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Despite the importance of bacteria in surf zone water quality, detailed insights into their community composition, functions, and seasonal dynamics at recreational beaches are scarce. This study conducted year-long, weekly monitoring of bacterial communities and environmental factors at a recreational beach in Ostend, Belgium. Using full-length 16S rRNA gene sequencing, we correlated bacterial composition and predicted functions with environmental factors to identify potential drivers. Bacterial communities were significantly affected by seasonal variations in chlorophyll a (Chl a), net primary productivity (NPP), and seawater temperature (SWT), with minimal influence from faecal inputs due to human activities. Spring showed distinct abundances of Planktomarina, Amylibacter, and Sulfitobacter, positively correlated with Chl a and related to sulphur oxidation potential. Summer had higher abundances of Cryomorphaceae, likely enhancing chemoheterotrophy. Beginning in mid to late fall and extending into winter, bacterial communities underwent substantial changes. Fall featured a distinctive enrichment of Thioglobaceae, inversely correlated with Chl a. Winter was dominated by Methylophilaceae (OM43 clade), negatively correlated with Chl a, NPP, and SWT. Both seasons exhibited elevated levels of potentially pathogenic phenotypes and predicted functions related to methanol oxidation and methylotrophy. This study provides a baseline for understanding how surf zone bacterial communities respond to environmental changes and impact health., (© 2024 The Author(s). Environmental Microbiology Reports published by John Wiley & Sons Ltd.)

Consuming the meat of some marine turtles can lead to a specific type of seafood poisoning known as chelonitoxism. A recent poisoning event (March 2024) on the Tanzanian island Pemba, resulting in the death of 9 people and hospitalization of 78 others, underscores the need to obtain an up to date overview and understanding of chelonitoxism. Here, we document a global overview of poisoning incidents resulting from the consumption of sea turtle flesh worldwide. All events combined involved over 2400 victims and 420 fatalities. Incidents were predominantly reported in remote regions (often islands) across the Indo-Pacific region. Reported health effects of consuming poisonous sea turtles include epigastric pain, diarrhea, vomiting, a burning mouth and throat sensation, and dehydration. In addition, ulcerative oeso-gastro-duodenal lesions, which occasionally have resulted in hospitalization and death, have been reported. Lyngbyatoxins have been suggested as (one of) the causative agents, originating from the cyanobacterium Moorena producens, growing epiphytically on the seagrass and seaweed consumed by green turtles. However, due to the limited evidence of their involvement, the actual etiology of chelonitoxism remains unresolved and other compounds may be responsible. The data outlined in this review offer valuable insights to both regulatory bodies and the general public regarding the potential risks linked to consuming sea turtles., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Bivalve hemocytes, particularly granulocytes and hyalinocytes, play a crucial role in cell-mediated immunity. However, their interactions with aged plastic particles, exhibiting altered properties that more closely resemble those in natural environments, remain largely underexplored. This study assesses the differential responses of hemocyte subpopulations (Mytilus edulis) to chemically aged polyethylene terephthalate (PET) microplastic (MPs) and nanoplastic (NPs) particles across multiple cellular effect endpoints. Particle characteristics were analyzed using Single Particle Extinction and Scattering, Raman Spectroscopy, Scanning Electron Microscopy, and Dynamic Light Scattering. In vitro experiments with aged PET MPs (1.9 µm) and NPs (0.68 µm) were conducted at three internally relevant concentrations: 10 (C1), 10³ (C2), and 10⁵ particles/mL (C3). Cellular responses were assessed by measuring lysosomal content stability, reactive oxygen species (ROS) production, cellular mortality, and morphological parameters using flow cytometry at 6, 12, 24, and 48 hours. Our findings provide mechanistic insights into the differential sensitivities of granulocytes and hyalinocytes to aged PET, influenced by particle size and concentration. Specifically, aged PET MPs and NPs induce distinct size and concentration-dependent patterns of lysosomal destabilization, coinciding with the loss of functional integrity. Elevated ROS levels were observed only in granulocytes and hyalinocytes exposed to high concentrations of aged PET NPs, underscoring the effects on oxidative stress. Both aged PET MPs and NPs induce significant increases in cellular mortality, particularly after 24 h of exposure at high concentrations. These findings reveal the complex cellular mechanisms underlying hemocyte functional impairment following exposure to aged PET particles under environmentally and biologically relevant conditions., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Raewyn M. Town reports a relationship with Ecotoxicology and Environmental Safety that includes: board membership. Co-author is a member of the International Editorial Board of Ecotoxicology and Environmental Safety - R.M.T. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Despite the urgent need for accurate and robust observations of microplastics in the marine environment to assess current and future environmental risks, existing procedures remain labour-intensive, especially for smaller-sized microplastics. In addition to this, microplastic analysis faces challenges due to environmental weathering, impacting the reliability of research relying on pristine plastics. This study addresses these knowledge gaps by testing the robustness of two automated analysis techniques which combine machine learning algorithms with fluorescent colouration of Nile red (NR)-stained particles. Heterogeneously shaped uncoloured microplastics of various polymers-polyethylene (PE), polyethylene terephthalate (PET), polypropylene (PP), polystyrene (PS), and polyvinyl chloride (PVC)-ranging from 100 to 1000 µm in size and weathered under semi-controlled surface and deep-sea conditions, were stained with NR and imaged using fluorescence stereomicroscopy. This study assessed and compared the accuracy of decision tree (DT) and random forest (RF) models in detecting and identifying these weathered plastics. Additionally, their analysis time and model complexity were evaluated, as well as the lower size limit (2-4 µm) and the interoperability of the approach. Decision tree and RF models were comparably accurate in detecting and identifying pristine plastic polymers (both > 90%). For the detection of weathered microplastics, both yielded sufficiently high accuracies (> 77%), although only RF models were reliable for polymer identification (> 70%), except for PET particles. The RF models showed an accuracy > 90% for particle predictions based on 12-30 pixels, which translated to microplastics sized < 10 µm. Although the RF classifier did not produce consistent results across different labs, the inherent flexibility of the method allows for its swift adaptation and optimisation, ensuring the possibility to fine-tune the method to specific research goals through customised datasets, thereby strengthening its robustness. The developed method is particularly relevant due to its ability to accurately analyse microplastics weathered under various marine conditions, as well as ecotoxicologically relevant microplastic sizes, making it highly applicable to real-world environmental samples., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Harmful algal blooms (HABs) - proliferated algae densities, often producing toxins - have increasingly been found in ocean and coastal areas. Recent studies show that rising temperatures contribute to HAB occurrence, but the broader influence of climate change on these outbreaks is less quantified. Of particular concern is the limited research on HAB toxin effects under varying temperatures, especially regarding primary consumers such as copepods, a crucial component of aquatic ecosystems. Therefore, we examined the impact of marine toxins on the harpacticoid copepod Nitokra spinipes, a model organism for marine ecotoxicology, in the context of climate change. We evaluated the toxicity of four purified, commonly occurring algal toxins, at three different temperatures in the laboratory. First, adult females were exposed to a concentration series of toxins at 15, 20, and 25 °C for 48 h. EC50 values of domoic acid ranged from 8.79 ± 1.93 μg L -1 to 25.97 ± 11.96 μg L -1 . Nauplii, aged 48-72 h, were exposed at 18, 20 and 22 °C for the same duration. Less sensitive compared to adults, the EC50 of domoic acid in this case varied from 57.26 ± 6.82 μg L -1 to 97.24 ± 6.45 μg L -1 . Both results indicated a temperature-dependent EC50. For the chronic toxicity tests, larval development ratio (LDR), brood size and inter-brood time of domoic acid (DA), yessotoxin (YTX), saxitoxin (STX), and microcystin-LR (MC-LR) were examined at 18, 20 and 22 °C. We observed that with increasing temperatures, LDR increased, whereas brood size significantly decreased as DA, YTX or STX concentrations rose. No interaction between temperature and algal toxins was found but a temperature dependent sensitivity of copepods towards DA, YTX and STX was revealed. Our research provides insights into the effects of long-term exposure to algal toxins on marine copepods and the potential impacts of climate warming., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Microplastic (MP) research faces challenges due to costly, time-consuming, and error-prone analysis techniques. Additionally, the variability in data quality across studies limits their comparability. This study addresses the critical need for reliable and cost-effective MP analysis methods through validation of a semi-automated workflow, where environmentally relevant MP were spiked into and recovered from marine fish gastrointestinal tracts (GITs) and blue mussel tissue, using Nile red staining and machine learning automated analysis of different polymers. Parameters validated include trueness, precision, uncertainty, limit of quantification, specificity, sensitivity, selectivity, and method robustness. For fish GITs a 95 ± 9 % recovery rate was achieved, and 87 ± 11 % for mussels. Polymer identification accuracies were 76 ± 8 % for fish GITs and 80 ± 13 % for mussels. Polyethylene terephthalate fragments showed more variability with lower accuracies. The proposed validation parameters offer a step towards quality management guidelines, as such aiding future researchers and fostering cross-study comparability., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Micro- and nanoplastics (MNPs) can enter the atmosphere via sea spray aerosols (SSAs), but the effects of plastic characteristics on the aerosolization process are unclear. Furthermore, the importance of the transport of MNPs via these SSAs as a possible new exposure route for human health remains unknown. The aim of this study was two-fold: (1) to examine if a selection of factors affects aerosolization processes of MNPs, and (2) to estimate human exposure to MNPs via aerosols inhalation. A laboratory-based bubble bursting mechanism, simulating the aerosolization process at sea, was used to investigate the influence of MNP as well as seawater characteristics. To determine the potential human exposure to microplastics via inhalation of SSAs, the results of the laboratory experiments were extrapolated to the field based on sea surface microplastic concentrations and the volume of inhaled aerosols. Enrichment seemed to be influenced by MNP size, concentration and polymer type. With higher enrichment for smaller particles and denser polymers. Experiments with different concentrations showed a larger range of variability but nonetheless lower concentrations seemed to result in higher enrichment, presumably due to lower aggregation. In addition to the MNP characteristics, the type of seawater used seemed to influence the aerosolization process. Our human exposure estimate to microplastic via inhalation of sea spray aerosols shows that in comparison with reported inhaled concentrations in urban and indoor environments, this exposure route seems negligible for microplastics. Following the business-as-usual scenario on plastic production, the daily plastic inhalation in coastal areas in 2100 is estimated to increase but remain far below 1 particle per day. This study shows that aerosolization of MNPs is a new plastic transport pathway to be considered, but in terms of human exposure it seems negligible compared to other more important sources of MNPs, based on current reported environmental concentrations., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The impact of multiple environmental and anthropogenic stressors on the marine environment remains poorly understood. Therefore, we studied the contribution of environmental variables to the densities and gene expression of the dominant zooplankton species in the Belgian part of the North Sea, the calanoid copepod Temora longicornis. We observed a reduced density of copepods, which were also smaller in size, in samples taken from nearshore locations when compared to those obtained from offshore stations. To assess the factors influencing the population dynamics of this species, we applied generalised additive models. These models allowed us to quantify the relative contribution of temperature, nutrient levels, salinity, turbidity, concentrations of photosynthetic pigments, as well as chemical pollutants such as polychlorinated biphenyls and polycyclic aromatic hydrocarbons (PAHs), on copepod density. Temperature and Secchi depth, a proxy for turbidity, were the most important environmental variables predicting the densities of T. longicornis, followed by summed PAH and chlorophyll concentrations. Analysing gene expression in field-collected adults, we observed significant variation in metabolic and stress-response genes. Temperature correlated significantly with genes involved in proteolytic activities, and encoding heat shock proteins. Yet, concentrations of anthropogenic chemicals did not induce significant differences in the gene expression of genes involved in the copepod's fatty acid metabolism or well-known stress-related genes, such as glutathione transferases or cytochrome P450. Our study highlights the potential of gene expression biomonitoring and underscores the significance of a changing environment in future studies., (© 2024 John Wiley & Sons Ltd.)

