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.

New species of Elaphoidella Chappuis, 1929 and Schizopera Sars, 1905 (Copepoda: Harpacticoida) from two caves in northeastern and southern Thailand.

Two new copepod species from two caves in northeastern and southern Thailand are described. Elaphoidella isana sp. nov. and Schizopera paktaii sp. nov. were collected from the unsaturated zone of freshwater pools fed only by dripping water. They are closely related to E. intermedia Chappuis, 1931 and S. validior Sars, 1909, respectively. Elaphoidella isana sp. nov. can be distinguished from its closest relative by the armature of P4 and P5, and by the ornamentation of caudal rami. Schizopera paktaii sp. nov. differs from its relative S. validior in the armature of P1, P5 and caudal rami.

Modelling the effects of multiple stressors on respiration and microbial biomass in the hyporheic zone using decision trees.

Integrity of freshwater surface- and groundwater ecosystems and their ecological and qualitative status greatly depends on ecological processes taking place in streambed sediments overgrown by biofilm, in the hyporheic zone (HZ). Little is known about the interactions and effects of multiple stressors on biologically driven processes in the HZ. In this study, machine learning (ML) tools were used to provide evidence-based information on how stressors and ecologically important environmental factors interact and drive ecological processes and microbial biomass. The ML technique of decision trees using the J48 algorithm was applied to build models from a data set of 342 samples collected over three seasons at 24 sites within the catchments of five gravel-bed rivers in north-central Slovenia. Catchment-scale land use data and reach-scale environmental features indicating the HZ morphology and physical and chemical characteristics of water were used as predictive variables, while respiration (R) and microbial respiratory electron transport system activity (ETSA) were used as response variables indicating ecological processes and total protein content (TPC) indicating microbial biomass. Separate models were built for two HZ depths: 5-15 cm and 20-40 cm. The models with R as a response variable have the highest predictive performance (67-89%) showing that R is a good indicator of complex environmental gradients. The ETSA and TPC models were less accurate (42-67%) but still provide valuable ecological information. The best model show that temperature when combined with selected water quality elements is an important predictor of R at depth of 5-15 cm. The ETSA and TPC models show the combined effects of temperature, catchment land use and selected water quality elements on both response variables. Overall, this study provides new knowledge on how ecological processes occurring in the HZ respond to catchment and reach-scale variables, and provides evidence-based information about complex interactions between temperature, catchment land use and water quality. These interactions are highly dependent on the selection of the response variable, i.e., each response variable is influenced by a specific combination of predictive environmental variables.

A new blind groundwater-dwelling genus of the Cladocera (Crustacea: Branchiopoda) from the Korean Peninsula.

Groundwater-dwelling Cladocera are to date known only from the caves and hyporheic zones of the rivers in Europe (Belgium, England, France, Bosnia and Herzegovina, Macedonia, Slovenia, Spain), Turkey (European part) and Arabian Peninsula (Yemen). All these animals belong to the subfamily Aloninae Dybowski & Grochowski of the family Chydoridae Dybowski & Grochowski (Cladocera: Anomopoda). Examination of some samples collected by our colleagues - experts in Copepoda, Bathynellidae and Amphipoda-in South Korea led us to discovery of a new stygobiotic alonine genus, which is also the first record of a groundwater-dwelling cladoceran in Asia. Aims of this paper are: (1) to describe Korealona gen. nov., represented by two species, Korealona karanovici gen. nov., sp. nov. found in three different localities, and K. choi sp. nov., also found in three different localities; and (2) to discuss its differences from other genera of this subfamily and possible phylogenetic position of this genus among the aloninse. Distribution ranges of two species of Korealona gen. nov. are not unequivocally associated with present-day main river basins in Korea, but more sampling efforts are needed for final conclusions on their distribution ranges in Korea and closest countries and understanding of the genus evolutionary history.

Amended diagnosis of the genus Nannodiaptomus (Copepoda, Calanoida), based on redescription of N. phongnhaensis and description of a new species from caves in central Vietnam.

