environment due to rainfall and surface runoff, and thus it may affect marine organisms. However, its toxicity and ecological risk to marine organisms remain largely unknown. Therefore, this study aims at investigating the environmental fate of larvicidal oil in the marine environment and its toxicities towards marine organisms at different trophic levels along the food chain. The composition of larvicidal oil was characterized by gas chromatography-mass spectrometry. It was found to consist mainly of aliphatic petroleum hydrocarbons (n-alkanes) ranging from n-octane to n-pentacosane. The concentrations of larvicidal oil were determined with the range from 6.92 mg/L to 53.89 mg/L., by analyzing water samples collected along coastal areas in Hong Kong. Standard acute toxicity tests were conducted to investigate their toxic effects to the marine microalgae
Isochrysis galbana and Chaetoceros gracilis (primary producers), the intertidal
copepod Tigriopus japonicas (a primary consumer), the brine shrimp Artemia
franciscana and fish embryos of the marine medaka Oryzias melastigma. Our
results showed that although all test marine species were not very sensitive to larvicidal oil with the ranking of their acute median lethal concentrations (LC50) that were all above the estimated hazardous concentration for 5% of species (HC5), the results of a probabilistic risk assessment showed that the local marine ecosystem had 65.7% of chance to be at risk (i.e., hazardous quotients > 1) from exposure to larvicidal oil using Monte Carlo simulation, indicating that the current risk was unacceptably high. Hence, monitoring and control on the use of larvicidal oil as mosquito control pesticide would be urgently needed to mitigate its ecological risks.
MO014
Effects of a coastal oil spill on marine invertebrates and their potential to recover
M.F. Lemos, S. Silva, Instituto Politécnico de Leiria / MARE IPLeiria There has been an increasing public concern and focus on marine contamination issues mainly due to the arising of emergent pollutants, posing a major threat to human and environmental health. Still, the contamination by polycyclic aromatic hydrocarbons (PAHs) remains one of the most ubiquitous sources of pollution in the marine environment, being reported to elicit toxic, carcinogenic and mutagenic effects on marine biota. Moreover, the assessment of these impacts in costal invertebrates after a spill, the extent of these effects and energetic trade-offs, potential recovery, and even which species to use is still deemed for an effective environmental contamination assessment. After an accidental industrial oil spill at the rocky shore of Peniche, Portugal in the summer of 2018, the water was analyzed during the low-tide for PAHs one week later and regularly throughout six months in the spilled beach and in 7 other rocky beaches in the vicinity. Also, at all locations,
Patella depressa and Gibbula umbilicalis organisms were collected, and several
biomarkers addressed. For both species, the neurotransmission enzyme acetylcholinesterase, oxidative stress enzyme catalase and superoxide dismutase, oxidative damage DNA damage and lipid peroxidation, energy metabolism lactate and isocitrate dehydrogenase, and electron transfer system, and carbohydrates, lipids and proteins energy reserves were assessed. The impacts of this oil spill over the two coastal invertebrate species’ biomarkers was compared over the
differentially PAH contaminated sites and their sensitivity evaluated. Also, organism’s ability to recover over time was also addressed and these tools and species potential for costal monitoring pollution scenarios discussed.
