B R ECOPILACIÓN DE DATOS PARA CERDOS DE ENGORDE EN LA GRANJA ( MEDIDOS EN EL MATADERO )

aquaculture by developing tools, approaches and frameworks to support EU Member States in establishing a coherent and efficient regulatory framework, implementing the Strategic Guidelines for the sustainable development of European aquaculture and delivering a technology and decision framework for sustainable growth. The ultimate goal of the project is to create cost-efficient management tools and practices for the European aquaculture sector to investigate the scope of fish farming activity, social interactions, potential environmental impacts and any future risks.

467

Preliminary investigation on the occurrence of multifunctional organic micropollutants in offshore seawater and fish farm

L. You, National University of Singapore; K. Gin, National University of Singapore / Civil & Environmental Engineering

Limited research has been conducted on the occurrence and distribution of antibiotics, pharmaceuticals, personal care products, endocrine disrupting chemicals and artificial sweeteners in the marine environment despite being increasingly impacted by these micropollutants (MPs). In this study, the presence and distribution patterns of 53 multifunctional organic micropollutants belonging to 14 different groups were investigated in offshore seawaters and fish farms of Singapore. The sampling area is affected by various anthropogenic pressures including treated effluents, fish farming, shipping and port activities. A total of 23 MPs were found in offshore seawaters, 9 of them with detection frequencies higher than 50%. The highest detected values corresponded to cyclamate, salicylic acid and sucralose, with concentration range of

468

Perspectives on Urbanization, Water Reuse, and Aquaculture Product Quality

B.W. Brooks, Baylor University / Dept of Environmental Science; J.L. Conkle, Texas A&M University Corpus Christi / Physical and Environmental Sciencees By 2050, it is estimated that global food production must increase by 50%. Aquaculture will play an important role to meet these needs. For example, in 2014 aquaculture surpassed global fisheries in providing fish for human consumption. It is important to note that global aquaculture activities can in urban and periurban regions with differential waste management capacity. Yet high population densities in urbanizing regions result in concentration of food, energy, water and other resource consumption. Urbanization also leads to concentration of chemical use, which inherently results in exposures to human populations and ecosystems receiving waste streams within and from these urban centers. In developing nations, where many of the megacities will continue to emerge over the next few decades, access to chemical products is occurring faster than public health interventions and environmental management systems are being implemented. Unfortunately, 80% of the global sewage production is not treated, but returned to the environment and thus reused for various purposes. These non-traditional reused waters are being recycled for agriculture, including aquaculture in areas experiencing rapid urbanization, yet implications for water security, food safety and international trade are not routinely examined to manage more sustainable aquaculture practices. In the current presentation we draw from our ongoing efforts in Asia and North America to understand bioaccumulation of organic contaminants of emerging concern in common fish and shellfish used for aquaculture. For example, we have observed aquacultured bivalves to accumulate diverse contaminants of concern (e.g., pharmaceuticals, pesticides, flame retardants), apparently from landfill leachates and effluent discharges of marginal quality, in Hong Kong. Our findings from laboratory uptake and depuration studies with channel catfish and tilapia focus on contaminants with diverse physico-chemical properties (e.g., weak base medicine, phosphorus-based flame retardant, perfluorinated compound, cyanotoxin) and provide an approach to improve aquaculture practice and to support

bioaccumulation assessments for chemicals falling outside of applicability domains for nonionizable organic contaminants. In North America we are examining intersections among water reuse practices and aquaculture for various products. Such efforts apparent warranted at the global scale.

469

Bioaccumulation of selected veterinary medicines in the blue mussel (Mytilus edulis)

S. Brooks, NIVA / Ecotoxicology and Risk Assessment; B. Beylich, NIVA; A. Ruus, NIVA / NIVA; J. Rundberget, NIVA; A. Lillicrap, NIVA / Ecotoxicology and Risk Assessment

Veterinary medicines are widely used within the fish farming industry for the control of sea lice infestation. In 2016, over 10 tonnes of veterinary medicines were used by Norwegian fish farms for the control of sea lice. The impact on non-target species has raised increasing concern. For instance, do wild and farmed mussels in the vicinity to these fish farms have the potential to bioaccumulate these chemicals and thereby pose a threat to human health? On the other hand, mussels may be the most suitable biomonitoring species for the presence of veterinary medicines in the environment. To better understand these scenarios, and the fate of these chemicals in the environment, a series of laboratory controlled exposures were performed to determine the bioaccumulation and depuration of selected veterinary medicines in the blue mussel (Mytilus edulis). The veterinary medicines included teflubenzuron,

