Biology subjects

Engel, P.

Publications and source records attributed to Engel, P..

At least 19 recordsLinked to original sources

bioRxiv2026

Combination of enhanced gene activation and cell death causes lethal inflammatory pulmonary fibrosis

The current paradigm of inflammatory disease etiology is that it is caused by an aberrant increase in either gene activation or cell death. Studying the linear ubiquitin (linUb) chain assembly complex (LUBAC) was instrumental in proving that immune-receptor-dependent cell death can cause inflammatory disease. As LUBAC regulates both, gene activation and cell death, it can, however, also serve to evaluate the inflammatory-disease-initiating role of gene activation. Investigating mice with two different gene-activation-enhancing point mutations in LUBAC components showed that aberrantly increased gene activation in Hoil 1C458A/C458A mice did not cause pathology. In contrast, in HoipN101A/N101A mice it led to untoward cell death which, unexpectedly, was required for pathological inflammation, culminating in lethal lung fibrosis resembling human idiopathic pulmonary fibrosis (IPF). Hence, we here uncover a new etiology of inflammation, whereby aberrantly enhanced gene activation can be root cause of pathological, fibrogenic inflammation, yet intriguingly via inducing untoward cell death.

cell biology↗

bioRxiv2026

The diverse functions of viral microproteins

A large fraction of the uncharacterized proteome consists of microproteins. Viruses, rich in microproteins with potent activities, provide a unique system for exploring their functions and evolution. Here, we systematically annotate viral microproteins through a phenotype-first approach: construction of a >50,000-element small ORF library spanning the microprotein-coding potential of human-infecting viruses, followed by broad screens that enrich candidates for high-resolution profiling by Perturb-seq, mass spectrometry, AI-guided structure prediction, and mechanistic studies. This atlas identified >2,000 active microproteins that modulate host pathways through diverse mechanisms, including molecular mimicry, hijacking of key regulators, and altering subcellular localization. We further show the oncogenic potential of BNLF2b, an uncharacterized Epstein-Barr virus microprotein linked to nasopharyngeal carcinoma. Three principles emerge: viral microproteins are diverse and often multifunctional, show widespread convergence toward shared functions, and serve as reservoirs of evolutionary innovation. These findings elucidate how viral microproteins occupy a large functional landscape, establish them as versatile tools for host manipulation, and provide a framework for functional annotation of uncharacterized proteins, contributing toward predictive systems virology.

systems biology↗

bioRxiv2026

Species boundaries structure competitive interactions among honeybee gut bacteria

Bacterial species often harbor extensive strain-level diversity, raising the question of whether strains, rather than species, are the relevant ecological units. We quantified pairwise interactions among 12 strains from four prevalent Lactobacillus species of the honeybee gut microbiota using gnotobiotic bees. Microbiota-depleted bees were colonized with individual strains and all 66 pairwise combinations, with strain abundances measured by colony counts and strain-resolved amplicon sequencing. Negative interactions predominated, with most significant interactions involving mutual inhibition. Within-species interactions were stronger and more asymmetric than between-species interactions, producing less even community compositions. Across repeated cycles of colonization in microbiota-depleted bees, three of four within-species pairs lost one strain, whereas all four between-species pairs persisted. These results indicate that closely related strains experience greater niche overlap and stronger competition than strains from different species, supporting bacterial species as ecologically differentiated units while highlighting the importance of strain-specific traits in shaping community assembly and stability.

microbiology↗

bioRxiv2026

Temporal, spatial, and parasitic drivers of microbial variation in European honey bees

Although the roles of host-associated microbiomes in animal health are increasingly recognised, the factors influencing their variation remain understudied. The relatively simple microbiome of honey bees is a relevant system to address this gap. In particular, the relationship between variations in microbiome composition and the ectoparasite Varroa destructor, the main threat to honey bee health worldwide, is poorly established. In this study, we used metagenomic and statistical analyses of 1442 European honey bee colonies to investigate the relationships between the honey bee microbiome, temporality, location, V. destructor load, and behavioural response to its infestation by the host. While season, year, and location were identified as the main drivers of microbiome variation, V. destructor load emerged as a significant factor associated with microbiome variation. Notably, we identify several pathogens and opportunists that correlated positively with V. destructor load, while the core symbiont Bombilactobacillus correlated negatively. This is compatible with a shift in the microbiome toward dysbiosis, which may be driven by or promote V. destructor parasitism. By contrast, we found only limited evidence of an association between the microbiome and resistance behaviours of the host against this parasite. While the study cannot establish causal relationships, we present the largest metagenomic analysis of honey bee microbiomes to date, providing robust, generalisable evidence about the factors driving variation in the microbiome composition of this ecologically and economically important pollinator. These findings may serve as additional markers in selective breeding programs targeting V. destructor resistance, which could ultimately improve honey bee health.

