Resilience of a microphytobenthos community from the Severn Estuary, UK, to chlorination: A mesocosm approach.

The Severn Estuary is a large macrotidal estuary which includes an extensive mudflat with microphytobenthos (MPB) playing a key role in the ecosystem. This study evaluated the impact of chlorination at two different dosing levels (0.05 and 0.5 mg/l as total residual oxidants, TRO, representative of potential concentrations in the mixing zone and within the cooling water systems of a power station) on a MPB community representative of the Severn Estuary. Biomass and diversity were not negatively impacted while physiology was partially affected at the beginning of the experiment, and it recovered towards the end of the experiment. Further investigations for diversity are needed to consolidate our findings. In conclusion our results show that MPB is resilient to chlorination up to a concentration of 0.5 mg/l which is much higher (>10 times) than what might be expected near the chlorinated discharges for most coastal power stations.

The behavioural response of European seabass (Dicentrarchus labrax) to chlorinated seawater effluents.

Seawater chlorination is widely used for coastal, marine industries for the prevention of fouling. Using a choice chamber system, we investigated the influence of chlorinated seawater at typical concentrations occurring near chlorinated cooling water discharges, on the behaviour of juvenile seabass (Dicentrachus labrax). These studies showed that there was evidence of an effect of chlorination, with models predicting 22% fewer fish present in the chlorine dosed chamber compared to the undosed chamber in all control runs (mean number of fish in the dosed chamber in all control runs was 2.23 in comparison to 1.74 when chlorine was present). These data suggest that when food is available D. labrax will enter areas that are at mean chlorination levels of 0.04 mg l-1, which are typical close to the discharge of power station cooling water but are more likely to move away once the food supply is reduced.

Chlorination in power station cooling water systems: Effect on biomass, abundance and physiology of natural phytoplankton communities.

Chlorination is a widely used method to prevent biofouling in power station cooling water systems in coastal and estuarine environments. This study evaluated the impact of chlorination together with temperature increase to simulate primary entrainment of a phytoplankton community. Biomass, diversity, and photosynthetic activity were monitored over 72 hours to establish impacts on the phytoplankton community. Biomass was significantly reduced after treatment. The mean cell size of the population significantly increased immediately after treatment highlighting an impact on the smaller cell size species of the community (picophytoplankton). Changes in accessory pigments composition suggest an effect on groups such as Prasinophyceae, Cyanobacteria and Chlorophycea. Species composition, dominated by diatoms, was also affected with Skeletonema marinoi and Asterionellopsis glacialis amongst the most sensitive species. Photosynthetic activity was affected in the short term but recovered after 48 hours. This study shows that by using a combination of measurements (e.g biomass, diversity, and physiology) the effects of entrainment in power station cooling water systems, that may be of longer-term significance for specific functional groups of phytoplankton communities, can be discerned. These changes would not necessarily be seen using individual techniques alone such as cell number counts or biomass assessment which may indicate apparent community recovery.

Key Opportunities to Replace, Reduce, and Refine Regulatory Fish Acute Toxicity Tests.

Fish acute toxicity tests are conducted as part of regulatory hazard identification and risk-assessment packages for industrial chemicals and plant protection products. The aim of these tests is to determine the concentration which would be lethal to 50% of the animals treated. These tests are therefore associated with suffering in the test animals, and Organisation for Economic Co-operation and Development test guideline 203 (fish, acute toxicity) studies are the most widely conducted regulatory vertebrate ecotoxicology tests for prospective chemical safety assessment. There is great scope to apply the 3Rs principles-the reduction, refinement, and replacement of animals-in this area of testing. An expert ecotoxicology working group, led by the UK National Centre for the Replacement, Refinement and Reduction of Animals in Research, including members from government, academia, and industry, reviewed global fish acute test data requirements for the major chemical sectors. The present study highlights ongoing initiatives and provides an overview of the key challenges and opportunities associated with replacing, reducing, and/or refining fish acute toxicity studies-without compromising environmental protection. Environ Toxicol Chem 2020;39:2076-2089. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Chlorine toxicity to Navicula pelliculosa and Achnanthes spp. in a flow-through system: The use of immobilised microalgae and variable chlorophyll fluorescence.

