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Predators often search for prey while moving through the environment, but there are important exceptions, including the way sedentary predators sometimes rely on signals for drawing prey to within striking distance 1 , 2 . Some spiders, for instance, leave the remnants of previously-captured prey in their webs where they function as static lures that effectively attract a diverse array of additional prey 3 4 5 6 . However, important questions remain concerning how specific the targeted prey may be and how dynamic, instead of static, signalling might be. With these questions as our rationale, we initiated research on Araneus ventricosus (L. Koch, 1878), an orb-weaving spider, as the predator and the firefly Abscondita terminalis males as the prey (Figure 1A-C). Using two lanterns situated on their abdomen (Figure 1D,F), A. terminalis males make female-attracting multi-pulse flash trains (Figure 1J), whereas sedentary females attract males by making single-pulse signals (Figure 1C,K) with a single lantern (Figure 1E,G). Drawing from extensive field observations, we propose that A. ventricosus practices deceptive interspecific communication by first ensnaring firefly males in its web and then predisposing the entrapped male fireflies to broadcast bioluminescent signals that deviate from female-attracting signals typically made by A. terminalis males and instead mimic the male-attracting signals typically made by females. The outcome is that the entrapped male fireflies broadcast false signals that lure more male fireflies into the web., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Ruderal plants are an important component of plant communities that develop on the range of anthropogenically degraded lands. Yet they were highly neglected and not recognised as desirable for restoration purposes. The aim of this study was to analyse the potential for using ruderal species in restoration processes and to identify preliminary criteria for species selection that could be included in ecological restoration of degraded man-made habitats under future conditions of increased human disturbance and climate changes. The desirable characteristics of the species depend primarily on the type of habitat to be restored, with plant height, specific leaf area, rooting depth and seed characteristics being the most important traits. The recognised ecosystem services of the species analysed show that the provisioning and regulating services are well represented, particularly erosion control, pollination, phytoremediation and other soil quality improvements. Most of the dominant and diagnostic ruderal species from the man-made habitats of the north-western Balkan Peninsula are sensitive to climate change and their potential distribution range is expected to decrease at the European scale. Higher certain ecological indicator values, as well as values for disturbance severity, frequency and soil disturbance indices were found for species that are expected to increase their range. Ruderal species are becoming increasingly important for restoration purposes, as the focus shifts to the significance of early successional species. The inclusion of ruderal species in the restoration of degraded sites should be based on criteria such as: non-invasiveness, plant traits favourable for colonisation (height, SLA, seed traits, rooting depth), values of ecological and disturbance indices, provision of ecosystem services, and change of distribution range under changing climate conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Particulate matter concentrations (PM 10 , PM 2.5 , PM 1 ) and microclimatic parameters (air temperature, CO 2 ) were monitored in the Škocjan Caves (Slovenia). The effects of tourist visits on the PM concentrations and the cave's microclimate are immediate and direct, but these values normalise relatively quickly. The results showed seasonal, diurnal, and spatial differences in all parameters studied. Due to the higher number of visitors, their influence on the cave's microclimate and PM 10 and PM 2.5 concentrations is greater in summer than in winter. The measured PM 1 levels depend on the ventilation in the cave, as air transport plays an important role in their introduction into the cave. PM consists of minerals of natural origin resulting from the re-suspension of cave sediments due to strong air currents generated by the opening of the doors to tourists and their walks. The second most common influence is the anthropogenic phases originating from maintenance work in the cave, electronic devices, cave lighting and emissions from outside the cave (aerosols from the polluted Reka River, industry, traffic, gypsum waste disposal). In order to upgrade the sustainable use of the UNESCO-listed Škocjan Caves for tourism, we propose regular monitoring of PM and a detailed characterization of individual PMs and their sources, in addition to monitoring of the cave's microclimate and biology., (© 2024. The Author(s).)

Sexual dimorphism can evolve in response to sex-specific selection pressures that vary across habitats. We studied sexual differences in subterranean amphipods Niphargus living in shallow subterranean habitats (close to the surface), cave streams (intermediate), and cave lakes (deepest and most isolated). These three habitats differ because at greater depths there is lower food availability, reduced predation, and weaker seasonality. Additionally, species near the surface have a near-even adult sex ratio (ASR), whereas species from cave lakes have a female-biased ASR. We hypothesized (a) a decrease in sexual dimorphism from shallow subterranean habitats to cave lake species because of weaker sexual selection derived from changes in the ASR and (b) an increase in female body size in cave lakes because of stronger fecundity selection on account of oligotrophy, reduced predation, and weaker seasonality. We measured body size and two sexually dimorphic abdominal appendages for all 31 species and several behaviours related to male competition (activity, risk-taking, exploration) for 12 species. Species with an equal ASR that live close to the surface exhibited sexual dimorphism in all three morphological traits, but not in behaviour. The body size of females increased from the surface to cave lakes, but no such trend was observed in males. In cave lake species, males and females differed neither morphologically nor behaviourally. Our results are consistent with the possibility that sexual and fecundity selection covary across the three habitats, which indirectly and directly, respectively, shape the degree of sexual dimorphism in Niphargus species., (© The Author(s) 2024. Published by Oxford University Press on behalf of the European Society of Evolutionary Biology.)

Mating plugs in animals are ubiquitous and are commonly interpreted to be products of mating strategies. In spiders, however, mating plugs may also take on functions beyond female remating prevention. Due to the vagaries of female genital (spermathecal) anatomy, most spiders face the problem of having to secure additional, non-anatomical, protection for transferred sperm. Here, we test the hypothesis that mating plugs, rather than (or in addition to) being adaptations for mating strategies, may serve as sperm protection mechanism. Based on a comparative study on 411 epigyna sampled from 36 families, 187 genera, 330 species of entelegyne spiders, our results confirm the necessity of a sperm protection mechanism. We divided the entelegyne spermathecae into four types: SEG, SED, SCG and SCD. We also studied detailed morphology of epigynal tracts in the spider Diphya wulingensis having the SEG type spermathecae, using 3D-reconstruction based on semi thin histological series section. In this species, we hypothesize that two distinct types of mating plug, the sperm plug and the secretion plug, serve different functions. Morphological details support this: sperm plugs are formed on a modified spermathecal wall by the spilled sperm, and function as a temporary protection mechanism to prevent sperm from leaking and desiccating, while secretion plugs function in postcopulation both as a permanent protection mechanism, and to prevent additional mating. Furthermore, with the modified spermathecal wall of S2 stalk, the problem of shunt of sperm input and output, and the possibility of female multiple mating have been resolved. Variation in spermathecal morphology also suggests that the problem of sperm protection might be resolved in different ways in spiders. Considering mating plugs of varying shapes and origins in the vast morphospace of spiders, we conclude that mating plugs might serve different purposes that relate both to mating strategies, as well as to sperm protection., Competing Interests: the authors have declared that no competing interests exist., (Copyright: © 2024 Jiang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Nonhuman animal protagonists of folklore texts in the European space have tended to be perceived primarily as performing a symbolic and metaphoric function. But behind the symbols and the metaphors hide real flesh-and-blood nonhuman animals, and flesh-and-blood humans interacting with them, mostly from a position of power. The emerging discipline of zoofolkloristics considers nonhuman animals in their own right. Through critical analysis of folklore material, zoofolkloristics examines the role of animals and power relations within the interspecies entanglement with the aim of deconstructing the oppressive system and establishing multispecies justice. We begin this paper with a brief reflection on the 'historical animal' as an embodied being and a human construct. We then perform a critical re-reading of three animal-related folklore texts from the Slovenian tradition and, applying Hubert Zapf's concept of imaginative counter-discourse, consider the potential of imagination as a methodological tool in the transformative program of zoofolkloristics. Implications for animal ethics, liberation, and conservation are also discussed.