MO015
Effects of oil exposure on visual function in early life stage fishes
J.T. Magnuson, University of North Texas / Biology; A.J. Khursigara, The University of Texas at Austin / Marine Science Institute; E. Allmon, The University of Texas at Austin; A. Esbaugh, University of Texas Marine Science Institute / Department of Marine Science; R.M. Heuer, University of Miami / Marine Biology and Ecology; J.D. Stieglitz, M. Grosell, RSMAS University of Miami / Marine Biology and Ecology; A.P. Roberts, University of North Texas / Advanced Environmental Research Institute
The Deepwater Horizon oil spill released millions of barrels of oil into the Gulf of Mexico, coinciding with peak spawning periods of ecologically important fish species, such as the mahi-mahi (Coryphaena hippurus), red drum (Sciaenops
ocellatus), and sheepshead minnow (Cyprinodon variegatus). Downregulation of
genes important in eye development and function, as well as morphological abnormalities have resulted from polycyclic aromatic hydrocarbons (PAHs) present in the oil at concentrations less than 10 µg/L, impacting fish vision. Mahi-mahi, red drum, and sheepshead minnow embryos were exposed to weathered crude oil and assessed for visual function using the flicker-fusion principle to monitor an optomotor response, with subsequent histological analysis taken of each larvae’s retina. Oil-exposed larvae exhibited a reduced PAH-dependent optomotor response with a reduction in retinal layers and neuronal connections that play an important role in visual function and image processing. The present study provides evidence that weathered crude oil affects the visual system in developing larval fish, and relates oil-induced histological effects to behavioral endpoints. This research was made possible by a grant from The Gulf of Mexico Research Initiative. Grant No: SA-1520; Name: Relationship of Effects of Cardiac Outcomes in fish for Validation of Ecological Risk (RECOVER).
MO016
Effects of oil spill on coastal seaweed in the Arctic
S. Wegeberg, Aarhus University / Department of Bioscience; J. Fritt-Rasmussen, K. Gustavson, Aarhus University / Department of Bioscience - Arctic Environment In case of an acute oil spill response operation, decision making regarding the operational response strategy and prioritizing biology at risk must be resolute. For that a Net Environmental Benefit Analysis, NEBA, is often performed to achieve the optimal environmental benefit with respect to choice of oil spill combat methodology and biology at risk. To provide data for assessing beaching oil spill impacts in the Arctic areas, the effects of oil smothering of the macroalgae Fucus
distichus, which is a dominant species in the intertidal zone of the coasts in the
Arctic, as well as its self-cleaning potential by wash in sea, were studied. Effects of four different oil types were tested, including crude oil types, bunker oil and marine diesel. Different oil types have varying properties depending on the origin of crude oil and refinery process, and hence may have different effects due to their physical and chemical characterizations. Photosynthetic activity was measured as proxy for effect on growth and the self-cleaning potential was tested by wash in sea for oil smothered tips of F. distichus over a period of 2 weeks. The removal of the oils from the seaweed surface was considered as relatively fast (T1/2 ~ 3-4 days). Depending of oil type, the oil inhibited or stimulated photosynthetic activity. Marine diesel inhibited photosynthetic activity, whereas the three other oil types stimulated the activity. Thus, in general, the results indicated 1) that oil smothering was relatively fast washed off in the sea water; 2) that, depending on the oil type, photosynthetic activity were stimulated or inhibited; and 3) that the photosynthetic activity was still affected (stimulated or inhibited) even after 14 days, although oil on the tip surface was completely or almost completely washed off. The studies were funded by the European Commission Horizon 2020 programme and the Government of Greenland.
