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Groundwater Ecology Summary Report

REPORT ON GROUNDWATER ECOLOGY

June 2017

v. 1

Authors:

Professor Anne Robertson. Department of Life Sciences, Roehampton University.

Dr Christian Griebler. Helmholtz Zentrum München.

Dr Florian Malard. Université Lyon 1/ CNRS

Professor Diana Galassi, University of L’Aquila

Associate Professor Grant Hose, Department of Biological Sciences, Macquarie University

Summary report of the special session at the WFD CIS Groundwater Working Group meeting held in Malta on 25-56 April

2017.

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1 OverviewPromoting a common implementation strategy for the assessment of groundwater ecosystem health and risk.

1a. Introduction

Groundwater comprises 94% of liquid freshwater on earth and, from a human perspective, is a vital resource that provides, among other things, water for drinking and irrigation. It is also essential to support surface water bodies, or terrestrial systems such as wetlands. From an ecosystem perspective, however, groundwater is living space for organisms and reactant in biogeochemical processes. Groundwater ecosystems contain unique food webs, simple in comparison to surface waters because most groundwaters contain low levels of carbon (food) and there is insufficient energy to sustain more than three to four trophic levels.

Microorganisms and viruses are ubiquitous in groundwater ecosystems and occur down to depths of several thousand meters. The cycling of carbon and nutrients, as well as other elements, in aquifers are mediated by these microbial communities, making them essential players in ecosystem services such as water purification including biodegradation of contaminants and elimination of pathogens. As well as the microbes, meio- and macrofauna species occur in groundwater. Many such species are found in groundwater only and have restricted distributions; thus, they make a unique contribution to global biodiversity. Groundwater fauna are mostly invertebrates (crustaceans, molluscs and worms) and inhabit pore spaces and fractures in the aquifer. Fish and amphibians are rare and are limited to karst aquifers.

Figure 1: Typical representatives of microbes and invertebrates in groundwater ecosystems. Left: Bacteria; Middle: Amoeba and heterotrophic flagellates; Right: Crustaceans (Isopoda, Amphipoda, Syncarida), Turbellarian flatworm and snail. The arrows indicate the flow of energy and food web relationships.

Groundwater animals are adapted to the low energy conditions; they reproduce slowly and their population sizes are small. These characteristics, together with their restricted distributions make groundwater fauna vulnerable to the pressures that occur in groundwaters, which include contamination, alteration of temperature due to extraction and storage of heat and cold, and overexploitation.

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In this report we detail recent groundwater ecological research that demonstrates the contribution of these unique ecosystems to Natural Capital and the ecosystem services that they deliver. We argue that, as a result of these services, groundwater ecosystems should be treated in a similar way to surface waters and feature strongly in any revision of the Groundwater Directive. The research we report here shows how these ecosystems can be characterised and risk assessed. At the present time, groundwater ecosystems are not routinely monitored; we outline a simple toolbox containing a variety of measures that can be used to monitor the health of groundwater ecosystems. Finally we identify some remaining knowledge gaps and suggest ways in which these can be filled.

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2 Discussion

2a. Recent developments, timeline of developments in the field

From early descriptive work on identity and distributions of individual stygobionts (obligate groundwater organisms) in the 1700’s, groundwater ecology has developed into a mature re-search field making major contributions to important ecological concepts (e.g. ecotone concept in the 1990’s) and to knowledge about distributions, evolution and the functional role of biodiversity (in the 2000s) (Danielopol & Griebler 2008). More recently the research focus of groundwater ecology has moved to identifying the delivery of ecosystem services (following the MEA 2005) and to how these ecosystems can be assessed (Griebler et al. 2014; Griebler & Avramov 2015).

