Variations in virulence factors, antifungal susceptibility and extracellular polymeric substance compositions of cryptic and uncommon Candida species from oral candidiasis

Abstract Background Infections caused by uncommon Candida spp. (species other than C. albicans, C. parapsilosis, C. glabrata, C. tropicalis, and C. krusei) have been continuously reported worldwide. However, virulence factors and antifungal susceptibility profiles of uncommon Candida species remain...

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Main Authors: Orada Tosrisawatkasem, Sarut Thairat, Pairin Tonput, Pornpen Tantivitayakul
Format: Article
Language:English
Published: BMC 2025-07-01
Series:BMC Oral Health
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Online Access:https://doi.org/10.1186/s12903-025-06334-2
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author Orada Tosrisawatkasem
Sarut Thairat
Pairin Tonput
Pornpen Tantivitayakul
author_facet Orada Tosrisawatkasem
Sarut Thairat
Pairin Tonput
Pornpen Tantivitayakul
author_sort Orada Tosrisawatkasem
collection DOAJ
description Abstract Background Infections caused by uncommon Candida spp. (species other than C. albicans, C. parapsilosis, C. glabrata, C. tropicalis, and C. krusei) have been continuously reported worldwide. However, virulence factors and antifungal susceptibility profiles of uncommon Candida species remain limited. This study aimed to investigate the extracellular hydrolytic enzyme activities, antifungal susceptibility in planktonic and biofilm forms of cryptic and uncommon Candida species as well as their biofilm structures and extracellular polymeric substance (EPS) compositions. Methods Candida species was identified by sequence analysis of the internal transcribed spacer (ITS) regions of ribosomal RNA. The phospholipase and proteinase activities were evaluated by agar plate method. Biofilm-forming activity was analyzed using XTT-metabolic assay. Antifungal susceptibility tests for fluconazole, voriconazole, 5-flucytosine, and caspofungin were performed with broth microdilution following the CLSI guideline. Candida biofilms were stained with fluorescent dyes for fungal cells (SYTO9), extracellular proteins (SYPRO Ruby), exopolysaccharides (Alexa Fluor 635-conjugated Concanavalin A) and analyzed using confocal laser scanning microscopy (CLSM). Results A total of 25 uncommon Candida isolates including C. metapsilosis, C. orthopsilosis, C. guilliermondii (new nomenclature: Meyerozyma guilliermondii), C. fermentati (Meyerozyma caribbica), C. lusitaniae (Clavispora lusitaniae), C. rugosa (Diutina rugosa), C. pararugosa and C. nivariensis (Nakaseomyces nivariensis ) were identified, with 8% and 40% of the oral Candida isolates producing phospholipase and proteinase, respectively. All Candida isolates in this study were susceptible to fluconazole, voriconazole, and 5-flucytosine whereas 2 C. rugosa isolates (8% of all tested isolates) reduce susceptibility to caspofungin. The antifungal concentrations required to inhibit biofilm (sMIC) of C. rugosa and C. guilliermondii complex were higher than those of other species. In addition, the C. rugosa and C. guilliermondii biofilms had relatively high quantities of proteins, exopolysaccharides, and extracellular DNA (eDNA) compared to other species. Significant positive correlations between eDNA and exopolysaccharides (r s = 0.77, p < 0.01) and proteins (r s = 0.60, p < 0.01) in Candida biofilms were observed. Conclusion Our results indicated variation in the expression of hydrolytic enzyme production and biofilm forming activities among uncommon Candida spp. All Candida isolates exhibited susceptibility to azole and 5-flucytosine. This study also presented the visualization of biofilm structures and distribution of EPS components in Candida biofilms.