Chlorination is a widely used antifouling method for freshwater and marine applications. Chlorine added to seawater reacts to form oxidants that are toxic to biofouling organisms. Further, the oxidants that result are short-lived, but may nevertheless affect non-target species in waterbodies receiving the antifouling effluent. This study evaluated the toxicity of chlorinated seawater (e.g. following sodium hypochlorite addition) on two different species of marine benthic diatoms (Achnanthes spp., and Navicula pelliculosa), which are representative of microphytobenthos communities - an important component in coastal habitats that may be exposed to chlorinated seawater. To evaluate the growth inhibition over a 72 h period, algae were immobilised in alginate beads and exposed to different levels of chlorination in a flow through system. Growth rates and physiological condition of the microalgae were evaluated using a Fast Repetition Rate fluorometer (FRRf). To determine whether alginate influenced the sensitivity of algal response, studies were also conducted in a static test system (without renewal of test solutions) using both free cells and immobilised cells with initial chlorine added to achieve a similar range of concentrations as those used in the flow-through study. Within the first hour of the exposure period there was an indication that, for both species, the free algal cells in the static system were more sensitive to exposure to chlorinated seawater than were alginate-immobilised cells in the flow through system. Immobilised cells in a static system with a single addition of chlorine were also less sensitive to chlorination than free algal cells. However, for periods of 24 h or more due to decay of TRO in the static system the exposure of immobilised algae in the flow through system had a greater impact and hence lower effect concentrations. For the flow-through studies Achnanthes spp. was the most sensitive after 72 h exposure with a potential no effect concentration EC10 value of 0.02 mg l-1 as Cl2 equivalents expressed as total residual oxidants (TRO) compared 0.04 mg l-1 TRO for N. pelliculosa. Immobilisation of algal cells in alginate was found to be an effective means of determining the impact of chlorination and is likely to be effective for other non-persistent substances. Based on the data produced, the extent and significance of ecological effects of chlorination upon algal species typical of microphytobenthos are likely to be limited providing discharges comply with a maximum allowable concentration of 0.01 mg l-1 TRO at the edge of an agreed mixing zone.

Advancing the 3Rs in regulatory ecotoxicology: A pragmatic cross-sector approach.

The ecotoxicity testing of chemicals for prospective environmental safety assessment is an area in which a high number of vertebrates are used across a variety of industry sectors. Refining, reducing, and replacing the use of animals such as fish, birds, and amphibians for this purpose addresses the ethical concerns and the increasing legislative requirements to consider alternative test methods. Members of the UK-based National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs) Ecotoxicology Working Group, consisting of representatives from academia, government organizations, and industry, have worked together over the past 6 y to provide evidence bases to support and advance the application of the 3Rs in regulatory ecotoxicity testing. The group recently held a workshop to identify the areas of testing, demands, and drivers that will have an impact on the future of animal use in regulatory ecotoxicology. As a result of these discussions, we have developed a pragmatic approach to prioritize and realistically address key opportunity areas, to enable progress toward the vision of a reduced reliance on the use of animals in this area of testing. This paper summarizes the findings of this exercise and proposes a pragmatic strategy toward our key long-term goals-the incorporation of reliable alternatives to whole-organism testing into regulations and guidance, and a culture shift toward reduced reliance on vertebrate toxicity testing in routine environmental safety assessment. Integr Environ Assess Manag 2016;12:417-421. © 2015 SETAC.

Ecological modelling and toxicity data coupled to assess population recovery of marine amphipod Gammarus locusta: Application to disturbance by chronic exposure to aniline.

A population agent-based model of marine amphipod Gammarus locusta was designed and implemented as a basis for ecological risk assessment of chemical pollutants impairing life-history traits at the individual level. We further used the model to assess the toxic effects of aniline (a priority hazardous and noxious substance, HNS) on amphipod populations using empirically-built dose-response functions derived from a chronic bioassay that we previously performed with this species. We observed a significant toxicant-induced mortality and adverse effects in reproductive performance (reduction of newborn production) in G. locusta at the individual level. Coupling the population model with the toxicological data from the chronic bioassay allowed the projection of the ecological costs associated with exposure to aniline that might occur in wild populations. Model simulations with different scenarios indicated that even low level prolonged exposure to the HNS aniline can have significant long-term impacts on G. locusta population abundance, until the impacted population returns to undisturbed levels. This approach may be a useful complement in ecotoxicological studies of chemical pollution to transfer individual-collected data to ecological-relevant levels.

