Ecological Adaptation Research Articles

Comparative genomics reveals carbohydrate enzymatic fluctuations and herbivorous adaptations in arthropods

BackgroundArthropods represent the largest and most diverse phylum on Earth, playing a pivotal role in the biosphere. One key to their evolutionary success is their ability to feed on plant material. However, their endogenous enzymatic repertoire, which contributes to plant digestion, remains largely unexplored and poorly understood. ResultsWe analyzed 815 arthropod proteomes and identified a total of 268,171 carbohydrate-active modules. Our findings revealed a strong correlation between enzymatic content and feeding habits, with herbivorous species possessing significantly higher enzyme levels. We identified widespread carbohydrate-active families across the AA, CBM, GH, and GT classes, and observed a progressive increase in taxa-exclusive families in more recent arthropod lineages. Notably, we highlighted the impact of the transition from ametabolous to holometabolous development on carbohydrate metabolism, as well as the ecological adaptations of different species groups. By reconstructing the ancestral enzymatic profiles of arthropods, we identified significant fluctuations in 10 carbohydrate-active families over time. ConclusionsOur analysis advances the understanding of the evolutionary mechanisms utilized by the megadiverse phylum Arthropoda. We emphasize the critical role of herbivory as a selective force shaping enzymatic strategies, particularly those involved in carbohydrate metabolism. The distribution and exclusivity of carbohydrate-active families across different arthropod groups provide insights into their evolutionary trajectories and offer a clearer picture of the metabolic pathways that led their ancestors to their present forms.

A Global Synthesis of Environmental Enrichment Effect on Fish Stress

ABSTRACTThe stress‐coping ability (SCA) is one of the core aspects of fish welfare and is of vital importance for fish production in the aquaculture industry and for fish fitness in hatchery release. Environmental enrichment (EE), a method of introducing external stimuli into the husbandry environment, has been recently proposed to improve fish SCA, but the present experimental evidence is mixed, and the reasons for these discrepancies are unclear. Here, we conducted a global meta‐analysis using a data set that consists of 1786 cases from 107 studies across 42 fish species to solve this problem. Overall, we found that enriched fish had significantly higher SCA than control fish, reflected in either basal stress levels or stress responses. Meta‐regression analyses showed that specific subgroups of enrichment type, fish developmental stage, stress category, stress duration, stress place, sample tissue and indicator system showed significant positive EE effects on SCA. Multi‐model inference indicated that the indicator system, fish developmental stage, stress characteristic and enrichment mode are important drivers for the high heterogeneity among effect sizes. These results highlight the importance of introducing EE into the rearing systems, which will not only increase the welfare of aquaculture fish but also improve the ecological adaptability of released fish. The comprehensive knowledge obtained in this analysis will provide insights into fish ontogenetic plasticity and its responses to EE and have important implications for improving the production cycle in fish aquaculture and fisheries conservation.

Comparison of the Photosynthesis, Hydraulic Properties, and Anatomy of Pteroceltis tatarinowii Leaves Between a Limestone and a Cultivated Forest

Pteroceltis tatarinowii Maxim is a famous paper-making tree endemic to China with a wide distribution. Leaves of this tree growing in different habitats show a certain plasticity, which is important for their ecological adaption. Here, the photosynthesis ability, hydraulic properties, and anatomy of P. tatarinowii leaves from a limestone forest (Langya Mountain) and a cultivated forest (Xiaoling Village) in Anhui province were compared. The results showed that leaves from Xiaoling Village had higher net photosynthesis rate and hydraulic conductivity, which were closely related to their higher vein density, stomatal density and palisade tissue thickness than leaves from Langya Mountain. However, lower leaf water potentials at turgor loss point and at 50% loss of conductivity, as well as a higher leaf hardness, for Langya Mountain leaves indicated their higher hydraulic safety and drought resistance than those of leaves from Xiaoling Village. This study reveals a hydraulic trade-off between efficiency and safety for P. tatarinowii leaves growing in distinct habitats. Further studies should include more habitats and different vegetation communities to clarify the ecological adaption so as to provide a scientific basis for the protection of this species.

Molecular Innovations Shaping Beak Morphology in Birds.

