Biofilm最新文献

{"title":"Squalamine and claramine A1 disperse Pseudomonas aeruginosa biofilm","authors":"Anne-Sophie Tareau ,&nbsp;Ali Tahrioui ,&nbsp;Mathieu Gonzalez ,&nbsp;Evan Croize ,&nbsp;Jennifer Varin Simon ,&nbsp;Magalie Barreau ,&nbsp;Audrey David ,&nbsp;Fany Reffuveille ,&nbsp;Jean-Michel Brunel ,&nbsp;Olivier Lesouhaitier ,&nbsp;Sylvie Chevalier","doi":"10.1016/j.bioflm.2025.100293","DOIUrl":"10.1016/j.bioflm.2025.100293","url":null,"abstract":"<div><div><em>Pseudomonas aeruginosa</em> is an opportunistic pathogen that causes both acute and chronic infections, including pneumonia, bloodstream infections, urinary tract infections, and surgical site infections. It poses a significant threat to individuals with chronic lung conditions, particularly those with cystic fibrosis. Squalamine and claramine A1 have emerged as promising antibacterial compounds, exhibiting activity against a broad range of both Gram-positive and Gram-negative bacteria. Beyond their potent antibacterial properties, our findings reveal that sub-inhibitory concentrations of claramine A1 and squalamine can disperse pre-formed <em>P. aeruginosa</em> biofilm without impacting bacterial growth. While claramine A1, but not squalamine, enhances membrane fluidity, the structural difference between these compounds lies primarily in their spermine and spermidine moieties. Notably, we found that spermine, unlike spermidine, was able to both disperse biofilm and increase membrane fluidity. Together, our results suggest that while both compounds are effective at disrupting <em>P. aeruginosa</em> biofilm, they likely act through distinct mechanisms.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"9 ","pages":"Article 100293"},"PeriodicalIF":5.9,"publicationDate":"2025-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144166736","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":"Targeting Staphylococcus aureus biofilm-related infections on implanted material with a novel dual-action thermosensitive hydrogel containing vancomycin and a tri-enzymatic cocktail: in vitro and in vivo studies","authors":"Randy Buzisa Mbuku ,&nbsp;Hervé Poilvache ,&nbsp;Loïc Maigret ,&nbsp;Rita Vanbever ,&nbsp;Françoise Van Bambeke ,&nbsp;Olivier Cornu","doi":"10.1016/j.bioflm.2025.100288","DOIUrl":"10.1016/j.bioflm.2025.100288","url":null,"abstract":"<div><div>Implant-associated infections remain a critical challenge due to the presence of biofilm-forming bacteria, which enhance tolerance to conventional treatments. This study investigates the efficacy of a tri-enzymatic cocktail (TEC; DNA/RNA endonuclease, endo-14-β-<span>d</span>-glucanase, β-<em>N</em>-acetylhexosaminidase) targeting biofilm matrix components combined with supratherapeutic doses of antibiotics encapsulated in a thermosensitive hydrogel (poloxamer P407) for local administration. <em>In vitro,</em> the hydrogel formulation enabled controlled release of active agents over 12 h. Vancomycin and TEC co-formulated in hydrogel achieved up to 3.8 Log₁₀ CFU count reduction and 80 % biofilm biomass reduction on MRSA biofilms grown on titanium coupons, demonstrating enhanced efficacy as compared to individual active agents, with 1.3–3.2 log₁₀ additional killing. Fluoroquinolone efficacy remained unchanged by enzyme addition. <em>In vivo,</em> in a model of tissue cages containing titanium beads implanted in the back of guinea pigs, hydrogel-delivered vancomycin maintained therapeutic levels for seven days. Coupled with an intraperitoneal administration of vancomycin for 4 days, a single local administration of hydrogel containing both vancomycin and TEC was more effective than hydrogels containing either vancomycin or TEC, achieving an additional 2.1 Log₁₀ CFU reduction compared to local vancomycin, 2.3 Log₁₀ compared to local TEC, and 4.3 Log₁₀ compared to systemic vancomycin treatment alone. However, partial regrowth occurred at later stages, indicating room for further optimization. Nevertheless, these findings already underscore the potential of combining a high dose of antibiotic with an enzymatic cocktail in a sustained-release hydrogel delivery system as a promising strategy for improving the management of biofilm-associated implant infections.