mSphere最新文献

{"title":"Imaging the lifecycle of <i>Microsporidia</i> sp. MB in <i>Anopheles coluzzii</i> from western Burkina Faso reveals octosporogony.","authors":"Ewan R S Parry, Roland Pevsner, Beth C Poulton, Deepak-Kumar Purusothaman, Abdelhakeem I Adam, Sare Issiaka, Thomas H Ant, Stephanie M Rainey, Etienne Bilgo, Abdoulaye Diabaté, Steven P Sinkins","doi":"10.1128/msphere.00851-24","DOIUrl":"10.1128/msphere.00851-24","url":null,"abstract":"<p><p>A newly discovered microsporidian, <i>Microsporidia</i> sp. MB (MB), was recently found to block <i>Plasmodium falciparum</i> transmission in <i>Anopheles arabiensis</i> mosquitoes from Kenya. Here, we describe the lifecycle of the first laboratory colony of <i>Anopheles coluzzii</i> with MB, originating from western Burkina Faso. The lifecycle of MB was explored using fluorescent <i>in situ</i> hybridization and confocal microscopy, facilitated by the development of optimized protocols to produce histological sections of whole adult, larval, and embryo tissues. As in <i>An. arabiensis</i>, transmission appears to be predominantly vertical, with MB highly localized to the ovaries across multiple lifecycle stages. MB was sparsely distributed within the majority of developing oocytes in the gravid female. After oviposition, in the majority of embryos, MB relocated to the developing gonad at the onset of tissue differentiation, suggesting a highly specialized adaptation to host tissues. Sporogony was identified for the first time in a proportion of developing oocytes and in embryos post-oviposition. Microsporidian spore characteristics were subsequently confirmed with electron microscopy. Identification of MB sporogony in eggs suggests there are alternative horizontal routes of transmission which could play an important role in developing MB as a malaria control strategy.</p><p><strong>Importance: </strong>Malaria in West Africa, caused by <i>Plasmodium falciparum</i> infection and spread by anopheline mosquitoes, is responsible for hundreds of thousands of deaths annually and resulted in over 120 million cases in 2022 . The transmission-blocking effect of <i>Microsporidia</i> sp. MB (MB) suggests its potential as an agent for combating the spread of malaria. Understanding the routes of transmission and their effect on MB in mosquito populations is crucial for its development as a control tool. The identification of MB spores reveals the potential for another avenue of transmission beyond the vertical transmission from female to offspring. Spores could also have the potential for alternative MB dissemination methods, alongside or instead of adult mosquito releases.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0085124"},"PeriodicalIF":3.7,"publicationDate":"2025-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12188704/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144119483","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genome content reorganization in the non-model ciliate <i>Chilodonella uncinata</i>: insights into nuclear architecture, DNA content, and chromosome fragmentation during macronuclear development.","authors":"Ragib Ahsan, Xyrus X Maurer-Alcalá, Laura A Katz","doi":"10.1128/msphere.00075-25","DOIUrl":"10.1128/msphere.00075-25","url":null,"abstract":"<p><p>Ciliates are a model lineage for studies of genome architecture given their unusual genome structures. All ciliates have both somatic macronuclei (MAC) and germline micronuclei (MIC), both of which develop from a zygotic nucleus following sex (i.e., conjugation). Nuclear developmental stages are not well documented among non-model ciliates, including <i>Chilodonella uncinata</i> (class Phyllopharyngea), the focus of our work. Here, we characterize nuclear architecture and genome dynamics in <i>C. uncinata</i> by combining 4',6-diamidino-2-phenylindole (DAPI) staining and fluorescence <i>in situ</i> hybridization (FISH) techniques with confocal microscopy. We developed a telomere probe for staining, which alongside DAPI allows for the identification of fragmented somatic chromosomes among the total DNA in the nuclei. We quantify both total DNA and telomere-bound signals from more than 250 nuclei sampled from 116 individual cells, and analyze changes in DNA content and nuclear architecture across <i>Chilodonella</i>'s nuclear life cycle. Specifically, we find that MAC developmental stages in the ciliate <i>C. uncinata</i> are different from those reported from other ciliate species. These data provide insights into nuclear dynamics during development and enrich our understanding of genome evolution in non-model ciliates.