Biology subjects

Deng, Z.

Publications and source records attributed to Deng, Z..

At least 37 records · Page 2Linked to original sources

bioRxiv2025

From Hands to Feet: Experience-Driven Plasticity in Secondary Somatosensory Cortex in People Born without Hands

In congenital handlessness, where individuals rely on their feet for compensatory manual functions, reorganization in the primary somatosensory cortex (SI) appears limited to local shifts by adjacent body parts. It remains unclear how congenital handlessness and lifelong compensatory foot use shape the sensorimotor system beyond SI, and whether compensatory abilities can drive broader functional reorganization. Here, we used task-based and resting-state functional magnetic resonance imaging (fMRI) to investigate how compensatory foot use changes body representations in the secondary somatosensory cortex (SII) in people born without hands (individuals with upper-limb dysplasia, IDs). We found that IDs showed foot selectivity in the left hand-selective SII, with stronger foot activity than typically-developed (TD) control participants during action execution. Notably, this reorganization in the left SII hand area cannot simply be explained by this areas intrinsic organization - the typical secondary preference of this area in the TDs when hand responses are excluded, as this area in TDs shows shoulder - but not foot - selectivity when the hand activity is excluded. This indicates a functional reorganization dependent on the IDs sensorimotor experience. Resting-state results further revealed altered functional connectivity between SII and primary sensorimotor regions in IDs. Our findings suggest that SII exhibits flexible, experience-dependent plasticity, revealing a hierarchical principle of cortical plasticity, whereby reorganization in the SI is constrained by somatotopic principles while SII reflects functionally or experience-dependent plasticity.

neuroscience↗

bioRxiv2025

Distinct Programs Drive Organ Development and Regeneration

A prevailing paradigm in stem cell and developmental biology posits that regeneration is a recapitulation of embryonic programs, yet the molecular logic underlying these processes remains unclear. Here, using the mammary gland as a model, we identified a novel intermediate cell state essential for basal-to-luminal differentiation during regeneration. The transcription factor Pou2f3 is a master regulator of this state, critical for regeneration but dispensable for organ physiology. Mechanistically, we found that POU2F3 functions as a transcriptional repressor that drives lineage conversion by restricting chromatin accessibility of basal genes rather than by directly activating luminal programs. Furthermore, we uncovered a POU2F3-TNF feedback loop that maintains epithelial homeostasis, where TNF signaling suppresses Pou2f3 to inhibit differentiation, and loss of luminal cells relieves this repression to trigger regeneration. Strikingly, cell fate is governed by signaling pathways essential for cell-cell communication during development, but by Pou2f3-dependent chromatin remodeling for rapid response and injury repair. We validate the regeneration-specific requirement for Pou2f3 in mammary gland and further demonstrate analogous regeneration defects in prostate and pancreas. These findings challenge the long-held assumption that regeneration recapitulates development and establish a framework for understanding context-specific mechanisms of tissue plasticity, with implications for regenerative medicine and epithelial biology.

cell biology↗

bioRxiv2025

LRP8 is a Receptor for Yellow Fever Virus

Yellow fever virus (YFV), an arbovirus causing substantial human morbidity and mortality, was the first human virus discovered over a century ago. The live-attenuated 17D vaccine is among the most successful vaccines in medicine. Despite the importance of YFV, its receptor has remained unknown. Here, we performed a barcoded, genome-wide human ORF library screen and identified LRP8 (also named APOER2) as a receptor for YFV. We show that LRP8 expression specifically boosts YFV infection in cell lines by promoting entry. AAV-mediated expression of human LRP8 in mouse liver aggravates infection and pathology. LRP8 knockdown abolishes YFV infection in brain cells, primary human hepatocytes, and notably in mosquitoes. Biochemically, LRP8 directly interacts with YFV particles via the viral envelope protein. This function of LRP8 is conserved across species, particularly in mosquitoes and primates. A soluble LRP8 decoy protein can block YFV infection in vitro and in mice, providing a potential therapeutic or prophylactic strategy. Our findings provide groundwork for understanding YFV entry, tropism, and pathogenesis, and may enable development of novel therapeutics to treat YFV infection.