microbiology↗

bioRxiv2026

Competition between co-localized gut symbionts underlies inter-individual variation in the honeybee gut microbiota

Microbes in the animal gut compete to colonize spatially restricted host niches. However, whether competition for space drives inter-individual variation among hosts, and which factors determine the outcome of such competition, remain poorly understood. Here, we investigated competitive interactions of the western honeybee gut symbiont Frischella perrara with other members of the bee gut microbiota. Shotgun metagenomics analysis of individual bees revealed that F. perrara is negatively correlated with a specific species of the genus Gilliamella. Co-colonization of microbiota-depleted bees with these two bacteria resulted in their competitive exclusion. The outcome of this competition depended on the relative number of bacteria each bee received and benefited Gilliamella when one of the two type VI secretion systems of F. perrara was mutated. Using fluorescently tagged strains, high-resolution microscopy, and gut region-specific quantification, we show that both bacteria localize to the same host niche in the ileum of mono-colonized bees, indicating competition in a spatially restricted host niche. Moreover, both microbes protected against infection, promoting bee health. This competition provides an explanatory mechanism underlying variation in the occurrence of F. perrara across honeybee colonies and highlights the importance of gut spatial structure and microbial competition in shaping microbiome composition and inter-individual variability.

microbiology↗

bioRxiv2026

3,500 years of sheeppox virus evolution inferred from archaeological and codicological genomes

Sheeppox virus (SPPV) is a major livestock pathogen causing economic hardship through reduced production and death of vulnerable sheep, with written descriptions of sheeppox-like disease recorded since antiquity. We report 21 novel ancient SPPV genomes spanning the Eurasian steppe Bronze Age ([~]1,700 BCE) to the Early Modern period in Western Europe, including multiple genomes obtained from medieval parchment. We estimate that major capripoxvirus lineages diverged [~]11,500-3,700 years ago, overlapping known translocations and bio-cultural developments in sheep. Our dataset supports SPPV diverging first within the lineage leading to goatpox virus and lumpy skin disease virus, and that known gene inactivation events within SPPV and goatpox virus occur in our earliest SPPV genomes. These findings reveal that the food security of Eurasian communities have been threatened by sheeppox for over 3,700 years, and provide new insights to the genomic evolution and potential host adaptation of sheeppox virus. TeaserAncient viral genomes from manuscripts and teeth illuminate the evolutionary history of sheeppox virus.

genetics↗

bioRxiv2025

Integrative taxonomy and distribution of Apis species in Malaysia

Southeast Asia is the centre of diversification for honey bees (genus Apis). Hence, accurate identification of honey bees found here is instrumental in paving the way for studying evolutionary relationships, biogeographic patterns, and management of these key pollinators. In this study, we characterized Apis species diversity across peninsular Malaysia and Borneo Malaysia using an integrative approach combining morphological traits and mitochondrial markers (CO1 and 16S rRNA). Field collections included both managed and wild colonies of A. mellifera, A. cerana, A. dorsata, A. florea, and A. andreniformis from peninsular Malaysia, as well as two additional cavity-nesting species of Borneo A. koschevnikovi and A. nuluensis. Species identification was performed based on morphological features, nesting ecology, and subsequently validated through sequencing. Samples collected from peninsular Malaysia showed congruence between field/morphological and molecular identification. In contrast, cavity-nesting bees from Borneo exhibited overlapping morphological characteristics leading to misidentification of species. These findings suggest the reliability of carrying out field-based identification of honey bees in peninsular Malaysia but requires molecular validation for Borneo region. The limitations of morphology-based interpretation for Borneo cavity-nesting bees highlights the need for more population-level and speciation studies on these species.

