{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,28]],"date-time":"2026-04-28T01:31:54Z","timestamp":1777339914548,"version":"3.51.4"},"reference-count":36,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2022,2,3]],"date-time":"2022-02-03T00:00:00Z","timestamp":1643846400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Conselho Nacional de Desenvolvimento Cient\u0131\u0301fico e Tecnolo\u0301gico","award":["xxx"],"award-info":[{"award-number":["xxx"]}]},{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfeicoamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","award":["xxx"],"award-info":[{"award-number":["xxx"]}],"id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Funda\u00e7\u00e3o Carlos Chagas Filho de Amparo \u00e0 Pesquisa do Estado do Rio de Janeiro Carlos Chagas Filho","award":["xxx"],"award-info":[{"award-number":["xxx"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Sporotrichosis is the most prevalent subcutaneous mycosis globally, and it is typically caused by direct inoculation of the soil saprophytic fungus Sporothrix spp. into the patients\u2019 skin. However, sporotrichosis has an important zoonotic transmission route between cats and humans in hot-spot endemic areas such as Brazil. Antifungal itraconazole is the first-line treatment; however, it is frequently associated with recurrence after withdrawal, mainly on cats. Biofilms are important resistance structures related to the environmental persistence of most microorganisms. In the present work, we evaluated Sporothrix yeasts\u2019 ability to form biofilms in an ex vivo model of infected claws of cats. Using scanning electron microscopy, we demonstrated the presence of fungal biofilms in the claws of cats diagnosed with sporotrichosis confirmed by isolation of Sporothrix spp. in culture. We present here evidence of antibiofilm activity of miltefosine and suggest its use off-label as an antifungal as a putative alternative to itraconazole against Sporothrix biofilms. Claw contamination could sustain infections through a continuous inoculation cycle between open lesions and cat claws. Our results further support the off-label use of miltefosine as a promising alternative, especially for mycosis refractory to conventional treatment.<\/jats:p>","DOI":"10.3390\/pathogens11020206","type":"journal-article","created":{"date-parts":[[2022,2,6]],"date-time":"2022-02-06T20:38:40Z","timestamp":1644179920000},"page":"206","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":24,"title":["Sporothrix spp. Biofilms Impact in the Zoonotic Transmission Route: Feline Claws Associated Biofilms, Itraconazole Tolerance, and Potential Repurposing for Miltefosine"],"prefix":"10.3390","volume":"11","author":[{"given":"Giulia Maria Pires","family":"dos Santos","sequence":"first","affiliation":[{"name":"Programa de Biologia Celular e Parasitologia, Instituto de Biof\u00edsica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, RJ, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6576-6646","authenticated-orcid":false,"given":"Luana Pereira","family":"Borba-Santos","sequence":"additional","affiliation":[{"name":"Programa de Biologia Celular e Parasitologia, Instituto de Biof\u00edsica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, RJ, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Taissa","family":"Vila","sequence":"additional","affiliation":[{"name":"Programa de Biologia Celular e Parasitologia, Instituto de Biof\u00edsica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, RJ, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8428-7616","authenticated-orcid":false,"given":"Isabella Dib","family":"Ferreira Gremi\u00e3o","sequence":"additional","affiliation":[{"name":"Instituto Nacional de Infectologia Evandro Chagas, Funda\u00e7\u00e3o Oswaldo Cruz, Rio de Janeiro 21040-360, RJ, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0614-4713","authenticated-orcid":false,"given":"Sandro Antonio","family":"Pereira","sequence":"additional","affiliation":[{"name":"Instituto Nacional de Infectologia Evandro Chagas, Funda\u00e7\u00e3o Oswaldo Cruz, Rio de Janeiro 21040-360, RJ, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wanderley","family":"De Souza","sequence":"additional","affiliation":[{"name":"Programa de Biologia Celular e Parasitologia, Instituto de Biof\u00edsica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, RJ, Brazil"},{"name":"Centro Nacional de Biologia Estrutural e Bioimagem (CENABIO), Cidade Universit\u00e1ria, Rio de Janeiro 21941-902, RJ, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3659-5945","authenticated-orcid":false,"given":"Sonia","family":"Rozental","sequence":"additional","affiliation":[{"name":"Programa de Biologia Celular e Parasitologia, Instituto de Biof\u00edsica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, RJ, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,2,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Rossow, J.A., Queiroz-Telles, F., Caceres, D.H., Beer, K.D., Jackson, B.R., Pereira, J.G., Ferreira Gremi\u00e3o, I.D., and Pereira, S.A. (2020). A One Health Approach to Combatting Sporothrix