Research Article
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Investigation of Antifungal Susceptibility of Candida Species Isolated from Various Clinical Samples with Disc Diffusion Method

Year 2025, Volume: 15 Issue: 3, 473 - 478, 24.09.2025
https://doi.org/10.33631/sabd.1747296

Abstract

Aim: Increasing frequency of infections due to Candida species in recent years and widespread use of empirical antifungal cause the development of resistant Candida strains and increased resistance rates. Therefore, the need for in vitro antifungal susceptibility tests is increasing in selecting appropriate and effective antifungal treatments.
Material and Methods: In this study, Candida strains isolated from various clinical specimens from Düzce University Clinical Microbiology Laboratory were typed and their susceptibility to antifungal agents were investigated. Identification of origins were made using conventional and auotomatic methods. Antifungal susceptibilities of strains were determined by disc diffusion method. The interpretation of zone diameters was performed according to the CLSI M44-S3 document for voriconazole and fluconazole, and according to the recommendations of Rosco Diagnostica, the manufacturer of Neo-Sensitabs tablets, for posaconazole, caspofungin, and itraconazole. No threshold values have been established for amphotericinB and flucytosine.
Results: According to this study 156 Candida strains were found as 89 (57.1%) C. albicans, 48 (30.8%) C. glabrata, 11 (7.1%) C. tropicalis, 2(1.5%) C. lusitaniae, 2 (1.3%) C. kefyr, 2 (1.3%) C. parapsilosis and 2 (1.3%) C. guilliermondii.
When the antifungal susceptibilities of strains were examined, antifungals with the highest resistance were detected as itraconazole (64.1%); the antifungals with the highest susceptibilities were as caspofungin (98.7%), voriconazole (94.2%), fluconazole (85.3%) and the highest dose dependent susceptibilities were determined as itraconazole (34.6%) and posacanozole (11.5%) respectively. The mean zone diameters for amphotericin B and flucytosine were measured as 15.29 (± 3), 17.06 (± 9.8) respectively.
Conclusion: Identification of agents for proper treatment of Candida species and testing of antifungal susceptibility tests of isolated strains will contribute to empirical antifungal treatment policies.