ecology↗

bioRxiv2025

Host and microbial factors influence bacterial colonization of the honey bee gut

The guts of many animals are colonized by host-specific microbes, yet the extent to which host filtering (host-derived constraints) shapes microbial colonization and host specificity remains poorly understood. Here, we used gnotobiotic honey bees (Apis mellifera) as a model system to systematically assess the colonization potential of a phylogenetically and ecologically diverse panel of 56 bacterial strains, spanning native symbionts, opportunistic bee-associated taxa, gut microbes from other bee species, and non-bee environmental isolates. Bacterial load and colonization frequency were quantified by strain-specific qPCR seven days post-inoculation, in monocolonization and in the presence of a synthetic community composed of native honeybee core bacteria. Bacterial load was highest for native strains and declined with increasing phylogenetic distance from native symbionts. Co-colonization with the synthetic community reduces load across all groups, but native strains were least affected. Across strains, completeness of KEGG metabolic pathways correlated with bacterial load in some ecological groups; however, metabolic capacity alone did not fully explain colonization patterns, either in monocolonization or under competitive conditions. A key finding was that in vitro sensitivity to antimicrobial peptide (AMPs; apidaecin, abaecin, defensins, hymenoptaecin) varied widely among strains and was highest in closely related bee-associated bacteria. Notably, even highly successful colonizers such as Gilliamella and Snodgrassella were AMP-sensitive. AMP sensitivity showed a negative correlation with bacterial load, but not with the frequency of host colonization. These findings suggest that AMPs modulate symbiont abundance rather than acting as strict barriers to colonization. Overall, our results reveal that host filtering in the bee gut is multifaceted, integrating immune-mediated barriers, microbial traits, and competitive interactions.

microbiology↗

bioRxiv2025

Gut microbiota variation across sympatric stingless bee species and honey bees in the Neotropics

Stingless bees (Meliponini) are ecologically and culturally important pollinators with a long tradition of human management in the Neotropics. Yet, little is known about how their gut microbiota vary across geographic regions or whether microbial exchange occurs with managed honey bees (Apis mellifera), which are often kept in close proximity. Using full-length 16S rRNA gene sequencing of individual bees sampled from 167 colonies, we characterized gut microbial community structure through a hierarchical, taxonomic and phylogenetic comparative framework, contrasting the microbiota of Melipona quadrifasciata and Melipona mondury with that of Apis mellifera across their shared geographic range in Brazil. The core microbiota of Melipona was dominated by Lactobacillus, Bifidobacterium, Apilactobacillus, Bombella, and Floricoccus, and showed inverse variation in relative abundance with lower-prevalence bacterial taxa. Although the core microbiota of the two stingless bee species overlapped only partially with that of Apis mellifera, they exhibited comparable alpha-diversity and beta-diversity dispersion, indicating broadly similar community assembly processes and dynamics. Nevertheless, we found that 6% of all amplicon sequence variants (ASVs) were shared between hosts, encompassing nearly all canonical honey bee "core" symbionts, indicating frequent spillover. Remarkably, several ASVs of Snodgrassella, a genus typically rare in stingless bees, reached high abundance in several M. quadrifasciata individuals and formed a deeply divergent clade ([~]96% 16S rRNA gene identity to S. alvi). These patterns are consistent with the hypothesis that human-mediated management practices, such as mixed apiaries and artificial feeding, create opportunities for microbial exchange between native and non-native bees. Together, our findings indicate that stingless bee gut microbiomes are compositionally stable yet ecologically permeable, shaped by both long-term host specificity and recent anthropogenic contact.

ecology↗

bioRxiv2025

Engineered symbiont biosensor maps micron-scale sugar gradients in the honeybee gut

The honeybee gut microbiota plays a key role in shaping host health and susceptibility to disease. Yet, the nutrient environment it experiences within the gut remains poorly characterized. In particular, little is known about the spatial distribution of nutrients across this community, as resolving such fine gradients in vivo has been technically challenging. Here, we engineer the native honeybee symbiont Snodgrassella alvi as a living biosensor to quantify the bioavailability of the dietary sugar arabinose within the gut. By expanding the genetic toolkit for S. alvi through chromosomal integration of high-burden genes and a suite of low-strength promoters, we achieve stable multi-gene expression without compromising host colonization. The resulting biosensor generates a specific, dose-dependent response to arabinose in situ, enabling visualization of sugar gradients across gut-associated bacterial biofilms at micron-scale resolution. Using this system, we show that diet-derived arabinose distribution is highly heterogeneous and can be influenced by the metabolic activity of co-colonizing Gilliamella species. These findings highlight how diet composition and microbial specialization generate fine-scale microenvironments within the gut. More broadly, this work further establishes S. alvi as a genetically tractable platform for in situ probing of microbial metabolism and nutrient distribution.

