Combined effects of high hydrostatic pressure and dispersed oil on the metabolism and the mortality of turbot hepatocytes (Scophthalmus maximus).

Since the DeepWater Horizon oil spill and the use at 1450 m depth of dispersant as a technical response, the need of relevant ecotoxicological data on deep-sea ecosystems becomes crucial. In this context, this study focused on the effect of high hydrostatic pressure (10.1 MPa) on turbot hepatocytes isolated from fish exposed either to chemically dispersed oil, mechanically dispersed oil or dispersant alone. Potential combined effects of oil/dispersant and hydrostatic pressure, were assessed on cell mortality (total cell death, necrosis and apoptosis), cell viability and on hepatocyte oxygen consumption (MO2). No change in cell mortality was observed in any of the experimental conditions, whereas, the results of cell viability showed a strong and significant increase in the two oil groups independently of the pressure exposure. Finally, oil exposure and hydrostatic pressure have additive effects on oxygen consumption at a cellular level. Presence of dispersant prevent any MO2 increase in our experimental conditions. These mechanistic effects leading to this increased energetic demand and its eventual inhibition by dispersant must be investigated in further experiments.

Deep-sea versus shallow conditions: a comparative ecobarotoxicological study.

In the context of new oil exploration/production areas, knowledge of the biological impact of dispersed oil in the deep-sea environment is essential. Hence, the aim of this study was to perform a comparison, at atmospheric pressure (0.1 MPa) and at a high hydrostatic pressure corresponding to 1000 m depth (10.1 MPa), of lethal concentrations (LC) on a model fish, Scophthalmus maximus, exposed to chemically dispersed oil. Fish were exposed concomitantly at 0.1 and 10.1 MPa using two exposure tanks connected to the same source tank thanks to a closed circuit. Acute toxicity was evaluated at 24 h through the determination of LC10 and LC50 (respectively, 10 and 50% of mortality) calculated from measured total petroleum hydrocarbon concentrations in the water. No statistical differences were observed between the LC10 at 0.1 MPa (46.1 mg L- 1) and the LC10 at 10.1 MPa (31.0 mg L- 1), whereas the LC50 of fish exposed to 0.1 MPa (90.8 mg L- 1) was significantly higher than the LC50 at 10.1 MPa (50.9 mg L- 1). These results clearly show an increase in oil toxicity under high hydrostatic pressure. This effect may be due to synergistic effects of pressure and oil contamination on fish energetic metabolism.

Assessing the long-term effect of exposure to dispersant-treated oil on fish health using hypoxia tolerance and temperature susceptibility as ecologically relevant biomarkers.

The ecological and economic importance of fish act as a brake on the development of chemical dispersants as operational instruments following oil spills. Although a valuable and consistent body of knowledge exists, its use in spill response is limited. The objective of the present study was to increase current knowledge base to facilitate the translation of published data into information of operational value. Thus we investigated the dose-response relationship between dispersant-treated oil exposure and ecologically relevant consequences by combining laboratory and field experiments. Effects were examined over almost a year using juveniles of the slowly growing, commercially important European sea bass (Dicentrarchus labrax). A reliable interpretation of biomarker responses requires a complete knowledge of the factors likely to affect them. Interpopulational variability is of particular importance in environmental impact assessment because biomarker responses from a population collected in an impacted area are classically compared with those collected in a clean site. Our study revealed no effect of the exposure to dispersant-treated oil on fish hypoxia tolerance and temperature susceptibility at 1 and 11 mo post exposure. Similarly, no effect of the exposure was observed on the ability of the fish to cope with environmental contingencies in the field, regardless of the dose tested. Thus we feel confident to suggest that a 48-h exposure to chemically treated oil does not affect the ability of sea bass to cope with mild environmental contingencies. Finally, investigation of interpopulation variability revealed large differences in both hypoxia tolerance and temperature susceptibility among the 2 populations tested, suggesting that this variability may blur the interpretation of population comparisons as classically practiced in impact assessment. Environ Toxicol Chem 2019;38:210-221. © 2018 SETAC.

Avoidance threshold to oil water-soluble fraction by a juvenile marine teleost fish.

