Concentrations Of Water Accommodated Fraction Research Articles

Differential effects of petroleum hydrocarbons on the growing development and physiological characteristics of Ulva species.

To compare the different effects of petroleum hydrocarbons on intertidal Ulva macroalgae, three dominant Ulva species (U. prolifera, U. linza, and U. lactuca) were exposed to two water-accommodated fractions (WAFs) of 0# diesel oil and crude oil at three concentration levels. The results indicated that two WAFs had significant concentration effects on the physiological characteristics of Ulva, the toxicity of 0# diesel oil was greater than crude oil, and crude oil had hormesis effect. Exposure of high WAFs concentrations, the growth, pigment, carbohydrate, and protein contents of Ulva were inhibited, while the antioxidant system was activated. In addition, the integrated biomarker response (IBR) indicated that U. prolifera had higher resistance to WAFs than U. linza and U. lactuca. Considering that U. prolifera is the main species of green tide in the Yellow Sea (YS) of China, the comparative effects of WAFs on different development stages of U. prolifera were also explored. The results showed that spore was the most sensitive to WAFs, while adult thalli was the most tolerant. The increased resistance of U. prolifera thalli and the hormesis effect triggered by crude oil may influence the outbreak scale of green tides. This study provides a new perspective for understanding the formation of green tides in the YS.

Water-accommodated fractions of crude oil and its mixture with chemical dispersant impairs oxidase stress and energy metabolism disorders in Oryzias melastigma embryos

In this study, marine medaka (Oryzias melastigma) embryos were exposed to different concentrations of water-accommodated fractions (WAFs) and chemically enhanced water-accommodated fractions (CEWAFs) of Oman crude oil for 14 d by semi-static exposure methods. The effects on growth and development and energy metabolism process were evaluated. Results showed that embryo survival and hatchability were decreased in a dose-dependent manner with an increase in the concentration of petroleum hydrocarbon compounds, whereas the malformation exhibited a dose-dependent increase. Compared to the control, the adenosine triphosphate (ATP) content and Na+-K+–ATPase (NKA) activities of embryos exposed to both WAFs and CEWAFs were reduced, while intracellular reactive oxygen species (ROS) levels and NADH oxidase (NOX) activities were increased. Our study demonstrated that exposure to crude oil dispersed by chemical dispersant affected the growth and development of marine medaka embryos, caused oxidative stress while produced a series of malformations in the body and dysregulation in energy metabolism. In comparison, the toxic effects of chemically dispersed crude oil might be more severe than the oil itself in the equivalent diluted concentration treatment solution. These would provide more valuable and reliable reference data for the use of chemical dispersants in oil spills.

Acute inhibition of bacterial growth in coastal seawater amended with crude oils with varied photoreactivities

The increased potential for contamination of seawater by crude oils requires studies of bacterial biodegradation potential, but little is known of the differential negative impacts of oils on bacterial growth. No two wells generate chemically identical oils; and importantly, solar exposure of crude oil may differentially affect the bacterial response. Elucidating the role that sunlight plays on the potential toxicity of spilled crude oils is imperative to understanding how oil spills might affect microbes in the tropical and subtropical waters of Florida. This study examined light exposure of six different crude oils, and subsequent microbial responses to altered oils. Marine bacterioplankton heterotrophic activities were measured via3H-leucine incorporation after the addition of oils’ water accommodated fractions (WAFs) that were created under varied solar conditions. Inhibition of production increased with higher concentrations of WAFs, but dose-response trends varied among the oils. Increased solar exposure during WAF preparation generally led to more inhibition, but trends varied among oils. WAFs were also prepared under different parts of the solar spectrum. Solar-irradiated WAFs resulted in significant but variable acute toxicity vs. dark counterparts. Solar-induced toxicity was primarily a result of visible and not ultraviolet light exposure. Results indicate responses to oil spills are highly dependent on the source of the oil and solar conditions at the time and location of the spill. The data presented here demonstrate the importance of photochemical changes and oil source in modulating microbial activity and bioremediation potential.