emamectin, deltamethrin and azamethiphos. Due to the low solubility of teflubenzuron and deltamethrin a saturation column was employed within a flow-through system to deliver a stable concentration of test chemical over a 14-day uptake phase. Water and mussel samples were collected at time intervals during the 14-day uptake phase, and again following transfer of the mussels into clean flowing seawater during the 7 to 14 day depuration phase. The effects of salinity on the bioaccumulation of teflubenzuron was also investigated to see whether mussels in brackish waters show different bioaccumulation dynamics. So far, the results have shown a clear uptake of teflubenzuron over 14 days, reaching maximum concentrations (~1500 ng/g) after 10 days. Depuration of teflubenzuron was fast for the first 2 days, although still present at approximately 250 ng/g after 7 days depuration. Salinity had no apparent effect on the bioaccumulation of teflubenzuron. In contrast, emamectin showed lower bioaccumulation, with maximum concentrations of 45 ng/g after 6 days. No significant depuration of emamectin was observed after 7 days in clean flowing seawater. The results suggest that mussels are a suitable biomonitoring species for the presence of veterinary medicines in the environment. Additionally, mussel farms in close proximity to fish farms have the potential to bioaccumulate these chemicals in their tissues and is subsequently recommended for monitoring.

470

Contribution of nuclear applications to better understand bioaccumulation of contaminants in aquaculture species

M. Metian, IAEA-EL / Radioecology Lab; S. Pouil, F. Oberhänsli, International Atomic Energy Agency / Environment laboratories; P. Bustamante, Université de La Rochelle / LIttoral ENvironnement et Sociétés LIENSs; P. Swarzenski, International Atomic Energy Agency / Radioecology Lab

Environmental pollution from aquaculture is often seen as a major concern, but today, increasingly is the potential exposure of aquaculture to contaminants. In order to fully understand the contamination risk of farm-raised species, nuclear applications can be used. This is a very powerful approach that allows identifying the susceptibility of economically important species (fish and seafood) to be contaminated. Marine fish farming is regarded as the future of aquaculture and thus, the safety of these farm-raised fish is paramount particularly as 50 % of fish consumed are now farm-raised. Therefore, a better understanding bioaccumulation processes of such contaminants with current aquafarming practices is essential. Such work will attempt to better understand the role the fish food or key

environmental parameters on contamination of fish that may affect the health of the farmed species and/or the human consumer. This has been commonly done in a natural setting but is now beginning to be examined for fish farming practices. Major advantages of radiotracer techniques over conventional techniques are their very high sensitivity and discrimination capacity: it permits the measurement of bioaccumulation kinetics of several elements at realistic (viz. low) environmental concentrations in a single experiment. Furthermore, some radiotracer permits the non-destructive analyses of contaminant levels in living organisms. This paper identifies present and future threats on farm-raised fish from a contamination point of view, and presents a synthesis of experimental results completed on farm-raised fish exposed to contaminants in realistic environmental conditions. It revealed, for example, the various effects that food, water salinity and temperature can have on the Assimilation efficiencies of trace elements and radionuclides in farmed fish. Such findings paving the way for further investigations on the potential use of nuclear techniques in aquaculture and food safety.

471

Effects of antibiotic´s medicated fish feed in the marine environment

B. Gonzalez-Gaya, IMDEA Water (G84912732) / Environmental Chemistry; N. García Bueno, I. Gomez, B. Martinez-Lopez, P. Franco, University of Murcia / Ecology and Hydrology; E. Buelow, Limoges University / Medicine Faculty, Inserm Umr 1092; A. Marin, University of Murcia / Ecology and Hydrology; A. Rico, IMDEA Water Institute / Aquatic Ecotoxicology

Intensive aquaculture is considered to be an important source of antibiotics into the marine environment. Antibiotics used in aquaculture have been reported to accumulate on sediments and non target aquatic organisms, modifying the biodiversity and the environmental conditions in areas close to fish farms. Moreover, recent studies show an increasing ocurrence of resistance genes in environmental bacteria next to fish farms, which indicates the assembly, selection and dissemination of antimicrobial resistance through open aquaculture installations and might thereby compromise environnmental and human health. A field experiment was performed in a moderatelly impacted bay in the south east coast of Spain (Aguilas, Murcia), which consisted of a series of sediment traps (covered by a net vs. uncovered) filled with local sediment and fish feed; non medicated or medicated with three antibiotics (oxytetracycline, florfenicol and flumequine). Fish feeds were applied simulating fish farm loses for a period of 3 weeks. Measured antibiotic concentrations in the sediment were 2700 – 8000 ng/g (average 1% of the applied ammount) for oxytetracycline, and 19000 – 54000 ng/g (average 10% of applied amount) for flumequine. Florfenicol was not detected. Different accumulation rates were found in covered/uncovered traps due to wild fish influences in the availability of feed and bioturbation. Physico-chemical characteristics of the sediment also changed; with a higher S and lower N content and a larger percentage of fine material in feed affected treatments. Invertebrate

presence was also correlated with the food availability, although no evident effects of the antibiotics were found over the analyzed samples. Bioaccumulation of the target antibiotics in the invertebrate community and evaluation of the antibiotic impacts over the microbiome and resistome of the sediment bacteria is still ongoing. This is one of the first studies describing fish feed and antibiotic impacts produced by aquaculture under Mediterranean conditions.