When oil spills occur, behavior is the first line of defense for a fish to avoid being contaminated. We determined the avoidance threshold of the European seabass (Dicentrarchus labrax) to the water-soluble fraction (WSF) of oil using a dual-flow choice box. The results showed that a plume of 20%-diluted WSF (total polycyclic aromatic hydrocarbon [PAH] concentration: 8.54 µg L-1 ) triggered a significant avoidance response that was detected within 7.5 min of introducing WSF-contaminated water into the experimental setup. However, the ecological relevance of seabass capacity to detect and avoid WSF remains to be established. In the short term, such a response is indeed liable to reduce seabass contact time with oil-contaminated water and thus preserve their functional integrity. In the long term, however, avoidance may contribute to the displacement of a population into a possibly less auspicious environment, with consequences very similar to those of contaminant exposure, that is, disturbed population dynamics and demography. Environ Toxicol Chem 2018;37:854-859. © 2017 SETAC.

Dispersed oil decreases the ability of a model fish (Dicentrarchus labrax) to cope with hydrostatic pressure.

Data on the biological impact of oil dispersion in deep-sea environment are scarce. Hence, the aim of this study was to evaluate the potential interest of a pressure challenge as a new experimental approach for the assessment of consequences of chemically dispersed oil, followed by a high hydrostatic pressure challenge. This work was conducted on a model fish: juvenile Dicentrarchus labrax. Seabass were exposed for 48 h to dispersant alone (nominal concentration (NC) = 4 mg L-1), mechanically dispersed oil (NC = 80 mg L-1), two chemically dispersed types of oil (NC = 50 and 80 mg L-1 with a dispersant/oil ratio of 1/20), or kept in clean seawater. Fish were then exposed for 30 min at a simulated depth of 1350 m, corresponding to pressure of 136 absolute atmospheres (ATA). The probability of fish exhibiting normal activity after the pressure challenge significantly increased from 0.40 to 0.55 when they were exposed to the dispersant but decreased to 0.26 and 0.11 in the case of chemical dispersion of oil (at 50 and 80 mg L-1, respectively). The chemical dispersion at 80 mg L-1 also induced an increase in probability of death after the pressure challenge (from 0.08 to 0.26). This study clearly demonstrates the ability of a pressure challenge test to give evidence of the effects of a contaminant on the capacity of fish to face hydrostatic pressure. It opens new perspectives on the analysis of the biological impact of chemical dispersion of oil at depth, especially on marine species performing vertical migrations.

Sensitivity of the deep-sea amphipod Eurythenes gryllus to chemically dispersed oil.

In the context of an oil spill accident and the following oil spill response, much attention is given to the use of dispersants. Dispersants are used to disperse an oil slick from the sea surface into the water column generating a cloud of dispersed oil droplets. The main consequence is an increasing of the sea water-oil interface which induces an increase of the oil biodegradation. Hence, the use of dispersants can be effective in preventing oiling of sensitive coastal environments. Also, in case of an oil blowout from the seabed, subsea injection of dispersants may offer some benefits compared to containment and recovery of the oil or in situ burning operation at the sea surface. However, biological effects of dispersed oil are poorly understood for deep-sea species. Most effects studies on dispersed oil and also other oil-related compounds have been focusing on more shallow water species. This is the first approach to assess the sensitivity of a macro-benthic deep-sea organism to dispersed oil. This paper describes a toxicity test which was performed on the macro-benthic deep-sea amphipod (Eurythenes gryllus) to determine the concentration causing lethality to 50% of test individuals (LC50) after an exposure to dispersed Brut Arabian Light (BAL) oil. The LC50 (24 h) was 101 and 24 mg L(-1) after 72 h and 12 mg L(-1) at 96 h. Based on EPA scale of toxicity categories to aquatic organisms, an LC50 (96 h) of 12 mg L(-1) indicates that the dispersed oil was slightly to moderately toxic to E. gryllus. As an attempt to compare our results to others, a literature study was performed. Due to limited amount of data available for dispersed oil and amphipods, information on other crustacean species and other oil-related compounds was also collected. Only one study on dispersed oil and amphipods was found, the LC50 value in this study was similar to the LC50 value of E. gryllus in our study. Since toxicity data are important input to risk assessment and net environmental benefit analyses, and since such data are generally lacking on deep-sea species, the data set produced in this study is of interest to the industry, stakeholders, environmental management, and ecotoxicologists. However, studies including more deep-sea species covering different functional groups are needed to evaluate the sensitivity of the deep-sea compartments to dispersed oil relative to other environmental compartments.

