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<title>bioRxiv Subject Collection: Microbiology</title>
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This feed contains articles for bioRxiv Subject Collection "Microbiology"
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<link>https://www.biorxiv.org</link>
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<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.30.755482v1?rss=1">
<title>
<![CDATA[
Piezophilic activity dominates the Puerto Rico Trench deep subseafloor biosphere 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.30.755482v1?rss=1
</link>
<description><![CDATA[
Hydrostatic pressure is a defining feature of the deep ocean, yet how it structures microbial activity across the subseafloor biosphere is poorly understood. Here, we assess pressure effects on anabolic activity in seafloor and subseafloor communities across a margin-to-hadal-to-abyssal transect of the Puerto Rico Trench region (493-8,385 m water depth). Community-level responses to pressure reverse with burial, revealing a previously unrecognized shift toward piezophily in resident subseafloor communities down to 20 meters below seafloor (mbsf). In pressure-incubation experiments, atmospheric pressure yielded higher proportions of active cells in surficial sediments, whereas in situ pressure yielded higher proportions in the subseafloor. Pressure also structured active-community composition; in trench-axis sediments extending to 8.35 mbsf, JS1/Atribacterota populations belonging to geographically widespread hadal lineages exhibited a physiological response to in situ pressure, indicative of subseafloor piezophily. These findings suggest that hydrostatic pressure acts as an ecological filter after burial, favoring piezophilic traits that may support microbial persistence in the deep subseafloor biosphere.
]]></description>
<dc:creator><![CDATA[ Desmarais, M., Breusing, C., Finch, A. H., Baumas, C. M. J., Orsi, W. D., Plominsky, A. M., Morono, Y., Wehrmann, L. M., Graham, D., Pockalny, R. A., Spivack, A. J., Smith, D. C., Saucedo, B., Soto, S. R., Sostre-Cortes, J., Dekas, A. E., Bowman, J. S., D'Hondt, S., Bartlett, D. H. ]]></dc:creator>
<dc:date>2026-10-01</dc:date>
<dc:identifier>doi:10.64898/2026.09.30.755482</dc:identifier>
<dc:title><![CDATA[Piezophilic activity dominates the Puerto Rico Trench deep subseafloor biosphere]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.30.755081v1?rss=1">
<title>
<![CDATA[
RecBCD Prevents Bacterial Autoimmunity by Clearing Self-DNA Ends Produced During Replication Termination 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.30.755081v1?rss=1
</link>
<description><![CDATA[
Viral-induced DNA damage triggers both eukaryotic and prokaryotic innate immunity. Here, we report that self-DNA ends generated routinely during normal termination of bacterial replication suffice to trigger the P2-encoded OLD antiphage system. Normally, these physiological triggers are cleared by the dual helicase-nuclease activities of the conserved RecBCD enzyme, which acts as an immune safeguard analogous to mammalian TREX1 exonuclease and plant DBR1 debranching enzyme. However, when this clearance fails, the transient self-DNA ends persist and are recognized by ATP-bound OLD dimers via a positively charged channel across their ATPase subunits. ATP hydrolysis then activates OLD to cleave tRNAs, causing cytotoxicity. Single-cell microscopy reveals that dozens of OLD molecules associate with kilobases of adjacent DNA, amplifying the sparse physiological DNA end signal. Our findings reveal how an endogenous byproduct poses a constant autoimmune threat in bacteria, highlighting a universal molecular conflict across kingdoms and the convergent solutions evolved to resolve it.
]]></description>
<dc:creator><![CDATA[ Govande, A. A., Ramsey, B., Unlu, I., Amundsen, S. K., Nagy, T. A., Hassan, A., Smiley, M., Trautman, C., Whiteley, A. T., Smith, G. R., Kim, S., Lim, H. C. ]]></dc:creator>
<dc:date>2026-10-01</dc:date>
<dc:identifier>doi:10.64898/2026.09.30.755081</dc:identifier>
<dc:title><![CDATA[RecBCD Prevents Bacterial Autoimmunity by Clearing Self-DNA Ends Produced During Replication Termination]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.30.755304v1?rss=1">
<title>
<![CDATA[
Human-targeted drugs extensively reshape regulatory networks controlling virulence and antibiotic resistance in Salmonella Typhimurium 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.30.755304v1?rss=1
</link>
<description><![CDATA[
Human-targeted drugs are increasingly recognized to affect bacterial physiology, yet their impact beyond growth inhibition, and the underlying molecular mechanisms, remain poorly understood. Here, we systematically mapped how chemically diverse host-associated compounds influence virulence and intrinsic antibiotic resistance in Salmonella enterica serovar Typhimurium. Using a high-throughput reporter approach, we quantified transcriptional response of 26 key stress promoters in Salmonella against 2,415 compounds, including FDA-approved drugs, food-derived molecules, and human metabolites. We found that non-antibiotic compounds induce transcriptional responses distinct from those elicited by antibiotics, uncovering a regulatory shift from general stress responses to compound-specific regulation driven by ligand-responsive MarR- and TetR-family regulators. In particular, more than 200 non-antibiotic drugs modulate virulence or intrinsic resistance pathways through the central regulators SlyA or RamR, respectively. By integrating our large-scale dataset with machine learning, we further defined chemical features associated with ramA transcriptional activation in Salmonella, and predicted ~200 additional candidate inducers of intrinsic antibiotic resistance in this pathogen. Together, our results uncovered a large and previously unrecognized chemical landscape that reshapes bacterial stress responses through ligand-responsive regulatory mechanisms, with implications for how host-associated compounds influence pathogen virulence and antibiotic resistance.
]]></description>
<dc:creator><![CDATA[ Amstalden, M. K., Olayo-Alarcon, R., Brenzinger, S., Debande, L., Zannoni, A., Fiore, E., Boudrioua, A., Rashidian, A., Kronenberger, T., Wagner, S., Sharma, C. M., Mueller, C. L., Brochado, A. R. ]]></dc:creator>
<dc:date>2026-10-01</dc:date>
<dc:identifier>doi:10.64898/2026.09.30.755304</dc:identifier>
<dc:title><![CDATA[Human-targeted drugs extensively reshape regulatory networks controlling virulence and antibiotic resistance in Salmonella Typhimurium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.29.755260v1?rss=1">
<title>
<![CDATA[
Evaluation of Human Norovirus Replication in Salivary Gland Cells 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.29.755260v1?rss=1
</link>
<description><![CDATA[
Human noroviruses (HuNoVs) are the most common cause of non-bacterial foodborne illnesses worldwide. HuNoVs are notoriously difficult to culture, and current cell culture systems, such as human intestinal enteroids have limitations in efficiency and scalability, which has limited their applications in food safety studies. Recently, it has been shown that transformed salivary gland cell lines (including NS-SV-TT-DC) allow for replication and passaging of norovirus GII.4. Herein, we examined whether we can independently observe viral replication in salivary gland cells and if other strains of human norovirus can replicate in these cells. For this purpose, we first evaluated the replication of several norovirus strains in NS-SV-TT-DC cells by either infecting them with vesicle-cloaked viruses or total viruses in stool filtrates. Low levels of viral replication was observed and vesicle isolation did not improve viral replication. Next, we screened for replication of 50 norovirus samples from 9 genotypes, and 14 isolates demonstrated over a two-fold viral replication. We observed that the initial viral load in the samples does not have a significant effect on replication efficiency. We were also able to passage several isolates. Subsequently, we examined whether interferon inhibitors such as Ruxolitinib, TPCA-1, and BX795 would improve viral replication and observed negligible effects. Finally we tested multiple supplements including MgCl2, bile salt, ceramide, and TAK-779. Most supplements had little or no effect on HuNoV replication in NS-SV-TT-DC cells. Bile salt and TAK-779 showed limited, genotype-specific benefits. Altogether these results indicate that salivary gland cells allow limited replication of certain HuNoV isolates.