Project Number

2015.04.01.353

References

  • Siddiqui R, Mendiratta DK, Siddiqui AF, Rukadikar A. A study of the association between virulence factors and antifungal susceptibility profile of Candida species recovered from cases of vulvovaginal candidiasis. J Family Med Prim Care. 2023;12(1):152-9.
  • R AN, Rafiq NB. Candidiasis. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2025 [cited 2025 May 02]. Available from: https://www.ncbi.nlm.nih.gov/books/NBK560624/.
  • Achkar JM, Fries BC. Candida infections of the Genitourinary Tract. Clin Microbiol Rev. 2010;23(2):253-73.
  • Lamoth F, Lockhart SR, Berkow EL, Calandra T. Changes in the epidemiological landscape of invasive candidiasis. J Antimicrob Chemother. 2018;73:4-13.
  • El-Kholy MA, Helaly GF, El Ghazzawi EF, El-Sawaf G, Shawky SM. Virulence factors and antifungal susceptibility profile of C. tropicalis isolated from various clinical specimens in Alexandria, Egypt. J Fungi. 2021;7(5):351.
  • Makled AF, Ali SAM, Labeeb AZ, Salman SS, Shebl DZM, Hegazy SG, et al. Characterization of Candida species isolated from clinical specimens: insights into virulence traits, antifungal resistance and molecular profiles. BMC Microbiol. 2024;24(1):388.
  • Clinical and Laboratory Standards Institute (CLSI). Method for antifungal disk diffusion susceptibility testing of yeasts. Approved guideline-second edition, 2nd ed. CLSI document M44-A2. Wayne (PA): CLSI; 2009.
  • Jayachandran AL, Katragadda R, Ravinder T, Vajravelu L, Manorajan L, Hemalatha S, et al. Antifungal susceptibility pattern among Candida species: an evaluation of disc diffusion and micro broth dilution method. J Microbil Infect Dis. 2018;08(03):97-101.
  • Larone DH. Yeasts and yeast like organisms. In: Larone DH. editor. Medically important fungi: a guide to identification. 5th ed. Washington (DC): ASM Press; 2011. p. 117-54.
  • Espinel-Ingroff A, Canton E. Comparison of Neo-Sensitabs tablet diffusion assay with CLSI broth microdilution M38-A and disk diffusion methods for testing susceptibility of filamentous fungi with amphotericin B, caspofungin, itraconazole, posaconazole, and voriconazole. J Clin Microbiol. 2008;46(5):1793-803.
  • Marr KA, Seidel K, White TC, Bowden RA. Candidemia in allogeneic blood and marrow transplant recipients: evolution of risk factors after the adoption of prophylactic fluconazole. J Infect Dis. 2000;181(1):309-16.
  • Ener B. İnvitro antifungal duyarlılık testleri: standardizasyon ve klinik önemi. Mikrobiyol Bül. 1996;30:419-25.
  • Chen X, Shen M, Feng Q, Yang G, Tian R, Yao S, et al. Epidemiological characteristics, antifungal susceptibility, and mortality factors of candidemia in adults at a tertiary teaching hospital in Zunyi, China (2016-2023). BMC Infect Dis. 2025;25(1):726.
  • Zhang W, Song X, Wu H, Zheng R. Epidemiology, risk factors and outcomes of Candida albicans vs. non-albicans candidaemia in adult patients in Northeast China. Epidemiol Infect. 2019;25:147:e277.
  • Bayram Y, Gültepe B, Özlük S, Güdücüoğlu H. Çeşitli klinik örneklerden izole edilen Candida kökenlerinin identifikasyonu ve antifungal duyarlılıklarının araştırılması. Van Tıp Dergisi. 2012;19(4):177-81.
  • Sav H, Demir G, Atalay MA, Koç AN. Klinik örneklerden izole edilen Candida türlerinin değerlendirilmesi. Turk Hij Den Biyol Derg. 2013;70(4):175-80.
  • Pham CD, Iqbal N, Bolden CB, Kuykendall RJ, Harrison LH, Farley MM, et al. Role of FKS Mutations in Candida glabrata: MIC values, echinocandin resistance, and multidrug resistance. Antimicrob Agents Chemother. 2014;58(8):4690-6.
  • Daneshnia F, de Almeida Júnior JN, Ilkit M, Lombardi L, Perry AM, Gao M, et al. Worldwide emergence of fluconazole-resistant Candida parapsilosis: current framework and future research roadmap. Lancet Microbe. 2023;4(6):e470-80.
  • Özkaya E, Çalışkan A, Kirişci Ö, Tüme S. Son üç yılda Kahramanmaraş Necip Fazıl Şehir Hastanesinde kan kültürlerinden izole edilen Candida türlerinin dağılımı ve antifungal duyarlılıkları. Türk Mikrobiyol Cem Derg. 2016;46(2):63-8.
  • Diekema DJ, Messer SA, Hollis RJ, Boyken LB, Tendolkar S, Kroeger J, et al. Evaluation of Etest and disk diffusion methods compared with broth microdilution antifungal susceptibility testing of clinical isolates of Candida spp. against posaconazole. J Clin Microbiol. 2007;45(6):1974-7.
  • Jaruvongvanich V, Worasilchai N, Kongnatthasate K, Jaruvongvanich S, Damkerngsuntorn W, Atikarnbodee D, et al. Correlation between broth microdilution, E-test and disk diffusion methods for testing antifungal susceptibility of Candida species isolated from Thai blood samples. Asian Biomed. 2016;10(1):75-80.
  • Giri S, Kindo AJ. Evaluation of antifungal susceptibility testing in Candida isolates by Candifast and disk-diffusion method. Indian J Pathol Microbiol. 2014;57(4):595-7.
  • Altuncu E, Bilgen H, Çerikcioğlu N, İlki A, Ülger N, Bakır M, et al. Neonatal Candida infeksiyonları ve etkenlerinin antifungal duyarlılıkları. Mikrobiyol Bul. 2010;44:593-603.
  • Yiğit N, Aktaş E. Activities of amphotericin B, fluconazole and voriconazole against Candida bloodstream isolates determined by broth microdilution and disk diffusion methods. Turk Hij Den Biyol Derg. 2014;71(3):131-40.
  • Kubiak DW, Farmakiotis D, Arons V, Hollins RM, Rostas SE, Weiser LM, et al. Utility of in-house fluconazole disk diffusion susceptibility testing in thetreatment of candidemia. Diagn Microbiol Infect Dis. 2016;84(3):223-6.
  • Azevedo AC, Bizerra FC, da Matta DA, de Almeida LP, Rosas R, Colombo AL. In vitro susceptibility of a large collection of Candida strains against fluconazole and voriconazole by using the CLSI disk diffusion assay. Mycopathologia. 2011;171(6):411-6.
  • Pathak AK, Jain NR, Joshi R. Antibiogram of Candida species isolated from mono and multi‑species oral candidal carriage using disk diffusion method. Saudi J Health Sci. 2012;1(3):132-8.