2b. Current state of knowledge, results, findings and ongoing research, including coordination across Europe and globally

CURRENT STATE OF KNOWLEDGE, AVAILABLE RESULTS AND FINDINGS: Groundwater ecosystems make an important contribution to global Natural Capital (the world’s stocks of natural assets such as geology, soil, air, water and all living things (World Forum on Natural Capital 2016)). There are more than 2000 species of stygobiont taxa described for Europe, with many more awaiting description (Ferreira 2005). The number of bacterial and protozoan species exceeds that of metazoan by several orders of magnitude (Griebler & Lueders 2009). Our research shows that the natural capital of groundwater organisms results in the delivery of a wide range of ecosystem services. Among the most recognized services is the natural purification of water (Griebler & Avramov 2015). Provision of clean water for drinking and other purposes is of enormous economic value. Numerous studies have underlined the essential role of mi-crobial communities in biodegradation of contaminants (e.g. Meckenstock et al. 2015). Micro-bial communities also mediate the attenuation of nitrate and ammonia through denitrification and nitrification (Rivett et al. 2008) in the many nitrate contaminated sites across Europe and contribute to the inactivation and elimination of pathogens (e.g. Deng et al. 2014). In sum-mary, these ecosystem services help to save billions of Euros otherwise required for ad-vanced water treatment.

Higher trophic levels, represented by metazoan taxa and protozoa, enhance the ability of mi-crobial communities to perform these ecosystem services. The construction of tubes and bur-rows and the settling of faeces by metazoan taxa increase nutrient availability and hence activity of bacteria (Mermillod-Blondin 2011). Recently, Weitowitz et al. (unpubl.) showed that groundwater macrofauna significantly increased protozoan abundance, and Peralta et al. (un-publ.) found that breakdown of the pharmaceutical ibuprofen (pain killer) by groundwater bac-teria was significantly greater when protozoan grazers were present, providing a clear link between the larger invertebrates and ecosystem services by microbes within the groundwater environment.

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Apart from their value in ecosystem service delivery, there is great value in considering eco-logy in groundwater assessment. Physico-chemical analysis provides a snapshot of environ-mental conditions at a single point in time and can only cover a selected number of paramet -ers. Biological analysis can provide a time-integrated picture with indirect detection of un-known threats. Biological analyses can also be used as early warning systems (e.g. in an on-line biomonitor; Gerhardt et al. 2003) and are sensitive where chemical analyses fail (e.g. im-pact of heat) (Griebler et al. 2016). Last, but not least, biological parameters are extremely useful subsequent to an impact because they can help to document the return to good condi -tion (Griebler et al. 2010). For example, Mauffret et al. (2017) showed that groundwater mi-crobial communities hold the imprints of an exposure to atrazine herbicide more than 10 years after its withdrawal.

The characterisation of GW ecosystems is an important foundation for the inclusion of GW ecosystem assessment and biomonitoring schemes in national and European legislation. Methods for characterizing groundwater habitats and their associated fauna are well-ad-vanced and have been trialled at various scales. There is an emerging consensus on the eco-hydrogeological criteria for habitat typology (9 convergent proposals since 2009); these are pore space, permeability and groundwater recharge mode. Cornu et al (2013) have de-veloped a groundwater habitat map of Europe using these criteria at the EU level (Fig. 2; left). Weitowitz et al. (2017) refined this EU map at a national scale. They identified twelve geo-habitats in the UK based on transmissivity, porosity and chemistry. They then developed a score for geo-habitat quality using critical thresholds for each criteria and also cave develop-ment and habitat space and used this to map groundwater habitat quality across the UK.

Figure 2: There are many European-wide data sets for characterizing groundwater ecosystems in-cluding the groundwater habitat map of Europe (Cornu et al. 2013) on the left, the European ground-water crustacean data set (Zagmajster et al. 2014; Eme et al. 2015) in the middle, and the groundwa-ter bioregional map of Europe (Stoch et al. in progress) on the right.

There is already a comprehensive European groundwater crustacean data set (Zagmajster et al. 2014) collected from individual records and initiatives at the national (e.g. Johns et al. 2014) and European level (e.g. PASCALIS (EU FP5 2002-2004), BIOFRESH (EU FP7 2009-2014). Using this data set, biological reference values can be determined in terms of species richness for groundwater bodies of Europe using the relationship between regional and local

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species richness (Malard et al. 2009). Groundwater community composition has also been determined for European Bioregions and further refined at the national scale in selected coun-tries (e.g. France (Ferreira et al. 2005), Germany (Stein et al. 2012), Italy (Stoch & Galassi 2010). Thus, there already exists a clear habitat typology with habitat specific biological refer-ences for groundwater ecosystems at the EU level with some national level refinements.