]]></description>
<dc:creator><![CDATA[ Harlow, J., Briggs, S., Pothier, E., Pham, K., Nasheri, N. ]]></dc:creator>
<dc:date>2026-10-01</dc:date>
<dc:identifier>doi:10.64898/2026.09.29.755260</dc:identifier>
<dc:title><![CDATA[Evaluation of Human Norovirus Replication in Salivary Gland Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.29.755541v1?rss=1">
<title>
<![CDATA[
Non-coding RNAs drive transcriptional network rewiring underlying thermal dimorphism in a human pathogenic fungus 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.29.755541v1?rss=1
</link>
<description><![CDATA[
As fungal pathogens pose mounting global challenges, decoding their complex adaptive traits remains a paramount academic priority. While previous studies have provided foundational insights into protein-coding variations and orphan genes, fully understanding such traits requires exploring upstream non-coding regulatory networks. Here, we present a high-resolution, rRNA-depleted temporal transcriptomic framework across reciprocal dimorphic transitions to decode how non-coding network rewiring involving in fungal adaption. Integrating temporal profiling with comparative genomics reveals that early ATP-related metabolic reprogramming and melanin biosynthesis direct morphogenesis via species-specific motifs. Further temporal network modeling uncovers a functional stratification where long non-coding RNAs orchestrate early stress responses, whereas circular RNAs drive morphological conversion. Functional validation confirms that hub non-coding transcripts mediate stress adaptation, with cross-species perturbation in the closest non-pathogen species altering phenotypic outcomes. Together, these findings demonstrate how system-level non-coding dynamics underlie the evolutionary innovation of thermal dimorphism, establishing a robust paradigm for understanding specialized fungal pathogenicity and evolution adaption.
]]></description>
<dc:creator><![CDATA[ Hu, X., Wang, J., Yang, E. ]]></dc:creator>
<dc:date>2026-10-01</dc:date>
<dc:identifier>doi:10.64898/2026.09.29.755541</dc:identifier>
<dc:title><![CDATA[Non-coding RNAs drive transcriptional network rewiring underlying thermal dimorphism in a human pathogenic fungus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.30.755635v1?rss=1">
<title>
<![CDATA[
Regulatory and metabolic integration accompany plasmid domestication during adaptation to a plant-associated lifestyle 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.30.755635v1?rss=1
</link>
<description><![CDATA[
Plasmids are major drivers of evolutionary innovation and ecological adaptation in prokaryotes. Plasmids that confer beneficial functions on their host may become domesticated through long-term coevolution, resulting in stable vertical inheritance within a bacterial lineage. How this process contributes to the evolution of bacteria associated with eukaryotic hosts remains poorly understood. Here, we investigate the function of the two large domesticated plasmids, LPP1 and LPP2, in Pantoea agglomerans associated with wheat. We show that loss of LPP1 reduces bacterial fitness during wheat colonization and impairs the utilization of diverse carbon sources. Comparative transcriptomics further reveal that curing of either plasmid extensively remodels chromosomal gene expression under both laboratory and wheat-associated conditions, whereas genome-scale metabolic reconstruction predicts that many metabolic pathways are jointly encoded by plasmid and chromosome. This functional integration extends beyond plasmid-specific pathways, as plasmid curing disrupts the utilization of substrates metabolized exclusively by chromosomally-encoded enzymes. Our results indicate that LPP1 promotes P. agglomerans association with wheat primarily through its regulatory and metabolic integration with the chromosome rather than through the simple addition of accessory genes. We propose that LPP1, and likely LPP2, represent chromosome-like symbiosis plasmids whose domestication reflects long-term selection for plant-associated lifestyle in Pantoea. Our findings suggest that persistent selection imposed by symbiotic lifestyles drives plasmid domestication, progressively transforming mobile plasmids into chromosome-like replicons that are integral components of bacterial genomes.
]]></description>
<dc:creator><![CDATA[ Vichare, S., Mahn-Ripcke, K., Romero Picazo, D., Soluch, R., Huelter, N. F., Dagan, T. ]]></dc:creator>
<dc:date>2026-10-01</dc:date>
<dc:identifier>doi:10.64898/2026.09.30.755635</dc:identifier>
<dc:title><![CDATA[Regulatory and metabolic integration accompany plasmid domestication during adaptation to a plant-associated lifestyle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.30.755704v1?rss=1">
<title>
<![CDATA[
Sensitivity of Human Coronaviruses (HCoVs) to Heme Metabolism: Mechanisms Underlying Antiviral Effects 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.30.755704v1?rss=1
</link>
<description><![CDATA[
Heme metabolism regulates the availability of heme, an iron-containing molecule essential for oxygen transport, mitochondrial respiration, electron transfer, and enzymatic reactions. Increasing evidence implicates heme metabolism in regulating viral infection, potentially representing an important innate antiviral mechanism. Components of both heme biosynthesis and degradation pathways exhibit antiviral activity against different viruses; however, their contributions and conservation across human coronaviruses remain incompletely defined. We investigated the impact of modulating heme metabolism on infection by seasonal human coronaviruses HCoV-OC43 and HCoV-229E. Treatment with 5-aminolevulinic acid (5-ALA) produced cell line-dependent antiviral effects, with robust inhibition in H1299 cells and selective HCoV sensitivity in MRC-5 cells. These differences correlated with intracellular accumulation of protoporphyrin IX (PPIX), suggesting that cellular metabolic capacity influences antiviral efficacy. In contrast, components of the heme degradation pathway, including heme oxygenase-1 (HO-1), carbon monoxide, and biliverdin, did not exhibit antiviral effects against HCoV-OC43 or HCoV-229E. PPIX and synthetic porphyrin CoPPIX inhibited early infection through direct viral inactivation. Collectively, these findings indicate that antiviral effects of heme metabolism against seasonal human coronaviruses are primarily driven by direct inactivation by PPIX rather than cellular antiviral responses induced by the heme degradation pathway
]]></description>
<dc:creator><![CDATA[ Conohan, N. T., Hirasawa, K. ]]></dc:creator>
<dc:date>2026-10-01</dc:date>
<dc:identifier>doi:10.64898/2026.09.30.755704</dc:identifier>
<dc:title><![CDATA[Sensitivity of Human Coronaviruses (HCoVs) to Heme Metabolism: Mechanisms Underlying Antiviral Effects]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.10.01.755474v1?rss=1">
<title>
<![CDATA[
A defined rumen-native microbial consortium is associated with reduced culling risk across commercial dairy herds 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.10.01.755474v1?rss=1
</link>
<description><![CDATA[
Background: Microbiome research has linked microbial community structure to host phenotype, but translating microbiome-based discovery into defined interventions with reproducible long-term host effects remains challenging. We evaluated whether a defined four-member consortium of rumen native microorganisms identified through associations with host phenotype could influence herd retention. Cull outcomes (sold or died vs. retained in herd) were analyzed across 20 controlled commercial dairy trials encompassing more than 40,000 cattle housed across 110 experimental pens, with each trial conducted for at least 5 months. Treatment pens received a defined daily dose of Galaxis Frontier (GF, Native Microbials, San Diego, CA), containing Pichia kudriavzevii, Clostridium beijerinckii, Ruminococcus bovis, and Butyrivibrio fibrisolvens, incorporated into the ration; control and treatment groups were otherwise managed equivalently. Cow-level time to culling was modeled within each trial, accounting for pen-level clustering, and trial estimates were combined using random-effects meta-analysis. Results: Daily treatment with rumen native microbes was associated with a 9% lower hazard of culling across 20 trials (hazard ratio = 0.91, 95% confidence interval: 0.86-0.96; P < 0.001). Between trial heterogeneity was negligible, and treatment effects were not significantly moderated by breed or geographic region. Detailed health records from one trial were examined to further characterize health outcomes associated with GF; treatment was associated with fewer transition-related disorders, particularly ketosis and metritis, whereas mastitis and lameness were largely unaffected. These findings suggest that the health effects associated with GF may be more pronounced for disorders linked to physiological and metabolic adaptation during the transition period than for health events more broadly. Conclusion: Across diverse commercial dairy environments, treatment with a defined consortium of rumen native microbes was associated with improved dairy cow retention. The consistency of this association across independent trials provides large scale evidence that targeted application of native microorganisms can translate into reproducible, long-term outcomes for the host. Complementary health event data suggest that improved transition period health and metabolic stability may contribute to this effect. These findings support the potential for microbiome targeted interventions to influence productive longevity under commercial conditions.
