Biofilm最新文献

{"title":"Engineering an electroactive bacterial cellulose-carbon nanotube composite membrane against Staphylococcus aureus","authors":"Daniel S. Levin ,&nbsp;Camila S. Cué Royo ,&nbsp;Denis Johnson ,&nbsp;Soumalya Ghosh ,&nbsp;Sricharani Rao Balmuri ,&nbsp;Huda Usman ,&nbsp;Shakira M. Martínez Vásquez ,&nbsp;David Kumar Yesudoss ,&nbsp;Abdoulaye Djire ,&nbsp;Mostafa Bedewy ,&nbsp;Tagbo H.R. Niepa","doi":"10.1016/j.bioflm.2025.100305","DOIUrl":"10.1016/j.bioflm.2025.100305","url":null,"abstract":"<div><div><em>Staphylococcus aureus</em> is the leading cause of skin infections in the U.S., and its rapid evolution and resistance to antibiotics create a barrier to effective treatment. In this study, we engineered a composite membrane with bacterial cellulose and carbon nanotubes (BC-CNT) as an electroactive dressing to rapidly eradicate vancomycin-intermediate <em>S. aureus</em>. Nonpathogenic <em>Komagataeibacter sucrofermentans</em> produced the BC membrane at an air-liquid interface. Then, carboxyl-functionalized multi-walled CNTs were integrated into decellularized BC to create stable and electrically conductive BC-CNT dressings. The electric potential and ionic flux across BC-CNT were modeled and standardized via chronoamperometry for experimental validation. We found that treatment with electroactive BC-CNT increases <em>S. aureus</em> sensitivity to vancomycin and prevents macro-scale biofilm formation. The bactericidal efficacy of the composite membrane is consistent with electrochemical stress caused by voltage mediated with BC-CNT. After a single hour of combinatorial electrical and drug treatment, biofilm-forming capacity was inhibited by nearly 92 %. These results advance applications of electrochemistry in medicine and create a new direction to overcome <em>S. aureus</em> infections on skin and soft tissues.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"10 ","pages":"Article 100305"},"PeriodicalIF":5.9,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144694323","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Farnesol emulsion for elimination of Pseudomonas aeruginosa biofilm in a 3D airway model of cystic fibrosis","authors":"Li Tan ,&nbsp;Khiry Sutton ,&nbsp;Sean V. Murphy ,&nbsp;Nicole Levi","doi":"10.1016/j.bioflm.2025.100317","DOIUrl":"10.1016/j.bioflm.2025.100317","url":null,"abstract":"<div><div>Cystic fibrosis (CF), a life-shortening genetic disease, is hallmarked by mucus obstruction, respiratory deficiency, and chronic bacterial infections. <em>Pseudomonas aeruginosa</em> is the most common virulent respiratory pathogen that is detrimental to the overall survival of CF patients. Here we evaluate the efficacy of farnesol emulsion, a broad-spectrum agent recently used to combat <em>P. aeruginosa</em> biofilm infections, for reducing <em>P. aeruginosa</em> infections in CF using a three-dimensional (3D) airway “organ tissue equivalent” (OTE) model. OTEs are fabricated using cells derived from human primary cells sourced from CF donors (CF-OTEs), which accurately recapitulate multiple key traits of human CF airways, including increased mucin accumulation and lower cilium beating frequency, compared to OTEs derived from normal donors (N-OTEs). The OTE model closely approximates the native CF condition to provide a platform where both mucoid and nonmucoid <em>P. aeruginosa</em> establish biofilms. Luminescence quantification and viable bacterial enumeration demonstrated that more <em>P. aeruginosa</em> biofilm mass developed upon CF-OTEs compared to non-CF (normal) OTEs. The capability to establish infection and biofilm formation, without acute tissue toxicity, allows for rapid discrimination of therapeutic efficacy in an accurate, human <em>in vitro</em> model. Farnesol emulsion disrupted <em>P. aeruginosa</em> biofilms <em>in situ</em> and also protected OTE lung cell viability. We propose that the 3D airway OTE infection model is a reliable preclinical tool for CF drug screening, with farnesol emulsion being a prospective drug candidate to treat <em>P. aeruginosa</em> biofilm infections in CF.