Early developmental toxicity of Atlantic salmon exposed to conventional and unconventional oils

Atlantic salmon is an important species for Canadian culture and economy and its importance extends beyond Canada to Scandinavia and Western Europe. However, it is a vulnerable species facing decline due to habitat contamination and destruction. Existing and new Canadian pipeline projects pose a threat to salmonid habitat. The effects of diluted bitumen (dilbit), the main oil circulating in pipelines, are less studied than those of conventional oils, especially during the critical early embryonic developmental stage occurring in freshwater ecosystems. Therefore, this study aimed to compare the effects of water-accommodated fractions (WAF) of the Clearwater McMurray dilbit and the Lloydminster Heavy conventional oil on Atlantic salmon embryos exposed either from fertilization or from eyed stage. The dilbit contained the highest concentrations of low molecular weight (LMW) compounds (including BTEX and C6-C10), while the conventional oil contained the highest concentrations of PAHs. The Clearwater dilbit caused a higher percentage of mortality and malformations than the conventional oil at similar WAF concentrations. In addition, the embryos exposed from fertilization suffered a higher mortality rate, more developmental delays, and malformations than embryos exposed from the eyed stage, suggesting that early development is the most sensitive developmental stage to oil exposure. Gene expression and enzymatic activity of the detoxification phase I and II enzymes (CYP1A and GST) were measured. Data showed increases in both cyp1a expression and GST activity with increasing WAF concentrations, while gst expression was not affected by the exposures. Also, gene expression of proteins involved in the biotransformation of vitamin A and DNA damage repair were modified by the oil exposures. Overall, this study indicates that Atlantic salmon is mostly affected by oil exposure at the beginning of its development, during which embryos accumulate deformities that may impact their survival at later life stages.

Comparative toxicity of conventional and unconventional oils during rainbow trout (Oncorhynchus mykiss) embryonic development: From molecular to health consequences

Canadian freshwater ecosystems are vulnerable to oil spills from pipelines, which contain mostly diluted bitumen. This study aimed to compare the toxicity of a dilbit and a conventional oil on developing rainbow trout. A total of five exposure scenarios were performed, from 10 to 43 days, using water-accommodated fraction (WAF) with an initial loading of 1:9 oil to water ratio (w/v) in a range of dilutions from 0.32 to 32% WAF, respectively, with TPAH and VOC concentrations from 2.41 to 17.5 μg/L and 7.94–660.99 μg/L, and with or without a recovery period. Following the five exposures, several endpoints were examined, including survivorship, morphometrics, gene expression, and enzymatic activity. Significant mortality rates were measured for the highest WAF concentration of the dilbit in all five exposures (60–100% mortality at 32% WAF). In comparison, the highest WAF concentration of the conventional oil induced significant mortality in three out of the five exposure (from 35 to 100% mortality at 32% WAF). Hatching delays were noted in embryos exposed to both oils. Developmental delays were observed in dilbit-exposed embryos and are suspected to be an indicator of reduced survivorship after hatching. The induced expression of cyp1a remained a reliable biomarker of exposure and of fish malformations, though it did not always predict mortality. Using CYP1A activity in combination with cyp1a may bring more insights in studies of oil risk assessment. This study demonstrates that dilbits are more toxic to early life stages compared to conventional oils and highlights the need to consider the most sensitive stage of development when performing risk assessment studies on oils.

Influence of fuel oil on Platymonas helgolandica: An acute toxicity evaluation to amino acids.

It is highly likely that the toxicity of water accommodated fractions (WAF) will influence marine microalgae, and consequently lead to potential risk for the marine ecological environment. However, it was often neglected whether WAF can influence the transformation of relative compounds in organisms. The metabolism of amino acids (AAs) can be used to track physiological changes in microalgae because amino acids are the basis of proteins and enzymes. In this study, using marine Chlorophyta Platymonas helgolandica as the test organism, the effects of different concentrations of WAF on AA compositions and stable carbon isotope ratios (δ13C) of individual AAs of Platymonas helgolandica were investigated. The results showed that the WAF of #180 fuel oil had an obvious suppressing effect on the growth and chlorophyll a content of microalgae. The growth inhibitory rate at 96h was 80.66% at a WAF concentration of 0.50mgL-1 compared with the control. Furthermore, seven among the 16 AAs, including alanine, cysteine, proline, aspartic acid, lysine, histidine and tyrosine, had relatively high abundance. Under the glycolysis pathway, the cysteine abundance was higher than control, meaning that the biosynthesized pathway of alanine through cysteine as a precursor could be damaged. Phosphoenolpyruvate (PEP) was an important synthesis precursor of alanine (leucine) and aromatic AA family (Phenylalanine and tyrosine), and played an important role in δ13CAAs fractionation under the WAF stress. Under the TCA pathway, to protect cell metabolism activities under WAF stress, the δ13C value of threonine and proline abundance in microalgae with the increase in WAF stress. Therefore, δ13CAAs fractionation can be used as a novel method for toxicity evaluation of WAF on future.