]]></description>
<dc:creator><![CDATA[ Marotz, C., Jacobi, T., Lefler, J., Anderson, B., Zhelev, I., Katulski, S., Goeser, J., Embree, M. ]]></dc:creator>
<dc:date>2026-10-01</dc:date>
<dc:identifier>doi:10.64898/2026.10.01.755474</dc:identifier>
<dc:title><![CDATA[A defined rumen-native microbial consortium is associated with reduced culling risk across commercial dairy herds]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.30.755614v1?rss=1">
<title>
<![CDATA[
Isolation and characterisation of Lytic Jumbo Klebsiella phage BMPCR_005026 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.30.755614v1?rss=1
</link>
<description><![CDATA[
Background: Klebsiella pneumoniae is a key causative agent of multidrug resistant nosocomial infections. Bacteriophages offer a promising alternative treatment, but the diversity of Klebsiella species requires the characterisation of a broad range of phages, particularly the poorly characterised jumbo phages. Materials and Methods: Phage BMCPR_005026 was isolated from sewage against Klebsiella pneumoniae. Whole genome sequencing, transmission electron microscopy and host range analysis against a diverse set of 40 clinically relevant Klebsiella strains were used to characterise the phage. Results: The virion had a myovirus morphology and genomic analysis placed it within an undefined family in Caudoviricetes where it forms a new species within the genus Alcyoneusvirus. BMCPR_005026 could form plaques on 3 of the 40 Klebsiella strains tested (7.5%) and formed halos. Conclusion: BMCPR_005026 represents a new species of phage within the genus Alcyoneusvirus with activity against 3 Klebsiella KL-types.
]]></description>
<dc:creator><![CDATA[ Lloyd-Evans, I., Ford, M., Hazzard, J., Bartis, C., Patel, F., Roemer, N., Wright, K., Porter, K., Coleman, M., Galyov, E., Megremis, S., Clokie, M. R., Millard, A. D., Michniewski, S., Wilkinson, R. C. ]]></dc:creator>
<dc:date>2026-10-01</dc:date>
<dc:identifier>doi:10.64898/2026.09.30.755614</dc:identifier>
<dc:title><![CDATA[Isolation and characterisation of Lytic Jumbo Klebsiella phage BMPCR_005026]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.30.755688v1?rss=1">
<title>
<![CDATA[
The HSV-1 LAT enhancer regulates locus-specific H3.1 enrichment during early lytic infection 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.30.755688v1?rss=1
</link>
<description><![CDATA[
The Herpes Simplex Virus 1 (HSV-1) genome undergoes extensive chromatinization following entry into the nucleus, yet how viral regulatory elements influence this process during lytic infection remains poorly understood. The HSV-1 latency-associated transcript enhancer (LTE) is well established as a key element in the regulation of latency and reactivation, but its role during the productive infection has not been defined. Here, we identify a role for the LTE in regulating chromatin composition on the lytic HSV-1 genome. Deletion of a 310-bp region within the LTE selectively reduced enrichment of the canonical histone H3.1 at specific viral lytic genes, including ICP4 and ICP8, without altering H3.3 enrichment. This phenotype was not reproduced using a second LTE deletion that disrupted enhancer activity, indicating that regulation of H3.1 enrichment is separable from the function of the enhancer. LTE-dependent H3.1 enrichment persisted in the absence of viral DNA replication and was accompanied by reduced occupancy of the H3.1 histone chaperone CAF-1. However, depletion of CAF-1 did not substantially reduce H3.1 enrichment, indicating that additional mechanisms contribute to H3.1 association with the viral genome. Together, these findings reveal that the HSV-1 LTE regulates locus-specific chromatin composition during early lytic infection and identify a previously unrecognized connection between a viral enhancer and histone H3.1 enrichment on the incoming viral genome.
]]></description>
<dc:creator><![CDATA[ Manuel, K. A., Fraser, T. A., Shipley, M. A., Neumann, D. M. ]]></dc:creator>
<dc:date>2026-10-01</dc:date>
<dc:identifier>doi:10.64898/2026.09.30.755688</dc:identifier>
<dc:title><![CDATA[The HSV-1 LAT enhancer regulates locus-specific H3.1 enrichment during early lytic infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.30.750856v1?rss=1">
<title>
<![CDATA[
Giant bacteria in gut microbiomes 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.30.750856v1?rss=1
</link>
<description><![CDATA[
The advent of sequencing-based omics has transformed our understanding of the gut microbiome, enabling microbial communities to be characterised at unprecedented scale and resolution. However, these approaches provide limited information about cellular traits such as size and morphology. Giant bacteria remain largely overlooked in gut microbiome research, despite their extraordinary cell size, extreme polyploidy and unusual reproductive strategies. Epulopiscium, one of the largest known heterotrophic bacteria, has been studied primarily as an intestinal symbiont of surgeonfish, leaving its broader host range unresolved. Here we show, by a large-scale analysis of publicly available 16S ribosomal RNA amplicon samples, alongside locally collected human samples, that Epulopiscium spp. occur across diverse hosts, including humans and a broad diversity of animals. Full-length 16S rRNA gene phylogenies resolved multiple Epulopiscium-related lineages with distinct host compositions. Among human samples, Epulopiscium sequences were most abundant during the first year of life. In adults, abundance varied across disease states, increasing in inflammatory bowel disease and colorectal cancer while decreasing in neurological conditions compared with a healthy adult baseline. Similar patterns were also observed in mouse models. Our study broadens the known host range of Epulopiscium and establishes giant bacteria as an overlooked component of gut microbiome diversity. These findings highlight the need to combine sequence-based studies with classical microbiological techniques to identify organisms with unique traits, otherwise hidden by taxonomy or phylogeny.
]]></description>
<dc:creator><![CDATA[ Lyu, Y., Gruener, M., Klein, A., Zoccaratto, L., Daya, A., Chang, H.-D., Bizic, M., Ionescu, D. ]]></dc:creator>
<dc:date>2026-10-01</dc:date>
<dc:identifier>doi:10.64898/2026.09.30.750856</dc:identifier>
<dc:title><![CDATA[Giant bacteria in gut microbiomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.29.755495v1?rss=1">
<title>
<![CDATA[
The protein phosphatase 2A catalytic subunit Pph22 partners with Tap42 and Rrd2 to control viability and virulence in Cryptococcus neoformans 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.29.755495v1?rss=1
</link>
<description><![CDATA[
Protein phosphatase 2A regulates eukaryotic signaling, but the function of its associated regulators remains poorly understood in the human fungal pathogen Cryptococcus neoformans. Here, we used affinity purification coupled with mass spectrometry and identified 81 candidate proteins associated with the catalytic subunit Pph22, including the scaffold subunit Tpd3, the regulator Tap42, and the protein phosphatase methylesterase 1. Conditional-expression and meiotic spore analyses supported an essential role for TAP42 in viability. We identified CNAG_05505 as the Rrd2 ortholog and detected an interaction between Rrd2 and Pph22 in a yeast two-hybrid assay. Deletion of RRD2 impaired thermotolerance, altered responses to multiple stresses and antifungal agents, reduced melanin and capsule production, delayed sexual development, and inhibited titan cell formation. Mutants also displayed altered cell wall staining and reduced expression of genes involved in chitin synthesis and deacetylation. RRD2 deletion increased basal Hog1 phosphorylation, whereas PPH22 overexpression reduced it. Double-mutant analysis revealed phenotype-dependent genetic interactions between RRD2 and HOG1. Although RRD2 deletion did not significantly alter overall survival in the Drosophila infection assay, it markedly attenuated virulence in mice: all mutant-infected mice survived the 70-day observation period, and fungal burdens were reduced across multiple organs at 21 days post-infection. These findings distinguish the essential contribution of Tap42 from the broad roles of Rrd2 in stress adaptation, morphogenesis, cell surface organization, and mammalian virulence, extending the functional analysis of the cryptococcal protein phosphatase 2A regulatory network.