The beak, a pivotal evolutionary trait characterized by high morphological diversity and plasticity, has enabled birds to survive mass extinction events and subsequently radiate into diverse ecological niches worldwide. This remarkable ecological adaptability underscores the importance of uncovering the molecular mechanisms shaping avian beak morphology, particularly benefiting from the rapidly advancing archives of genomics and epigenomics. We review the latest advancements in understanding how genetic and epigenetic innovations control or regulate beak development and drive beak morphological adaptation and diversification over the past two decades. We conclude with several recommendations for future endeavors, expanding to more bird lineages, with a focus on beak shape and the lower beak, and conducting functional experiments. By directing research efforts toward these aspects and integrating advanced omics techniques, the complex molecular mechanisms involved in avian beak evolution and morphogenesis will be deeply interpreted.

RXLR effector genes mediate regional adaptation of Phytophthora infestans

AbstractLocal adaptation has been a central theme of eco-evolutionary research for decades. It is generally assumed that plant pathogens are locally adapted due to their standing interactions with biotic and abiotic factors in the ecosystem. Effectors, secreted small proteins encoded by pathogens, play critical roles in host–pathogen interactions, by activating host genotype-specific resistance, suppressing plant immunity, and playing other functions. In this study, we investigated the potential involvement of RXLR effector genes in ecological adaptation by examining the simple sequence repeat (SSR), virulence, and effector profiles in Phytophthora infestans isolates collected from two geographic regions differing in ecological environments. Genotypic analyses with SSR markers and virulence assays showed that the pathogen from the two regions shared genetic background but differed in virulence spectrums. High-throughput sequencing and expression analysis of 24 selected P. infestans isolates further showed variations in the RXLR effector repertoire, ranging from 536 to 548 for each isolate and the expression of effector genes was highly associated with the accumulation of homologous sRNA. Regional specific alleles were detected at 94 RXLR effector genes, and a specific accumulation of homologous 25–26 nt sRNAs was found at 67 RXLR effector genes. Two of the regional specific RXLR effector genes were confirmed to be virulence factors. Taken together, these results suggest that genomic and epigenetic variations in RXLR effector genes contribute significantly to the ecological adaptation of P. infestans populations and that regional specific effector genes will help to understand the adaptive landscape of pathogens and efficient use of host resistance genes.

Assessing the Viability of Translocated Mongolian Dung Beetles (Gymnopleurus mopsus) for Ecological Restoration in Republic of Korea: An Analysis of Environmental Adaptability

This study investigates the reintroduction and ecological adaptation of the endangered dung beetle, Gymnopleurus mopsus, in South Korea, a region from which it has been absent since the 1970s. To facilitate this, we imported genetically identical populations of G. mopsus from Mongolia and embarked on a comprehensive restoration research project. A key focus of this endeavor was to evaluate the adaptability of these beetles to the local environment, an essential aspect of successful reintegration of species from foreign ecosystems. Under meticulously controlled field cage conditions, we conducted an in-depth monitoring of the life history traits of G. mopsus. This monitoring revealed that the adult beetles, which entered hibernation in September 2019, began emerging in stages from late April to May 2020. Following hibernation, we observed that the adults engaged in reproductive activities from late-May until early-August, with the emergence of the first-generation (F1) adults occurring from late-July to mid-September. This led to a notable tripling in population size, increasing from 34 to 109 individuals. The successful survival and reproductive behaviors of these Mongolian dung beetles in the climatic conditions of Korea suggest a promising potential for their adaptation when reintroduced into native habitats. We are now directing our efforts towards long-term monitoring, focusing on the survival and reproductive efficacy of these new generations, to further validate the success of this ecological restoration project.

Linking phylogenetic niche conservatism in bacterial communities in sorghum root compartments revealed by the Hongyingzi cultivar.

The root system harbours complex bacterial communities, which are critical for plant growth and health. Significant differences exist between bacterial communities in the root compartments; however, limited reports have explored their phylogenetic composition and niche conservatism in the root system of sorghum. We used the sorghum Hongyingzi cultivar as test plant, and applied 16S rRNA high-throughput sequencing and various statistical approaches. Phylogenetic composition of bacterial communities in root compartments were primarily driven by closely related species with similar environmental adaptations. We also found evidence of phylogenetic niche conservatism in bacterial communities for edaphic factors in the various root compartments, with pH and available N playing essential roles in shaping community composition. Environmental threshold analysis revealed threshold ranges of dominant taxa for pH and available N, indicating wider adaptive thresholds for more abundant taxa. Reconstruction of ancestral states suggested evolutionary changes in adaptability of certain bacterial taxa to edaphic factors, suggesting a shift towards slightly acidic, high N environments and reflecting the prolonged mutual interaction between bacteria and plants in cultivated soils. These findings enhance our understanding of environmental responses and evolutionary dynamics of root-associated microbiota in young sorghum plants and provide novel insights into ecological adaptations, shedding light on their responses to environmental factors. Our study contributes to a better understanding of the ecological dynamics of root-associated microbiota and offers analytical pathways for exploring the nutritional regulation of root microbiota.