Effects of chronic exposure to water accommodated fraction (WAF) of light crude oil on gut microbiota composition of the lined sole (Achirus lineatus)

Exposure of marine fish to hydrocarbon compounds from crude oil can cause physiological and ecological alterations that can result in several cytotoxic, genotoxic, and teratogenic damages. One consequence of this exposure is the dysbiosis of the gut microbiota, where the normal bacterial composition is modified. Herein, we assessed the effect of the exposure to water accommodated fraction (WAF) of a light crude oil into the gut microbiota of a native species, the lined sole Achirus lineatus, a benthonic fish widely distributed in the Gulf of Mexico (GoM). We performed a chronic bioassay using two WAF concentrations (5 and 10% v/v), collecting lined sole entire gastrointestinal tracts for microbiota analyses at two timepoints, 14 and 28 days. Changes in the gut microbiota composition were determined by high throughput amplicon sequencing of the gene 16S rRNA. Diversity analyses showed that WAF exposure produced similar changes in the microbiota composition at both concentrations. Metagenomic functional prediction showed that these alterations could result in a shift in the gut redox status, towards a more anoxygenic environment. Enrichment of bacteria capable of use hydrocarbon as carbon source seems to be fast regardless time of exposure or WAF concentrations. Our results suggest that chronic WAF exposure can cause a dysbiosis in this benthic native species from the GoM.

Effect of water accommodated fractions of fuel oil on fixed carbon and nitrogen by microalgae: Implication by stable isotope analysis.

Effect of water accommodated fractions (WAF) of #180 fuel oil on fixed carbon and nitrogen in microalgae was studied by stable isotopes. Platymonas helgolandica, Heterosigma akashiwo and Nitzschia closterium were exposed to five WAF concentrations for 96h. The δ13C value of microalgae was significantly lower than that of the control group, indicated that carbon was limited in the WAF concentrations. The δ13C value of microalgae appeared peak valley at 48h in control group, corresponding to the enhanced capacity in carbon fixation during microalgae photosynthesis. The physiological acclimation capacity of microalgae was revealed by the occurrence time when the δ13C value was in peak valley, and thus the physiological acclimation capacity of microalgae decreased in the order of Nitzschia closterium>Heterosigma akashiwo>Platymonas helgolandica. Principal component analysis (PCA) were applied to the δ13C value in order to verify the "hormesis" phenomenon in microalgae. The δ13C value could discriminate between stimulatory effects at low doses and inhibitory effects at high doses. In addition, the present study also investigated the effect of the nitrogen on microalgae growth. Because microalgae could still absorb the NO3-N and release of NO2-N and NH4-N in present study, the nitrogen cycle in microalgae was in the equilibrium status. The δ15N value in microalgae exhibited no obvious change with the increasing of WAF concentrations at the same time. However, due to the enrichment of nitrogen, the δ15N value first increased gradually with the time and finally was stable. Overall, the fractionation of carbon and nitrogen stable isotopes illustrated that the effect of carbon on the growth of microalgae was more prominent than nitrogen. Stable isotopes was used to investigate the influence of WAF on fixed carbon and nitrogen in microalgae growth, providing a fundamental theoretical guidance for risk assessment of marine ecological environment.

Acute oil exposure reduces physiological process rates in Arctic phyto- and zooplankton.