brasiliensis: Narrative Review of an Emerging Zoonotic Fungal Pathogen in South America. J. Fungi, 6.","DOI":"10.3390\/jof6040247"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"606","DOI":"10.1590\/abd1806-4841.2017279","article-title":"Sporotrichosis: An update on epidemiology, etiopathogenesis, laboratory and clinical therapeutics","volume":"92","author":"Macedo","year":"2017","journal-title":"An. Bras. Dermatol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"813","DOI":"10.1007\/s11046-020-00425-0","article-title":"The threat of emerging and re-emerging pathogenic Sporothrix species","volume":"185","author":"Rodrigues","year":"2020","journal-title":"Mycopathologia"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"621","DOI":"10.3201\/eid2603.190803","article-title":"Geographic Expansion of Sporotrichosis, Brazil","volume":"26","author":"Oliveira","year":"2020","journal-title":"Emerg. Infect. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1093\/mmy\/myx043","article-title":"Antifungal susceptibility of Sporothrix schenckii complex biofilms","volume":"56","author":"Brilhante","year":"2018","journal-title":"Med. Mycol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1080\/mmy.39.1.147.149","article-title":"Isolation of Sporothrix schenckii from the nails of domestic cats (Felis catus)","volume":"39","author":"Schubach","year":"2001","journal-title":"Med. Mycol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"372","DOI":"10.1590\/S1517-83822006000300031","article-title":"Isolation of Sporothrix schenkii from the nails of healthy cats","volume":"37","author":"Souza","year":"2006","journal-title":"Braz. J. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1007\/s11046-013-9658-8","article-title":"Isolation of Sporothrix schenckii from the claws of domestic cats (indoor and outdoor) and in captivity in S\u00e3o Paulo (Brazil)","volume":"176","author":"Borges","year":"2013","journal-title":"Mycopathologia"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"107947","DOI":"10.1016\/j.exppara.2020.107947","article-title":"Comparative study of the use of miltefosine, miltefosine plus allopurinol, and allopurinol in dogs with visceral leishmaniasis","volume":"217","author":"Dias","year":"2020","journal-title":"Exp. Parasitol."},{"key":"ref_10","unstructured":"(2021, December 21). Leishmaniose Tegumentar (LT), Available online: https:\/\/www.gov.br\/saude\/pt-br\/assuntos\/saude-de-a-a-z\/l\/leishmaniose-tegumentar-lt."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"320","DOI":"10.1093\/mmy\/myt027","article-title":"In vitro inhibitory effect of miltefosine against strains of Histoplasma capsulatum var. capsulatum and Sporothrix spp.","volume":"52","author":"Brilhante","year":"2014","journal-title":"Med. Mycol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"962","DOI":"10.1111\/jam.12891","article-title":"In vitro antifungal activity of miltefosine and levamisole: Their impact on ergosterol biosynthesis and cell permeability of dimorphic fungi","volume":"119","author":"Brilhante","year":"2015","journal-title":"J. Appl. Microbiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1099\/jmm.0.000041","article-title":"Miltefosine is active against Sporothrix brasiliensis isolates with in vitro low susceptibility to amphotericin B or itraconazole","volume":"64","author":"Gagini","year":"2015","journal-title":"J. Med. Microbiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1016\/j.ijantimicag.2016.12.020","article-title":"Miltefosine is fungicidal to Paracoccidioides spp. yeast cells but subinhibitory concentrations induce melanisation","volume":"49","author":"Rossi","year":"2017","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1086\/381200","article-title":"Cat-transmitted sporotrichosis epidemic in Rio de Janeiro, Brazil: Description of a series of cases","volume":"38","author":"Barros","year":"2004","journal-title":"Clin. Infect. Dis."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"104670","DOI":"10.1016\/j.micpath.2020.104670","article-title":"Biofilm formation on cat claws by Sporothrix species: An ex vivo model","volume":"150","author":"Brilhante","year":"2021","journal-title":"Microb. Pathog."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"De Miranda, L.H.M., Silva, J.N., Gremi\u00e3o, I.D.F., Menezes, R.C., Almeida-Paes, R., Dos Reis, \u00c9.G., De Oliveira, R.D.V.C., De Araujo, D.S.d.A., Ferreiro, L., and Pereira, S.A. (2018). Monitoring Fungal Burden and Viability of Sporothrix spp. in Skin Lesions of Cats for Predicting Antifungal Treatment Response. J. Fungi, 4.","DOI":"10.3390\/jof4030092"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1093\/mmy\/myz106","article-title":"Molecular epidemiology of Sporothrix schenkii isolates in Malaysia","volume":"58","author":"Selvarajah","year":"2020","journal-title":"Med. Mycol."