Nannodiaptomus phongnhaensis Dang & Ho, 2001, an endemic stygobiotic diaptomid from Vietnam and the type species of the genus Nannodiaptomus Dang & Ho, 2001, is redescribed on the basis of a new material from its type locality: Hang Phong Nha Cave, Quang Binh province, central Vietnam. We also designated a neotype, because the type material has been lost. The diagnosis of the genus is amended, based on the description of the neotype. The taxonomic position of the genus in the subfamily Speodiaptominae is proposed, and its relationships with other genera of this subfamily are discussed. A new cave-dwelling species Nannodiaptomus haii sp. nov., from a cave near the type locality of N. phongnhaensis, is also described and illustrated. The new species differs from the type species of the genus, among other things, by the structure of male's left swimming leg 5, where the inner margin of the apical process and the sub-terminal seta on the swimming leg 5 Exp-2 have small serrated membranes, but these bear 3-4 distinct denticular protrusions in N. phongnhaensis.

Two new stygobiotic copepod species from the Tibesti area (Northern Chad) and a re-description of Pilocamptus schroederi (van Douwe, 1915).

Between 4th and 18th March 2014, an international group of biologists carried out a hydrobiological expedition to the Ounianga-Tibesti area of northern Chad (Africa). The Tibesti is a desert volcanic area, intersected by the beds of ancient rivers which were active in the Tertiary. In deep canyons there are small water bodies (gueltas/aguelmans), fed by rain and spring water. They are rich in zooplankton, benthos, and even fish, but their groundwater fauna has previously been unknown. Groundwater samples collected in the vicinity of one guelta contained Syncarida, Isopoda, and Copepoda. Among the latter, two new species were recognised, Haplocyclops (H.) henrii sp. nov. and Parastenocaris joi sp. nov., together with a third species, Pilocamptus schroederi (van Douwe, 1915), previously known only from the littoral zone and wet mosses of Lake Victoria. The Tibesti area is thus the third known location of P. schroederi. All these species have a Gondwanaland distribution and are probably relicts of the Tertiary fauna, formerly widespread in the Sahara. Together with descriptions of two new species, a detailed re-description of P. schroederi is presented, along with remarks on their ecology.

Effects of increased temperature on metabolic activity and oxidative stress in the first life stages of marble trout (Salmo marmoratus).

Climate change may result in future alterations in thermal regime which could markedly affect the early developmental stages of cold water fish due to their expected high sensitivity to increasing temperature. In the present study, the effect of temperature increase of 2, 4 and 6°C on the oxygen consumption rate (R), the activity of respiratory electron transport system (ETS) and oxidative stress have been studied in four developmental stages of the marble trout (Salmo marmoratus)-eyed eggs, yolk-sac larvae and juveniles of 1 and 3 months. Oxygen consumption rate and ETS activity increased with level of development and with temperature in all four stages. ETS/R ratios decreased during development and correlated with temperature in eyed eggs, larvae and juveniles of 1 month, but not in juveniles of 3 months. Low ETS/R ratios at higher temperatures indicate stress response in eyed eggs, the most temperature sensitive developmental stage. Catalase (CAT) and glutathione reductase (GR) activities increased during development, but responded differently to elevated temperature in the different developmental stages. Stress in eyed eggs, caused by higher temperatures, resulted in increased oxygen consumption rate and increased activities of CAT and GR. Larvae were sensitive to increased temperature only at the highest experimental temperature of 16°C. Increased temperature did not stress the metabolism of the juveniles, since they were able to compensate their metabolic activity. The earlier developmental stages of marble trout are thus more sensitive to temperature increase than juveniles and therefore more endangered by higher water temperatures. This is the first report connecting oxygen consumption, ETS activity and ETS/R ratio with the activities of antioxidant enzymes in relation to increased temperature in salmonids.

Two new stygobiotic species of Elaphoidella (Crustacea: Copepoda: Harpacticoida) with comments on geographical distribution and ecology of harpacticoids from caves in Thailand.

Elaphoidella thailandensis sp. nov. and E. jaesornensis sp. nov., collected during an investigation of cave-dwelling copepod fauna in the northern part of Thailand, are described and figured herein. The new species were collected from pools filled by percolating water from the unsaturated zone of a karstic aquifer in Phitsanulok and Lampang Provinces, respectively. Elaphoidella thailandensis, from Tham Khun cave, is distinguished from its congeners by the two-segmented endopod of pediger 1, the absence of endopod on pediger 4, and the setal formula 4, 5, 6 for the distal exopodal segment of pedigers 2-4. Elaphoidella jaesornensis, from Tham Phar Ngam cave, is distinguished from its most closely related species, E. namnaoensis Brancelj, Watiroyram & Sanoamuang, 2010, by the armature formula of the endopod of pedigers 2-5. The geographical distribution and ecology of Harpacticoida from Thai caves is also presented.

