Under global change, the ion concentration of aquatic ecosystems is changing worldwide. Many freshwater ecosystems are being salinized by anthropogenic salt inputs, whereas many naturally saline ones are being diluted by agricultural drainages. This occurs concomitantly with changes in other stressors, which can result in additive, antagonistic or synergistic effects on organisms. We reviewed experimental studies that manipulated salinity and other abiotic stressors, on inland and transitional aquatic habitats, to (i) synthesize their main effects on organisms' performance, (ii) quantify the frequency of joint effect types across studies and (iii) determine the overall individual and joint effects and their variation among salinity–stressor pairs and organism groups using meta-analyses. Additive effects were slightly more frequent (54%) than non-additive ones (46%) across all the studies ( n = 105 responses). However, antagonistic effects were dominant for the stressor pair salinity and toxicants (44%, n = 43), transitional habitats (48%, n = 31) and vertebrates (71%, n = 21). Meta-analyses showed detrimental additive joint effects of salinity and other stressors on organism performance and a greater individual impact of salinity than the other stressors. These results were consistent across stressor pairs and organism types. These findings suggest that strategies to mitigate multiple stressor impacts on aquatic ecosystems should prioritize restoring natural salinity concentrations. This article is part of the theme issue ‘Salt in freshwaters: causes, ecological consequences and future prospects’.

One of the main challenges in ecology, biogeography and evolution is to understand and predict how species may respond to environmental changes. Here we focus on the deep subterranean environment, a system that minimizes most of the typical uncertainties of studies on epigean (surface) environments. Caves are relatively homogeneous habitats with nearly constant environmental conditions and simplified biological communities, allowing to control for biotic interactions. Thus, this particular system could be considered a natural habitat whose environmental conditions are similar to what can be reproduced in a laboratory, being an ideal model system for ecological, biogeographical and evolutionary studies. Subterranean species may potentially be used to assess the capability to persist in situ in a global change scenario, as they cannot accommodate to drastic changing conditions by behavioural plasticity, microhabitat use or by migrating to distant, more suitable areas, something frequent in epigean environments. In order to provide accurate predictions of the response of the subterranean biodiversity to climate change, we encourage evolutionary biologist, biogeographers and conservation biologist to work in this interesting ecosystem

Physiological traits are key in determining the vulnerability of narrow range, highly specialized animals to climate change. It is generally predicted that species from more stable environments possess lower thermal tolerance breadths and thermal plasticity than those from more variable habitats – the so‐called ‘climatic variability hypothesis’. However, evolutionary trade‐offs between thermal breadth and its plasticity are also seen in some taxa, and the evolution of thermal physiology remains poorly understood. Subterranean environments are excellent systems for exploring these issues, being characterized by stable climatic conditions, with environmental variability increasing predictably from deep to shallow habitats. Acclimation capacity will be fundamental in determining the sensitivity of subterranean species to climate change, since they have poor dispersal capacity and limited possibility to exploit thermally different microhabitats in the uniform cave environment. We assessed critical thermal maximum (CT_max) and short‐term heat acclimation capacity in three related beetles (Leiodidae: Leptodirini) with differing degrees of specialization to the subterranean environment (deep, shallow and facultatively subterranean, respectively) and therefore exposed to contrasting thermal variability in nature. Only the facultative subterranean species showed any acclimatory capacity, also having the highest CT_max across the taxa studied. However, this species might experience the highest thermal stress in its habitat under climate change. The studied subterranean specialists will be poorly able to cope physiologically with temperature increase, but in contrast exposed to lower magnitude and rate of warming. Our results fit the climatic variability hypothesis, suggesting that adaptation to cave conditions has selected against the retention of acclimation mechanisms. We show that the pathways that determine vulnerability of subterranean species to climate change depend on their degree of specialization to deep subterranean environments. This information, combined with evaluation of exposure to climatic changes at their present locations, is fundamental in identifying species or populations at greatest risk.