synthetic biology↗

bioRxiv2025

Priority effects determine community composition at the strain level in the honeybee gut microbiota

Gut microbial communities often differ at the strain level among individual hosts, but the mechanisms driving this variation remain poorly understood. One potential factor is priority effects, a process in which differences in the timing and order of microbial colonization influence subsequent community assembly ("first come, first served" dynamics). We hypothesize that such priority effects operate at the strain level within species, where closely related bacteria exhibit niche overlap, and that these dynamics can lead to community divergence even under similar environmental conditions. We tested these predictions, using the gut microbiota of honeybees, which harbor conserved microbial communities that differ in strain composition among individual bees. We sequentially colonized microbiota-depleted honeybees with two distinct microbial communities composed of the same twelve core microbiota species but different strains, ensuring that individuals shared species-level composition but differed at the strain level. We found that firstcomer strains consistently dominated the resulting communities, suggesting strong priority effects. Dropout experiments in which the firstcomer strain of a species was removed led to only partial increases in the colonization success of the conspecific latecomer, suggesting that both intra- and inter-species interactions contribute to priority effects. Our findings highlight the significant role of priority effects in strain-level community assembly and reveal their influence in shaping the specialized gut microbiota of honeybees, with important implications for the development of probiotic strategies in beekeeping. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=134 SRC="FIGDIR/small/689826v1_ufig1.gif" ALT="Figure 1"> View larger version (32K): org.highwire.dtl.DTLVardef@b0930eorg.highwire.dtl.DTLVardef@8687c5org.highwire.dtl.DTLVardef@3ba838org.highwire.dtl.DTLVardef@a10c3e_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology↗

bioRxiv2025

Real-time volatilomics reveals microbiota and pathogen fingerprints in the honey bee

Understanding the complex relationship between gut microbiota and their hosts often relies on invasive sampling techniques. Honey bees provide a tractable model for host-microbe studies. Here we establish single-bee volatilomics using secondary electrospray ionization (SESI-HRMS) to examine volatile organic compounds released to the air around an individual live honey bee. Specifically, we focused on primary gut microbiota metabolites present in gnotobiotic bees. Our findings reveal distinct volatilome profiles in honey bees that depend on their gut bacterial colonization state. We cross-validated our findings using an established metabolomics technique, LC-HRMS, to compare and contrast the metabolites detectable with each mass spectrometry-based method. Finally, we assessed the ability of SESI-HRMS to detect colonization with the bee pathogen S. marcescens. By comparing the volatile signature of this bacterium grown in liquid culture with that of infected honey bee headspace, we identified overlapping compounds, including butane-2,3-diol, that were absent in uninfected bees. SESI-HRMS volatilomics, paired with LC-HRMS, therefore has potential to identify non-invasive biomarkers of bee microbiome composition and infection at the level of individual insects. These biomarkers represent practical targets for the development of simple, field-ready diagnostic tools for monitoring pollinator health.

microbiology↗

bioRxiv2025

In vivo genome-wide CRISPRi-Seq in Streptococcus pneumoniae reveals host-adaptive pathways essential for meningitis and identifies new therapeutic targets

Streptococcus pneumoniae is an important human pathogen that causes invasive diseases and remains the leading cause of bacterial meningitis. However, the bacterial determinants required for survival within the central nervous system (CNS) are poorly understood. Here, we performed a genome-wide CRISPR interference screen (CRISPRi-Seq) in an in vivo zebrafish model to systematically map pneumococcal fitness determinants during meningitis. Using this approach, we investigated essential and non-essential genes contributing to the pathogenesis of pneumococcal meningitis. The screen identified 244 loci whose repression significantly reduced bacterial fitness in vivo, representing pathways involved in virulence, metabolism, cell-envelope biogenesis, translation, and stress adaptation. Comparative analysis with in vitro datasets showed that metabolic genes such as purA, proABC, thrC, glyA, and manLMN, as well as those involved in peptidoglycan and capsule biosynthesis (pbp3, cps2 operon) and oxidative stress response (nox, dpr), become selectively more essential in vivo. Functional validation confirmed that these pathways are critical for virulence, nutrient-dependent growth, and oxidative stress response during meningitis. Untargeted metabolomics of infected zebrafish corroborate several observed bacterial genetic sensitivities in the CNS with altered levels of key nutrients such as glucose and 4-aminobenzoic acid. Integration of the CRISPRi-Seq dataset with antibiotic-target annotations revealed both established and previously unrecognised vulnerabilities, including aminoacyl-tRNA synthetases such as leucyl-tRNA synthetase (LeuRS), whose inhibition by the benzoxaborole epetraborole improved host survival, highlighting its potential as a new therapeutic strategy for multidrug-resistant (MDR) S. pneumoniae. Together, these findings demonstrate the power of in vivo CRISPRi-Seq to define the pneumococcal essentialome under physiological conditions and reveal that metabolic adaptation, cell-envelope maintenance, and the oxidative stress response are central to bacterial survival in the CNS. This fitness map advances our understanding of pneumococcal adaptation in the CNS and identifies promising targets for developing therapies against S. pneumoniae.

