The influence of temperature in sea urchin embryo toxicity of crude and bunker oils alone and mixed with dispersant.

This investigation deals with how temperature influences oil toxicity, alone or combined with dispersant (D). Larval lengthening, abnormalities, developmental disruption, and genotoxicity were determined in sea urchin embryos for assessing toxicity of low-energy water accommodated fractions (LEWAF) of three oils (NNA crude oil, marine gas oil -MGO-, and IFO 180 fuel oil) produced at 5-25 °C. PAH levels were similar amongst LEWAFs but PAH profiles varied with oil and production temperature. The sum of PAHs was higher in oil-dispersant LEWAFs than in oil LEWAFs, most remarkably at low production temperatures in the cases of NNA and MGO. Genotoxicity, enhanced after dispersant application, varied depending on the LEWAF production temperature in a different way for each oil. Impaired lengthening, abnormalities and developmental disruption were recorded, the severity of the effects varying with oil, dispersant application and LEWAF production temperature. Toxicity, only partially attributed to individual PAHs, was higher at lower LEWAF production temperatures.

Development of an analytical method for the simultaneous determination of 50 semi-volatile organic contaminants in wastewaters.

This work describes the development of a robust analytical methodology for the simultaneous determination of 50 semi-volatile organic compounds (SVOCs) in wastewater effluent samples by solid-phase extraction (SPE) followed by gas chromatography coupled to mass spectrometry (GC-MS) analysis. In this work, we studied extensively whether the validated SPE method used for the analysis of polar compounds in wastewaters could be extended to the analysis of non-polar compounds in the same analytical run. To that aim, the effect of different organic solvents in the SPE process (i.e., sample conditioning prior to SPE, elution solvent and evaporation steps) was evaluated. In this sense, the addition of methanol to wastewater samples before the extraction, the use of hexane:toluene (4:1, v/v) mixture for the quantitative elution of target compounds, and the addition of isooctane during the evaporation were required to minimize analyte losses during SPE and enhance extraction yields. Overall, the developed methodology showed a good performance for the determination of 50 SVOCs, and was further applied to the analysis of real wastewater effluent samples.•A validated SPE method for polar compounds was extended to the analysis of non-polar compounds.•Elution with hex:tol (4:1, v/v) and the addition of isooctane during the evaporation yield good recoveries.•The developed methodology was suitable for the determination of 50 SVOCs in aqueous samples.

Application of a biological multilevel response approach in the copepod Acartia tonsa for toxicity testing of three oil Water Accommodated Fractions.

Copepods play a critical role in the marine food webs, being a food source for marine organisms. In this study, we investigated the toxic effects of Water Accommodated Fractions (WAFs) from three types of oil: Naphthenic North Sea crude oil (NNS), Intermediate Fuel Oil (IFO 180) and a commercial Marine Gas Oil (MGO). The WAFs were prepared at 10 °C and 30 PSU (practical salinity unit), and tested on the marine copepod Acartia tonsa at different endpoints and at different levels of biological organization. We determined the median lethal concentrations after 96 h (LC50) and reproduction capabilities were calculated in adult females following seven days of exposure to sublethal WAF doses. The total lipid content was measured in reproductive females using Nile red lipophilic dye after 96 h of WAF exposure. We also measured the transcription levels of genes involved in antioxidant response and xenobiotic biotransformation after short exposure for 48 h. High doses (7% WAF) of MGO affected survival, percentage of fecund females, egg hatching success, and total lipid content. The IFO 180 WAF affected, at medium (20%) and high (40%) doses, the number of fecund females, mortality and produced significant effects on gene expression levels. In conclusion, toxicity assays showed that the WAFs prepared from refined oils were more toxic than crude oil WAF to Acartia tonsa.

In vitro toxicity testing in hemocytes of the marine mussel Mytilus galloprovincialis (L.) to uncover mechanisms of action of the water accommodated fraction (WAF) of a naphthenic North Sea crude oil without and with dispersant.

Dispersants used in oil spills could result toxic to marine organisms and could influence the toxicity of oil compounds. The aim of this work was to uncover the mechanisms of action of the water accommodated fraction (WAF) of a naphthenic North Sea crude oil produced at 10, 15 and 20 °C without and with the dispersant Finasol OSR52 (WAF and WAFD, respectively) using hemocytes of the marine mussel Mytilus galloprovincialis. Primary cultures of hemocytes were exposed in glass-coated microplates to different WAF or WAFD dilutions (0.25, 2.5, 25, 50 and 100%) and to the dispersant alone at the same concentrations present in the WAFD dilutions (1.25, 12.5, 125, 250 and 500 mg/L). Of the two in vitro approaches tested, the second one was selected which involved exposure of hemocytes for 4 h to unfiltered WAF, WAFD and dispersant dilutions without cell culture media. WAF decreased hemocytes viability only at the highest dilution whereas WAFD and the dispersant alone were cytotoxic at the three highest concentrations. Temperature of production of WAF, WAFD and dispersant did not influence their cytotoxicity to hemocytes. WAF increased ROS production and MXR transport activity in hemocytes. Exposure to WAFD and dispersant increased ROS production, provoked plasma membrane and actin cytoskeleton disruption and decreased phagocytic activity. In conclusion, the dispersant tested was toxic to mussel hemocytes and it greatly increased the toxicity of WAFD. The present data could be useful for the environmental risk assessment of oil spills and their remediation strategies in the marine environment.

Influence of season-depending ecological variables on biomarker baseline levels in mussels (Mytilus trossulus) from two Baltic Sea subregions.

For reliable mussel monitoring programmes based on biomarkers, regionally relevant reference values and their natural variability need to be known. The Baltic Sea exhibits high inter-regional and seasonal variability in physical factors such as salinity, temperature and primary production. The aim of this pilot study is to depict the effects of season-related environmental factors in a selected battery of biomarkers in two environmentally different subregions of the Baltic Sea to help establishing reference data for biochemical, cellular and tissue-level biomarkers. In order to achieve that, mussels were collected from reference sites in Kiel (Germany) and Tvärminne (Finland) during three seasons: summer and autumn 2016, and spring 2017. Finally, in order to characterize the ecological situation, analysis of the chemical tissue burden was performed and chlorophyll­a and particulate organic carbon concentration and temperature changes were analyzed at each sampling locality using satellite remote sensing images. An integrated biomarker response index was performed to summarize the biomarker responses of each locality and season. The biochemical endpoints showed seasonal variability regulated by temperature, food supply and reproductive cycle, while among the cellular endpoints only lipofuscin accumulation and lysosomal structural changes showed slight seasonal variation. Seasonal changes in tissue level biomarkers were observed only at the northern Baltic Sea site Tvärminne, dictated by the demanding energetic trade-off caused by reproduction. In conclusion, the characterization of the ecological variables and physico-chemical conditions at each site, is crucial to perform a reliable assessment of the effects of a hypothetical pollution scenario in the Baltic Sea. Moreover, reference levels of biomarkers and their responses to natural environmental conditions must be established.