Flight Dispersal in Supratidal Rockpool Beetles.

Flight dispersal is ecologically relevant for the survival of supratidal rockpool insects. Dispersal has important consequences for colonisation, gene flow, and evolutionary divergence. Here, we compared the flight dispersal capacity of two congeneric beetle species (Ochthebius quadricollis and Ochthebius lejolisii) that exclusively inhabit these temporary, fragmented, and extreme habitats. We estimated flight capacity and inferred dispersal in both species using different approaches: experimental flying assays, examination of wing morphology, and comparison of microsatellite markers between species. Our findings revealed that both species exhibited similar flight behaviour, with 60 to 80% of the individuals flying under water heating conditions. Notably, females of both species had larger body sizes and wing areas, along with lower wing loading, than males in O. quadricollis. These morphological traits are related to higher dispersal capacity and more energetically efficient flight, which could indicate a female-biassed dispersal pattern. The wing shapes of both species are characterised by relatively larger and narrower wings in relation to other species of the genus, suggesting high flight capacity at short distances. Molecular data revealed in both species low genetic divergences between neighbouring populations, non-significant differences between species, and no isolation by distance effect at the study scale (<100 km). These results point to passive dispersal assisted by wind.

Quantifying the performance enhancement facilitated by fractional-order implementation of classical control strategies for nanopositioning.

For most nanopositioning systems, maximizing positioning bandwidth to accurately track periodic and aperiodic reference signals is the primary performance goal. Closed-loop control schemes are employed to overcome the inherent performance limitations such as mechanical resonance, hysteresis and creep. Most reported control schemes are integer-order and combine both damping and tracking actions. In this work, fractional-order controllers from the positive position feedback family namely: the Fractional-Order Integral Resonant Control (FOIRC), the Fractional-Order Positive Position Feedback (FOPPF) controller, the Fractional-Order Positive Velocity and Position Feedback (FOPVPF) controller and the Fractional-Order Positive, Acceleration, Velocity and Position Feedback (FOPAVPF) controller are designed and analysed. Compared with their classical integer-order implementation, the fractional-order damping and tracking controllers furnish additional design (tuning) parameters, facilitating superior closed-loop bandwidth and tracking accuracy. Detailed simulated experiments are performed on recorded frequency-response data to validate the efficacy, stability and robustness of the proposed control schemes. The results show that the fractional-order versions deliver the best overall performance.

Novel Microsatellite Loci, Cross-Species Validation of Multiplex Assays, and By-Catch Mitochondrial Genomes on Ochthebius Beetles from Supratidal Rockpools.

Here we focus on designing, for the first time, microsatellite markers for evolutionary and ecological research on aquatic beetles from the genus Ochthebius (Coleoptera, Hydraenidae). Some of these non-model species, with high cryptic diversity, exclusively inhabit supratidal rockpools, extreme and highly dynamic habitats with important anthropogenic threats. We analysed 15 individuals of four species (O. lejolisii, O. subinteger, O. celatus, and O. quadricollis) across 10 localities from the Mediterranean coasts of Spain and Malta. Using next-generation sequencing technology, two libraries were constructed to interpret the species of the two subgenera present consistently (Ochthebius s. str., O. quadricollis; and Cobalius, the rest of the species). Finally, 20 markers (10 for each subgenus) were obtained and successfully tested by cross-validation in the four species under study. As a by-catch, we could retrieve the complete mitochondrial genomes of O. lejolisii, O. quadricollis, and O. subinteger. Interestingly, the mitochondrial genome of O. quadricollis exhibited high genetic variability compared to already published data. The novel SSR panels and mitochondrial genomes for Ochthebius will be valuable in future research on species identification, diversity, genetic structure, and population connectivity in highly dynamic and threatened habitats such as supratidal coastal rockpools.

Exercise-based intervention as a nonsurgical treatment for patients with carpal instability: A case series.