]]></description>
<dc:creator><![CDATA[ Bahn, Y.-S., Yu, S., Cha, H., Won, D., Yu, S.-R., Seo, R., Lee, K.-A., Lee, J.-S., Lee, W.-J. ]]></dc:creator>
<dc:date>2026-10-01</dc:date>
<dc:identifier>doi:10.64898/2026.09.29.755495</dc:identifier>
<dc:title><![CDATA[The protein phosphatase 2A catalytic subunit Pph22 partners with Tap42 and Rrd2 to control viability and virulence in Cryptococcus neoformans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.10.01.755914v1?rss=1">
<title>
<![CDATA[
Adaptive respiratory bypass restores oxidative metabolism in NADH dehydrogenase-deficient Escherichia coli 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.10.01.755914v1?rss=1
</link>
<description><![CDATA[
Nicotinamide adenine dinucleotide is central to the cellular redox network and continuously cycles between its oxidized (NAD) and reduced (NADH) forms. Membrane-bound NADH dehydrogenases are the primary NADH-oxidizing enzymes and principal entry point for reducing equivalents into the bacterial respiratory chain. While maintaining optimal balance of this redox cofactor is among the most fundamental biochemical constraints on living systems, how cells adapt when the primary NADH oxidation route is lost remains poorly understood. Here we show that Escherichia coli lacking both NADH dehydrogenases initially compensates through costly fermentative lactate secretion, potentially to maintain redox and energy homeostasis, but at the cost of impaired growth and oxidative metabolism. Using adaptive laboratory evolution across four independent lineages, we found that all evolved populations derepressed malate:quinone oxidoreductase by disrupting an ArcA-binding site. This gain-of-function mutation establishes an alternative pathway that funnels electrons directly into the quinone pool, restoring respiratory oxygen consumption to wild-type level. Multi-omics and genome-scale metabolic modeling reveal that this bypass drives broad reorganization of central carbon metabolism, reinstates tricarboxylic acid cycle flux, and shifts cells toward a higher, more oxidative aero-type. These findings uncover a previously unrecognized metabolic buffering capacity that enables bacteria to circumvent loss of canonical respiratory entry points.
]]></description>
<dc:creator><![CDATA[ Biswas, A., Patel, A., Goel, N., Khairnar, S. V., Anand, A. ]]></dc:creator>
<dc:date>2026-10-01</dc:date>
<dc:identifier>doi:10.64898/2026.10.01.755914</dc:identifier>
<dc:title><![CDATA[Adaptive respiratory bypass restores oxidative metabolism in NADH dehydrogenase-deficient Escherichia coli]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.29.755485v1?rss=1">
<title>
<![CDATA[
Antibody-recruiting molecules based on mimotope fusion peptides and a dimeric antibiotic 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.29.755485v1?rss=1
</link>
<description><![CDATA[
New therapeutic approaches are needed to treat infections due to multidrug resistant (MDR) Gram-negative bacteria. The use of antibody-recruiting molecules (ARMs) is a promising approach to address the MDR problem given their potential to engage both the adaptive and innate immune systems to treat bacterial infections. Here we present two new strategies to create ARMs. First, we show that a 9-mer antimicrobial peptide (K6) can be fused to a 11-mer mimotope peptide recognized by antibodies targeting the Vibrio cholerae O1 O-antigen to produce an ARM peptide (FZ5) that can render E. coli more sensitive to complement killing in the presence of antibodies that recognize this mimotope. FZ5 can drive killing by complement in the presence of rabbit antibodies raised against O1 serogroup V. cholerae as well as antibodies present in human sera elicited by immunization with the live attenuated cholera vaccine PanChol. We also describe a strategy that combines bacterial surface binding with hapten display through the use of homodimers of the antibiotic streptomycin (Str) and anti-Str antibodies elicited by Str-protein conjugate vaccines. When combined with dimeric Str derivatives and human complement, polyclonal anti-streptomycin antisera kills E. coli by targeting the Str hapten. Dimeric forms of Str also significantly enhanced killing of a Str-resistant P. aeruginosa clinical isolate dependent on low levels of human complement and polyclonal anti-Str antibody. Together these data suggest that anti-bacterial ARM molecules based on mimotope display through simple synthetic fusion peptides, or dimeric haptens have promise as new approaches for the treatment of MDR bacterial infections.
]]></description>
<dc:creator><![CDATA[ Zingl, F. G., Mekalanos, M. L., Leitner, D. R., Walsh, S. R., Waldor, M., Mekalanos, J. J. ]]></dc:creator>
<dc:date>2026-09-30</dc:date>
<dc:identifier>doi:10.64898/2026.09.29.755485</dc:identifier>
<dc:title><![CDATA[Antibody-recruiting molecules based on mimotope fusion peptides and a dimeric antibiotic]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.29.755075v1?rss=1">
<title>
<![CDATA[
Capsule types driving carbapenem-resistant Klebsiella pneumoniae in Pakistan are largely absent from the phage literature 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.29.755075v1?rss=1
</link>
<description><![CDATA[
Phage therapy is increasingly proposed for carbapenem-resistant Klebsiella pneumoniae, but most Klebsiella phages bind a single capsule type, so a phage is useful only where its capsule type circulates. We asked whether published phages cover the capsule types found in Pakistani patients. We typed 254 clinical K. pneumoniae genomes from Pakistan and curated the phage and depolymerase literature into 34 phage-capsule records covering 27 phages and 15 capsule types. Nearly half the cohort was unmatched: 121 of 254 isolates (47.6%), spanning 51 of 66 capsule types, belong to types for which no phage has been described, and 67 of those isolates carry a carbapenemase. Collapsing each BioProject to one isolate per type raises the unmatched fraction to 51.8%. To ask why, we typed 475 genomes sampled on identical criteria from India, Bangladesh and Nepal, China, Europe and the USA. Two capsule types separated sharply. KL81 accounted for 6.3% of South Asian genomes and none of 275 from China, Europe or the USA, and has no published phage; KL107 showed the reverse, 9.8% in those regions and absent from South Asia. Both survived collapsing (P = 0.0014 and P = 0.0002). Six further differences did not, and are attributable to clonal expansion within single studies. Phages are isolated against locally available hosts, so the capsule types entering the global phage collection track the regions doing the isolating. KL81 is a defined target for phage isolation in South Asia.
]]></description>
<dc:creator><![CDATA[ Ahmed, T., Shah, A. H. ]]></dc:creator>
<dc:date>2026-09-30</dc:date>
<dc:identifier>doi:10.64898/2026.09.29.755075</dc:identifier>
<dc:title><![CDATA[Capsule types driving carbapenem-resistant Klebsiella pneumoniae in Pakistan are largely absent from the phage literature]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.29.755301v1?rss=1">
<title>
<![CDATA[
Single-cell transcriptomics reveals life cycle and cell cycle gene expression dynamics during in vitro growth of two Leishmania species 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.29.755301v1?rss=1
</link>
<description><![CDATA[
Leishmania species are transmitted between mammals by sand flies, within which they undergo a complex developmental programme involving transitions between replicative and non-replicative life cycle forms. Recent work using single-cell transcriptomics has revealed the developmental pathways of Leishmania major in the sand fly, but it is unclear how faithfully such development is recapitulated during Leishmania culture in vitro. In addition, no work has revealed how gene expression changes across the cell cycle of replicative Leishmania forms. Using single-cell transcriptomics on unsynchronised cultures of L. major and L. mexicana, we addressed both questions. We show that in vitro L. mexicana promastigotes display the same two developmental endpoints seen in L. major cells recovered from sand flies, metacyclics and haptomonads. In contrast, L. major promastigote development in vitro does not predict the presence of leptomonads or haptomonads, although non-dividing late metacyclics form through an early metacyclic stage as parasites enter stationary phase. Using computational reconstruction of the cell cycle in procyclic promastigote forms of both Leishmania species identified hundreds of genes whose transcript levels vary as the cell cycle progresses, revealing core conserved processes between the two species and with the closely related Trypanosoma brucei. These genes allowed us to examine connections between life cycle and cell cycle progression, revealing that some developmental transitions are strongly linked to a cell cycle phase, whereas others occur asynchronously. This work provides a comprehensive atlas and marker gene list for the interlinked cell and life cycles of two laboratory-cultured Leishmania species.
