Biology and Life Sciences

Abstract: Plant taxonomic knowledge contains a long tail of infrequently encountered names, diagnostic characters, and nomenclatural decisions, yet model reliability across this distribution remains unclear. We developed the Chinese-language PTTB-600, comprising 200 general, 300 ordinary specialized, and 100 long-tail fill-in questions, and evaluated 31 large language models (LLMs) or run modes under closed-book conditions without retrieval augmentation. The first author drafted the question bank and answer key; three coauthors with doctorates in plant taxonomy reviewed them independently. All models scored at least 197/200 on general questions, and 21 achieved full marks. The six highest-scoring models answered 291-297/300 ordinary specialized questions (97.0-99.0%) but achieved 63.0-90.0% accuracy on long-tail fill-in questions. Gemini 3.1 Pro Preview ranked first at 587/600; ranks two through six formed a closely spaced cluster with no significant adjacent differences after Holm correction. Across 11 within-family comparisons, thinking-mode runs yielded 20-65 additional correct answers, chiefly on specialized and fill-in tasks. Factual errors were uncommon in routine undergraduate content and concentrated in the generation of rare genus names, fine diagnostic distinctions, and alternative nomenclatural treatments. Top-performing LLMs can provide reliable support for routine teaching under instructor oversight, whereas long-tail identifications and nomenclatural decisions require verification against authoritative sources.

Abstract: Internal selection is a process of natural selection in which fitness differences between organisms are caused by factors internal to the organism and are invariant across environments. Internal selection has been most successfully applied to explain conserved features such as the phylotypic stage of gnathostomes. In this contribution, we introduce a form of internal selection that can drive evolutionary transformations rather than trait conservation, i.e. directional internal selection. The basic idea is that adaptive evolutionary change of one character can induce directional selection on another trait due to constraints internal to the organism. We provide a formal definition of this concept and discuss examples of directional internal selection as well as its implications.

Abstract: The longstanding dream of resurrecting dinosaurs faces formidable obstacles. DNA decays rapidly, making direct recovery impossible. Even if successful in their aims, and modern “synthetic” methods achieve a complete and functioning non-avian genome, they cannot recreate specific dinosaur species because at least some of the crucial genetic information is completely absent from living relatives. Alternatively, it is suggested here that microbial vectors once mediated horizontal gene transfer (HGT) between dinosaurs and the ancestors of extant species. While HGT appears rare, the ecological continuity between dinosaurs, microbes, and coexisting plants offers a faint but plausible route for fragments of dinosaur DNA to survived in extant species. Building on advances in paleogenomics, synthetic biology, and comparative genomics, we outline a multi-step strategy to search for, validate, and functionally test dinosaurian “genetic shrapnel” preserved in the “dark genome” of candidate organisms – particularly that of gymnosperms. If such material could be recovered, it might provide missing pieces needed to reconstruct extinct dinosaur species. Even if the goal of dinosaur de-extinction remains forever beyond reach, pursuing this line of research could yield transformative insights in evolutionary biology, synthetic biology, and conservation science — and offer a compelling narrative to engage the public.

Abstract:

The COVID-19 pandemic led to a dramatic increase in healthcare‑associated infections (HAIs) and antimicrobial resistance (AMR), particularly in intensive care units (ICUs). This study aimed to provide a comprehensive genetic characterisation of opportunistic pathogens (OPs) circulating in the ICUs of two infectious diseases hospitals repurposed for treating severe COVID‑19 patients. Whole‑genome sequencing (WGS) was performed on 175 isolates recovered from patients and the hospital environment (including personal protective equipment, PPE) between 2021 and 2023. The species composition included Klebsiella pneumoniae (n=53), Acinetobacter baumannii (n=60), Staphylococcus aureus (n=20), Enterococcus faecalis (n=20), Escherichia coli (n=10), Proteus mirabilis (n=5), Pseudomonas aeruginosa (n=4), Enterococcus faecium (n=2), and Klebsiella variicola (n=1). Bioinformatic analysis included multilocus sequence typing (MLST), core genome single‑nucleotide polymorphism (SNP) analysis, phylogenetic reconstruction, and in silico detection of resistance and virulence genes and plasmid replicons. Among K. pneumoniae isolates, the high‑risk clone ST512 (47.2%) carrying bla_KPC‑3, pmrB(R256G), gyrA(S83I), oqxAB, qacEdelta1 and fosA dominated and showed intra‑hospital spread (9 patients, 7 PPE, 9 surfaces). A. baumannii was represented by ST2 (40.0%, all bla_OXA‑23‑positive), ST78 (28.3%, bla_OXA‑72/90) and ST19 (3.3%). For E. coli, clonal complex CC14 (including ST1193) carried bla_CTX‑M‑15, marR(S3N) and fluoroquinolone resistance mutations. S. aureus isolates were assigned to CC15, CC8, CC22 (MRSA) and CC97. Resistance genes against up to nine antimicrobial classes were identified. Core genome SNP analysis confirmed direct transmission of K. pneumoniae (1–6 SNP differences) between patients and healthcare worker gloves. PPE was a major reservoir (68.4% of environmental isolates). High frequencies of IncF, rep5a/rep16, rep9 and IncC plasmids were detected in E. coli, S. aureus, E. faecalis and P. mirabilis, respectively. The study demonstrates the power of WGS for high‑resolution epidemiological surveillance, confirms the critical role of contaminated PPE in nosocomial transmission, and highlights the dominance of internationally spreading MDR clones in COVID‑19 ICUs.

Abstract: The moisture regime is one of the principal determinants of plant distribution; however, traditional phytoindication systems generally treat it as a single ecological gradient, despite its complex environmental controls. This study examined whether vegetation responses could be used to integrate climatic, edaphic, and topographic determinants of moisture availability into ecologically meaningful indicator scales. Species occurrence records for the flora of the Dnipropetrovsk and Zaporizhzhia regions (Ukraine) were combined with climatic, edaphic, and topographic predictors. Canonical correspondence analysis was employed to identify moisture-related vegetation gradients, which were subsequently modelled using precipitation, soil texture, topographic wetness, and their interactions. New species-level indicator values were derived and evaluated against traditional Didukh and Ellenberg scales using independent validation criteria and null-model randomisation tests. Two partially independent moisture gradients were identified. The dominant edaphic component was primarily associated with soil water-retention capacity and showed strong agreement with traditional phytoindication systems, whereas the second component reflected the topographic redistribution of water and provided complementary ecological information. Empirically calibrated indices integrating climatic water input, soil texture, and topographic wetness reproduced vegetation responses and enabled spatial mapping of ecologically relevant moisture conditions. These findings demonstrate that ecologically meaningful moisture regimes are multidimensional and cannot be adequately represented by a single gradient. The framework refines traditional phytoindication by separating complementary mechanisms controlling plant water availability and provides a basis for improved ecological assessment and climate-change projections.

Abstract:

The Lake Wales Ridge in central Florida, USA, is a biodiversity hotspot characterized by species-rich xeric scrub and sandhill communities with high levels of endemism. The endemic Florida scrub millipede, Floridobolus penneri, and the more cosmopolitan Smokey Oak millipede, Narceus gordanus, are sympatric detritivores inhabiting these sandy upland systems. Previous research identified the mite, Narceolaelaps gordanus, as a host-specific parasite of N. gordanus. In this study, we evaluated mite associations on sympatric millipedes across 20 sites on the Lake Wales Ridge and examined whether these associations were consistent with parasitism, commensalism, or phoresy. We found that Narceolaelaps gordanus occurred on both F. penneri and N. gordanus, thereby contradicting previous observations of host specificity. Mite abundance differed between millipede species, with N. gordanus supporting more mites per individual than F. penneri. Independent of host species, larger millipedes supported greater numbers of mites. Over a three-month observation period, we found no visible evidence of exoskeleton or tissue damage associated with mite presence. In laboratory assays, defensive secretions from F. penneri and N. gordanus neither attracted nor repelled mites, suggesting that short-range host chemical cues may not strongly influence mite location behavior. Collectively, our findings indicate that the association between Narceolaelaps gordanus and sympatric millipedes on the Lake Wales Ridge is structured more by shared habitat use and host size than by strict host specificity or overt parasitism. The observed associations are more consistent with opportunistic commensalism or phoretic dispersal in which millipedes function as transport hosts between patchily distributed detrital habitats. Further research integrating behavioral observations, experimental studies, and molecular approaches will be necessary to resolve the ecological and evolutionary nature of these mite–millipede associations.

Abstract: Interactions between plants and microorganisms have long been a central topic in biological research. Bacterial symbiosis on leaf surfaces represents a distinctive and mutually beneficial system within the phyllosphere microbiome. Leaf nodules are the visible manifestation of the symbiosis and confer ecological advantages to host plants by enhancing host resistance against pathogens and herbivores. It has been hypothesized that these advantages promote higher diversification rates in host lineages, but this remains unresolved. Ardisia subg. Crispardisia and its close relatives (Amblyanthopsis and Amblyanthus) within Primulaceae is a typical plant group with leaf-nodule symbiosis, making them an ideal system for testing this hypothesis. In this study, we conducted extensive sampling of “Ardisioids” (Ardisia and its allies), and reconstructed their phylogenetic relationships and evolutionary history using plastid genomes and nuclear datasets (nrDNA and genome-wide SNPs). We clarified the phylogenetic positions of several “Ardisioids” genera (e.g., Sadiria, Tapeinosperma, Amblyanthus, and Amblyanthopsis) and multiple subgenera within Ardisia. We further detected a rapid radiation during the middle Miocene in Ardisia and its allies. Notably, we found that the leaf nodulated clade appears to have originated during this period, approximately 11–8 Ma. BAMM analyses revealed elevated diversification rates in leaf-nodulated lineages, while HiSSE analyses indicated that leaf-nodule symbiosis might have increased speciation rates without significantly affecting extinction rates. These results provide strong evidence that leaf nodule symbiosis represents a key evolutionary innovation that has promoted diversification in Ardisia and its close relatives.

Abstract: Since 1993, vegetation monitoring of alpine plant communities (snow-bed grassland and alpine tundra) has been performed at the Velino-Duchessa LTER site in the Central Apennines (https://deims.org/12c79ecb-7890-4b75-9655-0883dacd8a29), through phytosociological relevés and abundance and cover estimation of the vascular flora at fine scale. Air and soil temperatures, rainfall, snowfall, snow cover persistence and soil chemistry have also been monitored, following the Long-Term Ecosystem Research network’s distinctive integrated and ecological approach. Separately for both plant communities, we tested changes in taxonomic diversity (species richness and Shannon diversity) and functional composition (proportion of life form, morpho-functional types, Grime’s strategies, root types and root depth) using descriptive statistics and generalized additive mixed models. A trend of variation forced by increasing drought and snow shortage was recorded in both plant communities, with 30% change per year in species composition. Whereas there was no significative change in the total number of species, some sensitive species completely disappeared and a number of invader species appeared, with an increase in more thermophilic and drought-tolerant species and a parallel decrease in more mesic and cryophilic species. The phenomena described may be linked to the observed climate change occurring during the last 100 years in the Apennines, consisting mainly of a strong reduction in the duration of snow cover and an increase in mean and minimum annual temperatures in the mountains.