Çeşitli Klinik Örneklerden İzole Edilen Candida Türlerinin Antifungal Duyarlılıklarının Disk Difüzyon Yöntemiyle Araştırılması

Year 2025, Volume: 15 Issue: 3, 473 - 478, 24.09.2025
https://doi.org/10.33631/sabd.1747296

Abstract

Amaç: Son yıllarda Candida türlerine bağlı infeksiyonların sıklığının giderek artması ve ampirik antifungal kullanımının yaygınlaşması, dirençli Candida suşlarının ortaya çıkmasına ve direnç oranlarının artmasına neden olmaktadır. Bu nedenle uygun ve etkin antifungal tedavinin seçiminde in vitro antifungal duyarlılık testlerine gereksinim artmaktadır.
Gereç ve Yöntemler: Bu çalışmada, Düzce Üniversitesi Tıbbi Mikrobiyoloji Laboratuvarı’na gelen çeşitli klinik örneklerden izole edilen Candida suşları tiplendirilerek antifungal ajanlara karşı duyarlılıkları araştırıldı. Kökenlerin tanımlanması konvansiyonel ve otomatize yöntemler kullanılarak yapıldı. Suşların antifungal duyarlılıkları Kirby-bauer disk difüzyon yöntemi ile yapıldı. Zon çaplarının yorumlanması, vorikonazol ve flukonazol için CLSI M44-S3 dokümanına göre, posakonazol, kaspofungin ve itrakonazol için ise NeoSensitabs tabletlerinin üretici firması olan Rosco Diagnostica’nın önerilerine göre yapılmıştır. Amfoterisin B ve flusitozin için ise kabul edilmiş sınır değerler literatürde saptanamamıştır.
Bulgular: Çalışmaya dahil edilen 156 Candida suşunun 89’u (%57,1) C. albicans, 48’i (%30,8) C. glabrata, 11’i (%7,1) C. tropicalis, 2’si (%1,5) C. lusitaniae, 2’si (%1,3) C. kefyr, 2’si (%1,3) C. parapsilosis ve 2’si (%1,3) C. guilliermondii olarak saptanmıştır. Suşların antifungal duyarlılıkları incelendiğinde en fazla direnç saptanan antifungal itrakonazol (%64,1); en fazla duyarlılık saptanan antifungaller sırasıyla kaspofungin (%98,7), vorikonazol (%94,2), flukonazol (%85,3) en fazla doza bağlı duyarlılık saptanan antifungaller ise sırasıyla itrakonazol (%34,6) ve posakanozol (%11,5) olarak saptanmıştır. Amfoterisin B ve flusitozin için belirlenen ortalama zon çapları sırasıyla 15,29 (±3) ve 17,06 (±9,8) olarak ölçülmüştür.
Sonuç: Sonuç olarak, Candida türlerinin uygun tedavisi için etkenlerin tür tanımlaması ve izole edilen kökenlerin antifungal duyarlılık testlerinin yapılması ampirik antifungal tedavi politikalarına katkı sağlayacaktır.

Ethical Statement

Çalışmanın etik kurul onayı 2014/119 karar no ile 17.02.2015 tarihli kararında alınmıştır.