Application of the Activity Framework for Assessing Aquatic Ecotoxicology Data for Organic Chemicals.

Toxicological research in the 1930s gave the first indications of the link between narcotic toxicity and the chemical activity of organic chemicals. More recently, chemical activity has been proposed as a novel exposure parameter that describes the fraction of saturation and that quantifies the potential for partitioning and diffusive uptake. In the present study, more than 2000 acute and chronic algal, aquatic invertebrates and fish toxicity data, as well as water solubility and melting point values, were collected from a series of sources. The data were critically reviewed and grouped by mode of action (MoA). We considered 660 toxicity data to be of acceptable quality. The 328 data which applied to the 72 substances identified as MoA 1 were then evaluated within the activity-toxicity framework: EC50 and LC50 values for all three taxa correlated generally well with (subcooled) liquid solubilities. Acute toxicity was typically exerted within the chemical activity range of 0.01-0.1, whereas chronic toxicity was exerted in the range of 0.001-0.01. These results confirm that chemical activity has the potential to contribute to the determination, interpretation and prediction of toxicity to aquatic organisms. It also has the potential to enhance regulation of organic chemicals by linking results from laboratory tests, monitoring and modeling programs. The framework can provide an additional line of evidence for assessing aquatic toxicity, for improving the design of toxicity tests, reducing animal usage and addressing chemical mixtures.

Growth and immune system performance to assess the effect of dispersed oil on juvenile sea bass (Dicentrarchus labrax).

The potential impact of chemically and mechanically dispersed oil was assessed in a model fish of European coastal waters, the sea bass Dicentrarchus labrax. Juvenile sea bass were exposed for 48h to dispersed oil (mechanically and chemically) or dispersants alone. The impact of these exposure conditions was assessed using growth and immunity. The increase observed in polycyclic aromatic hydrocarbon metabolites in bile indicated oil contamination in the fish exposed to chemical and mechanical dispersion of oil without any significant difference between these two groups. After 28 days of exposure, no significant differences were observed in specific growth rate,apparent food conversion efficiency and daily feeding). Following the oil exposure, fish immunity was assessed by a challenge with Viral Nervous Necrosis Virus (VNNV). Fish mortality was observed over a 42 day period. After 12 days post-infection, cumulative mortality was significantly different between the control group (16% p≤0.05) and the group exposed to chemical dispersion of oil (30% p≤0.05). However, at the end of the experiment, no significant difference was recorded in cumulative mortality or in VNNV antibodies secreted in fish in responses to the treatments. These data suggested that in our experimental condition, following the oil exposure, sea bass growth was not affected whereas an impact on immunity was observed during the first days. However, this effect on the immune system did not persist over time.

Innate immunity and antioxidant systems in different tissues of sea bass (Dicentrarchus labrax) exposed to crude oil dispersed mechanically or chemically with Corexit 9500.

The aim of the study was to evaluate effects of chemically dispersed oil by the dispersant Corexit 9500 on innate immunity and redox defenses in a marine model fish. Sea bass (Dicentrarchus labrax) were exposed 48h to four experimental conditions: a control group (C), a group only exposed to the dispersant (D; 3.6mg/L) and two groups exposed to 80mg/L oil mechanically or chemically dispersed (MD; CD). Alternative pathway of complement activity and lysozyme concentration was measured in plasma in order to evaluate the general fish health status. Total glutathione, glutathione peroxidase (GPX) and superoxide dismutase (SOD) were analyzed in gills, liver, brain, intestine and muscle. The chemical dispersion induced a significant reduction of lysozyme concentration when compared to the controls, and the hemolytic activity of the alternative complement pathway was increased in mechanical and chemical dispersion. The analysis of SOD, GPX and total glutathione showed that antioxidant defenses were activated in liver and reduced in intestine and brain. Dispersant was also responsible for an SOD activity inhibition in these two last tissues, demonstrating a direct effect of this dispersant on reactive oxygen species homeostasis that can be interpreted as a signal of tissue toxicity. This result should raise concern about the use of dispersants and show that they can lead to adverse effects on marine species.

Effect of dispersed crude oil on cardiac function in seabass Dicentrarchus labrax.