To better understand the fate and assess the ingestible fraction of microplastics (by aquatic organisms), it is essential to quantify and characterize of their released from larger items under environmental realistic conditions. However, the current information on the fragmentation and size-based characteristics of released microplastics, for example from bio-based thermoplastics, is largely unknown. The goal of our work was to assess the fragmentation and release of microplastics, under ultraviolet (UV) radiation and in seawater, from polylactic acid (PLA) items, a bio-based polymer, and from polypropylene (PP) items, a petroleum-based polymer. To do so, we exposed pristine items of PLA and PP, immersed in filtered natural seawater, to accelerated UV radiation for 57 and 76 days, simulating 18 and 24 months of mean natural solar irradiance in Europe. Our results indicated that 76-day UV radiation induced the fragmentation of parent plastic items and the microplastics (50 - 5000 µm) formation from both PP and PLA items. The PP samples (48 ± 26 microplastics / cm 2 ) released up to nine times more microplastics than PLA samples (5 ± 2 microplastics / cm 2 ) after a 76-day UV exposure, implying that the PLA tested items had a lower fragmentation rate than PP. The particles' length of released microplastics was parameterized using a power law exponent (α), to assess their size distribution. The obtained α values were 3.04 ± 0.11 and 2.54 ± 0.06 (-) for 76-day UV weathered PP and PLA, respectively, meaning that PLA microplastics had a larger sized microplastics fraction than PP particles. With respect to their two-dimensional shape, PLA microplastics also had lower width-to-length ratio (0.51 ± 0.17) and greater fiber-shaped fractions (16%) than PP microplastics (0.57 ± 0.17% and 11%, respectively). Overall, the bio-based PLA items under study were more resistant to fragmentation and release of microplastics than the petroleum-based PP tested items, and the parameterized characteristics of released microplastics were polymer-dependent. Our work indicates that even though bio-based plastics may have a slower release of fragmented particles under UV radiation compared to conventional polymer types, they still have the potential to act as a source of microplastics in the marine environment, with particles being available to biota within ingestible size fractions, if not removed before major fragmentation processes., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: The authors Elke Demeyer is employed by Centexbel (Ghent, BE), a private company highly active in the knowledge transfer of textile and plastic processing industry. The other authors report no conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Understanding the sinking behavior of microplastics in freshwater is essential for assessing their environmental impact, guiding research efforts, and formulating effective policies to mitigate plastic pollution. Sinking behavior is a complex process driven by plastic density, environmental factors and particle characteristics. Moreover, the growth of biological entities on the plastic surface can affect the total density of the microplastics and thus influence the sinking behavior. Yet, our understanding of these processes in freshwater is still limited. Our research thus focused on studying biofilm growth on microplastics in freshwater. Therefore, we evaluated biofilm growth on five different polymer types (both microplastic particles and plates) which were incubated in freshwater for 63 days in a controlled laboratory setting. Biofilm growth (mass-based) was used to compare biofilm growth between polymer types, surface roughness and study the changes over time. Understanding the temporal aspect of biofilm growth enabled us to refine calculations on the predicted effect of biofilm growth on the settling behavior in freshwater. The results showed that biofilm formation is polymer-specific but also affected by surface roughness, with a rougher surface promoting biofilm growth. For PET and PS, biofilm tended to grow exponentially during 63 days of incubation. Based on our calculations, biofilm growth did affect the sinking behavior differently based on the polymer type, size and density. Rivers can function as sinks for some particles such as large PET particles. Nevertheless, for others, the likelihood of settling within river systems appears limited, thereby increasing the probability of their transit to estuarine or oceanic environments under hydrometeorological influences. While the complexity of biofilm dynamics on plastic surfaces is not fully understood, our findings help to elucidate the effect of biofilms on the vertical behavior of microplastics in freshwater systems hereby offering knowledge to interpret observed patterns in environmental plastic concentrations., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Jana Asselman reports financial support was provided by Flanders Innovation & Entrepreneurship., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Plastics are omnipresent in our daily life. Unfortunately, the produced plastics will partly end up in the environment including aquatic ecosystems. People often refer to littering or illegal waste dumping as sources of plastic emission to the environment. However, daily-life sources could also, unknowingly, contribute considerably to the total microplastic pollution in the ecosystem. Hence, there is an urgent need to study these potential sources. In this research, two common sources, i.e. domestic wastewater and road run-off from tire and road wear particles, were studied in detail to quantify the relative contribution of both domestic sources towards microplastic pollution in freshwater ecosystems in Flanders, Belgium. This assessment shows that every person (in studied area) emits on average 1145 microplastics (25-1000 μm) daily through domestic wastewater, resulting in a yearly discharge of 418,000 microplastic particles per person. The road run-off samples contained between 0.02 and 9.2 mg tire wear particles per litre per day, which corresponds to an emission of 10.8 mg tire wear particles per driven vehicle km. The gross and net emissions of both above mentioned microplastic sources were extrapolated to the whole Flanders region using an emission model. From the yearly gross microplastic pollution in the domestic wastewater, 623 kg (20%) will be discharged in the freshwater. The highest losses originated from the households that have a private drain or are not (yet) connected to an active wastewater treatment plant. In Flanders, the yearly net microplastic emission into the aquatic environment of tire wear particles is estimated to be 246 tonnes (38%), mainly from the direct run-off from the road surface. Based on the results, specific mitigation measures can be installed to reduce the emission of microplastics towards the freshwater ecosystem. Other sources should be quantified in a similar way for a more holistic strategy to counteract plastic pollution., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Colin R. Janssen reports financial support was provided by EMBRC Belgium - FWO international research infrastructure. Colin R. Janssen reports financial support was provided by Flemish government (Toegepast Wetenschappelijk Onderzoek Leefmilieu, TWOL)., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Time-series are crucial to understand the status of zooplankton communities and to anticipate changes that might affect the entire food web. Long-term time series allow us to understand impacts of multiple environmental and anthropogenic stressors, such as chemical pollution and ocean warming, on the marine ecosystems. Here, a recent time series (2018-2022) of abundance data of four dominant calanoid and one harpacticoid copepod species from the Belgian Part of the North Sea was combined with previously collected (2009-2010, 2015-2016) datasets for the same study area. The time series reveals a significant decrease (up to two orders of magnitude) in calanoid copepod abundance (Temora longicornis, Acartia clausi, Centropages spp., Calanus helgolandicus), while this was not the case for the harpacticoid Euterpina acutifrons. We applied generalized additive models to quantify the relative contribution of temperature, nutrients, salinity, primary production, turbidity and pollution (anthropogenic chemicals, i.e., polychlorinated biphenyls and polycyclic aromatic hydrocarbons) to the population dynamics of these species. Temperature, turbidity and chlorophyll a concentrations were the only variables consistently showing a relative high contribution in all models predicting the abundances of the selected species. The observed heat waves which occurred during the summer periods of the investigated years coincided with population collapses (versus population densities in non-heatwave years) and are considered the most likely cause for the observed copepod abundance decreases. Moreover, the recorded water temperatures during these heatwaves correspond to the physiological thermal limit of some of the studied species. As far as we know, this is the first study to observe ocean warming and marine heat waves having such a dramatic impact (population collapse) on the dominant zooplankton species in shallow coastal areas., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Sea spray has been suggested to enable the transfer of micro- and nanoplastics (MNPs) from the ocean to the atmosphere, but only a few studies support the role of sea spray aerosols (SSAs) as a source of airborne particles. We demonstrated that MNPs are aerosolized during wave action, via SSAs, under controlled laboratory conditions. We used a mini-Marine-Aerosol-Reference-Tank (miniMART), a device that mimics naturally occurring physical mechanisms producing SSAs, and assessed the aerosolization of fluorescent polystyrene beads (0.5-10 μm), in artificial seawater. The SSAs contained up to 18,809 particles/mL of aerosols for 0.5 μm beads, with an enrichment factor of 19-fold, and 1977 particles/mL of aerosols for 10 μm beads with a 2-fold enrichment factor. Our study demonstrates that the use of the miniMART is essential to assess MNPs aerosolization in a standardized way, supporting the hypothesis which states that MNPs in the surface of the ocean may be transferred to the atmosphere., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Microplastics (1 µm-5 mm), are ubiquitous in daily-use products and regularly end up in the wastewater. The main part of the wastewater is treated in wastewater treatment plants (WWTPs), which allow for at least partial removal of microplastics. The present study aimed to understand the contribution of domestic wastewater to microplastic pollution in Flanders (Belgium) via two main discharge routes of microplastics: (1) the effluent, and (2) removed fractions. Furthermore the effect of effluent discharge on the microplastic contamination in the waterway was studied in both surface water and sediment samples of upstream and downstream locations of the discharge from three WWTPs. On average, 12.64 ± 20.20 microplastic/L entered a WWTP (10 µm-5 mm). The effluent contained on average 0.41 ± 0.91 microplastic/L, resulting in an average removal efficiency of 97.46% ± 2.33%, which is comparable with various (non-)European countries. Removal efficiencies are both polymer- and size-specific, and data suggest that smaller particles are less efficiently removed from the wastewater, which also causes an increased input of smaller particles to the environment. The sludge is the most efficient treatment process to remove microplastics. Despite the high removal efficiencies, still 1.11 × 10 7  ± 3.07 × 10 7 microplastics end up in the nearby waterway daily. Nonetheless, based on the results gathered in the present study, this does not seem to impact the microplastic concentration in the waterway significantly. In summary, the present study offers a holistic approach in the research on the impact of wastewater on microplastic pollution in the ecosystem, integrating different discharge routes and measuring the impact on environmental microplastic pollution. Environ Toxicol Chem 2023;42:642-654. © 2022 SETAC., (© 2022 SETAC.)