Abstract:

Non-geniculate coralline red algae of the order Sporolithales are ecologically important calcium carbonate producers in tropical and subtropical seas globally; however, molecular surveys of this group in the Bay of Bengal (BoB) are entirely lacking, and no species of Sporolithon Heydrich has been documented from this embayment. Here we report the first occurrence of Sporolithon from Saint Martin Island (ca. 20°37′N, 92°19′E), the sole coral island of Bangladesh, at the northernmost margin of the BoB. Eight specimens encompassing free-living rhodoliths to encrusting growth forms were collected from intertidal and shallow subtidal habitats in March 2024. Species identification was assessed by combining scanning electron microscopy (SEM) images with a six-locus molecular dataset comprising three plastid-encoded markers (rbcL, psbA, and UPA), the mitochondrial cytochrome c oxidase subunit I 5′ region (COI-5P), and two nuclear-encoded ribosomal markers (LSU and SSU). Both maximum likelihood and Bayesian phylogenetic analyses resolved all BoB specimens within a well-supported clade of Sporolithon indopacificum Maneveldt, Gabrielson & Kangwe, with 0.00–0.59% rbcL and 0.00% psbA sequence divergence from type specimens from Tanzania and previously published sequences from Fiji, China, and Guam (USA). SEM confirmed morpho-anatomical features concordant with the original species description, including a free-living rhodolith habit, monomerous construction, and tetrasporangial compartments with apical pore plugs. This record extends the confirmed range of S. indopacificum into the northeastern Indian Ocean, represents the first documented Sporolithon occurrence in Bangladeshi waters, and highlights the value of combining SEM with multi-locus barcoding for coralline algal surveys in undersampled tropical regions.

Abstract: Phytotelmata are natural water-holding microhabitats that support diverse aquatic communities, including immature mosquitoes. We analyzed the diversity, composition, and interaction network structure of Culicidae associated with two bromeliad species, Aechmea distichantha and Vriesea friburgensis, in two localities of the Yungas ecoregion in northwestern Argentina: La Florida Provincial Reserve (LFPR) (Tucumán) and Isla de Cañas (Salta). Immature mosquitoes were sampled during 2017-2018, and community structure was evaluated using Hill numbers, nonparametric richness estimators, bipartite network analysis, and canonical correspondence analysis (CCA). A total of 12 mosquito species were identified, with richness ranging from 5 to 9 species depending on bromeliad species and locality, with the highest richness recorded in V. friburgensis in LFPR. Sampling completeness ranged from 96.8% to 100%. Bipartite network analysis showed moderate connectance (0.50), low modularity (Q = 0.188), intermediate nestedness (NODF = 46.94), and low global specialization (H2’=0.273), suggesting a relatively generalized interaction structure. Culex coronator and Cx. imitator dominated the assemblage, while other species exhibited more restricted associations. The CCA indicated that species composition was mainly associated with bromeliad height above ground, internal water temperature, sediment, and pH. These results highlight the importance of microhabitat heterogeneity in structuring mosquito assemblages in Neotropical phytotelmata.

Abstract: This paper proposes that a fundamental organizational trend in biological, cognitive, cultural, and technological evolution is the emergence of progressively more capable representation–interpretation systems. Across these domains, evolution has repeatedly generated systems in which representations acquire causal significance through increasingly sophisticated processes of interpretation rather than through their physical composition alone. The cumulative consequence is the progressive expansion of representational autonomy: the ability of representations to preserve their functional identity despite being instantiated in changing physical substrates.The emergence of RNA-based replication established reproducible molecular representations capable of persisting through material turnover. The genetic code differentiated representation from interpretation through a conventional coding relationship not grounded in physico-chemical similarity between codons and amino acids, but realized by a dedicated translational apparatus. Human language distributed representations across individuals and generations through socially learned symbolic systems. Contemporary artificial intelligence extends this trajectory by externalizing interpretive processes into technological systems capable of learning, generating, and transforming representations. Taken together, these transitions reveal a common evolutionary dynamic in which progressively more capable representation–interpretation systems enable representations to retain, transmit, and extend their causal influence despite continual changes in their physical instantiation. From this perspective, evolution can be understood not only as the diversification of biological forms but also as the progressive evolution of representation–interpretation systems, through which physical systems acquire an expanding capacity to model, preserve, and transform functionally significant aspects of themselves and their environment.