microbiology↗

bioRxiv2025

Structure and function of the nairovirus cap-snatching endonuclease

Nairoviruses include several human pathogens such as Crimean-Congo hemorrhagic fever virus (CCHFV) and Kasokero virus (KASV). The cap-snatching endonuclease (EN) domain of the viral polymerase is essential for transcription and represents a promising antiviral target. However, the structural and functional mechanisms of nairovirus ENs remain poorly understood. Here, we describe biochemical and structural studies of the ENs from CCHFV and KASV. Biochemical assays demonstrate that the RNA endonuclease activity of both ENs is activated by manganese ions and exhibits a preference for uridine-rich RNA substrates. This activity is inhibited by three metal-chelating inhibitors (DPBA, L-742,001, and BXA), with BXA displaying the highest binding affinity and inhibitory potency. We further determine nine crystal structures of CCHFV and KASV ENs in apo, metal ion-bound, and inhibitor-bound states. Comparative structural analysis uncovers a two-metal-ion binding mode unique to nairovirus ENs, in which conserved residues coordinate two manganese ions via bridging water molecules. In the inhibitor-bound structures of KASV EN, BXA forms additional stabilizing interactions with the enzyme, explaining its superior inhibitory effect. Functional assays further confirm that the two-metal-ion mechanism is critical for viral transcription. These findings provide a structural foundation for the rational design of antivirals against CCHFV and related pathogens.

microbiology↗

bioRxiv2025

RNA viruses in water shape the viromes of shrimp and fish in aquaculture systems

Aquatic environments host immense RNA viral diversity, yet the extent to which waterborne viruses influence the virome of aquatic animals remains unclear. Here, we conducted a comprehensive longitudinal metatranscriptomic analysis of water, shrimp, and fish across multiple culture stages in aquaculture systems. We identified 3,211 RNA viruses spanning 21 viral supergroups, with 97.4% representing novel species. Water exhibited the greatest viral diversity (3,093 species), followed by shrimp (1,280) and fish (398), with most viruses detected in shrimp and fish also detected in water. Diversity analyses revealed increasing richness and significant compositional shifts in 92 high-abundance viruses (>1,000 RPM) across culture stages. Viral abundance in aquatic animals was strongly correlated with that in water, and the temporal trajectories of the 16 most abundant viruses were closely synchronized between water and shrimp. Furthermore, 337 virus species were shared among shrimp and two fish species, of which 309 were present in water. These findings demonstrated that RNA viruses in water shape the composition and dynamics of aquatic animal viromes, underscoring the importance of water virome surveillance and management for disease prevention in shrimp aquaculture.

microbiology↗

bioRxiv2025

Crosstalk between Immune Microenvironment and Hair Follicle Cells Underlies Sexual Dimorphism in Androgenetic Alopecia

Androgenetic alopecia (AGA), also known as female pattern hair loss (FPHL) in women, is the most prevalent form of hair loss. It is characterized by progressive miniaturization of hair follicles and shortening of the anagen phase. The condition is widely attributed to genetic predisposition and androgen-mediated activation of androgen receptors. Other factors, such as the immune microenvironment, could also contribute to the pathogenesis. However, the specific mechanisms involved are still poorly understood. This study aimed to investigate the potential role of skin-resident myeloid cells in interacting with hair follicle cells under androgen stimulation, and to elucidate the sex-specific differences in dihydrotestosterone (DHT)-induced hair loss. Both female and male mice received DHT treatment, and histological examination was performed to assess DHT-induced alterations in skin morphology. Single-cell RNA sequencing (scRNA-seq) was utilized to profile skin cell populations and explore underlying mechanisms. Our results demonstrated that DHT inhibited hair regrowth in both sexes, altered skin thickness, and induced hair follicle miniaturization. scRNA-seq analysis revealed enhanced interactions between myeloid and fibroblast subpopulations, with more robust crosstalk observed in female mice. An in vitro experiment demonstrated that DHT promoted apoptosis of dermal papilla cells (DPCs) in the presence of macrophages. Treatment with polypeptides Y001 and Y002 effectively promoted hair regrowth by suppressing apoptosis signaling pathways. Our findings underscore the interactions between immune cells and hair follicular cells, particularly mediated by skin-resident myeloid cells, in the sexual dimorphism of androgenetic alopecia. The polypeptides Y001 and Y002 exhibit promising therapeutic potential by targeting apoptotic pathways, offering novel avenues for AGA treatment.

cell biology↗

bioRxiv2025

Discovery of circulating cell-free DNA 5hmC biomarkers for peritoneal metastasis in colorectal and appendiceal cancer

