Hydrocarbon contamination decreases mating success in a marine planktonic copepod

The mating behavior and the mating success of copepods rely on chemoreception to locate and track a sexual partner. However, the potential impact of the water-soluble fraction of hydrocarbons on these aspects of copepod reproduction has never been tested despite the widely acknowledged acute chemosensory abilities of copepods. I examined whether three concentrations of the water-soluble fraction of diesel oil (0.01%, 0.1% and 1%) impacts (i) the swimming behavior of both adult males and females of the widespread calanoid copepod Temora longcornis, and (ii) the ability of males to locate, track and mate with females. The three concentrations of the water-soluble fraction of diesel oil (WSF) significantly and non-significantly affect female and male swimming velocities, respectively. In contrast, both the complexity of male and female swimming paths significantly decreased with increasing WSF concentrations, hence suggesting a sex-specific sensitivity to WSF contaminated seawater. In addition, the three WSF concentrations impacted both T. longicornis mating behavior and mating success. Specifically, the ability of males to detect female pheromone trails, to accurately follow trails and to successfully track a female significantly decreased with increasing WSF concentrations. This led to a significant decrease in contact and capture rates from control to WSF contaminated seawater. These results indicate that hydrocarbon contamination of seawater decreases the ability of male copepods to detect and track a female, hence suggest an overall impact on population fitness and dynamics.     

Advance in the toxic effects of petroleum water accommodated fraction on marine plankton

Recently, the impact of petroleum pollution on marine plankton has been complemented by a great concern. This review summarizes the reports about toxic effects of oil water accommodated fraction (WAF) on marine phytoplankton, zooplankton and early life stage of animal. For the oil WAF, toxicants are mainly composed of the aromatic hydrocarbons, such as the benzene hydrocarbons and polycyclic aromatic hydrocarbons (PAHs) with 2–5 rings. The oil WAF, especially the PAHs, can be accumulated in plankton due to their great lipophilic abilities, and thus elicites various deleterious effects. Toxicological tests show that marine plankton is very sensitive to the petroleum WAF, as the order of median effective/lethal concentration is merely μg/L or mg/L. There are species and developmental stages differences of plankton tolerance to petroleum WAF, and the toxicity of different oil WAF is various. Generally, its toxicity enhances with increasing carbonic chain length and benzene ring number. Many studies on the acute and sub-acute toxic effects of oil WAF have been done, however few researches on its chronic toxic effects has been carried out till now. Besides, most reports focused on the levels from molecule to individual, though very little work of petroleum toxic effects has ever been performed on the marine plankton population or community levels. Therefore, it is necessary to continue these studies in future.     

Petroleum biodegradation in marine environments

Petroleum-based products are the major source of energy for industry and daily life. Petroleum is also the raw material for many chemical products such as plastics, paints, and cosmetics. The transport of petroleum across the world is frequent, and the amounts of petroleum stocks in developed countries are enormous. Consequently, the potential for oil spills is significant, and research on the fate of petroleum in a marine environment is important to evaluate the environmental threat of oil spills, and to develop biotechnology to cope with them. Crude oil is constituted from thousands of components which are separated into saturates, aromatics, resins and asphaltenes. Upon discharge into the sea, crude oil is subjected to weathering, the process caused by the combined effects of physical, chemical and biological modification. Saturates, especially those of smaller molecular weight, are readily biodegraded in marine environments. Aromatics with one, two or three aromatic rings are also efficiently biodegraded; however, those with four or more aromatic ring are quite resistant to biodegradation. The asphaltene and resin fractions contain higher molecular weight compounds whose chemical structures have not yet been resolved. The biodegradability of these compounds is not yet known. It is known that the concentrations of available nitrogen and phosphorus in seawater limit the growth and activities of hydrocarbon-degrading microorganisms in a marine environment. In other words, the addition of nitrogen and phosphorus fertilizers to an oil-contaminated marine environment can stimulate the biodegradation of spilled oil. This notion was confirmed in the large-scale operation for bioremediation after the oil spill from the Exxon Valdez in Alaska. Many microorganisms capable of degrading petroleum components have been isolated. However, few of them seem to be important for petroleum biodegradation in natural environments. One group of bacteria belonging to the genus Alcanivorax does become predominant in an oil-contaminated marine environment, especially when nitrogen and phosphorus fertilizers are added to stimulate the growth of endogenous microorganisms.     