Marine environments are globally impacted by vast quantities of munition disposed following both World Wars. Dumped munitions contain conventional explosives, chemicals warfare agents as well as a variety of metals. Field monitoring studies around marine dumpsites report the presence of munition constituents in water and sediment samples. The growing interest and developments in the ocean as a new economic frontier underline the need to remediate existing dumpsites. Here, we provide a comprehensive assessment of the magnitude and potential risks associated with marine munition dumpsites. An overview of the global distribution of dumpsites identifying the most impacted areas is provided, followed by the currently available data on the detection of munition constituents in environmental samples and evidence of their toxic potential to human and environmental health. Finally, existing data gaps are identified and future research needs promoting better understanding of the impact of the dumped material on the marine environment suggested., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Microplastic is ubiquitously and persistently present in the marine environment, but knowledge of its population-level effects is limited. In the present study, to quantify the potential theoretical population effect of microplastic, a two-step approach was followed. First, the impact of microplastic (polyethylene, 0.995 g cm -3 , diameter 10-45 µm) on the filtration rate of the pelagic copepod Temora longicornis was investigated under laboratory conditions. It was found that the filtration rate decreased at increasing microplastic concentrations and followed a concentration-response relationship but that at microplastic concentrations <100 particles L -1 the filtration rate was not affected. From the concentration-response relationship between the microplastic concentrations and the individual filtration rate a median effect concentration of the individual filtration rate (48 h) of 1956 ± 311 particles L -1 was found. In a second step, the dynamics of a T. longicornis population were simulated for realistic environmental conditions, and the effects of microplastics on the population density equilibrium were assessed. The empirical filtration rate data were incorporated in an individual-based model implementation of the dynamic energy budget theory to deduct potential theoretical population-level effects. The yearly averaged concentration at which the population equilibrium density would decrease by 50% was 593 ± 376 particles L -1 . The theoretical effect concentrations at the population level were 4-fold lower than effect concentrations at the individual level. However, the theoretical effect concentrations at the population level remain 3-5 orders of magnitude higher than ambient microplastic concentrations. Because the present experiment was short-term laboratory-based and the results were only indirectly validated with field data, the in situ implications of microplastic pollution for the dynamics of zooplankton field populations remain to be further investigated. Environ Toxicol Chem 2022;41:1663-1674. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC., (© 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.)

Indole signaling plays an important role in bacterial pathogenesis. In this study, the impact of indole on biofilm formation, swimming and swarming motility were explored in Vibrio tasmaniensis LGP32 and Vibrio crassostreae J2-9, two model pathogens of bivalves. The results showed that indole decreased swimming and swarming motility in both strains, and decreased biofilm formation in V. crassostreae J2-9. Furthermore, indole affected a large number of genes at RNA level, including genes related to metabolism, ABC transporters, flagellar assembly, chemotaxis, and response regulators. Finally, the bacterial virulence towards mussel larvae was decreased by pretreatment with indole in both V. tasmaniensis LGP32 and V. crassostreae J2-9. After 5 days, the survival rate of mussel larvae increased 2.4-fold and 2.8-fold in mussel larvae challenged with V. tasmaniensis LGP32 pretreated with 200 µM and 500 µM indole, respectively. The survival rate of mussel larvae increased 1.5-fold and 1.9-fold in mussel larvae challenged with V. crassostreae J2-9 pretreated with 200 µM and 500 µM indole, respectively. These data indicate that indole has a significant impact on the virulence of V. tasmaniensis LGP32 and V. crassostreae J2-9, and indole signaling could be a promising target for antivirulence therapy., (© 2022. The Author(s).)

Marine phytoplankton influence the composition of sea spray aerosols (SSAs) by releasing various compounds. The biogenic surfactant dipalmitoylphosphatidylcholine (DPPC) is known to accumulate in the sea surface microlayer, but its aerosolization has never been confirmed. We conducted a 1 year SSA sampling campaign at the Belgian coast and analyzed the SSA composition. We quantified DPPC at a median and maximum air concentration of 7.1 and 33 pg m -3 , respectively. This discovery may be of great importance for the field linking ocean processes to human health as DPPC is the major component of human lung surfactant and is used as excipient in medical aerosol therapy. The natural airborne exposure to DPPC seems too low to induce direct human health effects but may facilitate the effects of other marine bioactive compounds. By analyzing various environmental variables in relation to the DPPC air concentration, using a generalized linear model, we established that wave height is a key environmental predictor and that it has an inverse relationship. We also demonstrated that DPPC content in SSAs is positively correlated with enriched aerosolization of Mg 2+ and Ca 2+ . In conclusion, our findings are not only important from a human health perspective but they also advance our understanding of the production and composition of SSAs.