BACKGROUND: Although the important roles of proprioception and neuromuscular control in carpal instabilities under laboratory conditions have been recognized, only a few studies have translated this knowledge into a routine clinical practice. PURPOSE: This study aimed to evaluate the results of a personalized rehabilitation in patients with carpal instability on functionality and pain intensity. STUDY DESIGN: This was a case series study. METHODS: This case series included 39 adults (mean age: 38.2 ± 14.0 years; 16/23 females/males) diagnosed with carpal instability (radial or ulnar) with indication for orthopedic treatment. The disabilities of the arm, shoulder, and hand questionnaire was used to assess upper limb functionality. Pain perception was assessed using a visual analog scale. Exercise-based physiotherapy interventions were performed according to the clinical needs of the patients for at least 6 weeks (2-3 sessions per week). For the treatment of radial instability (n = 13), strengthening exercises of the abductor pollicis longus, extensor carpi radialis longus, flexor carpi radialis, and pronator quadratus muscles were prescribed. For the treatment of ulnar instability (n = 24), extensor carpi ulnaris and pronator quadratus were trained. All patients underwent proprioceptive training in open kinetic chain and closed kinetic chain, as well as strengthening of the unaffected hand. Changes before and after treatment were compared using the nonparametric Wilcoxon signed rank test. RESULTS: A significant improvement with a large effect size in disabilities of the arm, shoulder, and hand (P < .001; d = 2.9) and visual analog scale (P < .001; d = 3.2) scores were obtained after treatment. Moreover, the changes were greater than the minimal clinically important difference of 10.8 and 1.4, respectively. Similar results were found when patients with radial instability and ulnar instability were analyzed separately. CONCLUSIONS: Personalized training with specific proprioception and strengthening exercises produces improvements in functionality and pain perception in our cohort of people with carpal instability. These results highlight the importance of multicomponent exercise in the treatment of wrist instability. Future randomized clinical trials should further investigate the effectiveness of this protocol.

Genomic data support multiple introductions and explosive demographic expansions in a highly invasive aquatic insect.

The study of the genetic makeup and demographic fate of alien species is essential to understand their capacity to recover from founder effects, adapt to new environmental conditions and, ultimately, become invasive and potentially damaging. Here, we employ genomic data to gain insights into key demographic processes that might help to explain the extraordinarily successful invasion of the Western Mediterranean region by the North American boatman Trichocorixa verticalis (Hemiptera: Corixidae). Our analyses revealed the genetic distinctiveness of populations from the main areas comprising the invasive range and coalescent-based simulations supported that they originated from independent introductions events probably involving different source populations. Testing of alternative demographic models indicated that all populations experienced a strong bottleneck followed by a recent and instantaneous demographic expansion that restored a large portion (>30%) of their ancestral effective population sizes shortly after introductions took place (<60 years ago). Considerable genetic admixture of some populations suggest that hypothetical barriers to dispersal (i.e., land and sea water) are permeable to gene flow and/or that they originated from introductions involving multiple lineages. This study demonstrates the repeated arrival of propagules with different origins and short time lags between arrival and establishment, emphasizing the extraordinary capacity of the species to recover from founder effects and genetically admix in invaded areas. This can explain the demonstrated capacity of this aquatic insect to spread and outcompete native species once it colonizes new suitable regions. Future genomic analyses of native range populations could help to infer the genetic makeup of introduced populations and track invasion routes.

Cuticle Hydrocarbons Show Plastic Variation under Desiccation in Saline Aquatic Beetles.

In the context of aridification in Mediterranean regions, desiccation resistance and physiological plasticity will be key traits for the persistence of aquatic insects exposed to increasing desiccation stress. Control of cuticular transpiration through changes in the quantity and composition of epicuticular hydrocarbons (CHCs) is one of the main mechanisms of desiccation resistance in insects, but it remains largely unexplored in aquatic ones. We studied acclimation responses to desiccation in adults of two endemic water beetles from distant lineages living in Mediterranean intermittent saline streams: Enochrus jesusarribasi (Hydrophilidae) and Nebrioporus baeticus (Dytiscidae). Cuticular water loss and CHC composition were measured in specimens exposed to a prior non-lethal desiccation stress, allowed to recover and exposed to a subsequent desiccation treatment. E. jesusarribasi showed a beneficial acclimation response to desiccation: pre-desiccated individuals reduced cuticular water loss rate in a subsequent exposure by increasing the relative abundance of cuticular methyl-branched compounds, longer chain alkanes and branched alkanes. In contrast, N. baeticus lacked acclimation capacity for controlling water loss and therefore may have a lower physiological capacity to cope with increasing aridity. These results are relevant to understanding biochemical adaptations to drought stress in inland waters in an evolutionary and ecological context.

DISPERSE, a trait database to assess the dispersal potential of European aquatic macroinvertebrates.

