{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T23:28:24Z","timestamp":1788564504171,"version":"build-2803163510"},"reference-count":65,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2024,6,22]],"date-time":"2024-06-22T00:00:00Z","timestamp":1719014400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antibiotics"],"abstract":"<jats:p>Invasive candidiasis poses a worldwide threat because of the rising prevalence of antifungal resistance, resulting in higher rates of morbidity and mortality. Additionally, Candida species, which are opportunistic infections, have significant medical and economic consequences for immunocompromised individuals. This study explores the antifungal potential of chitosan to mitigate caspofungin resistance in caspofungin-resistant Candida albicans, C. krusei, and C. tropicalis isolates originating from human and animal sources using agar well diffusion, broth microdilution tests, and transmission electron microscope (TEM) analysis of treated Candida cells. Reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) was performed to assess the expression of SAGA complex genes (GCN5 and ADA2) and the caspofungin resistance gene (FKS) in Candida species isolates after chitosan treatment. The highest resistance rate was observed to ketoconazole (80%) followed by clotrimazole (62.7%), fluconazole (60%), terbinafine (58%), itraconazole (57%), miconazole (54.2%), amphotericin B (51.4%), voriconazole (34.28%), and caspofungin (25.7%). Nine unique FKS mutations were detected, including S645P (n = 3 isolates), S645F, L644F, S645Y, L688M, E663G, and F641S (one isolate in each). The caspofungin minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values before chitosan treatment ranged from 2 to 8 \u00b5g\/mL and 4 to 16 \u00b5g\/mL, respectively. However, the MIC and MFC values were decreased after chitosan treatment (0.0625\u20131 \u00b5g\/mL) and (0.125\u20132 \u00b5g\/mL), respectively. Caspofungin MIC was significantly decreased (p = 0.0007) threefold following chitosan treatment compared with the MIC values before treatment. TEM analysis revealed that 0.5% chitosan disrupted the integrity of the cell surface, causing irregular morphologies and obvious aberrant changes in cell wall thickness in caspofungin-resistant and sensitive Candida isolates. The cell wall thickness of untreated isolates was 0.145 \u03bcm in caspofungin-resistant isolate and 0.125 \u03bcm in sensitive isolate, while it was significantly lower in chitosan-treated isolates, ranging from 0.05 to 0.08 \u03bcm when compared with the cell wall thickness of sensitive isolate (0.03 to 0.06 \u03bcm). Moreover, RT-qPCR demonstrated a significant (p &lt; 0.05) decrease in the expression levels of histone acetyltransferase genes (GCN5 and ADA2) and FKS gene of caspofungin-resistant Candida species isolates treated with 0.5% chitosan when compared with before treatment (fold change values ranged from 0.001 to 0.0473 for GCN5, 1.028 to 4.856 for ADA2, and 2.713 to 12.38 for FKS gene). A comparison of the expression levels of cell wall-related genes (ADA2 and GCN5) between caspofungin-resistant and -sensitive isolates demonstrated a significant decrease following chitosan treatment (p &lt; 0.001). The antifungal potential of chitosan enhances the efficacy of caspofungin against various caspofungin-resistant Candida species isolates and prevents the development of further antifungal resistance. The results of this study contribute to the progress in repurposing caspofungin and inform a development strategy to enhance its efficacy, appropriate antifungal activity against Candida species, and mitigate resistance. Consequently, chitosan could be used in combination with caspofungin for the treatment of candidiasis.<\/jats:p>","DOI":"10.3390\/antibiotics13070578","type":"journal-article","created":{"date-parts":[[2024,6,24]],"date-time":"2024-06-24T08:42:03Z","timestamp":1719218523000},"page":"578","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Fighting Emerging Caspofungin-Resistant Candida Species: Mitigating Fks1-Mediated Resistance and Enhancing Caspofungin Efficacy by Chitosan"],"prefix":"10.3390","volume":"13","author":[{"given":"Aya","family":"Tarek","sequence":"first","affiliation":[{"name":"Faculty of Veterinary Medicine, Zagazig University, Zagazig 44511, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasmine H.","family":"Tartor","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Veterinary Medicine, Zagazig University, Zagazig 44511, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohamed N.","family":"Hassan","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Veterinary Medicine, Zagazig University, Zagazig 