Frontiers in Ecology and Evolution

Riparian environments are characterised by a high diversity of arthropod species, linking the aquatic with the terrestrial ecosystem. One of the dominant arthropod predators in this ecotone, carabid beetles are of particular interest as they may also act as facultative scavengers, feeding on carrion deposited along the shoreline. To test whether carabids feed on carrion, we examined the feeding preferences of the abundant riparian carabid Bembidion elongatum in a laboratory feeding trial, offering freshly killed and 24 hours post mortem Drosophila melanogaster. We subsequently assessed the detection probability of ribosomal prey RNA and DNA in predator gut contents using Drosophila-specific RT-PCR and PCR assays, and quantified nucleotide abundance by quantitative real-time PCR at 0, 3, 6, and 12 hours post-feeding. In the feeding experiment, B. elongatum showed a significant preference for fresh over carrion prey. Following consumption, the quantities of prey DNA and RNA in the predators gut declined over a 12-hour post-feeding period. However, no differences were detected in prey DNA or RNA quantities between individuals fed fresh prey and those fed carrion. Only immediately after consumption was the DNA:RNA ratio significantly lower in individuals fed fresh prey compared to those fed carrion while this difference was not observed at later time points. Overall, our results indicate that ingested ribosomal prey RNA in predators is present in high quantities, and that the DNA:RNA ratio is not a suitable indicator for distinguishing between consumption of carrion and fresh prey.

Environmental DNA (eDNA) offers a non-invasive alternative to traditional, more destructive sampling methods for determining species occupancy at ecological sites of interest. Aquatic eDNA sampling entails filtering a known volume of water to capture and detect genetic material shed by organisms. While the influence of individual environmental properties on the presence of target eDNA has been widely studied, it remains unclear how variables like temperature, pH, flow rate and conductivity correlate collectively with site electrofishing counts and eDNA concentrations. Resolving this question is important for two reasons: (1) typically, only eDNA, not physical specimens, is collected and measured, and (2) when methods like electrofishing and eDNA sampling are used in tandem, results often differ. Here unsupervised association rule-based machine learning is employed to discover interesting relationships among sampled covariates within a previously published case study of native brook trout (Salvelinus fontinalis) collected from Hanlon Creek (Guelph, Ontario, Canada) in September 2019. From a dataset of only 126 observations, the mining process revealed over 12000 plausible association rules linking covariates to eDNA concentrations (low/high) and electrofishing outcomes (absence/presence of brook trout). A strict pruning strategy reduced this ruleset to a manageable size of 153 associations, some of which were corroborated by existing literature, and some of which were novel (such as those potentially relating electrical conductivity to microbial and enzymatic activity). The entire workflow is included as a new R package called RulesTools. These results highlight the promise of association rule mining as a tool for guiding eDNA metadata collection, complementing statistical modelling, and informing conservation and management decision-making.

Many insect populations are experiencing thermal stress as a result of global change, making it imperative to investigate how their relationship with other organisms will be impacted by heat disturbances. Microbial symbionts, such as bacteria, have the potential to enhance or inhibit an insect's thermal tolerance. Solitary bee larvae host bacteria within their food stores ("pollen provisions"), which have been shown to benefit survival and development; however, it is unclear how heatwaves brought about by climate change will impact their relationships with these bacterial partners. In this study, we subjected blue orchard bee (Osmia lignaria) eggs and larvae to a 4-day heatwave (35 {degrees}C daytime:22 {degrees}C nighttime) or kept them at control temperatures (25 {degrees}C daytime:15 {degrees}C nighttime), then returned all bees to control temperatures for a 5-day recovery period. We assessed bacterial communities within pollen provisions and larval development stage pre-heatwave (Day 0), immediately post-heatwave (Day 4), and following the recovery period (Day 9). Bacterial community composition, diversity, and abundance were resilient to heat stress, but larval bees developed faster when subjected to a heatwave. This finding refutes the hypothesis that bacteria within pollen provisions modulate blue orchard bee responses to heat, suggesting instead that developmental effects could be more largely shaped by bee physiology or interactions with microorganisms other than bacteria.