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"9 ","pages":"Article 100288"},"PeriodicalIF":5.9,"publicationDate":"2025-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144124259","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":"Assessing the impact of UV-C exposure on pre-existing cultured marine diatom biofilms","authors":"Kailey N. Richard ,&nbsp;Andrew Palmer ,&nbsp;Geoffrey Swain ,&nbsp;Kelli Z. Hunsucker","doi":"10.1016/j.bioflm.2025.100285","DOIUrl":"10.1016/j.bioflm.2025.100285","url":null,"abstract":"<div><div>Ultraviolet-C (UV–C) has been considered as a potential tool in the maintenance and prevention of marine biofouling. However, it is unclear how irradiation disturbs biofouling, specifically biofilms, or the duration and frequency needed to reduce or eliminate their growth. The marine diatom, <em>Navicula incerta</em>, was used in a two-part lab study to determine the minimum time between UV-C duty cycling (also known as interval testing) that would result in the reduction and prevention of marine biofilms. The first study assessed the diatom's metabolic activity, morphology, behavior, and recovery to understand the influence of UV-C on the biofilm. Cell viability, biofilm matrixes, and chlorophyll <em>a</em> were significantly reduced immediately following UV-C exposure, yet when exposure was ceased biofilms began to recover. Based on these results, the second study investigated the duty cycle duration required to limit the growth of the cultured biofilm. Duty cycle testing was able to initially deplete cell viability and chlorophyll <em>a</em>; however, in between exposures, recovery was evident. These findings indicate that shorter (10 min; 1.86 J/cm²) doses of UV-C exposures must be applied more frequently than more prolonged doses (60 min; 11.16 J/cm²) to keep biofilm formation minimal. The duty cycles established here also provide a guideline of efficacy for future studies that can be applied to biofilms in both laboratory and the field.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"9 ","pages":"Article 100285"},"PeriodicalIF":5.9,"publicationDate":"2025-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144124341","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":"Protecting bacteriophages under UV irradiation with brilliant blue FCF for targeted bacterial control","authors":"Mateusz Wdowiak ,&nbsp;Aneta Magiera ,&nbsp;Magdalena Tomczyńska ,&nbsp;Witold Adamkiewicz ,&nbsp;Francesco Stellacci ,&nbsp;Jan Paczesny","doi":"10.1016/j.bioflm.2025.100286","DOIUrl":"10.1016/j.bioflm.2025.100286","url":null,"abstract":"<div><div>Compared to the standard methods for treating bacterial diseases, bacteriophages are eco-friendly and chemical-free. Exposure to ultraviolet (UV) light or sunlight hampers the efficacy of phage-based approaches. This is crucial when phages are <em>i)</em> exposed to sunlight (e.g., in agriculture) or <em>ii</em>) are to be used simultaneously with UV for sterilization. Here, we develop a method utilizing a food dye, brilliant blue FCF (BB), that selectively stabilizes bacteriophages against exposure to UV irradiation. In the absence of BB, all tested phages and bacteria are completely inactivated by UV exposure. However, with the addition of BB, all tested non-enveloped phages are effectively protected, while gram-negative bacteria remain vulnerable to UV inactivation. The mechanism of protection requires selective binding of BB to the virion. The simultaneous action of BB-stabilized bacteriophages and UV allows for the removal of up to 99.99 % of bacteria within only 30–60 min. We demonstrate the method's applicability in combating biofouling of membranes and food sterilization. We envision using the developed approach against biofouling in industrial processes, agriculture, and the food industry.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"9 ","pages":"Article 100286"},"PeriodicalIF":5.9,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143948733","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":"Harnessing emergent properties of microbial consortia for Agriculture: Assembly of the Xilonen SynCom","authors":"Gabriela Gastélum ,&nbsp;Bruno Gómez-Gil ,&nbsp;Gabriela Olmedo-Álvarez ,&nbsp;Jorge Rocha","doi":"10.1016/j.bioflm.2025.100284","DOIUrl":"10.1016/j.bioflm.2025.100284","url":null,"abstract":"<div><div>Synthetic communities (SynComs) are valuable tools for addressing microbial community assembly and function, towards their manipulation for