from the Woods Hole Oceanographic Institution.     

Siphon nipping facilitates lethal predation in the clam Mesodesma mactroides (Reeve, 1854) (Mollusca: Bivalva)

In soft sediment marine communities, fishes frequently bite off extended siphons of buried clams; the consequential shortening of the siphon is known to reduce burial depth of the clams, secondarily increasing their vulnerability to lethal excavating predators. In this study, siphon nipping on the yellow clam, Mesodesma mactroides, was simulated by removing the top 6.6–30% of siphons. This caused a burrow reduction in 25–75%, respectively, compared to control individuals with intact siphons, in field and laboratory trials. To examine subsequent consequences of reduced burial depth, we exposed nipped and intact clams to potential predators in the laboratory simulating the observed natural clam abundance. Artificially nipped clams were consumed twice as much as control clams. The present results suggest that sympatric croppers contribute to the stock recovery failure by facilitation of lethal predation and that re-seeding to increase the local abundance of M. mactroides should be an essential aspect of conservation efforts in South America.     

石油污染土壤对蚯蚓的致死效应及回避行为的影响

土壤石油污染对生态系统和人类健康有严重影响,其毒害诊断和风险评价的研究日益受到重视。以赤子爱胜蚓(Eiseniafoetida)为受试生物,利用受石油污染的实际土壤,将高浓度污染的土壤与清洁土壤混合,配制成不同污染水平的石油污染土壤,通过直接暴露的方式,分别以蚯蚓急性毒性死亡率和回避反应作为测试终点,评价了石油污染土壤对蚯蚓的生态毒性。结果显示,石油烃污染土壤对蚯蚓的7d-LC50和14d-LC50分别为32.5和29.4g·kg-1,而土壤中石油烃的浓度为8.0g·kg-1时,蚯蚓即有明显回避反应(回避率达80%),前两者为后者的3.7～4.1倍。可见,与急性毒性实验的测试终点LC50相比,行为测试终点对石油烃的响应更灵敏。蚯蚓的回避反应可更加及时地反映出土壤受石油污染的状况,发挥更好的指示作用。     

Hepatic mixed-function oxidases in California flatfishes are increased in contaminated environments and by oil and PCB ingestion

Hepatic mixed-function oxidases (MFOs) were measured in the bothid flatfishes Citharicthys sordidus and C. stigmaeus from relatively uncontaminated and from polluted coastal populations of California, USA, at various times of the year during 1979–1980 and in individuals fed crude oil and polychlorinated biphenyl-augmented food in the laboratory. For C. sordidus, aryl hydrocarbon hydroxylase (AHH) specific activity was generally highest around the Los Angeles County sewage outfall on the Palos Verdes Shelf, intermediate near the 7-mile Hyperion sewage outfall in Santa Monica Bay and around a petroleum seep in the Santa Barbara Channel, and lowest in relatively unpolluted Monterey Bay. For C. stigmaeus, which had about ten times less specific activity than the foregoing species, specimens from the Santa Barbara petroleum seep had significantly greater AHH specific activity than those from Monterey Bay. Fishes from contaminated environments also showed increases of microsomal proteins with molecular weights of 56, 54, 57 and 46×10³; moreover, the content of cytochrome P-450 was elevated in specimens of C. sordidus from such environments. Augmentation of food with seep oil or polychlorinated biphenyls (PCBs) induced significant increases in the specific activity of AHH and amounts of microsomal proteins in C. stigmaeus. Thus, these two species of flatfishes are good candidates for monitoring biologically meaningful levels of petroleum and polychlorinated biphenyls in contaminated environments. Moreover, the mixed function oxidase pattern in fish populations from the Santa Barbara petroleum seep is evidently a functional adaptation to chronic intake of petroleum hydrocarbons.     

Effects of petroleum hydrocarbons on the cytochrome P450 content of the mollusc bivalveMytilus galloprovincialis

The cytochrome P₄₅₀, identified by the characteristic 450 nm spectral peak of its carbon monoxide complex, was detected in the hepatopancreatic microsomes ofMytilus galloprovincialis, collected from Ponteau (Bay of Fos, France) in 1982 and 1983. When mussels were exposed to a variety of petroleum hydrocarbons, the cytochrome P₄₅₀ content increased. It is shown that aromatic hydrocarbons are more effective than alkanes in inducing a high level of cytochrome P₄₅₀. The possible use of this haemoprotein in hydrocarbon pollutant detection is discussed.     