]]></description>
<dc:creator><![CDATA[ Marques, C. A., Laidlaw, R. F., Warren, F., da Silva, G. L. A., Young, M., Crouch, K., Burchmore, R., Otto, T. D., Llewellyn, M., McCulloch, R., Briggs, E. M. ]]></dc:creator>
<dc:date>2026-09-30</dc:date>
<dc:identifier>doi:10.64898/2026.09.29.755301</dc:identifier>
<dc:title><![CDATA[Single-cell transcriptomics reveals life cycle and cell cycle gene expression dynamics during in vitro growth of two Leishmania species]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.29.755374v1?rss=1">
<title>
<![CDATA[
An endonuclease V protects Bacillus subtilis against a deaminase toxin and a uracil-containing phage 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.29.755374v1?rss=1
</link>
<description><![CDATA[
Bacteria deploy polymorphic toxins to inhibit competitors and rely on cognate immunity proteins to prevent self-intoxication. Immunity determinants act by binding and neutralizing incoming toxins, and in the rare cases where toxin-induced damage is also reversed, that activity resides within the immunity protein itself. Whether a conflict system can instead dedicate a separate gene to repairing the damage is unknown. Here we report that an operon in Bacillus subtilis encodes the toxin YwqJ, its cognate immunity protein YwqK, and EndoV an endonuclease that incises DNA at the lesion the toxin creates. YwqJ is a single-stranded DNA cytidine deaminase translocated by the type VIIb secretion system (T7SSb) to mediate contact-dependent interbacterial antagonism. Unexpectedly, EndoV exhibits a preference for uracil over the canonical hypoxanthine, processing genomic uracil in cells and protecting against YwqJ when immunity is absent. Because EndoV acts on the chemical lesion rather than the effector, its protection is indifferent to the source of deamination. EndoVBs accordingly restricts replication of PBS1, a bacteriophage whose genome naturally substitutes uracil for thymine. These findings establish DNA repair as a novel layer of defense within toxin-immunity loci and reveal a DNA repair mechanism that unifies interbacterial antagonism and antiviral immunity.
]]></description>
<dc:creator><![CDATA[ Oliver, A. J., Joseph, S., Singh, A., Riazuddin, A., Lopez, M., Gao, Y., Bates, D., de Moraes, M. H. ]]></dc:creator>
<dc:date>2026-09-30</dc:date>
<dc:identifier>doi:10.64898/2026.09.29.755374</dc:identifier>
<dc:title><![CDATA[An endonuclease V protects Bacillus subtilis against a deaminase toxin and a uracil-containing phage]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.29.755287v1?rss=1">
<title>
<![CDATA[
Redox oscillation frequency reshapes antibiotic resistance-gene expression through metabolic and ecological selection in a freshwater community 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.29.755287v1?rss=1
</link>
<description><![CDATA[
Aquatic environments can maintain and express antibiotic resistance genes even without antibiotic pressure, but the conditions that govern their activity remain poorly understood. Redox oscillations are pervasive in natural and engineered waters and strongly alter microbial metabolism, yet whether their frequency shapes resistance expression is unknown. We reanalyzed paired metagenomes and metatranscriptomes from a published chemostat experiment in which a sulfidic stream community was cycled between oxic and anoxic conditions at high, medium and low frequency (HF, MF, LF), with strain-resolved population genomics of the dominant Pseudomonas lineage. Resistance genes were broadly distributed across treatments, whereas transcription concentrated under fast cycling, reaching a median 168 TPM under HF against 38 under LF although no antibiotic was present, so genetic potential and expression diverged. Fast cycling induced ROS-generating flavoproteins, antioxidant defenses and transcription-coupled DNA repair, and Pseudomonas supplied 99% of resistance-gene transcripts under HF, chiefly through its intrinsic efflux systems. Within the dominant Pseudomonas genome, however, the four resistance genes carried no sequence variants under HF, where coverage was deepest, and no fixed difference from the reference in any sample. Genome-wide selection remained purifying throughout (pooled pN/pS 0.72 under HF against 0.12 under LF, both below 1), and the relaxation trended toward redox and central metabolism. Redox frequency therefore raises resistance expression by selecting a metabolically versatile, high-efflux lineage, while the resistance sequences themselves stay conserved. Resistance activity in aquatic systems responds to an abiotic driver, which argues for weighing expression alongside gene presence in environmental surveillance.
]]></description>
<dc:creator><![CDATA[ Manna, B., Keenum, I., Liu, Q., Singhal, N. ]]></dc:creator>
<dc:date>2026-09-30</dc:date>
<dc:identifier>doi:10.64898/2026.09.29.755287</dc:identifier>
<dc:title><![CDATA[Redox oscillation frequency reshapes antibiotic resistance-gene expression through metabolic and ecological selection in a freshwater community]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.29.755237v1?rss=1">
<title>
<![CDATA[
Virophage-like viruses replicate with a candidate phycodnavirus of C. parva 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.29.755237v1?rss=1
</link>
<description><![CDATA[
TThe discovery of virophages and polinton-like viruses (PLVs) sparked studies of their hyperparasitic relationships with their associated giant viruses and hosts. To date, every cultured virophage and PLV depends on an Imitervirales helper virus. The freshwater alga Chrysochromulina parva is infected by viruses in the Imitervirales and Algavirales, and recent work showed that the PLV, CpV-PLV Moe, replicates via co-infection with the Imitervirales host CpV-BQ3. The other known PLVs, CpV PLV Larry and CpV PLV Curly, were subsequently isolated alongside CpV-BQ1, a putative Algavirales virus, raising the possibility that they depend on an Algavirales helper. To determine the nature of Larry and Curlys hyperparasitic relationships, infection experiments were conducted with each PLV and either CpV-BQ3 or CpV-BQ1. These experiments demonstrated that that neither Larry nor Curly replicated when introduced to C. parva alone, nor during mixed infections that included the Imitervirales virus BQ3. In contrast, the Algavirales virus CpV-BQ1 supported replication of both PLVs, with each increasing roughly 1000-fold over the course of infection while CpV-BQ1 replication was simultaneously reduced up to 100-fold. Together, these findings provide the first culture based evidence that PLV replication can be supported by an Algavirales helper and demonstrate that C. parva supports at least two distinct hyperparasitic relationships spanning different viral orders within the class Megaviricetes.
]]></description>
<dc:creator><![CDATA[ Thomas, G. R., Inamoto, I., Short, S. ]]></dc:creator>
<dc:date>2026-09-30</dc:date>
<dc:identifier>doi:10.64898/2026.09.29.755237</dc:identifier>
<dc:title><![CDATA[Virophage-like viruses replicate with a candidate phycodnavirus of C. parva]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.29.755191v1?rss=1">
<title>
<![CDATA[
Growth phase-dependent priming of galactose metabolism in Escherichia coli 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.29.755191v1?rss=1
</link>
<description><![CDATA[
Bacteria routinely respond to changing environments, with vastly different with nutrient availability. When nutritional changes are predictable, bacteria can deploy mechanisms to anticipate them. In these cases, an organism's past environment can shape its response to its current environment, which is often referred to as a history-dependent behaviour (HDB). Using nutritional shift assays, we investigated how prior growth phase influences subsequent growth across different carbon sources. Here, we show evidence of a novel HDB regulating galactose metabolism in Escherichia coli, where previous growth to exponential phase leads to a growth advantage in a galactose environment, with no comparable effect observed in all other carbon environments tested. We demonstrate a role of the galactose repressors, GalR and GalS, in regulating this behaviour, as it is lost in strains deficient in either repressor. Further, we show how this HDB is generalizable across a collection of E. coli isolates, but not Salmonella - a closely related species with a similar natural history. Together, these findings build on the existing knowledge of HDBs governing carbohydrate metabolism by demonstrating how growth phase history can influence an organism's response to fluctuating nutritional environments.
