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Male orchid bees are unique in the animal kingdom for making perfumes that function as sex pheromone. Males collect volatile chemicals from the environment in the neotropical forests, including floral and non-floral sources, creating complex but species-specific blends. Male orchid bees exhibit several adaptations to facilitate perfume collection and storage. When collecting volatile compounds, males apply lipid substances that they secrete from cephalic labial glands onto the fragrant substrate. These lipids help dissolve and retain the volatiles, similar to the process of 'enfleurage' in the traditional perfume industry. We investigated how the chemical composition of acquired perfume and labial gland secretions varied across the phylogeny of orchid bees, including 65 species in five genera from Central and South America. Perfumes showed rapid evolution as revealed by low overall phylogenetic signal, in agreement with the idea that perfume compounds diverge rapidly and substantially among closely related species due to their role in species recognition. A possible exception were perfumes in the genus Eulaema, clustering closely in chemospace, partly mediated by high proportions of carvone and trans-carvone oxide. Labial gland secretions, in contrast, showed a strong phylogenetic signal at the genus level, with secretions of Eufriesea and Exaerete dominated by fatty acids and Eulaema dominated by saturated acetates of chain lengths 12 to 16 C-atoms. Secretions of the majority of Euglossa were heavily dominated by one unsaturated long chain diacetate, (9Z)-Eicosen-1,20-diyldiacetate. However, we also identified few highly divergent species of Euglossa in four subclades (11 species) that appear to have secondarily replaced the diacetate with other compounds. In comparison with environment-derived perfumes, the evolution of labial gland secretion is much slower, likely constrained by the underlying biochemical pathways, but perhaps influenced by perfume-solvent chemical interactions., (© 2024. The Author(s).)

A multitude of anthropogenic stressors impact biological communities and ecosystem processes in urban streams. Prominent among them are salinization, increased temperature, and altered flow regimes, all of which can affect microbial decomposer communities and litter decomposition, a fundamental ecosystem process in streams. Impairments caused by these stressors individually or in combination and recovery of communities and ecosystem processes after release from these stressors are not well understood. To improve our understanding of multiple stressors impacts we performed an outdoor stream mesocosm experiment with 64 experimental units to assess the response of microbial litter decomposers and decomposition. The three stressors we applied in a full-factorial design were increased salinity (NaCl addition, 0.53 mS cm -1 above ambient), elevated temperature (3.5 °C above ambient), and reduced flow velocity (3.5 vs 14.2 cm s -1 ). After two weeks of stressor exposure (first sampling) and two subsequent weeks of recovery (second sampling), we determined leaf-associated microbial respiration, fungal biomass, and the sporulation activity and community composition of aquatic hyphomycetes in addition to decomposition rates of black alder (Alnus glutinosa) leaves confined in fine-mesh litter bags. Microbial colonization of the litter was accompanied by significant mass loss in all mesocosms. However, there was little indication that mass loss, microbial respiration, fungal biomass, sporulation rate or community composition of aquatic hyphomycetes was strongly affected by either single stressors or their interactions. Two exceptions were temperature effects on sporulation and decomposition rate. Similarly, no notable differences among mesocosms were observed after the recovery phase. These results suggest that microbial decomposers and leaf litter decomposition are either barely impaired by exposure to the tested stressors at the levels applied in our experiment, or that communities in restored urban streams are well adapted to cope with these stressor levels., 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 The Authors. Published by Elsevier B.V. All rights reserved.)

Field observations form the basis of the majority of studies on microphytobenthic algal communities in freshwater ecosystems. Controlled mesocosm experiments data are comparatively uncommon. The few experimental mesocosm studies that have been conducted provide valuable insights into how multiple stressors affect the community structures and photosynthesis-related traits of benthic microalgae. The recovery process after the stressors have subsided, however, has received less attention in mesocosm studies. To close this gap, here we present the results of a riparian mesocosm experiment designed to investigate the effects of reduced flow velocity, increased salinity and increased temperature on microphytobenthic communities. We used a full factorial design with a semi-randomised distribution of treatments consisting of two levels of each stressor (2 × 2 × 2 treatments), with eight replicates making a total of 64 circular mesocosms, allowing a nuanced examination of their individual and combined influences. We aimed to elucidate the responses of microalgae communities seeded from stream water to the applied environmental stressors. Our results showed significant effects of reduced flow velocity and increased temperature on microphytobenthic communities. Recovery after stressor treatment led to a convergence in community composition, with priority effects (hypothesized to reflect competition for substrate between resident and newly arriving immigrant taxa) slowing down community shifts and biomass increase. Our study contributes to the growing body of literature on the ecological dynamics of microphytobenthos and emphasises the importance of rigorous experiments to validate hypotheses. These results encourage further investigation into the nuanced interactions between microphytobenthos and their environment and shed light on the complexity of ecological responses in benthic systems., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Hygroscopic seed-scale movement is responsible for the weather-adaptive opening and closing of pine cones and for facilitating seed dispersal under favorable environmental conditions. Although this phenomenon has long been investigated, many involved processes are still not fully understood. To gain a deeper mechanical and structural understanding of the cone and its functional units, namely the individual seed scales, we have investigated their desiccation- and wetting-induced movement processes in a series of analyses and manipulative experiments. We found, for example, that the abaxial scale surface is responsible for the evaporation of water from the closed cone and subsequent cone opening. Furthermore, we tested the capability of dry and deformed scales to restore their original shape and biomechanical properties by wetting. These results shed new light on the orchestration of scale movement in cones and the involved forces and provide information about the functional robustness and resilience of cones, leading to a better understanding of the mechanisms behind hygroscopic pine cone opening, the respective ecological framework, and, possibly, to the development of smart biomimetic actuators.

Urban streams are exposed to a variety of anthropogenic stressors. Freshwater salinization is a key stressor in these ecosystems that is predicted to be further exacerbated by climate change, which causes simultaneous changes in flow parameters, potentially resulting in non-additive effects on aquatic ecosystems. However, the effects of salinization and flow velocity on urban streams are still poorly understood as multiple-stressor experiments are often conducted at pristine rather than urban sites. Therefore, we conducted a mesocosm experiment at the Boye River, a recently restored stream located in a highly urbanized area in Western Germany, and applied recurrent pulses of salinity along a gradient (NaCl, 9 h daily of +0 to +2.5 mS/cm) in combination with normal and reduced current velocities (20 cm/s vs. 10 cm/s). Using a comprehensive assessment across multiple organism groups (macroinvertebrates, eukaryotic algae, fungi, parasites) and ecosystem functions (primary production, organic-matter decomposition), we show that flow velocity reduction has a pervasive impact, causing community shifts for almost all assessed organism groups (except fungi) and inhibiting organic-matter decomposition. Salinization affected only dynamic components of community assembly by enhancing invertebrate emigration via drift and reducing fungal reproduction. We caution that the comparatively small impact of salt in our study can be due to legacy effects from past salt pollution by coal mining activities >30 years ago. Nevertheless, our results suggest that urban stream management should prioritize the continuity of a minimum discharge to maintain ecosystem integrity. Our study exemplifies a holistic approach for the assessment of multiple-stressor impacts on streams, which is needed to inform the establishment of a salinity threshold above which mitigation actions must be taken., 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 The Authors. Published by Elsevier B.V. All rights reserved.)