High-resolution chromosome-level genome of Scylla paramamosain provides molecular insights into adaptive evolution in crabs

BackgroundEvolutionary adaptation drives organismal adjustments to environmental pressures, exemplified in the diverse morphological and ecological adaptations seen in Decapoda crustaceans, particularly brachyuran crabs. Crabs thrive in diverse ecosystems, from coral reefs to hydrothermal vents and terrestrial habitats. Despite their ecological importance, the genetic mechanisms underpinning their developmental processes, reproductive strategies, and nutrient acquisition remain poorly understood.ResultsHere, we report a comprehensive genomic analysis of the green mud crab Scylla paramamosain using ultralong sequencing technologies, achieving a high-quality chromosome-level assembly. The refined 1.21 Gb genome, with an impressive contig N50 of 11.45 Mb, offers a valuable genomic resource. The genome exhibits 33,662 protein-coding genes, enriched in various pathways related to development and environmental adaptation. Gene family analysis shows expansion in development-related pathways and contraction in metabolic pathways, indicating niche adaptations. Notably, investigation into Hox gene regulation sheds light on their role in pleopod development, with the Abd-A gene identified as a linchpin. Post-transcriptional regulation involving novel-miR1317 negatively regulates Abd-A levels. Furthermore, the potential role of fru gene in ovarian development and the identification of novel-miR35 as a regulator of Spfru2 add complexity to gene regulatory networks. Comparative functional analysis across Decapoda species reveals neo-functionalization of the elovl6 gene in the synthesis of long-chain polyunsaturated fatty acids (LC-PUFA), suggesting its importance in environmental adaptation.ConclusionsOur findings shed light on various aspects of crab biology, including genome sequencing, assembly, and annotation, as well as gene family expansion, contraction, and regulatory mechanisms governing crucial developmental processes such as metamorphosis, reproductive strategies, and fatty acid metabolism.

Leaf growth in third dimension: a perspective of leaf thickness from genetic regulation to ecophysiology.

Leaf thickness, the leaf growth in the third dimension as quantified by the distance between the adaxial and abaxial surface, is an indispensable aspect of leaf development. The fitness of a plant is strongly influenced by leaf thickness via modulation of major physiological processes, including photosynthesis and water use efficiency. The cellular basis of leaf thickness by alterations in either cell size or the number of cell layers is envisaged using Arabidopsis leaf thickness mutants, such as angustifolia (an) and rotundifolia (rot). Environmental factors coordinate with endogenous signaling mechanisms to exhibit leaf thickness plasticity. Plants growing in different ecological and environmental regimes show different leaf thickness attributes. However, genetic and molecular understandings of leaf thickness regulation remain largely limited. In this review, we highlight how cellular growth is transposed to fine-tune the leaf thickness via the integration of potential cues and molecular players. We further discuss the physiological significance of leaf thickness plasticity to the environmental cues that might serve as ecological adaptation enabling the plants to withstand future climatic conditions. Taken together, we seek to bridge the genetics and molecular biology of leaf thickness to its physiological significance so that leaf thickness can be systemically targeted in crop improvement programs.

Upper Palaeolithic fishing techniques: Insights from the engraved plaquettes of the Magdalenian site of Gönnersdorf, Germany.

The ~15,800 year-old Magdalenian site of Gönnersdorf, in Germany, has produced 406 engraved schist plaquettes which have been extensively studied in the past. The introduction of advanced imaging technologies, notably Reflectance Transformation Imaging (RTI), has now precipitated a re-evaluation of these artifacts, uncovering nuanced depictions of fishing practices previously unrecorded for the Upper Palaeolithic. Our investigation harnesses RTI to elucidate fine engraving details on the plaquettes, revealing depictions of fish and accompanying grid motifs. The analytical process enabled by RTI has exposed an intricate link between the grid patterns and fish figures, showing that they were a deliberate combination portraying the use of fishing nets. This discovery posits a significant departure from earlier interpretations of the site's iconography, which predominantly emphasized more naturalistic representations of fauna. Furthermore, these findings illuminate aspects of Magdalenian cultural praxis, suggesting that representations of aquatic life and fishing technologies were not merely utilitarian in nature but were embedded within a broader symbolic framework. This study enhances our comprehension of Magdalenian peoples' interaction with the aqueous milieu, revealing a sophisticated symbiosis between ecological adaptation and artistic expression.