]]></description>
<dc:creator><![CDATA[ Mayol, J., Fatima, R., Surrette, M. G., Hynes, A. P. ]]></dc:creator>
<dc:date>2026-09-30</dc:date>
<dc:identifier>doi:10.64898/2026.09.29.755191</dc:identifier>
<dc:title><![CDATA[Growth phase-dependent priming of galactose metabolism in Escherichia coli]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.29.755562v1?rss=1">
<title>
<![CDATA[
Characterization of central carbon metabolism of Acinetobacter sp. Tol 5 based on 13C-metabolic flux analysis 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.29.755562v1?rss=1
</link>
<description><![CDATA[
Acinetobacter sp. strain Tol 5 is an attractive microorganism because of its ability to utilize various aromatic compounds, including toluene. However, because Tol 5 is a non-model microorganism, the utilization of carbon sources in central carbon metabolism remains unclear. In this study, the activity of the central carbon metabolism was characterized using 13C-metabolic flux analysis (13C-MFA). Tol 5 was cultured in synthetic medium using ethanol or acetate as the sole carbon source. In addition, because toluene is degraded into pyruvate and acetyl-CoA via the toluene dioxygenase pathway, this situation was mimicked by culturing with ethanol and lactate as co-substrates. The 13C-MFA confirmed flux through the glyoxylate shunt, which bypasses part of the TCA cycle when either ethanol or acetate was used as the sole carbon source. In ethanol, the flux ratio of isocitrate dehydrogenase at the isocitrate branch point decreased markedly. Unlike acetate, the conversion of ethanol to acetyl-CoA is accompanied by NADH production. Therefore, the observed change in this flux ratio was influenced by the excess NADH generated. Under ethanol-lactate conditions, Tol 5 simultaneously utilized ethanol and lactate. The ethanol consumption rate decreased to less than half of that observed when ethanol was the sole carbon source. Most of the lactate uptake was converted to pyruvate and metabolized via the TCA cycle. The glyoxylate shunt flux was lower than that in ethanol. These metabolic characteristics provide valuable insights into the use of Tol 5 as a next-generation microbial cell factory.
]]></description>
<dc:creator><![CDATA[ Miyamoto, E., Imada, T., Teraki, H., Miyoshi, K., Niide, T., Inoue, S., Yoshimoto, S., Hori, K., Shimizu, H., Toya, Y. ]]></dc:creator>
<dc:date>2026-09-30</dc:date>
<dc:identifier>doi:10.64898/2026.09.29.755562</dc:identifier>
<dc:title><![CDATA[Characterization of central carbon metabolism of Acinetobacter sp. Tol 5 based on 13C-metabolic flux analysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.29.755536v1?rss=1">
<title>
<![CDATA[
Oysters from five continents harbour a vast array of previously unknown RNA viruses, including many putative viruses of vertebrates 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.29.755536v1?rss=1
</link>
<description><![CDATA[
Oysters are vital to global aquaculture and marine ecosystems, yet, their associated RNA viruses remain largely unexplored. Here, we analysed ~1,000 publicly available metatranscriptomic datasets from 13 oyster species across 14 countries and 5 continents to characterize the oyster RNA virome (ORV). We identified 1,515 distinct RNA viruses putatively representing 17 viral orders and 766 species-level viral operational taxonomic units (vOTUs), with ~62% being previously undocumented putative species. These viruses not only populate extant branches of RNA virus evolution, but also represent previously unrecognized families and genera, and include viruses that putatively infect bacteria, fungi, protists, plants, invertebrates, and vertebrates. Based on sequence similarity, we found many calicivirus-like, orthoreovirus-like, astrovirus-like, and picornavirus-like viruses that do not infect invertebrates or microbes; thus, oysters accumulate diverse viruses from the surrounding water, and can potentially be interrogated as reporters of viruses circulating in the environment.
]]></description>
<dc:creator><![CDATA[ Zhong, K. X., Chan, A. M., Suttle, C. A. ]]></dc:creator>
<dc:date>2026-09-30</dc:date>
<dc:identifier>doi:10.64898/2026.09.29.755536</dc:identifier>
<dc:title><![CDATA[Oysters from five continents harbour a vast array of previously unknown RNA viruses, including many putative viruses of vertebrates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.29.755473v1?rss=1">
<title>
<![CDATA[
Leveraging genomes and metagenome-assembled genomes unveils novel diversity and transmission between public transport and clinical settings in Acinetobacter 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.29.755473v1?rss=1
</link>
<description><![CDATA[
The molecular epidemiology of Acinetobacter, which is almost exclusively culture-dependent, has centered mainly on clinical isolates of A. baumannii. Yet, this overlooks sources not amenable to microbial culture and many non-A. baumannii species. Here, by analysing metagenomic projects from public transport systems worldwide, we show that metagenome-assembled genomes (MAGs) are highly relevant for assessing the unrecognized diversity of Acinetobacter. We recovered 839 Acinetobacter MAGs, with 269 being high-quality MAGs. The high-quality MAGs were across more than 20 well-defined Acinetobacter species; however, five species (A. nosocomilalis, A. ursingii, A. radioresistens, A. schindleri, and A. variabilis) had the most MAGs. Pangenome and phylogenetic diversity analyses of 685 genomes plus the 269 high-quality MAGs from those 5 species showed that MAGs are an important source of both lineage and gene diversity. Also, FST analyses and genome phylogenies show that for some species there is no genetic differentiation between MAGs and clinical isolates, implying transmission between hospitals and public transport settings. In addition to finding the antibiotic resistance genes (ARGs) in MAGs, we implemented an ad hoc strategy (leveraging the use of genomes) to recover considerably more ARGs. PCoA and PERMANOVA analyses show that, whereas in A. nosocomialis and A. ursingii, the ARG profiles of MAGs and clinical genomes are different, in A. radioresistens and A. schindleri, the ARG profiles are not different. Finally, some clinically important ARGs, such as sul1 and sul2, aadA, and blaOXA23, were found in all or some of the 5 species. Our study shows how combining MAGs and regular genomes is a powerful strategy for better appraising the genomic diversity (including the resistome) of Acinetobacter. On a broader level, our study showcases the importance of MAGs for genomic surveillance of man-made environments with clear public health implications.
]]></description>
<dc:creator><![CDATA[ Lopez-Sanchez, R., Aguilar-Vera, A., Castillo-Ramirez, S. ]]></dc:creator>
<dc:date>2026-09-30</dc:date>
<dc:identifier>doi:10.64898/2026.09.29.755473</dc:identifier>
<dc:title><![CDATA[Leveraging genomes and metagenome-assembled genomes unveils novel diversity and transmission between public transport and clinical settings in Acinetobacter]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.29.755216v1?rss=1">
<title>
<![CDATA[
Functional Divergence in Gut Clostridia from Hadza Hunter-Gatherers Reveals Diet-Adapted Metabolic Machinery That Shapes Gut Ecology 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.29.755216v1?rss=1
</link>
<description><![CDATA[
Lifestyle changes associated with industrialization, including shifts in diet, are linked to extensive remodeling of the human gut microbiome. Investigating the gut microbiomes of traditional populations living non-industrialized lifestyles, such as the Hadza hunter-gatherers of Tanzania, therefore offers insight into bacterial taxa and functions depleted in industrialized microbiomes. Despite pronounced compositional differences that correspond to lifestyle, many commensal gut species are shared between industrialized and non-industrialized populations, raising a fundamental question of whether and how their functional repertoires diverge across contrasting gut environments. Here, by integrating strain-resolved metagenomics with culturomics and in vivo models, we identify phylogenetic and functional divergence within prevalent gut species shared between Hadza and industrialized populations, particularly among members of the Clostridia class. Dorea longicatena, prevalent across diverse human populations independent of lifestyle, is a poorly characterized member of Clostridia. Using strains of this D. longicatena isolated from either Hadza or industrialized human stool samples we uncover metabolic pathways selectively enriched in Hadza strains, including machinery for utilizing plant-derived pectic polysaccharides. We further demonstrate that this Hadza-enriched polysaccharide-utilization machinery can be leveraged with its cognate dietary substrate to promote D. longicatena colonization within a model industrialized microbiota, reshaping intestinal and portal-vein metabolite profiles. Together, our findings reveal substantial functional divergence within gut bacterial species shared across lifestyles, providing a framework for identifying diet-responsive strains and pathways that may complement ecological and metabolic functions underrepresented in industrialized microbiotas.