Supporting Institution

Düzce Üniversitesi Bilimsel Araştırma Projeleri

Project Number

2015.04.01.353

Thanks

Düzce Üniversitesi Bilimsel Araştırma Projeleri Birimine desteklerinden doleyı teşekkür ediyoruz

References

  • Siddiqui R, Mendiratta DK, Siddiqui AF, Rukadikar A. A study of the association between virulence factors and antifungal susceptibility profile of Candida species recovered from cases of vulvovaginal candidiasis. J Family Med Prim Care. 2023;12(1):152-9.
  • R AN, Rafiq NB. Candidiasis. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2025 [cited 2025 May 02]. Available from: https://www.ncbi.nlm.nih.gov/books/NBK560624/.
  • Achkar JM, Fries BC. Candida infections of the Genitourinary Tract. Clin Microbiol Rev. 2010;23(2):253-73.
  • Lamoth F, Lockhart SR, Berkow EL, Calandra T. Changes in the epidemiological landscape of invasive candidiasis. J Antimicrob Chemother. 2018;73:4-13.
  • El-Kholy MA, Helaly GF, El Ghazzawi EF, El-Sawaf G, Shawky SM. Virulence factors and antifungal susceptibility profile of C. tropicalis isolated from various clinical specimens in Alexandria, Egypt. J Fungi. 2021;7(5):351.
  • Makled AF, Ali SAM, Labeeb AZ, Salman SS, Shebl DZM, Hegazy SG, et al. Characterization of Candida species isolated from clinical specimens: insights into virulence traits, antifungal resistance and molecular profiles. BMC Microbiol. 2024;24(1):388.
  • Clinical and Laboratory Standards Institute (CLSI). Method for antifungal disk diffusion susceptibility testing of yeasts. Approved guideline-second edition, 2nd ed. CLSI document M44-A2. Wayne (PA): CLSI; 2009.
  • Jayachandran AL, Katragadda R, Ravinder T, Vajravelu L, Manorajan L, Hemalatha S, et al. Antifungal susceptibility pattern among Candida species: an evaluation of disc diffusion and micro broth dilution method. J Microbil Infect Dis. 2018;08(03):97-101.
  • Larone DH. Yeasts and yeast like organisms. In: Larone DH. editor. Medically important fungi: a guide to identification. 5th ed. Washington (DC): ASM Press; 2011. p. 117-54.
  • Espinel-Ingroff A, Canton E. Comparison of Neo-Sensitabs tablet diffusion assay with CLSI broth microdilution M38-A and disk diffusion methods for testing susceptibility of filamentous fungi with amphotericin B, caspofungin, itraconazole, posaconazole, and voriconazole. J Clin Microbiol. 2008;46(5):1793-803.
  • Marr KA, Seidel K, White TC, Bowden RA. Candidemia in allogeneic blood and marrow transplant recipients: evolution of risk factors after the adoption of prophylactic fluconazole. J Infect Dis. 2000;181(1):309-16.
  • Ener B. İnvitro antifungal duyarlılık testleri: standardizasyon ve klinik önemi. Mikrobiyol Bül. 1996;30:419-25.
  • Chen X, Shen M, Feng Q, Yang G, Tian R, Yao S, et al. Epidemiological characteristics, antifungal susceptibility, and mortality factors of candidemia in adults at a tertiary teaching hospital in Zunyi, China (2016-2023). BMC Infect Dis. 2025;25(1):726.
  • Zhang W, Song X, Wu H, Zheng R. Epidemiology, risk factors and outcomes of Candida albicans vs. non-albicans candidaemia in adult patients in Northeast China. Epidemiol Infect. 2019;25:147:e277.
  • Bayram Y, Gültepe B, Özlük S, Güdücüoğlu H. Çeşitli klinik örneklerden izole edilen Candida kökenlerinin identifikasyonu ve antifungal duyarlılıklarının araştırılması. Van Tıp Dergisi. 2012;19(4):177-81.