Dispersal is an essential process in population and community dynamics, but is difficult to measure in the field. In freshwater ecosystems, information on biological traits related to organisms' morphology, life history and behaviour provides useful dispersal proxies, but information remains scattered or unpublished for many taxa. We compiled information on multiple dispersal-related biological traits of European aquatic macroinvertebrates in a unique resource, the DISPERSE database. DISPERSE includes nine dispersal-related traits subdivided into 39 trait categories for 480 taxa, including Annelida, Mollusca, Platyhelminthes, and Arthropoda such as Crustacea and Insecta, generally at the genus level. Information within DISPERSE can be used to address fundamental research questions in metapopulation ecology, metacommunity ecology, macroecology and evolutionary ecology. Information on dispersal proxies can be applied to improve predictions of ecological responses to global change, and to inform improvements to biomonitoring, conservation and management strategies. The diverse sources used in DISPERSE complement existing trait databases by providing new information on dispersal traits, most of which would not otherwise be accessible to the scientific community.

Role of cuticle hydrocarbons composition in the salinity tolerance of aquatic beetles.

Salinity tolerance has enabled the colonization of inland saline waters and promoted species diversification in some lineages of aquatic insects. However, the mechanisms behind this tolerance, particularly the role of cuticle hydrocarbons (CHCs), are not well-known. We characterized the CHC profile of eight species of two water beetle genera (Nebrioporus, Adephaga: Dytiscidae and Enochrus, Polyphaga: Hydrophilidae), which span the fresh-hypersaline gradient, to: i) determine the interspecific variation of CHC composition in relation to species' salinity tolerance; ii) explore plastic adjustments in CHC profiles in response to salinity changes at the intraspecific level in saline-tolerant species. CHC profiles were highly species-specific, more complex and diverse in composition, and characterized by longer-chain-length compounds in the species with higher salinity tolerance within each genus. Higher salinity tolerance in the Enochrus species was also associated with an increase in the relative abundance of branched alkanes, and with a lower proportion of n-alkanes and unsaturated compounds. These CHC characteristics are related with improved waterproofing capacity and suggest that reducing cuticle permeability was one of the key mechanisms to adapt to saline waters. Similar CHC composition patterns were found at the intraspecific level between populations from lower and higher salinity sites within saline-tolerant species of each genus. These saline species also displayed an extraordinary ability to adjust CHC profiles to changing salinity conditions in the laboratory in a relatively short time, which reflects great plasticity and a high potential to deal with daily and seasonal environmental fluctuations in the highly dynamic saline habitats.

Comparación de la fuerza de unión y la integridad marginal con compuestos de resina de relleno masivo y compuestos indirectos / Comparing bond strength and marginal integrity with direct bulk-fill resin composites and indirect composites

ABSTRACT Introduction: The clinical longevity of tooth restoration -whether directly or indirectly using composites- greatly depends on the quality and stability of the marginal adaptation. Even today, dental restoration failure is a major complication in everyday dental practice. Objective: To evaluate the effect of restoration techniques on the microtensile bond strength and marginal integrity of class II cavities. Methods: An experimental in vitro investigation was made. Preparations (5 × 4 × 2 mm) below the cement-enamel junction were performed in 45 human maxillary premolars (n= 15) that were the sample of the study selected to random. The G1 group incrementally received Spectrum TPH3 Dentsply De trey in three horizontal incremental layers. The G2 group received a bulk restoration technique (one 4-mm increment of Surefill SDR flow plus one 1-mm horizontal capping layer of Spectrum TPH3 Dentsply De trey using a metal matrix band. For the G3 group, impressions were made from each cavity preparation, and Spectrum was used to complete an indirect composite restoration. After storage (24 h/37 °C), the proximal surfaces of each tooth were polished with Sof-Lex disks. For microtensile bond strength testing, all premolars were sectioned into resin-dentine beams (0.8 mm2) and were tested under tension (0.5 mm/min). Results: Microtensile bond strength testing and marginal integrity values were not statistically significantly affected by the type of restoration technique used (p> 0.05). Conclusions: The Surefill SDR flow that used a capping layer made of conventional composite can be an alternative to reduce procedure durations as well as additional steps in the restorative technique(AU)