With respect to microbes and viruses, scientists have a clear picture of what to expect in healthy or only moderately impacted groundwater ecosystems in terms of numbers, biomass and activity. However, Europe wide explorations are so far missing. Recently, strong efforts have been made in Germany (projects funded by the Federal Environmental Agency (UBA) and the Ministry of Education and Research (BMBF) such as GroundCare), in Switzerland (organized and funded by the Federal Office for the Environment (FOEN)), and outside Europe in Australia (see below). Since groundwaters differ from surface waters and contamin-ated waters because they are oligotrophic with low energy, the microbial communities of groundwater can be characterised, as a first step, using simple measures: biomass (total prokaryotic cell counts), activity (ATP, intact cells), and energy (ready assimilable organic car-bon) (Fig. 3; Griebler et al., in prep.).

Figure 3: Groundwater microbial communities are typically adapted to life at low energy and comprise small prokaryotic cells (bacteria) with low abundance and activity under undisturbed conditions. The left chart shows the separation of groundwater samples from surface water samples by total bacterial cell counts (TCC, in X-axis) and ATP (Y-axis) (data from Hammes 2011 and Griebler et al., unpubl. data). Since microbial communities are very responsive to an increase in organic matter input, easy-to-measure and cheap parameters such as total cell counts and activity provide excellent indicators of organic pollution. The right chart shows TCC and ATP measurements in samples from undisturbed to disturbed (organically contaminated) groundwater, as well as its recovery (Griebler et al. unpubl. data).

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Total Cell counts [Cell-

Total Cell counts [Cell-


Biological assessment of groundwaters is being incorporated (along with traditional water quality and quantity measures) into revisions of water management plans in New South Wales, Australia where the goal is to ensure that environmental objectives of those plans are being met. A tiered framework for assessing groundwater health is already in use (Korbel & Hose 2011). In this framework, the first tier provides a preliminary assessment of groundwater ecosystem health (pass or fail) that can be done in a single visit with benchmarks derived from the literature. If a site fails this first tier of assessment, it undergoes the second tier of assessment which uses benchmarks derived from field sampling (e.g. % Crustacea present, % Oligochaeta, stygoxene: stygobite ratio, concentrations of DOC, nitrate and synthetic chemicals). Extensive testing indicates that Tier 2 benchmarks reliably distinguish between healthy and impacted ecosystems, and the flexibility of the benchmarks used enables it to be adapted to most situations and operational budgets. This assessment model has also been used successfully in Germany.

Toxicity data for groundwater taxa are mainly from acute tests. Although some groundwater species have proved to be more sensitive to some pollutants than their close epigean relat -ives (Caschetto et al., 2017; Di Lorenzo et al., 2015 a-c), likely due to their lower metabolic rates (Di Lorenzo et al., 2015d), other studies have obtained different results (Avramov et al., 2013; Reboleira et al., 2013). Currently, an online-biomonitor (Gerhardt et al. 2003) has been adapted for use with groundwater metazoans. It can either be used as a flow-through early warning system (similar to a Daphnia test) or for acute and chronic ecotoxicological sub-stance evaluation. This, for the first time, allows analysis under groundwater-specific condi-tions (see GroundCare project, https://bmbf.nawam-rewam.de/projekt/groundcare/; LimCo International GmbH, https://www.limco-int.com/).

Estimating the risk that groundwater contamination may pose to stygobionts using epigean taxa as surrogates, is being pioneered for groundwater bodies in the LIFE AQUALIFE project granted by the European Community in the Abruzzo region of Italy. This biological risk as-sessment approach examines both exposures resulting from occurrence of chemicals and the effects of such emissions on the structure and functions of the metazoan community. The main purpose is to develop a procedure for groundwater ecosystem risk assessment (AQUALIFE GRA) that integrates the ‘bio-risk’ into the WFD – Groundwater Risk Assessment concept (GRA WFD). The Bio-risk is the sum of risk quotients (RQs) that are calculated as the ratio of the measured maximum environmental concentration of a given compound to its pre-dicted no effect concentration (PNEC). The PNEC values are usually determined on the basis of results from single species laboratory tests or, in a few cases, from model ecosystem tests, taking into account adequate assessment factors. The first version of AQUALIFE GRA has recently been applied to 27 groundwater bodies in southern Italy and demonstrated that the bio-approach enhances the level of information on the groundwater bodies at risk of failing the quality objectives of the article 4 of the WFD. Biological GRA is applied once in a River Basin Management Plan cycle and can be done by local Environmental Agencies (a week long training course is sufficient). Here, AQUALIFE will provide a user-friendly software package.