Madagascar is a global biodiversity hotspot, but its biodiversity continues to be underestimated and understudied. Of raft spiders, genus Dolomedes Latreille, 1804, literature only reports two species on Madagascar. Our single expedition to humid forests of eastern and northern Madagascar, however, yielded a series of Dolomedes exemplars representing both sexes of five morphospecies. To avoid only using morphological diagnostics, we devised and tested an integrative taxonomic model for Dolomedes based on the unified species concept. The model first determines morphospecies within a morphometrics framework, then tests their validity via species delimitation using COI. It then incorporates habitat preferences, geological barriers, and dispersal related traits to form hypotheses about gene flow limitations. Our results reveal four new Dolomedes species that we describe from both sexes as Dolomedes gregoric sp. nov. , D. bedjanic sp. nov. , D. hydatostella sp. nov. , and D. rotundus sp. nov. The range of D. kalanoro Silva & Griswold, 2013, now also known from both sexes, is expanded to eastern Madagascar. By increasing the known raft spider diversity from one valid species to five, our results merely scratch the surface of the true Dolomedes species diversity on Madagascar. Our integrative taxonomic model provides the framework for future revisions of raft spiders anywhere., Competing Interests: The authors declare that they have no competing interests., (© 2024 Yu and Kuntner.)

Fine particulate matter (PM 2.5 ), a detrimental urban air pollutant primarily emitted by traffic and biomass burning, poses disproportionately significant health risks at relatively limited exposure during commuting. Previous studies have mainly focused on fixed locations when assessing PM 2.5 exposure, while neglecting pedestrians and cyclists, who often experience higher pollution levels. In response, this research aimed to independently validate the effectiveness of bicycle-mounted low-cost sensors (LCS) adopted by citizens, evaluate temporal and spatial PM 2.5 exposure, and assess associated health risks in Ljubljana, Slovenia. The LCS quality assurance results, verified by co-location field tests by air quality monitoring stations (AQMS), showed comparable outcomes with an average percentage difference of 21.29 %, attributed to humidity-induced nucleation effects. The colder months exhibited the highest air pollution levels (μ = 32.31 μg/m 3 ) due to frequent thermal inversions and weak wind circulation, hindering vertical air mixing and the adequate dispersion of pollutants. Additionally, PM 2.5 levels in all sampling periods were lowest in the afternoon (μ = 12.09 μg/m 3 ) and highest during the night (μ = 61.00 μg/m 3 ) when the planetary boundary layer thins, leading to the trapping of pollutants near the surface, thus significantly affecting diurnal and seasonal patterns. Analysis of exposure factors revealed that cyclists were approximately three times more exposed than pedestrians. However, the toxicological risk assessment indicated a minimal potential risk of PM 2.5 exposure. The collaborative integration of data from official AQMS and LCS can enhance evidence-based policy-making processes and facilitates the realignment of effective regulatory frameworks to reduce urban air pollution., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Transition to novel environments, such as groundwater colonization by surface organisms, provides an excellent research ground to study phenotypic evolution. However, interspecific comparative studies on evolution to groundwater life are few because of the challenge in assembling large ecological and molecular resources for species-rich taxa comprised of surface and subterranean species. Here, we make available to the scientific community an operational set of working tools and resources for the Asellidae, a family of freshwater isopods containing hundreds of surface and subterranean species. First, we release the World Asellidae database (WAD) and its web application, a sustainable and FAIR solution to producing and sharing data and biological material. WAD provides access to thousands of species occurrences, specimens, DNA extracts and DNA sequences with rich metadata ensuring full scientific traceability. Second, we perform a large-scale dated phylogenetic reconstruction of Asellidae to support phylogenetic comparative analyses. Of 424 terminal branches, we identify 34 pairs of surface and subterranean species representing independent replicates of the transition from surface water to groundwater. Third, we exemplify the usefulness of WAD for documenting phenotypic shifts associated with colonization of subterranean habitats. We provide the first phylogenetically controlled evidence that body size of males decreases relative to that of females upon groundwater colonization, suggesting competition for rare receptive females selects for smaller, more agile males in groundwater. By making these tools and resources widely accessible, we open up new opportunities for exploring how phenotypic traits evolve in response to changes in selective pressures and trade-offs during groundwater colonization., (© 2023 John Wiley & Sons Ltd.)

Sexual dimorphism, or sex-specific trait expression, may evolve when selection favours different optima for the same trait between sexes, that is, under antagonistic selection. Intra-locus sexual conflict exists when the sexually dimorphic trait under antagonistic selection is based on genes shared between sexes. A common assumption is that the presence of sexual-size dimorphism (SSD) indicates that sexual conflict has been, at least partly, resolved via decoupling of the trait architecture between sexes. However, whether and how decoupling of the trait architecture between sexes has been realized often remains unknown. We tested for differences in architecture of adult body size between sexes in a species with extreme SSD, the African hermit spider (Nephilingis cruentata), where adult female body size greatly exceeds that of males. Specifically, we estimated the sex-specific importance of genetic and maternal effects on adult body size among individuals that we laboratory-reared for up to eight generations. Quantitative genetic model estimates indicated that size variation in females is to a larger extent explained by direct genetic effects than by maternal effects, but in males to a larger extent by maternal than by genetic effects. We conclude that this sex-specific body-size architecture enables body-size evolution to proceed much more independently than under a common architecture to both sexes., (© 2023 The Authors. Journal of Evolutionary Biology published by John Wiley & Sons Ltd on behalf of European Society for Evolutionary Biology.)

In our study, we set out to collect a multimodal annotated dataset for remote sensing of Maya archaeology, that is suitable for deep learning. The dataset covers the area around Chactún, one of the largest ancient Maya urban centres in the central Yucatán Peninsula. The dataset includes five types of data records: raster visualisations and canopy height model from airborne laser scanning (ALS) data, Sentinel-1 and Sentinel-2 satellite data, and manual data annotations. The manual annotations (used as binary masks) represent three different types of ancient Maya structures (class labels: buildings, platforms, and aguadas - artificial reservoirs) within the study area, their exact locations, and boundaries. The dataset is ready for use with machine learning, including convolutional neural networks (CNNs) for object recognition, object localization (detection), and semantic segmentation. We would like to provide this dataset to help more research teams develop their own computer vision models for investigations of Maya archaeology or improve existing ones., (© 2023. Springer Nature Limited.)

Water suppliers should provide safe drinking water following preventive measures. This is especially important for karst water sources, as they are among the most vulnerable. Recently, there has been a strong focus on the early warning system, which mainly involves monitoring proxy parameters, but does not consider drainage area conditions and other monitoring recommendations. Here, we present an innovative strategy for assessing contamination risk of karst water sources that covers spatio-temporal dimensions and can be integrated into management practices. It is based on event-based monitoring and risk mapping and has been tested in a well-known study area. The holistic early warning system provides accurate spatial hazard and risk assessment and operational monitoring guidelines, including locations, indicator parameters, and temporal resolution and duration. In the study area, the high contamination risk, representing 0.5 % of the area, was spatially delineated. The highest probability of source contamination occurs during recharge events when proxy parameters such as bacteria, ATP, Cl, and Ca/Mg ratio should be monitored in addition to continuous monitoring of turbidity, EC, and T. Monitoring of sinking streams should serve as a preventive measure, since water transfer from ponors to springs has been shown to take about one day, and poor quality water is present for at least another day. Therefore, intensive monitoring should be conducted at intervals of a few hours for at least a week. Although hydrologic systems vary, the proposed strategy is particularly useful in systems where water flows rapidly and where remediation is not feasible., Competing Interests: Declaration of competing interest The authors of the manuscript certify that they have no affiliations with or involvement in any organization or entity with any financial interest (such as honoraria; educational grants; participation in speakers' bureaus; membership, employment, consultancies, stock ownership, or other equity interest; and expert testimony or patent-licensing arrangements), or non-financial interest (such as personal or professional relationships, affiliations, knowledge or beliefs) in the subject matter or materials discussed in this manuscript., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