In this study, the impact of dispersed oil was assessed in Dicentrarchus labrax, a fish frequently used as an oil contamination indicator species. Fish were exposed for 48h to (mechanically and chemically) dispersed oil and dispersant alone. The impact of these exposure conditions was assessed on cardiac function by measuring (i) the contraction strength, the contraction and the relaxation speeds (ii) the cardiac energy metabolism using respirometry on permeabilized cardiac fibers. Compared to control, the increase of polycyclic aromatic metabolites observed in the bile indicated oil contamination in our fish. Following 48h of oil exposure at realistic oil concentrations, alterations of cardiac performances were observed. A decrease in contraction strength, contraction and relaxation speeds was observed in the presence of oil without effect of dispersant on these three parameters. Looking at cardiac energy metabolism, dispersant alone decreases all the activity of the respiratory chain and increases the proton leak. From these results, it appears that the observed decrease in cardiac performance in fish exposed to oil was not linked to a decrease in energy availability.

Acute toxicity of chemically and mechanically dispersed crude oil to juvenile sea bass (Dicentrarchus labrax): Absence of synergistic effects between oil and dispersants.

The goal of the present experiment was to assess the relative acute toxicities of mechanically and chemically dispersed oil (crude Arabian Light) in controlled conditions. Juvenile sea bass (Dicentrarchus labrax) were exposed to 4 commercial formulations of dispersants (Corexit EC9500A, Dasic Slickgone NS, Finasol OSR 52, Inipol IP 90), to mechanically dispersed oil, and to the corresponding chemical dispersions. Acute toxicity was evaluated at 24 h, 48 h, 72 h, and 96 h through the determination of 10%, 50%, and 90% lethal concentrations calculated from measured total petroleum hydrocarbon (TPH) concentrations; Kaplan-Meyer mortality analyses were based on nominal concentrations. Animals were exposed to the dissolved fraction of the oil and to the oil droplets (ranging from 14.0 µm to 42.3 µm for the chemical dispersions). Kaplan-Meyer analyses demonstrated an increased mortality in the case of chemical dispersions. This difference can be attributed mainly to differences in TPH, because the chemical lethal concentrations were not reduced compared with mechanical lethal concentrations (except after 24 h of exposure). The ratios of lethal concentrations of mechanical dispersions to the different chemical dispersions were calculated to allow direct comparisons of the relative toxicities of the dispersions. The results ranged from 0.27 to 3.59, with a mean ratio close to 1 (0.92). These results demonstrate an absence of synergistic effect between oil and chemical dispersants in an operational context.

Impact of dispersed fuel oil on cardiac mitochondrial function in polar cod Boreogadus saida.

In this study, impact of dispersed oil on cardiac mitochondrial function was assessed in a key species of Arctic marine ecosystem, the polar cod Boreogadus saida. Mature polar cod were exposed during 48 h to dispersed oil (mechanically and chemically) and dispersants alone. The increase observed in ethoxyresorufin-O-deethylase activity and polycyclic aromatic hydrocarbon metabolites in bile indicated no difference in contamination level between fish exposed to chemical or mechanical dispersion of oil. Oil induced alterations of O2 consumption of permeabilised cardiac fibres showing inhibitions of complexes I and IV of the respiratory chain. Oil did not induce any modification of mitochondrial proton leak. Dispersants did not induce alteration of mitochondrial activity and did not increase oil toxicity. These data suggest that oil exposure may limit the fitness of polar cod and consequently could lead to major disruption in the energy flow of polar ecosystem.

Photoselective vaporization of the prostate with GreenLight 120-W laser compared with monopolar transurethral resection of the prostate: a multicenter randomized controlled trial.

