{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,11]],"date-time":"2026-10-11T13:48:02Z","timestamp":1791726482152,"version":"4.3.4"},"reference-count":35,"publisher":"Public Library of Science (PLoS)","issue":"9","license":[{"start":{"date-parts":[[2025,9,10]],"date-time":"2025-09-10T00:00:00Z","timestamp":1757462400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>\n                    <jats:italic>Trichophyton rubrum<\/jats:italic>\n                    , a dermatophyte, demonstrates a notable ability to form mature biofilms on skin and associated surfaces, strengthening its resistance to antifungal agents. This characteristic poses intricate challenges in dermatological research and therapeutic strategies, underscoring the need for innovative approaches to effectively manage fungal infections. This work assessed the impact of the anti-biofilm enzymes, i.e., cellulase, protease, and amylase, individually and in combination, on the eradication and inhibition of\n                    <jats:italic>T. rubrum<\/jats:italic>\n                    biofilm. After 168 hours of incubation,\n                    <jats:italic>T. rubrum<\/jats:italic>\n                    biofilm formation matured, and the anti-biofilm enzymes significantly reduced the rate of biofilm development. The rates of biofilm inhibition and eradication for cellulase, protease, and amylase were 64%, 38%, and 28% at 72 hours of incubation, and 47%, 25.87%, and 17%, respectively, at 168 hours. However, the combined anti-biofilm enzymes (cellulase, protease, and amylase) had a 60.62% biofilm suppression rate. SEM analysis revealed marked reductions in\n                    <jats:italic>T. rubrum<\/jats:italic>\n                    conidial density, disrupted hyphal structures, and diminished biofilm adherence in enzyme-treated samples compared to untreated controls, visually supporting the inhibitory effect observed in quantitative assays. These morphological alterations indicate compromised fungal viability and structural disintegration of the biofilm matrix. Collectively, the SEM findings reinforce the therapeutic potential of enzyme-based strategies against dermatophytic biofilms. Additionally, these anti-biofilm enzymes have shown strong efficacy in reducing the exopolysaccharide (EPS) content and degrading the complex EPS matrix network. The\n                    <jats:italic>T. rubrum<\/jats:italic>\n                    biofilm treated with anti-biofilm enzymes, such as cellulase and protease, resulted in EPS degradation by FTIR analysis. The antibiofilm enzyme cellulase showed notable degradation of the beta 1\u20134 linkage within the glycosidic bond. A significant degradation is observed when\n                    <jats:italic>T. rubrum<\/jats:italic>\n                    biofilm is treated with combined enzymes (cellulase, protease, and amylase). The combined enzymatic treatment disrupted the EPS matrix, indicating its potential as an effective strategy for inhibiting biofilm formation by T. rubrum.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0331291","type":"journal-article","created":{"date-parts":[[2025,9,10]],"date-time":"2025-09-10T17:33:18Z","timestamp":1757525598000},"page":"e0331291","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":3,"title":["In vitro characterization of Trichophyton rubrum biofilm by combined anti-biofilm enzymes"],"prefix":"10.1371","volume":"20","author":[{"given":"Amna","family":"Rehman","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4527-0293","authenticated-orcid":true,"given":"Fatima","family":"Ismail","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ayesha","family":"Safdar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Muhammad","family":"Imran","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2025,9,10]]},"reference":[{"issue":"4","key":"pone.0331291.ref001","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1002\/jobm.201900675","article-title":"Recent advances in the diagnosis of dermatophytosis","volume":"60","author":"J Begum","year":"2020","journal-title":"J Basic Microbiol"},{"key":"pone.0331291.ref002","doi-asserted-by":"crossref","first-page":"1980","DOI":"10.3389\/fmicb.2020.01980","article-title":"A two-way road: antagonistic interaction between dual-species biofilms formed by Candida albicans\/Candida parapsilosis and Trichophyton rubrum","volume":"11","author":"LM Garcia","year":"2020","journal-title":"Front Microbiol"},{"issue":"1","key":"pone.0331291.ref003","first-page":"32","article-title":"Onychomycosis: an updated review","volume":"14","author":"AKC Leung","year":"2020","journal-title":"Recent Pat Inflamm Allergy Drug Discov"},{"key":"pone.0331291.ref004","doi-asserted-by":"crossref","first-page":"1","DOI":"10.7573\/dic.2023-5-1","article-title":"Tinea pedis: an updated review","volume":"12","author":"AK Leung","year":"2023","journal-title":"Drug Context"},{"issue":"1","key":"pone.0331291.ref005","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1080\/14787210.2023.2144232","article-title":"Strategies to improve the