MO017
Effects of water accommodated fractions of crude oil on the Baltic Sea blue mussel Mytilus trossulus at different salinities
A. Ahvo, Finnisk Environment Institute / Marine Research Centre; R. Turja, Finnish Environment Institute, SYKE / Marine Research Centre; A. Reunamo, Finnish Environment Institute / Marine Research Centre; J. Nuutinen, Finnish Environment Institute / Laboratory Centre; K.K. Lehtonen, K.S. Jørgensen, Finnish Environment Institute / Marine Research Centre
In the Baltic Sea accidental oil spills are mainly combatted using mechanical collection. However, this method is insufficient in harsh weather conditions such as high waves or in the presence of ice. The use of dispersants is an alternative counteractive method but in the Baltic Sea their use is restricted by HELCOM recommendations since the chemically dispersed oil may cause severe toxic effects on marine biota. In addition, the behaviour of dispersants with oil under brackish water conditions is not well studied. In the present study, impacts of a crude oil and the dispersant Finasol 51 on marine biota were investigated under cold conditions (5ºC) at two salinities corresponding to the German (15.0) and southern Finnish coastal areas (5.6). Baltic Sea blue mussels (Mytilus trossulus) were exposed to the water accommodated fraction (WAF) and dispersed WAF (WAF-D) of naphthenic North Sea crude oil in a semi-static aquarium experiment. Concentration of WAF or WAF-D in the aquaria was 5%. The mussels were sampled after 0, 1, 7 and 21 days of exposure, and analyzed for accumulation of polycyclic aromatic hydrocarbons (PAHs), and biological effects including acetylcholinesterase,
glutathione-S-transferase, catalase and glutathione reductase activities, lipid peroxidation, and protein carbonylation. In addition, changes in Mytilus-associated bacterial communities extracted from the gills and digestive glands of the mussels were investigated by sequencing of 16S rRNA genes and quantitative PCR targeted to bacterial PAH-degradation genes. Water samples from the exposure aquaria were taken for oil and PAH analysis. Dispersant augmented the amount oil in the exposure water with 0.13 mg/l oil in 5.6 WAF compared to 44 mg/l oil in 5.6 WAF-D (GC-FID, petroleum hydrocarbons C10-C40). A significantly higher oil concentration was observed at the lower salinity WAF-D water with 44 mg/l oil at 5.6 and 1.82 mg/l oil at 15. The higher salinity and WAF-D elicited more oxidative stress and neurotoxic effects already after one day of exposure. Mytilus-associated bacterial communities also varied depending on salinity and the use of dispersant. The results indicate that during the application of dispersants salinity plays a key role both in regard to oil concentrations in water as well as biological effects observed in the exposed biota. This should be taken into careful consideration when designing oil spill mitigation procedures in the Baltic Sea.
MO018
Multiple biomarkers on the estuarine guppy Poecilia vivipara to monitor two brazilian tropical estuaries
A.G. Torreiro-Melo, UFPE Universidade Federal de Pernambuco / Department of Zoology; J.S. Silva, UFPE Universidade Federal de Pernambuco / Zoology; E. Zanardi-Lamardo, Universidade Federal de Pernambuco / Department of Oceanography; P.S. Carvalho, UFPE - Universidade Federal de Pernambuco / Zoología
Integrated approaches using biological responses in multiple organization levels are essential for environmental monitoring of tropical estuaries with ecologically relevant tools. The guppy Poecilia vivipara, native species with a broad tropical distribution, was utilized in such an approach, using in situ field exposures in cages
(IS) and resident individuals (RES) collected close to the mouth of two tropical estuaries, Bacia do Pina Estuarine System (BPES), and Barra de Jangada Estuarine System (BJES), in the Brazilian northeastern coast. This study is based on the analysis of water concentrations and internal accumulation of bile metabolites of polycyclic aromatic hydrocarbons (PAHs) by fixed fluorescence (FF), as well as biochemical responses related to the biotransformation of contaminants
ethoxyresorufin-O-deethylase (EROD) and glutathione S-transferase (GST), and to neurotransmission acetylcholinesterase (AChE). Behavioral activities related to swimming speed and resistance were also evaluated. Individuals grown in the laboratory were used for in situ exposure and also as controls (CON) for IS and RES. Significant contamination by PAHs was evidenced from both estuarine systems, with higher phenanthrene and chrysene concentrations in the bile of resident fish at BPES, which in turn partially justified the significant induction of EROD and GST in these individuals. Resident fish at BJES showed high EROD and GST induction that cannot be explained by PAHs contamination, and suggests the presence of other contaminants with mechanisms of action similar to dioxins, possibly from a paper industry. Elevation of GST activity was detected in three of the four sites assessed on both estuaries, and loss of swimming resistance was verified on individuals exposed at the same sites, indicating a correlation between GST and this behavioral effect relevant to survival of the species. Indications of acetylcholinesterase inhibitors were not detected, except at the BPES inner region. This study shows the potential and feasibility of using the guppy P. vivipara on the evaluation and monitoring of pollution in estuaries along the Brazilian coast.