Effect of crude oil petroleum hydrocarbons on protein expression of the prawn Macrobrachium borellii

Hydrocarbon pollution is a major environmental threat to ecosystems in marine and freshwater environments, but its toxicological effect on aquatic organisms remains little studied. A proteomic approach was used to analyze the effect of a freshwater oil spill on the prawn Macrobrachium borellii. To this aim, proteins were extracted from midgut gland (hepatopancreas) of male and female prawns exposed 7 days to a sublethal concentration (0.6 ppm) of water-soluble fraction of crude oil (WSF). Exposure to WSF induced responses at the protein expression level. Two-dimensional gel electrophoresis (2-DE) revealed 10 protein spots that were differentially expressed by WSF exposure. Seven proteins were identified using MS/MS and de novo sequencing. Nm23 oncoprotein, arginine methyltransferase, fatty aldehyde dehydrogenase and glutathione S-transferase were down-regulated, whereas two glyceraldehyde-3-phosphate dehydrogenase isoforms and a lipocalin-like crustacyanin (CTC) were up-regulated after WSF exposure. CTC mRNA levels were further analyzed by quantitative real-time PCR showing an increased expression after WSF exposure. The proteins identified are involved in carbohydrate and amino acid metabolism, detoxification, transport of hydrophobic molecules and cellular homeostasis among others. These results provide evidence for better understanding the toxic mechanisms of hydrocarbons. Moreover, some of these differentially expressed proteins would be employed as potential novel biomarkers.     

Unexpected Interaction with Dispersed Crude Oil Droplets Drives Severe Toxicity in Atlantic Haddock Embryos

The toxicity resulting from exposure to oil droplets in marine fish embryos and larvae is still subject for debate. The most detailed studies have investigated the effects of water-dissolved components of crude oil in water accommodated fractions (WAFs) that lack bulk oil droplets. Although exposure to dissolved petroleum compounds alone is sufficient to cause the characteristic developmental toxicity of crude oil, few studies have addressed whether physical interaction with oil micro-droplets are a relevant exposure pathway for open water marine speices. Here we used controlled delivery of mechanically dispersed crude oil to expose pelagic embryos and larvae of a marine teleost, the Atlantic haddock (Melanogrammus aeglefinus). Haddock embryos were exposed continuously to two different concentrations of dispersed crude oil, high and low, or in pulses. By 24 hours of exposure, micro-droplets of oil were observed adhering and accumulating on the chorion, accompanied by highly elevated levels of cyp1a, a biomarker for exposure to aromatic hydrocarbons. Embryos from all treatment groups showed abnormalities representative of crude oil cardiotoxicity at hatch (5 days of exposure), such as pericardial and yolk sac edema. Compared to other species, the frequency and severity of toxic effects was higher than expected for the waterborne PAH concentrations (e.g., 100% of larvae had edema at the low treatment). These findings suggest an enhanced tissue uptake of PAHs and/or other petroleum compounds from attached oil droplets. These studies highlight a novel property of haddock embryos that leads to greater than expected impact from dispersed crude oil. Given the very limited number of marine species tested in similar exposures, the likelihood of other species with similar properties could be high. This unanticipated result therefore has implications for assessing the ecological impacts of oil spills and the use of methods for dispersing oil in the open sea.     

Toxic effect of water-soluble fractions of crude, refined, and weathered oils on the growth of a marine bacterium.

The water-soluble fractions of three crude and two refined oils reduced the growth rate and maximum cell density of the marine bacterium Serratia marinorubra grown in batch culture. The weathering of a crude and a refined oil was simulated in the laboratory. The water-soluble fractions remaining from this process were more toxic to S. marinorubra than were the parent unweathered oils. Increases in the magnitude of toxic effect of 3 to 30 times were observed as a function of decreasing the concentration of yeast extract in the cultures from 0.1 to 0.05 and 0.01%. The toxicity did not correlate with the concentration of total water-soluble fraction or of aromatic hydrocarbons in the water-soluble fraction. Affected cultures did not exhibit a residual toxicity after being back-inoculated into control media.     