Arctic shipping and oil exploration are expected to increase, as sea ice extent is reduced. This enhances the risk for accidental oil spills throughout the Arctic, which emphasises the need to quantify potential consequences to the marine ecosystem and to evaluate risk and choose appropriate remediation methods. This study investigated the sensitivity of Arctic marine plankton to the water accommodated fraction (WAF) of heavy fuel oil. Arctic marine phytoplankton and copepods (Calanus finmarchicus) were exposed to three WAF concentrations corresponding to total hydrocarbon contents of 0.07 mg l-1, 0.28 mg l-1 and 0.55 mg l-1. Additionally, the potential phototoxic effects of exposing the WAF to sunlight, including the UV spectrum, were tested. The study determined sub-lethal effects of WAF exposure on rates of key ecosystem processes: primary production of phytoplankton and grazing (faecal pellet production) of copepods. Both phytoplankton and copepods responded negatively to WAF exposure. Biomass specific primary production was reduced by 6, 52 and 73% and faecal pellet production by 18, 51 and 86% with increasing WAF concentrations compared to controls. The phototoxic effect reduced primary production in the two highest WAF concentration treatments by 71 and 91%, respectively. This experiment contributes to the limited knowledge of acute sub-lethal effects of potential oil spills to the Arctic pelagic food web.

Effect of oil spill on compound-specific stable carbon isotope composition of amino acid in Nitzschia closterium and Heterosigma akashiwo

Marine microalgae, the most important primary producers in marine ecosystems, are susceptible to toxicants and usually selected as the promising organisms for chemical risk testing. The stable carbon isotope ratios (δ13C) of amino acids (AAs) were measured to determine if compound-specific stable isotope analysis could be used to provide insight into the toxic effects of oil spill on the marine microalgae. The water accommodated fractions (WAF) of #180 fuel oil were selected as the toxic targets with different concentrations in acute (96 h) toxic tests. Naphthalene, phenanthrene, anthracene, and fluoranthene were detected as the predominant congeners of polycyclic aromatic hydrocarbons in the WAF, and may be the main toxic components. At the same WAF concentration, the δ13C values of leucine in Nitzschia closterium increased with the culture time, whereas decreased in Heterosigma akashiwo. However, with the increasing WAF concentrations, the δ13C values of glutamic acid exhibited the increasing trend in both of the two kinds of microalgae at the same culture time. The similar phenomenon was also observed for histidine in H. akashiwo, while opposite in N. closterium, but both enriching in δ13C compared to the controlled experiments without the WAF. Under the different culture times and WAF concentrations, the δ13C values of alanine and methionine showed unexpected trend. These findings indicated that the synthesis pathways of AA in microalgae cells were affected by the WAF, thus leading to the carbon isotopic fractionation in AAs. The present study provide a novel approach to explore the toxicity mechanism of primary producers under oil spill stress, and monitor and evaluate the marine ecological risk of oil spill pollution.

Effect of oil spill stress on fatty acid stable carbon isotope composition of Ulva pertusa

Petroleum is one of the most important pollutants in the marine ecosystem, and oil spills have a long-term effect on the marine environment. After an oil spill, petroleum continues to diffuse, flowing to the intertidal zone where it accumulates. Ulva pertusa is the main macroalgae species in the intertidal zone of the ocean, and petroleum is toxic to marine macroalgae. Fatty acids, which are involved in various physiological activities, including metabolism, are indispensable substances in organisms. Thus, an organism's fatty acid composition reflects its physiological state. Because fatty acids are carbon-rich compounds, their stable carbon isotope composition can be measured to assess the state of the organism. In this study, the effects of the water accommodated fraction (WAF) of different concentrations of 180# fuel oil and 0# diesel oil on the chlorophyll a and fatty acid carbon stable isotope composition (δ13CFAs) of U. pertusa were assessed. The fatty acid carbon stable isotope value varied among the different petroleum treatments, and high WAF concentrations significantly inhibited the synthesis of unsaturated fatty acids. Thus, exposure to petroleum products affected the synthesis of fatty acids in U. pertusa and also caused carbon stable isotope fractionation during the fatty acid synthesis process. The results indicate that the saturated fatty acid δ13C16:0 and the unsaturated fatty acid δ13C18:1 can be used as biomarkers to evaluate the degree of petroleum-induced stress in U. pertusa and that carbon stable isotope analysis of U. pertusa can be used as an environmental risk assessment tool in the intertidal zone.