This study investigated the use of affect and physiological signals of heart rate, electrodermal activity, pupil dilation, and skin temperature to classify advertising engagement. The ground truth for the affective and behavioral aspects of ad engagement was collected from 53 young adults using the User Engagement Scale. Three gradient-boosting classifiers, LightGBM (LGBM), HistGradientBoostingClassifier (HGBC), and XGBoost (XGB), were used along with signal fusion to evaluate the performance of different signal combinations as predictors of engagement. The classifiers trained on the fusion of skin temperature, valence, and tiredness (features n = 5) performed better than those trained on all signals (features n = 30). The average AUC ROC scores for the fusion set were XGB = 0.68 (0.10), LGBM = 0.69 (0.07), and HGBC = 0.70 (0.11), compared to the lower scores for the set of all signals (XGB = 0.65 (0.11), LGBM = 0.66 (0.11), HGBC = 0.64 (0.10)). The results also show that the signal fusion set based on skin temperature outperforms the fusion sets of the other three signals. The main finding of this study is the role of specific physiological signals and how their fusion aids in more effective modeling of ad engagement while reducing the number of features.

Caves and their surrounding fracture systems in the vadose zone of karst regions host a unique atmospheric environment. Understanding the airflow patterns in caves is critical to understanding the properties of the subsurface atmosphere and the chemical interactions between air, water, and rock. The most common driver of airflow in caves is the density difference between the subsurface and the outside air, known as the chimney effect. Observations show that seasonal airflow patterns in caves also depend on the geometry of passages. In this work, I present and use a numerical model of a passage embedded and thermally coupled to a rock mass to study the relationship between the airflow pattern and passage geometry. As the outside air enters the subsurface, it approaches thermal equilibrium with the rock mass along a characteristic relaxation length. This determines the temperature and density contrast between the inside and outside air, and the resulting pressure difference, which drives the airflow. In passages with non-uniform outlines and/or cross-sections, the relaxation length may depend on the flow direction, resulting in different airflow velocities in cold and warm periods for the same absolute temperature difference between the massif and the external temperature. In a passage with a V-shaped longitudinal profile, the airflow is triggered by instability which causes the feedback between the relaxation length and airflow velocity. The airflow pattern can also be altered by snow and ice. Heat transfer in the rock and the thermal inertia of the rock also change the relaxation lengths and cause hysteresis in the curve presenting the airflow velocity vs. temperature difference., Competing Interests: The author has declared that no competing interests exist., (Copyright: © 2023 Franci Gabrovšek. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Ecological theory predicts close relationships between macroclimate and functional traits. Yet, global climatic gradients correlate only weakly with the trait composition of local plant communities, suggesting that important factors have been ignored. Here, we investigate the consistency of climate-trait relationships for plant communities in European habitats. Assuming that local factors are better accounted for in more narrowly defined habitats, we assigned > 300,000 vegetation plots to hierarchically classified habitats and modelled the effects of climate on the community-weighted means of four key functional traits using generalized additive models. We found that the predictive power of climate increased from broadly to narrowly defined habitats for specific leaf area and root length, but not for plant height and seed mass. Although macroclimate generally predicted the distribution of all traits, its effects varied, with habitat-specificity increasing toward more narrowly defined habitats. We conclude that macroclimate is an important determinant of terrestrial plant communities, but future predictions of climatic effects must consider how habitats are defined., (© 2023. The Author(s).)

It is assumed that people practiced woodland management, i.e., coppicing and pollarding, in prehistory, but details are poorly known. This study aims for a better understanding of woodland exploitation through time in the wetland basin of the Ljubljansko barje, Slovenia, from 3700-2400 BCE (Before Common Era). To do so, uncarbonized, waterlogged wood from 16 Eneolithic pile dwellings situated in two geographical clusters that cover a time span of c. 1300 years were subjected to age/diameter analysis. It is the first time that age/diameter analysis has been applied to multiple sites from the same region. The investigated posts represent a wide range of taxa, but oak ( Quercus sp.) and ash ( Fraxinus sp.) represent 75% of the total, indicating selective use of wood for this purpose. Diameter selection of ash may have taken place as well. At both site clusters, the age/diameter data do not reveal any unequivocal evidence for woodland management. Only at the youngest sites do the data possibly show some gradually changing practices. The outcomes are discussed within the framework of recent discussions about woodland management in Europe.

Archaeoastronomical studies have demonstrated that the important civic and ceremonial buildings in Mesoamerica were largely oriented to sunrises or sunsets on specific dates, but the origin and spread of orientation practices were not clear. Using aerial laser scanning (lidar) data, we analyzed orientations of a large number of ceremonial complexes in the area along the southern Gulf Coast, including many recently identified Formative sites dating to 1100 BCE to 250 CE. The distribution pattern of dates marked by solar alignments indicates their subsistence-related ritual significance. The orientations of complexes built between 1100 and 750 BCE, in particular, represent the earliest evidence of the use of the 260-day calendar, centuries earlier than its previously known use in textual records.

The current COVID-19 pandemic caught the decision makers in many countries sub-optimally prepared to respond. To better cope with similar situations in the future, it is vital to understand the major predictors of health-beneficial behavior and adherence to imposed mitigation measures and guidelines. To tailor the promotion of government-imposed measures, it is important to understand how the sociodemographic background combined with personality traits affect the perception and responsiveness of people. We investigated the perception and adherence to mitigation measures during the pandemic by examining their trends across several sociodemographic categories and personality dimensions. The strongest predictors of confidence in the preventive measures and their implementation were the participants' concern of infection and concern of infecting their loved ones, followed by gender and age. Education, settlement size, field and type of employment, household type, own medical problems, and the age and health of the participants' loved ones had a smaller influence on the perceived guidelines importance and implementation. Adherence to measures was positively related to the participants' score in conscientiousness and, in lesser extent, openness. Agreeableness, energy, and emotional stability correlated positively with adherence to basic guidelines. Study provides information useful for developing and adapting future public health policies and interventions., Supplementary Information: The online version contains supplementary material available at 10.1007/s12144-021-02051-5., Competing Interests: Conflicts of Interest/Competing Interests (Include Appropriate Disclosures)The authors declare that they have no conflict of interest., (© The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2021.)

Subterranean ecosystems are among the most widespread environments on Earth, yet we still have poor knowledge of their biodiversity. To raise awareness of subterranean ecosystems, the essential services they provide, and their unique conservation challenges, 2021 and 2022 were designated International Years of Caves and Karst. As these ecosystems have traditionally been overlooked in global conservation agendas and multilateral agreements, a quantitative assessment of solution-based approaches to safeguard subterranean biota and associated habitats is timely. This assessment allows researchers and practitioners to understand the progress made and research needs in subterranean ecology and management. We conducted a systematic review of peer-reviewed and grey literature focused on subterranean ecosystems globally (terrestrial, freshwater, and saltwater systems), to quantify the available evidence-base for the effectiveness of conservation interventions. We selected 708 publications from the years 1964 to 2021 that discussed, recommended, or implemented 1,954 conservation interventions in subterranean ecosystems. We noted a steep increase in the number of studies from the 2000s while, surprisingly, the proportion of studies quantifying the impact of conservation interventions has steadily and significantly decreased in recent years. The effectiveness of 31% of conservation interventions has been tested statistically. We further highlight that 64% of the reported research occurred in the Palearctic and Nearctic biogeographic regions. Assessments of the effectiveness of conservation interventions were heavily biased towards indirect measures (monitoring and risk assessment), a limited sample of organisms (mostly arthropods and bats), and more accessible systems (terrestrial caves). Our results indicate that most conservation science in the field of subterranean biology does not apply a rigorous quantitative approach, resulting in sparse evidence for the effectiveness of interventions. This raises the important question of how to make conservation efforts more feasible to implement, cost-effective, and long-lasting. Although there is no single remedy, we propose a suite of potential solutions to focus our efforts better towards increasing statistical testing and stress the importance of standardising study reporting to facilitate meta-analytical exercises. We also provide a database summarising the available literature, which will help to build quantitative knowledge about interventions likely to yield the greatest impacts depending upon the subterranean species and habitats of interest. We view this as a starting point to shift away from the widespread tendency of recommending conservation interventions based on anecdotal and expert-based information rather than scientific evidence, without quantitatively testing their effectiveness., (© 2022 The Authors. Biological Reviews published by John Wiley & Sons Ltd on behalf of Cambridge Philosophical Society.)