Abstract: Opuntia ficus-indica (L.) Mill. is a crassulacean acid metabolism (CAM) xerophyte whose intraspecific morphological diversity remains poorly docu-mented in Andean agroecosystems. This study characterized the phenotypic diversity of 55 accessions collected along an altitudinal gradient (1,381–2,758 m a.s.l.) in the dry inter-Andean valleys of northern Ecuador. Twenty FAO/CACTUSNET and UPOV descriptors were analyzed using principal component analysis (PCA), multiple correspondence analysis (MCA), and Ward–Gower hierar-chical clustering. Quantitative traits showed moderate to high variability, with repro-ductive and defensive characters exhibiting greater variation than vegetative traits. MCA revealed substantial chromatic diversity, with fruit peel and pulp color emerging as the strongest qualitative discriminators among morphotypes. Cluster analysis iden-tified three statistically distinct groups: a small-fruited, highly spinescent morphotype restricted to the most arid sites; an intermediate and phenotypically diverse mor-photype distributed across all provinces; and a large-fruited, low-spinescence mor-photype with the highest seed numbers, consistent with a more advanced domestica-tion trajectory. The primary differentiation axis reflected a trade-off between spine in-vestment and reproductive output. Five discriminant descriptors—fruit weight, spine length, seed number, peel color, and pulp color—provide a robust baseline for germplasm characterization and support conservation, breeding, and nutraceutical valorization in Andean drylands.

Abstract: We examined the endolichenic fungal communities from three lichen species occupying the rocks in geographically and climatically different Israeli zones—the continental northern region, the Mediterranean area, and the central Negev Desert. Overall, 103 fungal species belonging to 61 genera were isolated from the lichens collected during three seasons. The majority of fungi were melanin-containing: 69 species, comprising 76.8% of all isolates. The relative abundances of melanized species with thick-walled and multicellular spores in each season were substantially higher in the most climatically severe desert regions. Seasonal effect was significantly expressed on species richness and isolate densities of the endolichenic communities, which were the lowest in the summer. Both these characteristics were also significantly dependent on lichen species hosted the communities, and were the lowest in the very thin thalli of Circinaria calcarea. Fungal communities from the nearby rock surface formed more coherent groups, and their complexes of dominant species were subjected to clearer regional and seasonal variations in comparison with the endolichenic communities. This fact is explained by the specific conditions of lichen hosts possessing the differences in growth form, moisture regime, and metabolic activity, which can substantially affect the formation of endolichenic communities.

Abstract: How vertebrate nervous systems establish stable spatial reference frames remains a central question in comparative neuroscience, vestibular biology, and embodied cognition. Modern neuroscience explains upright perception through multisensory calibration rather than the literal inversion of retinal images, yet the evolutionary origins of gravity-anchored spatial organization remains incompletely understood. This paper proposes a comparative evolutionary hypothesis: terrestrialization progressively increased selective pressure for stable gravity-referenced multisensory integration, favoring tighter coordination among vestibular, visual, proprioceptive, and postural systems. The framework does not claim that gravity uniquely determines perception, nor that all vertebrate spatial organization derives solely from terrestrial locomotion. Instead, it proposes that gravitational vertical became an evolutionarily privileged calibration scaffold under conditions of sustained weight-bearing instability and substrate-dependent locomotion. Evidence is reviewed across aquatic, amphibious, terrestrial, arboreal, aerial, and secondarily aquatic vertebrates. Natural comparative cases are interpreted as evolutionary perturbation tests rather than linear progressions. The framework is intentionally comparative and phenomenological: it offers a structured evolutionary scaffold that generates operational predictions concerning vestibular organization, altered-gravity adaptation, developmental plasticity, and comparative neuroanatomy.