Chemically-dispersed crude oil and dispersant affects sperm fertilizing ability, but not sperm swimming behaviour in capelin (Mallotus villosus)

The effects of petroleum aromatic hydrocarbons (PAHs) on the embryonic and larval life stages of teleosts have been extensively examined. However, very little work has been conducted on how spilled oil affects fish sperm and there is no related knowledge concerning oil dispersing agents. The objective of our study was to determine sperm performance of a teleost fish under direct exposure to different concentrations of WAF (water accommodated fraction) and CEWAF (chemically enhanced water accommodated fraction). Capelin sperm motility, swimming behaviour, and sperm fertilization ability were evaluated in a scenario of an oil spill untreated (WAF) and treated (CEWAF) with the dispersant Corexit® EC9500A. Sperm fertilizing ability was lower when exposed to CEWAF concentrations of 16.1 × 103 μg/L total petroleum hydrocarbons and 47.9 μg/L PAH, and when exposed to the dispersant alone. The mechanism responsible for this reduced fertilizing ability is not clear. However, it is not related to the percentage of motile sperm or sperm swimming behaviour, as these were unaffected. WAF did not alter sperm swimming characteristics nor the fertilizing ability. We suggest the dispersant rather than the dispersed oil is responsible for the decrease in the sperm fertilizing ability and hypothesize that the surfactants present in the dispersant affect sperm membrane functionality.

A method for the production of large volumes of WAF and CEWAF for dosing mesocosms to understand marine oil snow formation

Marine oil snow (MOS) formation is a mechanism to transport oil from the ocean surface to sediments. We describe here the use of 110L mesocosms designed to mimic oceanic parameters during an oil spill including the use of chemical dispersants in order to understand the processes controlling MOS formation. These experiments were not designed to be toxicity tests but rather to illustrate mechanisms. This paper focuses on the development of protocols needed to conduct experiments under environmentally relevant conditions to examine marine snow and MOS. The experiments required the production of over 500 liters of water accommodated fraction (WAF), chemically enhanced water accommodated fraction of oil (CEWAF) as well as diluted CEWAF (DCEWAF). A redesigned baffled (170 L) recirculating tank (BRT) system was used. Two mesocosm experiments (M1 and M2) were run for several days each. In both M1 and M2, marine snow and MOS was formed in controls and all treatments respectively. Estimated oil equivalent (EOE) concentrations of CEWAF were in the high range of concentrations reported during spills and field tests, while WAF and DCEWAF concentrations were within the range of concentrations reported during oil spills. EOE decreased rapidly within days in agreement with historic data and experiments.

Characterization of dissolved and particulate phases of water accommodated fractions used to conduct aquatic toxicity testing in support of the Deepwater Horizon natural resource damage assessment.

In response to the Deepwater Horizon oil spill, the Natural Resource Trustees implemented a toxicity testing program that included 4 different Deepwater Horizon oils that ranged from fresh to weathered, and 3 different oil-in-water preparation methods (including one that used the chemical dispersant Corexit 9500) to prepare a total of 12 chemically unique water accommodated fractions (WAFs). We determined how the different WAF preparation methods, WAF concentrations, and oil types influenced the chemical composition and concentration of polycyclic aromatic hydrocarbons (PAHs) in the dissolved and particulate phases over time periods used in standard toxicity tests. In WAFs prepared with the same starting oil and oil-to-water ratio, the composition and concentration of the dissolved fractions were similar across all preparation methods. However, these similarities diverged when dilutions of the 3 WAF methods were compared. In WAFs containing oil droplets, we found that the dissolved phase was a small fraction of the total PAH concentration for the high-concentration stock WAFs; however, the dissolved phase became the dominant fraction when it was diluted to lower concentrations. Furthermore, decreases in concentration over time were mainly related to surfacing of the larger oil droplets. The initial mean diameters of the droplets were approximately 5 to 10 μm, with a few droplets larger than 30 μm. After 96 h, the mean droplet size decreased to 3 to 5 μm, with generally all droplets larger than 10 μm resurfacing. These data provide a detailed assessment of the concentration and form (dissolved vs particulate) of the PAHs in our WAF exposures, measurements that are important for determining the effects of oil on aquatic species. Environ Toxicol Chem 2017;36:1460-1472. © 2017 SETAC.

