{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T06:52:19Z","timestamp":1788245539453,"version":"build-2803163510"},"reference-count":42,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2020,8,20]],"date-time":"2020-08-20T00:00:00Z","timestamp":1597881600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>The growing number of oral infections caused by the Candida species are becoming harder to treat as the commonly used antibiotics become less effective. This drawback has led to the search for alternative strategies of treatment, which include the use of antifungal molecules derived from natural products. Herein, crotoxin (CTX), the main toxin of Crotalus durissus terrificus venom, was challenged against Candida tropicalis (CBS94) and Candida dubliniensis (CBS7987) strains by in vitro antimicrobial susceptibility tests. Minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC), and inhibition of biofilm formation were evaluated after CTX treatment. In addition, CTX-induced cytotoxicity in HaCaT cells was assessed by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) colorimetric assay. Native CTX showed a higher antimicrobial activity (MIC = 47 \u03bcg\/mL) when compared to CTX-containing mouthwash (MIC = 750 \u03bcg\/mL) and nystatin (MIC = 375 \u03bcg\/mL). Candida spp biofilm formation was more sensitive to both CTX and CTX-containing mouthwash (IC100 = 12 \u03bcg\/mL) when compared to nystatin (IC100 &gt; 47 \u03bcg\/mL). Moreover, significant membrane permeabilization at concentrations of 1.5 and 47 \u00b5g\/mL was observed. Native CTX was less cytotoxic to HaCaT cells than CTX-containing mouthwash or nystatin between 24 and 48 h. These preliminary findings highlight the potential use of CTX in the treatment of oral candidiasis caused by resistant strains.<\/jats:p>","DOI":"10.3390\/toxins12090532","type":"journal-article","created":{"date-parts":[[2020,8,20]],"date-time":"2020-08-20T21:13:42Z","timestamp":1597958022000},"page":"532","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Evaluation of the Effectiveness of Crotoxin as an Antiseptic against Candida spp. Biofilms"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9759-6060","authenticated-orcid":false,"given":"Amanda Pissinatti","family":"Canelli","sequence":"first","affiliation":[{"name":"Graduate Program of Biomedical Sciences, University Center of Herm\u00ednio Ometto Foundation\u2013FHO, Araras, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6327-8443","authenticated-orcid":false,"given":"Ta\u00eds Fernanda","family":"dos Santos Rodrigues","sequence":"additional","affiliation":[{"name":"Graduate Program of Biomedical Sciences, University Center of Herm\u00ednio Ometto Foundation\u2013FHO, Araras, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vivian Fernandes Furletti","family":"de Goes","sequence":"additional","affiliation":[{"name":"Graduate Program of Biomedical Sciences, University Center of Herm\u00ednio Ometto Foundation\u2013FHO, Araras, SP, Brazil"},{"name":"Graduate Program of Orthodontics, University Center of Herm\u00ednio Ometto Foundation\u2013FHO, Araras, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4418-1080","authenticated-orcid":false,"given":"Guilherme Ferreira","family":"Caetano","sequence":"additional","affiliation":[{"name":"Graduate Program of Biomedical Sciences, University Center of Herm\u00ednio Ometto Foundation\u2013FHO, Araras, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7553-5219","authenticated-orcid":false,"given":"Maur\u00edcio Ventura","family":"Mazzi","sequence":"additional","affiliation":[{"name":"Graduate Program of Biomedical Sciences, University Center of Herm\u00ednio Ometto Foundation\u2013FHO, Araras, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,8,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1016\/j.toxicon.2012.05.009","article-title":"Crystallographic characterization of functional sites of crotoxin and ammodytoxin, potent \u03b2-neurotoxins from Viperidae venom","volume":"60","author":"Faure","year":"2012","journal-title":"Toxicon"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1045","DOI":"10.1016\/j.toxicon.2010.01.011","article-title":"Crotoxin: Novel activities for a classic beta-neurotoxin","volume":"55","author":"Sampaio","year":"2010","journal-title":"Toxicon"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1016\/j.bbi.2019.12.009","article-title":"Crotoxin down-modulates pro-inflammatory cells and alleviates pain on the MOG35-55-induced experimental autoimmune encephalomyelitis, an animal model of multiple sclerosis","volume":"84","author":"Teixeira","year":"2020","journal-title":"Brain Behav. Immun."