Acute toxicity of the water-soluble fraction of spent lubricating oil on the African catfish Clarias gariepinus

Static tests were employed to assess the acute toxicity of the water-soluble fraction (WSF) of spent automotive lubricating oil (of mixed SAE grades) on Clarias gariepinus, a freshwater fish commonly cultured in Nigeria. Median lethal concentrations (LC50) of the WSF were found to decrease as a function of exposure time from 690+/-21 (after 24 h) to 513+/-58 mg/l (after 96 h). The characteristics of the WSF such as mean acidity (pH 6.6), turbidity (40 NTU), total dissolved solids (TDS; 40 mg/l) and significantly reduced (P<0.05) dissolved-oxygen (DO) values (1.44 mg/l) were not compliant with existing standards set for discharged effluents. The solubility of the detected straight-chain aliphatics ranked as C14>C16>C32>C18>C28; that of the simple aromatics was ortho-xylene>para-xylene; and that of the polyaromatic hydrocarbons (PAHs) was acenaphthylene>9H-fluorene>naphthalene>anthracene>phenanthrene>chrysene>benzo[k]fluoranthene>benzo[a]pyrene>benzo[b]fluoranthene, most of which being serious carcinogens. These oil constituents and the overall physico-chemical properties of the WSF are expected to act synergistically on the test organism (C. gariepinus), eliciting the quantal responses observed. The toxicity of the WSF points to the base constituent, oxidative degradation, and mechano-chemical reactions associated with aged crankcase oils. These oils, therefore, should definitely no longer be disposed into water streams or landscape, not even at sub-lethal concentrations, because of the inherent toxicity of their soluble fractions and the associated danger of bioaccumulation.     

Inhibition of light-induced pH increase and O2 evolution of marine microalgae by water-soluble components of crude and refined oils.

Light-induced alkalinization of the extracellular medium was found to be a common feature of the primary photosynthetic process of several marine microalgae. The light-induce PH increase of suspensions of whole cells was immediately and severely inhibited by a single dose of water-soluble components from crude and fuel oils. Differential effects on the rates of microalgal photosynthetic O2 evolution and cell suspension pH increase suggest different toxicity mechanisms of the water-soluble components of no. 2 fuel oil as compared with Southern Louisiana and Jay Crude oils. These short-term studies on the nature of sublethal petroleum toxicity to microalgae indicate that the primary effect may be through direct action on the energy-yielding electron transport systems.     

Inhibition of light-induced pH increase and O2 evolution of marine microalgae by water-soluble components of crude and refined oils.

Light-induced alkalinization of the extracellular medium was found to be a common feature of the primary photosynthetic process of several marine microalgae. The light-induce PH increase of suspensions of whole cells was immediately and severely inhibited by a single dose of water-soluble components from crude and fuel oils. Differential effects on the rates of microalgal photosynthetic O2 evolution and cell suspension pH increase suggest different toxicity mechanisms of the water-soluble components of no. 2 fuel oil as compared with Southern Louisiana and Jay Crude oils. These short-term studies on the nature of sublethal petroleum toxicity to microalgae indicate that the primary effect may be through direct action on the energy-yielding electron transport systems.     

Effect of water-soluble fraction of Cook Inlet crude oil on swimming performance and plasma cortisol in juvenile coho salmon (Oncorhynchus kisutch)

Swimming performance of juvenile coho salmon decreased and plasma cortisol increased, following 48-hr exposure to the water-soluble fraction (WSF) of Cook Inlet crude oil at 75% of the LC50. Exposure to 25 and 50% of the LC50 did not significantly reduce swimming performance. Plasma cortisol concentrations were highest in fish exposed to both the combined stress of WSF exposure and of forced swimming in a stamina tunnel.     