Nitrate (NO 3 - ) pollution is a common phenomenon in karst underground rivers, which are important water sources in karst landscapes. For drinking water safety and environmental protection, it is crucial to accurately identify NO 3 - sources and their migration and transformation processes in the Lihu Underground River. In this study, water samples of the Lihu Underground River in Guangxi were collected in May 2014, October 2014, January 2015, and July 2015, and water chemical and dual isotopic (δ 15 N-NO 3 - and δ 18 O-NO 3 - ) approaches were used to evaluate the NO 3 - characteristics and sources in the Lihu Underground River. The concentration of NO 3 - in the Lihu Underground River ranged from 1.16 to 19.78 mg·L -1 , with an average of 9.30 mg·L -1 , which is more than 37% of the WHO standard (10 mg·L -1 ). The concentrations of NO 3 - in the wet season (May 2014 and July 2015) were slightly lower than those in the dry season (from October 2014 to January 2015) at most sampling sites due to dilution effects. The migration and transformation processes of NO 3 - were analyzed by comparing the measured and calculated concentrations of NO 3 - in the Lihu Underground River. In the dry season (from October 2014 to January 2015), the variation in NO 3 - concentration upstream and midstream of the Lihu Underground River was affected by exogenous input and nitrification. From midstream to the outlet of Xiaolongdong, it is affected by self-purification factors, including physical processes, chemical processes, and biological processes. In the wet season (May 2014 and July 2015), the dilution and mixing effects were the main factors controlling the variation in NO 3 - concentration in the Lihu Underground River. The contribution rates of potential NO 3 - sources (incl. atmospheric precipitation (AP), NO 3 - fertilizer (NF), NH 4 + in fertilizer and rainfall (NFA), soil organic nitrogen (SON), and manure and sewage (M&S)) were quantitatively evaluated by using the IsoSource model. The results showed that in May 2014, the main sources of NO 3 - were M&S and NF, with contribution rates of 46% and 41%, respectively. In October 2014, NO 3 - sources were M&S with a contribution rate of 47%, followed by NFA with a contribution rate of 31%. In January 2015, NO 3 - sources in groundwater were M&S, with a contribution rate of 53%, followed by NFA (34%). In July 2015, the main NO 3 - sources were M&S and NF, whose contribution rates were 54% and 39%, respectively., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Sphagnum mosses are used for biomonitoring air pollution. In 2019, samples were taken from two peat bogs areas in Germany and two in Slovenia to determine differences in their levels of potentially toxic elements (PTE). The Coronavirus Disease (COVID-19) lockdown caused a global decrease in air pollution. Therefore, we repeated the monitoring in 2020 to see if this was also reflected in moss composition. Despite the variability within bogs and the areas, it is possible to distinguish the degree of air pollution between the two countries. In comparison to 2019, the German mosses have higher contents of almost all elements and the Slovenian are enriched in Cr and Hg in 2020. Comparison of the PTE contents, their ratios to Sc, and the enrichment factors show that the COVID-19 lockdown led to a decrease in long-range pollutants bound to finest particles and increased the influence of local soil dusting. The effect prevailed over lower precipitation in 2020 compared to 2019. Transport and industry continued to contribute significantly to contamination. Sphagnum mosses proved to be good indicators of the spatial and temporal extent of pollution. Even relatively short periods of lower air pollution are reflected in moss PTE contents., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Until recently, an extensive area in the central lowlands of the Yucatán peninsula was completely unexplored archaeologically. In 2013 and 2014, during initial surveys in the northern part of the uninhabited Calakmul Biosphere Reserve in eastern Campeche, Mexico, we located Chactún, Tamchén and Lagunita, three major Maya centers with some unexpected characteristics. Lidar data, acquired in 2016 for a larger area of 240 km2, revealed a thoroughly modified and undisturbed archaeological landscape with a remarkably large number of residential clusters and widespread modifications related to water management and agriculture. Substantial additional information was obtained through field surveys and test excavations in 2017 and 2018. While hydraulic and agricultural features and their potential for solving various archaeologically relevant questions were discussed in a previous publication, here we examine the characteristics of settlement patterns, architectural remains, sculpted monuments, and ceramic evidence. The early Middle Preclassic (early first millennium BCE) material collected in stratigraphic pits at Tamchén and another locale constitutes the earliest evidence of colonization known so far in a broader central lowland area. From then until the Late Classic period, which was followed by a dramatic demographic decline, the area under study witnessed relatively constant population growth and interacted with different parts of the Maya Lowlands. However, a number of specific and previously unknown cultural traits attest to a rather distinctive regional development, providing novel information about the extent of regional variation within the Maya culture. By analyzing settlement pattern characteristics, inscriptional data, the distribution of architectural volumes and some other features of the currently visible archaeological landscape, which largely reflects the Late Classic situation, we reconstruct several aspects of sociopolitical and territorial organization in that period, highlighting similarities with and differences from what has been evidenced in the neighboring Río Bec region and elsewhere in the Maya area., Competing Interests: The authors have declared that no competing interests exist.

Historical air temperatures at three karst caves in Slovenia have been compared to current data time series. In Postojnska Jama (PJ), the most visited show cave in Slovenia, the significant temperature difference between historical and modern measurements at the Pulpito site relates to the months April to November. Mean monthly temperatures measured at the Sepolcro site (PJ) in the modern period (2016-2019) are year round significantly higher than in the historical period (1935-1937). The temperature increase over the last 85 years in PJ is attributed to outside temperature rise and additional heat input from visitors, especially for Sepolcro site. A comparison of current (2017-2019) and historical (1956-1957) temperature data in touristically poorly visited Predjama Cave shows lower increase as in PJ and is completely related to outside cave conditions. In the case of Škocjanske Jame (Tiha Jama), air temperature has not significantly increased since the historical 1928 measurements because the monitoring site looks to be morphologically isolated from significant impacts of outside climate and visitors., Supplementary Information: The online version contains supplementary material available at 10.1007/s42452-021-04881-1., Competing Interests: Conflict of interestOn behalf of all authors, the corresponding author states that there is no conflict of interest., (© The Author(s) 2021.)

Relative humidity (RH) was measured at hourly intervals for approximately one year in two caves at seven stations near Playa del Carmen in Quintana Roo, Mexico. Sistema Muévelo Rico is a 1.1 km long cave with 12 entrances and almost no dark zone. Río Secreto (Tuch) is a large river cave with more than 40 km of passages, and an extensive dark zone. Given the need for cave specialists to adapt to saturated humidity, presumably by cuticular thinning, the major stress of RH would be its deviation from saturation. RH in Río Secreto (Tuch) was invariant at three sites and displayed short deviations from 100% RH at the other four sites. These deviations were concentrated at the end of the nortes and beginning of the rainy season. Three of the sites in Sistema Muévelo Rico showed a similar pattern although the timing of the deviations from 100% RH was somewhat displaced. Four sites in Sistema Muévelo Rico were more variable, and were analyzed using a measure of amount of time of deviation from 100% RH for each 24 hour period. Strong seasonality was evident but, remarkably, periods of constant high humidity were not the same at all sites. In most Sistema Muévelo Rico sites, there was a detectable 24 hour cycle in RH, although it was quite weak in about half of them. For Río Secreto (Tuch) only one site showed any sign of a 24 hour cycle. The troglomorphic fauna was more or less uniformly spread throughout the caves and did not concentrate in any one area or set of RH conditions. Compared to temperature, RH is much more constant, perhaps even more constant than the amount of light. However, changes in RH as a result of global warming may have a major negative effect on the subterranean fauna., Competing Interests: MCC Statistical Consulting LLC provided support in the form of salaries for CC. The commercial affiliation of MCC to MCC Statistical Consulting LLC does not alter our adherence to PLOS ONE policies on sharing data and materials.

