Groundwater is a hidden global keystone ecosystem.

Groundwater is a vital ecosystem of the global water cycle, hosting unique biodiversity and providing essential services to societies. Despite being the largest unfrozen freshwater resource, in a period of depletion by extraction and pollution, groundwater environments have been repeatedly overlooked in global biodiversity conservation agendas. Disregarding the importance of groundwater as an ecosystem ignores its critical role in preserving surface biomes. To foster timely global conservation of groundwater, we propose elevating the concept of keystone species into the realm of ecosystems, claiming groundwater as a keystone ecosystem that influences the integrity of many dependent ecosystems. Our global analysis shows that over half of land surface areas (52.6%) has a medium-to-high interaction with groundwater, reaching up to 74.9% when deserts and high mountains are excluded. We postulate that the intrinsic transboundary features of groundwater are critical for shifting perspectives towards more holistic approaches in aquatic ecology and beyond. Furthermore, we propose eight key themes to develop a science-policy integrated groundwater conservation agenda. Given ecosystems above and below the ground intersect at many levels, considering groundwater as an essential component of planetary health is pivotal to reduce biodiversity loss and buffer against climate change.

Monitoring Groundwater Health Using Citizen Scientists in Semi-Arid Regional Australia.

Citizen science (CS) around the world is undergoing a resurgence, potentially due to the utilization of new technologies and methods to capture information, such as data and photo entry via mobile phone apps. CS has been used in aquatic ecology for several decades, however the use of volunteers to collect data in groundwaters has rarely occurred. Groundwater research, particularly groundwater ecosystems, is unevenly distributed across the world, limiting our knowledge of these ecosystems and their functions. Here, we engaged six volunteer farmers in semi-arid region of north-western New South Wales, Australia to participate in an assessment of groundwater health using privately owned wells. Volunteers were supplied with sampling kits and instructions on sampling methods. Data retrieved indicated the health of the groundwater ecosystems, simultaneously providing information on water quality and groundwater biota present within the farm aquifers. Diverse stygofauna were collected from the trial, which reflected historical records of stygofauna within the same catchment indicating the viability of using citizen scientist for data collection. The citizen science project not only aided the collection of data and assessment of groundwater health, but also provided a tool for education, attracting media attention which furthered the education to a national audience. The amount of data still required to understand groundwater ecosystems, combined with the urgency to manage these environments, suggests that citizen scientists may complement the efforts of scientists around the globe to establish the impacts and consequences of human activities on this resource.

STYGOTOX: A Quality-Assessed Database of (Eco)Toxicological Data on Stygofauna and Other Aquatic Subterranean Organisms.

We have compiled the toxicity data on stygofauna and other aquatic subterranean organisms in one (eco)toxicological database. A total of 46 studies were found, containing 472 toxic endpoints covering 43 different stressors. These compounds were tested on subterranean organisms from four phyla, 12 orders, 24 genera, and 55 species. The studies included were published between 1976 and December 2023 using fauna collected in 13 different countries. The suitability of the studies was assessed to indicate the completeness of reporting and their suitability for use in hazard and risk assessment. This compilation provides a valuable source of data for future development of toxicity testing protocols for groundwater organisms, and to support decision-making, ecological risk assessments and the derivation of water quality criteria for the protection of groundwater ecosystems. Environ Toxicol Chem 2024;00:1-9. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Urbanisation of floodplain ecosystems: Weight-of-evidence and network meta-analysis elucidate multiple stressor pathways.