microbiology↗

bioRxiv2025

Microbes release lower-value metabolites at higher rates

Microbial interactions are shaped by the exchange of metabolites, ranging from intermediates in central carbon metabolism to amino acids, vitamins and fermentation by-products. While specific metabolic processes such as overflow metabolism are well documented, a general principle explaining the chemical diversity of metabolite release and the variation in individual release rates remains elusive. Here, using a combination of computational and experimental approaches on seven different microbes, we show that release rates are negatively correlated with the value of metabolites, and that this relationship explains more variability in release rates than other frequently associated factors. By measuring metabolite release from 67 E. coli mutants lacking individual transport genes, we find that transporters both cause and counteract net metabolite release. These findings suggest that metabolite release constitutes a fitness cost that microbes have evolved to reduce, while revealing the underlying transport mechanism. This unifying explanation reshapes our understanding of how microbial interactions emerge and persist.

microbiology↗

bioRxiv2025

CaV1.2-dependent excitation-transcription coupling modulates nociception

Peripheral nociceptive sensory neurons integrate various noxious inputs, resulting in local depolarization that triggers the firing of action potentials and thus the sensation of pain. We recently reported that nociceptor depolarization itself initiates signaling by the calcium channel Cav1.2 causing acute hyperalgesia in vivo. However, whether this mechanism initiates excitation-transcription (E-T) coupling and thereby leads to long-lasting modulation of nociceptor activity remains poorly understood. Using high content imaging of dorsal root ganglion (DRG) neurons, we here found that depolarization of nociceptors induces phosphorylation of the transcription factor (TF) cAMP-response element binding protein (CREB), which was affected by inhibition of protein kinase A (PKA) and calcineurin, but not Ca2+/calmodulin-dependent protein kinases. Genetic deletion or pharmacological inhibition of Cav1.2 confirmed its role in calcium-dependent kinase signaling and CREB phosphorylation after depolarization. In line with this, pharmacological modulation of Cav1 channels affected the expression of a subset of depolarization-regulated immediate early genes known to orchestrate a broader transcriptional response. Indeed, RNA-Seq analysis of DRG neurons from mice with a tissue-specific deletion of Cav1.2 in nociceptive sensory neurons (SNS-Cacna1c-/- mice) revealed downregulation of multiple calcium and potassium channel subunits as well as proteins involved in synaptic vesicle release and cell adhesion. Furthermore, repetitive firing of action potentials and release of the neuropeptide CGRP was impaired in Cav1.2-deficient sensory neurons. SNS-Cacna1c-/- mice showed increased sensitivity to noxious heat and exacerbated inflammatory but not neuropathic pain. In conclusion, our data suggest a Cav1.2-dependent E-T coupling mechanism in nociceptors that counteracts nociception in vivo. HighlightsO_LIDepolarization induces Cav1.2- and PKA-dependent phosphorylation of the transcriptional regulator CREB in nociceptors C_LIO_LICav1.2 regulates the expression of depolarization-induced immediate early genes and induces reorganization of the signaling network in nociceptors C_LIO_LICav1.2-deficient nociceptors downregulate multiple ion channel subunits as well as proteins involved in synaptic vesicle release and cell adhesion C_LIO_LICav1.2-deficient nociceptors show impaired depolarization-induced PKA-II signaling, CREB phosphorylation, repetitive firing of action potentials, and release of the neuropeptide CGRP C_LIO_LINociceptor-specific Cav1.2-deficiency in mice increases the sensitivity to noxious heat and exacerbates inflammatory pain C_LI In briefIsensee et al. report that depolarization of nociceptors induces calcium influx through Cav1.2 channels, thereby initiating a signaling cascade that links neuronal excitation to transcriptional regulation of immediate early genes (E-T coupling). Nociceptor-specific knockout of Cav1.2 led to transcriptional changes in nociceptor-specific genes and was associated with impaired depolarization-induced signaling, action potential firing, and neuropeptide release. As the Cav1.2-deficient mice show increased noxious heat sensitivity and exacerbated inflammatory pain, these findings suggest that Cav1.2-dependent E-T coupling mechanism counteracts nociception in vivo.