Male neotropical orchid bees concoct complex perfume blends by collecting exogenous volatiles from various sources, including orchids. These perfumes, stored in specialized hind-leg pouches and released during courtship, serve as inter-sexual signals. It has been hypothesized that male perfumes honestly indicate aspects of male fitness. If perfume traits such as quantity or complexity increase over individual lifetime, perfumes could reflect age (survival) and cumulative foraging success of males. We conducted a two-season mark-recapture study with Euglossa imperialis in Costa Rica, monitoring the balance of perfume uptake and expenditure over individual male lifetime. We sealed one hind-leg pouch upon initial capture, 'freezing' the perfume status on one side, and compared it with the other side at recapture to assess changes in perfume traits over time. Additionally, we used a novel method to estimate individual age by combining two parameters of wing degradation. Contrary to predictions, young to intermediate-aged bees had the highest quantities of perfume and the highest diversity of detected compounds. At the same time, the change in perfume between recaptures was positive (increase in amount and complexity) in young bees, whereas it was neutral to negative in older bees. Although these findings do not disprove an indicator function of male perfume, they shift the emphasis to non-cumulative fitness components such as sensory acuteness or cognitive capacity as likely targets of selection. Females preferring strong perfume signals in mates would maximize speed of foraging in offspring rather than their lifetime cumulative yield., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2024. Published by The Company of Biologists Ltd.)

Perfume making in male orchid bees is a unique behavior that has given rise to an entire pollination syndrome in the neotropics. 1 Male orchid bees concoct and store species-specific perfume mixtures in specialized hind-leg pockets , 2 However, the function and the ultimate causes of this behavior have remained elusive. 3 using volatiles acquired from multiple environmental sources, including orchid flowers. 4 Here, we demonstrate that the possession of perfume increases male mating success and paternity in Euglossa dilemma, a species of orchid bees recently naturalized in Florida. We supplemented males reared from trap-nests with perfume loads harvested from wild conspecifics. In dual-choice experiments, males supplemented with perfumes mated with more females, and sired more offspring, than untreated, equal-aged, control males. Although perfume supplementation had little effect on the intensity of male courtship display, it changed the dynamics of male-male interactions. Our results demonstrate that male-acquired perfumes are sexual signals that stimulate females for mating and suggest that sexual selection is key in shaping the evolution of perfume communication in orchid bees., 5 However, the function and the ultimate causes of this behavior have remained elusive. 2 , 6 Although previous observations suggested that male perfumes serve as chemical signals, the attractiveness for females has not be shown. 7 , 8 Here, we demonstrate that the possession of perfume increases male mating success and paternity in Euglossa dilemma, a species of orchid bees recently naturalized in Florida. We supplemented males reared from trap-nests with perfume loads harvested from wild conspecifics. In dual-choice experiments, males supplemented with perfumes mated with more females, and sired more offspring, than untreated, equal-aged, control males. Although perfume supplementation had little effect on the intensity of male courtship display, it changed the dynamics of male-male interactions. Our results demonstrate that male-acquired perfumes are sexual signals that stimulate females for mating and suggest that sexual selection is key in shaping the evolution of perfume communication in orchid bees., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Females of frog-biting midges (Corethrellidae) obtain their blood meals from male calling frogs. While the morphology of the feeding apparatus is well studied in hematophagous Diptera that impact humans, frog-biting midges have received far less attention. We provide a detailed micromorphological examination of the piercing blood-sucking proboscis and maxillary palpus in three Corethrella species using scanning electron microscopy and histological semi-thin sectioning. We also compare the sensilla found on the proboscis tip and the palpus of Corethrella with other piercing blood-sucking Diptera. Corethrella spp. have a proboscis length of about 135 µm, equipped with delicate mandibular piercing structures composing the food canal together with the labrum and hypopharynx. Their proboscis composition is plesiomorphic and more similar to other short-proboscid hematophagous Culicomorpha (e.g., Simuliidae), in contrast to the phylogenetically more closely related long-proboscid Culicidae. As in other short-proboscid taxa, the salivary canal in Corethrella spp. transitions into an open salivary groove with one mandible forming a seal, whereas in Culicidae the salivary canal is closed until the tip of the proboscis. We discuss the possible functional constraints of very short, piercing blood-sucking proboscises (e.g., dimensions of host blood cells) that may limit the size of the food canal.

Vector-borne pathogens exist in obligate transmission cycles between vector and reservoir host species. Host and vector shifts can lead to geographic expansion of infectious agents and the emergence of new diseases in susceptible individuals. Three bacterial genospecies (Borrelia afzelii, Borrelia bavariensis, and Borrelia garinii) predominantly utilize two distinct tick species as vectors in Asia (Ixodes persulcatus) and Europe (Ixodes ricinus). Through these vectors, the bacteria can infect various vertebrate groups (e.g., rodents, birds) including humans where they cause Lyme borreliosis, the most common vector-borne disease in the Northern hemisphere. Yet, how and in which order the three Borrelia genospecies colonized each continent remains unclear including the evolutionary consequences of this geographic expansion. Here, by reconstructing the evolutionary history of 142 Eurasian isolates, we found evidence that the ancestors of each of the three genospecies probably have an Asian origin. Even so, each genospecies studied displayed a unique substructuring and evolutionary response to the colonization of Europe. The pattern of allele sharing between continents is consistent with the dispersal rate of the respective vertebrate hosts, supporting the concept that adaptation of Borrelia genospecies to the host is important for pathogen dispersal. Our results highlight that Eurasian Lyme borreliosis agents are all capable of geographic expansion with host association influencing their dispersal; further displaying the importance of host and vector association to the geographic expansion of vector-borne pathogens and potentially conditioning their capacity as emergent pathogens., (© 2022 The Authors. Molecular Ecology published by John Wiley & Sons Ltd.)

Fluoxetine, a common pharmaceutical used as an antidepressant, is already considered potentially hazardous to biota due to its increasing use and detection in European, North American, and Asian rivers. We studied the effects of fluoxetine on Daphnia magna, as we hypothesized that fluoxetine might have harmful effects, short and long-term, at different levels: survival, behaviour, and reproduction (offspring production). We applied two different approaches: (i) a scenario at environmentally relevant concentrations (0.1-1.0 μg/L) and (ii) a scenario simulating a future worsening of contamination (1-800 μg/L) until the reach of lethal concentrations. In the former, we examined whether there are multigenerational effects on reproduction and on the avoidance/colonisation behaviour in previously exposed populations. In the latter, three responses were assessed: survival, avoidance behaviour and reproduction. We did not detect differences in the reproduction output of D. magna among the treatments over the three generations examined. Irrespective of the multigenerational treatment, D. magna colonised the environments with fluoxetine in a similar way. In the second scenario, we determined the lethal concentration for 50% of the population (96 h-LC 50  = 365 μg/L), which, in spite of the toxic effect, was attractive to organisms during the avoidance tests (24 h); in fact, D. magna were attracted (no repellence) even to the highest concentrations of fluoxetine tested (800 μg/L). Lastly, in a 21-day chronic toxicity test the reproduction output of D. magna increased with higher concentrations of fluoxetine. This effect might be related to the fact that the organisms in the contaminated treatment began their first reproduction earlier, when compared to that in the control treatments. In conclusion, this study discusses an identified hazard for aquatic biota due to the fluoxetine attraction effect and a predictive assessment of the consequences expected if its indiscriminate use increases., 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 © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Pine cones show functionally highly resilient, hygroscopically actuated opening and closing movements, which are repeatable and function even in millions of years old, coalified cones. Although the functional morphology and biomechanics behind the individual seed scale motions are well understood, the initial opening of the cone, which is often accompanied by an audible cracking noise, is not. We therefore investigated the initial opening events of mature fresh cones of Scots pine ( Pinus sylvestris ) and their subsequent motion patterns. Using high-speed and time lapse videography, 3D digital image correlation techniques, force measurements, thermographic and chemical-rheological resin analyses, we are able to draw a holistic picture of the initial opening process involving the rupture of resin seals and very fast seed scale motion in the millisecond regime. The rapid cone opening was not accompanied by immediate seed release in our experiments and, therefore, cannot be assigned to ballistochory. As the involved passive hydraulic-elastic processes in cracking are very fine-tuned, we hypothesize that they are under tight mechanical-structural control to ensure an ecologically optimized seed release upon environmental conditions suitable for wind dispersal. In this context, we propose an interplay of humidity and temperature to be the external "drivers" for the initial cone opening, in which resin works as a crucial chemical-mechanical latch system., Competing Interests: The authors declare that this study received funding from BASF SE, Ludwigshafen, Germany. The funder was not involved in the study design, collection, analysis, interpretation of data, the writing of this article, or the decision to submit it for publication., (Copyright © 2022 Horstmann, Buchheit, Speck and Poppinga.)