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"10 ","pages":"Article 100317"},"PeriodicalIF":4.9,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145121175","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Combating biofilm formation and bacterial killing: N-acetylcysteine's efficacy against Pseudomonas aeruginosa in urinary catheters","authors":"Arthika Manoharan ,&nbsp;Greg Whiteley ,&nbsp;Rajesh Kuppusamy ,&nbsp;Slade Jensen ,&nbsp;Trevor Glasbey ,&nbsp;Zhuoran Chen ,&nbsp;Lia Moshkanbaryans ,&nbsp;Kate H. Moore ,&nbsp;Theerthankar Das ,&nbsp;Jim Manos","doi":"10.1016/j.bioflm.2025.100296","DOIUrl":"10.1016/j.bioflm.2025.100296","url":null,"abstract":"<div><div>Uropathogenic <em>Pseudomonas aeruginosa</em> is a significant contributor to catheter-associated urinary tract infections (CA-UTIs), distinguished by its unique biofilm-forming properties compared to other strains. Despite its clinical significance, optimized strategies for biofilm eradication in the bladder and on catheters remain limited. Thus, the aim of this study was to highlight the potent antibacterial and biofilm-inhibitory effects of N-acetyl cysteine (NAC) against uropathogenic <em>P. aeruginosa</em>. Additionally, we sought to investigate its effect against catheter obstruction caused by <em>P. aeruginosa</em> in a patient, and whether this phenomenon can be replicated <em>in vitro</em> to underscore the urgency of addressing this critical challenge.</div><div>We demonstrated that uropathogenic <em>P. aeruginosa</em> form thick, mucoid biofilms <em>in vitro</em> that can heavily occlude catheters, with bacterial titres of between 10⁸ and 10¹¹ CFU/cm, thus impairing catheter functionality. Furthermore, treatment with NAC significantly reduced viable bacteria by &gt; 4_log10 (p &lt; 0.01), and inhibited biofilm formation and associated obstruction till experiment endpoint (96h). NAC also displayed significant bactericidal activity (p &lt; 0.001) against <em>P. aeruginosa</em> and significantly impeded bacterial attachment and aggregation through modulation of colloidal forces and change in the structure of the bacterial cell surface, thus impairing the bacterium's ability to initiate biofilm development. Mechanistically, NAC alters the bacterial surface structure, disrupting biofilm-associated virulence.</div><div>Hence our study found that NAC treatment physically disrupts uropathogenic <em>P. aeruginosa</em> biofilms and significantly alters its virulence. Our novel findings highlight the dual bactericidal and anti-biofilm properties of NAC <em>in vitro</em>, offering valuable insights into its potential application for preventing <em>P. aeruginosa</em> biofilm formation and catheter blockage in CA-UTI management.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"10 ","pages":"Article 100296"},"PeriodicalIF":5.9,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144663355","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Asia-Pacific Biofilms 2024: A global conference on microbial biofilms","authors":"Zhenbo Xu ,&nbsp;Birthe Kjellerup ,&nbsp;Chuanwu Xi ,&nbsp;Enrico Marsili ,&nbsp;Gamini Seneviratne ,&nbsp;Guanglei Qiu ,&nbsp;Honghua Hu ,&nbsp;Luyan Ma ,&nbsp;Liang Yang ,&nbsp;Yue Qu ,&nbsp;Yulong Tan ,&nbsp;Yaqin Li ,&nbsp;Feifeng Zhong ,&nbsp;Junyan Liu","doi":"10.1016/j.bioflm.2025.100322","DOIUrl":"10.1016/j.bioflm.2025.100322","url":null,"abstract":"","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"10 ","pages":"Article 100322"},"PeriodicalIF":4.9,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145732867","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Potassium iodide enhances the antimicrobial activity of plasma-activated water","authors":"Laura A. McClenaghan ,&nbsp;Thomas P. Thompson ,&nbsp;Akash Shambharkar ,&nbsp;Ross M. Duncan ,&nbsp;Paula Bourke ,&nbsp;Timofey Skvortsov ,&nbsp;Brendan