]]></description>
<dc:creator><![CDATA[ Takeuchi, T., Reddy, A., Robinson, E. K., Kotaka, K., Higginbottom, S. K., Sonnenburg, E. D., Sonnenburg, J. L. ]]></dc:creator>
<dc:date>2026-09-30</dc:date>
<dc:identifier>doi:10.64898/2026.09.29.755216</dc:identifier>
<dc:title><![CDATA[Functional Divergence in Gut Clostridia from Hadza Hunter-Gatherers Reveals Diet-Adapted Metabolic Machinery That Shapes Gut Ecology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.24.752487v1?rss=1">
<title>
<![CDATA[
The global evolutionary dynamics of natural capsule switching in a human-restricted bacterial pathogen 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.24.752487v1?rss=1
</link>
<description><![CDATA[
Exchange of genetic loci encoding extracellular capsular polysaccharides in bacteria can change capsular antigens or serotype, which, in turn, can lead to evasion of vaccines targeting the capsule. However, the evolutionary dynamics and drivers of bacterial capsule switching remain poorly understood. Here, we investigate the evolutionary dynamics and correlates of capsule switching in bacteria, using the human-restricted bacterial pathogen Streptococcus pneumoniae as an example, using 10,558 genetically and geographically diverse whole-genome sequences. Our phylogenomic and ancestral state reconstruction analysis revealed that the propensity for capsule switching varies by capsule type and genetic background. We found capsule switches only between ~4% of all possible capsule-type pairs, varying by specific pairs and switching direction. We also found associations between capsule switching patterns and genetic and ecological factors, including nasopharyngeal carriage duration, capsule biosynthesis locus size, and capsule thickness. Furthermore, the genetic similarity of the replacing and replaced capsule loci was primarily associated with the frequency of switching between specific capsule type pairs. Although a few specific non-capsule genomic loci were associated with the propensity for capsule switching across distinct lineages, the collective effect of the genetic background appears to influence capsule switching more strongly. These findings shed light on how bacterial capsule types and lineages undergo capsule switching influenced by genetic and ecological factors, informing our understanding of the emergence of novel capsule-genotype combinations that may impact the design of capsule-targeting glycoconjugate vaccines.
]]></description>
<dc:creator><![CDATA[ Chaguza, C., York, A., Lo, S. W., Kalizang'oma, A., Gladstone, R. A., Kwambana-Adams, B. A., Cave, R., Global Pneumococcal Sequencing (GPS) Consortium,, Klugman, K. P., Andam, C. P., Heyderman, R. S., McGee, L., Rosch, J. W., Croucher, N. J., Bentley, S. D., Weinberger, D. M. ]]></dc:creator>
<dc:date>2026-09-30</dc:date>
<dc:identifier>doi:10.64898/2026.09.24.752487</dc:identifier>
<dc:title><![CDATA[The global evolutionary dynamics of natural capsule switching in a human-restricted bacterial pathogen]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.26.754612v1?rss=1">
<title>
<![CDATA[
Biogenic flavonoid capping converts a cytotoxic Carica papaya fraction into a selective, cross-serotype Dengue entry inhibitor 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.26.754612v1?rss=1
</link>
<description><![CDATA[
Plant-derived flavonoids show measurable anti-dengue activity in cell culture, but their translational value is limited by a narrow therapeutic window: the concentrations that inhibit the virus approach or exceed those that are cytotoxic. Whether nanoparticle formulation can resolve this constraint, rather than simply add potency, remains untested for a chemically defined fraction. Here we show that biogenic silver nanoparticle (AgNP) formation using a flavonoid-enriched fraction of Carica papaya inverts an unusable selectivity profile into a viable one. The unformulated fraction was cytotoxic below the concentrations required for antiviral activity: its 50% cytotoxic concentration (CC50 = 134.9 ug/mL) lay below its 50% effective concentration against dengue virus serotype 2 (DENV-2; EC50 = 254.4 ug/mL), giving a Selectivity Index (SI) of 0.53. Using the same flavonoids as sole reducing and capping agents produced AgNPs (Z-average 128.4 nm; PDI 0.232; zeta potential -28.4 mV) that moved both parameters simultaneously. CC50 rose approximately 8.6-fold to 1,165.74 ug/mL while EC50 fell approximately 8.8-fold to 29.05 ug/mL, raising the SI to 40.12, an approximately 75-fold shift. Time-of-addition analysis localised the effect to the extracellular phase: inhibition was significant under pre-treatment and co-treatment but not after viral adsorption, identifying the AgNPs as entry inhibitors rather than replication inhibitors. Consistent with a serotype-independent physical mechanism, AgNP treatment at 30 ug/mL reduced viral RNA across all four serotypes, using inocula standardised against WHO-traceable NAAT reference reagents. These findings identify capping chemistry, rather than silver content alone, as a determinant of the therapeutic window in phytosynthesised nanoantivirals.
]]></description>
<dc:creator><![CDATA[ Dhandapani, M. K., Bharath, R. R., Palanisamy, G., Venkatasubbu, G. D., Bhatt, T. ]]></dc:creator>
<dc:date>2026-09-30</dc:date>
<dc:identifier>doi:10.64898/2026.09.26.754612</dc:identifier>
<dc:title><![CDATA[Biogenic flavonoid capping converts a cytotoxic Carica papaya fraction into a selective, cross-serotype Dengue entry inhibitor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.28.754943v1?rss=1">
<title>
<![CDATA[
Development of lyophilized faecal inoculation capsules for use in koala rehabilitation and conservation 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.28.754943v1?rss=1
</link>
<description><![CDATA[
As a specialist herbivore, koalas rely on their gut microbiomes to help digest their toxic and fibrous diet of Eucalyptus leaves. Without these critical microbes, koalas may not be able to obtain the nutrients and energy they need to survive. As such, a large proportion of koalas that undergo rehabilitation for chlamydiosis develop gut dysbiosis from the antibiotic treatment and are either euthanized or die. Here we aimed to modify previously developed fresh faecal inoculation capsules for an extended shelf-life such that they could be applied in a clinical setting to prevent or treat gut dysbiosis. Using 16S rRNA gene amplicon sequencing we demonstrated that air-drying faecal material leads to an overgrowth of facultative anaerobes, whereas lyophilised material retains a similar microbial composition to fresh material. Initial survival of koala faecal microbes as assessed by live/dead staining combined with microscopy was high after lyophilisation regardless of which excipient was used, except for 20% glycerol that resulted in ~15% lower survival than other treatments. Lyophilised fine particles extracted from koala faeces and packaged into acid-resistant capsules maintained their original microbial composition and had high microbial survival over a year, regardless of the excipient used. Dry-fill, single-layer capsules maintained integrity after 10 hrs in synthetic koala stomach acid. These capsules may be useful in adapting the gut microbiomes of koalas to novel diets e.g. during translocations. However, attempts to apply the capsules in the clinical setting were unsuccessful due to unanticipated difficulties in administrating the capsules to sick koalas.