Systems ecotoxicology: application of OMICS data across

multiple level of biological organization in research and risk

assessment (II)

472

Systems toxicology approach for the assessment of zebrafish cardiac and neurotoxicity

R. Li, M. Talikka, Philip Morris International; S. Madan, Fraunhofer Institute for Algorithms and Scientific Computing; J. Doerpinghaus, Fraunhofer Institute for Algorithms and Scientific Computing; A. Zupanic, Eawag Swiss Federal Institute of Aquatic Science and Technology / Environmental Toxicology; J. Fluck, Fraunhofer Institute for Algorithms and Scientific Computing; C.M. vom Berg, Eawag Swiss Federal Institute of Aquatic Science and Technology / Environmental Toxicology; J. Szostak, F. Martin, M. Peitsch, J. Hoeng, Philip Morris International A major goal in the field of toxicology is to predict long term animal health risks and/or environmental hazards associated with a particular substance(s).

Traditionally utilised classical toxicology methods involve animal exposure over a relatively short period and recording adverse outcomes. These data are then extrapolated to long term effects and to other species. The accuracy of such extrapolation would benefit from mechanistic understanding of toxicity. However, molecular basis for adverse outcomes is not easily interpreted from classical toxicology methods. Here we present our systems toxicology approach that focuses on deciphering biological mechanisms responsible for adverse outcomes. The underlying structure of this approach is a computable biological network model. We have developed two models describing molecular pathways that lead to cardiotoxicity and neurotoxicity in zebrafish larvae based on the knowledge curated from scientific literature. Key signalling nodes in the model are linked to information about downstream gene expression. Differential expression of downstream genes can be used to infer activity of the upstream protein – a process termed network scoring. Scoring of the network highlights the most affected nodes, which leads to mechanistic hypothesis generation and gives a quantifiable measure of network perturbation. In parallel to network scoring, we utilize classical toxicology methods to detect adverse outcomes. We present the acute toxicity results for selected chemicals (e.g. acrylamide, arsenic, citalopram, imidacloprid) according to the OECD fish embryo acute toxicity test (OECD test guideline 236). We then report results from chemically exposed larvae in functional cardiac and behavioural assays, and transcriptomics analyses. Finally, we describe the utility of the network model in interpreting transcriptomics analyses to gain mechanistic insight into the molecular events initiated by a given chemical. Cardiac and neural apical endpoints together with computational network scoring provide a comprehensive method for linking molecular events to organ toxicity. This approach will enable more accurate toxicity predictions over long exposures and in different species.

473

Time response relationship between gene expression and life history in a Daphnia population exposed to heavy metals

J. Asselman, I. Semmouri, Ghent University / Laboratory for Environmental Toxicology and Aquatic Ecology GhEnToxLab unit; K. De Schamphelaere, Ghent University (UGent) / Applied Ecology and Environmental Biology

Over the last decade, molecular technologies have evolved into robust high throughput platforms available to many scientists in a wide variety of disciplines. Implementation of these technologies in ecotoxicology and risk assessments have focused on mechanisms of toxicity and stress response on the gene level to explain effects at the organism level. However, current studies remain focused at the individual level and rarely include population level molecular responses. Population level molecular responses may provide a better insight into the potential mechanisms at play at the population level while at the same time avoid focusing on gene expression patterns that are the cause of clonal or interindividual variation. Furthermore, most studies select an arbitrary timepoint to measure gene expression responses without any prior knowledge. Here, we focus on population level responses of a Daphnia magna population to arsenic and copper and their binary mixture. The population was exposed to low chronic toxicity concentrations of arsenic and copper resulting primarily in effects on reproduction rather than survival. Rather than focusing on a single arbitrary timepoint, gene expression data and life history data were both recorded at multiple time points. As such, these datasets will provide a first basis on how exposure duration may affect the conclusions and decisions made about the toxicity of chemicals. In addition, by collecting both molecular data and life history data, we will be able to better understand the time response relationship in populations under stress both at the life history level and the molecular level. This will allow us to better integrate these two data types and identify potential causal relationships between the molecular level

and the life history level. The identification of such causal relationships will play an integral part of incorporating omics data in environmental risk assessment.