Toxicity assessment of water-accommodated fractions from two different oils using a zebrafish (Danio rerio) embryo-larval bioassay with a multilevel approach

Petroleum compounds from chronic discharges and oil spills represent an important source of environmental pollution. To better understand the deleterious effects of these compounds, the toxicity of water-accommodated fractions (WAF) from two different oils (brut Arabian Light and Erika heavy fuel oils) were used in this study. Zebrafish embryos (Danio rerio) were exposed during 96h at three WAF concentrations (1, 10 and 100% for Arabian Light and 10, 50 and 100% for Erika) in order to cover a wide range of polycyclic aromatic hydrocarbon (PAH) concentrations, representative of the levels found after environmental oil spills. Several endpoints were recorded at different levels of biological organization, including lethal endpoints, morphological abnormalities, photomotor behavioral responses, cardiac activity, DNA damage and exposure level measurements (EROD activity, cyp1a and PAH metabolites). Neither morphological nor behavioral or physiological alterations were observed after exposure to Arabian Light fractions. In contrast, the Erika fractions led a high degree of toxicity in early life stages of zebrafish. Despite of defense mechanisms induced by oil, acute toxic effects have been recorded including mortality, delayed hatching, high rates of developmental abnormalities, disrupted locomotor activity and cardiac failures at the highest PAH concentrations (∑TPAHs=257,029±47,231ng·L−1). Such differences in toxicity are likely related to the oil composition. The use of developing zebrafish is a good tool to identify wide range of detrimental effects and elucidate their underlying foundations. Our work highlights once more, the cardiotoxic action (and potentially neurotoxic) of petroleum-related PAHs.

Whole Spectrum of Cytochrome P450 Genes and Molecular Responses to Water-Accommodated Fractions Exposure in the Marine Medaka

Water-accommodated fractions (WAFs) of crude oil include chemicals that are potent toxicants in fish. Increasing oil pollution thus demands a better understanding of molecular mechanisms for detoxification, metabolism, toxicity, and adaptation of fish. Previous studies with fish show modulation of expression of key genes in relation to stress response against WAF exposure, but there is still a lack of studies on responses of cytochrome P450 (CYP) genes and changes in biotransformation upon WAF exposure. In this study, we used the full spectrum of CYP genes of the marine medaka, Oryzias melastigma, to understand their potential mode of action on WAFs-triggered molecular mechanisms. We also analyzed further CYP-involved detoxification and endogenous steroidogenic metabolism after exposure to different concentrations of WAFs over different time courses in the marine medaka. Also, detoxification- and antioxidant-related enzymes' activities were analyzed with different concentrations of WAFs. As a result, the WAF exposure induced CYP-involved detoxification mechanism but reduced CYP-involved steroidogenic metabolism in the marine medaka. These data suggest that whole CYP profiling can be a way of understanding and uncovering the mode of action particularly with respect to emerging chemicals such as WAF exposure with the new finding that WAFs have dual functions on CYP-involved metabolisms.

Acute and chronic toxicity study of the water accommodated fraction (WAF), chemically enhanced WAF (CEWAF) of crude oil and dispersant in the rock pool copepod Tigriopus japonicus

We determined the toxicity of the water accommodated hydrocarbon fraction (WAF), two chemically enhanced WAFs (CEWAFs; CEWAF-C, Crude oil+Corexit 9500 and CEWAF-H, Crude oil+Hiclean) of crude oil and two dispersants (Corexit 9500 and Hiclean) to the rock pool copepod Tigriopus japonicus. In the acute toxicity test, Corexit 9500 was the most toxic of all the chemicals studied. The nauplius stage of T. japonicus was more susceptible to the toxic chemicals studied than the adult female. The toxicity data using the nauplius stage was then considered as baseline to determine the spiking concentration of chemicals for chronic toxicity tests on the copepod. As the endpoints in the chronic toxicity test, survival, sex ratio, developmental time and fecundity of the copepod were used. All chemicals used in this study resulted in increased toxicity in the F1 generation. The lowest-observed-adverse-effect (LOAE) concentrations of WAF, CEWAF-H, CEWAF-C, Hiclean and Corexit 9500 were observed to be 50%, 10%, 0.1%, 1% and 1%, respectively. The results in present study imply that copepods in marine may be negatively influenced by spilled oil and dispersant.