44511, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2157-5009","authenticated-orcid":false,"given":"Ioan","family":"Pet","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, Faculty of Bioengineering of Animal Resources, University of Life Sciences \u201cKing Mihai I\u201d from Timisoara, 300645 Timisoara, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8033-8463","authenticated-orcid":false,"given":"Mirela","family":"Ahmadi","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, Faculty of Bioengineering of Animal Resources, University of Life Sciences \u201cKing Mihai I\u201d from Timisoara, 300645 Timisoara, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adel","family":"Abdelkhalek","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Medicine, Badr University in Cairo (BUC), Badr City 11829, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,6,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"V\u00e9lez, N., Argel, A., Kissmann, A.K., Alp\u00edzar-Pedraza, D., Escand\u00f3n, P., Rosenau, F., St\u00e4ndker, L., and Firacative, C. (2024). Pore-forming peptide C14R exhibits potent antifungal activity against clinical isolates of Candida albicans and Candida auris. Front. Cell Infect. Microbiol., 14.","DOI":"10.3389\/fcimb.2024.1389020"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1017\/S0950268819001912","article-title":"Infectious disease outbreaks in the African region: Overview of events reported to the World Health Organization","volume":"147","author":"Mboussou","year":"2019","journal-title":"Epidemiol. Infect."},{"key":"ref_3","first-page":"1","article-title":"Spatial and temporal distribution of infectious disease epidemics, disasters and other potential public health emergencies in the World Health Organisation Africa region, 2016","volume":"16","author":"Okiro","year":"2020","journal-title":"Glob. Health"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"72","DOI":"10.3389\/fcimb.2022.807218","article-title":"Promising Anti-Biofilm Agents and Phagocytes Enhancers for the Treatment of Candida albicans Biofilm-Associated Infections","volume":"12","author":"Tartor","year":"2022","journal-title":"Front. Cell Infect. Microbiol."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Arastehfar, A., Gabald\u00f3n, T., Garcia-Rubio, R., Jenks, J.D., Hoenigl, M., Salzer, H.J.F., Ilkit, M., Lass-Fl\u00f6rl, C., and Perlin, D.S. (2020). Drug-Resistant Fungi: An Emerging Challenge Threatening Our Limited Antifungal Armamentarium. Antibiotics, 9.","DOI":"10.3390\/antibiotics9120877"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1016\/j.bcp.2016.11.019","article-title":"Antifungals","volume":"133","author":"Campoy","year":"2017","journal-title":"Biochem. Pharmacol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"876","DOI":"10.1080\/14756366.2022.2050224","article-title":"Echinocandins\u2014Structure, mechanism of action and use in antifungal therapy","volume":"37","author":"Chmielewska","year":"2022","journal-title":"J. Enzyme Inhib. Med. Chem."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"612","DOI":"10.1093\/cid\/civ791","article-title":"Echinocandin Resistance in Candida","volume":"61","author":"Perlin","year":"2015","journal-title":"Clin. Infect. Dis."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1111\/nyas.12831","article-title":"Mechanisms of echinocandin antifungal drug resistance","volume":"1354","author":"Perlin","year":"2015","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1374","DOI":"10.1038\/s41564-020-0760-7","article-title":"Epigenetic cell fate in Candida albicans is controlled by transcription factor condensates acting at super-enhancer-like elements","volume":"5","author":"Frazer","year":"2020","journal-title":"Nat. Microbiol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Mallick, E.M., Bergeron, A.C., Jones, S.K., Newman, Z.R., Brothers, K.M., Creton, R., Wheeler, R.T., and Bennett, R.J. (2016). Phenotypic Plasticity Regulates Candida albicans Interactions and Virulence in the Vertebrate Host. Front. Microbiol., 7.","DOI":"10.3389\/fmicb.2016.00780"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Conte, M., Eletto, D., Pannetta, M., Petrone, A.M., Monti, M.C., Cassiano, C., Giurato, G., Rizzo, F., Tessarz, P., and Petrella, A. (2022). Effects of Hst3p inhibition in Candida albicans: A genome-wide H3K56 acetylation analysis. Front. Cell Infect. Microbiol., 12.","DOI":"10.3389\/fcimb.2022.1031814"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Yu, S., Paderu, P., Lee, A., Eirekat, S., Healey, K., Chen, L., Perlin, D.S., and Zhao, Y. (2022). Histone Acetylation Regulator Gcn5 Mediates Drug Resistance and Virulence of Candida glabrata. Microbiol. Spectr., 10.","DOI":"10.1128\/spectrum.00963-22"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"6621","DOI":"10.1128\/MCB.19.10.6621","article-title":"The ADA complex is a distinct histone acetyltransferase complex in Saccharomyces cerevisiae","volume":"19","author":"Eberharter","year":"1999","journal-title":"Mol. Cell Biol."