The freshwater crustacean Daphnia is well known for its expression of morphological defenses in the presence of predators. Research into this phenomenon has mostly centered on the ecology and evolution of Daphnia defenses; information is limited on the cellular mechanisms that underlie site-specific tissue growth. We aimed to determine these cellular mechanisms, specifically those associated with the development of defensive crests in D. longicephala. With the help of a cell-proliferation assay we monitored changes in the epidermal tissue of naïve and predator-exposed D. longicephala. Based on our results, we propose that cell division is delayed in favor of cell growth, which results in crest formation. Further, we identify specific regions of proliferative activity in a time-dependent manner. Defense development starts in the ventral region, before extending in the cranial and then dorsal directions. We demonstrate that these cellular changes begin as early as 2 h after predator exposure. Our results provide new insights into the cellular processes underlying morphological defense expression in Daphnia ., Competing Interests: The authors declare no conflict of interest., (© 2022 The Author(s).)

Eusociality has evolved multiple times across the insect phylogeny. Social insects with greater levels of social complexity tend to exhibit specialized castes with low levels of individual phenotypic plasticity. In contrast, species with simple social groups may consist of totipotent individuals that transition among behavioral and reproductive states. However, recent work has shown that in simple social groups, there can still be constraint on individual plasticity, caused by differences in maternal nourishment or social interaction. It is not well understood how these constraints arise, ultimately leading to the evolution of nonreproductive workers. Some species of orchid bees form social groups of a dominant and-one to two subordinate helpers where all individuals are reproductive. Females can also disperse to start their own nest as a solitary foundress, which includes a nonreproductive phase characterized by ovary inactivation, not typically expressed by subordinates. Little is known about individual flexibility across these trajectories. Here, using the orchid bee Euglossa dilemma, we assess the plasticity of subordinate helpers, finding that they are capable of the same behavioral, physiological, transcriptomic, and chemical changes seen in foundresses. Our results suggest that the lack of nonreproductive workers in E. dilemma is not due to a lack of subordinate plasticity., (© 2022 The Authors. Evolution © 2022 The Society for the Study of Evolution.)

Background and Aims: Pollen limitation is most prevalent among bee-pollinated plants, self-incompatible plants and tropical plants. However, we have very little understanding of the extent to which pollen limitation affects fruit set in mass-flowering trees despite tree crops accounting for at least 600 million tons of the 9200 million tons of annual global food production., Methods: We determined the extent of pollen limitation in a bee-pollinated, partially self-incompatible, subtropical tree by hand cross-pollinating the majority of flowers on mass-flowering macadamia (Macadamia integrifolia) trees that produce about 200 000-400 000 flowers. We measured tree yield and kernel quality and estimated final fruit set. We genotyped individual kernels by MassARRAY to determine levels of outcrossing in orchards and assess paternity effects on nut quality., Key Results: Macadamia trees were pollen-limited. Supplementary cross-pollination increased nut-in-shell yield, kernel yield and fruit set by as much as 97, 109 and 92 %, respectively. The extent of pollen limitation depended upon the proximity of experimental trees to trees of another cultivar because macadamia trees were highly outcrossing. Between 84 and 100 % of fruit arose from cross-pollination, even at 200 m (25 rows) from orchard blocks of another cultivar. Large variations in nut-in-shell mass, kernel mass, kernel recovery and kernel oil concentration were related to differences in fruit paternity, including between self-pollinated and cross-pollinated fruit, thus demonstrating pollen-parent effects on fruit quality (i.e. xenia)., Conclusions: This study is the first to demonstrate pollen limitation in a mass-flowering tree. Improved pollination led to increased kernel yield of 0.31-0.59 tons ha-1, which equates currently to higher farm-gate income of approximately $US3720-$US7080 ha-1. The heavy reliance of macadamia flowers on cross-pollination and the strong xenia effects on kernel mass demonstrate the high value that pollination services can provide to food production., (© The Author(s) 2021. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Cross-pollination can improve fruit yield, fruit size and nutritional quality of many food crops. However, we rarely understand what proportions of the crop result from self- or cross-pollination, how cross-pollination affects crop quality, and how far pollen is transported by pollinators. Management strategies to improve pollination services are consequently not optimal for many crops. We utilised a series of SNP markers, unique for each cultivar of avocado, to quantify proportions of self- and cross-paternity in fruit of Hass avocado at increasing distances from cross-pollen sources. We assessed whether distance from a cross-pollen source determined the proportions of self-pollinated and cross-pollinated fruit, and evaluated how self- and cross-paternity affected fruit size and nutritional quality. Avocado fruit production resulted from both self- and cross-pollination in cultivar Hass in Queensland, Australia. Cross-pollination levels decreased with increasing distance from a cross-pollen source, from 63% in the row adjacent to another cultivar to 25% in the middle of a single-cultivar block, suggesting that pollen transport was limited across orchard rows. Limited pollen transport did not affect fruit size or quality in Hass avocados as xenia effects of a Shepard polliniser on size and nutritional quality were minor., (© 2021. The Author(s).)

Predation is a major selective agent, so that many taxa evolved phenotypically plastic defensive mechanisms. Among them are many species of the microcrustacean genus Daphnia, which respond to an increased predation risk by developing inducible morphological alterations. Some of these features are obvious and easily recognized, e.g., crests in D. longicephala, while others are rather hidden, such as the bulkier shape of D. magna induced by the presence of the tadpole shrimp Triops. In this study we investigated the extraordinary diversity of morphological adaptations in the presence of predators with different foraging strategies in six predator-prey systems. For the first time we were able to analyze the unexposed and predator-exposed morphs comprehensively using three-dimensional scanning and reconstruction. We show that morphological changes are manifold in appearance between species and predators, and go beyond what has been known from previous 2D analyses. This further demonstrates the enormous trait flexibility of Daphnia. Interestingly, we found that among this variety some species share morphological strategies to counter a predator, while others use a different strategy against the same predator. Based on these intra- and interspecific comparisons, we discuss the mechanisms by which the respective defense might operate. These data therefore contribute to a deeper understanding of the inducible defenses' morphology as well as their diversified modes of operation in Daphnia, being a cornerstone for subsequent investigations, including the determination of costs associated with morphological change., Competing Interests: The authors have declared that no competing interests exist.

Dormancy is a lifecycle delay that allows organisms to escape suboptimal environmental conditions. As a genetically programmed type of dormancy, diapause is usually accompanied by metabolic depression and enhanced tolerance toward adverse environmental factors. However, the drivers and regulators that steer an organism's development into a state of suspended animation to survive environmental stress have not been fully uncovered. Heat shock proteins 70 (HSP70s), which are often produced in response to various types of stress, have been suggested to play a role in diapause. Considering the diversity of the Hsp70 family, different family members may have different functions during diapause. In the present study, we demonstrate the expression of two hsp70 genes (A and B together with protein localization of B) throughout continuous and diapause interrupted development of Daphnia magna . Before and after diapause, the expression of Dmhsp70-A is low. Only shortly before diapause and during diapause, Dmhsp70-A is significantly upregulated and may therefore be involved in diapause preparation and maintenance. In contrast, Dmhsp70-B is expressed only in developing embryos but not in diapausing embryos. During continuous development, the protein of this Hsp70 family member is localized in the cytosol. When we expose both embryo types to heat stress, expression of both hsp70 genes increases only in developing embryos, and the protein of family member B is translocated to the nucleus. In this stress formation, this protein provides effective protection of nucleoplasmic DNA. As we also see this localization in diapausing embryos, it seems that Daphnia embryo types share a common subcellular strategy when facing dormancy or heat shock, i.e., they protect their DNA by HSP70B nuclear translocation. Our study underlines the distinctive roles that different Hsp70 family members play throughout continuous and diapause interrupted development., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Chen, Gómez and Weiss.)