MO019
NEW METHODOLOGY TO DETERMINE BTEX IN SOIL SAMPLES BY HPLC-DAD
L. Silva, Universidade Federal do ABC / PROGRAD - CLD; C. da Silva, E.C. Lima, UFABC / CCNH; D. Rosa, UFABC / CECS
Benzene, toluene, ethylbenzene and xylene, commonly referred as BTEX, are constituents of fossil fuels that cause serious negative impacts on the environment and human health. At fuel stations whose storage tanks are leaking, these substances may in contact with the soil and even reach the groundwater. In order to detect the concentration of these compounds in contaminated soils, gas
chromatography (GC) is the most commonly used technique. In the present work it is proposed the use of high performance liquid chromatography coupled to diode array detection (HPLC-DAD) for the determination of BTEX in soil samples. A methodology was developed using as mobile phase methanol and H2O acidified with 250μL of H3PO4 (70:30, v / v), Eclipse XDB C18 column (5μm x 4,6 x 250mm), flow of 1.5 mL min-1, λ = 205nm and T = 50 ° C. The analysis was carried out using the Agilent 1220 HPLC system equipped with an automatic injector, a column oven, and a diode array detector. Data were acquired using the OpenLAB A.01.05 software. A calibration curve for BTEX standards was constructed in 7 concentration levels: 1 to 68 ppm for benzene, 1 to 80 ppm for toluene, 1 to 80 ethylbenzene and 1 to 85 for xylene. The curves were submitted to inter- and intra-assay repeatability analyzes. Standard curves with adjustments above 0.991 relative standard deviations (% RSD) of less than 1.9% were obtained.
Reproducibility tests were performed with two solutions obtained from the standard solution. In the samples containing analytes from the soil contaminated with gasoline, % RSD was obtained below 6.5% and recovery rate was 68% for benzene and 75% for toluene, 78% for ethylbenzene and 78% for xylene. The method of soil analysis via HPLC is therefore efficient and as an alternative to be highlighted for analyzes of soils contaminated with gasoline. It is intended to validate the methodology using appropriate protocols and apply it in contaminated areas for the verification of BTEX levels in a next step.
MO020
Petroleum pollution of alluvial sediments near Sava river, Serbia
M. Ilic, IChTM / Department of Chemistry; S. Bulatovic, Faculty of Chemistry, University of Belgrade; T. Solevic Knudsen, IChTM / Department for Chemistry; J. Milic, Institute of Chemistry, Technology & Metallurgy / Department of Chemistry; S. Miletic, J. Avdalovic, Institute of Chemistry, Technology & Metallurgy, University of Belgrade / Department of Chemistry; G. Devic, Institute of Chemistry, Technology and Metallurgy, University of Belgrade
Heating plant "New Belgrade" is located on the left coast of the Sava River, about 1km from its estuary in the Danube, and represents a potential source of petroleum pollutants for the alluvial area of the river, ground water as well as Sava river. The aim of our research was to determine the presence of petroleum pollutants and their vertical migration in the alluvial area of Sava river. The investigation was started in the summer of 2015. The soil was sampled in three different microlocations (Z1, Z3 and Z7) up to depth of 15m. The sampled material was organized in the layers, and for all microlocations was made a lithological profile. Most of the samples have had a clayey-sand structure with low content of organic matter. Extraction of petroleum pollutants from soil samples were done using the Soxhlet apparatus with dichloromethane. After extraction, the dichloromethane extracts were then fractioned by column chromatography into fractions of: saturated hydrocarbons (Fraction I), aromatic hydrocarbons (Fraction II), and polar compounds (alcohols and keto compounds (Fraction III) [1]. For monitoring changes in the vertical migration of petroleum pollutants, and the relationship of this migration with the soil characteristics, the group composition was determined for each borehole of all
microlocations, taking into account their lithological profiles. Results of our research showed that in all samples the most represented were polar compounds (Fraction III), while the saturated hydrocarbon were least represented (Fraction I). This trend is almost unchanged in samples from different microlocation at different depths. It can be concluded that composition of petroleum pollutants can be unchanged through the alluvial sediments up to 15m depth and they can reach the underground waters, Sava river and consequently disturb the quality of the environment. References: Miletic S., Ilic M., Avdalovic J., Šolevic Knudsen T., Beškoski V.P., Branimir Jovancicevic B., Vrvic M.M. (2015) Oil pollution in the vicinity of a heating plant in New Belgrade (Serbia) – influence on the quality of the surrounding soil and sediments. 16th European Meeting on Environmental Chemistry, EMEC16, Book of Abstracts. November 30 – December 03. 2015, Torino, Italy. Acknowledgements This work was supported in part by Ministry of Education, Science and Technological Development of the Republic of Serbia, Project No: III 43004.