},{"key":"ref_15","first-page":"194614","article-title":"What do the structures of GCN5-containing complexes teach us about their function?","volume":"1864","author":"Helmlinger","year":"2021","journal-title":"BBA J."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Shih, P.Y., Liao, Y.T., Tseng, Y.K., Deng, F.S., and Lin, C.H. (2019). A Potential Antifungal Effect of Chitosan Against Candida albicans Is Mediated via the Inhibition of SAGA Complex Component Expression and the Subsequent Alteration of Cell Surface Integrity. Front. Microbiol., 10.","DOI":"10.3389\/fmicb.2019.00602"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Iracane, E., Vega-Est\u00e9vez, S., and Buscaino, A. (2021). On and Off: Epigenetic Regulation of C. albicans Morphological Switches. Pathogens, 10.","DOI":"10.3390\/pathogens10111463"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"3765","DOI":"10.1007\/s12010-021-03639-0","article-title":"Saccharomyces Cerevisiae as an Untapped Source of Fungal Chitosan for Antimicrobial Action","volume":"193","author":"Afroz","year":"2021","journal-title":"Appl. Biochem. Biotechnol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1128\/EC.4.4.703-715.2005","article-title":"Transcriptional response of Saccharomyces cerevisiae to the plasma membrane-perturbing compound chitosan","volume":"4","author":"Zakrzewska","year":"2005","journal-title":"Eukaryot. Cell"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"4862","DOI":"10.1128\/AAC.00027-12","article-title":"The presence of an FKS mutation rather than MIC is an independent risk factor for failure of echinocandin therapy among patients with invasive candidiasis due to Candida glabrata","volume":"56","author":"Shields","year":"2012","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Pogurschi, E.N., Petcu, C.D., Mizeranschi, A.E., Zugravu, C.A., Cirnatu, D., Pet, I., and Ghimpe\u021beanu, O.M. (2022). Knowledge, Attitudes and Practices Regarding Antibiotic Use and Antibiotic Resistance: A Latent Class Analysis of a Romanian Population. Int. J. Environ. Res. Public Health, 19.","DOI":"10.3390\/ijerph19127263"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"94832","DOI":"10.1371\/journal.pone.0094832","article-title":"Inhibition of Paracoccidioides lutzii Pb 01 isocitrate lyase by the natural compound argentilactone and its semi-synthetic derivatives","volume":"9","author":"Alves","year":"2014","journal-title":"PLoS ONE"},{"key":"ref_23","first-page":"97","article-title":"In vitro evaluation of antimicrobial activity of crude extracts of medicinal plants against multi drug resistant pathogens","volume":"2","author":"Rajendran","year":"2009","journal-title":"Biyol. Bilim. Araptyrma Derg."},{"key":"ref_24","first-page":"183","article-title":"Susceptibility of clinical Candida species isolates to antifungal agents by E-test, Southern Iran: A five-year study","volume":"3","author":"Badiee","year":"2011","journal-title":"Iran. J. Microbiol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Ben-Ami, R. (2018). Treatment of Invasive Candidiasis: A Narrative Review. J. Fungi, 4.","DOI":"10.3390\/jof4030097"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"761","DOI":"10.1007\/s15010-020-01475-8","article-title":"Emerging echinocandin-resistant Candida albicans and glabrata in Switzerland","volume":"48","author":"Coste","year":"2020","journal-title":"Infection"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"86","DOI":"10.3201\/eid1801.110556","article-title":"Candida spp. with acquired echinocandin resistance, France, 2004","volume":"18","author":"Dannaoui","year":"2012","journal-title":"Emerg. Infect. Dis."},{"key":"ref_28","first-page":"e361","article-title":"Evaluation of Antimicrobial and Antifungal efficacy of Chitosan as endodontic irrigant against Enterococcus faecalis and Candida albicans Biofilm formed on tooth substrate","volume":"9","author":"Yadav","year":"2017","journal-title":"J. Clin. Exp. Dent."},{"key":"ref_29","first-page":"1","article-title":"Nanoparticles and irradiated chitosan impact on the quality of sweet green bell pepper (Capsicum annuum L.) under cold storage conditions","volume":"30","author":"Ewais","year":"2022","journal-title":"Arab J. Nucl. Sci. Appl."