Tool-Box for the biological and ecological characterisation of groundwater bodies:

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https://www.limco-int.com/

https://bmbf.nawam-rewam.de/projekt/groundcare/


The authors of this report (Galassi, Griebler, Hose, Malard, Robertson) highly recommend that the preliminary toolbox for the biological and ecological characterisation and assessment of groundwater bodies is applied and further developed. This toolbox is underpinned by the groundwater ecosystem research outlined in this report that followed the call for more re-search in Recital 20 of the EU-GWD ten years ago. The toolbox includes measures that are quick to undertake, that can be implemented by non-specialists and that can be easily incor-porated into existing monitoring programmes for the assessment of groundwater chemical and quantitative status (Fig. 4).

Figure 4: The preliminary toolbox. The lower grey box highlights simple measurements, such as TCC vs. ATP (microbial level) and stygobionts vs. stygoxenes (faunal level) that can already be applied for the assessment of groundwater ecosystem status.

Detailed development of the aims and objectives of the toolbox and the protocols therein are in progress, however, measures such as the high-throughput measurement of total bacterial numbers and their activity (Fig. 3), as well as the number of stygobionts and their ratio to fauna coming from surface ecosystems can readily be applied (Fig. 4).

RESEARCH CO-ORDINATION AND STRUCTURE: In the last 15 years there have been two integrat-ive European level projects relevant to groundwater ecosystems – PASCALIS (2002-2004) and BIOFRESH (2009-2014). In FP 7, the ITN GOODWATER (2008-2012) dealt with biolo-gical attenuation of organic contaminants in aquifers. Currently, there are two Horizon 2020 ITN projects, HYPOTRAIN and INTERFACES, focussing on biogeochemical processes in the hyporheic zone, an important ecotone between rivers and groundwaters. However, most re-search on groundwater ecosystems is currently funded at the national level and so co-ordina-tion at the Europe scale is minimal apart from informal researcher contacts. This leads to frag-mentation of the research effort. Most of the research presented in this report has been un-

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dertaken in the groups led by the authors of this report. However, there are many other insti -tutions and colleagues that are active in Europe and internationally.

2c. Remaining challenges, required future research, identified potential limiting factors and stumbling blocks

Great advances have been made recently in understanding groundwater ecosystems, both in terms of their importance in ecosystem service delivery and also the ways they could be in-cluded in GW assessment and biomonitoring as part of the WFD and Groundwater Directive.

However, some challenges remain which are briefly highlighted below.

(1) While groundwater fauna have been studied in many European areas, there are still gaps in groundwater fauna distribution maps where we totally lack information. We now need to focus our efforts to fill these gaps. This is particularly true for groundwater microbial communities. Although microbes are directly involved in the most essential ecosystem services, with the exception of some areas of Germany, we have no spa-tially inclusive and comprehensive data. The toolbox necessary to collect this data has already been developed (Fig. 4) and we now need to apply it across Europe.

(2) A characterisation of groundwater according to its ecosystem character is a required basis for any kind of ecosystem health assessment and biomonitoring. As highlighted in Figure 2, characterisation schemes for groundwater habitats have been successfully developed. Moreover, for individual countries (e.g. France, Germany, Italy, UK), these have already been refined and related to groundwater fauna distribution. The different approaches now need to be harmonized and applied to other countries in Europe as well.

(3) As presented at the CIS working Group Meeting in San Lawrenz, Malta, a first tool box for the assessment of groundwater ecosystems including faunal and microbiological criteria (including an assessment index) is available (Fig. 4). Furthermore, we can show that these criteria are sensitive to the impact of surface water, organic pollution (Fig. 3) and temperature impacts. Testing for criteria that allow us to trace contamina-tion with micropollutants and nutrients (e.g. nitrate) is still ongoing.

(4) The first groundwater ecosystem status index (Korbel & Hose 2011) has now been tested at sites in Australia and Germany, but urgently awaits further evaluation and validation with respect to different groundwater pressures within European countries.