clinical, biotechnological and agricultural applications. However, SynCom design is complicated since interactions between microbes cannot be predicted based on their individual properties. Here we aimed to assemble a functionally cohesive SynCom displaying high-order interactions, as a model to study the community-level beneficial functions of seed-endophytic bacteria from native maize landraces<em>,</em> including strains from the Bacilli class, and the <em>Burkholderia</em> and <em>Pseudomonas</em> genera. We developed a partial combinatorial, bottom-up strategy that was followed by the detection of complex colony architecture as an emergent property in co-cultures. Using this simplified approach, we tested less than 400 co-cultures from a pool of 27 strains, resulting in the assembly the <em>Xilonen</em> SynCom, which includes <em>Bacillus pumilus</em> NME155, <em>Burkholderia contaminans</em> XM7 and <em>Pseudomonas</em> sp. GW6. In this community, higher-order interactions result in complex colony architecture, which is considered a proxy of biofilm formation. Additionally, we generated protocols for absolute quantification of each member from a complex mixture. The <em>Xilonen</em> SynCom will serve as a model to study biofilm formation in community settings, and will aid in the study of the molecular and ecological basis mediating maize fertility.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"9 ","pages":"Article 100284"},"PeriodicalIF":5.9,"publicationDate":"2025-05-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143907643","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":"Machine learning assisted classification of staphylococcal biofilm maturity","authors":"S.C.J. van Dun ,&nbsp;R. Knol ,&nbsp;A.S. Silva-Herdade ,&nbsp;A.S. Veiga ,&nbsp;M.A.R.B. Castanho ,&nbsp;P.H. Nibbering ,&nbsp;B.G.C.W. Pijls ,&nbsp;A.M. van der Does ,&nbsp;J. Dijkstra ,&nbsp;M.G.J. de Boer","doi":"10.1016/j.bioflm.2025.100283","DOIUrl":"10.1016/j.bioflm.2025.100283","url":null,"abstract":"<div><div>An increasing incidence of device-related, biofilm-associated infections has been observed in clinical practice worldwide. <em>In vitro</em> biofilm models are essential to study these burdensome infections and to design and test potential new treatment approaches. However, there is considerable variation in <em>in vitro</em> biofilm models, and a generally accepted systematic description of biofilm maturity – apart from incubation time – is lacking.</div><div>Therefore, we proposed a scheme comprised of 6 different classes based on common topographic characteristics, <em>i.e.</em>, the substrate, bacterial cells and extracellular matrix, identified by atomic force microscopy (AFM), to describe biofilm maturity independent of incubation time. Evaluation of a test set of staphylococcal biofilm images by a group of independent researchers showed that human observers were capable of classifying images with a mean accuracy of 0.77 ± 0.18. However, manual evaluation of AFM biofilm images is time-consuming, and subject to observer bias. To circumvent these disadvantages, a machine learning algorithm was designed and developed to aid in classification of biofilm images.</div><div>The designed algorithm was capable of identifying pre-set characteristics of biofilms and able to discriminate between the six different classes in the proposed framework. Compared to the established ground truth, the mean accuracy of the developed algorithm amounted to 0.66 ± 0.06 with comparable recall, and off-by-one accuracy of 0.91 ± 0.05. This algorithm, which classifies AFM images of biofilms, has been made available as an open access desktop tool.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"9 ","pages":"Article 100283"},"PeriodicalIF":5.9,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143907642","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":"PMA-qPCR to quantify viable cells in multispecies oral biofilm after disinfectant treatments","authors":"Sybille Schwendener,&nbsp;Manuela Flury,&nbsp;Joël Jenzer,&nbsp;Thomas Thurnheer,&nbsp;Lamprini Karygianni","doi":"10.1016/j.bioflm.2025.100281","DOIUrl":"10.1016/j.bioflm.2025.100281","url":null,"abstract":"<div><div>Conventional quantitative real-time PCR (qPCR) amplifies DNA from viable and dead cells, which can lead to an overestimation of live bacteria. Viability qPCR aims to eliminate DNA from membrane-compromised cells through treatment with propidium monoazide (PMA).