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Borba-Santos, L.P., Vila, T., and Rozental, S. (2020). Identification of two potential inhibitors of Sporothrix brasiliensis and Sporothrix schenckii in the Pathogen Box collection. PLoS ONE, 15.","DOI":"10.1371\/journal.pone.0240658"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"437","DOI":"10.1002\/jobm.3620290712","article-title":"Keratinophilic fungi associated with rabbit claws in Egypt","volume":"29","author":"Moharram","year":"1989","journal-title":"J. Basic Microbiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1007\/BF02837919","article-title":"Keratinophilic Fungi: Nature\u2019s keratin degrading machines! their isolation, identification and ecological role","volume":"8","author":"Sharma","year":"2003","journal-title":"Resonance"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1465","DOI":"10.2217\/fmb-2019-0302","article-title":"Comparative genomics of Sporothrix species and identification of putative pathogenic-gene determinants","volume":"12","author":"Prakash","year":"2020","journal-title":"Future Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"400","DOI":"10.1128\/MMBR.67.3.400-428.2003","article-title":"Candida albicans secreted aspartyl proteinases in virulence and pathogenesis","volume":"67","author":"Naglik","year":"2003","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"00312","DOI":"10.1128\/AAC.00312-18","article-title":"Miltefosine Has a Postantifungal Effect and Induces Apoptosis in Cryptococcus Yeasts","volume":"62","author":"Spadari","year":"2018","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"854","DOI":"10.3389\/fmicb.2020.00854","article-title":"Antifungal Activity and Mode of Action of Miltefosine Against Clinical Isolates of Candida krusei","volume":"11","author":"Wu","year":"2020","journal-title":"Front. Microbiol."},{"key":"ref_26","unstructured":"Sultan, S. (2016). Biofilm Formation as a Pathogenic Factor of Medically Important Fungi. Fungal Pathogenicity, IntechOpen. Chapter 1."},{"key":"ref_27","first-page":"456","article-title":"New Targets for the Development of Antifungal Agents","volume":"Volume 1","author":"Zaragoza","year":"2021","journal-title":"Encyclopedia of Mycology"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"698662","DOI":"10.3389\/fcimb.2021.698662","article-title":"Miltefosine Against Scedosporium and Lomentospora Species: Antifungal Activity and Its Effects on Fungal Cells","volume":"11","author":"Almeida","year":"2021","journal-title":"Front. Cell Infect. Microbiol."},{"key":"ref_29","first-page":"1571","article-title":"Miltefosine Administration in Cats with Refractory Sporotrichosis","volume":"46","author":"Silva","year":"2018","journal-title":"Acta Sci. Vet."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Rodrigues, A.M., de Melo Teixeira, M., de Hoog, G.S., Schubach, T.M., Pereira, S.A., Fernandes, G.F., Bezerra, L.M., Felipe, M.S., and de Camargo, Z.P. (2013). Phylogenetic analysis reveals a high prevalence of Sporothrix brasiliensis in feline sporotrichosis outbreaks. PLoS Negl. Trop. Dis., 7.","DOI":"10.1371\/journal.pntd.0002281"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"843","DOI":"10.3389\/fmicb.2017.00843","article-title":"Sporothrix schenckii sensu stricto and Sporothrix brasiliensis Are Differentially Recognized by Human Peripheral Blood Mononuclear Cells","volume":"8","year":"2017","journal-title":"Front. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"681","DOI":"10.3109\/13693780802549594","article-title":"Biofilms of non-Candida albicans Candida species: Quantification, structure and matrix composition","volume":"47","author":"Silva","year":"2009","journal-title":"Med. Mycol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1494","DOI":"10.1038\/nprot.2008.141","article-title":"A simple and reproducible 96-well plate-based method for the formation of fungal biofilms and its application to antifungal susceptibility testing","volume":"3","author":"Pierce","year":"2008","journal-title":"Nat. Protoc."},{"key":"ref_34","unstructured":"Clinical and Laboratory Standards Institute (2017). Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts; CLSI standard M27, Clinical and Laboratory Standards Institute. [4th ed.]."},{"key":"ref_35","unstructured":"EUCAST-AFST (2020, April 22). Method for the determination of broth dilution minimum inhibitory concentrations of antifungal agents for yeasts\u2014Document E.DEF 7.3.2.. Available online: https:\/\/www.eucast.org\/fileadmin\/src\/media\/PDFs\/EUCAST_files\/AFST\/Files\/EUCAST_E_Def_7.3.2_Yeast_testing_definitive_revised_2020.pdf."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1759","DOI":"10.3389\/fmicb.2017.01759","article-title":"Tacrolimus Increases the Effectiveness of Itraconazole and Fluconazole against Sporothrix spp.","volume":"8","author":"Ramos","year":"2017","journal-title":"Front. Microbiol."}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/11\/2\/206\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:13:38Z","timestamp":1760134418000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/11\/2\/206"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,2,3]]},"references-count":36,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2022,2]]}},"alternative-id":["pathogens11020206"],"URL":"https:\/\/doi.org\/10.3390\/pathogens11020206","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,2,3]]}}}