Corticosteroid and thyroid responses of larval and juvenile turbot exposed to the water-soluble fraction of crude oil

Turbot larvae (24–590 degree C days; 2–32 days post-hatch) and juveniles (1345 degree C days; 98 days post-hatch), were exposed for 6 h to 25, 33 and 50% water-soluble fraction (WSF) of crude oil in either static or flow-through test systems. Larvae showed generalized primary endocrine responses, identified by elevated whole body cortisol content from as early as 2 days post-hatch. In older larvae and juveniles, the response was related to the WSF concentration. This dose-response relationship was not apparent in younger and yolk-sac larvae. Whole body thyroxine content of turbot larvae exposed to the WSF of crude oil was increased, but triiodothyronine content remained stable. Aromatic hydrocarbon concentrations [benzene, toluene, ethylbenzene and xylene (BTEX) and naphthalenes] remained constant during flow-through tests, but 65% of the initial level of BTEX hydrocarbons and 40% of the naphthalenes were lost during static exposures. Larval mortalities increased with exposure to an increasing concentration of crude oil WSF. Larval activity was significantly reduced even at the lowest WSF concentration.     

Intranuclear crystalloids of Antarctic sea urchins as a biomarker for oil contamination

Because of human actions, biomarkers have become important to detect and mitigate pollution. This study showed that crystalloids can be a biomarker for analyses of low levels of water-soluble fractions of oil (WSF). Antarctic sea urchins (Sterechinus neumayeri) from regions free of pollution were exposed for 2, 5, 10 and 15 days at different levels of WSF (0.4, 0.8 and 1.2 ppm). No significant differences were observed in the phagocytic rates or the germicide capacity for the yeast Saccharomyces cerevisiae; however, there was a significant increase in the quantity of intranuclear iron crystalloids in phagocytic amoebocytes of urchins exposed to higher levels of WSF. This study characterizes histological alterations in crystalloids of S. neumayeri that could be used as a biomarker for oil contaminants, with a simple and inexpensive protocol.     

Influence of the water-soluble fraction of petroleum on photosynthesis and chemical defenses in two sympatric seaweeds

In this work, we aimed to evaluate the effect of the water-soluble fraction (WSF) of petroleum on the photosynthesis and chemical defenses of two sympatric members of the brown algal family Dictyotaceae, Dictyota caribaea and Stypopodium zonale. The algae were cultivated in incubator chambers for 8 days in flasks containing the WSF or seawater as a control laboratory cultivation. Measurements of chlorophyll fluorescence revealed distinct responses between the two species. Stypopodium zonale was less tolerant than D. caribaea as shown by a more intense diminishing of the potential quantum yield of photosystem II (F _v/F _m) as well as by the reduction of the intrinsic photosynthetic capacity observed in parameters obtained from the rapid light curves. Feeding experiments revealed that the WSF reduced strongly the palatability of the lipophilic extracts from both algae to the herbivore crab Pachygrapsus transversus (Gibbes). The analyses of the chemical profiles from these extracts by chromatographic techniques showed that the effect of the WSF on secondary metabolites production could only be observed in D. caribaea. Our results indicated that these sympatric species showed distinct tolerances to the WSF where S. zonale was more sensitive than D. caribaea. On the other hand, both species responded to the WSF with a more deterrent behavior to herbivory. Possible ecological consequences are discussed.     

石油污染对海洋浮游植物群落生长的影响

为了解石油污染对海洋浮游植物的慢性毒性影响,运用实验生态学手段,在不同季节对采自浙江省乐清湾的浮游植物进行原油污染慢性(15 d)毒性效应研究。通过对浮游植物叶绿素a(chla)和细胞密度的测定,发现高浓度(≥2.28 mg/L)原油胁迫对浮游植物的生长有极显著抑制作用(P＜0.001),表现为浮游植物chla和细胞密度下降;低浓度(≤1.21 mg/L)不会抑制浮游植物生长,反而可促进其生长。各个季节原油对浮游植物的慢性毒性效应存在差异,表现为不同自然水温下浮游植物对原油胁迫的耐受性不同,夏季浮游植物对油污染的耐受性较冬季差。浮游植物chla和细胞密度有极显著的正相关性(P＜0.001)。石油污染对浮游植物的影响随培养时间延长存在差异,培养后期石油烃逐渐在水中降解,低浓度试验组中浮游植物可恢复生长。     