F. Gilmore","doi":"10.1016/j.bioflm.2025.100313","DOIUrl":"10.1016/j.bioflm.2025.100313","url":null,"abstract":"&lt;div&gt;&lt;div&gt;Plasma-activated water (PAW) is a promising disinfection strategy that generates a complex mixture of reactive oxygen and nitrogen species (ROS/RNS), including hydrogen peroxide (H&lt;sub&gt;2&lt;/sub&gt;O&lt;sub&gt;2&lt;/sub&gt;), nitrate (NO&lt;sub&gt;3&lt;/sub&gt;&lt;sup&gt;−&lt;/sup&gt;), and transient oxidants, in an acidic aqueous environment. These reactive species contribute to both immediate and extended antimicrobial activity. This study investigates how the addition of low concentrations (&lt;100 μM) of potassium iodide (KI) enhances the bactericidal properties of spark-generated PAW by enabling the in-situ generation of reactive iodine species (RIS), particularly hypoiodous acid (HIO), under acidic conditions.&lt;/div&gt;&lt;div&gt;KI addition (10–100 μM) led to a counterintuitive, dose-dependent increase in H&lt;sub&gt;2&lt;/sub&gt;O&lt;sub&gt;2&lt;/sub&gt; concentrations, from ∼1.2 mM in PAW alone to ∼1.8 mM at 30 μM KI, possibly due to iodine-mediated catalytic effects or reduced H&lt;sub&gt;2&lt;/sub&gt;O&lt;sub&gt;2&lt;/sub&gt; degradation. NO&lt;sub&gt;3&lt;/sub&gt;&lt;sup&gt;−&lt;/sup&gt; levels also increased by ∼17 % with increasing KI. Equivalent concentrations of H&lt;sub&gt;2&lt;/sub&gt;O&lt;sub&gt;2&lt;/sub&gt; + KI failed to replicate the rapid antimicrobial activity observed in PAW + KI, which achieved complete inactivation of &lt;em&gt;Escherichia coli&lt;/em&gt; and &lt;em&gt;Listeria monocytogenes&lt;/em&gt; planktonic cells within 3 min, compared to over 10 min for PAW alone, indicating the involvement of additional reactive species in KI-enhanced antimicrobial activity of PAW. However, &lt;em&gt;Salmonella enterica planktonic cells&lt;/em&gt; exhibited only partial inactivation even with KI, indicating species-specific tolerance under these conditions. 24h biofilms of &lt;em&gt;L. monocytogenes&lt;/em&gt; and &lt;em&gt;E. coli&lt;/em&gt; were eradicated with PAW + KI in 10 min, whereas &lt;em&gt;S. enterica&lt;/em&gt; showed only a 2-log reduction.&lt;/div&gt;&lt;div&gt;Scavenger assays revealed that both longer-lived species (H&lt;sub&gt;2&lt;/sub&gt;O&lt;sub&gt;2&lt;/sub&gt;) and shorter-lived oxidants such as singlet oxygen are essential for this enhanced killing, while ozone and superoxide appeared dispensable. These findings support a multi-step antimicrobial mechanism: (1) plasma treatment creates a low pH, H&lt;sub&gt;2&lt;/sub&gt;O&lt;sub&gt;2&lt;/sub&gt;-rich solution; (2) iodide is oxidised to RIS such as I&lt;sub&gt;3&lt;/sub&gt;&lt;sup&gt;−&lt;/sup&gt; and HIO; (3) additional PAW-derived oxidants potentiate RIS chemistry; and (4) unionised HIO diffuses across bacterial membranes to induce oxidative damage.&lt;/div&gt;&lt;div&gt;PAW-KI remained stable for at least 14 days at 4 °C, with sustained RIS activity and minimal loss of H&lt;sub&gt;2&lt;/sub&gt;O&lt;sub&gt;2&lt;/sub&gt; or NO&lt;sub&gt;3&lt;/sub&gt;&lt;sup&gt;−&lt;/sup&gt;, suggesting preserved antimicrobial capacity over time. The antimicrobial mechanism likely proceeds through a four-step pathway: plasma-mediated generation of H&lt;sub&gt;2&lt;/sub&gt;O&lt;sub&gt;2&lt;/sub&gt; and NO&lt;sub&gt;3&lt;/sub&gt;&lt;sup&gt;−&lt;/sup&gt;; oxidation of I&lt;sup&gt;−&lt;/sup&gt; to I&lt;sub&gt;2&lt;/sub&gt; and HIO; potentiation of RIS via PAW-derived ROS/RNS; and subsequent microbial inactivation via membrane damage.