Uptake,metabolism and discharge of polycyclic aromatic hydrocarbons by marine fish

The uptake, metabolism and discharge of two polycyclic aromatic hydrocarbons, ¹⁴C-naphthalene and ³H-3,4-benzopyrene, were studied in 3 species of marine fish (mudsucker or sand goby, Gillichthys mirabilis; sculpin, Oligocottus maculosus; sand dab, Citharichthys stigmaeus). The path of hydrocarbons through the fish included entrance through the gills, metabolism by the liver, transfer of hydrocarbons and their metabolites to the bile, and, finally, excretion. The gall bladder was a major storage site of labeled hydrocarbons and their metabolites. The major product of ²H-3,4-benzypyrene metabolism was tentatively identified as 7,8-dihydro-7,8-dihydroxybenzopyrene. The ¹⁴C-naphthalene was metabolized to 1,2-dihydro-1,2-dihydroxynaphthalene after 24 h exposure. The urine appeared to the major avenne for discharge of labeled hydrocarbon from the body. Our laboratory results indicated that certain polycyelic aromatic hydrocarbons were rapidly taken up from seawater by the above fish, but detoxification mechanisms existed for efficient removal of these compounds from their body tissues.     

Blooms of surf-zone diatoms along the coast of the Olympic Peninsula,Washington. XI. Regeneration of ammonium in the surf environment by the Pacific razor clam Siliqua patula

A 20 month field study was conducted on ammonium excretion rates of the Pacific razor clam Siliqua patula Dixon along the beaches of Washington State, USA. Excretion rates of all those nutrients likely to be regenerated in sufficient quantity to affect surf diatom growth were measured; ammonium appeared to be the most important metabolite. Excretion of ammonium by razor clams far exceeded that by other beach fauna. Ammonium excretion rates of razor clams were positively correlated with shell length, but no correlation between ammonium excretion rate and water temperature was evident. This may be an artifact or may represent some degree of seasonal acclimation of the species to temperature. Weight-specific ammonium excretion rates were negatively related to clam size, indicating a possible large (and unknown) contribution of regenerated ammonium by smaller clams in their first year of growth; smaller clams were rarely captured during this study.Contribution No. 1048 from the Department of Oceanography, University of Washington, Seattle, Washington 98195, USA.     

Fate of petroleum hydrocarbons taken up from food and water by the blue crab Callinectes sapidus

Radiolabeled paraffinic and aromatic hydrocarbons, including benzopyrene, fluorene, naphthalene, methylnaphthalene, methylcholanthrene, hexadecane, heptadecane and dotriacontane, were taken up from food and water by the blue crab Callinectes sapidus. In 2 days, approximately 10% of the benzopyrene and fluorene were taken up from the water when their concentrations were 2.5 and 30 g/l, respectively. When given food with radiolabeled hydrocarbons, 2 to 10% of the hydrocarbons were assimilated by the carbs, with the remainder excreted. After uptake of hydrocarbon from water or food, a major pathway for the elimination of hydrocarbon and metabolites was through fecal material. All hydrocarbons used in the study were metabolized, with similar rates for both paraffinic and aromatic hydrocarbons. More than 50% of the radioactivity assimilated by the crabs was in the hepatopancreas, suggesting that the hepatopancreas was the site of hydrocarbon metabolism. Twenty-five days aftex exposure to radiolabeled hydrocarbons, radioactivity was found only in the hepatopancreas. The hepatopancreas contained highly polar hydrocarbon metabolites, including dihydroxy-compounds and their conjugates, while blood contained both monohydroxy-and dihydroxy-compounds. No evidence was found of storage of hydrocarbons by any of the crab tissues.     

Bioremediation of highly contaminated oilfield soil: Bioaugmentation for enhancing aromatic compounds removal

This study evaluated the effectiveness of different amendments--including a commercial NPK fertilizer, a humic substance （HS）, an organic industrial waste （NovoGro）, and a yeast-bacteria consortium--in the remediation of highly contaminated （up to 6% of total petroleum hydrocarbons） oilfield soils. The concentrations of hydrocarbon, soil toxicity, physicochemical properties of the soil, microbial population numbers, enzyme activities and microbial community structures were examined during the 90-d incubation. The results showed that the greatest degradation of total petroleum hydro- carbons （TPH） was observed with the biostimulation using mixture of NPK, HS and NovoGro, a treatment scheme that enhanced both dehydrogenase and lipase activities in soil. Introduction of exogenous hydrocarbon-degrading bacteria （in addition to biostimulation with NPK, HS and NovoGro） had negligible effect on the removal of TPH, which was likely due to the competition between exogenous and autochthonous microorganisms. None- theless, the addition of exogenous yeast-bacteria consor- tium significantly enhanced the removal of the aromatic fraction of the petroleum hydrocarbons, thus detoxifying the soil. The effect of bioaugmentation on the removal of more recalcitrant petroleum hydrocarbon fraction was likely due to the synergistic effect of bacteria and fungi.     