A central goal when assessing patterns of population structure for conservation is to identify populations with unique genetic compositions. The use of genomic sequencing to identify distinct populations has become an increasingly popular method of delineating conservation units. Reduced costs associated with sequencing make it possible to generate larger, more informative datasets to assess genetic diversity within and among populations. In species that exhibit nest-site philopatry, genetic population structure can emerge on much finer scales, particularly in maternally inherited mitochondrial genomes. We demonstrate the feasibility and insight gained by using whole mitochondrial genome sequencing for evaluating population genetic structure and comparing to previous single marker studies in a vulnerable turtle. We used whole mitochondrial genome sequences from diamond-backed terrapin (Malaclemys terrapin) to evaluate whether nest-site philopatry generates fine-scale genetic structure among M. terrapin nesting beaches in western Mobile Bay (Alabama, USA). We then compared haplotype diversity between the Alabama population and M. terrapin populations from the Atlantic and Gulf coasts and evaluated the utility of using whole mitochondrial genomes rather than a subset of loci to characterize unique haplotypic diversity. We found no genetic structure associated with nest-site philopatry within Alabama, but none of the haplotypes in this region were shared with other Gulf Coast sites. This genetic structure is consistent with strong female natal philopatry within western Mobile Bay relative to the Gulf of Mexico and suggests that the Mobile Bay population is genetically unique relative to other M. terrapin populations and merits a unique conservation and management plan.

The charismatic migratory monarch butterfly population has declined dramatically, likely precipitated by loss of its breeding host plants (milkweed). Whether restoring milkweed would allow monarch recovery depends on whether additional factors currently limit the population. We investigated road mortality as one such factor. Monarchs cross thousands of roads during fall migration, and traffic volume has increased sharply while the population has plummeted. Using estimates of pre-migration distribution, flight patterns, and road traffic, we estimate that at least 61% (range 61% to 99.99%) of migrating monarchs are road-killed each fall. Although there is high uncertainty in our estimate, its magnitude suggests that roadkill could inhibit recovery of the population. Recovery planning should not only consider increasing the monarchs host plants, but must also address the reality of roadkill.

Diurnal ectotherms in hot and dry ecosystems face an acquisition trade-off between maximizing collective energy intake and minimizing individual exposure time. We employed optimal foraging and thermoregulation models to examine the surface foraging and thermoregulatory strategies of Pogonomyrmex barbatus foragers in response to habitat warming in a tropical deciduous forest. Foraging activity was evaluated using field-based thermal performance curves (fTPCs) for individual foraging speed and collective foraging success. Thermoregulation was assessed through indices of thermal unsuitability, as well as thermoregulatory effectiveness and exploitation. Our results show that afternoon conditions were significantly hotter and drier than those in the morning. Furthermore, the decoupling of fTPCs was dynamic; while the optimal temperature (Topt) for individual foraging speed remained stable at 44 degrees C, the Topt for collective foraging success shifted from 35 degrees C in the morning to 41 degrees C in the afternoon. This shift in foraging strategy, driven by stressful afternoon conditions, was reflected in a transition of the thermoregulatory strategy. Specifically, the probability of foraging activity decreased as thermal unsuitability increased, with a lower baseline in the afternoon. The thermoregulatory exploitation dropped from 94% in the morning to 63% in the afternoon. Based on thermoregulatory effectiveness, foragers transitioned from an active thermoregulatory strategy in the morning to a thermoconformist strategy in the afternoon while foraging on the surface. So, the observed colonies optimized the balance between individual foraging speed and collective foraging success through a thermoregulatory strategy that was primarily spatial in the morning and strictly temporal in the afternoon. A comparison with historical data from the same site reveals a 40% contraction in the total daily foraging window since 2013.