IntroductionPeritoneal metastases (PM) are associated with poor prognosis in patients with colorectal cancer (CRC) or appendiceal adenocarcinoma (AA), yet detection of PM is unreliable using current circulating DNA technology. Leveraging novel 5hmC-seal technology to detect ultra-low amounts of DNA in plasma, we demonstrate the feasibility of 5-hydroxymethylcytosine (5hmC) signatures derived from circulating cell-free DNA (cfDNA) as biomarkers for PM. MethodsUsing a highly sensitive and robust 5hmC sequencing approach on genomic DNA isolated from peripheral blood samples, we developed predictive models to identify biomarkers for peritoneal metastases. ResultsWe obtained genome-wide 5hmC profiles from 71 CRC/AA patients with PM, 41 without PM, and 73 non-cancer controls. Predictive models trained on genomic region 5hmC levels in patients with cancer could distinguish PM status with high sensitivity and moderate specificity (AUC 0.827, sensitivity 92.4%, specificity 46.1%). Pathway enrichment analysis identified epigenetically dysregulated cancer, cell migration, adhesion, and immune-related pathways in PM. ConclusionNovel 5hmC-Seal technology based 5hmC signatures can detect patients with peritoneal metastases from CRC and AA, albeit with reduced specificity. This study lays a foundation for future clinical assay development for PM. Statement of significanceWe demonstrate high-sensitivity detection of peritoneal metastasis in colorectal and appendiceal adenocarcinomas using 5hmC-Seal of plasma cfDNA. Earlier detection of this condition could expand curative treatments in [~]20,000 affected U.S. patients.

cancer biology↗

bioRxiv2025

DynPerturb:Dynamic Perturbation Modeling for Spatiotemporal Single-Cell Systems

Perturbation responses in multicellular systems are inherently spatiotemporal, governed by temporal dynamics, perturbation intensity, and tissue context. However, existing approaches rarely capture explicit spatiotemporal perturbation dynamics, limiting the ability to reconstruct how perturbations propagate over time, spread across space, and induce feedback at multiple biological scales. Here we introduce DynPerturb, a dynamic inference framework for single-cell and spatial transcriptomics that systematically models the "when-how strong-where" dimensions of perturbations. DynPerturb incorporates temporal encoding, spatial adjacency, and memory modules to capture nonlinear signal propagation and cross-regional feedback with mechanistic interpretability. Across temporal benchmarking datasets, DynPerturb consistently outperformed existing methods in regulatory link prediction, demonstrated zero-shot generalization to unseen nodes, and preserved accuracy under varying sparsity conditions and temporal resolutions. Applied to representative biological systems, DynPerturb uncovers dynamic regulatory rewiring in kidney disease, highlighting a mid-age sensitivity window and potential reversibility with early intervention; delineated a temporal fate-switching boundary between megakaryocyte-erythroid progenitors (MEPs) and granulocyte-monocyte progenitors (GMPs) during hematopoiesis; and revealed spatiotemporal interchamber signaling in murine heart development, where Igf2 perturbations induced spatial feedback shaping chamber proportioning. Collectively, these results establish DynPerturb as a unified and versatile system for dissecting dynamic perturbation effects and guiding precision intervention strategies in multicellular systems.

bioinformatics↗

bioRxiv2025

Phosphate limitation triggers Fe3+-dependent polymyxin resistance in Enterobacteriaceae

Phosphate (Pi) scarcity, a pervasive stressor prevalent in bacterial infections and dysbiotic host environments, potently drives antimicrobial resistance (AMR) against cationic antibiotics like polymyxins across diverse bacterial taxa. While established Pi depletion-induced AMR mechanisms often involve membrane phospholipid remodelling, Enterobacteriaceae largely lack these pathways, leaving their Pi scarcity-driven AMR mechanism unresolved. Here, we reveal that Pi limitation robustly induces polymyxin resistance in Enterobacteriaceae through 4-amino-4-deoxy-L-arabinose (L-Ara4N) modification of the Gram-negative outer membrane component lipid A, reflecting a distinct adaptive strategy. This modification is driven by strong ugd-arn operon induction, mediated by the PmrAB two-component system. We discover that Pi depletion triggers cellular Mg{superscript 2} release, prompting compensatory Fe3 mobilization to the cell envelope that directly activates PmrAB. Crucially, this metal-dependent signaling axis offers a directly targetable mechanism for reversing polymyxin resistance, a significant deviation from the less accessible, PhoBR-regulated phospholipid remodelling strategies in other bacteria. We demonstrate that Mg{superscript 2} supplementation or Fe3 chelation effectively suppresses PmrAB activation and restores polymyxin susceptibility, thereby establishing a novel, metal-centric paradigm for understanding and pharmacological intervention in stress-induced AMR.