</p><p><strong>Importance: </strong>Ciliates are a clade of diverse single-celled eukaryotic microorganisms that contain at least one somatic macronucleus (MAC) and germline micronucleus (MIC) within each cell/organism. Ciliates rely on complex genome rearrangements to generate somatic genomes from a zygotic nucleus. However, the development of somatic nuclei has only been documented for a few model ciliate genera, including <i>Paramecium</i>, <i>Tetrahymena</i>, and <i>Oxytricha</i>. Here, we study the MAC developmental process in the non-model ciliate, <i>C. uncinata</i>. We analyze both total DNA and the generation of gene-sized somatic chromosomes using a laser scanning confocal microscope to describe <i>C. uncinata</i>'s nuclear life cycle. We show that DNA content changes dramatically during their life cycle and in a manner that differs from previous studies on model ciliates. Our study expands knowledge of genome dynamics in ciliates and among eukaryotes more broadly.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0007525"},"PeriodicalIF":3.7,"publicationDate":"2025-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12188737/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144006799","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genome mining identifies a diversity of natural product biosynthetic capacity in human respiratory <i>Corynebacterium</i> strains.","authors":"Ashley L Cunningham, Ilya Y Zhbannikov, Rachel Myers, Tommy H Tran, Wei Gao, Katherine P Lemon, Jhoanna N Aquino, Jillian H Hurst, Joon Won Yoon, Patrick C Seed, Matthew S Kelly","doi":"10.1128/msphere.00258-25","DOIUrl":"10.1128/msphere.00258-25","url":null,"abstract":"&lt;p&gt;&lt;p&gt;&lt;i&gt;Corynebacterium&lt;/i&gt; species, integral to the healthy human upper respiratory tract (URT) microbiota, remain underexplored in microbial genomics for their potential to promote respiratory health and exclude pathobionts. This genomic study investigated the diversity and capacity for natural product synthesis within these species, as indicated by their biosynthetic gene clusters (BGCs). We aimed to map and quantify the BGC diversity in a contemporary collection of &lt;i&gt;Corynebacterium&lt;/i&gt; strains, representative of their prevalence in the respiratory microbiota, and to elucidate intra- and interspecies variation in BGC content. The outcomes of this research could reveal key factors in maintaining the ecological balance of the upper respiratory tract and identify novel antimicrobial agents targeting respiratory pathobionts. Employing an &lt;i&gt;in silico&lt;/i&gt; approach, we analyzed the biosynthetic potential of respiratory strains of non-diphtheriae &lt;i&gt;Corynebacterium&lt;/i&gt; species and their reference genomes through genome sequencing and antiSMASH6 analysis. Among 161 genomes, we identified 672 BGCs, 495 of which were unique, including polyketide synthase, non-ribosomal peptide synthetase, ribosomally synthesized and post-translationally modified peptide, and siderophore families. To understand how this biosynthetic capacity compared to other respiratory bacteria, we then downloaded genomes from eight species that are associated with the URT and conducted BGC searches. We found that despite their compact genomes, &lt;i&gt;Corynebacterium&lt;/i&gt; species possess a multitude of predicted BGCs, exceeding the diversity of natural product BGCs identified in multiple other respiratory bacteria. This research lays the foundation for future functional genomics studies on the role of &lt;i&gt;Corynebacterium&lt;/i&gt; species in the respiratory microbiome and the discovery of novel therapeutics derived from this bacterial genus.IMPORTANCEBacterial secondary metabolites, produced by enzymes encoded by biosynthetic gene clusters, are ecologically important for bacterial communication and competition in nutrient-scarce environments and are a historically rich source of antibiotics and other medications. Human-associated &lt;i&gt;Corynebacterium&lt;/i&gt; species, abundant in the healthy upper respiratory tract, are understudied despite evidence of their roles in promoting human health and preventing pathobiont colonization. Through genome mining of a large collection of &lt;i&gt;Corynebacterium&lt;/i&gt; strains isolated from the human respiratory tract and publicly available genomes of other respiratory bacteria, our study suggests that &lt;i&gt;Corynebacterium&lt;/i&gt; species have a high biosynthetic capacity and are predicted to harbor a wide range of biosynthetic gene cluster families. These findings substantially expand current knowledge regarding the production of secondary metabolites by human-associated &lt;i&gt;Corynebacterium&lt;/i&gt; species. Our study also lays the foundations for understanding how &lt;i&gt;Corynebacterium","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0025825"},"PeriodicalIF":3.7,"publicationDate":"2025-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12188740/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144111455","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Sudapyridine (WX-081) inhibits <i>Mycobacterium tuberculosis</i> by targeting ATP synthase and upregulating host innate immunity.","authors":"Xinda Li, Xiaoyi Luo, Bin Wang, Lei Fu, Xi Chen, Yu Lu","doi":"10.1128/msphere.00149-25","DOIUrl":"10.1128/msphere.00149-25","url":null,"abstract":"<p><p>Drug-resistant tuberculosis (DR-TB) urgently requires safer, more accessible alternatives to bedaquiline (BDQ), which faces critical flaws like cardiotoxicity, high costs, and emerging resistance. WX-081, a promising BDQ alternative, has demonstrated superior anti-TB activity and improved safety in clinical studies. However, its mechanism of action remains unexplored, underscoring the need for further research to optimize its potential in advancing global TB elimination efforts. This study reveals WX-081's dual mechanisms: targeting <i>atpE</i> to disrupt ATP synthase and proton motive force via resistance screening, gene sequencing, and functional assays while enhancing host immunity through macrophage transcriptomics. Molecular docking confirmed <i>atpE</i> binding sites, and immune activation pathways (NF-κB/MAPK) were identified, positioning WX-081 as a potent, safe anti-DR-TB candidate despite unresolved mechanistic details.IMPORTANCEBedaquiline, a key drug for drug-resistant tuberculosis, is restricted by safety issues impacting its clinical utility. Its next-generation alternative, WX-081, has advanced to Phase III trials but lacks in-depth studies on its mechanism and host immune-modulatory effects, necessitating further research before broad clinical adoption.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0014925"},"PeriodicalIF":3.7,"publicationDate":"2025-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12188713/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144111456","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An extremely poor nutritional condition enables efficient white cell mating in <i>Candida albicans</i>.","authors":"Chaoran Zhong, Shuyun Guan, Ming Xu, Mingyang Ma, Chao Li, Li Tao, Guanghua Huang, Ming Guan","doi":"10.1128/msphere.00291-25","DOIUrl":"10.1128/msphere.00291-25","url":null,"abstract":"<p><p>Pathogenic fungi employ diverse strategies to undergo sexual reproduction. It was previously believed that the major human fungal pathogen <i>Candida albicans</i> must switch from the \"sterile\" white state to the mating-competent opaque state to mate. We recently reported that glucose depletion could induce <i>C. albicans</i> white cell mating at a moderate frequency. In this study, we demonstrate that white cells can undergo efficient mating when incubated on agar medium (water and agar, without additional nutritional components). Cell survival assays revealed that a subpopulation of white cells underwent cell death, releasing nutritional components that supported surviving cells in undergoing morphological changes and efficient mating. Inactivation of <i>CST20</i>, <i>MAC1</i>, <i>MCU1</i>, and <i>IDP2</i>-genes required for cell death and reactive oxygen species generation in <i>C. albicans</i>-resulted in a significant reduction in mating frequency under nutrient-depleted conditions. Taken together, our findings suggest that extremely poor nutritional conditions are able to promote epigenetic switch-independent mating in <i>C. albicans</i>.<b>IMPORTANCE</b>By demonstrating that white cells can mate under nutrient-depleted conditions, the research uncovers a novel mechanism of sexual reproduction in this pathogenic fungus. The findings suggest that cell death, through the release of nutritional components and the generation of reactive oxygen species, plays a crucial role in facilitating mating under nutrient-depleted conditions. This research not only provides novel insights into the reproductive strategies of <i>C. albicans</i> but also highlights potential avenues for exploring altruistic behaviors in other microorganisms. Understanding these mechanisms could have significant implications for the development of new therapeutic strategies to combat fungal infections, particularly in environments where nutrient limitations are common.