&lt;/div&gt;&lt;div&gt;Together, these results demonstrate that PAW + KI f","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"10 ","pages":"Article 100313"},"PeriodicalIF":4.9,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145018471","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Current knowledge on the polymicrobial interaction and biofilm between Saccharomyces and Lactobacillaceae: regulatory mechanisms and applications","authors":"Zhenbo Xu ,&nbsp;Mahesh Premarathna ,&nbsp;Yaqin Li ,&nbsp;Xiaomao Yin ,&nbsp;Thanapop Soteyome ,&nbsp;Junyan Liu ,&nbsp;Gamini Seneviratne","doi":"10.1016/j.bioflm.2025.100336","DOIUrl":"10.1016/j.bioflm.2025.100336","url":null,"abstract":"<div><div>The family <em>Lactobacillaceae</em> plays a crucial role in food fermentation and probiotic applications, and exhibiting metabolic versatility and adaptability to diverse nutrient-rich environments. They are abundant in nutrient-rich environments like fermented food, vegetables, and the vaginal and gastrointestinal tracts of animals, where they metabolize carbohydrates to produce lactic acids. They also produce bioactive compounds and exhibit anti-inflammatory, antibacterial, and antifungal properties. <em>Saccharomyces</em> yeasts are also widely applied in food, medicine, and biofuel industries. Some species, such as <em>S</em>. <em>boulardii</em>, are recognized for their probiotic benefits, particularly in promoting gut health and alleviating intestinal disorders. This review focuses on the polymicrobial interactions between <em>Lactobacillaceae</em> and <em>Saccharomyces</em> species, highlighting their synergistic roles in improving fermentation efficiency, product quality, and microbial stability through mechanisms such as biofilm formation, metabolic exchange, and nutrient sharing. We also discuss competitive interactions observed in certain systems, such as sugarcane fermentation, which demonstrate the complexity of microbial ecology and the need for precise microbial management strategies. By synthesizing current research, this review aims to provide a comprehensive understanding of how microbial interactions influence fermentation outcomes, and to identify existing knowledge gaps and future directions for optimizing industrial applications.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"10 ","pages":"Article 100336"},"PeriodicalIF":4.9,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145617920","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"EUROBIOFILMS 2024 - From environment and basic science to serious clinical infections - bridging for solutions","authors":"Claus Moser,&nbsp;Thomas Bjarnsholt,&nbsp;Trine Rolighed Thomsen","doi":"10.1016/j.bioflm.2025.100300","DOIUrl":"10.1016/j.bioflm.2025.100300","url":null,"abstract":"","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"10 ","pages":"Article 100300"},"PeriodicalIF":4.9,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145732868","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Corrigendum to “In vitro study to assess modulation of Candida biofilm by Escherichia coli from vaginal strains” [Biofilm 5 (2023) 100116]","authors":"Marta Díaz-Navarro ,&nbsp;Álvaro Irigoyen ,&nbsp;María Palomo ,&nbsp;Pilar Escribano ,&nbsp;Jesús Guinea ,&nbsp;Almudena Burillo ,&nbsp;Alicia Galar ,&nbsp;Patricia Muñoz ,&nbsp;María Guembe","doi":"10.1016/j.bioflm.2025.100297","DOIUrl":"10.1016/j.bioflm.2025.100297","url":null,"abstract":"","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"10 ","pages":"Article 100297"},"PeriodicalIF":4.9,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145736090","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Toward an improved in vitro model of prosthetic joint infection for Staphylococcus aureus biofilm characterization","authors":"Yasser Dghoughi ,&nbsp;Jennifer Varin-Simon ,&nbsp;Sophie C. Gangloff ,&nbsp;Marius Colin ,&nbsp;Fany Reffuveille","doi":"10.1016/j.bioflm.2025.100325","DOIUrl":"10.1016/j.bioflm.2025.100325","url":null,"abstract":"<div><div>Biofilm formation on orthopedic implants is often implicated in chronic prosthetic and joint infections (PJI) that are complex to manage. To date, no current bacterial <em>in vitro</em> model can fully simulate the PJI environment leading to a lack of knowledge to develop diagnosis tool and adapted treatment. Our project aims to set up an innovative <em>in vitro</em> model to characterize <em>Staphylococcus aureus</em> clinical strains biofilms in a PJI context, focusing on several parameters: culture media, incubation time, atmospheric conditions and support for biofilm growth.