Individual to population level effects of South Louisiana crude oil water accommodated hydrocarbon fraction (WAF) on a marine meiobenthic copepod

Acute toxicities of crude oil and crude oil water accommodated hydrocarbon fraction (WAF) are relatively well documented, but data on the biological effects of chronic exposures to WAF on species and populations are scarce. South Louisiana Sweet crude oil was used to assess the effects of crude oil WAF on the copepod Amphiascus tenuiremis' survival, development and reproduction. Effects were evaluated using a 96-well microplate full life-cycle toxicity test, a test that allows tracking of individuals from the nauplius stage to sexual maturation and reproduction. Briefly, 24-h hatched nauplii were followed to adulthood (n_i = ≥ 120 nauplii/treatment) in individual glass-coated microplate wells containing 200 μL of seawater solution. Treatments consisted of 10%, 30%, 50% and 100% Louisiana WAF, with seawater used as control. Nauplii were monitored through development to adulthood, and sexually mature virgin copepods were mated pairwise in wells containing original rearing treatments. Nauplius-to-copepodite survival was reduced by 57% in exposures to 100% WAF, relative to controls (88 ± 3%), and copepodite-to-adult survival was reduced by 18% in the 50% WAF, relative to controls (98 ± 3%). Analysis of development curves showed that nauplii in the 10% WAF developed significantly faster into copepodites, while nauplii in the 50% WAF developed significantly slower than controls. Although the naupliar developmental rate in the 100% WAF was not significantly different from the control, these nauplii showed an average 1.4 day delay in development into copepodites. Similarly, copepodite development into mature females and males was significantly enhanced by 1.2 to 1.8 days and delayed by 1.9 to 2.2 days (p < 0.05) in the 10% and 50% WAFs, respectively, compared to controls. Although the copepodite developmental rate in the 100% WAF was not significantly different from the control, these copepodites still showed an average 1.5 and 2.1 day delay in development into females and males, respectively. Analysis of reproductive endpoints showed that fertility was the only endpoint negatively affected by WAFs; reproductive failure increased by 30% and 41% in exposures to 30% and 100% WAF, respectively, compared to controls (3.33 ± 4.71%). Leslie matrix population projections based on empirical microplate data indicated lower production rates through three generations of exposure to WAFs. Furthermore, a comparison between NIST and Louisiana crude oil WAFs using the same life-cycle approach indicated a greater chronic toxicity for the Louisiana WAF and an overall developmental delay in exposures to high WAFs (50% and 100% WAFs) from both crude oil types.     

石油烃污染对海洋浮游植物群落的短期毒性效应

海上溢油和船舶含油废水排放等石油污染给海洋生态环境带来严峻考验。为了解石油污染对海洋浮游植物的短期群落效应, 于实验室对采自浙江省乐清湾海域的四季浮游植物进行了14天的微宇宙实验研究。通过测定不同浓度原油水溶性成分(water accommodated fraction, WAF)胁迫下浮游植物群落各种类的细胞密度, 发现原油WAF对浮游植物群落的种类数、多样性、均匀度和优势种组成及优势度均有极显著影响。原油污染组的种类数均低于对照组, 多样性指数也总体上低于对照组, 但均匀度指数无明显规律。WAF对各季节浮游植物群落优势种的影响存在季节、浓度梯度和种类差异: 1)在高浓度WAF (≥2.28 mg·L^-1)胁迫下, 各季节浮游植物群落中中肋骨条藻(Skeletonema costatum)的优势度均呈升高趋势, 除秋季外, 长菱形藻(Nitzschia longissima)在其余季节的优势度均显著下降, 另外秋季实验中微小原甲藻(Prorocentrum minimum)的优势度呈先升高后下降的趋势, 秋冬季斜纹藻(Pleurosigma sp.)和念珠直链藻(Melosira moniliformis)的优势度呈下降趋势; 2)在低浓度WAF (≤1.16 mg·L^-1)胁迫下, 除秋季中肋骨条藻仍为绝对优势种外, 其余季节该种微藻的优势度均呈下降趋势, 春、夏季长菱形藻的优势度逐渐升高, 而冬季则是念珠直链藻的优势度不断增加。原油WAF影响下, 可能r策略种将逐渐取代k策略种, 导致浮游植物群落趋于小型化。     

