Using metabolomics to assess the sub-lethal effects of zinc and boscalid on an estuarine polychaete worm over time.

INTRODUCTION: Zinc is a heavy metal commonly detected in urban estuaries around Australia. Boscalid is a fungicide found in estuaries, both in water and sediment, it enters the system predominantly through agricultural run-off. Zinc is persistent while boscalid breaks down, with a half-life of 108 days. Both contaminants are widely distributed and their effects on ecosystems are not well understood. OBJECTIVES: The aim of this study was to determine the metabolite changes in Simplisetia aequisetis (an estuarine polychaete) following laboratory exposure to a sub-lethal concentration of zinc or boscalid over a 2-week period. METHODS: Individuals were collected at six time points over a 2-week period. Whole polychaete metabolites were extracted and quantified using a multi-platform approach. Polar metabolites were detected using a semi-targeted GC-MS analysis and amine containing compounds were analysed using a targeted LC-MS analysis. Total lipid energy content was also analysed for Simplisetia aequisetis. RESULTS: The pathways that responded to zinc and boscalid exposure were alanine, aspartate and glutamate metabolism (AAG); glycine, serine and threonine metabolism (GST) and metabolites associated with the tricarboxylic acid cycle (TCA). Results showed that changes in total abundance of some metabolites could be detected as early as 24-h exposure. Changes were detected in the metabolites before commonly used total lipid energy assays identified effects. CONCLUSION: A multi-platform approach provided a holistic overview of the metabolomic response to contaminants in polychaetes. This approach shows promise to be used in biomonitoring programs to provide early diagnostic indicators of contamination and exposure.

Metabolite Changes in an Estuarine Annelid Following Sublethal Exposure to a Mixture of Zinc and Boscalid.

Environmental pollutants such as heavy metals and fungicides pose a serious threat to waterways worldwide. Toxicological assessment of such contaminants is usually conducted using single compound exposures, as it is challenging to understand the effect of mixtures on biota using standard ecotoxicological methods; whereas complex chemical mixtures are more probable in ecosystems. This study exposed Simplisetia aequisetis (an estuarine annelid) to sublethal concentrations of a metal (zinc) and a fungicide (boscalid), both singly and as a mixture, for two weeks. Metabolomic analysis via gas and liquid chromatography-mass spectrometry was used to measure the stress response(s) of the organism following exposure. A total of 75 metabolites, including compounds contributing to the tricarboxylic acid cycle, the urea cycle, and a number of other pathways, were identified and quantified. The multiplatform approach identified distinct metabolomic responses to each compound that differed depending on whether the substance was presented singly or as a mixture, indicating a possible antagonistic effect. The study demonstrates that metabolomics is able to elucidate the effects and mode of action of contaminants and can identify possible outcomes faster than standard ecotoxicological endpoints, such as growth and reproduction. Metabolomics therefore has a possible future role in biomonitoring and ecosystem health assessments.

Towards a general framework for the assessment of interactive effects of multiple stressors on aquatic ecosystems: Results from the Making Aquatic Ecosystems Great Again (MAEGA) workshop.

A workshop was held in Wageningen, The Netherlands, in September 2017 to collate data and literature on three aquatic ecosystem types (agricultural drainage ditches, urban floodplains, and urban estuaries), and develop a general framework for the assessment of multiple stressors on the structure and functioning of these systems. An assessment framework considering multiple stressors is crucial for our understanding of ecosystem responses within a multiply stressed environment, and to inform appropriate environmental management strategies. The framework consists of two components: (i) problem identification and (ii) impact assessment. Both assessments together proceed through the following steps: 1) ecosystem selection; 2) identification of stressors and quantification of their intensity; 3) identification of receptors or sensitive groups for each stressor; 4) identification of stressor-response relationships and their potential interactions; 5) construction of an ecological model that includes relevant functional groups and endpoints; 6) prediction of impacts of multiple stressors, 7) confirmation of these predictions with experimental and monitoring data, and 8) potential adjustment of the ecological model. Steps 7 and 8 allow the assessment to be adaptive and can be repeated until a satisfactory match between model predictions and experimental and monitoring data has been obtained. This paper is the preface of the MAEGA (Making Aquatic Ecosystems Great Again) special section that includes three associated papers which are also published in this volume, which present applications of the framework for each of the three aquatic systems.

What are the roadblocks to using population models in ecotoxicology studies?

Understanding how pollution affects populations is critical for targeted environmental risk assessments and adequate protection of the environment. However, the vast majority of ecotoxicology studies still have a traditional focus of identifying effects on individual organisms and do not measure the effects at the population-level. Modelling tools that measure population effects of pollution are available and would add value to current ecotoxicology studies by aligning outcomes more closely to what needs to be protected. In this paper I outline three possible reasons why this knowledge gap still exists and consider how they could be adopted more broadly, including better considerations about what endpoints should be measured at the initial study design phase. The purpose of highlighting this knowledge gap is to assist in facilitating the integration of population-level endpoints into routine pollution monitoring programs and progress of ecologically relevant ecotoxicology research.

Ecological responses to contamination: a meta-analysis of experimental marine studies.

Identifying general response patterns to contamination in the environment is critical for reliable assessments of ecosystem health. However, it is not often clear if there are biases in the information used to understand general effects of contamination. To investigate this we conducted a review of 314 studies that experimentally manipulated contaminants and measured the effects on marine invertebrate taxa. The majority of studies investigated the effects of metals (54%) on individual taxa (mainly bivalves, amphipods, copepods). Ecologically relevant responses to contamination were measured in only 22% of the studies. A meta-analysis using studies that measured ecological responses to copper illustrated a general negative effect of copper and highlighted the bias towards field or laboratory experiments that measure community or individual-level responses. There is a need for diversification of studies that investigate the ecological effects of contamination as an important advancement in ecotoxicology and ecological research and environmentally relevant risk assessments.

Detecting benthic community responses to pollution in estuaries: a field mesocosm approach.

Biological stress responses in individuals are used as indicators of pollution in aquatic ecosystems, but detecting ecologically relevant responses in whole communities remains a challenge. We developed an experimental approach to detect the effects of pollution on estuarine communities using field-based mesocosms. Mesocosms containing defaunated sediments from four estuaries in southeastern Australia that varied in sediment contamination were transplanted and buried in sediments of the same four estuaries for six weeks. Mesocosm sediment properties and metal concentrations remained representative of their source locations. In each estuary, fauna communities associated with sediments derived from the site with the highest metal concentrations were significantly different from other communities. This pattern was evident for some of the individual taxa, in particular the polychaete Capitella sp. Consistent responses across estuaries suggest numbers of individuals, and especially Capitella sp., could be used to identify contaminated sediments in estuaries with similar fauna and site characteristics.