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Lo, W.H., Deng, F.S., Chang, C.J., and Lin, C.H. (2020). Synergistic Antifungal Activity of Chitosan with Fluconazole against Candida albicans, Candida tropicalis, and Fluconazole-Resistant Strains. Molecules, 25.","DOI":"10.3390\/molecules25215114"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"5156","DOI":"10.3390\/md13085156","article-title":"Chitosan: An update on potential biomedical and pharmaceutical applications","volume":"13","author":"Cheung","year":"2015","journal-title":"Mar. Drugs"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/j.ijfoodmicro.2010.09.012","article-title":"Antimicrobial properties of chitosan and mode of action: A state of the art review","volume":"144","author":"Kong","year":"2010","journal-title":"Int. J. Food Microbiol."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Poznanski, P., Hameed, A., and Orczyk, W. (2023). Chitosan and chitosan nanoparticles: Parameters enhancing antifungal activity. Molecules, 28.","DOI":"10.3390\/molecules28072996"},{"key":"ref_34","first-page":"1457","article-title":"Chitosan as antimicrobial agent: Applications and mode of action","volume":"4","author":"Rabea","year":"2003","journal-title":"J. Biol. Macromol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1686","DOI":"10.1016\/j.ijbiomac.2020.09.200","article-title":"The antibacterial structure-activity relationship for common chitosan derivatives","volume":"165","author":"Rathinam","year":"2020","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1016\/j.ijfoodmicro.2014.04.029","article-title":"Influence of acetylation degree and molecular weight of homogeneous chitosans on antibacterial and antifungal activities","volume":"185","author":"Younes","year":"2014","journal-title":"Int. J. Food Microbiol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"662","DOI":"10.1016\/j.carbpol.2018.04.091","article-title":"Effect of the molecular weight of chitosan on its antifungal activity against Candida spp. in planktonic cells and biofilm","volume":"195","author":"Garcia","year":"2018","journal-title":"Carbohydr. Polym."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1016\/j.ijbiomac.2019.12.031","article-title":"Antifungal activity of different molecular weight chitosans against planktonic cells and biofilm of Sporothrix brasiliensis","volume":"143","author":"Garcia","year":"2020","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"818","DOI":"10.1016\/j.foodcont.2015.07.011","article-title":"Effect of chitosan coating combined with pomegranate peel extract on the quality of Pacific white shrimp during iced storage","volume":"59","author":"Yuan","year":"2016","journal-title":"Food Control."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"239","DOI":"10.4315\/0362-028X-62.3.239","article-title":"Antibacterial Activity of Shrimp Chitosan against Escherichia coli","volume":"62","author":"Tsai","year":"1999","journal-title":"J. Food Prot."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"566","DOI":"10.1016\/j.carbpol.2008.04.010","article-title":"Antimicrobial activity of piperazine derivatives of chitosan","volume":"74","author":"Holappa","year":"2008","journal-title":"Carbohydr. Polym."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Ke, C.L., Deng, F.S., Chuang, C.Y., and Lin, C.H. (2021). Antimicrobial actions and applications of chitosan. Polymers, 13.","DOI":"10.3390\/polym13060904"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"855","DOI":"10.1128\/EC.00399-06","article-title":"Chitosan, the deacetylated form of chitin, is necessary for cell wall integrity in Cryptococcus neoformans","volume":"6","author":"Baker","year":"2007","journal-title":"Eukaryot. Cell"},{"key":"ref_44","first-page":"5","article-title":"Effects of chitosan on Candida albicans: Conditions for its antifungal activity","volume":"2013","author":"Calahorra","year":"2013","journal-title":"Biomed. Res. Int."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1005","DOI":"10.1080\/21505594.2022.2081291","article-title":"Biomarkers of caspofungin resistance in Candida albicans isolates: A proteomic approach","volume":"13","author":"Hafez","year":"2022","journal-title":"Virulence"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1059","DOI":"10.1002\/jobm.201500775","article-title":"Cell wall composition plays a key role on sensitivity of filamentous fungi to chitosan","volume":"56","year":"2016","journal-title":"J. Basic. Microbiol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1016\/j.carbpol.2017.02.001","article-title":"Recent developments in antibacterial and antifungal chitosan and its derivatives","volume":"164","author":"Verlee","year":"2017","journal-title":"Carbohydr. Polym."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1146\/annurev.biochem.70.1.475","article-title":"Transcriptional coactivator complexes","volume":"70","author":"Lemon","year":"2001","journal-title":"Annu. Rev. Biochem."