To date, few studies have examined the role of sea spray aerosols (SSAs) in human exposure to harmful and beneficial marine compounds. Two groups of phycotoxins (brevetoxins and ovatoxins) have been reported to induce respiratory syndromes during harmful algal blooms. The aerosolization and coastal air concentrations of other common marine phycotoxins have, however, never been examined. This study provides the first (experimental) evidence and characterization of the aerosolization of okadaic acid (OA), homoyessotoxin, and dinophysistoxin-1 using seawater spiked with toxic algae combined with the realistic SSA production in a marine aerosol reference tank (MART). The potential for aerosolization of these phycotoxins was highlighted by their 78- to 1769-fold enrichment in SSAs relative to the subsurface water. To obtain and support these results, we first developed an analytical method for the determination of phycotoxin concentrations in SSAs, which showed good linearity ( R 2 > 0.99), recovery (85.3-101.8%), and precision (RSDs ≤ 17.2%). We also investigated natural phycotoxin air concentrations by means of in situ SSA sampling with concurrent aerosolization experiments using natural seawater in the MART. This approach allowed us to indirectly quantify the (harmless) magnitude of OA concentrations (0.6-51 pg m -3 ) in Belgium's coastal air. Overall, this study provides new insights into the enriched aerosolization of marine compounds and proposes a framework to assess their airborne exposure and effects on human health.

Despite the ubiquitous and persistent presence of microplastic (MP) in marine ecosystems, knowledge of its potential harmful ecological effects is low. In this work, we assessed the risk of floating MP (1 μm-5 mm) to marine ecosystems by comparing ambient concentrations in the global ocean with available ecotoxicity data. The integration of twenty-three species-specific effect threshold concentration data in a species sensitivity distribution yielded a median unacceptable level of 1.21 ∗ 10 5  MP m - ³ (95% CI: 7.99 ∗ 10 3 -1.49 ∗ 10 6  MP m - ³). We found that in 2010 for 0.17% of the surface layer (0-5 m) of the global ocean a threatening risk would occur. By 2050 and 2100, this fraction increases to 0.52% and 1.62%, respectively, according to the worst-case predicted future plastic discharge into the ocean. Our results reveal a spatial and multidecadal variability of MP-related risk at the global ocean surface. For example, we have identified the Mediterranean Sea and the Yellow Sea as hotspots of marine microplastic risks already now and even more pronounced in future decades., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Global change will disturb the frequency, scale and distribution of harmful algal blooms (HABs), but we are unable to predict future HABs due to our limited understanding of how physicochemical changes in the environment affect interspecific competition between dinoflagellates. Trait-based mechanistic modelling is an important tool to unravel and quantify various direct and indirect interactions between species. The present study explores whether MacArthur's consumer-resource model can be used as a viable base model to predict dinoflagellate growth in closed multispecies systems. To this end, two batch culture experiments (294 cultures in total) with monocultures and multispecies cultures of Alexandrium minutum, Prorocentrum lima, P. micans, Protoceratium reticulatum and Scrippsiella trochoidea were performed. Despite changes to the relative (different nitrate concentrations) and absolute nutrient availability (dilutions of L1 medium), P. micans outcompeted all other species in mixed cultures. Consumer-resource modelling parameterized using monoculture growth correctly predicted this species dominance (R² between 0.80 and 0.95). Parameter estimates revealed that P. micans had a faster uptake of nitrogen when compared to its competitors, but did not differ in resource efficiency and natural mortality rate. Yet, while the model accurately predicted community dynamics during the growth phase, it was not able to predict their dynamics beyond the point of quiescence. Consumer-resource modelling was shown to differentiate the roles of resource assimilation, resource efficiency, and natural mortality rates in batch culture experiments with minimal data requirements beyond common measurements. The results suggest that consumer-resource models provide a promising basis for trait-based modelling of interspecific competition between (harmful) algae., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Marine zooplankton are increasingly being affected by recent environmental changes, such as climate change, and respond with profound spatial relocations and shifts in phenology and physiology. In order to predict whether populations are able to persist or adapt to such new conditions, it is essential to understand the molecular basis of such adaptations, which ultimately get translated into these physiological responses. To explore variation in population gene expression across time and space, we investigated transcriptome-level profiles of the calanoid copepod Temora longicornis, that were collected at four different locations in the Belgian Part of the North Sea (BPNS) on three different time points (April, June, October) in 2018. RNA-seq analysis of field collected adults identified large seasonal differences in gene expression, mainly between spring-summer and autumn samples. The largest log-fold changes occurred in a set of genes encoding for ribosomal and myosin (heavy chain) transcripts. Enrichment analysis revealed a strong seasonal pattern in vitellogenin, cuticle and glycolytic gene expression as well. We also found a positive correlation between vitellogenin expression and densities of T. longicornis. No clear spatial variation in expression patterns was found in the BPNS. This study underlines the potential of field gene expression studies for biomonitoring purposes and the significance of considering seasonal variation in future studies., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Water quality standards for cobalt (Co) have not been developed for the European Union or United States. The objective of the present study was to produce freshwater Co toxicity data that could be used by both the European Union and the United States to develop appropriate regulatory standards (i.e., environmental quality standards or predicted-no-effect concentrations in Europe and ambient water quality criteria or state water quality standards in the United States). Eleven species, including algae, an aquatic plant, and several invertebrate and fish species, were used in the performance of acute and chronic Co toxicity tests. Acute median lethal or median effective concentration (LC50 or EC50) values ranged from 90.1 µg Co/L for duckweed (Lemna minor) to 157 000 µg Co/L for midges (Chironomus tentans). Chronic 10% effect concentration (EC10) values ranged from 4.9 µg Co/L for duckweed to 2170 µg Co/L for rainbow trout (Oncorhynchus mykiss). Chronic 20% effect concentration (EC20) values ranged from 11.1 µg Co/L for water flea (Ceriodaphnia dubia) to 2495 µg Co/L for O. mykiss. Results indicated that invertebrate and algae/plant species are more sensitive to chronic Co exposures than fish. Acute-to-chronic ratios (derived as acute LC50s divided by chronic EC20s) were lowest for juvenile O. mykiss (0.6) and highest for the snail Lymnaea stagnalis (2670). Following the European-based approach and using EC10 values, species sensitivity distributions (SSDs) were developed and a median hazardous concentration for 5% of the organisms of 1.80 µg Co/L was derived. Chronic EC20 values were used, also in an SSD approach, to derive a US Environmental Protection Agency-style final chronic value of 7.13 µg Co/L. Environ Toxicol Chem 2020;39:799-811. © 2020 SETAC., (© 2020 SETAC.)

Respiratory exposure to marine phycotoxins is of increasing concern. Inhalation of sea spray aerosols (SSAs), during harmful Karenia brevis and Ostreopsis ovata blooms induces respiratory distress among others. The biogenics hypothesis, however, suggests that regular airborne exposure to natural products is health promoting via a downregulation of the mechanistic target of rapamycin (mTOR) pathway. Until now, little scientific evidence supported this hypothesis. The current explorative in vitro study investigated both health-affecting and potential health-promoting mechanisms of airborne phycotoxin exposure, by analyzing cell viability effects via cytotoxicity assays and effects on the mTOR pathway via western blotting. To that end, A549 and BEAS-2B lung cells were exposed to increasing concentrations (ng·L -1 - mg·L -1 ) of (1) pure phycotoxins and (2) an extract of experimental aerosolized homoyessotoxin (hYTX). The lowest cell viability effect concentrations were found for the examined yessotoxins (YTXs). Contradictory to the other phycotoxins, these YTXs only induced a partial cell viability decrease at the highest test concentrations. Growth inhibition and apoptosis, both linked to mTOR pathway activity, may explain these effects, as both YTXs were shown to downregulate this pathway. This proof-of-principle study supports the biogenics hypothesis, as specific aerosolizable marine products (e.g., YTXs) can downregulate the mTOR pathway., Competing Interests: The authors declare no conflict of interest.

The implementation of cost-effective monitoring programs for zooplankton remains challenging due to the requirements of taxonomical expertise and the high costs of sampling and species identification. To reduce costs, molecular methods have been proposed as alternatives to morphology-based monitoring. Metatranscriptomics can contribute to promote both cost-effectiveness and accuracy of biological assessments of aquatic ecosystems. Here, we describe and evaluate the construction of a metatranscriptome dataset from a pelagic crustacean zooplankton community. We sampled zooplankton in one marine station, named LW02, in the North Sea, in both winter and summer, and generated transcripts using Oxford Nanopore Technology (ONT), a third-generation nanopore-based sequencing technology. ONT is, uniquely, capable of sequencing RNA directly, rather than depending on reverse transcription and PCR, and applicable to be used directly in the field. We found that metatranscriptomics is capable of species detection, including screening for the presence of endoparasites, hence competing with morphological identification. Taxonomic analysis based on ribosomal 18S transcripts identified calanoid copepods, particularly Temora longicornis and Acartia clausi, as the most abundant community members. Moreover, up to 40.4% and 50.5% of all sequences could be assigned to predicted genes in the winter and summer sample, respectively. The most abundant mRNA transcripts with known function coded for essential metabolic processes. GO term annotation revealed that genes involved in glycolytic and translation-related processes were most expressed in the community. Although small in scale, our study provides the basis for future efforts to characterize the metatranscriptome of marine zooplankton communities and its application in biomonitoring programs., Competing Interests: Declaration of competing interest There are no interests to declare., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Sea spray aerosols (SSAs) have profound effects on our climate and ecosystems. They also contain microbiota and biogenic molecules which could affect human health. Yet the exposure and effects of SSAs on human health remain poorly studied. Here, we exposed human lung cancer cells to extracts of a natural sea spray aerosol collected at the seashore in Belgium, a laboratory-generated SSA, the marine algal toxin homoyessotoxin and a chemical inhibitor of the mammalian target of rapamycin (mTOR) pathway. We observed significant increased expression of genes related to the mTOR pathway and Proprotein convertase subtilisin/kexin type 9 (PCSK9) after exposure to homoyessotoxin and the laboratory-generated SSA. In contrast, we observed a significant decrease in gene expression in the mTOR pathway and of PCSK9 after exposure to the natural SSA and the mTOR inhibitor, suggesting induction of apoptosis. Our results indicate that marine biogenics in SSAs interact with PCSK9 and the mTOR pathway and can be used in new potential pharmaceutical applications. Overall, our results provide a substantial molecular evidence base for potential beneficial health effects at environmentally relevant concentrations of natural SSAs.