  • Sav H, Demir G, Atalay MA, Koç AN. Klinik örneklerden izole edilen Candida türlerinin değerlendirilmesi. Turk Hij Den Biyol Derg. 2013;70(4):175-80.
  • Pham CD, Iqbal N, Bolden CB, Kuykendall RJ, Harrison LH, Farley MM, et al. Role of FKS Mutations in Candida glabrata: MIC values, echinocandin resistance, and multidrug resistance. Antimicrob Agents Chemother. 2014;58(8):4690-6.
  • Daneshnia F, de Almeida Júnior JN, Ilkit M, Lombardi L, Perry AM, Gao M, et al. Worldwide emergence of fluconazole-resistant Candida parapsilosis: current framework and future research roadmap. Lancet Microbe. 2023;4(6):e470-80.
  • Özkaya E, Çalışkan A, Kirişci Ö, Tüme S. Son üç yılda Kahramanmaraş Necip Fazıl Şehir Hastanesinde kan kültürlerinden izole edilen Candida türlerinin dağılımı ve antifungal duyarlılıkları. Türk Mikrobiyol Cem Derg. 2016;46(2):63-8.
  • Diekema DJ, Messer SA, Hollis RJ, Boyken LB, Tendolkar S, Kroeger J, et al. Evaluation of Etest and disk diffusion methods compared with broth microdilution antifungal susceptibility testing of clinical isolates of Candida spp. against posaconazole. J Clin Microbiol. 2007;45(6):1974-7.
  • Jaruvongvanich V, Worasilchai N, Kongnatthasate K, Jaruvongvanich S, Damkerngsuntorn W, Atikarnbodee D, et al. Correlation between broth microdilution, E-test and disk diffusion methods for testing antifungal susceptibility of Candida species isolated from Thai blood samples. Asian Biomed. 2016;10(1):75-80.
  • Giri S, Kindo AJ. Evaluation of antifungal susceptibility testing in Candida isolates by Candifast and disk-diffusion method. Indian J Pathol Microbiol. 2014;57(4):595-7.
  • Altuncu E, Bilgen H, Çerikcioğlu N, İlki A, Ülger N, Bakır M, et al. Neonatal Candida infeksiyonları ve etkenlerinin antifungal duyarlılıkları. Mikrobiyol Bul. 2010;44:593-603.
  • Yiğit N, Aktaş E. Activities of amphotericin B, fluconazole and voriconazole against Candida bloodstream isolates determined by broth microdilution and disk diffusion methods. Turk Hij Den Biyol Derg. 2014;71(3):131-40.
  • Kubiak DW, Farmakiotis D, Arons V, Hollins RM, Rostas SE, Weiser LM, et al. Utility of in-house fluconazole disk diffusion susceptibility testing in thetreatment of candidemia. Diagn Microbiol Infect Dis. 2016;84(3):223-6.
  • Azevedo AC, Bizerra FC, da Matta DA, de Almeida LP, Rosas R, Colombo AL. In vitro susceptibility of a large collection of Candida strains against fluconazole and voriconazole by using the CLSI disk diffusion assay. Mycopathologia. 2011;171(6):411-6.
  • Pathak AK, Jain NR, Joshi R. Antibiogram of Candida species isolated from mono and multi‑species oral candidal carriage using disk diffusion method. Saudi J Health Sci. 2012;1(3):132-8.
There are 27 citations in total.

Details

Primary Language Turkish
Subjects Medical Microbiology (Other)
Journal Section Research Article
Authors

Hatice Erdem 0009-0007-7813-1595

Emel Akbaş 0000-0001-6589-7734

Nida Akar 0000-0001-6524-8224

Şükrü Öksüz 0000-0002-4893-5564

Project Number 2015.04.01.353
Publication Date September 24, 2025
Submission Date July 21, 2025
Acceptance Date September 1, 2025
Published in Issue Year 2025 Volume: 15 Issue: 3

Cite

Vancouver Erdem H, Akbaş E, Akar N, Öksüz Ş. Çeşitli Klinik Örneklerden İzole Edilen Candida Türlerinin Antifungal Duyarlılıklarının Disk Difüzyon Yöntemiyle Araştırılması. VHS. 2025;15(3):473-8.