In arachnids, the Malpighian tubules (MTs), coxal glands and stercoral pockets are capable of collecting and removing excreta from the body. The presence of the MTs among Opiliones was evidenced for the first time in Amilenus aurantiacus in 2015. Individuals undergo a winter diapause subterranean habitats. Here, we provided the morphological and cytological description of the MTs and asked whether their structure and ultrastructure change during the winter diapause. We studied the changes using light and transmission electron microscopy. The MTs consisted of the ureter and a pair of long, lateral blind-ended tubules, forming a long loop in the opisthosoma, and a coiled, terminal ball in the prosoma. The MTs were uniform, composed of a single-cell type, a monolayer of cuboidal epithelial cells, and the basal lamina. The cell ultrastructure was quite comparable to those in other arthropods, except for very long infoldings of the basal membrane protruding close to the nucleus. Except for spherite exploitation, no changes were observed in the ultrastructure of the MT epithelial cells during overwintering. We suggest that the analogous MTs in A. aurantiacus, and the nephron anatomies, along with a single-cell-type MT epithelium, might be of advantage in modelled studies of the nephron., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

When playing in sandboxes, children are particularly vulnerable to poisoning with potentially toxic elements (PTE) due to their hand-to-mouth behaviour. In Slovenia, the city of Celje is heavily contaminated due to zinc ore smelting and iron industry. In some sandboxes, sand from Mežica Pb-Zn mine waste was used. Granulometric and XRF analysis showed that all samples contain larger percentage of fine-sized particles and some have higher As, Ba, Cd, Pb and Zn contents than allowed. By XRD and SEM/EDS analysis we identified carries of PTE, and confirmed three possible sources of contamination: the waste material from the Pb-Zn mine, the emissions from local industry and from the old Zn-smelting stockpile used as a landfill. The total health risk (ΣHI) exceeds critical value in one sandbox. The bioaccessibility of PTEs is alarming due to presence of highly soluble metal-bearing particles. Study revealed importance of combining results of health risk evaluation with bioaccessibility of elements, which is a function of the carriers of PTE. This knowledge is essential for undertaking remedial measures. Improper use of waste material could result in introducing hazardous material in the environment. We propose frequent replacement of sand and stricter legislation regulating status and usage of waste materials., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Two closely related, syntopic species, Leiobunum rupestre and L. subalpinum, spend the day at apparently randomly distributed resting sites. In this preliminary research, we studied differences in their microclimatic preferences at a locality in the Pohorje Mts., Slovenia. We measured air temperature and relative humidity at individual resting sites, and found that L. subalpinum occupied a slightly wider air temperature-relative humidity ecospace as compared to L. rupestre. Individuals of L. rupestre rested at more humid sites than L. subalpinum. Regression analyses revealed that the presence of L. rupestre was most likely at resting sites with air temperature in the range of 1217°C and with relative air humidity of 98.1%. In contrast, the presence of L. subalpinum was most likely at resting sites with air temperature 23.1°C and relative air humidity in the range of 6585%. Nevertheless, microclimatic niches of both species overlapped almost completely, indicating that niche filtering may allow the coexistence of the two species.

The spider major ampullate (MA) silk exhibits high tensile strength and extensibility and is typically a blend of MaSp1 and MaSp2 proteins with the latter comprising glycine-proline-glycine-glycine-X repeating motifs that promote extensibility and supercontraction. The MA silk from Darwin's bark spider ( Caerostris darwini ) is estimated to be two to three times tougher than the MA silk from other spider species. Previous research suggests that a unique MaSp4 protein incorporates proline into a novel glycine-proline-glycine-proline motif and may explain C. darwini MA silk's extraordinary toughness. However, no direct correlation has been made between the silk's molecular structure and its mechanical properties for C. darwini . Here, we correlate the relative protein secondary structure composition of MA silk from C. darwini and four other spider species with mechanical properties before and after supercontraction to understand the effect of the additional MaSp4 protein. Our results demonstrate that C. darwini MA silk possesses a unique protein composition with a lower ratio of helices (31%) and β-sheets (20%) than other species. Before supercontraction, toughness, modulus and tensile strength correlate with percentages of β-sheets, unordered or random coiled regions and β-turns. However, after supercontraction, only modulus and strain at break correlate with percentages of β-sheets and β-turns. Our study highlights that additional information including crystal size and crystal and chain orientation is necessary to build a complete structure-property correlation model.

Heterogeneity in species diversity is driven by the dynamics of speciation and extinction, potentially influenced by organismal and environmental factors. Here, we explore macroevolutionary trends on a phylogeny of golden orbweavers (spider family Nephilidae). Our initial inference detects heterogeneity in speciation and extinction, with accelerated extinction rates in the extremely sexually size dimorphic Nephila and accelerated speciation in Herennia, a lineage defined by highly derived, arboricolous webs, and pronounced island endemism. We evaluate potential drivers of this heterogeneity that relate to organisms and their environment. Primarily, we test two continuous organismal factors for correlation with diversification in nephilids: phenotypic extremeness (female and male body length, and sexual size dimorphism as their ratio) and dispersal propensity (through range sizes as a proxy). We predict a bell-shaped relationship between factor values and speciation, with intermediate phenotypes exhibiting highest diversification rates. Analyses using SSE-class models fail to support our two predictions, suggesting that phenotypic extremeness and dispersal propensity cannot explain patterns of nephilid diversification. Furthermore, two environmental factors (tropical versus subtropical and island versus continental species distribution) indicate only marginal support for higher speciation in the tropics. Although our results may be affected by methodological limitations imposed by a relatively small phylogeny, it seems that the tested organismal and environmental factors play little to no role in nephilid diversification. In the phylogeny of golden orbweavers, the recent hypothesis of universal diversification dynamics may be the simplest explanation of macroevolutionary patterns.

In the 1920s, during the first archaeological excavations at Uaxactún, Petén, Guatemala, an architectural complex named Group E was interpreted as an ancient Maya astronomical observatory, intended specifically for sighting the equinoctial and solstitial sunrises. In the following decades, a large number of architectural compounds with the same configuration have been found, most of them in the central lowlands of the Yucatan peninsula. The multiple hypotheses that have been proposed about the astronomical function of these complexes, commonly designated as E Groups, range from those attributing them a paramount role in astronomical observations to those that consider them merely allegorical or commemorative allusions to celestial cycles, without any observational use. This study, based on quantitative analyses of a reasonably large sample of alignment data, as well as on contextual evidence, shows that many of the previous hypotheses cannot be sustained. I argue that E Groups, although built primarily for ritual purposes, were astronomically functional, but also that they had no specific or particularly prominent role in astronomical observations. Their orientations belong to widespread alignment groups, mostly materialized in buildings of other types and explicable in terms of some fundamental concerns of the agriculturally-based Maya societies. I present the evidence demonstrating that the astronomical orientations initially embedded in E Groups, which represent the earliest standardized form of Maya monumental architecture and whose occurrence in practically all early cities in the central Yucatan peninsula attests to their socio-political significance, were later transferred to buildings and compounds of other types. Therefore, it is precisely the importance of the astronomically and cosmologically significant directions, first incorporated in E Groups, that allows us to understand some prominent aspects of ancient Maya architecture and urbanism., Competing Interests: The author has declared that no competing interests exist.