Trail discrimination signal of Lasius japonicus (Hymenoptera: Formicidae)

Summary. Trail-following behavior of Lasius japonicus was colony-specific in the field, while trail pheromone activity was not. We found that the footprint substance caused colony-specific trail-following behavior only when working in conjunction with the trail pheromone. The footprint substance alone did not lead the workers to follow trails. The substance consisted mainly of hydrocarbons with composition almost identical to that of cuticular hydrocarbons, except for the absence of n-alkanes. Nestmate workers shared footprint hydrocarbon profiles as well as cuticular hydrocarbons, but the profiles differed among colonies. We therefore consider that the footprint hydrocarbon profiles serve as the trail discrimination signal in L. japonicus.     

Biogeochemistry of aromatic hydrocarbons in the benthos of microcosms†

Investigations into the fate of petroleum compounds in the marine environment were carried out using experimental microcosms of two sizes and designs. Aromatic hydrocarbons or No. 2 fuel oil were spiked to the water of a 13 m³ continuous flow system and to a 2281 recycled flow system. The transport and alteration of this oil was traced in the sediment and benthic organisms (Glycera americana, Crepidula sp., and Nephtys incisa) of these microcosms. Measurable contamination was found in both sample types. The aromatic hydrocarbon distribution, including relative isomeric distribution (e.g., C₂‐phenanthrenes) was found to be different in sediment and in organisms from that which was originally introduced to the experimental microcosm. Differences in isomer distribution between Glycera and Crepidula were also detected. Based on the experimental data: molecular weight and specific isomeric form, biochemical processes, solubility, and particle adsorption/desorption influence the fate of petroleum compounds in benthic ecosystems.     

A long-term study of the effects of water-soluble fractions of No. 2 fuel oil on the survival,development rate,and growth of the mud crab Rhithropanopeus harrisii

Mud crabs, Rhithropanopeus harrisii (Gould), were exposed continuously for 6 months after hatching to water-soluble fractions (WSF) of No. 2 fuel oil. Survival, growth and development rate were monitored during this time. The zoeal stages were the most sensitive to fuel oil. A 20% WSF (0.36 ppm total naphthalenes, 1.26 ppm total hydrocarbons) was acutely toxic to these stages. Of the zoeal stages, the first stage appeared to be the most sensitive. The combined duration of the 4 zoeal stages was significantly increased by increasing WSF exposure concentrations. The megalopa and crab stages were not particularly sensitive to continued petroleum hydrocarbon exposure, particularly when compared to zoeal stages. However, mean duration of the megalopa and first crab stages was significantly affected by oil exposure. Individuals which survived the highest exposure concentrations as larvae appeared to grow larger during the crab stages, so that at the end of 6 months comparably staged crabs were equal to or larger than both control crabs and those exposed to low WSF concentrations. Stage distributions at the end of 6 months showed no differences due to WSF exposure. Sex ratios, which could be determined at the end of 6 months, were approximately 1, indicating no sex-related differential sensitivity to WSF exposure, at least as larvae or juveniles. The data indicate that these crabs possess considerable ability to recover from the effects of chronic sublethal exposure to petroleum hydrocarbons. The most deleterious effects of oil pollution on this species may be due to its impact on larval recruitment into the adult population.     

Accumulation of 14C-1-naphthalene by an oceanic and an estuarine copepod during long-term exposure to low-level concentrations

Accumulation of the bi-cyclic aromatic hydrocarbon ¹⁴C-1-naphthalene in adult female Calanus helgolandicus Claus and adult female Eurytemora affinis Poppe in sea water concentrations of hydrocarbon ranging from 0.2 to 992 g/l was studied during exposure periods of up to 15 days as part of an investigation of the possible effects on marine zooplankton of persistent exposure to low levels of petroleum hydrocarbons. With both species the body levels of radioactivity increased rapidly during the first few days of the exposure period, but after exposure for 7 to 8 days to sea water containing 50 g hydrocarbon/l an equilibrium condition was approached; in some experiments where E. affinis was exposed to 1.0 and 10 g hydrocarbon/l for 15 days there was no further increase in body levels of radioactivity after 7 to 8 days. Using a low concentration of hydrocarbon (1 g/l), the quantity of radioactivity accumulated after 10 days was found to be nearly fifty times greater in the smaller species, E. affinis, than in C. helgolandicus, when expressed in terms of body weight. After they had been exposed to the hydrocarbon for several days the copepods contained a considerable proportion of radioactivity that was no longer identifiable as naphthalene and was presumably present as metabolites. Radioactivity accumulated in the copepods after several days was rapidly lost after they were transferred to uncontaminated sea water: e.g. C. helgolandicus lost nearly 90% of its body level of radioactivity in 24 h. Thereafter the rate of loss was greatly reduced, and 5% of the original body level of radioactivity still remained in the copepods at the end of 11 days. Experiments on the breakdown of naphthalene added at low concentrations to sea water samples containing natural microbial populations indicated degradation rates of 0.1 to 0.2 g/l/24 h in oceanic water, and 2.6 g/l/24 h in inshore water samples. The results are discussed in terms of the possible transfer of hydrocarbon to a higher trophic level in areas subjected to constant low-level inputs of petroleum hydrocarbons.     