]]></description>
<dc:creator><![CDATA[ Blyton, M. D. J., Dierens, L., Lacour, M., Hugenholtz, P. ]]></dc:creator>
<dc:date>2026-09-30</dc:date>
<dc:identifier>doi:10.64898/2026.09.28.754943</dc:identifier>
<dc:title><![CDATA[Development of lyophilized faecal inoculation capsules for use in koala rehabilitation and conservation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.28.755224v1?rss=1">
<title>
<![CDATA[
De novo minibinders targeting RH5 achieve nanomolar inhibition of blood-stage malaria replication 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.28.755224v1?rss=1
</link>
<description><![CDATA[
Malaria kills {approx}600,000 people annually despite recently deployed vaccines. The parasite's blood-stage invasion of erythrocytes, driven by the essential, non-redundant interaction between Plasmodium falciparum RH5 and erythrocyte basigin, remains a leading therapeutic target. Here, we designed de novo mini-proteins (minibinders) to occlude this interface. From {approx}1,500 computationally generated designs, we synthesized 18 candidates ranging from 9.3 to 16.9 kDa which were easily expressed in E. coli as soluble, well-folded proteins. Of these, 7 minibinders blocked blood-stage replication of P. falciparum asexual stage parasites with greater potency than {approx}300nM IC50. Three potent candidate minibinders, mb-5, mb-7, and mb-21, were confirmed to bind RH5 via SPR and BLI and block the merozoite-to-ring transition in stage-synchronized parasite progression assays, with CryoEM of minibinder-RH5 complex confirming the predicted binding at RH5-basigin interface. Although pharmacological challenges remain, our data demonstrate that potent inhibitors of parasite growth, rivaling existing antimalarials, can be designed rather than discovered via extensive screening. Our data highlight the potential for artificial intelligence to change the development landscape of new antimalarial therapies.
]]></description>
<dc:creator><![CDATA[ Hsiao, J., Shamshoum, M., Chalamalasetty, A., Barrett, J. R., Almada-Monter, R., Tabornal, E., Wilks, E., Lee, Y.-Z., Beutler, N., Lukens, A. R., Kapoor, P., Chen, D., Ndihokubwayo, J., Williams, B. G., McHugh, K., Pulido, D., de Sousa, I. M., Komives, E., Noormahomed, E. V., Wilson, I. A., Wirth, D. F., Rogers, T., Bethel, N., Draper, S. J., Ward, A., Winzeler, E. A., Jinich, A. ]]></dc:creator>
<dc:date>2026-09-29</dc:date>
<dc:identifier>doi:10.64898/2026.09.28.755224</dc:identifier>
<dc:title><![CDATA[De novo minibinders targeting RH5 achieve nanomolar inhibition of blood-stage malaria replication]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.28.755240v1?rss=1">
<title>
<![CDATA[
Reverse genetics system for emerging tick-borne orthonairovirus highlights dispensable N-terminal region of Gn associated with replication in tick vectors 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.28.755240v1?rss=1
</link>
<description><![CDATA[
A growing number of emerging and re-emerging tick-borne orthonairoviruses belonging to the Sulina and Tamdy genogroups have recently been identified in association with febrile diseases in East Asia. Yezo virus (YEZV) is one such viruses and is genetically distinct from well-characterized Crimean-Congo hemorrhagic fever virus. Orthonairovirus glycoproteins (Gn and Gc) expressed from a single glycoprotein precursor (GPC) gene mediate interaction with host cells, yet the organization and functions of the YEZV glycoproteins remain largely undefined. To characterize YEZV glycoproteins in the context of infection, we developed a reverse genetics system that enables recovery of recombinant YEZV entirely from cloned cDNAs. The recombinant wild-type virus exhibited growth properties comparable to those of the parental isolate in vitro and maintained pathogenicity in vivo. Proteomic analysis of purified virus particles produced in mammalian cells showed that peptide coverage of GPC-derived products began at residue 69. We therefore used the reverse genetics system to examine the functional importance of the GPC subregion upstream of residue 69. A mutant with a deletion of GPC residues 28 to 68 (rYEZV GPC{Delta}28-68), which retained the predicted signal peptide, was successfully recovered, suggesting that this subregion is dispensable for producing infectious virus. rYEZV GPC{Delta}28-68 propagated at levels comparable to those of the wild-type virus in mammalian cells and exhibited similar pathogenicity in a mouse model. Interestingly, the mutant reached lower viral titers in tick-derived ISE6 cells and in ticks in vivo, suggesting that this N-terminal subregion may contribute to virus replication in ticks. Together, these findings demonstrate the utility of the newly established reverse genetics system for dissecting the functions of the YEZV glycoproteins in mammalian and tick systems.
]]></description>
<dc:creator><![CDATA[ Mimura, Y., Hiono, T., Arakawa, H., Go, S., Shigeno, A., Ito, M., Fujii, E., Mizuma, K., Nakao, R., Orba, Y., Kaji, H., Matsuno, K. ]]></dc:creator>
<dc:date>2026-09-29</dc:date>
<dc:identifier>doi:10.64898/2026.09.28.755240</dc:identifier>
<dc:title><![CDATA[Reverse genetics system for emerging tick-borne orthonairovirus highlights dispensable N-terminal region of Gn associated with replication in tick vectors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.09.28.754992v1?rss=1">
<title>
<![CDATA[
In vitro antimicrobial resistance in clinical isolates of bacterial pathogens causing bovine respiratory disease: a systematic review and meta-analysis 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.09.28.754992v1?rss=1
</link>
<description><![CDATA[
Background: Bovine respiratory disease (BRD) is a major cause of antimicrobial use in cattle, raising concerns about antimicrobial resistance (AMR). This systematic review estimated pooled AMR proportions in BRD pathogens: Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni, and assessed temporal and geographic trends in resistance. Methods: Five antimicrobials commonly used for BRD treatment were evaluated: penicillin G (PEN), tetracycline (TET), florfenicol (FLO), tulathromycin (TUL), and enrofloxacin (ENR). Following PRISMA guidelines, 1,097 records were screened and 24 studies met inclusion criteria. Study quality and risk of bias were assessed, and random-effects meta-analyses estimated pooled proportions of resistant isolates by antimicrobial, continent and period. Results: A total of 13,949 bacterial isolates were analysed including P. multocida (5,615), M. haemolytica (5,585), and H. somni (2,749). TET resistance was highest across all pathogens (12-30%), followed by TUL (6-11%), PEN (4-12%), FLO (0.5-1.6%) and ENR (0.2-1.1%). TET resistance in M. haemolytica and P. multocida, and FLO resistance in P. multocida were significantly more frequent in North America than in Europe (p[&le;]0.05). TUL resistance increased significantly in P. multocida after 2013 (p=0.038), with similar non significant trends in M. haemolytica and H. somni. FLO resistance decreased significantly in H. somni after 2013 (p=0.013). Egger's test identified significant funnel plot asymmetry for ENR resistance in H. somni and PEN resistance in M. haemolytica, suggesting possible small study effects. Conclusion: These results offer insights into distribution of phenotypic resistance across pathogens and regions, informing BRD antimicrobial treatment guidelines and One-Health antimicrobial stewardship aimed at preserving the effectiveness of medically important antimicrobials.
]]></description>
<dc:creator><![CDATA[ Mahmood, Q., Rasmussen, P., Damborg, P. P., Hansen, K. B., Gregersen, C. B., Scahill, K. A., Carmo, L. P., Firth, C. L., Pardon, B., Stokstad, M., Anestad, L. M., Guardabassi, L., Conrady, B. ]]></dc:creator>
<dc:date>2026-09-29</dc:date>
<dc:identifier>doi:10.64898/2026.09.28.754992</dc:identifier>
<dc:title><![CDATA[In vitro antimicrobial resistance in clinical isolates of bacterial pathogens causing bovine respiratory disease: a systematic review and meta-analysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
</rdf:RDF>