MO021
Prey capture to male aggression: the role of ecologically relevant behaviours in the assessment of complex petroleum based contaminants.
D. Philibert, University of Alberta / Biological Sciences; D. Lyons, C. Philibert, University of Alberta; K.B. Tierney, University of Alberta / Biological Sciences Crude oil and its associated by-products are ubiquitous in the aquatic environment due to both natural and anthropogenic sources (i.e. oil seeps and rivers flowing over surface bitumen, and pipeline ruptures, grounded ships, storage tank leaks and tailing pond seepage, respectively). This diversity in sources gives rise to a large family of complex contaminant mixtures, including weathered and unweathered oil, unconventional oil, such as diluted bitumen (dilbit), and crude oil extraction-based mixtures, such as oil sands process water (OSPW). Historically, studies focused on lethality and cardiotoxicity; complex behaviours have been, for the most part, overlooked despite the merits of including these endpoints in toxicological studies. In this study, we compared various ecologically relevant behaviours (prey capture, male aggression, reaction to alarm odourant) of developmentally exposed fish (Danio rerio and Cyprinodon variegatus variegatus) across various contaminants. Exposure to oil-based contaminants did not impair outright function, but instead altered the variation in behavioral phenotypes present in the population of exposed fishes. Previous studies suggest cortisol can be associated with behavioural phenotypes, and that developmental cortisol levels may pre-determine the behavioural phenotypes found in a population of exposed fishes. Complex behaviours are sensitive sublethal endpoints that could be used in the risk assessment of contaminant mixtures. The inclusion of complex behaviours in toxicological studies brings ecological relevance to a biomarker dominated field.
MO022
Risk-Based Approach: Assessment of Offshore Discharge Waters
K. Wadhia, National Oilwell Varco (NOV) / Environmental; O. Pelz, BP / Gulf Coast Restoration Organization; S. Cousins, BP
In 2012, OSPAR (Oslo and Paris Conventions) adopted the recommendation 2012/5 for a ‘Risk Based Approach (RBA) to the management of Produced Water (PW) discharges from offshore installations’. The application of the RBA recommendations (2012/5) is implemented by the UK’s regulator, the Department for Business, Energy and Industrial Strategy (BEIS). The objective of the RBA is to assess the environmental risk of a PW discharge in the OSPAR maritime area. This is achieved by analysing the effluent and added substances to obtain a measure of the risk of the discharge. If the result is out-with the recommended criteria, a Produced Water Management Plan (PWMP) must be adopted to comply with the RBA regulatory requirements. Processing the information generated by the RBA, each PWMP would be specific to the discharged effluent, platform and area, aiming to minimise environmental risk of each PW discharge. The RBA method is compiled of a six- step process. The steps are based on a standard method where a Predicted Environmental Concentration (PEC) and a Predicted No Effect Concentration (PNEC) of the PW or individual products are determined, and a PEC:PNEC ratio is calculated. The PEC:PNEC ratio and Environmental Impact Factor (EIF) which describes a PEC:PNEC ratio in a specified volume of water characterises the potential risk imposed to the receiving environment. With use of a dispersion modelling tool, the fate of the PW and thus the relative environmental