The influence of fresh weight and water temperature on metabolic rates and the energy budget of Meretrix meretrix Linnaeus

Meretrix meretrix L. was held in the laboratory under simulated natural conditions to measure specific physiological parameters of its energy budget. O₂ consumption rate, NH₃ excretion rate (NR), ingestion rate, faeces excretion rate and scope for growth (SFG) were negatively related in an exponential manner to the fresh weight of the clams at all water temperatures, while almost all metabolic rates of the clams were positively related in an exponential or e-exponential manner with water temperature. However, the co-relationship between metabolic rates and water temperature was not as close as that between metabolic rates and fresh weight of the clam. The combined effect of fresh weight and water temperature was observed on all metabolic rates except for NR and SFG. At all culture temperatures and for all fresh weights of clams used, respiration took the largest percentage of ingested energy (41.5–51.2%), faeces excretion was second (31.0–42.3%), growth third (12.1–15.5%) and urine production last (2.1–5.6%).     

Respiration and nitrogen excretion by the squid Loligo forbesi

The amphipod Corophium volutator (Pallas) of the Bay of Fundy is a major food resource for migrating shorebirds. A relatively low total lipid content (ca 1.7%) was found in summer and fall samples of 1986. Analysis of triacylglycerides which would normally reflect an animal diet through fatty acid input, revealed high proportions of fatty acids characteristic of algae. C. volutator contains hydrocarbons (0.2% of total lipids) with a unimodal distribution in the range C₁₈ to C₃₅ with maximum at C₂₅, and a weak odd-carbon preference (carbon preference index, C.P.I. =1.22). This hydrocarbon pattern shown by C. volutator seems to be biogenic, but due to both a weak odd-carbon preference and a significant content of the isoprenoid phytane, the presence of petroleum hydrocarbons cannot be ruled out. Comparison of the hydrocarbon composition be ruled out. Comparison of the hydrocarbon composition found in C. volutator with hydrocarbons described in the literature for its most likely sources of food suggests that highly degraded material and aged detritus are components of the diet of this deposit feeder, and as important as the local diatom population. No organohalogen pesticides (DDT, dichlorodiphenyl-trichloroethane, or derivatives) or polychlorinated biphenyls (PCBs) were detected in the lipid extracts of C. volutator. Absence of these materials and low hydrocarbon levels explain the continued successful exploitation of this food resource by migrating shorebirds.     

Very long-chain aliphatic hydrocarbons in lipids of mussels (Mytilus edulis) suspended in the water column near petroleum operations off Sable Island,Nova Scotia,Canada

Mussels (Mytilus edulis) suspended in the water column in 1994 and 1995 for the monitoring of oil drilling operations off Sable Island, Nova Scotia were examined for hydrocarbon profiles, particularly aliphatic hydrocarbons. A spring bloom of phytoplankton occurred during the 90-d suspension period in 1995. Hydrocarbons isolated from the 1995 suspended mussels showed very high concentrations of both biogenic hydrocarbons and very long-chain n-alkanes from C₂₀ to C₃₂, initially thought to be petrogenic. Both types of hydrocarbons were either not detected or were only present in trace amounts in the mussels suspended in 1994 at similar sites. The biogenic hydrocarbons in the 1995 mussels were apparently of planktonic origin, from the spring bloom, and were dominated by heneicosahexaene (21:6), followed by pristance, heptadecane, and varions monounsaturated and polyunsaturated phytenes, heptadecenes, nonadecenes and heneicosenes. They could be readily hydrogenated to yield the basic alkanes. The 1995 mussels suspended within 1 km from the oil well platform were probably slightly tainted by petrogenic hydrocarbons, as evidenced by the detection of phytane and high concentrations of total aliphatic hydrocarbons, whereas the mussels suspended 10 km from the platform showed only high concentrations of biogenic hydrocarbons and the novel long-chain n-alkanes. The occurrence of an unusual phytoplankton bloom during the suspension period severely interfered with the petroleum monitoring role of mussels by altering the mussel hydrocrbon profiles through the accumulation into and probably selective depuration of xenobiotic hydrocarbons from the mussel, tissues.J. Parsons (deceased)