BACKGROUND: Evidence supporting the widespread use of GreenLight High Performance System (HPS) 120-W photoselective vaporization of the prostate (PVP) is lacking. OBJECTIVE: To assess the noninferiority of PVP compared with transurethral resection of the prostate (TURP) on urinary symptoms and the superiority of PVP over TURP on length of hospital stay. DESIGN, SETTING, AND PARTICIPANTS: A multicenter randomized controlled trial was conducted. INTERVENTION: Patients underwent monopolar TURP or PVP with the GreenLight HPS 120-W laser. MEASUREMENTS: International Prostate Symptom Score (IPSS), Euro-QOL questionnaire, uroflowmetry, Danish Prostate Symptom Score Sexual Function Questionnaire, sexual satisfaction, and adverse events were collected at 1, 3, 6, and 12 mo. The two groups were compared using the 95% confidence interval (CI) of median difference for testing noninferiority of the IPSS at 12 mo and the student t test for testing the difference in length of hospital stay. RESULTS AND LIMITATIONS: A total of 139 patients (70 vs 69 men in each group) were randomized. Median IPSS scores at 12-mo follow-up were 5 (interquartile range [IQR]: 3-8) for TURP versus 6 (IQR: 3-9) for PVP, and the 95% CI of the difference of the median was equal to -2 to 3. Because the upper limit of the 95% CI was >2 (the noninferiority margin), the hypothesis of noninferiority could not be considered demonstrated. Median length of stay was significantly shorter in the PVP group than in the TURP group, with a median of 1 (IQR: 1-2) versus 2.5 (IQR: 2-3.5), respectively (p<0.0001). Uroflowmetry parameters and complications were comparable in both groups. Sexual outcomes were slightly better in the PVP group without reaching statistical significance. CONCLUSIONS: The present study failed to demonstrate the noninferiority of 120-W GreenLight PVP versus TURP on prostate symptoms at 1 yr but showed that PVP was associated with a shorter length of stay in the hospital. TRIAL REGISTRATION: NCT01043588.

Diffracted wavefront measurement of a volume phase holographic grating at cryogenic temperature.

Flatness of the wavefront diffracted by grating can be mandatory for some applications. At ambient temperature, the wavefront diffracted by a volume phase holographic grating (VPHG) is well mastered by the manufacturing process and can be corrected or shaped by postpolishing. However, to be used in cooled infrared spectrometers, VPHGs have to stand and work properly at low temperatures. We present the measurement of the wavefront diffracted by a typical VPHG at various temperatures down to 150 K and at several thermal inhomogeneity amplitudes. The particular grating observed was produced using a dichromated gelatine technique and encapsulated between two glass blanks. Diffracted wavefront measurements show that the wavefront is extremely stable according to the temperature as long as the latter is homogeneous over the grating stack volume. Increasing the thermal inhomogeneity increases the wavefront error that pinpoints the importance of the final instrument thermal design. This concludes the dichromated gelatine VPHG technology, used more and more in visible spectrometers, can be applied as it is to cooled IR spectrometers.

Efficient syntheses of pterulone, pterulone B and related analogues.

An efficient synthesis of the three halogenated naturally occurring products, pterulone (2), pterulone B (3) and alcohol 5, and of a wide range of related unnatural analogues has been achieved starting from the two readily available 1-benzoxepine sulfonyl-containing intermediates 6a and 6b. The biological activities of pterulone and some of the synthesized analogues were tested against a wide spectrum of phytopathogenic fungi.

[Non-metastatic prostate cancer treated with exclusive radiotherapy: prognosis of patients according to lymph node radiologic or surgical assessment method. Multivariate retrospective analysis]. / Cancers de prostate non métastatiques traités par radiothérapie exclusive: devenir des patients selon la méthode d'évaluation ganglionnaire radiologique ou chirurgicale. Une analyse rétrospective multivariée.

PURPOSE: To compare retrospectively the outcome of localized prostate cancers treated by curative external radiotherapy in which the negative lymphatic status was either surgically or radiologically assessed. METHODS AND MATERIALS: From January 1986 to December 1995, 112 patients with localized prostate cancers were found to have no evidence of lymphatic disease in the pelvis. N0 status was assessed either surgically (61 patients, group pN-) or after a CT scan procedure (51 patients, group cN0). The treatment consisted of conventional external radiotherapy using a four-fields box technique to a total dose of 65 Gy. The pelvis was never irradiated. RESULTS: The two groups did not statistically differ according to age, PSA level, Gleason score, T stage and hormonal therapy. Actuarial NED survival rates were 80% and 60% at five and ten years respectively. At ten years, the actuarial NED survival rates were 78% and 34% in the pN- and cN0 groups respectively (p = 0.003). The multivariate analysis corroborated the positive impact of lymphatic dissection before radiotherapy on disease free survival of T1-T2 patients, but not for T3 stages. CONCLUSIONS: This retrospective study suggests the inability of CT scan to accurately evaluate the lymph node status in carcinoma of the prostate. Systematic ilio-obturator nodal dissection is strongly recommended in early stages before curative radiotherapy. Only pN-patients should be included in high dose conformal irradiation trials.