The fact that bat wings and legs must evolve together impedes ecological adaptation.

The fact that bat wings and legs must evolve together impedes ecological adaptation.

Pollen and Leaf Micromorphological Characteristics of Spiny Almonds (Prunus subgenus Amygdalus) in Iran.

This study investigates the micromorphological characteristics of pollen grains and leaf epidermal cells from 20 accessions across four species of spiny almonds using scanning electron microscopy. Thirteen quantitative traits of pollen grains, including exine sculpturing, were analyzed alongside qualitative features such as shape, exine sculpture type, and aperture type. Additionally, four quantitative and five qualitative features of the leaf epidermis were examined, focusing on cuticular ornamentation patterns, types of epicuticular wax, and stomatal measurements. The pollen grains were found to be isopolar monads, radially symmetric, medium-sized, varying from prolate spheroidal to prolate, and exhibiting 3-colporate to 3-colpate structures. The exine sculpturing was generally striate with short and long ridges, with or without perforations, and could be classified into three types. Notably, in the Isfahan population of Prunus lycioides and the North Khorasan population of Prunus spinosissima, the exine sculpture types were distinctly different, being rugulate and reticulate, respectively. Multivariate statistical analysis identified equatorial diameter, colpus length and width, and ridge width as key diagnostic markers for species identification within spiny almonds. Principal component analysis and hierarchical clustering further highlighted the significance of stomatal length, cuticular ornamentation patterns, and epicuticular wax types in differentiating among taxa. We conclude that groupings recognized in recent classifications of the subgenus Amygdalus (spiny almonds) remain challenging to delineate solely based on palynological data, as diverse pollen types are present across different clades and subclades. Furthermore, micromorphological leaf traits proved valuable in distinguishing certain spiny almond taxa, and the traits of epidermal cells may reflect the ecological adaptations of spiny almond species.

Ethnoecology: The Community Adaptation Patterns of Forest Management in Grobogan Central Java

The aims of this research are to describe community adaptation patterns and determine community perceptions regarding forest environmental management. This qualitative research was studied using an ethnoecological approach with the research focus being the people of the Perhutani area of ​​Grobogan Regency. This research carried out the data collection through interviews, observation and documentation, with the main informant being Pesanggem as a forest land manager. This research produced the first findings in the form of community adaptation patterns in managing the environment covering three aspects, namely ecological adaptation, economic adaptation and social adaptation in the Pesanggem community in Gedangan Village, Wirosari District under the auspices of BKPH Sambirejo. 1) Ecological adaptation in Pesanggem showed the availability of well water which previously only relied on rainwater, then adjustments to various types of plants, as well as an understanding of prohibitions and sanctions for illegal activities in forest areas; 2) economic adaptation showed an increase in income for Pesanggem from forest land management; 3) social adaptation showed the existence of cooperation between members and other farmer groups within the LMDH forum. The second finding is that the public's perception in interpreting environmental management is that there are 5 values, namely the values ​​of wisdom, independence, religion, creativity and healthy living. Based on the research results, the author recommends that forest area communities be involved in managing and guarding forests for generational sustainability, and Perum Perhutani should invite partners from surrounding school institutions as a means of educating generations in joint forest management.

Understanding the ecological versatility of Tetracladium species in temperate forest soils.

Although Tetracladium species have traditionally been studied as aquatic saprotrophs, the growing number of metagenomic and metabarcoding reports detecting them in soil environments raises important questions about their ecological adaptability and versatility. We investigated the factors associated with the relative abundance, diversity and ecological dynamics of Tetracladium in temperate forest soils. Through amplicon sequencing of soil samples collected from 54 stands in six forest sites across the eastern United States, we identified 29 distinct Amplicon Sequence Variants (ASVs) representing Tetracladium, with large differences in relative abundance and small changes in ASV community composition among sites. Tetracladium richness was positively related to soil pH, soil temperature, total sulphur and silt content, and negatively related to plant litter quality, such as the lignin-to-nitrogen ratio and the lignocellulose index. Co-occurrence network analysis indicated negative relationships between Tetracladium and other abundant fungal groups, including ectomycorrhizal and arbuscular mycorrhizal fungi. Collectively, our findings highlight the ecological significance of Tetracladium in temperate forest soils and emphasize the importance of site-specific factors and microbial interactions in shaping their distribution patterns and ecological dynamics.