</div><div>Here, we evaluated PMA-qPCR to enumerate viable cells of <em>Actinomyces oris</em>, <em>Fusobacterium nucleatum</em>, <em>Streptococcus oralis</em>, <em>Streptococcus mutans,</em> and <em>Veillonella dispar</em>. Five-species oral biofilms were grown on hydroxyapatite discs for 64 h. The biofilms were exposed to 0.2 % chlorhexidine (CHX) or 3 % sodium hypochlorite (NaOCl) for 2 min, either once before cell harvest at 64 h or six times during biofilm growth. The total and single species cells were quantified by culture (CFU) and qPCR from samples with and without PMA treatment before DNA extraction. For species-specific qPCR, TaqMan assays were applied. To determine total bacteria counts, a SYBR green qPCR was established using universal degenerative primers for the conserved <em>dnaK</em> gene.</div><div>For biofilms treated once with CHX, the addition of PMA led to a 1 to 1.6 log₁₀ reduction in PCR counts. This closely matched CFU and PMA-qPCR counts for total bacteria and all single species, except for <em>F. nucleatum</em>, where PMA-qPCR detected significantly more bacteria than culture. NaOCl treatment directly affected DNA and inhibited subsequent PCR amplification, even in samples without PMA. Single treatment of biofilms with 3 % NaOCl and six-fold exposure of biofilms to disinfectants resulted in no viable cell detection by culture. However, PMA did not completely prevent PCR amplification, indicating that disinfectant efficacy measured by viability PCR could be underestimated.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"9 ","pages":"Article 100281"},"PeriodicalIF":5.9,"publicationDate":"2025-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143859281","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":"Efficacy of nitric oxide donors and EDTA against Pseudomonas aeruginosa biofilms: Implications for antimicrobial therapy in chronic wounds","authors":"Aaron Crowther ,&nbsp;Gareth LuTheryn ,&nbsp;Ramón Garcia-Maset ,&nbsp;Maryam Parhizkar ,&nbsp;J. Mark Sutton ,&nbsp;Charlotte Hind ,&nbsp;Dario Carugo","doi":"10.1016/j.bioflm.2025.100280","DOIUrl":"10.1016/j.bioflm.2025.100280","url":null,"abstract":"<div><div>Opportunistic pathogen <em>Pseudomonas aeruginosa</em> plays a crucial role in chronic wound biofilms, increasing infection's morbidity and mortality. In recent years, the signalling molecule nitric oxide (NO) and chelating agent tetrasodium EDTA (T-EDTA) have been applied therapeutically owing to their multifactorial effects including bacterial killing, biofilm dispersal, and wound healing. However, previous studies assessing NO's antibiofilm efficacy have not considered the variable pH and temperature of the wound environment. Here, pH-dependent NO donors <em>N</em>-diazeniumdiolates (NONOates), PAPA NONOate (PA-NO) and Spermine NONOate (SP–NO), and T-EDTA were applied in wound-relevant pH environments (pH 5.5–8.5) and temperatures (32 °C and 37 °C) to <em>P. aeruginosa</em> PAO1 biofilms grown for either 24 or 48 h. At 32 °C and pH 7.5, 250 μM PA-NO reduced 24-h biofilm biomass by 35 %. At 37 °C, 250 μM PA-NO and 4 % w/v T-EDTA caused 21 % and 57 % biomass reduction in 24-h biofilms, respectively. In 48-h biofilms, NONOates did not induce significant biomass reduction, while T-EDTA maintained its efficacy with a 64 % reduction. A subsequent experiment investigated the impact of NONOates and T-EDTA as pre-treatments before exposure to ciprofloxacin. Unexpectedly, NONOate pre-treatment decreased ciprofloxacin's effectiveness, resulting in approximately 1-log increase in viable planktonic and biofilm-residing cells compared to ciprofloxacin alone. It was hypothesized that this protective effect might stem from NO-induced decreased cellular respiration, which inhibits reactive oxygen species (ROS)-mediated bactericidal mechanisms. These findings highlight both the potential and complexities of developing effective antimicrobial strategies for chronic wound infections, emphasizing the need for further research to optimize treatment approaches.