The red fox, Vulpes vulpes, is abundant in England and can exert limiting effects on their avian and mammalian prey. Large-scale gamebird releases in the UK may be sustaining high predator numbers, leading to greater predation on other prey species, especially once gamebird stocks are depleted. However, little is known how gamebird release affects the broader diet of the red fox. Here we used DNA metabarcoding to assess the diet of foxes from 18 agricultural estates in Southern England, 10 of which released large numbers of gamebirds (red-legged partridges - Alectoris rufus, and common pheasants - Phasianus colchicus) and 8 which did not. Scats were collected over one year, allowing for seasonal investigation of the foxes' diet. We investigated the vertebrate species consumed and compared the non-gamebird dietary diversity and composition between release and non-release estates and across seasons. The field vole (Microtus agrestis) was found to be the most frequently predated species overall. Brown hares and field voles were detected more on release sites, while bank voles and dog faeces were detected more on non-release sites. We found little evidence that foxes predate ground nesting birds or other species of concern. The dietary diversity was significantly lower on estates that released gamebirds, and this difference was most notable during the post-shoot, spring months (February to April). On both estate types, diversity was highest in the summer months. The altered predatory behaviour due to gamebird release is likely to affect the populations of the non-game prey of the red fox and this should be considered when designing policies regarding gamebird management and biodiversity conservation. Alternative predation combined with predator control may be reducing predation pressure on non-game prey where gamebirds are released, but high fox density elsewhere likely results in higher predation pressure on a wide range of species.

Benthic algae integrate catchment-and reach-scale pressures in streams. We assessed land-use effects on benthic algal assemblage structure, chlorophyll-a, ash-free dry mass (AFDM), and autotrophic index across 24 headwater streams in the Sondu-Miriu River basin, Kenya, and explored wet-dry patterns in two monitoring streams. We sampled across four land-use categories: natural forest (NF), tea and tree plantations (TTP), smallholder agriculture (SHA), and smallholder tea (SHT). Synoptic sampling occurred in August 2025, and one NF stream and one SHA stream were sampled five times between November 2024 and May 2025. Shannon and Simpson diversity were highest in NF streams and lowest in SHT streams. Chlorophyll-a and AFDM were highest in SHA streams; chlorophyll-a was lowest in SHT, while AFDM and autotrophic index were lowest in NF. Relative abundance patterns showed that SHA streams were dominated by Lyngbya and Stigeoclonium, whereas SHT streams were dominated mainly by Stigeoclonium; NF streams supported persistent diatom-rich assemblages. Non-metric multidimensional scaling showed land-use separation of benthic algal assemblages, while redundancy analysis indicated weak, non-significant evidence of environmental associations in the synoptic dataset. Composition and biomass metrics tracked land-use gradients and wet-dry patterns, supporting periphyton-based bioassessment for monitoring disturbance in Afrotropical headwater streams.

Climate change is thought to have altered the timing of avian migration, with species-specific responses varying across spatial and temporal scales. This is notable as ecological mismatches may occur when birds arrive or depart out of sync with the peak flowering period of their associated food plant resources. This study compares first arrival and last departure dates of Archilochus colubris (Ruby-throated Hummingbird) using citizen science data from the North American Bird Phenology Program for historical records (1880-1969) and recent records from eBird (2006-2024) and Journey North (2001-2024). Results show significantly earlier spring arrivals (14.2-27.9 days) and later fall departures (17.3-33.7 days) in the recent period compared to historical periods, while migration pace remained relatively consistent across datasets. Although temperatures are increasing in both the nonbreeding and breeding ranges of A. colubris, winter and fall temperatures were not strong independent predictors of migration timing. This study is the first to document shifts in fall departure dates for the species and highlights the need for continued research to uncover the mechanisms driving these phenological changes and assess their broader ecological implications.

Fluctuating asymmetry (FA) is widely used as a proxy for developmental instability, although reduced rather than increased FA, has occasionally been reported under particularly challenging conditions, possibly because environmental filtering removes less developmentally stable individuals. Here, we investigated temporal variation in wing shape asymmetry in males of Drosophila sturtevanti collected in 2022, 2024, and 2026 at the same sampling site, an urban Atlantic Forest fragment. Sampling in 2022 and 2026 were completed during single field expeditions in early April, whereas in 2024 repeated collections were required until mid-May because of markedly low capture success. We therefore examined whether this difference was associated with unusual environmental conditions and whether FA was higher in 2024, as expected under greater developmental perturbation. Daily temperature and relative humidity were examined from early March onward, encompassing approximately one month before adult emergence. The 2024 sampling period was warmer and drier than the corresponding periods in 2022 and 2026 and coincided with the final phase of the strong 2023-2024 El Ni&ntildeo event. Contrary to our prediction, FA was significantly lower in 2024. The magnitude of directional asymmetry remained similar among years, although the spatial pattern of the DA vector differed between 2024 and 2026. The coincidence of reduced capture success and reduced FA is consistent with the hypothesis that adverse conditions may impose demographic filtering, such that individuals expressing greater developmental instability are less likely to contribute to the sampled adult population.