Fossil fuel combustion results in rising atmospheric carbon dioxide (CO 2 ), which is known to impact the global climate and the oceans. Latest insights indicate that rising atmospheric CO 2 levels also affect CO 2 partial pressure ( p CO 2 ) in freshwaters, where p CO 2 is controlled by a multitude of parameters. However, up to date there is no standardized method, which allows the determination of current and past freshwater p CO 2 levels. Ideally methods should incorporate numerous hydrogeochemical and -physical factors to reflect the interplay of all interacting components and their effect on p CO 2 . We here describe the application of the geochemical program PHREEQC. This freeware serves as an easy method enabling a plausible and comprehensive analysis of p CO 2 for field, laboratory, and especially long-term data. We present the use of the different input parameters of a laboratory- and a field long-term monitoring dataset including dissociation constants of carbonic acid measured as total inorganic carbon (TIC) and total CO 2 concentration (TCO 2 ) or total alkalinity (TA), together with hydrogeochemical and -physical parameters. Based on current literature and our analyses PHREEQC appears a solid strategy to determine freshwater p CO 2 that can moreover be used for long-term datasets.•Comprehensive analysis of p CO 2 for field, laboratory, and long-term data.•PHREEQC is not dependent on just one sampling method or parameter scheme.•PHREEQC includes testing the plausibility of a water analysis and enables the assessment of the quality of the laboratory analysis, as well as automatic calculation of all relevant aquatic complexes., Competing Interests: 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., (© 2021 The Authors. Published by Elsevier B.V.)

Predator-induced phenotypic plasticity describes the ability of prey to respond to an increased predation risk by developing adaptive phenotypes. Upon the perception of chemical predator cues, the freshwater crustacean Daphnia longicephala develops defensive crests against its predator Notonecta spec. (Heteroptera). Chemical predator perception initiates a cascade of biological reactions that leads to the development of these morphological features. Neuronal signaling is a central component in this series, however how the nervous system perceives and integrates environmental signals is not well understood. As neuronal activity is often accompanied by functional and structural plasticity of the nervous system, we hypothesized that predator perception is associated with structural and functional changes of nervous tissues. We observe structural plasticity as a volume increase of the central brain, which is independent of the total number of brain cells. In addition, we find functional plasticity in form of an increased number of inhibitory post-synaptic sites during the initial stage of defense development. Our results indicate a structural rewiring of nerve-cell connections upon predator perception and provide important insights into how the nervous system of prey species interprets predator cues and develops cost-benefit optimized defenses.

Corals have evolved a variety of stress responses to changing conditions, many of which have been the subject of scientific research. However, polyp bailout has not received widespread scientific attention, despite being described more than 80 years ago. Polyp bailout is a drastic response to acute stress in which coral colonies break down, with individual and patches of polyps detaching from the colony and the calcareous skeleton Polyps retain their symbiotic partners, have dispersal ability, and may undergo secondary settlement and calcification. Polyp bailout has been described worldwide in a variety of anthozoan species, especially in Scleractinia. It can be induced by multiple natural stressors, but also artificially. Little is known about the evolutionary and ecological potential and consequences of breaking down modularity, the dispersal ability, and reattachment of polyps resulting from polyp bailout. It has been shown that polyp bailout can be used as a model system, with promise for implementation in various research topics. To date, there has been no compilation of knowledge on polyp bailout, which prompted us to review this interesting stress response and provide a basis to discuss research topics and priorities for the future., Competing Interests: None declared., (© 2021 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Background: Museums and other institutions curating natural history collections (NHCs) are fundamental entities to many scientific disciplines, as they house data and reference material for varied research projects. As such, biological specimens preserved in NHCs represent accessible physical records of the living world's history. They provide useful information regarding the presence and distribution of different taxonomic groups through space and time. Despite the importance of biological museum specimens, their potential to answer scientific questions, pertinent to the necessities of our current historical context, is often under-explored.The currently-known wild bee fauna of Luxembourg comprises 341 registered species distributed amongst 38 different genera. However, specimens stored in the archives of local NHCs represent an untapped resource to update taxonomic lists, including potentially overlooked findings relevant to the development of national conservation strategies., New Information: We re-investigated the wild bee collection of the Zoology Department of the National Museum of Natural History Luxembourg by using morphotaxonomy and DNA barcoding. The collection revision led to the discovery of four species so far not described for the country: Andrena lagopus (Latreille, 1809), Nomada furva (Panzer, 1798), Hoplitis papaveris (Latreille, 1799) and Sphecodes majalis (Pérez, 1903). Additionally, the presence of Nomada sexfasciata (Panzer, 1799), which inexplicably had been omitted by the most current species list, can be re-confirmed. Altogether, our findings increase the number of recorded wild bee species in Luxembourg to 346. Moreover, the results highlight the crucial role of NHCs as repositories of our knowledge of the natural world., (Fernanda Herrera Mesías, Alexander M. Weigand.)

Utricularia multifida is carnivorous bladderwort from Western Australia and belongs to a phylogenetically early-diverging lineage of the genus. We present a prey spectrum analysis resulting from a snapshot sampling of 17 traps-the first of this species to our knowledge. The most abundant prey groups were Ostracoda, Copepoda, and Cladocera. The genus cf. Cypretta (Cyprididae, Ostracoda) was the predominant prey. However, a high variety of other prey organisms with different taxonomic backgrounds was also detected. Our results indicate that U. multifida may potentially be specialized in capturing substrate-bound prey. Future approaches should sample plants from different localities to allow for robust comparative analyses., Competing Interests: The authors have declared that no competing interests exist.

Diapause is a mechanism necessary for survival in arthropods. Often diapause induction and resurrection is light-dependent and therefore dependent on the photoperiod length and on the number of consecutive short-days. In many organisms, including the microcrustacean Daphnia magna, one functional entity with the capacity to measure seasonal changes in day-length is the circadian clock. There is a long-standing discussion that the circadian clock also controls photoperiod-induced diapause. We tested this hypothesis in D. magna, an organism which goes into a state of suspended animation with the shortening of the photoperiod. We measured gene expression of clock genes in diapause-destined embryos of D. magna in the initiation, resting and resurrection phases and checked it against gene expression levels of continuously developing embryos. We demonstrate that some genes of the clock are differentially expressed during diapause induction but not during its maintenance. Furthermore, the photoreceptor gene cry2 and the clock-associated gene brp are highly expressed during induction and early diapause, probably in order to produce excess mRNA to prepare for immediate resurrection. After resurrection, both types of embryos show a similar pattern of gene expression during development. Our study contributes significantly to the understanding of the molecular basis of diapause induction, maintenance and termination.

Limited data pertaining to life history and population connectivity of the data-deficient southern stingray (Hypanus americanus) are available. To determine potential vulnerabilities of their populations, this study aimed to analyse their movement patterns and genetic variability. A population of southern stingrays encompassing nine sites around Cape Eleuthera, the Bahamas, has been monitored using mark-recapture, spanning a 2.5 year period. Out of 200 individual stingrays, more than a third were encountered again. The home range of the females appears to be restricted, which supports the notion of high site residency. As resident populations of stingrays could suffer from a lack of population connectivity and be predestined for genetic isolation and local extirpation, this study further investigated the genetic connectivity of four sample sites in the central and western Bahamas. A haplotype analysis from the mitochondrial D-loop region showed that no distinct population structure strictly correlated with the sample site. These findings were complemented by five microsatellite loci that revealed high degrees in genotypic variability and little population differentiation. The results suggest gene flow mediated by both males and females., (© 2020 The Authors. Journal of Fish Biology published by John Wiley & Sons Ltd on behalf of The Fisheries Society of the British Isles.)