Abstract: Since its inception in the 19th century, the concept of ecology has continuously expanded in scope—from individual organisms to the biosphere—and deepened in focus, shifting from organism-environment interactions to the structures and functions of ecosystems. As an interdisciplinary concept prone to controversy and revision, ecology plays a pivotal role in advancing ecological civilization. However, the absence of a mechanism-based and time-appropriate definition has hindered the integration of cross-hierarchical research and the response to practical demands amid global environmental changes. This paper briefly reviews the origin and evolutionary context of the ecology concept and identifies the limitations of previous definitions. It then elaborates on the essence of ecology, and proposes an updated redefinition: Ecology explores the structures and functions (material cycling and energy flow) of the bio-hierarchy (total ecosystems), which is co-shaped by information-mediated feedback regulation between organisms and their biotic and abiotic environments, self-organization and structuralization, as well as human-nature coupling, and provides theoretical foundation and practical solutions for the harmonious coexistence between humanity and nature. Finally, the paper outlines future research directions centered on establishing a unified, cross-hierarchical ecological theoretical framework to support ecological civilization construction and global eco-environmental governance.

Abstract: Benthic macroinvertebrates are widely used as bioindicators for assessing freshwater eco-system health. This study investigated the diversity patterns and community structure of benthic macroinvertebrates across 21 sampling sites along the middle and lower reaches of the Yangtze River. A total of 74 species belonging to 3 phyla, 7 classes, 17 orders, 37 families, and 58 genera were identified, with aquatic insects dominating the assemblages. Alpha diversity indices showed no significant differences among river sections, whereas multivariate analyses (NMDS and PERMANOVA) revealed significant spatial variation in community composition, indicating that beta diversity plays a key role in structuring as-semblages at the basin scale. Canonical correspondence analysis (CCA) identified nutrient variables (TN and NH₄⁺-N), as well as pH and conductivity, as the main environmental drivers influencing community distribution. The results suggest that benthic macroinver-tebrate diversity patterns in large river systems are jointly shaped by regional environ-mental gradients and local habitat conditions. These findings provide insights into biodi-versity conservation and ecological management of large river ecosystems.

Abstract: The "RNA World" hypothesis posits a reasonable scenario for the origin of life. A question central to this scenario is: which RNA species may have emerged as the first ribozyme, thereby initiating the early life stage? To date, experimental and computational studies focus on ribozymes playing a constructive role in RNA synthesis—such as those facilitating template-directed copying or nucleotide synthesis. However, the answer remains unconvincing, primarily because their functional complexity likely demands lengthy sequences, making de novo emergence improbable. Here, we propose an alternative scenario: a destructive ribozyme, which is potentially quite short, may have emerged first. This ribozyme could cleave other RNA molecules to generate building blocks for its own replication. Notably, the dilemma for a destructive ribozyme is that it may degrade molecules of its own type, thus making it probably difficult to thrive as an RNA species. Using computational modeling, we demonstrate that an exonuclease ribozyme encoded within a circular RNA genome (thus evading self-attack) can spread within an RNA pool and may subsequently give rise to those constructive ribozymes. Our findings offer a new perspective on the initiation of the RNA world, thereby enriching both theoretical and empirical frameworks for understanding the origin of life.