Machine learning models can be used to analyze large bioacoustics datasets and explore variation due to geography or habitat. We find that in the monotypic Lark Sparrow (Chondestes grammacus), both environmental variables and geographic distance influence song variation in this species. Bird song is an important method of communication within avian species. The variation in bird song within a species can be due to a variety of factors, including genetic drift and isolation by distance. However, it remains unclear in species with wide ranges how environmental factors in particular can cause changes to the song. In this study, the song C. grammacus was analyzed via machine learning to determine if it had significant variation based on multiple geographical metrics. We trained a convolutional neural network to segment individual syllables of 91 C. grammacus recordings, then extracted song characteristics. We found that ecoregion and state explain variation in C. grammacus songs. Our results demonstrate the efficacy of using machine learning models to analyze large datasets, as well as the impact that ecogeographic variation has on song variance.

Smoke from spring boreal wildfires increasingly impacts eastern North America, exposing breeding birds to hazardous air pollution that may impact reproductive outcomes. Using data collected in 2018-2025 from 70,979 monitored nests of four widespread cavity-nesting songbirds, we found strong evidence that smoke greatly delays egg laying and can extend incubation and nestling duration. We further found that while smoke is associated with increased clutch sizes, in some species smoke exposure strongly decreases hatching or fledging success. Our results demonstrate that extreme smoke can have wide-ranging impacts on breeding birds, from altering phenology to impacting fitness. While the exact mechanisms underlying these results remain elusive, the full suite of effects suggests that modifications to adult behavior under smoky conditions is the most likely cause. As fire regimes shift, birds and other wildlife are at greater risk of exposure to toxic smoke during the breeding season, which may further exacerbate the biodiversity crisis.

The Alaskan Beaufort Sea lagoon system supports a diverse invertebrate community shaped by a complex glacial and biogeographic history, yet it remains severely understudied relative to the rapid environmental changes it is experiencing. As Arctic amplification accelerates habitat loss, shifts dispersal pathways, and facilitates the introduction of boreal and invasive species, accurately documenting current biodiversity is increasingly urgent. In this study, we used DNA barcoding of the mitochondrial cox1 gene for various invertebrates, and the nuclear 28S rRNA gene for Porifera, to examine invertebrate genera across coastal sites in the Alaskan Beaufort Sea, including the unique Boulder Patch kelp community in Stefansson Sound. We sequenced specimens belonging to five phyla: Annelida, Arthropoda, Mollusca, Porifera, and Priapulida. Specimens were identified by morphology, and their cox1 or 28S rRNA gene sequences were matched against the GenBank and Barcode of Life Data Systems (BOLD) databases. Molecular identification revealed substantially higher species diversity than morphology alone in three of five phyla: Annelida (12 versus six species), Arthropoda (14 versus 11), and Mollusca (five versus four). In Porifera and Priapulida the number of species were the same for morphological and molecular identification; however, we still found different species identifications between the two methods. Other notable findings include the discovery of multiple cryptic species within Terebellides sp., Pontoporeia femorata, Onisimus litoralis, and Micronephthys minuta, as well as the first published sequences for Acanthostepheia incarinata, Haliclona gracilis, Onisimus affinis, and Saduria sibirica. Our results highlight the extent to which morphology-based surveys underestimate invertebrate diversity in Arctic ecosystems and underscore the need for comprehensive surveys and molecular reference databases to support future molecular-based biodiversity monitoring in this rapidly changing region.