Evaluation of biomarker potential of cytochrome P450 1A (CYP1A) gene in the marine medaka, Oryzias melastigma exposed to water-accommodated fractions (WAFs) of Iranian crude oil

CYP1A is involved in the metabolism of diverse chemicals, including polycyclic aromatic hydrocarbons and alkylated-PAHs, as a first line of detoxification mechanism. First, we identified and characterized the CYP1A gene from the marine medaka, Oryzias melastigma. O. melastigma CYP1A (Om-CYP1A) showed a high similarity of motifs/domains compared to those of vertebrates in their amino acid sequences. To check whether the Om-CYP1A would be inducible, we tested two strong CYP1A inducers, β-naphthoflavone (β-NF) and benzo[α]pyrene (B[α]P), and observed concentration-dependent transient expression on transcripts of Om-CYP1A for 96h over a wide range of concentrations. Om-CYP1A mRNA level was significantly increased in exposure to different concentrations of β-NF and B[α]P, and its expression was highly transcribed within 12h upon the exposure to low concentrations of both chemicals. Inducible transcript profiles revealed that Om-CYP1A would be associated with the toxicant metabolism via AhREs/DREs/XREs in its promoter region. To uncover the effects of the water-accommodated fraction (WAF) of crude oil on transcripts of Om-CYP1A, we measured mRNA expression of Om-CYP1A towards different concentrations of WAF for 24h. As a result, WAF exposure significantly increased Om-CYP1A transcripts at all concentrations as well as during time-course experiments for 96h. In this paper, we demonstrated that WAF would trigger up-regulation of the CYP1A gene that would be associated with the initiation of the cellular defense systems. This finding provides a better understanding of the molecular mechanism of cellular protection particularly that involved in the WAF-mediated cellular response in O. melastigma.

Effects of Polycyclic Aromatic Hydrocarbons on Sexual Maturity of Atlantic Cod, Gadus morhua, following Chronic Exposure

Mature Atlantic cod, Gadus morhua , were exposed in an ambient flow-through sea water system to low concentrations of water-accommodated fractions (WAFs) of a crude oil for periods of 38-92 days to ascertain its effects on sexual maturity. Some exposed groups were depurated after exposure for 4 to 287 days. Minimal mortality but a greater percentage of skin and gill lesions and a reduction in weight gained, especially in females, occurred in the WAF-exposed cod in contrast to controls in summer-autumn. The WAFs also disrupted gonadal development and delayed spawning and spermiation although the fish were fed to satiation. Sexual development was also interrupted following exposure to the WAFs and depuration afterwards. These results suggest that polycyclic aromatic hydrocarbons in the water column can potentially disrupt reproduction in Atlantic cod and eventually affect inshore populations inhabiting the cold subarctic waters in Newfoundland and Labrador in event of a major oil spill from offshore wells or supertankers.

Lung epithelial cell death induced by oil-dispersant mixtures

The dispersants used in oil spill disasters are claimed to be safe, but increased solubility of high-molecular-weight components in crude oil is of public health concern. The water-accommodated fractions (WAF) of crude oil mixed with dispersants may become airborne and cause lung epithelial damage when inhaled. This study was designed to examine the cell death and related death pathways of lung epithelial cells in response to WAF. Cultured A549 cells were treated for 2 or 24h with different concentrations of WAF. The WAF was prepared by mixing each of the dispersants (Corexit EC9527A, Corexit EC9500A and Corexit EC9580A) with crude oil for extraction with PBS. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide MTT assay, lactate dehydrogenase assay, morphology and cleaved caspase 9 protein, and microtubule-associated protein 1 light chain 3 were all used to measure cell viability, necrosis, apoptosis and autophagy quantitation, respectively. Results showed that the WAF of oil-dispersant mixtures caused cell death in the lung epithelial cells, in a dose-dependent manner, with the major cellular pathways of necrosis and apoptosis involved. Autophagy also occurred in cells exposed to WAF mixtures at lower concentrations before any detectable cell death, indicating greater sensitivity to WAF exposure. The three types of cell behavior, namely necrosis, apoptosis and autophagy, may play different roles in oil spill-related respiratory disorders.