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"502","DOI":"10.1177\/0310057X0803600404","article-title":"Chlorhexidine\u2014Pharmacology and clinical applications","volume":"36","author":"Lim","year":"2008","journal-title":"Anaesth. Intensive Care"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Fiorillo, L. (2019). Chlorhexidine gel use in the oral district: A systematic review. Gels, 5.","DOI":"10.3390\/gels5020031"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"376","DOI":"10.1111\/apm.12688","article-title":"Dental biofilm infections\u2014An update","volume":"125","author":"Larsen","year":"2017","journal-title":"Apmis"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"442","DOI":"10.1111\/j.1567-1364.2007.00347.x","article-title":"Characteristics of biofilm formation by Candida tropicalis and antifungal resistance","volume":"8","author":"Bizerra","year":"2008","journal-title":"FEMS Yeast Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1128\/CMR.00013-11","article-title":"Polymicrobial interactions: Impact on pathogenesis and human disease","volume":"25","author":"Peters","year":"2012","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Allison, D.L., Willems, H.M.E., Jayatilake, J.A.M.S., Bruno, V.M., Peters, B.M., and Shirtliff, M.E. (2016). Candida-Bacteria Interactions: Their Impact on Human Disease. Microbiol. Spectr., 4.","DOI":"10.1128\/microbiolspec.VMBF-0030-2016"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"518","DOI":"10.1111\/odi.12691","article-title":"Is the oral fungal pathogen Candida albicans a cariogen?","volume":"24","author":"Pereira","year":"2018","journal-title":"Oral Dis."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1002\/1521-4028(200206)42:3<207::AID-JOBM207>3.0.CO;2-C","article-title":"Candida dubliniensis, a new fungal pathogen","volume":"42","author":"Morales","year":"2002","journal-title":"J. Basic Microbiol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"959","DOI":"10.1099\/jmm.0.008391-0","article-title":"Isolation of Candida dubliniensis from denture wearers","volume":"58","author":"Gasparoto","year":"2009","journal-title":"J. Med. Microbiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1111\/j.1741-2358.2010.00407.x","article-title":"Prevalence and antifungal drug sensitivity of non-albicans Candida in oral rinse samples of self-caring elderly","volume":"28","author":"Meurman","year":"2010","journal-title":"Gerodontology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"208","DOI":"10.3109\/13693786.2010.507605","article-title":"Assessment of Candida species colonization and denture-related stomatitis in complete denture wearers","volume":"4","author":"Zomorodian","year":"2011","journal-title":"Med. Mycol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"873","DOI":"10.1099\/jmm.0.013227-0","article-title":"Candida tropicalis: Its prevalence, pathogenicity and increasing resistance to fluconazole","volume":"59","author":"Kothavade","year":"2010","journal-title":"J. Med. Microbiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2554","DOI":"10.2174\/1568026619666191025153124","article-title":"The Chemistry of Drugs to Treat Candida albicans","volume":"9","author":"Ortiz","year":"2019","journal-title":"Curr. Top. Med. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1119","DOI":"10.2174\/1568026616666160930151242","article-title":"Animal Venom Peptides: Potential for new antimicrobial agents","volume":"17","author":"Macedo","year":"2017","journal-title":"Curr. Top. Med. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1002\/ddr.21456","article-title":"Harnessing snake venom phospholipases A2 to novel approaches for overcoming antibiotic resistance","volume":"80","author":"Almeida","year":"2019","journal-title":"Drug Dev. Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"14760","DOI":"10.1021\/jp2062496","article-title":"Protein-surfactant interaction: Sodium dodecyl sulfate-induced unfolding of ribonuclease A","volume":"115","author":"Naidu","year":"2011","journal-title":"J. Phys. Chem. B"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"665","DOI":"10.1089\/omi.2010.0041","article-title":"Phospholipidome of Candida: Each species of Candida has distinctive phospholipid molecular species","volume":"14","author":"Singh","year":"2010","journal-title":"OMICS J. Integr. Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"131","DOI":"10.3314\/jjmm.51.131","article-title":"Cell surface hydrophobicity and adhesion: A study on fifty clinical isolates of Candida albicans","volume":"51","author":"Raut","year":"2010","journal-title":"Nihon Ishinkin Gakkai Zasshi"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1080\/20477724.2018.1442764","article-title":"Targeting the GPI biosynthetic pathway","volume":"112","author":"Yadav","year":"2018","journal-title":"Pathog. Glob. Health"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"631","DOI":"10.5433\/1679-0359.2008v29n3p631","article-title":"Beta-glucana from Saccharomyces cerevisiae: Constitution, bioactivity and obtaining","volume":"29","author":"Magnani","year":"2008","journal-title":"Semin. Ci\u00eanc. Agr\u00e1r."