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"2389","DOI":"10.1091\/mbc.e08-11-1093","article-title":"Genome-wide mapping of the coactivator AdA2p yields insight into the functional roles of SAGA\/ADA complex in Candida albicans","volume":"20","author":"Sellam","year":"2009","journal-title":"J. Mol. Cell Biol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"4807","DOI":"10.1002\/j.1460-2075.1994.tb06806.x","article-title":"Functional similarity and physical association between GCN5 and ADA2: Putative transcriptional adaptors","volume":"13","author":"Marcus","year":"1994","journal-title":"EMBO J."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Yien, L., Zin, N.M., Sarwar, A., and Katas, H. (2012). Antifungal activity of chitosan nanoparticles and correlation with their physical properties. Int. J. Biomater., 632698.","DOI":"10.1155\/2012\/632698"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"9445","DOI":"10.1038\/s41598-019-45817-5","article-title":"The fungal histone acetyl transferase Gcn5 controls virulence of the human pathogen Candida albicans through multiple pathways","volume":"9","author":"Shivarathri","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1359","DOI":"10.1111\/j.1469-0691.2010.03398.x","article-title":"Improved clinical laboratory identification of human pathogenic yeasts by matrix-assisted laser desorption ionization time-of-flight mass spectrometry","volume":"17","author":"Bader","year":"2011","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_54","unstructured":"CLSI (2018). Method for Antifungal Disk Diffusion Susceptibility Testing of Yeasts, (M44-ED3), Clinical and Laboratory Standards Institute. [3rd ed.]."},{"key":"ref_55","unstructured":"CLSI (2022). Performance Standards for Antifungal Susceptibility Testing of Yeasts, Clinical and Laboratory Standards Institute. [3rd ed.]."},{"key":"ref_56","unstructured":"NCCLS (2002). Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts, NCCLS. [2nd ed.]."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"3783","DOI":"10.1128\/AAC.00473-08","article-title":"Development of caspofungin resistance following prolonged therapy for invasive candidiasis secondary to Candida glabrata infection","volume":"52","author":"Thompson","year":"2008","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_58","first-page":"Dc01-4","article-title":"Comparative Analysis of Disc Diffusion and E-test with Broth Micro-dilution for Susceptibility Testing of Clinical Candida Isolates Against Amphotericin B, Fluconazole, Voriconazole and Caspofungin","volume":"9","author":"Kumar","year":"2015","journal-title":"J. Clin. Diagn. Res."},{"key":"ref_59","doi-asserted-by":"crossref","unstructured":"Vazquez-Mu\u00f1oz, R., Avalos-Borja, M., and Castro-Longoria, E. (2014). Ultrastructural analysis of Candida albicans when exposed to silver nanoparticles. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0108876"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"628","DOI":"10.1093\/mmy\/myw015","article-title":"The conserved dual phosphorylation sites of the Candida albicans Hog1 protein are crucial for white\u2013opaque switching, mating, and pheromone-stimulated cell adhesion","volume":"54","author":"Chang","year":"2016","journal-title":"Sabouraudia"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"245","DOI":"10.5395\/rde.2012.37.4.245","article-title":"Statistical notes for clinical researchers: Assessing normal distribution (1)","volume":"37","author":"Kim","year":"2012","journal-title":"Restor. Dent. Endod."},{"key":"ref_63","unstructured":"Cary, N. (2012). SAS\/STAT statistics user\u2019s guide. Statistical Analytical System, SAS Institute Inc.. [5th ed.]."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"3092","DOI":"10.1128\/AAC.00088-08","article-title":"Mutations in the fks1 gene in Candida albicans, C. tropicalis, and C. krusei correlate with elevated caspofungin MICs uncovered in AM3 medium using the method of the European Committee on Antibiotic Susceptibility Testing","volume":"52","author":"Bretagne","year":"2008","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1128\/AAC.01926-17","article-title":"Development of Echinocandin Resistance in Candida tropicalis following Short-Term Exposure to Caspofungin for Empiric Therapy","volume":"62","author":"Khan","year":"2018","journal-title":"Antimicrob. Agents Chemother."}],"container-title":["Antibiotics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-6382\/13\/7\/578\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T15:03:00Z","timestamp":1760108580000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-6382\/13\/7\/578"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,6,22]]},"references-count":65,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2024,7]]}},"alternative-id":["antibiotics13070578"],"URL":"https:\/\/doi.org\/10.3390\/antibiotics13070578","relation":{},"ISSN":["2079-6382"],"issn-type":[{"value":"2079-6382","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,6,22]]}}}