Tree nuts play an important role in healthy diets, but their economic value and nutritional quality may be affected by their size and paternity. We assessed relationships between nut size and kernel recovery, the incidence of whole kernels, fatty acid composition and mineral nutrient concentrations in three macadamia cultivars, "Daddow", "816" and "A4". We determined to what extent differences in nut size and quality were the result of different levels of cross- or self-paternity. Small nuts of all cultivars had lower kernel recovery than large nuts, and small nuts provided lower incidence of whole kernels in "Daddow" and "A4". Small kernels had a lower relative abundance of the saturated fatty acid, palmitic acid, in all cultivars and higher relative abundance of the unsaturated fatty acid, oleic acid, in "Daddow" and "A4". Small kernels had higher concentrations of many essential nutrients such as nitrogen and calcium, although potassium concentrations were lower in small kernels. Most nuts arose from cross-pollination. Therefore, nut size and kernel quality were not related to different levels of cross- and self-paternity. Identified cross-paternity was 88%, 78% and 90%, and identified self-paternity was 3%, 2% and 0%, for "Daddow", "816" and "A4", respectively. Small macadamia kernels are at least as nutritious as large macadamia kernels. High levels of cross-paternity confirmed that many macadamia cultivars are predominantly outcrossing. Macadamia growers may need to closely inter-plant cultivars and manage beehives to maximise cross-pollination., Competing Interests: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Aquatic acidification is a major consequence of fossil fuel combustion. In marine ecosystems it was shown, that increasing pCO 2 levels significantly affect behavioural and sensory capacities in a diversity of species. This can result in altered predator and prey interactions and thereby change community structures. Just recently also CO 2 dependent acidification of freshwater habitats has been shown. Also here, increased levels of pCO 2 change organisms' behaviour and sensory capacities. For example, the freshwater crustacean Daphnia's ability to detect predators and accurately develop morphological defences was significantly reduced, rendering Daphnia more susceptible to predation. It was speculated that this may have cascading effects on freshwater food webs. However, for a comprehensive understanding of how increased levels of CO 2 affect trophic interactions, it is also important to study how CO 2 affects predators. We tested this using the dipeteran phantom midge larva Chaoborus obscuripes, which is a world-wide abundant inhabitant of freshwater impoundments. We monitored activity parameters, predation parameters, and predation rate. Chaoborus larvae are affected by increased levels of pCO 2 as we observed an increase in undirected movements and at the same time, reduced sensory abilities to detect prey items. This is likely to affect the larvae's energy budgets. Chaoborus is a central component of many freshwater food-webs. Therefore, CO 2 effects on predator and prey levels will likely have consequences for community structures.

Sexual signaling is an important reproductive barrier known to evolve early during the formation of new species, but the genetic mechanisms that facilitate the divergence of sexual signals remain elusive. Here we isolate a gene linked to the rapid evolution of a signaling trait in a pair of nascent neotropical orchid bee lineages, Euglossa dilemma and E. viridissima. Male orchid bees acquire chemical compounds from their environment to concoct species-specific perfumes to later expose during courtship. We find that the two lineages acquire chemically distinct perfumes and are reproductively isolated despite low levels of genome-wide differentiation. Remarkably, variation in perfume chemistry coincides with rapid divergence in few odorant receptor (OR) genes. Using functional assays, we demonstrate that the derived variant of Or41 in E. dilemma is specific towards its species-specific major perfume compound, whereas the ancestral variant in E. viridissima is broadly tuned to multiple odorants. Our results show that OR evolution likely played a role in the divergence of sexual communication in natural populations.

We investigated the predator-prey interactions between an Australian ecotype of the carnivorous waterwheel plant (Aldrovanda vesiculosa, Droseraceae) and its potential natural prey, the water flea Daphnia longicephala (Daphniidae), which also occurs in Australia. A. vesiculosa develops snap-traps, which close within ~10-100 ms after mechanical triggering by zooplankton prey. Prey capture attempts (PCAs) were recorded via high-speed cinematography in the laboratory. From 14 recorded PCAs, nine were successful for the plant (the prey was caught), and five were unsuccessful (prey could escape), resulting in a capture rate of ~64%. The prey animals' locomotion behaviour (antenna beat frequency and movement type) in trap vicinity or inside the open traps is very variable. Traps were mainly triggered with the second antennae. During trap closure, the animals moved only very little actively. A flight response in reaction to an initiated trap closure was not observed. However, several animals could escape, either by having a "lucky" starting position already outside the triggered trap, by freeing themselves after trap closure, or by being pressed out by the closing trap lobes. According to our observations in the successful PCAs, we hypothesize that the convex curvature of the two trap lobes (as seen from the outside) and the infolded trap rims are structural means supporting the capture and retention of prey. Our results are discussed in a broader biological context and promising aspects for future studies are proposed.

In accordance with the Red Queen hypothesis, the lower genotypic diversity in clonally reproducing species should make them easier targets for pathogen infection, especially when closely related sexually reproducing species occur in close proximity. We analyzed two populations of clonal P. formosa and their sexual parental species P. mexicana by correlating individual parasite infection with overall and immune genotype. Our study revealed lower levels of overall genotypic diversity and marginally fewer MHC class I alleles in P. formosa individuals compared to sexually reproducing P. mexicana. Parasite load, however, differed only between field sites but not between species. We hypothesize that this might be due to slightly higher genotypic diversity in P. formosa at the innate immune system (toll like receptor 8) which is likely due to the species' hybrid origin. In consequence, it appears that clonal individuals do not necessarily suffer a disadvantage compared to sexual individuals when fighting parasite infection., Competing Interests: The authors have declared that no competing interests exist.

Male orchid bees (Euglossini) collect volatiles from their environment to concoct species-specific "perfumes", which are later emitted at mating sites. Intensity, complexity or composition of perfumes may encode age (survival) of a male, but how the individual perfume develops over time needs to be clarified. We investigated chemical changes during storage in leg pockets. We injected a mixture of eight perfume compounds into pockets of Euglossa imperialis and only the two most volatile compounds decreased over 12 days. Using a different approach we found significant shifts in quantities of naturally occurring perfume compounds of Euglossa championi over 10 days, with the strongest decreases (up to 70% peak area) in highly volatile minor compounds, e.g. monoterpenes, and noteworthy increases (up to 40%) in some sesquiterpenoids. Corresponding shifts were observed in legs of dried bees, suggesting that no metabolic activity is required for the observed changes to occur. Our results confirm that male orchid bees are generally good at preserving collected perfumes. However, subtle shifts towards heavier compounds in blends may occur over the lifetime of individual bees, e.g. due to evaporation or in-pocket chemical reaction, with old males acquiring a more pronounced base note in their seasoned perfumes.