Understanding the impacts of global change in zooplankton communities is crucial, as alterations in the zooplankton communities can affect entire marine ecosystems. Despite the economic and ecological importance of the calanoid copepod Temora longicornis in the Belgian part of the North Sea, molecular data is still very limited for this species. Using HiSeq Illumina sequencing, we sequenced the whole transcriptome of T. longicornis, after being exposed to realistic temperatures of 14 and 17 °C. After both an acute (1 day) and a more sustained (5 days) thermal exposure to 17 °C, we investigated gene expression differences with animals exposed to 14 °C, which may be critical for the thermal acclimation and resilience of this copepod species. We also studied the possibility of a short term stress recovery of a heat shock. A total of 179,569 transcripts were yielded, of which 44,985 putative ORF transcripts were identified. These transcripts were subsequently annotated into roughly 22,000 genes based on known sequences using Gene Ontology (GO) and KEGG databases. Temora only showed a mild response to both the temperature and the duration of the exposure. We found that the expression of 27 transcripts varied significantly with an increase in temperature of 3 °C, of which eight transcripts were differentially expressed after acute exposure only. Gene set enrichment analysis revealed that, overall, T. longicornis was more impacted by a sustained thermal exposure, rather than an immediate (acute) exposure, with two times as many enriched GO terms in the sustained treatment. We also identified several general stress responses independent of exposure time, such as modified protein synthesis, energy mobilisation, cuticle and chaperone proteins. Finally, we highlighted candidate genes of a possible recovery from heat exposure, identifying similar terms as those enriched in the heat treatments, i.e. related to for example energy metabolism, cuticle genes and extracellular matrix. The data presented in this study provides the first transcriptome available for T. longicornis which can be used for future genomic studies., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

We performed an environmental risk assessment for microplastics (<5 mm) in the marine environment by estimating the order of magnitude of the past, present and future concentrations based on global plastic production data. In 2100, from 9.6 to 48.8 particles m -3 are predicted to float around in the ocean, which is a 50-fold increase compared to the present-day concentrations. From a meta-analysis with effect data available in literature, we derived a safe concentration of 6650 buoyant particles m -3 below which adverse effects are not likely to occur. Our risk assessment (excluding the potential role of microplastics as chemical vectors) suggests that on average, no direct effects of free-floating microplastics in the marine environment are to be expected up to the year 2100. Yet, even today, the safe concentration can be exceeded in sites that are heavily polluted with buoyant microplastics. In the marine benthic compartment between 32 and 144 particles kg -1 dry sediment are predicted to be present in the beach deposition zone. Despite the scarcity of effect data, we expect adverse ecological effects along the coast as of the second half of the 21st century. From then ambient concentrations will start to outrange the safe concentration of sedimented microplastics (i.e. 540 particles kg -1 sediment). Additional ecotoxicological research in which marine species are chronically exposed to realistic environmental microplastic concentration series are urgently needed to verify our findings., (Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.)

It is generally assumed that as long as the majority of species experiences no direct adverse effects attributable to a single substance (i.e., potentially affected fraction [PAF] <5%), no significant structural or functional effects at the community level are expected to occur. Whether this assumption holds for mixed metal contamination is not known. In the present study, we tested this by performing a microcosm experiment in which a naturally occurring freshwater planktonic community was exposed to a copper-nickel-zinc (Cu-Ni-Zn) mixture for 8 wk and various structural and functional community-level traits were assessed. In the low mixture concentration treatments (i.e., Ni-Zn mixtures, because there was no difference in Cu concentrations in these treatments with the control), community-level effects were relatively simple, only involving phytoplankton species groups. In the high mixture concentration treatments (Cu-Ni-Zn mixtures), community-level effects were more complex, involving several phytoplankton and zooplankton species groups. Multisubstance PAF (msPAF) values for all mixture treatments were calculated by applying the concentration addition model to bioavailability-normalized single-metal species sensitivity distributions (SSDs). Consistent effects on the structural traits community composition, abundance of zooplankton species groups, species diversity, and species richness and on the functional trait dissolved organic carbon (DOC) concentration (as a proxy for the microbial loop and pelagic food web interactions) were only observed at msPAF values >0.05 (i.e., in the Cu-Ni-Zn mixture). However, consistent effects on the abundance of various phytoplankton species groups (structural traits) and on 2 measures of community respiration, overnight Δ dissolved oxygen (ΔDO) and ΔpH (functional traits), were already observed at msPAF values of ≤0.05 (i.e., in the Ni-Zn mixture). This indicates that the threshold msPAF value of 0.05 was not protective against metal mixture exposure for all community-level structural and functional endpoints in the present study. A possible explanation for this result is the mismatch between the species in the SSD and those in our microcosm community. Indeed, our data suggest that the presence of one single dominant and very Zn- and/or Ni-sensitive species in the investigated community (i.e., a cyanobacteria of the genus Oscillatoria), which is not represented in the SSD of these metals, was probably the driver of all observed effects at or below an msPAF of 0.05. Overall, the present results show that SSDs are not necessarily a good predictor of community-level effects for all types of communities and that the presence of dominant sensitive species may result in significant, consistent effects on certain structural and functional community-level endpoints at msPAF values ≤0.05, which is generally considered protective in many regulatory frameworks. Environ Toxicol Chem 2018;37:2380-2400. © 2018 SETAC., (© 2018 SETAC.)

Ecotoxicological studies mainly consist of single-species experiments evaluating the effects of a single stressor. However, under natural conditions aquatic communities are exposed to a mixture of stressors. The present study aimed to identify how the toxicity of zinc (Zn) is affected by increased temperature and increased phosphorus (P) supply and how these interactions vary among species, functional groups, and community structure and function. Aquatic microcosms were subjected to 3 Zn concentrations (background, no Zn added, and 75 and 300 μg Zn/L), 2 temperatures (16-19 and 21-24 °C), and 2 different P additions (low, 0.02, and high, 0.4 mg P L -1  wk -1 ) for 5 wk using a full factorial design. During the study, consistent interactions between Zn and temperature were only rarely found at the species level (4%), but were frequently found at the functional group level (36%), for community structure (100%) and for community function (100%; such as dissolved organic carbon concentrations and total chlorophyll). The majority of the Zn × temperature interactions were observed at 300 μg Zn/L and generally indicated a smaller effect of Zn at higher temperature. Furthermore, no clear indication was found that high P addition by itself significantly affected the overall effects of Zn on the community at any level of organization. Interestingly, though, 90% of all the Zn × temperature interactions observed at the species, group, and community composition level were found under high P addition. Collectively, the results of our study with the model chemical Zn suggest that temperature and phosphorus loading to freshwater systems should be accounted for in risk assessment, because these factors may modify the effects of chemicals on the structure and functioning of aquatic communities, especially at higher levels of biological organization. Environ Toxicol Chem 2018;37:2413-2427. © 2018 SETAC., (© 2018 SETAC.)

Understanding how biodiversity (B) affects ecosystem functioning (EF) is essential for assessing the consequences of ongoing biodiversity changes. An increasing number of studies, however, show that environmental conditions affect the shape of BEF relationships. Here, we first use a game-theoretic community model to reveal that a unimodal response of the BEF slope can be expected along environmental stress gradients, but also how the ecological mechanisms underlying this response may vary depending on how stress affects species interactions. Next, we analysed a global dataset of 44 experiments that crossed biodiversity with environmental conditions. Confirming our main model prediction, the effect of biodiversity on ecosystem functioning tends to be greater at intermediate levels of environmental stress, but varies among studies corresponding to differences in stress-effects on species interactions. Together, these results suggest that increases in stress from ongoing global environmental changes may amplify the consequences of biodiversity changes., (© 2018 John Wiley & Sons Ltd/CNRS.)