Abstract: Downstream migration (DSM) of young fish is an efficient way of their dispersal in rivers. Regulation of rivers transform them into lotic-limnetic cascades fragmented by dams. Novel conditions of transformed riverine habitats influence migration routes and behavior of migrants, creating hazardous ecological situations, which lead to enhanced mortality and reduced fitness of fish. The review is aimed at ecological effects of river regulations on DSM, taking into account not only entrainment, damage and mortality of migrants in the water abstraction systems at the dam, but also modifications of DSM in the whole reservoir and within its sections. Habitat heterogeneity/complexity is one of the key factors controlling DSM both in natural and regulated rivers. It influences foraging, defense, migratory behavior at different spatial scales. In reservoirs, topography of the whole reservoir, stretched hydraulic gradients between migratory and residence habitats, widen inshore areas rich of shelters and landmarks provide higher retaining capacities for migrating fish. Homogeneous open waters are less suitable for feeding, communication, defense, and other behaviors of migrants than heterogeneous inshore habitats. Three ecologically distinct zones were recognized in the reservoir, where specific modifications of DSM were described. Fish from different taxonomical and ecological groups, e.g. Cyprinidae and Percidae, respond differently to the newly emerged structures and modified habitat heterogeneity. When creating fish-friendly migration routes and developing conservation measures, we have to take into account not only transformed habitats, but also characteristics of different ecological groups of fish.

Abstract: Aspergillus fumigatus is the predominant pathogenic fungus responsible for aspergillosis. In recent years, the global detection rate of azole-resistant A. fumigatus (ARAF) has continuously increased, and the extensive application of agricultural azole fungicides has been recognized as a crucial driving factor for the emergence and spread of resistance mutations in environmental A. fumigatus. Previous investigations conducted by our research team in karst vegetable fields of Guizhou Province revealed that the resistance rate of local A. fumigatus was only 0.49%, which was markedly lower than those reported in most previous studies in China and outside of China. To supplement the prevalence data of azole resistance across different habitats in this region, a total of 191 environmental A. fumigatus strains were isolated from nine tea plantations across Guizhou. In this study, two clinically prevalent azole drugs, itraconazole and voriconazole, were used for antifungal susceptibility testing, and the triazole target gene cyp51A of all isolates was sequenced and analyzed. Antifungal susceptibility results demonstrated that the MIC ranges of the tea plantation A. fumigatus population were 0.015–0.5 μg/ml for itraconazole and 0.031–0.25 μg/ml for voriconazole, with no evidence of triazole resistance. Genetic analysis identified ten different gene mutations among 29 isolates, all of which were classified as non-resistance-associated mutations. Among these mutations, four were synonymous mutations, including 267G→A, 540G→A, 1074A→G, and 1362T→C, while six were non-synonymous mutations, including 137T→A, 514A→G, 743A→C, 744T→A, 765C→G, and 1279G→A. These non-synonymous mutations resulted in five amino acid substitutions in 25 strains, namely F46Y, M172V, N248T/K, D255E, and E427K. The N248T/K mutation exhibited the highest mutational frequency of 0.1309 (25/191) and was distributed across all sampling sites. Correlation analyses indicated that no significant correlations were observed between all detected variant loci and MICs of isolates to itraconazole and voriconazole. Phylogenetic analysis revealed that the six sequence types of cyp51A in Guizhou tea plantations were broadly intermixed with those from other parts of China and outside of China. We discussed the implications of these results in the management of ARAF.

Abstract: Rising global temperatures pose a serious threat to wild small mammal’s population persistence. In this study, we investigated the molecular mechanisms underlying heat-induced testicular impairment in Brandt’s vole (Lasiopodomys brandtii), a dominant small mammal species of the Eurasian temperate steppe. Adult males were subjected to short term heat exposure at 37°C, 39°C, and 41°C. Heat stress at temperatures ≥39°C significantly reduced the testicular index and caused histopathological damage. Integrated transcriptomic and data-independent acquisition proteomic analyses revealed significant enrichment of pathways related to endoplasmic reticulum protein processing and barrier function. Further molecular validation demonstrated robust activation of the unfolded protein response, indicated by increased expression of ATF4, ATF6B, phosphorylated eIF2α, and XBP1. Together, these results identify endoplasmic reticulum stress as a key mediator of heat-induced testicular injury and highlight that 39°C represents a critical reproductive threshold for Brandt’s voles, even following short-term exposure.