RESUMEN Introducción: La longevidad clínica de una restauración dental -utilizando compuestos bien directa o indirectamente- depende en gran medida de la calidad y la estabilidad de la adaptación marginal. Incluso hoy en día las restauraciones dentales fallidas constituyen una importante complicación en la práctica dental cotidiana. Objetivo: Evaluar el efecto de las técnicas de restauración en la fuerza de unión microtensil y la integridad marginal de las cavidades clase II. Métodos: Se llevó a cabo una investigación experimental in vitro. Se realizaron preparaciones (5 × 4 × 2 mm) por debajo de la unión cemento-esmalte en 45 premolares maxilares humanos (n= 15), los que constituyeron la muestra aleatoria del estudio. El Grupo G1 recibió incrementalmente Spectrum TPH3 Dentsply (De Trey) en tres capas horizontales incrementales. El Grupo G2 recibió una técnica de restauración masiva (un incremento de 4-mm de flujo de SureFil SDR más una capa de tapado horizontal de 1-mm de Spectrum TPH3 Dentsply (De Trey) utilizando una banda matriz metálica. En el Grupo G3 se realizaron impresiones de la preparación de cada cavidad, y se usó Spectrum para completar una restauración indirecta con compuesto. Después del almacenamiento (24 h / 37 °C), se pulieron las superficies proximales de cada diente con discos Sof-Lex. Para evaluar la fuerza de unión microtensil, todos los premolares fueron seccionados en haces de resina-dentina (0,8 mm2) y fueron examinados bajo tensión (0,5 mm/min). Resultados: Las pruebas de fuerza de unión microtensil y los valores de integridad marginal no fueron afectados significativamente desde el punto de vista estadístico por el tipo de técnica de restauración utilizado (p> 0,05). Conclusiones: El flujo de SureFil SDR que emplea una capa de tapado hecha de compuesto convencional puede ser una alternativa para reducir la duración del procedimiento, así como los pasos adicionales de la técnica de restauración(AU)

Two-degrees-of-freedom PI2D controller for precise nanopositioning in the presence of hardware-induced constant time delay.

The fast and accurate tracking of periodic and arbitrary reference trajectories is the principal goal in many nanopositioning applications. Flexure-based piezoelectric stack driven nanopositioners are widely employed in applications where accurate mechanical displacements at these nanometer scales are required. The performance of these nanopositioners is limited by the presence of lightly damped resonances in their dynamic response and actuator nonlinearities. Closed-loop control techniques incorporating both damping and tracking are typically used to address these limitations. However, most tracking schemes employed use a first-order integrator where a triangular trajectory commonly used in nanopositioning applications necessitates a double integral for zero-error tracking. The phase margin of the damped system combined with the hardware-induced delay deem the implementation of a double-integrator unstable. To overcome this limitation, this paper presents the design, analysis and application of a new control scheme based on the structure of the traditional Two-Degrees-of-Freedom PID controller (2DOF-PID). The proposed controller replaces the integral action of the traditional 2DOF-PID with a double integral action (2DOF-PI2D). Despite its simplicity, the proposed controller delivers superior tracking performance compared to traditional combined damping and tracking control schemes based on well-reported designs such as positive position feedback (PPF), Integral resonant control (IRC), and Positive Velocity and Position Feedback (PVPF). The stability of the control system is analyzed in the presence of a time delay in the system. Experimental results validating the efficacy of the proposed chattering-free control of a piezo-driven nanopositioning system are included.

Evaluating anthropogenic impacts on naturally stressed ecosystems: Revisiting river classifications and biomonitoring metrics along salinity gradients.

Naturally stressed ecosystems hold a unique fraction of biodiversity. However, they have been largely ignored in biomonitoring and conservation programmes, such as the EU Water Framework Directive, while global change pressures are threatening their singular values. Here we present a framework to classify and evaluate the ecological quality of naturally stressed rivers along a water salinity gradient. We gathered datasets, including aquatic macroinvertebrate assemblages and environmental information, for 243 river locations across the western Mediterranean to: a) gauge the role of natural stressors (salinity) in driving aquatic community richness and composition; b) make river classifications by encompassing the wide range of environmental and biological variation exhibited by Mediterranean rivers; c) provide effective biomonitoring metrics of ecological quality for saline rivers. Our results showed that water salinity played a pivotal role in explaining the community richness and compositional changes in rivers, even when considering other key and commonly used descriptors, such as elevation, climate or lithology. Both environmental and biologically-based classifications included seven river types: three types of freshwater perennial rivers, one freshwater intermittent river type and three new saline river types. These new saline types were not included in previous classifications. Their validation by independent datasets showed that the saline and freshwater river types represented differentiable macroinvertebrate assemblages at family and species levels. Biomonitoring metrics based on the abundance of indicator taxa of each saline river type provided a much better assessment of the ecological quality of saline rivers than other widely used biological metrics and indices. Here we demonstrate that considering natural stressors, such as water salinity, is essential to design effective and accurate biomonitoring programmes for rivers and to preserve their unique biodiversity.