microbiology↗

bioRxiv2025

Spatial Transcriptomics Reveal Developmental Dynamics of the Human Cerebral Cortex and Striatum

The human fetal brain undergoes morphological changes that contribute to the development of regional functionalities. However, the features of structural development, the underlying molecular and cellular signatures in the fetal brain remain unclear. With spatial transcriptomics and snRNA-seq, we identified 25 forebrain regions and characterized the dynamic changes in the cortex and striatum during the late first and early second trimesters. In particular, we discovered that temporal lobe enriched NPY-expressing L2/3 EX neuron potentially interacted with L4 EX neurons during cortical expansion and arealization. Additionally, the gyrus and sulcus were developmental asynchronous, in which HOPX and SPARC genes were potentially involved. Further investigation on the striatum showed specific genes and cell types that enriched in patch and matrix compartments, and SST-positive interneurons potentially involved in the development of these structures. Together, our results give insights into the understanding of early fetal brain development.

bioinformatics↗

bioRxiv2025

Comparative analysis of naked mole-rat thermogenesis and its potential to maintain euthermia in response to cold

The naked mole-rat (NMR) is a subterranean rodent known for its unique thermal biology, exceptional longevity and resistance to cancer and hypoxia. However, its thermal biology remains controversial, with various reports describing NMRs as poikilotherms, heterotherms, mesotherms or partial homeotherms. Here, we investigated whether the thermogenic potential of NMR brown adipose tissue and its UCP1 differ from those in mice and whether the lack of thermal insulation causes extreme changes in NMR body temperature upon cold exposure. Through longitudinal molecular, thermal, metabolic, and behavioral measurements, we found that NMRs initiated non-shivering thermogenesis and elevated body temperature but could not sustain it due to excessive heat loss and limits to substrate availability. Our results suggest that NMRs represent a unique thermoregulatory category that doesnt fit neatly into traditional classifications. In vitro and in vivo experiments showed that the NMR UCP1 is functional and can be activated and inhibited as expected for most other mammals. We further demonstrated that artificial insulation can partially restore thermoregulatory capabilities in NMRs. This study employs an advanced methodology to characterize the thermal biology of NMRs and helps resolve a long-standing controversy in the field. Significance StatementThis study provides critical insights into the thermal biology of naked mole-rats (NMRs), resolving long-standing controversies regarding their thermoregulatory strategies. We highlight the unique adaptations and limitations of NMR physiology by demonstrating that NMRs possess functional non-shivering thermogenesis and UCP1 but fail to maintain homeothermy due to excessive heat loss. Our findings suggest that artificial insulation can partially restore their thermoregulatory capabilities, offering a new perspective on the evolutionary and ecological significance of fur loss in NMRs. This research advances our understanding of mammalian thermal biology and presents an updated model for NMRs, bridging gaps between previous conflicting reports.

physiology↗

bioRxiv2025

A connectomic resource for neural cataloguing and circuit dissection of the larval zebrafish brain

We present a correlated light and electron microscopy (CLEM) dataset from a 7-day-old larval zebrafish, integrating confocal imaging of genetically labeled excitatory (vglut2a) and inhibitory (gad1b) neurons with nanometer-resolution serial section EM. The dataset spans the brain and anterior spinal cord, capturing >180,000 segmented soma, >40,000 molecularly annotated neurons, and 30 million synapses, most of which were classified as excitatory, inhibitory, or modulatory. To characterize the directional flow of activity across the brain, we leverage the synaptic and cell body annotations to compute region-wise input and output drive indices at single cell resolution. We illustrate the datasets utility by dissecting and validating circuits in three distinct systems: water flow direction encoding in the lateral line, recurrent excitation and contralateral inhibition in a hindbrain motion integrator, and functionally relevant targeted long-range projections from a tegmental excitatory nucleus, demonstrating that this resource enables rigorous hypothesis testing as well as exploratory-driven circuit analysis. The dataset is integrated into an open-access platform optimized to facilitate community reconstruction and discovery efforts throughout the larval zebrafish brain.

neuroscience↗

bioRxiv2025

A Cryptic Interfacial Pocket Uncovered in Full CRL4CRBN-IKZF3 Ubiquitylation Complex Enhances IMiD Efficacy