As one of the strongest selective forces, predation risk induces antipredator responses in prey that not only maximise survival probability but are energetically expensive. While these responses are known to affect foraging patterns and diet composition, the functional role of changes in dietary macronutrients remains unclear. We investigated the effect of dietary NPE (non-protein energy) concentration on the antipredator responses of the Indian rock agama Psammophilus dorsalis. In a manipulative experiment, wild-caught lizards were fed either a low or high NPE diet and exposed to repeated simulated predator attacks. We measured the space-use and hiding duration of lizards after each attack, as well as their circulating corticosterone concentrations at the end of the experiment. Predator exposure induced refuge use for all lizards. Diet had a context-dependent effect: lizards on the high NPE diet used riskier perch locations more frequently than those on low NPE diets, but only during safe periods and not during active threat. Physiologically, lizards on the low NPE diet had significantly higher baseline corticosterone levels, indicating a greater allostatic load. Overall, while dietary NPE did not alter reactive escape behaviour, it influenced the underlying physiological state and proactive behavioural decisions.

The coastal North Pacific kelp ecosystems are highly productive, supporting a rich faunal assemblage. Some historical aspects of this ecosystem remain poorly studied, as exemplified by the extinct Spectacled Cormorant Urile perspicillatus, whose Late Holocene distribution was apparently restricted to Bering Island. This cormorant's taxonomic relationships remain unresolved and little is known about its ecology. We use ancient-DNA to reconstruct the evolutionary history of the Spectacled Cormorant. Mitogenomes from five of the six known historical skins form a strongly supported clade within Urile cormorants. The ancestors of Spectacled Cormorant diverged from its closest relatives, the Pelagic (U. pelagicus) and Red-faced (U. urile) cormorants, ~6.98 Mya, and the population subsequently underwent a pronounced bottleneck. This divergence time is co-incident with the establishment of coastal kelp-forests. The Spectacled Cormorant may have been an integral part of the kelp-associated fauna as a large benthic-foraging predator. Similar to the sympatric Steller's sea cow (Hydrodamalis gigas), the historical relict distribution of the Spectacled Cormorant may be the result of trophic collapse of kelp-forest ecosystems and subsistence hunting. Future palaeogenomic and dietary isotopic research, and reassessment of North Pacific archaeological and fossil assemblages, should shed light on the evolution and palaeoecology of this enigmatic seabird.

Previously, we have demonstrated that certain ferric salts common in Archean waters, such as iron trichloride and ferric ammonium citrate, can protect liposomes from destruction by short-wavelength UVC light. In this study, we investigate the propagation of 254 nm UV radiation through aqueous FeCl3 solutions and its interactions with liposomes. We then consider these findings in the context of early Earth UV environment, discuss their implications for our hypothesis of the Darwinian evolution of liposomes, and integrate them with our previous experimental results.

1. Plants produce a great chemodiversity, which is the diversity of specialized metabolites (SMs). These SMs are produced in complex metabolic pathways and play an important role in inter-species interactions. There are numerous hypotheses about the evolutionary processes which brought about and maintain chemodiversity. Some have been partially tested in lab and field studies. However, some of their assumptions and predictions are better tested by quantitative modeling, and so far no quantitative model has investigated the role of metabolic pathways. 2. To close this gap, we developed an individual-based model for metabolic pathway evolution. It models enzymes creating metabolites with various modifications. Enzymes undergo inheritance and mutation. We used the model to compare the screening and interaction diversity hypotheses. 3. The screening hypothesis predicts promiscuous enzymes, genetic drift, the presence of many non-beneficial metabolites, and high metabolite richness. The interaction diversity hypothesis predicts specialized enzymes, selection, the almost exclusive presence of beneficial metabolites, and situation- dependent metabolite richness. We found that the patterns predicted by the screening hypothesis did not occur, while those predicted by the interaction diversity hypothesis did. 4. This provides reason to favor the interaction diversity hypothesis over the screening hypothesis when connecting empirical results to their evolutionary context