Freshwater floodplains are dynamic, diverse ecosystems that represent important transition zones between terrestrial, riparian, subsurface and aquatic habitats. Given their historic importance in human development, floodplains have been exposed to a variety of pressures, which in combination have been instrumental in driving changes within these ecosystems. Here, we present an evidence-based framework to explore direct and indirect effects of pressures and stressors on floodplain ecosystems and test this structure within the urban landscape. Evidence was obtained from peer-reviewed scientific literature, focusing on effects of key pressures and stressors on receptors, including species composition (e.g., species presence-absence, diversity) and ecosystem function (e.g., biomass, decomposition). The strength of direct and indirect effects of individual and multiple stressors on biological receptors was quantified using two separate analyses: an evidence-weighted analysis and a quantitative network meta-analysis using data extracted from 131 studies. Results demonstrate the power of adopting a systematic framework to advance quantitative assessment of floodplain ecosystems affected by multiple stressors. While direct pathways were generally stronger and provided the core network skeleton, there were many more significant indirect pathways indicating evidence gaps in our mechanistic understanding of these processes. Indeed, the importance of indirect pathways (e.g. increase in impervious surface → increase in the accumulation rate of sediment nutrients) suggests that embracing complexity in network meta-analysis is a necessary step in revealing a more complete snapshot of the network. Results from the weight-of-evidence approach generally mirrored the direct pathway structure and demonstrated the strength of incorporating study quality alongside data sufficiency. Networks illustrated novel disturbance pathways (e.g., decrease in habitat structure → decrease in structure and function of aquatic and riparian assemblages) that can be used for hypothesis generation for future scientific enquiries. Our results highlight the broader applicability of adopting the proposed framework for assessing complex environments, such as floodplains.

Recommendations for ecotoxicity testing with stygobiotic species in the framework of groundwater environmental risk assessment.

As a consequence of the growing global dependence on groundwater resources, environmental risk assessments (ERA) for groundwater are increasingly required and, with that, ecotoxicological studies with groundwater fauna (stygofauna). However, the literature on the ecotoxicological studies with stygobiotic species (i.e. species that complete their life cycle exclusively in groundwater) has not expanded significantly since the first paper published in this field. The limitations regarding the use of stygobiotic species for ecotoxicological testing are clear and consistent across the globe; stygobiotic species are often 1) naturally present in low numbers, 2) difficult to collect, and 3) difficult to culture under laboratory conditions. This paper reviews the methods used in ecotoxicological studies performed with stygobiotic species, and provides ten recommendations for Good Laboratory Practice (GLP) for such tests. The recommendations focused on the following 10 points: 1) the taxonomic identification, the life stage/size and gender of the test organisms; 2) collection methodology of the organisms, including collection location, conditions and methods; 3) holding and acclimation conditions in the laboratory; 4) exposure conditions such as test set up and exposure time, number of replicates and densities of organisms in tests and in test vessels; 5) range-finding test set up and schedule; 6) final test design, including details of controls and treatments, and replication options; 7) incubation conditions, specifying temperature, pH and oxygenation levels throughout the test; 8) test duration; 9) observations and endpoints; 10) test validity criteria and compliance. The recommendations were developed for the purpose of supporting future short-term ecotoxicological testing with stygofauna through providing consistency in the protocols. A discussion of the potential implications for groundwater managers and decision-makers committed to ERA for groundwater is included.

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.

Wells provide a distorted view of life in the aquifer: implications for sampling, monitoring and assessment of groundwater ecosystems.

When compared to surface ecosystems, groundwater sampling has unique constraints, including limited access to ecosystems through wells. In order to monitor groundwater, a detailed understanding of groundwater biota and what biological sampling of wells truly reflects, is paramount. This study aims to address this uncertainty, comparing the composition of biota in groundwater wells prior to and after purging, with samples collected prior to purging reflecting a potentially artificial environment and samples collected after purging representing the surrounding aquifer. This study uses DNA community profiling (metabarcoding) of 16S rDNA and 18S rDNA, combined with traditional stygofauna sampling methods, to characterise groundwater biota from four catchments within eastern Australia. Aquifer waters were dominated by Archaea and bacteria (e.g. Nitrosopumilales) that are often associated with nitrification processes, and contained a greater proportion of bacteria (e.g. Anaerolineales) associated with fermenting processes compared to well waters. In contrast, unpurged wells contained greater proportions of pathogenic bacteria and bacteria often associated with denitrification processes. In terms of eukaryotes, the abundances of copepods, syncarids and oligochaetes and total abundances of stygofauna were greater in wells than aquifers. These findings highlight the need to consider sampling requirements when completing groundwater ecology surveys.