The critically endangered carnivorous waterwheel plant ( Aldrovanda vesiculosa , Droseraceae) possesses underwater snap traps for capturing small aquatic animals, but knowledge on the exact prey species is limited. Such information would be essential for continuing ecological research, drawing conclusions regarding trapping efficiency and trap evolution, and eventually, for conservation. Therefore, we performed comparative trap size measurements and snapshot prey analyses at seven Czech and one German naturalized microsites on plants originating from at least two different populations. One Czech site was sampled twice during 2017. We recorded seven main prey taxonomic groups, that is, Cladocera, Copepoda, Ostracoda, Ephemeroptera, Nematocera, Hydrachnidia, and Pulmonata. In total, we recorded 43 different prey taxa in 445 prey-filled traps, containing in sum 461 prey items. With one exception, prey spectra did not correlate with site conditions (e.g. water depth) or trap size. Our data indicate that A. vesiculosa shows no prey specificity but catches opportunistically, independent of prey species, prey mobility mode (swimming or substrate-bound), and speed of movement. Even in cases where the prey size exceeded trap size, successful capture was accomplished by clamping the animal between the traps' lobes. As we found a wide prey range that was attracted, it appears unlikely that the capture is enhanced by specialized chemical- or mimicry-based attraction mechanisms. However, for animals seeking shelter, a place to rest, or a substrate to graze on, A. vesiculosa may indirectly attract prey organisms in the vicinity, whereas other prey capture events (like that of comparably large notonectids) may also be purely coincidental., (© The Author(s) 2019. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology.)

Filter feeding zooplankton are a crucial component of limnic food webs. Copepods and cladocerans are important prey organisms for first-level predators like the common and abundant larvae of phantom midges (Chaoborus sp.). The latter possess a complex catching basket built of head appendages specialized to capture small crustaceans. The predator-prey-relationship of Chaoborus (Diptera, Nematocera) and Daphnia (Crustacea, Cladocera) has been studied in particular detail owing to the daphniids' ability to react upon the threat of predation with inducible defenses. Daphnia pulex expresses so-called 'neckteeth' in the presence of Chaoborus larvae that are discussed as a defensive trait that interferes with the larval head appendages and their effectiveness has been shown in several studies. Nonetheless, mode of function of these neckteeth is not understood and the hypothesis that they interfere with the predator's head appendages still has to be confirmed. To clarify the role of neckteeth in Daphnia, an understanding of the Chaoborus capture apparatus is essential. Here, we present a detailed three-dimensional analysis of Chaoborus obscuripes' larval head morphology as well as a kinematic analysis of the attack motion, which revealed an impressive strike velocity (14 ms to prey contact). The movement of the larvae's head appendages is reconstructed in the three-dimensional space using a combination of high-speed videography, micro-computed tomography and computer animation. Furthermore, we provide predation trial data to distinguish between pre- and post-attack defensive effects in D. pulex. Our findings suggest a combination of pre- and post-attack defenses with an average effectiveness of 50% each. With this study, we quantitatively describe prey capture kinematics of C. obscuripes and take a further step to reveal the neckteeth' mode of function in D. pulex., Competing Interests: The authors have declared that no competing interests exist.

Ecological communities are organized in trophic levels that share manifold interactions forming complex food webs. Infochemicals can further modify these interactions, e.g., by inducing defenses in prey. The micro-crustacean Daphnia is able to respond to predator-specific chemical cues indicating an increased predation risk. Daphnia shows plastic responses by adapting its morphology, behavior, and physiology, increasing organism, and population fitness. This stabilizes community structures. This review will describe the progress that has been made in understanding the high degree of plasticity observed in the model crustacean Daphnia . I summarize current knowledge on the processes of predator detection, ranging from the nature of biologically active chemical cues to the underlying neurophysiological mechanisms. With this, I aim to provide a comprehensive overview on the molecular mechanisms of ad hoc environmental phenotypic adaptation. In times of climate change and pollution understanding information transfer in aquatic systems is valuable as it will allow us to predict whether and how community structures are being affected.

Background: Diapause is a form of dormancy that is genetically predetermined to allow animals to overcome harsh environmental conditions. It is induced by predictive environmental cues bringing cellular activity levels into a state of suspended animation. Entering diapause requires organismal, molecular and cellular adaptation to severely reduced energy flows. Cells must therefore have evolved strategies that prepare them for periods with limited metabolic resources. However, changes that occur on the (sub-)cellular level have not been thoroughly described., Results: We investigated mitotic activity and we monitored cytoskeletal network changes in successive stages of diapausing and non-diapausing Daphnia magna embryos using (immuno-)fluorescent labeling. We find that embryos destined to diapause show a delayed and 2.5x slower mitotic activity in comparison to continuously developing embryos. Development is halted when D. magna embryos reach ~ 3500 cells, whereupon mitotic activity is absent and cytoskeletal components are severely reduced, rendering diapause cells compact and condensed., Conclusion: In the initiation phase of diapause, the slower cell division rate points to prolonged interphase duration, preparing the cells for diapause maintenance. During diapause, cytoskeletal depletion and cellular condensation may be a means to save energy resources. Our data provide insights into the sub-cellular change of diapause in Daphnia.

After infection with Trypanosoma cruzi , the etiologic agent of Chagas disease, immunosuppression, and apoptosis of mature lymphocytes contribute to the establishment of the parasite in the host and thereby to persistence and pathology in the chronic stage of infection. In a systemic mouse model of experimental Chagas disease, we have demonstrated a strong depletion of mature B cells in the spleen during the first 2 weeks of infection. Remarkably, the decrease in this cell population commenced already in the bone marrow from infected mice and was a concomitant of an increased apoptosis in pro- and pre-B cell populations. Pro- and pre-B cells in the bone marrow showed a significant reduction accompanied by a functional disturbance of bone marrow-derived stromal cells resulting in diminished levels of IL-7, an essential factor for the development of B cell precursors. Ex vivo , stromal cells isolated from the bone marrow of infected mice had a strikingly impaired capacity to maintain the development of pro- and pre-B cells obtained from uninfected animals. Together, the reduction of an active humoral immune response during acute Chagas disease suggests to be an initial immune evasion mechanism of the parasite to establish persistent infection. Therefore, prevention of B cell depletion by rescuing the stromal cells during this early phase, could give rise to new therapeutic approaches.

Infochemicals play important roles in aquatic ecosystems. They even modify food web interactions, such as by inducing defenses in prey. In one classic but still not fully understood example, the planktonic freshwater crustacean Daphnia pulex forms specific morphological defenses (neckteeth) induced by chemical cues (kairomones) released from its predator, the phantom midge larva Chaoborus. On the basis of liquid chromatography, mass spectrometry, and chemical synthesis, we report here the chemical identity of the Chaoborus kairomone. The biologically active cues consist of fatty acids conjugated to the amino group of glutamine via the N terminus. These cues are involved in Chaoborus digestive processes, which explains why they are consistently released despite the disadvantage for its emitter. The identification of the kairomone may allow in-depth studies on multiple aspects of this inducible defense system.

Quantitative analysis of shape and form is critical in many biological disciplines, as context-dependent morphotypes reflect changes in gene expression and physiology, e.g., in comparisons of environment-dependent phenotypes, forward/reverse genetic assays or shape development during ontogenesis. 3D-shape rendering methods produce models with arbitrarily numbered, and therefore non-comparable, mesh points. However, this prevents direct comparisons. We introduce a workflow that allows the generation of comparable 3D models based on several specimens. Translocations between points of modelled morphotypes are plotted as heat maps and statistically tested. With this workflow, we are able to detect, model and investigate the significance of shape and form alterations in all spatial dimensions, demonstrated with different morphotypes of the pond-dwelling microcrustacean Daphnia . Furthermore, it allows the detection even of inconspicuous morphological features that can be exported to programs for subsequent analysis, e.g., streamline- or finite-element analysis., Competing Interests: The authors declare there are no competing interests.

The extreme rarity of asexual vertebrates in nature is generally explained by genomic decay due to absence of meiotic recombination, thus leading to extinction of such lineages. We explore features of a vertebrate asexual genome, the Amazon molly, Poecilia formosa, and find few signs of genetic degeneration but unique genetic variability and ongoing evolution. We uncovered a substantial clonal polymorphism and, as a conserved feature from its interspecific hybrid origin, a 10-fold higher heterozygosity than in the sexual parental species. These characteristics seem to be a principal reason for the unpredicted fitness of this asexual vertebrate. Our data suggest that asexual vertebrate lineages are scarce not because they are at a disadvantage, but because the genomic combinations required to bypass meiosis and to make up a functioning hybrid genome are rarely met in nature.