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spelling doaj-art-e8228159c1f443ca90adb66045b41cd52025-08-20T03:42:01ZengBMCBMC Oral Health1472-68312025-07-0125111210.1186/s12903-025-06334-2Variations in virulence factors, antifungal susceptibility and extracellular polymeric substance compositions of cryptic and uncommon Candida species from oral candidiasisOrada Tosrisawatkasem0Sarut Thairat1Pairin Tonput2Pornpen Tantivitayakul3Department of Oral Microbiology, Faculty of Dentistry, Mahidol UniversityResearch office, Faculty of Dentistry, Mahidol UniversityResearch office, Faculty of Dentistry, Mahidol UniversityDepartment of Oral Microbiology, Faculty of Dentistry, Mahidol UniversityAbstract Background Infections caused by uncommon Candida spp. (species other than C. albicans, C. parapsilosis, C. glabrata, C. tropicalis, and C. krusei) have been continuously reported worldwide. However, virulence factors and antifungal susceptibility profiles of uncommon Candida species remain limited. This study aimed to investigate the extracellular hydrolytic enzyme activities, antifungal susceptibility in planktonic and biofilm forms of cryptic and uncommon Candida species as well as their biofilm structures and extracellular polymeric substance (EPS) compositions. Methods Candida species was identified by sequence analysis of the internal transcribed spacer (ITS) regions of ribosomal RNA. The phospholipase and proteinase activities were evaluated by agar plate method. Biofilm-forming activity was analyzed using XTT-metabolic assay. Antifungal susceptibility tests for fluconazole, voriconazole, 5-flucytosine, and caspofungin were performed with broth microdilution following the CLSI guideline. Candida biofilms were stained with fluorescent dyes for fungal cells (SYTO9), extracellular proteins (SYPRO Ruby), exopolysaccharides (Alexa Fluor 635-conjugated Concanavalin A) and analyzed using confocal laser scanning microscopy (CLSM). Results A total of 25 uncommon Candida isolates including C. metapsilosis, C. orthopsilosis, C. guilliermondii (new nomenclature: Meyerozyma guilliermondii), C. fermentati (Meyerozyma caribbica), C. lusitaniae (Clavispora lusitaniae), C. rugosa (Diutina rugosa), C. pararugosa and C. nivariensis (Nakaseomyces nivariensis ) were identified, with 8% and 40% of the oral Candida isolates producing phospholipase and proteinase, respectively. All Candida isolates in this study were susceptible to fluconazole, voriconazole, and 5-flucytosine whereas 2 C. rugosa isolates (8% of all tested isolates) reduce susceptibility to caspofungin. The antifungal concentrations required to inhibit biofilm (sMIC) of C. rugosa and C. guilliermondii complex were higher than those of other species. In addition, the C. rugosa and C. guilliermondii biofilms had relatively high quantities of proteins, exopolysaccharides, and extracellular DNA (eDNA) compared to other species. Significant positive correlations between eDNA and exopolysaccharides (r s = 0.77, p < 0.01) and proteins (r s = 0.60, p < 0.01) in Candida biofilms were observed. Conclusion Our results indicated variation in the expression of hydrolytic enzyme production and biofilm forming activities among uncommon Candida spp. All Candida isolates exhibited susceptibility to azole and 5-flucytosine. This study also presented the visualization of biofilm structures and distribution of EPS components in Candida biofilms.https://doi.org/10.1186/s12903-025-06334-2Cryptic and uncommon CandidaVirulence factorsProteinasePhospholipaseBiofilm formationExtracellular polymeric substances
spellingShingle Orada Tosrisawatkasem
Sarut Thairat
Pairin Tonput
Pornpen Tantivitayakul
Variations in virulence factors, antifungal susceptibility and extracellular polymeric substance compositions of cryptic and uncommon Candida species from oral candidiasis
BMC Oral Health
Cryptic and uncommon Candida
Virulence factors
Proteinase
Phospholipase
Biofilm formation
Extracellular polymeric substances
title Variations in virulence factors, antifungal susceptibility and extracellular polymeric substance compositions of cryptic and uncommon Candida species from oral candidiasis
title_full Variations in virulence factors, antifungal susceptibility and extracellular polymeric substance compositions of cryptic and uncommon Candida species from oral candidiasis
title_fullStr Variations in virulence factors, antifungal susceptibility and extracellular polymeric substance compositions of cryptic and uncommon Candida species from oral candidiasis
title_full_unstemmed Variations in virulence factors, antifungal susceptibility and extracellular polymeric substance compositions of cryptic and uncommon Candida species from oral candidiasis
title_short Variations in virulence factors, antifungal susceptibility and extracellular polymeric substance compositions of cryptic and uncommon Candida species from oral candidiasis
title_sort variations in virulence factors antifungal susceptibility and extracellular polymeric substance compositions of cryptic and uncommon candida species from oral candidiasis
topic Cryptic and uncommon Candida
Virulence factors
Proteinase
Phospholipase
Biofilm formation
Extracellular polymeric substances
url https://doi.org/10.1186/s12903-025-06334-2
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