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0029125"},"PeriodicalIF":3.7,"publicationDate":"2025-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12188724/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144226052","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Final reflections.","authors":"Michael J Imperiale","doi":"10.1128/msphere.00345-25","DOIUrl":"10.1128/msphere.00345-25","url":null,"abstract":"","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0034525"},"PeriodicalIF":3.7,"publicationDate":"2025-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12188736/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144226053","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"mSphere of Influence: Adapting new models for studying <i>Toxoplasma gondii</i> gut interactions.","authors":"Carlos J Ramírez-Flores","doi":"10.1128/msphere.00211-25","DOIUrl":"10.1128/msphere.00211-25","url":null,"abstract":"<p><p>Carlos J. Ramírez-Flores works in the field of parasitology, focusing on how <i>Toxoplasma gondii</i> crosses host barriers. In this mSphere of Influence article, he reflects on how three papers-by Shim, Kim, and Gregg-built on Dubey's foundational mouse work to reshape his view of the parasite's earliest intestinal events and to inspire his use of gut-on-chip platforms.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0021125"},"PeriodicalIF":3.7,"publicationDate":"2025-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12188729/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144234611","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Highlights of the 14th International <i>Bordetella</i> Symposium.","authors":"Kevin Munoz Navarrete, Kathryn M Edwards, Kingston H G Mills, Jana Kamanová, María Eugenia Rodriguez, Andrew Gorringe, Andrew Preston, Beate Kampmann, Monica C Gestal, Eric T Harvill, Purnima Dubey, Dimitri A Diavatopoulos, Seema Mattoo, Karen M Scanlon, Camille Locht, Peter Sebo","doi":"10.1128/msphere.00189-25","DOIUrl":"10.1128/msphere.00189-25","url":null,"abstract":"<p><p>Pertussis, or whooping cough, is a highly contagious and acute respiratory illness caused primarily by the gram-negative coccobacillus <i>Bordetella pertussis</i>. Despite near-universal vaccination, pertussis remains one of the least-controlled vaccine-preventable infectious diseases. Since 2023, pertussis incidence has been rising, and widespread pertussis outbreaks have resurged in many countries. In response to these emerging challenges, almost 300 experts from institutions across 24 countries convened at the 14th International <i>Bordetella</i> Symposium in Prague, Czech Republic, from 24 to 28 June 2024 to discuss pertussis epidemiology and research and strategies to mitigate the global pertussis burden. We present here the highlights of the symposium, comprising epidemiological and clinical aspects of <i>Bordetella</i> infections, results of clinical trials of pertussis vaccination in pregnant women and effectiveness of maternal vaccination in protecting newborn infants in Africa and Europe, the controlled human infection model (CHIM), and the latest insights into the biology, immunology, and pathogenesis of <i>B. pertussis</i> infection.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0018925"},"PeriodicalIF":3.7,"publicationDate":"2025-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12188742/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144079234","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The dynamics of <i>Cryptococcus neoformans</i> infection in <i>Galleria mellonella</i>.","authors":"Daniel F Q Smith, Aviv Bergman, Arturo Casadevall","doi":"10.1128/msphere.00190-25","DOIUrl":"10.1128/msphere.00190-25","url":null,"abstract":"<p><p><i>Galleria mellonella</i> has emerged as an important host for the study of fungal virulence, insect immune responses, and the evaluation of antifungal agents. In this study, we investigated the dynamics of fungal infections in <i>G. mellonella</i> using <i>Cryptococcus neoformans</i>, a human pathogenic fungus. Since the analysis of infection dynamics requires a fine temporal resolution of larval death, we employed a photographic time-lapse technique that allowed us to simultaneously measure death by proxy of larval melanization and absence of movement. Larval mortality occurred in two phases, early and late, which differed in their timing of melanization. Early phase deaths occurred with rapid whole-body onset of melanization, followed by sudden cessation of movement several hours later. Contrastingly, late phase deaths occurred with a gradual cessation of movement, followed by melanization, typically radiating from one location on the larva. The differences in mortality kinetics suggest differences in fungal pathogenesis, with one population succumbing early while the rest linger for later death. Subsequent analysis of mortality data using the inversion method revealed predictable deterministic dynamics but did not observe evidence of chaotic signatures. While this does not preclude the existence of chaos, it indicates that this <i>C. neoformans-G. mellonella</i> infection model may behave differently than bacterial-insect models.IMPORTANCEThe ability to predict the