diagnosis and clinical treatment of dermatophyte infections","volume":"21","author":"M Durdu","year":"2023","journal-title":"Expert Rev Anti Infect Ther"},{"issue":"1","key":"pone.0331291.ref006","doi-asserted-by":"crossref","first-page":"120","DOI":"10.1111\/ijd.16337","article-title":"Quantitative analysis of in\u2009vitro biofilm formation by clinical isolates of dermatophyte and antibiofilm activity of common antifungal drugs","volume":"62","author":"S Yazdanpanah","year":"2023","journal-title":"Int J Dermatol"},{"issue":"8","key":"pone.0331291.ref007","doi-asserted-by":"crossref","first-page":"878","DOI":"10.3390\/jof8080878","article-title":"Alternative splicing in Trichophyton rubrum Occurs in Efflux pump transcripts in response to antifungal drugs","volume":"8","author":"MER Lopes","year":"2022","journal-title":"J Fungi (Basel)"},{"key":"pone.0331291.ref008","article-title":"The efficacy of phage\u2013enzyme combinations to reduce Pseudomonas aeruginosa biofilms in 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Ohst","year":"2019","journal-title":"J Clin Microbiol"},{"issue":"1","key":"pone.0331291.ref024","doi-asserted-by":"crossref","first-page":"54","DOI":"10.4103\/0366-6999.172573","article-title":"A Trichophyton Rubrum Infection Model Based on the Reconstructed Human Epidermis - Episkin\u00ae","volume":"129","author":"P-P Liang","year":"2016","journal-title":"Chin Med J (Engl)"},{"issue":"3","key":"pone.0331291.ref025","first-page":"643","article-title":"Synergistic effects of cellulase and proteinase K on the disruption of Staphylococcus aureus biofilms","volume":"129","author":"G Zhou","year":"2020","journal-title":"J Appl Microbiol"},{"key":"pone.0331291.ref026","first-page":"249","author":"A Borges","year":"2020","journal-title":"Chapter 11 - Biofilm Control with Enzymes"},{"issue":"1","key":"pone.0331291.ref027","doi-asserted-by":"crossref","DOI":"10.1128\/spectrum.02589-21","article-title":"Hydrolytic enzymes as potentiators of antimicrobials against an inter-kingdom biofilm model","volume":"10","author":"A Ruiz-Sorribas","year":"2022","journal-title":"Microbiol Spectr"},{"issue":"2","key":"pone.0331291.ref028","doi-asserted-by":"crossref","first-page":"15","DOI":"10.3390\/microorganisms5020015","article-title":"Approaches to dispersing medical biofilms","volume":"5","author":"D Fleming","year":"2017","journal-title":"Microorganisms"},{"issue":"7","key":"pone.0331291.ref029","doi-asserted-by":"crossref","first-page":"783","DOI":"10.1080\/08927014.2020.1811856","article-title":"In vitro and ex vivo biofilms of dermatophytes: a new panorama for the study of antifungal drugs","volume":"36","author":"DDSCM Castelo-Branco","year":"2020","journal-title":"Biofouling"},{"issue":"8","key":"pone.0331291.ref030","doi-asserted-by":"crossref","first-page":"798","DOI":"10.1093\/mmy\/myv044","article-title":"Discovery of a sexual stage in Trichophyton onychocola, a presumed geophilic dermatophyte isolated from toenails of patients with a history of T. rubrum onychomycosis","volume":"53","author":"V Hubka","year":"2015","journal-title":"Med Mycol"},{"issue":"9","key":"pone.0331291.ref031","first-page":"5404","article-title":"Biochemical alterations of Candida albicans during the phenotypic transition from yeast to hyphae captured by Fourier transform mid-infrared\u2013attenuated reflectance spectroscopy","volume":"90","author":"Q-Y Shi","year":"2018","journal-title":"Anal Chem"},{"issue":"7","key":"pone.0331291.ref032","doi-asserted-by":"crossref","first-page":"1551","DOI":"10.3390\/molecules25071551","article-title":"Influence of the Sample Preparation Method in Discriminating Candida spp. Using ATR-FTIR Spectroscopy","volume":"25","author":"S Pebotuwa","year":"2020","journal-title":"Molecules"},{"key":"pone.0331291.ref033","first-page":"100141","article-title":"Fourier transform infrared spectroscopy as a label\u2011free, non\u2011destructive approach to monitoring biofilm progression","volume":"6","author":"B Short","year":"2023","journal-title":"Biofilm"},{"issue":"3","key":"pone.0331291.ref034","first-page":"222","article-title":"Biofilm formation in dermatophytes: Mechanism, clinical relevance, and treatment strategies","volume":"63","author":"M Sharma","year":"2020","journal-title":"Mycoses"},{"issue":"11","key":"pone.0331291.ref035","first-page":"754","article-title":"In vitro characterization of Trichophyton rubrum biofilm","volume":"57","author":"CB Costa-Orlandi","year":"2014","journal-title":"Mycoses"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0331291","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,9,10]],"date-time":"2025-09-10T17:33:26Z","timestamp":1757525606000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0331291"}},"subtitle":[],"editor":[{"given":"Noor","family":"Jahan","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2025,9,10]]},"references-count":35,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2025,9,10]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0331291","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,9,10]]}}}