Principles of sound ecotoxicology.

We have become progressively more concerned about the quality of some published ecotoxicology research. Others have also expressed concern. It is not uncommon for basic, but extremely important, factors to apparently be ignored. For example, exposure concentrations in laboratory experiments are sometimes not measured, and hence there is no evidence that the test organisms were actually exposed to the test substance, let alone at the stated concentrations. To try to improve the quality of ecotoxicology research, we suggest 12 basic principles that should be considered, not at the point of publication of the results, but during the experimental design. These principles range from carefully considering essential aspects of experimental design through to accurately defining the exposure, as well as unbiased analysis and reporting of the results. Although not all principles will apply to all studies, we offer these principles in the hope that they will improve the quality of the science that is available to regulators. Science is an evidence-based discipline and it is important that we and the regulators can trust the evidence presented to us. Significant resources often have to be devoted to refuting the results of poor research when those resources could be utilized more effectively.

Ocean acidification increases the toxicity of contaminated sediments.

Ocean acidification (OA) may alter the behaviour of sediment-bound metals, modifying their bioavailability and thus toxicity. We provide the first experimental test of this hypothesis with the amphipod Corophium volutator. Amphipods were exposed to two test sediments, one with relatively high metals concentrations (Σmetals 239 mg kg(-1) ) and a reference sediment with lower contamination (Σmetals 82 mg kg(-1) ) under conditions that mimic current and projected conditions of OA (390-1140 µatm pCO2 ). Survival and DNA damage was measured in the amphipods, whereas the flux of labile metals was measured in the sediment and water column (WC) using Diffusive Gradients in Thin-films. The contaminated sediments became more acutely toxic to C. volutator under elevated pCO2 (1140 µatm). There was also a 2.7-fold increase in DNA damage in amphipods exposed to the contaminated sediment at 750 µatm pCO2 , as well as increased DNA damage in organisms exposed to the reference sediment, but only at 1140 µatm pCO2 . The projected pCO2 concentrations increased the flux of nickel and zinc to labile states in the WC and pore water. However, the increase in metal flux at elevated pCO2 was equal between the reference and contaminated sediments or, occasionally, greater from reference sediments. Hence, the toxicological interaction between OA and contaminants could not be explained by e ffects of pH on metal speciation. We propose that the additive physiological effects of OA and contaminants will be more important than changes in metal speciation in determining the responses of benthos to contaminated sediments under OA. Our data demonstrate clear potential for near-future OA to increase the susceptibility of benthic ecosystems to contaminants. Environmental policy should consider contaminants within the context of changing environmental conditions. Specifically, sediment metals guidelines may need to be reevaluated to afford appropriate environmental protection under future conditions of OA.

Exposure of sticklebacks (Gasterosteus aculeatus) to cadmium sulfide nanoparticles: biological effects and the importance of experimental design.

Effects of nanoparticles on aquatic organisms have been little studied to date and toxicological data are urgently needed for development of regulatory frameworks for these substances. Here, we report the findings of a study exposing sticklebacks to cadmium sulfide (CdS) as bulk material and quantum dots. Fish were exposed for 21 d in a flow through test system to 5, 50 or 500 microg l(-1) CdS nanoparticles (nCdS) coated in thiol terminated methyl polyethylene glycol (MPEG), bulk CdS or MPEG at 500 microg l(-1) (nominal concentrations). With the exception of the highest nCdS exposure, measured concentrations were approximately one order of magnitude below nominal. A single fish from each group (excluding MPEG) was examined using energy dispersive X-ray analysis (EDX) to localise cadmium, however, cadmium could not be detected in whole body sections. Elevated levels of oxidized glutathione were measured in the gills of fish exposed to 50 and 500 microg l(-1) nCdS. Induction of vitellogenin synthesis was not detected in any of the treatment groups. The number of males engaged in nest-building behaviour following exposure to 500 microg l(-1) nCdS was reduced and livers of 4/6 fish in the same treatment displayed hepatocellular nuclear pleomorphism. The results are discussed emphasising the fundamental importance of experimental design and the need to understand the behaviour of nanoparticles in the aqueous phase.