course of an infection is critical in anticipating disease progression and effectively treating patients. Similarly, the ability to make predictions about pathogenesis in laboratory infection models could further our understanding of pathogenesis and lead to new treatments. As fungal diseases are expected to rise, understanding the dynamics of fungal infections will be important to anticipate and mitigate future threats. Here, we developed a time-lapse method to visualize infections of <i>Galleria mellonella</i> larvae with the fungal pathogen <i>Cryptococcus neoformans</i>. This method provided insight into infection progression that is not apparent from standard survival measurement protocols, including the relationship between melanization and death. Further, it enabled us to explore the dynamics of disease progression in this system, which revealed deterministic dynamics without evidence of chaos, implying predictability in the outcome of cryptococcal infection in this moth.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0019025"},"PeriodicalIF":3.7,"publicationDate":"2025-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12188720/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144079236","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Significant associations between high-risk sexual behaviors and enterotypes of gut microbiome in HIV-negative men who have sex with men.","authors":"Kangjie Li, Xinjing Liu, Xiaohua Zhong, Haijiao Zeng, Tian Liu, Bing Lin, Pinyi Chen, Biao Xie, Xiaoni Zhong","doi":"10.1128/msphere.00232-25","DOIUrl":"https://doi.org/10.1128/msphere.00232-25","url":null,"abstract":"<p><p>Gut microbiome of men who have sex with men (MSM) exhibits distinctive characteristics compared with general populations. The dysbiosis of the gut microbiome in MSM is also associated with the onset and evolution of HIV infection. Enterotype is an important feature of the gut microbiome and remains unaffected by demographic factors. However, the enterotypes of gut microbiome in MSM are unclear. The associations between enterotypes and high-risk sexual behaviors in this population also remain to be elucidated. HIV-negative MSM were recruited in this study. Fecal samples of the participants were collected and subjected to 16S rRNA gene sequencing. Enterotype clusters were determined by Jensen-Shannon divergence based on genus-level relative abundance. Microbial function predictions were conducted by PICRUSt2 software. Univariate and multivariate logistic regression approaches were utilized to analyze the associations of enterotypes with sexual behaviors. A three-category random forest machine learning model was performed to further examine the correlation between abundant microbiome in each enterotype cluster and anal sex roles. Two enterotype clusters were identified in our data sets, primarily driven by genera <i>Phocaeicola</i> and <i>Segatella</i>. The alpha diversity was comparable between the two enterotype clusters. Microbial metabolic functions significantly differed, and multivariate logistic regression indicated a significant association between anal sex role and enterotype. The results of the three-category random forest model indicate that the dominant bacterial communities in gut enterotypes can effectively differentiate MSM who engage exclusively in receptive anal intercourse from those who engage in insertive or versatile anal intercourse (AUC: 0.6400, 0.6929, respectively). We identified two enterotype clusters of gut microbiome in HIV-negative MSM. Enterotypes of MSM were significantly associated with anal sex roles. These findings further highlight the close correlation between the gut microbiome and anal intercourse roles.</p><p><strong>Importance: </strong>Our study's discovery that gut microbiome enterotypes are significantly associated with anal sex roles in HIV-negative MSM opens a new frontier in understanding the complex interplay between microbiology and sexual health. This finding underscores the urgency of delving into the mechanistic connections between the gut microbiome, sexual behaviors, and HIV infection. By identifying modifiable factors influencing gut microbiome composition, we have paved the way for developing personalized preventive strategies that could disrupt the transmission dynamics of HIV within this high-risk population. This research contributes to the fundamental understanding of the gut microbiome's role in the sexual health of MSM, making it a pivotal advancement in the fields of gut microbiome research and sexual health.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0023225"},"PeriodicalIF":3.7,"publicationDate":"2025-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144485187","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}