Hourly temperature was measured for approximately one year at 17 stations in three caves in Quintana Roo, Mexico. Thirteen of these stations were in the extensive twilight zones of all three caves. All seventeen stations showed seasonality in temperature with a 3°C drop during the Nortes season. Two of the caves, Muévelo Sabrosito and Muévelo Rico, showed greater variability during the winter months while in Río Secreto (Tuch) variability was greatest during the rainy season. Río Secreto is less open to the surface than the other two. All sites also showed a daily temperature cycle, although it was very faint in some Río Secreto (Tuch) sites. While temperature variability is diminished relative to surface variation, its temporal pattern is worthy of further study., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: MCC Statistical Consulting LLC provided support in the form of salaries for MCC. The commercial affiliation of MCC to MCC Statistical Consulting LLC does not alter our adherence to PLOS ONE policies on sharing data and materials.

Five decades ago, a landmark paper in Science titled The Cave Environment heralded caves as ideal natural experimental laboratories in which to develop and address general questions in geology, ecology, biogeography, and evolutionary biology. Although the 'caves as laboratory' paradigm has since been advocated by subterranean biologists, there are few examples of studies that successfully translated their results into general principles. The contemporary era of big data, modelling tools, and revolutionary advances in genetics and (meta)genomics provides an opportunity to revisit unresolved questions and challenges, as well as examine promising new avenues of research in subterranean biology. Accordingly, we have developed a roadmap to guide future research endeavours in subterranean biology by adapting a well-established methodology of 'horizon scanning' to identify the highest priority research questions across six subject areas. Based on the expert opinion of 30 scientists from around the globe with complementary expertise and of different academic ages, we assembled an initial list of 258 fundamental questions concentrating on macroecology and microbial ecology, adaptation, evolution, and conservation. Subsequently, through online surveys, 130 subterranean biologists with various backgrounds assisted us in reducing our list to 50 top-priority questions. These research questions are broad in scope and ready to be addressed in the next decade. We believe this exercise will stimulate research towards a deeper understanding of subterranean biology and foster hypothesis-driven studies likely to resonate broadly from the traditional boundaries of this field., (© 2020 Cambridge Philosophical Society.)

Vicariance and dispersal events, combined with intricate global climatic history, have left an imprint on the spatiotemporal distribution and diversity of many organisms. Anelosimus cobweb spiders (Theridiidae), are organisms ranging in behavior from solitary to highly social, with a cosmopolitan distribution in temperate to tropical areas. Their evolutionary history and the discontinuous distribution of species richness suggest that 1) long-distance overwater dispersal and 2) climate change during the Neogene (23-2.6 Ma), may be major factors in explaining their distribution and diversification. Here, we test these hypotheses, and explicitly test if global Miocene/Pliocene climatic cooling in the last 8 Ma affected Anelosimus radiation in parallel in South America and Madagascar. To do so, we investigate the phylogeny and spatiotemporal biogeography of Anelosimus through a culmination of a 20-year comprehensive global sampling at the species level (69 species, including 84% of the known 75 species worldwide, represented by 268 individuals) using nucleotide data from seven loci (5.5 kb). Our results strongly support the monophyly of Anelosimus with an Oligocene ($\sim $30 Ma) South American origin. Major clades on other continents originate via multiple, long-distance dispersal events, of solitary or subsocial-but not social-lineages, from the Americas. These intercontinental dispersals were to Africa, Madagascar (twice), and SE Asia/Australasia. The early diversification of Anelosimus spiders coincides with a sudden thermal increase in the late Oligocene ($\sim $27-25 Ma), though no causal connection can be made. Our results, however, strongly support the hypothesis that global Neogene climatic cooling in the last 8 Ma drove Anelosimus radiation in parallel in South America and Madagascar, offering a rare empirical evidence for diversification of a socially diverse group driven by an interplay between long-distance dispersal and global Neogene climatic changes. [Cobweb spiders; diversification; global biogeography; long-distance dispersal; molecular phylogenetics; neogene climate changes; sociality; vicariance.]., (© The Author(s) 2020. Published by Oxford University Press, on behalf of the Society of Systematic Biologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Determining species boundaries forms an important foundation for biological research. However, the results of molecular species delimitation can vary with the data sets and methods that are used. Here we use a two-step approach to delimit species in the genus Heptathela, a group of primitively segmented trapdoor spiders that are endemic to Japanese islands. Morphological evidence suggests the existence of 19 species in the genus. We tested this initial species hypothesis by using six molecular species-delimitation methods to analyse 180 mitochondrial COI sequences of Heptathela sampled from across the known range of the genus. We then conducted a set of more focused analyses by sampling additional genetic markers from the subset of taxa that were inconsistently delimited by the single-locus analyses of mitochondrial DNA. Multilocus species delimitation was performed using two Bayesian approaches based on the multispecies coalescent. Our approach identified 20 putative species among the 180 sampled individuals of Heptathela. We suggest that our two-step approach provides an efficient strategy for delimiting species while minimizing costs and computational time., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Broomcorn millet (Panicum miliaceum L.) is not one of the founder crops domesticated in Southwest Asia in the early Holocene, but was domesticated in northeast China by 6000 BC. In Europe, millet was reported in Early Neolithic contexts formed by 6000 BC, but recent radiocarbon dating of a dozen 'early' grains cast doubt on these claims. Archaeobotanical evidence reveals that millet was common in Europe from the 2nd millennium BC, when major societal and economic transformations took place in the Bronze Age. We conducted an extensive programme of AMS-dating of charred broomcorn millet grains from 75 prehistoric sites in Europe. Our Bayesian model reveals that millet cultivation began in Europe at the earliest during the sixteenth century BC, and spread rapidly during the fifteenth/fourteenth centuries BC. Broomcorn millet succeeds in exceptionally wide range of growing conditions and completes its lifecycle in less than three summer months. Offering an additional harvest and thus surplus food/fodder, it likely was a transformative innovation in European prehistoric agriculture previously based mainly on (winter) cropping of wheat and barley. We provide a new, high-resolution chronological framework for this key agricultural development that likely contributed to far-reaching changes in lifestyle in late 2nd millennium BC Europe.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Local biodiversity trends over time are likely to be decoupled from global trends, as local processes may compensate or counteract global change. We analyze 161 long-term biological time series (15-91 years) collected across Europe, using a comprehensive dataset comprising ~6,200 marine, freshwater and terrestrial taxa. We test whether (i) local long-term biodiversity trends are consistent among biogeoregions, realms and taxonomic groups, and (ii) changes in biodiversity correlate with regional climate and local conditions. Our results reveal that local trends of abundance, richness and diversity differ among biogeoregions, realms and taxonomic groups, demonstrating that biodiversity changes at local scale are often complex and cannot be easily generalized. However, we find increases in richness and abundance with increasing temperature and naturalness as well as a clear spatial pattern in changes in community composition (i.e. temporal taxonomic turnover) in most biogeoregions of Northern and Eastern Europe.

Karst aquifers provide drinking water for 10% of the world's population, support agriculture, groundwater-dependent activities, and ecosystems. These aquifers are characterised by complex groundwater-flow systems, hence, they are extremely vulnerable and protecting them requires an in-depth understanding of the systems. Poor data accessibility has limited advances in karst research and realistic representation of karst processes in large-scale hydrological studies. In this study, we present World Karst Spring hydrograph (WoKaS) database, a community-wide effort to improve data accessibility. WoKaS is the first global karst springs discharge database with over 400 spring observations collected from articles, hydrological databases and researchers. The dataset's coverage compares to the global distribution of carbonate rocks with some bias towards the latitudes of more developed countries. WoKaS database will ensure easy access to a large-sample of good quality datasets suitable for a wide range of applications: comparative studies, trend analysis and model evaluation. This database will largely contribute to research advancement in karst hydrology, supports karst groundwater management, and promotes international and interdisciplinary collaborations.

