Biodiversity hotspots are routinely identified by grid‐based analyses, despite grids encompassing different habitats, thus hindering the potential to assess which habitat type accounts for the conservation priority assigned to a grid. In this study, we aimed at identifying the main hotspots for the conservation of the European stygobitic Crustacea Copepoda Harpacticoida at the groundwater habitat scale. A multi‐metric approach was used, based on six biodiversity indicators: species richness, endemicity, evolutionary origin, phylogenetic rarity, taxonomic distinctness, habitat specificity. The Hot Spot Analysis, based on the statistics Getis‐Ord Gi*, was used to compare the local to the global average values of each indicator to identify hotspots of conservation. The operational units used to perform the analyses were the groundwater habitat types, in order to gather all the possible patterns of spatial occupancy in terms of habitat variability. Eight biodiversity hotspots of stygobitic Crustacea Harpacticoida were highlighted: 1) the Pyrenees (Spain and France), 2) the Jura Massif (France), 3) the Alpine arc (France, Switzerland and Italy) embracing southward the River Po alluvial plain and the Slovenian External Dinarides, 4) the Central Apennines (Italy), 5) the Carpathian and Balkan mountains in Romania and at the boundary between western Bulgaria and north‐west Macedonia, 6) the Dinaric Alps (from Croatia to Albania), 7) the Sardinia Island, 8) an area in central‐northern Europe embracing Denmark, the Netherlands and Germany. The hotspots showed a clear spatial distribution in southern Europe where they were distributed predominantly south to the 45th parallel, in line to what reiteratively observed in previous studies. Many hotspots embraced more than one habitat type. The adoption of discrete groundwater habitat types as working spatial units rather than grids provided a higher resolution of where the stygobitic harpacticoid species effectively live, with the possibility of intervening more precisely to preserve them and their habitats.

The distribution patterns of stygobitic crustacean harpacticoids at the boundaries of three different groundwater habitat types in Europe were analysed through a GIS proximity analysis and fitted to exponential models. The results showed that the highest frequency of occurrences was recorded in aquifers in consolidated rocks, followed by the aquifers in unconsolidated sediments and, finally, by the practically non-aquiferous rocks. The majority of the stygobitic harpacticoid species were not able to disperse across the boundaries between two adjacent habitats, with 66% of the species occurring in a single habitat type. The species were not evenly distributed, and 35–69% of them occurred from 2 to 6 km to the boundaries, depending on the adjacent habitat types. The distribution patterns were shaped by features extrinsic to the species, such as the hydrogeological properties of the aquifers, and by species’ intrinsic characteristics such as the preference for a given habitat type and dispersal abilities. Most boundaries between adjacent habitat types resulted to be “breaches”, that is transmissive borders for stygobitic harpacticoids, while others were “impermeable walls”, that is absorptive borders. Our results suggest that conservation measures of groundwater harpacticoids should consider how species are distributed within the different groundwater habitat types and at their boundaries to ensure the preservation of species metapopulations within habitat patches and beyond them.

Groundwater invertebrate communities in karst landscapes are known to vary in response to multiple environmental factors. This study aims to explore the invertebrate assemblages’ composition of an Apennine karst system in Italy mainly described by the Rio Gamberale surface stream and the Stiffe Cave. The stream sinks into the carbonate rock and predominantly feeds the saturated karst into the cave. For a minor portion, groundwater flows from the epikarst and the perched aquifer within it. The spatial distribution of the species belonging to the selected target group of the Crustacea Copepoda between the surface stream and the groundwater habitats inside the cave highlighted a different response of surface-water species and obligate groundwater dwellers to the hydrogeological traits of the karst unit. Our results suggest that fast endorheic infiltration routes promoted the drift of epigean species from the surface to groundwater via the sinking stream while most of the obligate groundwater dwellers come from the perched aquifer in the epikarst from diffuse infiltration pathways.

Understanding trait selection factors is vital for decoding the processes shaping species’ assemblages. However, trait-based studies in freshwater crustacean copepod assemblages are scarce, especially in groundwater environments. We explored how environmental filtering influences functional traits in copepod assemblages across four freshwater habitats (an alluvial aquifer, a hyporheic zone, a stream benthic zone and a lake littoral) along a depth gradient. Each habitat had distinct environmental templates based on light, temperature and dissolved oxygen. We analysed 4898 individuals from 43 copepod species and examined 12 morphological and life history traits. The results revealed significant differences in copepod traits among habitats, notably in ovigerous female biomass, egg biomass and ovigerous female percentages. Furthermore, despite some statistical uncertainty, notable differences were also observed in the number of juveniles, male-to-female abundance ratios and overall biomass. No significant differences were observed in juvenile biomass, egg characteristics, body size dimorphism or juvenile-to-adult ratios among habitats. The trait variations offer insights into copepod-mediated ecosystem services, particularly carbon recycling. To gain a deeper understanding of copepod adaptations to environmental features and anthropogenic changes, future research should consider additional functional traits, such as locomotion and feeding habits.

Groundwater is a crucial resource for humans and the environment, but its global human demand currently exceeds available volumes by 3.5 times. Climate change is expected to exacerbate this situation by increasing the frequency of droughts along with human impacts on groundwater ecosystems. Despite prior research on the quantitative effects of climate change on groundwater, the direct impacts on groundwater biodiversity, especially obligate groundwater species, remain largely unexplored. Therefore, investigating the potential impacts of climate change, including groundwater temperature changes, is crucial for the survival of obligate groundwater species. This study aimed to determine the thermal niche breadth of the crustacean amphipod species Niphargus longicaudatus by using the chronic method. We found that N. longicaudatus has a wide thermal niche with a natural performance range of 7-9 °C, which corresponds to the thermal regime this species experiences within its distribution range in Italy. The observed range of preferred temperature (PT) was different from the mean annual temperature of the sites from which the species has been collected, challenging the idea that groundwater species are only adapted to narrow temperature ranges. Considering the significant threats of climate change to groundwater ecosystems, these findings provide crucial information for the conservation of obligate groundwater species, suggesting that some of them may be more resilient to temperature changes than previously thought. Understanding the fundamental thermal niche of these species can inform conservation efforts and management strategies to protect groundwater ecosystems and their communities.

Aliphatic chlorinated hydrocarbons, notably tetrachloroethylene (also known as perchloroethylene [PCE]), are persistent, mobile, and toxic (PMT) and/or very persistent, mobile, and toxic (vPMT) groundwater pollutants, often exceeding safe drinking water thresholds. The present study delves into the groundwater risk assessment of PCE with a novel focus on the sensitivity of stygobitic species—organisms uniquely adapted to groundwater environments. Through a comparative analysis of the subchronic effects of PCE on the locomotion behavior of two copepod species, the stygobitic Moraria sp. and the nonstygobitic Bryocamptus zschokkei, we highlighted the inadequacy of the current European predicted-no-effect concentration of PCE for groundwater ecosystems. Our findings indicate significant behavioral impairments in both species at a concentration (32 ng/L PCE) well below the threshold deemed safe, suggesting that the current European guidelines for groundwater risk assessment may not adequately protect the unique biodiversity of groundwater habitats. Importantly, B. zschokkei demonstrated sensitivity to PCE comparable to or greater than that of the target stygobitic species, suggesting its utility as a substitute species in groundwater risk assessment. The present study adds to the limited research on the ecotoxicological sensitivity of groundwater species to PMT/vPMT chemicals and highlights the need for refined groundwater risk-assessment methodologies that consider the susceptibilities of stygobitic species. Environ Toxicol Chem 2024;43:2515–2527. © 2024 The Author(s). Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.