The activity patterns of trunk muscles are commonly neglected, in spite of their importance for maintaining body shape. Analysis of the biomechanics of the trunk under static conditions has led to predictions of the activity patterns. These hypotheses are tested experimentally by surface electromyography (EMG). Five horses, with and without a rider, were examined in the walk, trot and canter. Footfall was synchronised with EMG by an accelerometer. Averages of ten consecutive cycles were calculated and compared by statistical methods. The start and stop times of the muscle activities of 5-10 undisturbed EMG plots were determined and the averages and standard deviations calculated. In walking, muscle activities are minor. Electromyography (EMG) activity was increased in the m. rectus during the three-limb support. When the bending moments assume their greatest values, for example while the horses' mass is accelerated upward (two times earth acceleration) in the diagonal support phases in trot and canter the m. rectus, connecting the sternum with the pubic bone is most active. The m. obl. externus is most active when the torsional and bending moments are greatest during the same support phases, but not bilaterally, because the forces exerted on one side by the (recorded) m. obl. externus are transmitted on the other side by the (not recorded) m. obl. internus. While the hindlegs touch the ground in the trot and canter, ground reaction forces tend to flex the hip joint and the lumbar spine. Therefore, the vertebral column needs to be stabilised by the ipsilateral m. longissimus dorsi, which in fact can be observed. As a whole, our EMG data confirm exactly what has been predicted by theoretical analysis., (© 2017 Blackwell Verlag GmbH.)

Stream ecosystems are impacted by multiple stressors worldwide. Recent studies have shown that the effects of multiple stressors are often complex and difficult to predict based on the effects of single stressors. More research is needed to understand stressor impacts on stream communities and to design appropriate counteractions. We carried out an outdoor mesocosm experiment to assess single and interactive multiple-stressor effects on stream macroinvertebrates in a setup with controlled application of three globally important stressors, namely, reduced stream flow velocity, deposition of fine sediment and increased chloride concentration in a full-factorial design. Each mesocosm comprised three compartments (channel substratum, leaf litter bag and drift net) that were individually analyzed and also compared. We identified 102,501 specimens in total (mainly to family level), 36.5% of which were found in the substratum, 60.6% in litter bags and 2.9% in the drift. Added fine sediment and reduced flow velocity had strong negative single-stressor effects on the abundances of EPT taxa, i.e. Ephemeroptera (mayflies), Plecoptera (stoneflies) and Trichoptera (caddisflies), and a positive effect on chironomid abundances in the substratum. Increased salt concentration reduced abundances of Ephemeroptera. Chironomids migrated from litter bag to channel substratum when water velocity was reduced and Leptophlebiidae in the opposite direction when sediment was added. All three stressors caused higher drift propensities, especially added fine sediment. Both additive and complex multiple-stressor effects were common. A complex three-way interaction affected EPT richness in the substratum, demonstrating the need to evaluate higher-order interactions for more than two stressors. Our results add further evidence that multiple-stressor interactions, notably increased salinity with other stressors, affect a variety of invertebrate taxa across different habitats of stream communities. The results have direct implications for water management as they highlight the need to re-evaluate defined salinity thresholds in the context of multiple-stressor interactions., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: Scavenger guilds are composed of a variety of species, co-existing in the same habitat and sharing the same niche in the food web. Niche partitioning among them can manifest in different feeding strategies, e.g. during carcass feeding. In the bentho-pelagic realm of the Southern Ocean, scavenging amphipods (Lysianassoidea) are ubiquitous and occupy a central role in decomposition processes. Here we address the question whether scavenging lysianassoid amphipods employ different feeding strategies during carcass feeding, and whether synergistic feeding activities may influence carcass decomposition. To this end, we compared the relatively large species Waldeckia obesa with the small species Cheirimedon femoratus, Hippomedon kergueleni, and Orchomenella rotundifrons during fish carcass feeding ( Notothenia spp.). The experimental approach combined ex situ feeding experiments, behavioural observations, and scanning electron microscopic analyses of mandibles. Furthermore, we aimed to detect ecological drivers for distribution patterns of scavenging amphipods in the Antarctic coastal ecosystems of Potter Cove. In Potter Cove, the climate-driven rapid retreat of the Fourcade Glacier is causing various environmental changes including the provision of new marine habitats to colonise. While in the newly ice-free areas fish are rare, macroalgae have already colonised hard substrates. Assuming that a temporal dietary switch may increase the colonisation success of the most abundant lysianassoids C. femoratus and H. kergueleni , we aimed to determine their consumption rates (g food x g amphipods -1 x day -1 ) and preferences of macroalgae and fish., Results: We detected two functional groups with different feeding strategies among scavenging amphipods during carcass feeding: carcass 'opener' and 'squeezer'. Synergistic effects between these groups were not statistically verified under the conditions tested. C. femoratus switched its diet when fish was not available by consuming macroalgae (about 0.2 day -1 ) but preferred fish by feeding up to 80% of its own mass daily. Contrary, H. kergueleni rejected macroalgae entirely and consumed fish with a maximal rate of 0.8 day -1 ., Conclusion: This study reveals functional groups in scavenging shallow-water amphipods and provides new information on coastal intraguild niche partitioning. We conclude that the dietary flexibility of C. femoratus is a potential ecological driver and central to its success in the colonisation of newly available ice-free Antarctic coastal habitats., Competing Interests: In the framework of the research project IMCONet, PICTO 0116–2012-2016 and PICTO 0100–2010 the Environmental and Tourism Antarctic Management Program of the National Direction of the Antarctic (Dirección Nacional del Antártico) in the Republic of Argentina, has issued the appropriate permissions to all the stages of this research. This regards the Antarctic Specially Protected Area N°132 “Peninsula Potter” (under art. 7, Annex V of the Madrid Protocol, Law 25,260) and taking and harmful interference and introduction of species (under art. 3, Annex II of the Madrid Protocol, Law 24,216). Both of these permissions properly followed the regulations in force.Not applicable.The authors declare that they have no competing interest.Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

The freshwater crustacean Daphnia is known for its ability to develop inducible morphological defences that thwart predators. These defences are developed only in the presence of predators and are realized as morphological shape alterations e.g. 'neckteeth' in D. pulex and 'crests' in D. longicephala. Both are discussed to hamper capture, handling or consumption by interfering with the predator's prey capture devices. Additionally, D. pulex and some other daphniids were found to armour-up and develop structural alterations resulting in increased carapace stiffness. We used scanning transmission electron microscopy (STEM) and confocal laser scanning microscopy (CLSM) to identify predator-induced structural and shape alterations. We found species specific structural changes accompanying the known shape alterations. The cuticle becomes highly laminated (i.e. an increased number of layers) in both species during predator exposure. Using nano- and micro-indentation as well as finite element analysis (FEA) we determined both: the structure's and shape's contribution to the carapace's mechanical resistance. From our results we conclude that only structural alterations are responsible for increased carapace stiffness, whereas shape alterations appear to pose handling difficulties during prey capture. Therefore, these defences act independently at different stages during predation.

We recorded capture events (CEs) of the daphniid Ceriodaphnia dubia by the carnivorous Southern bladderwort with suction traps (Utricularia australis). Independent to orientation and behavior during trap triggering, the animals were successfully captured within 9 ms on average and sucked in with velocities of up to 4 m/s and accelerations of up to 2800 g. Phases of very high acceleration during onsets of suction were immediately followed by phases of similarly high deceleration (max.: -1900 g) inside the bladders, leading to immobilization of the prey which then dies. We found that traps perform a 'forward strike' during suction and that almost completely air-filled traps are still able to perform suction. The trigger hairs on the trapdoors can undergo strong bending deformation, which we interpret to be a safety feature to prevent fracture. Our results highlight the elaborate nature of the Utricularia suction traps which are functionally resilient and leave prey animals virtually no chance to escape.