474

How to implement functional responses of microalgae in risk assessment processing?

F. Larras, Helmholtz Center for Environmental Research - UFZ GmbH; E. Billoir, Université de Lorraine, CNRS UMR 7360; S. Scholz, Helmholtz Centre for Environmental Research / Department Bioanalytical Ecotoxicology; M. Delignette-Muller, VetAgro Sup / Laboratory of Biometry and Evolutionnary Biology; M. Schmitt-Jansen, UFZ - Helmholz Ctre Environm. Research / Department of Bioanalytical Ecotoxicology

Microorganisms (e.g. bacteria, fungi and algae) are involved in various ecosystem functions such as biogeochemical cycles or pollutants degradation meaning that they are crucial for ecosystem functioning. In the environment, organisms are exposed to anthropogenic pressures which are known to potentially induce structural and functional changes. If such causal links are identified, little is known about the involved biochemical pathways supporting specific functions. Moreover, most of a priori ecological risk assessment (ERA) tools are based on structural endpoints and do not necessarily ensure the protection of these functions. The recent raise of OMICs approaches (e.g. transcriptomics and metabolomics) opens the perspective in ecotoxicology to explore pathways involved in ecological functions. The main aim of this study was to provide a new and innovative risk assessment tool based on functional responses (captured via OMICs approaches) of periphytic communities in order to protect the functions that they ensured. The rationale to use OMICs in such context is to provide more protective and early warning thresholds. The transcriptomic and the metabolomic responses of

Scenedesmus vacuolatus to triclosan were explored after exposure of 14 hours

along an increasing gradient of 5 concentrations (from 0.69 to 6.63 µg/L, 5 replicates). Within a dedicated workflow, we selected the responsive molecular items (metabolites/transcripts), we built concentration response curves for each of them and we derived a sensitivity value from each curve (even the non-monotonic one). Molecular items showed mainly non-sigmoid and even non-monotonic responses to triclosan exposure. For example, the transcripts data were mainly best described by an exponential model for more than half of the curves and a Gaussian or log-Gaussian model for more than a quarter of the curves. Then, the molecular items were linked (when possible) to the pathways they are involved in. From that information, we built a SSD-like tool based on functional responses captured at the community level in order to protect functions and integrating two levels of OMICs responses. The next step consists to build such tool from the periphytic community level.

475

Sex, drugs and Daphnia magna. A multi-omics approach suggests conserved mechanisms of interaction between metallohormones and endocrine disruptors

E. Caamano-Gutierrez, University of Liverpool / Computational Biology Facility; P. Antczak, University of Liverpool / Institute of Integrative Biology; L. Mirbahai, The University of Birmingham / School of Biosciences; K. Grintzalis, The University of Birmingham; M.R. Viant, University of Birmingham / School of Biosciences; F. Falciani, University of Liverpool / Institute of Integrative Biology The assessment of environmental exposure to toxic chemicals released by human activity as well as their impact on biological systems is key to protect the biosphere. Current environmental monitoring protocols are based on chemical analysis and an assessment of biodiversity. Although this has been a very effective strategy, it has some shortcomings. These include the fact that a relatively limited number of compounds can be measured and linked to biologically relevant organism-level responses. The issue is particularly challenging in chronic exposures and in complex mixtures scenarios. This project aims at identifying the molecular networks linked to single and mixture exposures and to use these to infer the effects of chemical mixtures. We approached this important challenge by applying a systems biology approach to integrate expression profiling, metabolomics and phenotypic data (respiration and feeding rates), representing the response of

Daphnia magna to a panel of environmentally relevant chemicals and their

mixtures. Firstly, it was exposed to a battery of single compounds with known mode of action (MoA) i.e. estrogen disruptors and acetylcholine esterase (AChE) inhibitors as well as metals with unclassified MoA. We have been able to model the differences between the two main MoA studied and linked them to biological activities within Daphnia. Furthermore, we have found that as expected, metals do not show a common MoA, with some of them clustering closer to either endocrine disruptors or AChE inhibitors. Cadmium (Cd), which clustered with endocrine disruptors, has already been shown to play a role with the estrogen receptor in humans but its role in D. magna is still under surveillance. To further study this finding we exposed D. magna to complex mixtures of Cd and ethinylestradiol. While the individual exposures triggered the alteration of expression of a relatively large number of genes, the exposure to the mixture showed little or no effect. These results indicated that both compounds share a complex interaction at a molecular level suggesting that the degree of conservation of the regulatory pathways