Groundwater drift as a tracer for identifying sources of spring discharge.

Groundwater invertebrate drift, collected from the spring outlets at the interface of vadose and phreatic zones, has been examined for its potential for identifying sources of discharge from a karst aquifer. Concurrently, major ion geochemistry, dissolved inorganic carbon (δ13CDIC), particulate organic carbon (δ13CPOC), and naturally occurring stable isotopes of oxygen and tritium (δ18O, 3H) were investigated over a period of 1 year in two outlets, a temporary (TS) and a perennial (PS) spring. A few differences in major ion geochemistry and stable isotope composition were found between the two springs together with moderate seasonal variability. In contrast, invertebrate drift showed clear differences between TS and PS springs in density and composition. Canonical correspondence analysis showed the presence of two distinct groups of samples from TS and PS, with Ca2+ as the only significant explanatory variable for differences in drift composition. Finally, certain species from the drift were found to be useful tracers for distinguishing between the phreatic and the epikarst and vadose zones as the origin of spring water.

The thermal tolerance of crayfish could be estimated from respiratory electron transport system activity.

Whether electron transport system (ETS) activity could be used as an estimator of crayfish thermal tolerance has been investigated experimentally. Food consumption rate, respiration rates in the air and water, the difference between energy consumption and respiration costs at a given temperature ('potential growth scope', PGS), and ETS activity of Orconectes limosus and Pacifastacus leniusculus were determined over a temperature range of 5-30°C. All concerned parameters were found to be temperature dependent. The significant correlation between ETS activity and PGS indicates that they respond similarly to temperature change. The regression analysis of ETS activity as an estimator of thermal tolerance at the mitochondrial level and PGS as an indicator of thermal tolerance at the organismic level showed the shift of optimum temperature ranges of ETS activity to the right for 2° in O. limosus and for 3° in P. leniusculus. Thus, lower estimated temperature optima and temperatures of optimum ranges of PGS compared to ETS activity could indicate higher thermal sensitivity at the organismic level than at a lower level of complexity (i.e. at the mitochondrial level). The response of ETS activity to temperature change, especially at lower and higher temperatures, indicates differences in the characteristics of the ETSs in O. limosus and P. leniusculus. O. limosus is less sensitive to high temperature. The significant correlation between PGS and ETS activity supports our assumption that ETS activity could be used for the rapid estimation of thermal tolerance in crayfish species.

A new method for early assessment of effects of exposing two non-target crustacean species, Asellus aquaticus and Gammarus fossarum, to pesticides, a laboratory study.

A reliable method is needed for assessing the condition of aquatic animals and their resistance to toxic pollutants. The physiological responses of two freshwater crustaceans, Asellus aquaticus and Gammarus fossarum, following in vitro exposure to two pesticides (atrazine and imidacloprid), were measured by a combination of electron transport system (ETS) activity and respiration (R). Short-term exposure concentrations were selected according to standard toxicity tests and ranged from 0.01 mg L(-1) to 10 mg L(-1). When pesticide concentration was greater than 1 mg l(- 1) (which is below the LC(50) [48 hours] determined for both species), A. aquaticus and G. fossarum responded to short-term exposure with elevated levels of R and/or lower levels of ETS activity. One hour exposure to concentrations of up to 10 mg L(-1) showed an effect in both test species. Laboratory tests confirmed that G. fossarum is more sensitive to short-term pesticide exposure than A. aquaticus. The combination of these two methods provides a useful and effective tool for assessing the general condition of aquatic animals. It also enables to determine toxic effects on freshwater biota of specific or combined pollutants. ETS/R ratio may be used as a quick predictor of effects on organisms exposed to pesticides and other stress factors such as changes in temperature, light, salinity, oxygen concentration and food.

Effects of exposing two non-target crustacean species, Asellus aquaticus L., and Gammarus fossarum Koch., to atrazine and imidacloprid.

The physiological responses of two freshwater crustaceans, Asellus aquaticus L. and Gammarus fossarum Koch., following in vitro exposure to two pesticides were measured. Both species responded to short-term exposure with elevated levels of Respiration and/or lower levels of Electron Transport System (ETS) activity. 1 h exposure to concentrations of up to 10 mg L(-1) showed an effect in both test species. Laboratory tests confirmed that G. fossarum is more sensitive to short-term pesticide exposure than A. aquaticus. ETS/R ratio may be used as a quick predictor of effects on organisms exposed to pesticides.