Human activities increasingly impact the functioning of marine food webs, but anthropogenic stressors are seldom included in ecological study designs. Diet quality, as distinct from just diet quantity, has moreover rarely been highlighted in food web studies in a stress context. We measured the effects of metal and pesticide stress (copper and atrazine) on the contribution of a benthic intertidal diatom community to two processes that are key to the functioning of intertidal systems: biomass (diet quantity) and lipid (diet quality) production. We then examined if stressors affected diatom functioning by selectively targeting the species contributing most to functioning (selective stress effects) or by changing the species' functional contribution (context-dependent effects). Finally, we tested if stress-induced changes in diet quality altered the energy flow to the diatoms' main grazers (harpacticoid copepods). Diatom diet quantity was reduced by metal stress but not by low pesticide levels due to the presence of an atrazine-tolerant, mixotrophic species. Selective effects of the pesticide reduced diatom diet quality by 60% and 75% at low and high pesticide levels respectively, by shifting diatom community structure from dominance by lipid-rich species toward dominance by an atrazine-tolerant, but lipid-poor, species. Context-dependent effects did not affect individual diatom lipid content at low levels of both stressors, but caused diatoms to lose 40% of their lipids at high copper stress. Stress-induced changes in diet quality predicted the energy flow from the diatoms to their copepod consumers, which lost half of their lipids when feeding on diatoms grown under low and high pesticide and high metal stress. Selective pesticide effects were a more important threat for trophic energy transfer than context-dependent effects of both stressors, with shifts in diatom community structure affecting the energy flow to their copepod grazers at stress levels where no changes in diatom lipid content were detected., (© 2018 by the Ecological Society of America.)

Under natural conditions, organisms can experience a variety of abiotic (e.g., temperature, pH) and biotic (e.g., species interactions) factors, which can interact with toxicant effects. By ignoring species interactions conventional ecotoxicological studies (i.e., single-species tests) oversimplify the actual field situation. We investigated whether temperature and interspecific competition affected the effects of zinc (Zn) on a Daphnia longispina population. The D. longispina populations were exposed in a full factorial design to 3 different Zn treatments (background, 29, and 110 μg Zn/L), 2 different temperature regimes (cold, 17-18 °C; warm, 21-22 °C), and 2 interspecific competition levels (no interspecific Brachionus competition = no Brachionus calyciflorus added; interspecific Brachionus competition = B. calyciflorus added). Interspecific Brachionus competition and temperature by itself had a limited effect on the Daphnia abundance but significantly interacted with the highest Zn concentration. Without Brachionus competition the D. longispina juvenile and adult abundances under warm conditions were up to 5.5 and 21 times lower, respectively, in the high Zn treatment in comparison with the Zn control, whereas under cold conditions no significant Zn effect was observed. However, with Brachionus competition the highest Zn treatment was on average 2.2 times less toxic to the D. longispina juvenile abundance at higher temperatures. Under cold conditions the highest Zn treatment affected the juvenile abundance sooner and up to 9 times more negatively when simultaneously faced with Brachionus competition. It is possible that the competition for food reduced the amount of energy that could be used by D. longispina for reproduction, and the metabolic costs increased as a result of Zn stress. The present study clearly illustrated the influence of temperature and competition on the effects of a chemical stressor. Thus, not considering such factors in ecological risk assessment may underestimate or overestimate risks in aquatic ecosystems when extrapolating data from standard single-species tests to the field. Environ Toxicol Chem 2018;37:1668-1678. © 2018 SETAC., (© 2018 SETAC.)

Although metal mixture toxicity has been studied relatively intensely, there is no general consensus yet on how to incorporate metal mixture toxicity into aquatic risk assessment. We combined existing data on chronic metal mixture toxicity at the species level with species sensitivity distribution (SSD)-based in silico metal mixture risk predictions at the community level for mixtures of Ni, Zn, Cu, Cd, and Pb, to develop a tiered risk assessment scheme for metal mixtures in freshwater. Generally, independent action (IA) predicts chronic metal mixture toxicity at the species level most accurately, whereas concentration addition (CA) is the most conservative model. Mixture effects are noninteractive in 69% (IA) and 44% (CA) and antagonistic in 15% (IA) and 51% (CA) of the experiments, whereas synergisms are only observed in 15% (IA) and 5% (CA) of the experiments. At low effect sizes (∼ 10% mixture effect), CA overestimates metal mixture toxicity at the species level by 1.2-fold (i.e., the mixture interaction factor [MIF]; median). Species, metal presence, or number of metals does not significantly affect the MIF. To predict metal mixture risk at the community level, bioavailability-normalization procedures were combined with CA or IA using SSD techniques in 4 different methods, which were compared using environmental monitoring data of a European river basin (the Dommel, The Netherlands). We found that the simplest method, in which CA is directly applied to the SSD (CA SSD ), is also the most conservative method. The CA SSD has median margins of safety (MoS) of 1.1 and 1.2 respectively for binary mixtures compared with the theoretically more consistent methods of applying CA or IA to the dose-response curve of each species individually prior to estimating the fraction of affected species (CA DRC or IA DRC ). The MoS increases linearly with an increasing number of metals, up to 1.4 and 1.7 for quinary mixtures (median) compared with CA DRC and IA DRC , respectively. When our methods were applied to a geochemical baseline database (Forum of European Geological Surveys [FOREGS]), we found that CA SSD yielded a considerable number of mixture risk predictions, even when metals were at background levels (8% of the water samples). In contrast, metal mixture risks predicted with the theoretically more consistent methods (e.g., IA DRC ) were very limited under natural background metal concentrations (<1% of the water samples). Based on the combined evidence of chronic mixture toxicity predictions at the species level and evidence of in silico risk predictions at the community level, a tiered risk assessment scheme for evaluating metal mixture risks is presented, with CA SSD functioning as a first, simple conservative tier. The more complex, but theoretically more consistent and most accurate method, IA DRC , can be used in higher tier assessments. Alternatively, the conservatism of CA SSD can be accounted for deterministically by incorporating the MoS and MIF in the scheme. Finally, specific guidance is also given related to specific issues, such as how to deal with nondetect data and complex mixtures that include so-called data-poor metals. Environ Toxicol Chem 2018;37:623-642. © 2017 SETAC., (© 2017 SETAC.)

Little is known about the effect of metal mixtures on marine organisms, especially after exposure to environmentally realistic concentrations. This information is, however, required to evaluate the need to include mixtures in future environmental risk assessment procedures. We assessed the effect of copper (Cu)-Nickel (Ni) binary mixtures on Mytilus edulis larval development using a full factorial design that included environmentally relevant metal concentrations and ratios. The reproducibility of the results was assessed by repeating this experiment 5 times. The observed mixture effects were compared with the effects predicted with the concentration addition model. Deviations from the concentration addition model were estimated using a Markov chain Monte-Carlo algorithm. This enabled the accurate estimation of the deviations and their uncertainty. The results demonstrated reproducibly that the type of interaction-synergism or antagonism-mainly depended on the Ni concentration. Antagonism was observed at high Ni concentrations, whereas synergism occurred at Ni concentrations as low as 4.9 μg Ni/L. This low (and realistic) Ni concentration was 1% of the median effective concentration (EC50) of Ni or 57% of the Ni predicted-no-effect concentration (PNEC) in the European Union environmental risk assessment. It is concluded that results from mixture studies should not be extrapolated to concentrations or ratios other than those investigated and that significant mixture interactions can occur at environmentally realistic concentrations. This should be accounted for in (marine) environmental risk assessment of metals. Environ Toxicol Chem 2017;36:3471-3479. © 2017 SETAC., (© 2017 SETAC.)

Risk assessment in the European Union implements Zn bioavailability models to derive predicted-no-effect concentrations for Zn. These models are validated within certain boundaries (i.e., pH ≤ 8 and Ca concentrations ≥ 5mg/L), but a substantial fraction of the European surface waters falls outside these boundaries. Therefore, we evaluated whether the chronic Zn biotic ligand model (BLM) for Daphnia magna and the chronic bioavailability model for Pseudokirchneriella subcapitata could be extrapolated to pH > 8 and Ca concentrations < 5 mg/L. Results from D. magna experiments suggested that the BLM is not able to reflect the pH effect over a broad pH range (5.5-8.5). In addition, because of Ca deficiency of D. magna in the soft water tests, we cannot conclude whether the BLM is applicable below its Ca boundary. Results for P. subcapitata experiments showed that the bioavailability model can accurately predict Zn toxicity for Ca concentrations down to 0.8 mg/L and pH values up to 8.5. Because the chronic Zn BLM for D. magna could not be extrapolated beyond its validity boundaries for pH, a generalized bioavailability model (gBAM) was developed. Of 4 gBAMs developed, we recommend the use of gBAM-D, which combines a log-linear relation between the 21-d median effective concentrations (expressed as free Zn 2+ ion activity) and pH, with more conventional BLM-type competition constants for Na, Ca, and Mg. This model is a first step in further improving the accuracy of chronic toxicity predictions of Zn as a function of water chemistry, which can decrease the uncertainty in implementing the bioavailability-based predicted-no-effect concentration in the risk assessment of high-pH and low-Ca concentration regions in Europe. Environ Toxicol Chem 2017;36:2781-2798. © 2017 SETAC., (© 2017 SETAC.)