Do all roads lead to Rome? Exploring community trajectories in response to anthropogenic salinization and dilution of rivers.

Abiotic stress shapes how communities assemble and support ecological functions. However, it remains unclear whether artificially increasing or decreasing stress levels would lead to communities assembling predictably along a single axis of variation or along multiple context-dependent trajectories of change. In response to stress intensity alterations, we hypothesize that a single trajectory of change occurs when trait-based assembly prevails, while multiple trajectories of change arise when dispersal-related processes modify colonization and trait-filtering dynamics. Here, we tested these hypotheses using aquatic macroinvertebrates from rivers exposed to gradients of natural salinity and artificially diluted or salinized ion contents. Our results showed that trait-filtering was important in driving community assembly in natural and diluted rivers, while dispersal-related processes seemed to play a relevant role in response to salinization. Salinized rivers showed novel communities with different trait composition, while natural and diluted communities exhibited similar taxonomic and trait compositional patterns along the conductivity gradient. Our findings suggest that the artificial modification of chemical stressors can result in different biological communities, depending on the direction of the change (salinization or dilution), with trait-filtering, and organism dispersal and colonization dynamics having differential roles in community assembly. The approach presented here provides both empirical and conceptual insights that can help in anticipating the ecological effects of global change, especially for those stressors with both natural and anthropogenic origins.This article is part of the theme issue 'Salt in freshwaters: causes, ecological consequences and future prospects'.

Insect communities in saline waters consist of realized but not fundamental niche specialists.

Considering how organisms adapt to stress is essential if we are to anticipate biological responses to global change in ecosystems. Communities in stressful environments can potentially be assembled by specialists (i.e. species that only occur in a limited range of environmental conditions) and/or generalist species with wider environmental tolerances. We review the existing literature on the salinity tolerance of aquatic insects previously identified as saline specialists because they were exclusively found in saline habitats, and explore if these saline realized niche specialists are also specialists in their fundamental niches or on the contrary are fundamental niche generalist species confined to the highest salinities they can tolerate. The results suggest that species inhabiting saline waters are generalists in their fundamental niches, with a predominant pattern of high survival in freshwater-low salinity conditions, where their fitness tends to be similar or even higher than in saline waters. Additionally, their performance in freshwater tends to be similar to related strictly freshwater species, so no apparent trade-off of generalization is shown. These results are discussed in the framework of the ecological and evolutionary processes driving community assembly across the osmotic stress gradient, and their potential implications for predicting impacts from saline dilution and freshwater salinization.This article is part of the theme issue 'Salt in freshwaters: causes, ecological consequences and future prospects'.

Effects of salinity changes on aquatic organisms in a multiple stressor context.

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'.

New data on the distribution of aquatic bugs (Hemiptera) from eastern Morocco with notes on chorology.

New faunistic and distributional data on aquatic Hemiptera from the Oriental Region of Morocco and the Moulouya River basin, covering an area of 119,268 km2, are presented. An annotated list of 45 species, 21 genera and 11 families is provided. Nine species are new records for the entire studied area, and two for the Moulouya River basin. We rediscovered Velia noualhieri iberica and Notonecta viridis after eight decades. We also provide novel records for Aphelocheirus pemae and Hebrus atlas, which were recently described from Morocco, and for the invasive corixid Trichocorixa verticalis verticalis. The dominance of species with broad distributions points to the high dispersal dynamics of the Hemiptera community in the study area, similar to that of other areas where a semiarid climate prevails.

Fractional order implementation of Integral Resonant Control - A nanopositioning application.