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"9 ","pages":"Article 100280"},"PeriodicalIF":5.9,"publicationDate":"2025-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143877455","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":"Modeling reciprocal adaptation of Staphylococcus aureus and Pseudomonas aeruginosa co-isolates in artificial sputum medium","authors":"Zhifen Wang ,&nbsp;Emily Giedraitis ,&nbsp;Christiane Knoop ,&nbsp;Daniel J. Breiner ,&nbsp;Vanessa V. Phelan ,&nbsp;Françoise Van Bambeke","doi":"10.1016/j.bioflm.2025.100279","DOIUrl":"10.1016/j.bioflm.2025.100279","url":null,"abstract":"<div><div>Co-infections by <em>Staphylococcus aureus</em> and <em>Pseudomonas aeruginosa</em> are frequent in the airways of patients with cystic fibrosis. These co-infections show higher antibiotic tolerance <em>in vitro</em> compared to mono-infections. <em>In vitro</em> models have been developed to study the interspecies interactions between <em>P. aeruginosa</em> and <em>S. aureus</em>. However, these model systems fail to incorporate clinical isolates with diverse phenotypes, do not reflect the nutritional environment of the CF airway mucus, and/or do not model the biofilm mode of growth observed in the CF airways. Here, we established a dual-species biofilm model grown in artificial sputum medium, where <em>S. aureus</em> was inoculated before <em>P. aeruginosa</em> to facilitate the maintenance of both species over time. It was successfully applied to ten pairs of clinical isolates exhibiting different phenotypes. Co-isolates from individual patients led to robust, stable co-cultures, supporting the theory of cross-adaptation <em>in vivo</em>. Investigation into the cross-adaptation of the VBB496 co-isolate pair revealed that both the <em>P. aeruginosa</em> and <em>S. aureus</em> isolates had reduced antagonism, in part due to reduced production of <em>P. aeruginosa</em> secondary metabolites as well as higher tolerance to those metabolites by <em>S. aureus</em>. Together, these results indicate that the two-species biofilm model system provides a useful tool for exploring interspecies interactions of <em>P. aeruginosa</em> and <em>S. aureus</em> in the context of CF airway infections.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"9 ","pages":"Article 100279"},"PeriodicalIF":5.9,"publicationDate":"2025-04-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143850345","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 burden of antimicrobial resistance in biofilm forming Staphylococcus spp. from Vernal Keratoconjunctivitis patients eyes","authors":"Nelaveni Rupa ,&nbsp;Pragnya Rao Donthineni ,&nbsp;Sayan Basu ,&nbsp;Kotakonda Arunasri","doi":"10.1016/j.bioflm.2025.100278","DOIUrl":"10.1016/j.bioflm.2025.100278","url":null,"abstract":"<div><div>Vernal keratoconjunctivitis (VKC) is a chronic allergic ocular surface disease with seasonal recurrences and severe forms showing vision threatening complications. The purpose of the study is to understand the prevalence and diversity of biofilm-forming bacteria and antimicrobial resistance in VKC compared to healthy individuals (HC). For this, conjunctival swab samples were collected from VKC (n = 26) and HC (n = 23), of which culture positive samples were 77 % and 78.26 % respectively. The 16S rRNA gene sequencing revealed a significant increase in bacterial diversity in VKC compared to HC (p &lt; 0.05), identifying 16 and 9 bacterial species, respectively. <em>Staphylococcus epidermidis</em> emerged as the predominant bacterium in both groups, with relative abundances of 52.8 % in HC and 30.2 % in VKC (p &lt; 0.001). Biofilm formation was observed in 64.15 % of bacterial species in VKC and 31 % in HC (p &lt; 0.001). Scanning electron microscopy analysis confirmed temporal biofilm formation by <em>Staphylococcus</em> spp. in both groups. Minimum inhibitory concentration testing showed that biofilm forming <em>Staphylococcus</em> spp. from VKC exhibited multidrug resistance (&gt;2 antibiotics) more frequently than those from HC. Additionally, <em>Staphylococcus</em> spp. in VKC demonstrated higher resistance to fluoroquinolones compared to HC. These findings indicate a significantly greater prevalence of biofilm-forming and antimicrobial resistant <em>Staphylococcus</em> bacteria in VKC Patients compared with HC.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"9 ","pages":"Article 100278"},"PeriodicalIF":5.9,"publicationDate":"2025-04-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143820705","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}