(5) As mentioned above, the WFD groundwater risk assessment (GRA) approach can easily be extended to include the assessment of the biological/ecological risk. A new approach developed in the EU LIFE project AQUALIFE allows assessing the biological risk to groundwater bodies based on GRA data and matching them with ecotoxicolo-gical data available for groundwater fauna. This easy and low-cost approach now awaits broad application and validation across Europe.

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There are 3 major inter-linked overarching limitations that govern the ability of the research community to sustainably address the challenges highlighted above. These are:

The absence of clear acknowledgement of the presence, and importance, of groundwater ecosystems in the WFD, the GWD, and national laws.

The lack of Europe scale co-ordination of the research efforts The lack of Europe level funding.

Rewording the recitals of EU-WFD and EU-Groundwater Directive (GWD) will help to address the first gap by acknowledging the presence and importance of groundwater ecosystems among member states. The Groundwater Directive contains an ‘ecosystem perspective’ for groundwater (Recital 20) but this is completely absent in the WFD. We propose that several paragraphs in the Groundwater Directive recitals should be extended to include consideration of the chemical and ecological status (e.g. recital 6, 7, 8). We also propose that the text of Recital 20 should be modified to ‘Coordinated efforts among member states should be con-ducted in order to elaborate a common implementation strategy for assessing the health of groundwater ecosystems and an assessment scheme for the determination of the ecological status of subterranean water bodies to complement the ones already in place for the quantit -ative and chemical status. In preparation for this, the following components should be de-veloped: i) a characterisation scheme for groundwater ecosystems, ii) a common approach/conceptual methodology for the monitoring of groundwater ecosystems and the identification of the ones being at risk and iii) criteria for the assessment of the ecological status / ground-water ecosystem health.

In the national water laws of some individual EU member states and associated countries, groundwaters are already defined as water bodies that contribute to the natural capital of freshwaters (together with surface and coastal waters) and that deserve protection as a re-source but also as a habitat of diverse communities (e.g. Federal Water Act of Germany, Wa-ter Ordinance of Switzerland). However, most groundwater-specific regulations lack these aspects. In the authors’ opinion, it is only a matter of time before the unequal treatment of sur-face and subsurface waters is challenged by environmental associations and NGOs in the courts.

A Europe wide coordination of research and application of groundwater ecosystem health is-sues can only be achieved with support of the EU. The authors propose the submission of a proposal for a COST action at the earliest possible time to actively foster a European ground-water ecosystem network. We also recommend that the research topic of groundwater eco-systems and their assessment is included in the list of supported topics put forward by the CIS Working Group Groundwater to the DG Research for consideration in the new framework programme.

Support for an EU research call on groundwater ecosystems assessment, initiated and sup-ported from the CIS Working Group Groundwater, specifically to address the second and third

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gaps will help provide Europe wide cohesion in the research effort and much needed funding. Emphasis is on translating knowledge on groundwater ecosystems accumulated over the last 10 years into a common implementation strategy for assessing the health of groundwater ecosystems by defining a tool-box of practical tools for end-users.

2d. Analogies and differences in ecological perspectives between surface water and groundwa-ter in research

The WFD defines ecological status as the expression of the quality of the structure and func-tioning of aquatic ecosystems associated with surface waters, classified in accordance with quality elements provided in Annex V. The biological quality element designated as “composi-tion and abundance of benthic invertebrate fauna” in Annex V of the WFD can also be applied to groundwater metazoans and is readily available for many groundwater bodies across Europe (see section 2b).

As defined in Annex II of the WFD, type-specific biological reference conditions for inverteb-rate species richness and community composition can also be determined for groundwater bodies under undisturbed conditions based on available knowledge (see section 2b). These type-specific biological reference conditions can further be used to assess whether the inver-tebrate community of a groundwater body significantly deviates from reference conditions, thereby adding a community-based assessment of groundwater bodies to the assessment of good groundwater chemical and quantitative status.

Groundwaters differ from surface waters in that they do not support photosynthetic microor-ganisms. So, in addition to the invertebrates and instead of algae and other common indicat-ors used for surface waters, heterotrophic bacteria that are adapted to life with low energy conditions and that are very responsive to changes in the aquifers (see section 2b) must be considered in assessing groundwater ecosystem health. In Germany and Switzerland meas-urement of key parameters pertaining to groundwater communities has complemented the assessment of chemical status of groundwater bodies (see Figure 3).