Immunomodulatory imide drugs (IMiDs) redirect the CUL4-RBX1-DDB1-CRBN (CRL4CRBN) ligase to ubiquitylate and degrade disease-linked proteins, but the full picture of how IMiDs glue the neosubstrate within the whole CRL4CRBN complex remains unknown. We determined cryo-electron microscopy structures of eight approved or clinical IMiDs in full CRL4CRBN ubiquitylation complexes at resolutions up to 3.4 [A], and revealed how these structurally distinct IMiDs exploit the structural plasticity of CRL4CRBN to organize conformationally conserved and compact active ubiquitylation assemblies with the neosubstrate IKZF3. Four "next-generation" IMiDs were found to additionally engage a cryptic gluing-driven interfacial (GDI) pocket at the non-degron zinc finger 3 (ZF3) of IKZF3, which contributes to their enhanced efficacy and neosubstrate specificity. The identification of cryptic gluing-driven pockets formed only in full ubiquitylation complexes provides new structure-directed design opportunities for IMiDs to improve therapeutic efficacy.

biochemistry↗

bioRxiv2025

UniCell: Towards a Unified Solution for Cell Annotation, Nomenclature Harmonization, Atlas Construction in Single-Cell Transcriptomics

Standardizing cell type annotations across single-cell RNA-seq datasets remains a major challenge due to inconsistencies in nomenclature, variation in annotation granularity, and the presence of rare or previously unseen populations. We present UniCell, a hierarchical annotation framework that combines Cell Ontology structure with transcriptomic data for scalable, interpretable, and ontology-aware cell identity inference. UniCell leverages a multi-task architecture that jointly optimizes local and global classifiers, yielding coherent predictions across multiple levels of the ontology-defined hierarchy. When benchmarked across 20 human and mouse datasets, UniCell consistently outperformed state-of-the-art tools, including CellTypist, scANVI, OnClass, and SingleR, in annotation performance, and sensitivity to low-abundance populations. In disease settings, UniCell effectively identified previously unseen cell types through confidence-guided novelty detection. Applied to 45 human and 23 mouse tissue atlases, UniCell enabled cross-dataset and cross-species harmonization by embedding cells into a unified latent space aligned with Cell Ontology structure. Moreover, when used to supervise single-cell foundation models, UniCell substantially improved downstream annotation accuracy, rare cell detection, and hierarchical consistency. Together, these results establish UniCell as a generalizable framework that supports high-resolution annotation, nomenclature standardization, and atlas-level integration, providing a scalable and biologically grounded solution for single-cell transcriptomic analysis across diverse biological systems.

bioinformatics↗

bioRxiv2025

Unique Gluing Effect of ASXL1 K351 Monoubiquitination Stimulates PR-DUB-Mediated Nucleosomal H2AK119Ub Deubiquitination

Protein ubiquitination plays a critical role in regulating fundamental biological processes through both proteolytic and nonproteolytic mechanisms. While classically known for its role in targeting proteins for degradation, ubiquitination can also modulate enzymatic activity. However, all of the known mechanisms of such modulation involve spatially constrained mechanisms that take place near the enzyme-substrate interfaces. Here, we report an unprecedented regulatory paradigm mediated by ubiquitination that is located distal to the enzyme-substrate interface but can still activate the polycomb repressive deubiquitinase (PR-DUB) complex. Using cryo-electron microscopy, molecular dynamics (MD) simulations, hydrogen-deuterium exchange mass spectrometry (HDX-MS), and kinetics assays, we determined that the ASXL1 K351 monoubiquitination promotes PR-DUB-mediated nucleosomal H2AK119Ub deubiquitination by stabilizing the catalytic pocket of PR-DUB, causing significant increase in maximal catalytic velocity (Vmax) but almost no change in the substrate binding affinity (Km). Structural analysis revealed that the ubiquitin moiety at ASXL1 K351 bridges the BAP1 and ASXL1 subunits, acting as an intramolecular "glue" that constrains the conformational dynamics of these subunits without altering the nucleosome binding interface. Further MD and HDX-MS results demonstrated the monoubiquitination of ASXL1 K351 enables PR-DUB to effectively maintain a spatial arrangement of the catalytic conformation that closely aligns with substrate cleavage. This study expands our understanding of the mechanisms of ubiquitin function to include intramolecular fastening with a molecular gluing effect and provides a deeper mechanistic understanding of PR-DUB activation in nucleosomal H2AK119Ub removal.