neuroscience↗

bioRxiv2025

ReCycled: A Tool to Reset the Start of Circular Bacterial Chromosomes

Summary (two sentences)For many publicly available bacterial genomes the chromosome start is not set at the replication initiation protein. This is a burden for comparative genomics studies as the synteny between such genomes cannot readily be assessed. Here, we present ReCycled, a tool for identifying and resetting the start of circular bacterial chromosomes. Availability and implementationFreely available on GitHub at https://github.com/Freevini/ReCycled under the GPL-3.0 license. Runs on all tested GNU/Linux systems. ContactVincent Somerville, vincent@somerville.earth Supplementary InformationThe analyses, scripts and data to test the different pipelines are deposited online on zenodo (10.5281/zenodo.15170502).

bioinformatics↗

bioRxiv2025

Strain-level and phenotypic stability contrasts with plasmid and phage variability in water kefir communities

Microbial communities can change in response to top-down factors, such as phages, and bottom-up factors, such as nutrient availability. Previous studies have successfully investigated bacterial species-level dynamics, but diversity and interactions beyond the species-level is usually lacking. Traditional fermented foods, such as water kefir, provide ideal systems to study ecological and evolutionary dynamics beyond the species-level, as they are simple and trackable systems that are cultivated in non-sterile, nutrient-rich environments which foster microbial growth and invasion. Despite the central role of only a few lactic acid bacteria for fermentation, little is known about the genomic diversity and dynamics of these community members over time. Within the framework of a graduate course, 35 students propagated water kefir across several generations under different nutrient conditions and in different households to study microbial responses over time. We found that water kefir communities were generally stable at the species-level, with only rare bacterial species replaced over long timescales (more than 2 years). While we observed little strain-level diversity with few strain replacements over long timescales, closely related strains exhibited variation in accessory gene content, often encoded on plasmids, particularly those involved in ecologically meaningful functions such as sugar utilization pathways and phage defense systems. We hypothesise that these genomic variations could reflect the adaptations of strains to different sugars and phages. Consistent with this, we observed a diverse array of phages, many likely originating from the unique household environments. By documenting the genomic landscape of microbial species, strains, plasmids, and phages, this study advances our understanding of the diversity and dynamics of microbial communities in fermented foods. Furthermore, our course material is publicly available and offers a blueprint for bridging the gap between teaching and research, inspiring the next generation of scientists to unravel the complexities of microbial ecosystems. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=137 SRC="FIGDIR/small/640646v1_ufig1.gif" ALT="Figure 1"> View larger version (31K): org.highwire.dtl.DTLVardef@56db6borg.highwire.dtl.DTLVardef@5f6dc6org.highwire.dtl.DTLVardef@11fff14org.highwire.dtl.DTLVardef@1a37a9f_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology↗

bioRxiv2024

Symbiont loss and gain, rather than co-diversification shapes honeybee gut microbiota diversity and function

Studying gut microbiota evolution across animals is crucial for understanding symbiotic interactions but is hampered by the lack of high-resolution genomic data. Honeybees, with their specialized gut microbiota and well-known ecology, offer an ideal system to study this evolution. Using shotgun metagenomics on 200 worker bees from five honeybee species, we recover thousands of metagenome-assembled genomes and identify several novel bacterial species. While microbial communities were mostly host-specific, we found both specialists and generalists, even among closely related bacterial species, with notable variation between honeybee hosts. Some bacterial generalists emerged host-specific only at the strain level, suggesting recent host switches. While we found some signal of co-diversification between hosts and symbionts, this was not more than expected by chance and was much less pronounced than what has been observed for gut bacteria of hominids and small mammals. Instead, symbiont gains, losses, and replacements emerged as the predominant process for honeybees. This highly dynamic evolution of the specialized honey bee gut microbiota has led to taxonomic and functional differences across hosts, such as the ability to degrade pollen-derived pectin. Our results provide new insights into the evolutionary processes govern gut microbiota diversity across closely related hosts and uncover the functional potential of the previously underexplored gut microbiota of these important pollinators.

microbiology↗