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"8074","DOI":"10.1021\/bi00246a028","article-title":"Multiplicity of acidic subunit isoforms of crotoxin, the phospholipase A2 neurotoxin from Crotalus durissus terrificus venom, results from posttranslational modifications","volume":"30","author":"Faure","year":"1991","journal-title":"Biochemistry"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1007\/BF00873958","article-title":"In vitro comparison of cytotoxic effects of crotoxin against three human tumors and a normal human epidermal keratinocyte cell line","volume":"12","author":"Rudd","year":"1994","journal-title":"Investig. New Drugs"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1111\/eos.12177","article-title":"The in vitro impact of toothpaste extracts on cell viability","volume":"123","author":"Cvikl","year":"2015","journal-title":"Eur. J. Oral Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/S0041-0101(96)00061-X","article-title":"Encapsulation of native crotoxin in liposomes: A safe approach for the production of antivenom and vaccination against Crotalus durissus terrificus venom","volume":"35","author":"Freitas","year":"1997","journal-title":"Toxicon"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1042\/BA20000080","article-title":"Effect of membrane composition and of co-encapsulation of immunostimulants in a liposome-entrapped crotoxin","volume":"33","year":"2001","journal-title":"Biotechnol. Appl. Biochem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"510","DOI":"10.1021\/mp900232a","article-title":"Studies on the mechanism of action of dextrin\u2212phospholipase A2 and its suitability for use in combination therapy","volume":"7","author":"Ferguson","year":"2010","journal-title":"Mol. Pharm."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"722","DOI":"10.1271\/bbb.66.722","article-title":"Coupling reaction and properties of poly(ethylene glycol)-linked phospholipases A2","volume":"66","author":"Bianco","year":"2002","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1397","DOI":"10.1038\/aps.2011.116","article-title":"Anti-tumor activity of CrTX in human lung adenocarcinoma cell line A549","volume":"32","author":"Ye","year":"2011","journal-title":"Acta Pharmacol. Sin."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1177\/1073858405285923","article-title":"Phospholipase A2-generated lipid mediators in the brain: The good, the bad, and the ugly","volume":"12","author":"Farooqui","year":"2006","journal-title":"Neuroscientist"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1016\/j.alit.2014.07.005","article-title":"Secreted phospholipase A2 and mast cells","volume":"64","author":"Murakami","year":"2015","journal-title":"Allergol. Int."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1186\/s40409-018-0178-3","article-title":"Immunotherapeutic potential of Crotoxin: Anti-inflammatory and immunosuppressive properties","volume":"24","author":"Sartim","year":"2018","journal-title":"J. Venom. Anim. Toxins Trop. Dis."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"7435621","DOI":"10.1155\/2017\/7435621","article-title":"HaCaT cells as a reliable in vitro differentiation model to dissect the inflammatory\/repair response of human keratinocytes","volume":"2017","author":"Colombo","year":"2017","journal-title":"Mediat. Inflamm."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.toxicon.2018.10.306","article-title":"Crotoxin from Crotalus durissus terrificus venom: In vitro cytotoxic activity of a heterodimeric phospholipase A2 on human cancer-derived cell lines","volume":"156","author":"Muller","year":"2018","journal-title":"Toxicon"},{"key":"ref_37","unstructured":"CLSI (2017). Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts, Clinical and Laboratory Standards Institute. [4th ed.]. CLSI standard M27."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"7495218","DOI":"10.1155\/2018\/7495218","article-title":"Antifungal Susceptibility and Biofilm Production of Candida spp. Isolated from Clinical Samples","volume":"2018","author":"Marak","year":"2018","journal-title":"Int. J. Microbiol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"96","DOI":"10.2174\/1874285801105010096","article-title":"Inhibition of growth of highly resistant bacterial and fungal pathogens by a natural product","volume":"5","author":"Hafidh","year":"2011","journal-title":"Open Microbiol. J."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1128\/AAC.01599-12","article-title":"Efficacy of ethanol against Candida albicans and Staphylococcus aureus polymicrobial biofilms","volume":"57","author":"Peters","year":"2013","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1016\/j.micres.2016.12.008","article-title":"Antifungal activity of ionic liquids based on (-)-menthol: A mechanism study","volume":"197","author":"Suchodolski","year":"2017","journal-title":"Microbiol. Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1016\/0022-1759(86)90368-6","article-title":"Rapid colorimetric assay for cell growth and survival: Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability","volume":"89","author":"Denizot","year":"1986","journal-title":"J. Immunol. Methods"}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/12\/9\/532\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:03:38Z","timestamp":1760177018000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/12\/9\/532"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,8,20]]},"references-count":42,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2020,9]]}},"alternative-id":["toxins12090532"],"URL":"https:\/\/doi.org\/10.3390\/toxins12090532","relation":{},"ISSN":["2072-6651"],"issn-type":[{"value":"2072-6651","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,8,20]]}}}