By exploiting the co-located sensor-actuator arrangement in typical flexure-based piezoelectric stack actuated nanopositioners, the polezero interlacing exhibited by their axial frequency response can be transformed to a zero-pole interlacing by adding a constant feed-through term. The Integral Resonant Control (IRC) utilizes this unique property to add substantial damping to the dominant resonant mode by the use of a simple integrator implemented in closed loop. IRC used in conjunction with an integral tracking scheme, effectively reduces positioning errors introduced by modelling inaccuracies or parameter uncertainties. Over the past few years, successful application of the IRC control technique to nanopositioning systems has demonstrated performance robustness, easy tunability and versatility. The main drawback has been the relatively small positioning bandwidth achievable. This paper proposes a fractional order implementation of the classical integral tracking scheme employed in tandem with the IRC scheme to deliver damping and tracking. The fractional order integrator introduces an additional design parameter which allows desired pole-placement, resulting in superior closed loop bandwidth. Simulations and experimental results are presented to validate the theory. A 250% improvement in the achievable positioning bandwidth is observed with proposed fractional order scheme.

Wiener-Hopf optimal control of a hydraulic canal prototype with fractional order dynamics.

This article addresses the control of a laboratory hydraulic canal prototype that has fractional order dynamics and a time delay. Controlling this prototype is relevant since its dynamics closely resembles the dynamics of real main irrigation canals. Moreover, the dynamics of hydraulic canals vary largely when the operation regime changes since they are strongly nonlinear systems. All this makes difficult to design adequate controllers. The controller proposed in this article looks for a good time response to step commands. The design criterium for this controller is minimizing the integral performance index ISE. Then a new methodology to control fractional order processes with a time delay, based on the Wiener-Hopf control and the Padé approximation of the time delay, is developed. Moreover, in order to improve the robustness of the control system, a gain scheduling fractional order controller is proposed. Experiments show the adequate performance of the proposed controller.

The chicken or the egg? Adaptation to desiccation and salinity tolerance in a lineage of water beetles.

Transitions from fresh to saline habitats are restricted to a handful of insect lineages, as the colonization of saline waters requires specialized mechanisms to deal with osmotic stress. Previous studies have suggested that tolerance to salinity and desiccation could be mechanistically and evolutionarily linked, but the temporal sequence of these adaptations is not well established for individual lineages. We combined molecular, physiological and ecological data to explore the evolution of desiccation resistance, hyporegulation ability (i.e., the ability to osmoregulate in hyperosmotic media) and habitat transitions in the water beetle genus Enochrus subgenus Lumetus (Hydrophilidae). We tested whether enhanced desiccation resistance evolved before increases in hyporegulation ability or vice versa, or whether the two mechanisms evolved in parallel. The most recent ancestor of Lumetus was inferred to have high desiccation resistance and moderate hyporegulation ability. There were repeated shifts between habitats with differing levels of salinity in the radiation of the group, those to the most saline habitats generally occurring more rapidly than those to less saline ones. Significant and accelerated changes in hyporegulation ability evolved in parallel with smaller and more progressive increases in desiccation resistance across the phylogeny, associated with the colonization of meso- and hypersaline waters during global aridification events. All species with high hyporegulation ability were also desiccation-resistant, but not vice versa. Overall, results are consistent with the hypothesis that desiccation resistance mechanisms evolved first and provided the physiological basis for the development of hyporegulation ability, allowing these insects to colonize and diversify across meso- and hypersaline habitats.

Cuticle hydrocarbons in saline aquatic beetles.

Hydrocarbons are the principal component of insect cuticle and play an important role in maintaining water balance. Cuticular impermeability could be an adaptative response to salinity and desiccation in aquatic insects; however, cuticular hydrocarbons have been poorly explored in this group and there are no previous data on saline species. We characterized cuticular hydrocarbons of adults and larvae of two saline aquatic beetles, namely Nebrioporus baeticus (Dytiscidae) and Enochrus jesusarribasi (Hydrophilidae), using a gas chromatograph coupled to a mass spectrometer. The CHC profile of adults of both species, characterized by a high abundance of branched alkanes and low of unsaturated alkenes, seems to be more similar to that of some terrestrial beetles (e.g., desert Tenebrionidae) compared with other aquatic Coleoptera (freshwater Dytiscidae). Adults of E. jesusarribasi had longer chain compounds than N. baeticus, in agreement with their higher resistance to salinity and desiccation. The more permeable cuticle of larvae was characterized by a lower diversity in compounds, shorter carbon chain length and a higher proportion of unsaturated hydrocarbons compared with that of the adults. These results suggest that osmotic stress on aquatic insects could exert a selection pressure on CHC profile similar to aridity in terrestrial species.

Aquatic insects in a multistress environment: cross-tolerance to salinity and desiccation.

Exposing organisms to a particular stressor may enhance tolerance to a subsequent stress, when protective mechanisms against the two 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.