</div><div>Biofilm formation was evaluated in various culture media, by counting both planktonic and adherent bacteria (CFU) and quantifying biofilm biomass using crystal violet staining. A mature biofilm was obtained after 72 h of incubation with a similar proportion of planktonic and adherent bacteria whereas a variable dispersion was observed at 96 h.</div><div>Comparing two different oxygen concentrations (Hypoxia 2.5 % like in bone site <em>vs</em> Anoxia) revealed that a slight variation had a strong impact on biofilm formation, underlining the fact that the physiological conditions are highly necessary to set a mimetic model. A medium has therefore been developed, the modified Bone-Like Environment (BLE+) allowing a consistent biofilm growth.</div><div>When studying bacterial adhesion, planktonic bacteria can gather and form aggregates that are distinct from mature biofilms. To avoid this phenomenon, a suspended pegs was used. By holding the pegs in the medium, we specifically drove active bacterial adhesion related to biofilm formation, eliminating interference from sedimented aggregates. Moreover, to limit the interaction between planktonic bacteria and biofilm over the 72 h, a medium renewal was applied at 8 h of incubation with a low impact on biofilm biomass.</div><div>This method allowed the observation of differences between the USA300 (MRSA) and SH1000 (MSSA) strains: the MSSA showed more adherent bacteria and bigger aggregates than the MRSA strain.</div><div>In conclusion, the parameters for an <em>in vitro</em> biofilm model simulating PJI context have been validated. These parameters include 2.5 % dioxygen, BLE + supplementation, and 72-h incubation on suspended titanium pegs with a renewal media after a primo bacteria adhesion of 8 h.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"10 ","pages":"Article 100325"},"PeriodicalIF":4.9,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145319585","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterizing species-specific metabolic signatures in vaginal microbiota across planktonic and biofilm states","authors":"Smrutiti Jena ,&nbsp;Damilola Lawore ,&nbsp;Leopold N. Green ,&nbsp;Douglas K. Brubaker","doi":"10.1016/j.bioflm.2025.100330","DOIUrl":"10.1016/j.bioflm.2025.100330","url":null,"abstract":"<div><div>Bacterial vaginosis (BV) affects about 29 % of U.S. women, with higher rates in some groups and up to 50–69 % recurrence within a year. It increases the risk of STIs, pregnancy complications, and can cause significant discomfort and reduced quality of life. Prior studies mapped vaginal metabolomes, but individual microbial metabolite signatures remain poorly understood. Given that biofilms exhibit distinct metabolic requirements compared to planktonic cultures, analyzing biofilm vs. planktonic culture metabolites with their unique metabolic needs may uncover novel treatment targets. This study provides a comprehensive metabolomic comparison of key vaginal microbes—<em>Lactobacillus crispatus, Gardnerella vaginalis</em>, and <em>Lactobacillus iners</em> under both planktonic and biofilm conditions. Our analysis showed that metabolite production and consumption vary by microbe and growth mode. <em>G. vaginalis</em> formed biofilms in multiple media, with limited shared metabolic pathways between its biofilm types, indicating unique metabolic processes. Despite L. <em>crispatus</em> suspension and biofilm cultures sharing 142 consumed and 104 produced metabolites, the biofilm culture demonstrated a remarkable metabolic shift. Comparing all three species, we observed convergence in nutrient utilization, but divergence in metabolic outputs reflecting growth-specific adaptations, highlighting the importance of considering microbial state in vaginal microbiome studies.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"10 ","pages":"Article 100330"},"PeriodicalIF":4.9,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145617923","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}