biochemistry↗

bioRxiv2025

Spatial transcriptomics elucidates the central role of cerebral vasculatures in the progression of Japanese encephalitis

The cellular and molecular mechanisms underlying the neuropathology of Japanese encephalitis virus (JEV) remain obscure. Herein, we designed Stereo-seq chips to simultaneously capture the in situ transcriptomes of both the host and JEV, constructing a comprehensive spatiotemporal pathological landscape for Japanese encephalitis (JE). This study reveals the central role of the vascular system in JE pathogenesis, particularly the meninges, which displayed the strongest signal of inflammation and cell death in the JEV-infected brain. The activation of the Ackr1+ endothelial cells, disruption of the blood-brain barrier, the migration of JEV-infected monocytes, secretion of immune factors by the infected cells, and the occurrence of pyroptosis and necroptosis form a positive feedback loop, resulting in an increasing influx of immune cells and tissue damage. JEV infection leads to neurological impairments, which may be attributed to the interaction between viral proteins and host cellular factors such as Stat1, Stat3, Nfkb1, and Sp1. As the vascular system serves as a central receiver and amplifier of inflammatory signals, regulating inflammation within the vascular system is essential in mitigating JE progression.

immunology↗

bioRxiv2025

Neutralizing antibodies elicited by Nipah virus G-head nanoparticle target diverse sites and inhibit receptor binding and fusion

Nipah virus (NiV) and Hendra virus (HeV), two highly pathogenic Henipaviruses (HNVs), pose a significant public health threat. The attachment glycoprotein (G) plays a crucial role in viral attachment and entry, making it an attractive target for vaccine and therapeutic antibody development. However, the antigenic landscape and neutralization sensitivity of the G protein remain poorly defined. Here, we systematically characterize 27 monoclonal antibodies (mAbs) elicited by NiV G head (GH) nanoparticle-immunized mice. Among these, 25 mAbs exhibit neutralizing activity against two major NiV strains, NiV-Malaysia and NiV-Bangladesh, with five mAbs also cross-inhibiting HeV infection. Although all mAbs target GH, they inhibit viral infection through distinct mechanisms, either by blocking receptor ephrin-B2 engagement with the G protein or by preventing viral membrane fusion. Notably, mAbs from two distinct groups conferred complete protection to hamsters against lethal NiV-Malaysia infection. Structural analysis of NiV GH in complex with four representative Fabs reveals four non-overlapping epitopes, including two novel antigenic sites and one shared protective epitope identified across species. Our study provides a comprehensive map of the antigenic determinants of NiV GH, elucidates the relationship between epitopes, neutralization mechanisms, and protection, and offers new insights and opportunities for antibody-based therapies and vaccine development. HighlightsMost mAbs induced in G head nanoparticle-immunized mice are potent neutralizers MAbs recognizing the G head block receptor binding or viral fusion Structural analysis shows mAbs target four distinct epitopes on G head protein Receptor competition site on G head is a shared protective antigenic site in human and multiple animals

immunology↗

bioRxiv2025

Infectome Analysis of Small Mammals in Southern China Reveals Ecological Associations and Emerging Threats from Diverse Pathogens

Small mammals harbor a diverse array of zoonotic pathogens. To date, however, metagenomic surveys of these species have primarily focused on viral diversity, with limited attention paid to bacteria and eukaryotic pathogens. Additionally, the ecological determinants of pathogen diversity within these mammals have not been systematically examined. Herein, we employed a metatranscriptomics approach to survey the pathogen infectome of 2,408 individual samples, representing lung, liver, and gut tissues from 858 animals collected throughout Guangdong province, China, with a study design that accounted for host species, tissue, season, and geographic location. We identified 76 pathogen species, including 29 RNA viruses, 12 DNA viruses, 5 bacteria, and 30 eukaryotic pathogens, 33 of which were newly discovered. Tissue distribution analysis revealed distinct organotropisms, suggesting varied transmission routes, while host distribution analysis showed that each animal carried an average of one pathogen, with 10 pathogens widely distributed among mammalian orders. Our characterization of the geographic and seasonal patterns revealed that pathogen richness was primarily influenced by region, host species, and season, while pathogen composition was largely shaped by host genetic distance. Collectively, these data provide the first comprehensive insight into the dynamics of the pathogen infectome in these key mammalian disease reservoirs, highlighting the major factors driving pathogen diversity and transmission.

microbiology↗