Species occupying habitats subjected to frequent natural and/or anthropogenic changes are a challenge for conservation management. We studied one such species, Viola uliginosa , an endangered perennial wetland species typically inhabiting sporadically flooded meadows alongside rivers/lakes. In order to estimate genomic diversity, population structure, and history, we sampled five sites in Finland, three in Estonia, and one each in Slovenia, Belarus, and Poland using genomic SNP data with double-digest restriction site-associated DNA sequencing (ddRAD-seq). We found monophyletic populations, high levels of inbreeding (mean population F SNP  = 0.407-0.945), low effective population sizes ( N e  = 0.8-50.9), indications of past demographic expansion, and rare long-distance dispersal. Our results are important in implementing conservation strategies for V. uliginosa , which should include founding of seed banks, ex situ cultivations, and reintroductions with individuals of proper origin, combined with continuous population monitoring and habitat management., Competing Interests: None declared., (© 2020 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Sexual size dimorphism is one of the most striking animal traits, and among terrestrial animals, it is most extreme in certain spider lineages. The most extreme sexual size dimorphism (eSSD) is female biased. eSSD itself is probably an epiphenomenon of gendered evolutionary drivers whose strengths and directions are diverse. We demonstrate that eSSD spider clades are aberrant by sampling randomly across all spiders to establish overall averages for female (6.9 mm) and male (5.6 mm) size. At least 16 spider eSSD clades exist. We explore why the literature does not converge on an overall explanation for eSSD and propose an equilibrium model featuring clade- and context-specific drivers of gender size variation. eSSD affects other traits such as sexual cannibalism, genital damage, emasculation, and monogyny with terminal investment. Coevolution with these extreme sexual phenotypes is termed eSSD mating syndrome. Finally, as costs of female gigantism increase with size, eSSD may represent an evolutionary dead end.

Distributed environmental research infrastructures are important to support assessments of the effects of global change on landscapes, ecosystems and society. These infrastructures need to provide continuity to address long-term change, yet be flexible enough to respond to rapid societal and technological developments that modify research priorities. We used a horizon scanning exercise to identify and prioritize emerging research questions for the future development of ecosystem and socio-ecological research infrastructures in Europe. Twenty research questions covered topics related to (i) ecosystem structures and processes, (ii) the impacts of anthropogenic drivers on ecosystems, (iii) ecosystem services and socio-ecological systems and (iv), methods and research infrastructures. Several key priorities for the development of research infrastructures emerged. Addressing complex environmental issues requires the adoption of a whole-system approach, achieved through integration of biotic, abiotic and socio-economic measurements. Interoperability among different research infrastructures needs to be improved by developing standard measurements, harmonizing methods, and establishing capacities and tools for data integration, processing, storage and analysis. Future research infrastructures should support a range of methodological approaches including observation, experiments and modelling. They should also have flexibility to respond to new requirements, for example by adjusting the spatio-temporal design of measurements. When new methods are introduced, compatibility with important long-term data series must be ensured. Finally, indicators, tools, and transdisciplinary approaches to identify, quantify and value ecosystem services across spatial scales and domains need to be advanced., (Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Among the eight extant genera of primitively segmented spiders, family Liphistiidae, two are confined to East Asian islands, Heptathela Kishida, 1923 and Ryuthela Haupt, 1983. In this paper, a taxonomic revision of the genus Heptathela (Heptathelinae) from Kyushu and Ryukyu archipelago, Japan is provided. This study follows a multi-tier species delimitation strategy within an integrative taxonomic framework that is presented in a parallel paper, in which diagnosable lineages are considered as valid species. There, the initial hypothesis of species diversity (19) based on classical morphological diagnoses is tested with multiple species delimitation methods aimed at resolving conflict in data. This revision follows those analyses that converge on the species diversity of 20, which includes a pair of cryptic species that would have been undetected with morphology alone. After this revision, eight previously described species remain valid, two junior synonyms are proposed, and 12 new Heptathela species are described based on diagnostic evidence. To ease identification and to hint at putative evolutionary units, Heptathela is divided into three groups. The Kyushu group contains H. higoensis Haupt, 1983, H. kikuyai Ono, 1998, H. kimurai (Kishida, 1920), and H. yakushimaensis Ono, 1998; the Amami group contains H. amamiensis Haupt, 1983, H. kanenoi Ono, 1996, H. kojima sp. nov. , H. sumiyo sp. nov. , and H. uken sp. nov. ; and the Okinawa group contains H. yanbaruensis Haupt, 1983, H. aha sp. nov. , H. gayozan sp. nov. , H. kubayama sp. nov. , H. mae sp. nov. , H. otoha sp. nov. , H. shuri sp. nov. , H. tokashiki sp. nov. , H. unten sp. nov. , and H. crypta sp. nov. Heptathela helios Tanikawa & Miyashita, 2014 is not assigned to a species group. A combination of diagnostic tools augments the morphological diagnoses that, in isolation, would be prone to error in morphologically challenging groups of organisms., (Xin Xu, Hirotsugu Ono, Matjaž Kuntner, Fengxiang Liu, Daiqin Li.)

Psychiatric re-hospitalisation rates have been of longstanding interest as health care quality metric for planners and policy makers, but are criticized for not being comparable across hospitals and countries due to measurement unclarities. The objectives of the present study were to explore the interoperability of national electronic routine health care registries of six European countries (Austria, Finland, Italy, Norway, Romania, Slovenia) and, by using variables found to be comparable, to calculate and compare re-hospitalisation rates and the associated risk factors. A "Methods Toolkit" was developed for exploring the interoperability of registry data and protocol led pilot studies were carried out. Problems encountered in this process are described. Using restricted but comparable data sets, up to twofold differences in psychiatric re-hospitalisation rates were found between countries for both a 30- and 365-day follow-up period. Cumulative incidence curves revealed noteworthy additional differences. Health system characteristics are discussed as potential causes for the differences. Multi-level logistic regression analyses showed that younger age and a diagnosis of schizophrenia/mania/bipolar disorder consistently increased the probability of psychiatric re-hospitalisation across countries. It is concluded that the advantage of having large unselected study populations of national electronic health care registries needs to be balanced against the considerable efforts to examine the interoperability of databases in cross-country comparisons., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Current climate change has led to latitudinal and altitudinal range expansions of numerous species. During such range expansions, plant species are expected to experience changes in interactions with other organisms, especially with belowground biota that have a limited dispersal capacity. Nematodes form a key component of the belowground food web as they include bacterivores, fungivores, omnivores and root herbivores. However, their community composition under climate change-driven intracontinental range-expanding plants has been studied almost exclusively under controlled conditions, whereas little is known about actual patterns in the field. Here, we use novel molecular sequencing techniques combined with morphological quantification in order to examine nematode communities in the rhizospheres of four range-expanding and four congeneric native species along a 2,000 km latitudinal transect from South-Eastern to North-Western Europe. We tested the hypotheses that latitudinal shifts in nematode community composition are stronger in range-expanding plant species than in congeneric natives and that in their new range, range-expanding plant species accumulate fewest root-feeding nematodes. Our results show latitudinal variation in nematode community composition of both range expanders and native plant species, while operational taxonomic unit richness remained the same across ranges. Therefore, range-expanding plant species face different nematode communities at higher latitudes, but this is also the case for widespread native plant species. Only one of the four range-expanding plant species showed a stronger shift in nematode community composition than its congeneric native and accumulated fewer root-feeding nematodes in its new range. We conclude that variation in nematode community composition with increasing latitude occurs for both range-expanding and native plant species and that some range-expanding plant species may become released from root-feeding nematodes in the new range., (© 2019 The Authors Global Change Biology Published by John Wiley & Sons Ltd.)