Copper is an essential micronutrient in most organisms that becomes toxic in large quantities. Repeated or prolonged sub-lethal exposure can lead to evolved resistance to copper toxicity over many generations, which may result in trade-offs between energetically expensive detoxification mechanisms and fitness. Alternatively, evolved resistance to chemical stressors may lead to correlated changes in other traits. This study focuses on a population of flies for which artificial selection for copper resistance led to an increase in both copper resistance and longevity. The apparent off-target benefit of copper selection on one component of fitness led us to investigate differences in fecundity and developmental viability in copper resistant and copper sensitive, non-selected populations. We assessed the effect of copper selection and copper exposure on multiple aspects of fecundity over the lifespans of females from the non-selected and copper-selected populations. Our study corroborated previously observed increased longevity in copper-selected flies. Controlling for variation in lifespan, copper-resistant females had comparable age-matched fecundity to copper-sensitive females and benefitted from increased longevity with higher lifetime fecundity. Overall, copper exposure negatively affected egg quality, but we found no difference in this trait between the copper-resistant and sensitive populations. Further, we found developmental viability under copper stress was significantly higher for eggs laid by copper-resistant females. Overall, we determined that copper resistant flies experienced a fitness benefit through both lifespan and fecundity. Costs of maintaining copper resistance may be associated with energetic costs, but these trade-offs may not always manifest in reproductive or lifespan fitness costs.

Tidal flats cover less than 1% of the surface of our planet, but they provide important ecosystem functions and services, like carbon sequestration and nutrient (re-)mineralization, primary, sec-ondary, and food production, and habitats for a variety of species. Despite their importance, they are threatened by environmental stressors and anthropogenic impacts. Hence, a better under-standing of the interaction between macrozoobenthos, their metabolic activity, and their contri-bution to carbon and nitrogen cycling is important. Therefore, we conducted in-situ pulse-chase tracer experiments at a Pacific oyster (Magallana gigas) reef in the Eastern Scheldt (Southwest Dutch Delta, Netherlands) in summer and autumn 2020. During these experiments, we provided the submerged benthic community with 13C and 15N-enriched bacterioplankton and incubated it in seawater enriched in 1% deuterium oxide (2H2O) for one tidal cycle (~9 h), before the benthic community was kept at in situ conditions for another 14 d. Our aim was (1) to assess seasonal variations in 13C and 15N processing by macrozoobenthos, (2) to evaluate potential life-stage de-pendent differences in 2H uptake - a proxy for metabolic activity, and (3) to identify the macro-zoobenthos taxon which had the highest metabolic turnover rate (MTR). In summer, most bacte-rioplankton-derived 13C- and 15N was taken up by sponges (Halichondria panicea), limpets (Crepidula fornicata), and clams (Ruditapes philippinarum), whereas in autumn, H. panicea, C. fornicata and the sponge Hymeniacidon perlevis dominated 13C and 15N incorporation. Statisti-cally significant differences in 2H uptake between life stages (juvenile/ small vs. adult) was de-tected in Malacostraca (i.e., Rhithropanopeus harrisii), but not in Polychaeta or Bivalvia. There-fore, this difference between taxonomic classes might be linked to mold-intermolt periods. Sea-sonal differences in MTR of H. panicea and M. gigas (i.e., the species with the highest MTRs in summer and autumn) might be linked to seasonal environmental variability, such as temperature and food availability.

Insects are a key group in ecosystems and economics, but are increasingly under threat, making ongoing monitoring essential to preserving them. Traditional monitoring approaches are time consuming, among other limitations. Automated acoustic recognition can be a good solution for monitoring arboreal, soniferous insects, such as cicadas, and the acoustic biology of cicadas in Australia is understudied, especially using automated methods. To evaluate the applicability of automated approaches for detecting cicadas, we used the deep-learning acoustic model BirdNET (v2.4) to perform embedding searches for Cystosoma schmeltzi (lesser bladder), Illyria burkei (eastern rattler), Macrotristria intersecta (corroboree cicada), Thopha sessiliba (northern double-drummer), and Pauropsalta opaca (fairy dust squawker), using audio recorded by Passive Acoustic Monitoring over 15-months at 18 sites in north Queensland. All classifiers achieved high area under the precision-recall curve (AUPRC) ranging from 0.753 to 0.982. We used a precision-prioritised threshold ([≥] 0.85) to constrain the false positive rate while retaining adequate outputs, which were consistent with the documented species ecology and life history. This approach was readily applicable to the Cicadidae, offering a scalable solution for biodiversity monitoring in complex acoustic environments.