The macroecology of Mesozoic dinosaurs.

Dinosaurs thrived for over 160 million years in Mesozoic ecosystems, displaying diverse ecological and evolutionary adaptations. Their ecology was shaped by large-scale climatic and biogeographic changes, calling for a 'deep-time' macroecological investigation. These factors include temperature fluctuations and the break up of Pangaea, influencing species richness, ecological diversity and biogeographic history. Recent improvements in the dinosaur fossil record have enabled large-scale studies of their responses to tectonic, geographic and climatic shifts. Trends in species diversity, body size and reproductive traits can now be analysed using quantitative approaches like phylogenetic comparative methods, machine learning and Bayesian inference. These patterns sometimes align with, but also deviate from, first-order macroecological rules (e.g. species-area relationship, latitudinal biodiversity gradient, Bergmann's rule). Accurate reconstructions of palaeobiodiversity and niche partitioning require ongoing taxonomic revisions and detailed anatomical descriptions. Interdisciplinary research combining sedimentology, geochemistry and palaeoclimatology helps uncover the environmental conditions driving dinosaur adaptations. Fieldwork in under-sampled regions, particularly at latitudinal extremes, is crucial for understanding the spatial heterogeneity of dinosaur ecosystems across the planet. Open science initiatives and online databases play a key role in advancing this field, enriching our understanding of deep-time ecological processes, and offering new insights into dinosaur macroecology and its broader implications.

Evolutionary integration of forelimb and hindlimb proportions within the bat wing membrane inhibits ecological adaptation.

Bats and birds are defined by their convergent evolution of flight, hypothesized to require the modular decoupling of wing and leg evolution. Although a wealth of evidence supports this interpretation in birds, there has been no systematic attempt to identify modular organization in the bat limb skeleton. Here we present a phylogenetically representative and ecologically diverse collection of limb skeletal measurements from 111 extant bat species. We compare this dataset with a compendium of 149 bird species, known to exhibit modular evolution and anatomically regionalized skeletal adaptation. We demonstrate that, in contrast to birds, morphological diversification across crown bats is associated with strong trait integration both within and between the forelimb and hindlimb. Different regions of the bat limb skeleton adapt to accommodate variation in distinct ecological activities, with flight-style variety accommodated by adaptation of the distal wing, while the thumb and hindlimb play an important role facilitating adaptive responses to variation in roosting habits. We suggest that the wing membrane enforces evolutionary integration across the bat skeleton, highlighting that the evolution of the bat thumb is less correlated with the evolution of other limb bone proportions. We propose that strong limb integration inhibits bat adaptive responses, explaining their lower rates of phenotypic evolution and relatively homogeneous evolutionary dynamics in contrast to birds. Powered flight, enabled by the membranous wing, is therefore not only a key bat innovation but their defining inhibition.

Identifying Calmodulin and Calmodulin-like Protein Members in Canavalia rosea and Exploring Their Potential Roles in Abiotic Stress Tolerance.

Calmodulins (CaMs) and calmodulin-like proteins (CMLs) belong to families of calcium-sensors that act as calcium ion (Ca2+) signal-decoding proteins and regulate downstream target proteins. As a tropical halophyte, Canavalia rosea shows great resistance to multiple abiotic stresses, including high salinity/alkalinity, extreme drought, heat, and intense sunlight. However, investigations of calcium ion signal transduction involved in the stress responses of C. rosea are limited. The CaM and CML gene families have been identified and characterized in many other plant species. Nevertheless, there is limited available information about these genes in C. rosea. In this study, a bioinformatic analysis, including the gene structures, conserved protein domains, phylogenetic relationships, chromosome distribution, and gene synteny, was comprehensively performed to identify and characterize CrCaMs and CrCMLs. A spatio-temporal expression assay in different organs and environmental conditions was then conducted using the RNA sequencing technique. Additionally, several CrCaM and CrCML members were then cloned and functionally characterized using the yeast heterogeneous expression system, and some of them were found to change the tolerance of yeast to heat, salt, alkalinity, and high osmotic stresses. The results of this study provide a foundation for understanding the possible roles of the CrCaM and CrCML genes, especially for halophyte C. rosea's natural ecological adaptability for its native habitats. This study also provides a theoretical basis for further study of the physiological and biochemical functions of plant CaMs and CMLs that are involved in tolerance to multiple abiotic stresses.