Comparative effects of the water accommodated fraction of three oils on mussels—3. Quantitative histochemistry of enzymes related to the detoxication metabolism

1. The reaction products derived from the histochemical demonstration of catalase and gamma-glutarnyl transpeptidase (GGT) activities have been quantified in digestive tissue cells of mussels treated for 21, 49 and 91 days with the water accommodated fraction (WAF) of two different crude oils and of one commercial lubricant oil.2. Although not in a dose-dependent manner, the mean absorbance of the catalase reaction product is significantly increased (up to 40% increases over control values) in digestive tubule cells after exposure to the 3 types of WAF.3. These results could support the hypothesis that petroleum hydrocarbon toxicity is mediated in part by an increased generation of oxygen free radicals.4. The elevation of catalase activity in digestive tubule cells could then represent an adaptative response that exerts a protective role against petroleum hydrocarbon-induced free radical toxicity.5. Mean absorbance values of GGT, enzyme involved in phase II metabolism of electrophilic xenobiotics that catalyzes the second step of the mercapturic acid pathway, is transiently decreased under certain hydrocarbon exposure-conditions in both digestive tubule cells and stomach cells.6. This reduction in GGT activity is discussed in relation with the overall effects of organic xenobiotics on mussel biotransformation enzymes.     

Influence of some environmental variables on the ascorbic acid status of striped mullet, Mugil cephalus Linn., tissues. III. Effects of exposure to oil

Exposure to water-soluble fractions (WSF) of a crude and two fuel oils altered the ascorbic acid (AsA) content of several striped mullet, Mugil cephalus , tissues. Exposure to sublethal concentrations of all three WSFs caused a depletion of AsA reserves in brain, gill, kidney and liver tissues, but not in muscle. There was a marked decline in AsA stores in kidney and gill tissues after only one day of exposure to WSFs of both crude and fuel oils. Liver AsA concentrations were significantly depleted after one week of oil exposure. Brain AsA content was only significantly depleted during chronic exposure to the highest oil concentration (20% WSF). A dose-dependent depletion of AsA reserves in the liver but not in the other tissues was observed one week after a single exposure to 2–20% WSFs of a No. 2 fuel oil. Exposure to 20% WSF of the No. 2 fuel oil caused a 47% decrease in liver AsA content one week later. Hepatic concentrations were still significantly depleted after 15 days, but had returned to control levels 20 days after the initial exposure. The data suggest that the depletion of tissue AsA reserves in fish inhabiting oil-contaminated environments could be sufficient on occasions to lead to AsA deficiency.     

Surfactants in microbiology and biotechnology: Part 2. Application aspects

Surfactants are amphiphilic compounds which can reduce surface and interfacial tensions by accumulating at the interface of immiscible fluids and increase the solubility, mobility, bioavailability and subsequent biodegradation of hydrophobic or insoluble organic compounds. Chemically synthesized surfactants are commonly used in the petroleum, food and pharmaceutical industries as emulsifiers and wetting agents. Biosurfactants produced by some microorganisms are becoming important biotechnology products for industrial and medical applications due to their specific modes of action, low toxicity, relative ease of preparation and widespread applicability. They can be used as emulsifiers, de-emulsifiers, wetting and foaming agents, functional food ingredients and as detergents in petroleum, petrochemicals, environmental management, agrochemicals, foods and beverages, cosmetics and pharmaceuticals, and in the mining and metallurgical industries. Addition of a surfactant of chemical or biological origin accelerates or sometimes inhibits the bioremediation of pollutants. Surfactants also play an important role in enhanced oil recovery by increasing the apparent solubility of petroleum components and effectively reducing the interfacial tensions of oil and water in situ. However, the effects of surfactants on bioremediation cannot be predicted in the absence of empirical evidence because surfactants sometimes stimulate bioremediation and sometimes inhibit it. For medical applications, biosurfactants are useful as antimicrobial agents and immunomodulatory molecules. Beneficial applications of chemical surfactants and biosurfactants in various industries are discussed in this review.