In the present study, we observed that the chronic Zn Daphnia magna biotic ligand model (BLM) could not accurately predict the chronic Zn toxicity to Ceriodaphnia dubia in a series of (modified) field waters, although the intrinsic sensitivities of the D. magna BLM were specifically calibrated on the sensitivity of C. dubia. Moreover, it was observed that the D. magna BLM underestimated the effect of pH on the toxicity of the free Zn 2+ ion. A preliminary species-specific chronic Zn C. dubia bioavailability model was developed based on the data. The preliminary Zn C. dubia bioavailability model was shown to predict Zn toxicity to C. dubia in a series of natural waters with pH ranging between 7 and 8.3 with reasonable accuracy. The difference in the effect of pH on Zn 2+ toxicity between D. magna and C. dubia (2 phylogenetically closely related species) suggested by our results deserves further attention because of the importance of the cross-species application of bioavailability models in ecological risk-assessment procedures and environmental quality standard derivations for metals. Environ Toxicol Chem 2017;36:2750-2755. © 2017 SETAC., (© 2017 SETAC.)

Although chemical risk assessment is still mainly conducted on a substance-by-substance basis, organisms in the environment are typically exposed to mixtures of substances. Risk assessment procedures should therefore be adapted to fit these situations. Four mixture risk assessment methodologies were compared for risk estimations of mixtures of copper (Cu), zinc (Zn), and nickel (Ni). The results showed that use of the log-normal species sensitivity distribution (SSD) instead of the best-fit distribution and sampling species sensitivities independently for each metal instead of using interspecies correlations in metal sensitivity had little impact on risk estimates. Across 4 different monitoring datasets, between 0% and 52% of the target water samples were estimated to be at risk, but only between 0% and 15% of the target water samples were at risk because of the mixture of metals and not any single metal individually. When a natural baseline database was examined, it was estimated that 10% of the target water samples were at risk because of single metals or their mixtures; when the most conservative method was used (concentration addition [CA] applied directly to the SSD, i.e., CA SSD ). However, the issue of metal mixture risk at geochemical baseline concentrations became relatively small (2% of target water samples) when a theoretically more correct method was used (CA applied to individual dose response curves, i.e., CA DRC ). Finally, across the 4 monitoring datasets, the following order of conservatism for the 4 methods was shown (from most to least conservative, with ranges of median margin of safety [MoS] relative to CA SSD ): CA SSD  > CA DRC (MoS = 1.17-1.25) > IA DRC (independent action (IA) applied to individual dose-response curves; MoS = 1.38-1.60) > IA SSD (MoS = 1.48-1.72). Therefore, it is suggested that these 4 methods can be used in a general tiered scheme for the risk assessment of metal mixtures in a regulatory context. In this scheme, the CA SSD method could serve as a first (conservative) tier to identify situations with likely no potential risk at all, regardless of the method used (the sum toxic unit expressed relative to the 5% hazardous concentration [SumTU HC5 ] < 1) and the IA SSD method to identify situations of potential risk, also regardless of the method used (the multisubstance potentially affected fraction of species using the IA SSD method [msPAF IA,SSD ] > 0.05). The CA DRC and IA DRC methods could be used for site-specific assessment for situations that fall in between (SumTU HC5  > 1 and msPAF IA,SSD  < 0.05). Environ Toxicol Chem 2017;36:2123-2138. © 2017 SETAC., (© 2017 SETAC.)

To improve the ecological relevance of environmental risk assessment, an improved understanding is needed of 1) the influence of environmental conditions on the toxicity of pollutants, and 2) the effect of these factors in combination with possible interpopulation variability. The influences of salinity and dissolved organic carbon (DOC) on the accumulation and effect of copper (Cu) to settled mussels were investigated with mussels from a North Sea and a Baltic Sea population. We found that both populations were equally Cu-sensitive, even though the Baltic Sea population lives in suboptimal conditions. Baltic Sea mussels, however, accumulated more Cu. This suggests that these populations may have different ways of coping with excess Cu. The influence of salinity on Cu toxicity to settled mussels was limited for both populations. An increase in DOC did not decrease the Cu accumulation or effect in either population. This suggests that DOC-Cu complexes are bioavailable for settled mussels. These findings are in contrast with previous research which indicated that DOC decreased the toxicity and accumulation of Cu in the D-larvae life stage. As a consequence, the mussel larval stage is not the most Cu-sensitive life stage at high DOC concentrations. Furthermore, a DOC correction factor for Cu toxicity cannot be used for settled mussels. This should be accounted for in future marine Cu environmental risk assessment. Environ Toxicol Chem 2017;36:2074-2082. © 2017 SETAC., (© 2017 SETAC.)

Estimating ecosystem-level effects from single-species bioassays is a major challenge in environmental risk assessment. Most extrapolation procedures are based on the implicit assumption that species sensitivities are random with regard to their functional traits. Here, we explore how nonrandomness in species sensitivities affects how species-level and ecosystem level effects of chemical exposure correspond. The effect of a correlation between the trait value under control conditions and the sensitivity of the trait to chemical stress is studied for two traits (per capita growth rate and monoculture yield) under constant and temporary exposure. Theoretical model predictions are thereby validated against a 3-week microcosm experiment, in which eight marine diatoms systems with different correlations between trait values and sensitivities were temporary (1 week) or constantly (3 weeks) exposed to two concentrations of the herbicide atrazine (100 and 250 μg L -1 ). Negative correlations increased the reduction in ecosystem functioning (productivity) by atrazine for both traits. However, correlations in the per capita growth rate affected productivity only shortly following changes in environmental conditions, whereas correlations in the monoculture yield affected productivity throughout exposure. Correlations between species sensitivities and functional trait values can thus help to identify when ecosystem-level effects are likely to exceed species-level effects.

Lipophilic marine biotoxins, which are mainly produced by small dinoflagellates, are increasingly detected in coastal waters across the globe. As these producers are consumed by zooplankton and shellfish, the toxins are introduced, bioaccumulated and possibly biomagnified throughout marine food chains. Recent research has demonstrated that ultra-high performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS) is an excellent tool to detect marine toxins in algae and seafood. In this study, UHPLC-HRMS was used to screen lipophilic marine biotoxins in organisms from different trophic levels of the Belgian coastal zone ecosystem. A total of 20 tentatively identified lipophilic compounds was detected. Hereby, the trophic transfer of lipophilic marine biotoxins to the upper trophic level was considered to be rather limited. Furthermore, 36% of the compounds was clearly transferred between different organisms. A significant biotransformation of compounds from the okadaic acid and spirolide toxin groups was observed (64%), mainly in filter feeders. Through a multi-targeted approach, this study showed that marine organisms in the Belgian coastal zone are exposed to a multi-toxin mixture. Further research on both single compound and interactive toxic effects of the frequently detected lipophilic marine toxin ester metabolites throughout the food chain is therefore needed. As a future perspective, confirmatory identification of potential toxins by studying their fragmentation spectra (using new tools such as hybrid quadrupole Q-Exactive™ Orbitrap-MS) is designated., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Although aquatic organisms in the environment are exposed to mixtures of metals, risk assessment for metals is most commonly performed on a metal-by-metal basis. To increase the knowledge about chronic mixture effects, the authors investigated whether metal mixture effects are dependent on the biological species, mixture composition, and metal concentration ratio. The authors evaluated the effects of quaternary Ni-Zn-Cu-Cd and ternary Ni-Zn-Cu mixtures on 48-h algal growth rate (Pseudokirchneriella subcapitata) and 7-d daphnid reproduction (Ceriodaphnia dubia) using a ray design. Single metals were 3-fold to 42-fold more toxic for C. dubia than for P. subcapitata, based on the 50% effective concentration expressed as free metal activity, the range representing different metals. Statistical analysis of mixture effects showed that the ternary and quaternary mixture effects were antagonistic on algal growth relative to the concentration addition (CA) model, when the analysis was based on dissolved concentrations and on free metal ion activities. Using the independent action (IA) model, mixture effects in both rays were statistically noninteractive for algal growth when the analysis was based on dissolved concentrations; however, the interactions shifted toward antagonism when based on free ion activities. The ternary Ni-Zn-Cu mixture acted antagonistically on daphnid reproduction relative to both reference models, either expressed as free ion activities or dissolved concentrations. When Cd was added to the mixture, however, the mixture effects shifted toward noninteractivity for daphnids. The metal concentration ratio did not significantly influence the magnitude of observed antagonistic effects. Regardless of statistical interactions observed, based on the present study, CA and in most instances also IA can serve as a protective model for ternary Ni-Zn-Cu and quaternary Ni-Zn-Cu-Cd toxicity to both species. Environ Toxicol Chem 2017;36:1056-1066. © 2016 SETAC., (© 2016 SETAC.)