A better knowledge of the physiological basis of salinity tolerance is essential to understanding the ecology and evolutionary history of organisms that have colonized inland saline waters. Coleoptera are amongst the most diverse macroinvertebrates in inland waters, including saline habitats; however, the osmoregulatory strategies they employ to deal with osmotic stress remain unexplored. Survival and haemolymph osmotic concentration at different salinities were examined in adults of eight aquatic beetle species which inhabit different parts of the fresh—hypersaline gradient. Studied species belong to two unrelated genera which have invaded saline waters independently from freshwater ancestors; Nebrioporus (Dytiscidae) and Enochrus (Hydrophilidae). Their osmoregulatory strategy (osmoconformity or osmoregulation) was identified and osmotic capacity (the osmotic gradient between the animal’s haemolymph and the external medium) was compared between species pairs co-habiting similar salinities in nature. We show that osmoregulatory capacity, rather than osmoconformity, has evolved independently in these different lineages. All species hyperegulated their haemolymph osmotic concentration in diluted waters; those living in fresh or low-salinity waters were unable to hyporegulate and survive in hyperosmotic media (> 340 mosmol kg^-1). In contrast, the species which inhabit the hypo-hypersaline habitats were effective hyporegulators, maintaining their haemolymph osmolality within narrow limits (ca. 300 mosmol kg^-1) across a wide range of external concentrations. The hypersaline species N. ceresyi and E. jesusarribasi tolerated conductivities up to 140 and 180 mS cm^-1, respectively, and maintained osmotic gradients over 3500 mosmol kg^-1, comparable to those of the most effective insect osmoregulators known to date. Syntopic species of both genera showed similar osmotic capacities and in general, osmotic responses correlated well with upper salinity levels occupied by individual species in nature. Therefore, osmoregulatory capacity may mediate habitat segregation amongst congeners across the salinity gradient.

Exposing organims to a particular stressor may enhance tolerance to a subsequent stress, when protective mechanisms against both stressors are shared. Such cross-tolerance is a common adaptive response in dynamic multivariate environments and often indicates potential co-evolution of stress traits. Many aquatic insects in inland saline waters from Mediterranean-climate regions are sequentially challenged with salinity and desiccation stress. Thus, cross-tolerance to these physiologically similar stressors could have been positively selected in insects of these regions. We used adults of the saline water beetles Enochrus jesusarribasi (Hydrophilidae) and Nebrioporus baeticus (Dytiscidae) to test cross-tolerance responses to desiccation and salinity. In independent laboratory experiments, we evaluated the effects of i) salinity stress on the subsequent resistance to desiccation and ii) desiccation stress (rapid and slow dehydration) on the subsequent tolerance to salinity. Survival, water loss and haemolymph osmolality were measured. Exposure to stressful salinity improved water control under subsequent desiccation stress in both species, with a clear cross-tolerance (enhanced performance) in N. baeticus. In contrast, general negative effects on performance were found under the inverse stress sequence. The rapid and slow dehydration produced different water loss and haemolymph osmolality dynamics that were reflected in different survival patterns. Our finding of cross-tolerance to salinity and desiccation in ecologically similar species from distant lineages, together with parallel responses between salinity and thermal stress previously found in several aquatic taxa, highlights the central role of adaption to salinity and co-occurring stressors in arid inland waters, having important implications for the species' persistence under climate change.

Cave‐dwelling ectotherms, which have evolved for millions of years under stable thermal conditions, could be expected to have adjusted their physiological limits to the narrow range of temperatures they experience and to be highly vulnerable to global warming. However, most of the few existing studies on thermal tolerance in subterranean invertebrates highlight that despite the fact that they show lower heat tolerance than most surface‐dwelling species, their upper thermal limits are generally not adjusted to ambient temperature. The question remains to what extent this pattern is common across subterranean invertebrates. We studied basal heat tolerance and its plasticity in four species of distant arthropod groups (Coleoptera, Diplopoda, and Collembola) with different evolutionary histories but under similar selection pressures, as they have been exposed to the same constant environmental conditions for a long time. Adults were exposed at different temperatures for 1 week to determine upper lethal temperatures. Then, individuals from previous sublethal treatments were transferred to a higher temperature to determine acclimation capacity. Upper lethal temperatures of three of the studied species were similar to those reported for other subterranean species (between 20 and 25°C) and widely exceeded the cave temperature (13–14°C). The diplopod species showed the highest long‐term heat tolerance detected so far for a troglobiont (i.e., obligate subterranean) species (median lethal temperature after 7 days exposure: 28°C) and a positive acclimation response. Our results agree with previous studies showing that heat tolerance in subterranean species is not determined by environmental conditions. Thus, subterranean species, even those living under similar climatic conditions, might be differently affected by global warming.