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Methods: metagenomic sequencing was performed on nasal secretion samples from fungal ball cavity, affected and contralateral middle nasal meatus in 30 MSFB patients, and from the healthy middle meatus in 30 controls. Taxonomic and functional profiling used Kraken2, Bracken, and HUMAnN2. Results: bacteria dominated the fungal ball cavity microbiome, with Haemophilus influenzae as a key biomarker. Negative association with Aspergillus flavus and enriched metabolic pathways supported H. influenzae survival and immune modulation.",{"@graph":69,"@context":122},[70,84,105],{"@type":71,"itemListElement":72},"BreadcrumbList",[73,77,79,82],{"item":74,"name":75,"@type":76,"position":8},"https://docshare.wps.com","Home","ListItem",{"item":78,"name":9,"@type":76,"position":14},"https://docshare.wps.com/document/",{"item":80,"name":40,"@type":76,"position":81},"https://docshare.wps.com/document/research-report/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/haemophilus-influenzae-dominance-in-fungal-ball-microbiome-revealed-through-multi-niche-metagenomic-sequencing/457721/",{"url":83,"name":65,"@type":85,"image":86,"author":91,"headline":65,"publisher":94,"fileFormat":97,"inLanguage":63,"description":67,"dateModified":98,"datePublished":99,"encodingFormat":97,"isAccessibleForFree":100,"interactionStatistic":101},"DigitalDocument",{"url":87,"@type":88,"width":89,"height":90},"https://docshare.wps.com/thumbnails/haemophilus-influenzae-dominance-in-fungal-ball-microbiome-revealed-through-multi-niche-metagenomic-sequencing/457721.png","ImageObject",300,407,{"name":92,"@type":93},"jay","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-10-06","2026-09-30",true,{"@type":102,"interactionType":103,"userInteractionCount":19},"InteractionCounter",{"@type":104},"ViewAction",{"@type":106,"mainEntity":107},"FAQPage",[108,114,118],{"name":109,"@type":110,"acceptedAnswer":111},"What was the main objective of this study?","Question",{"text":112,"@type":113},"The study used metagenomic sequencing to characterize the sinonasal microbiome in unilateral maxillary sinus fungal ball patients, emphasizing bacterial–fungal interactions and functional pathways related to pathogenesis.","Answer",{"name":115,"@type":110,"acceptedAnswer":116},"How were samples collected for metagenomic sequencing?",{"text":117,"@type":113},"For MSFB patients, nasal secretion samples were taken from three anatomical sites: fungal ball cavity, affected middle nasal meatus, and contralateral unaffected middle nasal meatus. Controls provided samples from the healthy middle nasal meatus.",{"name":119,"@type":110,"acceptedAnswer":120},"What key microbiome finding was observed in the fungal ball cavity group?",{"text":121,"@type":113},"Bacteria dominated the fungal ball cavity microbiome (98.53%), and Haemophilus influenzae was identified as a key biomarker. 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BMC Microbiology (2026) 26:15 [https://doi.org/10.1186/s12866-025-04546-8](https://doi.org/10.1186/s12866-025-04546-8)  \nBMC Microbiology  \nRESEARCH Open Access  \nHaemophilus influenzae dominance in fungal  ball microbiome revealed through multi-niche metagenomic sequencing  \nTing Yang 1,2,3, Yi Wang 1,2,3, Yuan Zhang 1,2,3, Chengyao Liu 1,2, Yun Zeng 1,2,3, Pengyu Shi 1,2,3, Jie Zhou 1,2,3, Yunchuan Li 1,2* and Hongzheng Wei 1,2,3*  \nAbstract  \nObjective This study employed metagenomic sequencing to characterize the sinonasal microbiome in patients with unilateral maxillary sinus fungal ball (MSFB), with specific emphasis on bacterial-fungal interactions and functional pathways implicated in fungal ball pathogenesis.  \nMethods The study enrolled 30 MSFB patients and 30 healthy controls. Nasal secretion samples were obtained from three anatomical sites in MSFB cases: fungal ball cavity (FC), affected middle nasal meatus (AM), and contralateral unaffected middle nasal meatus (UM) . And in the control group, samples were obtained from the healthy middle nasal meatus (HM) . Metagenomic sequencing of microbial DNA was performed using the Il lumina Novaseq platform. Taxonomic and functional analyses were conducted using Kraken2, Bracken, and HUMAnN2 .  \nResults Bacteria dominated the microbiome in the FC group (98 . 53%), with Haemophilus influenzae identified as a key biomarker (LDA score >5) . A negative correlation between H. influenzae and Aspergillus flavus was observed in the FC group (r = -0 .46, P = 0 . 013) . Functional pathways enriched in the FC group included amino acid biosynthesis (map00290), lipopolysaccharide biosynthesis (map00540), and fatty acid biosynthesis (map00061), supporting H. influenzae survival and immune modulation. FC microbiota showed reduced diversity and distinct composition compared to other groups (PERMANOVA, P \u003C 0. 001) . No significant differences were found in the composition of the microbiota between the bilateral middle nasal meatus groups of MSFB.  \nConclusion This study highlights H. influenzae as a critical bacterial biomarker in MSFB. The inverse relationship between H. influenzae and A. flavus may suggest competitive or immune-mediated interactions. These findings advance understanding of non-invasive fungal sinusitis. Future validation in larger fungal ball cohorts or invasive fungal sinusitis is warranted.  \nKeywords Metagenomic, Fungal sinusitis, Haemophil us, Aspergillus  \n*Correspondence: Yunchuan Li [1964liyunchuan@163.com](1964liyunchuan@163.com)[ ](1964liyunchuan@163.com)Hongzheng Wei [whzpeking@163.com](whzpeking@163.com)  \n1Department of Otolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing, China  \n2Beijing Laboratory of Allergic Diseases and Beijing Key Laboratory of Nasal Diseases, Beijing Institute of Otolaryngology, Beijing, China 3Department of Allergy, Beijing Tongren Hospital, Capital Medical University, Beijing, China  \n© The Author(s) 2025. Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit [http://creati](h","cbCairvXt1UkDMk7","https://ap.wps.com/l/cbCairvXt1UkDMk7","pdf",2721698,"English","# Abstract\n## Objective\n## Methods\n## Results\n## Conclusion\n# Introduction","[{\"question\":\"What was the main objective of this study?\",\"answer\":\"The study used metagenomic sequencing to characterize the sinonasal microbiome in unilateral maxillary sinus fungal ball patients, emphasizing bacterial–fungal interactions and functional pathways related to pathogenesis.\"},{\"question\":\"How were samples collected for metagenomic sequencing?\",\"answer\":\"For MSFB patients, nasal secretion samples were taken from three anatomical sites: fungal ball cavity, affected middle nasal meatus, and contralateral unaffected middle nasal meatus. Controls provided samples from the healthy middle nasal meatus.\"},{\"question\":\"What key microbiome finding was observed in the fungal ball cavity group?\",\"answer\":\"Bacteria dominated the fungal ball cavity microbiome (98.53%), and Haemophilus influenzae was identified as a key biomarker. A negative correlation between H. influenzae and Aspergillus flavus was also reported.\"}]","Haemophilus influenzae dominance in fungal ball microbiome revealed through multi-niche metagenomic sequencing | PDF",1790750234,23]