Little is known about the influence that environmental stressors may have on genome-wide methylation patterns, and to what extent epigenetics may be involved in environmental stress response. Yet, studies of methylation patterns under stress could provide crucial insights on stress response and toxicity pathways. Here, we focus on genome-wide methylation patterns in the microcrustacean Daphnia magna, a model organism in ecotoxicology and risk assessment, exposed to the toxic cyanobacterium Microcystis aeruginosa. Bisulfite sequencing of exposed and control animals highlighted differential methylation patterns in Daphnia upon exposure to Microcystis primarily in exonic regions. These patterns are enriched for serine/threonine amino acid codons and genes related to protein synthesis, transport and degradation. Furthermore, we observed that genes with differential methylation corresponded well with genes susceptible to alternative splicing in response to Microcystis stress. Overall, our results suggest a complex mechanistic response in Daphnia characterized by interactions between DNA methylation and gene regulation mechanisms. These results underscore that DNA methylation is modulated by environmental stress and can also be an integral part of the toxicity response in our study species.

Recently, several bioavailability-based models have been shown to predict acute metal mixture toxicity with reasonable accuracy. However, the application of such models to chronic mixture toxicity is less well established. Therefore, we developed in the present study a chronic metal mixture bioavailability model (MMBM) by combining the existing chronic daphnid bioavailability models for Ni, Zn, and Pb with the independent action (IA) model, assuming strict non-interaction between the metals for binding at the metal-specific biotic ligand sites. To evaluate the predictive capacity of the MMBM, chronic (7d) reproductive toxicity of Ni-Zn-Pb mixtures to Ceriodaphnia dubia was investigated in four different natural waters (pH range: 7-8; Ca range: 1-2 mM; Dissolved Organic Carbon range: 5-12 mg/L). In each water, mixture toxicity was investigated at equitoxic metal concentration ratios as well as at environmental (i.e. realistic) metal concentration ratios. Statistical analysis of mixture effects revealed that observed interactive effects depended on the metal concentration ratio investigated when evaluated relative to the concentration addition (CA) model, but not when evaluated relative to the IA model. This indicates that interactive effects observed in an equitoxic experimental design cannot always be simply extrapolated to environmentally realistic exposure situations. Generally, the IA model predicted Ni-Zn-Pb mixture toxicity more accurately than the CA model. Overall, the MMBM predicted Ni-Zn-Pb mixture toxicity (expressed as % reproductive inhibition relative to a control) in 85% of the treatments with less than 20% error. Moreover, the MMBM predicted chronic toxicity of the ternary Ni-Zn-Pb mixture at least equally accurately as the toxicity of the individual metal treatments (RMSE Mix  = 16; RMSE Zn only  = 18; RMSE Ni only  = 17; RMSE Pb only  = 23). Based on the present study, we believe MMBMs can be a promising tool to account for the effects of water chemistry on metal mixture toxicity during chronic exposure and could be used in metal risk assessment frameworks., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The univariate effects of Ca, pH, and dissolved organic carbon (DOC) on chronic (48-h) Pb toxicity to the freshwater rotifer Brachionus calyciflorus were investigated. High pH (8.2) and higher concentrations of DOC were protective against filtered Pb toxicity, whereas Ca was not. However, expressed as the free Pb 2+ ion, Pb 2+ toxicity increased with increasing DOC concentration, indicating that Pb-fulvic acid (FA) complexes may be bioavailable and may contribute to toxicity. Two different bioavailability models were developed. The pH effect was modeled as a single-site competition effect by H + (log K HBL  = 7.14). In the first model, only this H + effect was considered; in a second model, a log-linear fulvic acid bioavailability effect (S FA  = 0.602) was also incorporated. Both models predicted chronic Pb toxicity for most waters used for model development within a 2-fold error. To validate the predictive capacities of the models, chronic Pb toxicity to B. calyciflorus was tested in 5 natural waters and a reference water. Both models consistently underestimated Pb toxicity in the natural waters, as a result of a shift in rotifer sensitivity to Pb between the development and the validation test series. However, optimizing the intrinsic sensitivity in the models specific for the validation test series resulted in reasonable predictions of Pb toxicity in the natural waters. The second, more complex model predicted chronic Pb toxicity most accurately. The protective effects of water chemistry on Pb toxicity to Brachionus are comparable to those observed for Ceriodaphnia. However, the developed Brachionus models were not able to accurately predict chronic Pb toxicity to the rotifer Philodina rapida. It remains unclear why the influence of water chemistry on chronic Pb toxicity appears to be different between 2 rotifer species. Environ Toxicol Chem 2016;35:2977-2986. © 2016 SETAC., (© 2016 SETAC.)

There is now ample evidence that biodiversity stabilizes aggregated ecosystem functions, such as primary production, in changing environments. In primary producer systems, this stabilizing effect is found to be driven by higher functional resistance (i.e., reduced changes in functions by environmental changes) rather than through higher functional resilience (i.e., rapid recovery following environmental changes) in more diverse systems. The stability of aggregated ecosystem functions directly depends on changes in species composition and by consequence their functional contributions to ecosystem functions. Still, it remains only theoretically explored how biodiversity can stabilize ecosystem functions by affecting compositional stability. Here, we demonstrate how biodiversity effects on compositional stability drive biodiversity effects on functional stability in diatom communities. In a microcosm experiment, we exposed 39 communities of five different levels of species richness (1, 2, 4, 6, and 8 species) to three concentrations of a chemical stressor (0, 25, and 250 μg/L atrazine) for four weeks, after which all communities were transferred to atrazine-free medium for three more weeks. Biodiversity simultaneously increased, increasing functional and compositional resistance, but decreased functional and compositional resilience. These results confirm the theoretically proposed link between biodiversity effects on functional and compositional stability in primary producer systems, and provide a mechanistic underpinning for observed biodiversity-stability relationships. Finally, we discuss how higher compositional stability can be expected to become increasingly important in stabilizing ecosystem functions under field conditions when multiple environmental stressors fluctuate simultaneously., (© 2016 by the Ecological Society of America.)

A major problem with risk assessment of chemicals is the extrapolation of laboratory single-species toxicity tests, which oversimplify the actual field situation by ignoring species interactions, to natural communities. The authors tested if the bioavailability-normalized 5% hazardous concentration (HC5) estimated from chronic planktonic single-species toxicity data (HC5 plankton ) for zinc (Zn) is protective for a plankton community and investigated the direct and indirect effects of Zn (at HC5 and HC50) on a freshwater community's structure and function. Microcosms were exposed to 3 different Zn concentrations (background, HC5 plankton  = 75 μg Zn/L and HC50 plankton  = 300 μg Zn/L) for 5 wk. The planktonic groups revealed a consistent no-observed-effect concentration for the community of 75 μg Zn/L, similar to or higher than the HC5 plankton , thus suggesting its protectiveness in the present study. At 300 μg Zn/L a significant reduction in cladocerans resulted in increases of rotifer, ciliate, and phytoplankton abundance. In addition, the phytoplankton community shifted in dominance from grazing-resistant to edible species. Contrary to the species sensitivity distribution (SSD) prediction, which identified phytoplankton as the most sensitive group, only the total chlorophyll and the abundance of 2 phytoplankton species were adversely affected at 300 μg Zn/L. Thus, although the HC5 estimated from the bioavailability-normalized SSD was overall protective for the plankton community, the SSD was not able to correctly predict the species sensitivity ranking within their community context at the HC50. Environ Toxicol Chem 2016;35:2698-2712. © 2016 SETAC., (© 2016 SETAC.)

Harmful algal blooms (HABs) and marine pathogens - like Vibrio spp. - are increasingly common due to climate change. These stressors affect the growth, viability and development of bivalve larvae. Little is known, however, about the potential for interactions between these two concurrent stressors. While some mixed exposures have been performed with adult bivalves, no such work has been done with larvae which are generally more sensitive. This study examines whether dinoflagellates and bacteria may interactively affect the viability and immunological resilience of blue mussel Mytilus edulis larvae. Embryos were exposed to environmentally relevant concentrations (100, 500, 2500 & 12,500 cells ml(-1)) of a dinoflagellate (Alexandrium minutum, Alexandrium ostenfeldii, Karenia mikimotoi, Protoceratium reticulatum, Prorocentrum cordatum, P. lima or P. micans), a known pathogen (Vibrio coralliilyticus/neptunius-like isolate or Vibrio splendidus; 10(5) CFU ml(-1)), or both. After five days of exposure, significant (p < 0.05) adverse effects on larval viability and larval development were found for all dinoflagellates (except P. cordatum) and V. splendidus. Yet, despite the individual effect of each stressor, no significant interactions were found between the pathogens and harmful algae. The larval viability and the phenoloxidase innate immune system responded independently to each stressor. This independence may be related to a differential timing of the effects of HABs and pathogens., (Copyright © 2016 Elsevier Ltd. All rights reserved.)