Darwin's bark spider ( Caerostris darwini ) produces giant orb webs from dragline silk that can be twice as tough as other silks, making it the toughest biological material. This extreme toughness comes from increased extensibility relative to other draglines. We show C. darwini dragline-producing major ampullate (MA) glands highly express a novel silk gene transcript (MaSp4) encoding a protein that diverges markedly from closely related proteins and contains abundant proline, known to confer silk extensibility, in a unique GPGPQ amino acid motif. This suggests C. darwini evolved distinct proteins that may have increased its dragline's toughness, enabling giant webs. Caerostris darwini's MA spinning ducts also appear unusually long, potentially facilitating alignment of silk proteins into extremely tough fibers. Thus, a suite of novel traits from the level of genes to spinning physiology to silk biomechanics are associated with the unique ecology of Darwin's bark spider, presenting innovative designs for engineering biomaterials., Competing Interests: Competing interestsThe authors declare no competing interests.

Our understanding of spider reproductive biology is hampered by the vast anatomical diversity and difficulties associated with its study. Although authors agree on the two general types of female spider genitalia, haplogyne (plesiomorphic) and entelegyne (apomorphic), our understanding of variation within each group mostly concerns the external genital part, while the internal connections with the reproductive duct are largely unknown. Conventionally and simplistically, the spermathecae of haplogynes have simple two-way ducts, and those of entelegynes have separate copulatory and fertilization ducts for sperm to be transferred in and out of spermathecae, respectively. Sperm is discharged from the spermathecae directly into the uterus externus (a distal extension of the oviduct), which, commonly thought as homologous in both groups, is the purported location of internal fertilization in spiders. However, the structural evolution from haplo- to entelegyny remains unresolved, and thus the precise fertilization site in entelegynes is ambiguous. We aim to clarify this anatomical problem through a widely comparative morphological study of internal female genital system in entelegynes. Our survey of 147 epigyna (121 examined species in 97 genera, 34 families) surprisingly finds no direct connection between the fertilization ducts and the uterus externus, which, based on the homology with basal-most spider lineages, is a dead-end caecum in entelegynes. Instead, fertilization ducts usually connect with a secondary uterus externus, a novel feature taking over the functional role of the plesiomorphic uterus externus. We hypothesize that the transition from haplo- to entelegyny entailed not only the emergence of the two separate duct systems (copulatory, fertilization), but also involved substantial morphological changes in the distal part of the oviduct. Thus, the common oviduct may have shifted its distal connection from the uterus externus to the secondary uterus externus, perhaps facilitating discharge of larger eggs. Our findings suggest that the conventional model of entelegyne reproduction needs redefinition., Competing Interests: The authors have declared that no competing interests exist.

Instances of sexual size dimorphism (SSD) provide the context for rigorous tests of biological rules of size evolution, such as Cope's rule (phyletic size increase), Rensch's rule (allometric patterns of male and female size), as well as male and female body size optima. In certain spider groups, such as the golden orbweavers (Nephilidae), extreme female-biased SSD (eSSD, female:male body length $\ge$2) is the norm. Nephilid genera construct webs of exaggerated proportions, which can be aerial, arboricolous, or intermediate (hybrid). First, we established the backbone phylogeny of Nephilidae using 367 anchored hybrid enrichment markers, then combined these data with classical markers for a reference species-level phylogeny. Second, we used the phylogeny to test Cope and Rensch's rules, sex specific size optima, and the coevolution of web size, type, and features with female and male body size and their ratio, SSD. Male, but not female, size increases significantly over time, and refutes Cope's rule. Allometric analyses reject the converse, Rensch's rule. Male and female body sizes are uncorrelated. Female size evolution is random, but males evolve toward an optimum size (3.2-4.9 mm). Overall, female body size correlates positively with absolute web size. However, intermediate sized females build the largest webs (of the hybrid type), giant female Nephila and Trichonephila build smaller webs (of the aerial type), and the smallest females build the smallest webs (of the arboricolous type). We propose taxonomic changes based on the criteria of clade age, monophyly and exclusivity, classification information content, and diagnosability. Spider families, as currently defined, tend to be between 37 million years old and 98 million years old, and Nephilidae is estimated at 133 Ma (97-146), thus deserving family status. We, therefore, resurrect the family Nephilidae Simon 1894 that contains Clitaetra Simon 1889, the Cretaceous GeratonephilaPoinar and Buckley (2012), Herennia Thorell 1877, IndoetraKuntner 2006, new rank, Nephila Leach 1815, Nephilengys L. Koch 1872, Nephilingis Kuntner 2013, Palaeonephila Wunderlich 2004 from Tertiary Baltic amber, and TrichonephilaDahl 1911, new rank. We propose the new clade Orbipurae to contain Araneidae Clerck 1757, Phonognathidae Simon 1894, new rank, and Nephilidae. Nephilid female gigantism is a phylogenetically ancient phenotype (over 100 Ma), as is eSSD, though their magnitudes vary by lineage., (© The Author(s) 2018. Published by Oxford University Press on behalf of the Society of Systematic Biologists.)

The European cave spider, Meta menardi, is a representative of the troglophiles, i.e. non-strictly subterranean organisms. Our aim was to interpret the cytological results from an ecological perspective, and provide a synthesis of the hitherto knowledge about M. menardi into a theory of key features marking it a troglophile. We studied ultrastructural changes of the midgut epithelial cells in individuals spending winter under natural conditions in caves, using light microscopy and TEM. The midgut diverticula epithelium consisted of secretory cells, digestive cells and adipocytes. During winter, gradual vacuolization of some digestive cells appeared, and some necrotic digestive cells and necrotic adipocytes appeared. This cytological information completes previous studies on M. menardi starved under controlled conditions in the laboratory. In experimental starvation and natural winter conditions, M. menardi gradually exploit reserve compounds from spherites, protein granules and through autophagy, and energy-supplying lipids and glycogen, as do many overwintering arthropods. We found no special cellular response to living in the habitat. Features that make it partly adapted to the subterranean habitat include starvation hardiness as a possible preadaptation, an extremely opportunistic diet, a partly reduced orb, tracking and capturing prey on bare walls and partly reduced tolerance to below-zero temperatures.

Selection pressures leading to extreme, female-biased sexual size dimorphism (SSD) in spiders continue to be debated. It has been proposed that males of sexually size dimorphic spiders could be small because gravity constrains adult agility (locomotor abilities). Accordingly, small males should achieve higher vertical climbing speeds and should be more prone to bridge. The curvilinear model of the gravity hypothesis predicts a negative relationship between vertical climbing speed and male body size only over a threshold of 7.6 mm, 42.5 mg. Because males of most species with extreme SSD fall well below this threshold, the relationship between male size and agility at this scale remains vague. Here, we tested three hypotheses on how male size, mass and age (after maturation) relate to vertical climbing and bridging ability in Nephilingis cruentata, a highly sexually dimorphic orb-weaver with males well below the size threshold. We placed males of different sizes and adult ages in a vertical platform and recorded their climbing speeds. Contrary to the original study testing male bridging ability as binary variable, we measured the duration of the crossing of the bridging thread, as well as its sagging distance. Male body size and mass positively related to the vertical climbing speed and to the distance of the sagging thread during bridging, but had no influence on the bridging duration. The detected positive correlation between male size/mass and vertical climbing speed goes against our first prediction, that small males would have vertical climbing advantage in Nephilingis cruentata, but agrees with the curvilinear model. Against our second prediction, small males were not faster during bridging. Finally, in agreement with our third prediction, threads sagged more in heavier males. These results suggest that small male size confers no agility advantages in Nephilingis cruentata., Competing Interests: The authors have declared that no competing interests exist.