A high-quality genome of the early diverging tychoplanktonic diatom Paralia guyana

The diatom Paralia guyana is a tychoplanktonic microalgal species that represents one of the early diverging diatoms. P. guyana can thrive in both planktonic and benthic habitats, making a significant contribution to the occurrence of red tide events. Although a dozen diatom genomes have been sequenced, the identity of the early diverging diatoms remains elusive. The understanding of the evolutionary clades and mechanisms of ecological adaptation in P. guyana is limited by the absence of a high-quality genome assembly. In this study, the first high-quality genome assembly for the early diverging diatom P. guyana was established using PacBio single molecular sequencing. The assembled genome has a size of 558.85 Mb, making it the largest diatom genome on record, with a contig N50 size of 26.06 Mb. A total of 27,121 protein-coding genes were predicted in the P. guyana genome, of which 22,904 predicted genes (84.45%) were functionally annotated. This data and analysis provide innovative genomic resources for tychoplanktonic microalgal species and shed light on the evolutionary origins of diatoms.

Effects of Climate Change on Temperature Sensitivity of Vegetation Growth in Huang-Huai-Hai Plain: Spatial–Temporal Dynamics and Ecological Adaptability

This paper explores the optimal temperature change in normalized difference vegetation index (NDVI) growth in the Huang-Huai-Hai Plain under the background of climate change, aiming to better cope with the impact of global warming on vegetation growth. The temporal and spatial variation characteristics of the NDVI and temperature factors were analyzed by using the NDVI, average temperature (Tavg), maximum temperature (Tmax) and minimum temperature (Tmin) datasets from 1982 to 2020. Through the Buishand U test and sliding slope detection, 1998 was determined to be a year of abrupt climate change. Furthermore, SHAP important feature analysis, a generalized additive model, correlation analysis, and other methods were used to identify the trend in the optimum temperature of vegetation growth before and after climate change. The results showed that (1) from 1982 to 2020, Tavg, Tmax, Tmin, and the NDVI in Huang-Huai-Hai Plain showed a significant upward trend. At the same time, the spatial distribution of these indicators shows the distribution characteristics of high in the south and low in the north. (2) The NDVI was positively correlated with Tavg, Tmax, and Tmin, and the correlation with Tmin was most significant. (3) The most suitable Tavg, Tmax, and Tmin intervals for vegetation growth were 20~30 °C, 25~35 °C, and 16~25 °C, respectively. (4) The optimum temperature range of vegetation growth was expanded after climate change, and the change rates of the Tavg and Tmax lower limits reached 24% and 25%, respectively, under the best condition of vegetation growth. (5) After abrupt climate change, the temperature suitable for vegetation growth increased significantly in the northern part of the plain but decreased slightly in the southern part.

Alternating carnivore and Neanderthal activities at Escoural Cave: insights from the taphonomic and machine learning analysis of leporid remains

Exploring the varied subsistence strategies and cave occupation patterns of Neanderthals is key to understanding their complex behaviors and ecological adaptations. Small game consumption, in particular, is considered a relevant indicator of their behavioral complexity. Rabbit assemblages from Pleistocene cave sites provide valuable insights into Neanderthal interactions with small prey and potential competition with carnivores. Here, we present the first detailed taphonomic analysis of faunal remains from Escoural Cave (Portugal), where a European rabbit (Oryctolagus cuniculus) assemblage was found alongside Middle Paleolithic stone tools and some macromammal remains. This study combines traditional zooarchaeological and taphonomic analysis of the rabbit remains with multivariate statistics and machine learning methods to establish the origin of the accumulation, and the implications for Neanderthal subsistence and cave use. Results from the taphonomic analysis show no evidence of human consumption but abundant evidence of small terrestrial carnivore activity, primarily from lynxes. This could indicate a sequential occurrence of Neanderthal and carnivore activities in the cave, with Neanderthal activities likely related to something other than rabbit consumption. Our study contributes to characterizing Iberian carnivore fossil accumulations and differentiating between faunal assemblages accumulated by carnivores and those by hominins. Additionally, we show that the use of machine learning analysis provides a robust and objective method for identifying and classifying taphonomic signatures, enhancing the accuracy and reliability of our interpretations. Future work will focus on analyzing additional faunal collections from both past and new excavations at Escoural, to test whether carnivores and Neanderthals focused on different prey types and how they shared the cave space.