Many insects rely on chemical signals to transmit precise information on the location, identity, and quality of potential mates. Chemical signals are often broadcasted at sites with physical properties that maximize signal propagation and signal transmission. Male neotropical orchid bees (Euglossini) perch and display on vertical branches and tree trunks in the forest to expose volatile blends (perfumes) that they previously collected from their environment. Previous studies have shown that the chemical composition of perfume blends is highly differentiated even between closely related species. However, variation in behavioral components of perfume exposure and male display remain poorly understood. We conducted a four-year study on orchid bee display sites (8 species) in pacific Costa Rica, using field observations along with chemical analysis and cage experiments to assess display niche partitioning among sympatric species. We evaluated the influence of physical factors (terrain, wind, light) on the distribution of perch sites and on display behavior, and tested a prediction of the sex pheromone-analogue hypothesis, i.e., that displaying males have above-average quantities or qualities of acquired perfumes. Males of different species displayed in the same general area and sometimes in close proximity to each other, but partitioned the display niche by selecting different perch diameters, display heights, and by displaying at different times of the day. Most perch sites were located inside the forest on elevated ground, especially along ridges, where stronger winds may help disperse perfume signals. Furthermore, the angular position of displaying males on perches was narrowly determined by wind direction, with males being positioned on the downwind side of the perch, where they would be most conspicuous to conspecifics approaching on an odor trail. Although our results generally support the hypothesis that perfumes serve as pheromone analogues, we did not find differences in the perfume composition of males caught at display perches and males captured at chemical baits. This suggests that, while chemical communication is an integral part of orchid bee display, male display activity is not determined by the history of, and success in, volatile acquisition., (© 2017 by the Ecological Society of America.)

Bioluminescence is a fascinating phenomenon occurring in numerous animal taxa in the ocean. The reef dwelling splitfin flashlight fish (Anomalops katoptron) can be found in large schools during moonless nights in the shallow water of coral reefs and in the open surrounding water. Anomalops katoptron produce striking blink patterns with symbiotic bacteria in their sub-ocular light organs. We examined the blink frequency in A. katoptron under various laboratory conditions. During the night A. katoptron swims in schools roughly parallel to their conspecifics and display high blink frequencies of approximately 90 blinks/minute with equal on and off times. However, when planktonic prey was detected in the experimental tank, the open time increased compared to open times in the absence of prey and the frequency decreased to 20% compared to blink frequency at night in the absence of planktonic prey. During the day when the school is in a cave in the reef tank the blink frequency decreases to approximately 9 blinks/minute with increasing off-times of the light organ. Surprisingly the non-luminescent A. katoptron with non-functional light organs displayed the same blink frequencies and light organ open/closed times during the night and day as their luminescent conspecifics. In the presence of plankton non-luminescent specimens showed no change in the blink frequency and open/closed times compared to luminescent A. katoptron. Our experiments performed in a coral reef tank show that A. katoptron use bioluminescent illumination to detect planktonic prey and that the blink frequency of A. katoptron light organs follow an exogenous control by the ambient light., Competing Interests: The authors have declared that no competing interests exist.

Theory predicts that both stabilising selection and diversifying selection jointly contribute to the evolution of sexual signalling traits by (1) maintaining the integrity of communication signals within species and (2) promoting the diversification of traits among lineages. However, for many important signalling traits, little is known about whether these dynamics translate into predictable macroevolutionary signatures. Here, we test for macroevolutionary patterns consistent with sexual signalling theory in the perfume signals of neotropical orchid bees, a group well studied for their chemical sexual communication. Our results revealed both high species-specificity and elevated rates of evolution in perfume signals compared to nonsignalling traits. Perfume complexity was correlated with the number of congeners in a species' range, suggesting that perfume evolution may be tied to the remarkably high number of orchid bee species coexisting together in some neotropical communities. Finally, sister-pair comparisons were consistent with both rapid divergence at speciation and character displacement upon secondary contact. Together, our results provide new insight into the macroevolution of sexual signalling in insects., (© 2016 The Authors. Ecology Letters published by CNRS and John Wiley & Sons Ltd.)

Background: Quantification of synapses and their morphological analysis are extensively used in network development and connectivity studies, drug screening and other areas of neuroscience. Thus, a number of quantitative approaches were introduced so far. However, most of the available methods are highly tailored to specific applications and have limitations for widespread use., New Method: We present a new plugin for the open-source software ImageJ to provide a modifiable, high-throughput and easy to use method for synaptic puncta analysis. Our approach is based on colocalization of pre- and postsynaptic protein markers. Structurally completed glutamatergic and GABAergic synapses were identified by VGLUT1-PSD95 and VGAT-gephyrin colocalization, respectively. By combining conventional confocal microscopy with stimulated emission depletion (STED) imaging, we propose a method to quantify the number of scaffolding protein clusters, recruited to a single postsynaptic density., Results: In a proof-of-concept study, we reveal the differential distribution of glutamatergic and GABAergic synapse density with reference to perineuronal net (PNN) expression. Using super-resolution STED imaging, we demonstrate that postsynaptic puncta of completed synapses are composed of significantly more protein clusters, compared to uncompleted synapses., Comparison With Existing Methods: Our Synapse Counter plugin for ImageJ offers a rapid and unbiased research tool for a broad spectrum of neuroscientists. The proposed method of synaptic protein clusters quantification exploits super-resolution imaging to provide a comprehensive approach to the analysis of postsynaptic density composition., Conclusions: Our results strongly substantiate the benefits of colocalization-based synapse detection., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Daphnia (Crustacea, Cladocera) are well known for their ability to form morphological adaptations to defend against predators. In addition to spines and helmets, the carapace itself is a protective structure encapsulating the main body, but not the head. It is formed by a double layer of the integument interconnected by small pillars and hemolymphatic space in between. A second function of the carapace is respiration, which is performed through its proximal integument. The interconnecting pillars were previously described as providing higher mechanical stability against compressive forces. Following this hypothesis, we analyzed the carapace structure of D. pulex using histochemistry in combination with light and electron microscopy. We found the distal integument of the carapace to be significantly thicker than the proximal. The pillars appear fibrous with slim waists and broad, sometimes branched bases where they meet the integument layers. The fibrous structure and the slim-waisted shape of the pillars indicate a high capacity for withstanding tensile rather than compressive forces. In conclusion they are more ligaments than pillars. Therefore, we measured the hemolymphatic gauge pressure in D. longicephala and indeed found the hemocoel to have a pressure above ambient. Our results offer a new mechanistic explanation of the high rigidity of the daphniid carapace, which is probably the result of a light-weight construction consisting of two integuments bound together by ligaments and inflated by a hydrostatic hyper-pressure in the hemocoel. J. Morphol. 277:1320-1328, 2016. © 2016 Wiley Periodicals, Inc., (© 2016 Wiley Periodicals, Inc.)

The mitochondrial genomes of the caddisfly species Sericostoma personatum and Thremma gallicum were sequenced on a 454 FLX and Illumina MiSeq platform, respectively. Reads were assembled de novo and remaining gaps in the S. personatum mitogenome closed by Sanger sequencing. The lengths of the assembled mitogenomes were 15,260 bp and 15,343 bp for S. personatum and T. gallicum, respectively. Both mitogenomes contained all 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, and the control region. The mitochondrial gene order of both caddisflies is identical with the typical insect gene order. These are the third and fourth published mitogenomes of the order Trichoptera of two formerly unexplored families and thus will be useful in future phylogenetic analysis.