2e. Experience with operationalising groundwater ecology monitoring and cooperation with authorities

In the absence of a common implementation strategy for assessing the health of groundwater ecosystems, the use of groundwater organisms in groundwater monitoring is carried out mainly at regional level on a case-by-case basis. There are however numerous successful applications of groundwater biomonitoring including the use of crustaceans as sentinel organ-isms to evaluate groundwater quality (Stein et al. 2010; Marmonier et al. 2013; Di Lorenzo & Galassi 2013; Di Lorenzo et al. 2014; Di Marzio et al. 2013), the use of microbial communities for assessing surface water inputs of organic matter and nutrients to groundwater (Steube et al. 2009; Griebler et al. 2010; Stein et al. 2010; Mermillod-Blondin et al. 2013), and the coup -

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ling of groundwater modelling and biological indicators for identifying river/aquifer exchanges (Graillot et al. 2014).

In Germany, the assessment of groundwater ecosystem health is tackled in the framework of the GroundCare project in close cooperation with stakeholders (water works) and authorities (Bavarian Environment Agency).

In Italy, a study of the invertebrate composition in the hyporheic zone of one of the largest basins in the country was funded by the Basin Authority of the River Adige in cooperation with the APAT Trento (the local authority in charge of the chemical characterization of surface wa-ters and hyporheic waters of this basin).

In Australia, groundwater monitoring on cotton farms has been possible with sampling under-taken by farmers, with the support of scientists for subsequent analysis and interpretation. The engagement of the farming community with this trial program has been instrumental to achieving cost effective monitoring and effective education and knowledge dissemination.

3 Summary and conclusionThe study of groundwater ecosystems, a mature research field of water sciences, is revealing the diversity of groundwater organisms, including metazoan taxa, protozoa and bacteria as well as their role in water purification and thus the provision of clean drinking water. As a res-ult of the many ecosystem services mediated by groundwater organisms, there is a high ad-ded value in adapting groundwater environmental policies and monitoring practices so that they integrate life in groundwater as an essential component of groundwater resources. There is also ample evidence that groundwater organisms reflect the effects of human activities at community to gene levels making them excellent indicators of groundwater pollution and de-terioration. Notably, groundwater organisms can be used to assess the recovery of groundwa-ter ecosystems following cessation of pollution.

In addition to a series of 5 presentations at the 25-26 April 2017 meeting of the CIS Working Group Groundwater in San Lawrenz, Malta, we provide in this summary report a snapshot of the state of the art knowledge in i) characterizing groundwater habitats and their associated metazoans and micro-organisms; ii) selecting appropriate ecological criteria for evaluating groundwater ecosystem health and; iii) elaborating methods for assessing groundwater com-munities at risk of failing to deliver ecosystem services. We highlight considerable advances made in these three aspects since the 2006/2007 iteration of the groundwater directive, not-withstanding the fact that a number of well-identified challenges remain to be addressed.

We then identify three major inter-linked gaps that limit the ability of the research community to address these challenges, namely, the lack of clear acknowledgement of the presence, and importance, of GW ecosystems in the GW Directive, the lack of Europe scale co-ordination of the research effort and the lack of Europe level funding to address specifically these chal-lenges. We outline a number of solutions for translating the knowledge on groundwater eco-

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systems accumulated over the last 10 years into a common implementation strategy for as-sessing the health of groundwater ecosystems and identifying those at risk of failing to deliver important ecosystem services.

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5 Contact informationFor potential clarifying questions, please provide information on your and your key partner's contact details, in -cluding name, organisation, email and telephone number. This information will be treated confidentially.

Professor Anne Robertson. Department of Life Sciences, Roehampton University. [email protected], +44 2083923456

Dr Christian Griebler. Helmholtz Zentrum München. [email protected] , +49 8931872564

Dr Florian Malard. Université Lyon 1/ [email protected] , +33 0472431561

Professor Diana Galassi. University of L’[email protected] , +39 0862433227

Dr Tiziana Di Lorenzo. ISE-CNR, Florence [email protected], +39 390555225918

Associate Professor Dr Grant Hose. Department of Biological Sciences, Macquarie [email protected] , +61 298506296

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