Detection of Enzymatic Virulence Factors in Candida albicans and Candida glabrata Isolated from Diabetic Patients

Section: Research Article

Abstract

Candiduria is among the most common infections in diabetic patients, and its incidence has increased markedly in recent decades. Candida spp. are frequently isolated from patients, with non-albicans species being increasingly recognized as important etiological agents. The current study aimed to diagnose Candida spp. isolated from the urine of diabetic women aged 16-72 years and evaluated their ability to produce phospholipase, hemolysin, and esterase enzymes. Statistical analysis of enzymatic activity was performed (P<0.01).  Midstream urine samples were collected from 178 diabetic women, including 86 type 1 and 92 type 2 diabetes. Urine samples were cultured on Sabouraud's Dextrose Agar (SDA) and subsequently identified using conventional methods, including cultural characteristis Gram stainning germ tube formationchlamydospore production, growth at 45℃, and growth on HiCrome Candida Differential Agar M1297A. The present study showed that three different species of Candida were identified: 30 (50%)  C. glabrata isolates, followed by 18 (36%) C. albicans isolates and 2 (4%) C. krusei isolates. Also, the results showed that type 1 diabetic patients are more susceptible to Candida infection. The virulence test revealed that all C. albicans isolates (100%) were positive for phospholipase, hemolysin, and esterase with different production rates. In terms of C. glabrata isolates, one isolate only (3.3%) produced phospholipase, while all isolates (100%) produced hemolysin, and four isolates (13.3%) produced esterase.

References

  1. M. Paknejadi, M. Bayat and V. Razavilar, “Investigating the Frequency of Candida glabrata in Diabetic Women of Tehran with Recurrent and Non-recurrent Vulvovaginal Candidiasis Using PCR-RFLP Assay”. J. Med. Microbiol. Infect. Dis., vol. 7, no. 1, pp. 44-51, 2019, doi: 10.29252/JoMMID.7.1.2.44.
  2. G. R. Thompson III, S. C. Chen, W. A. Alfouzan, K. Izumikawa, A. L., Colombo and J. Maertens, “A global perspective of the changing epidemiology of invasive fungal disease and real-world experience with the use of isavuconazole”. Med. Mycol., vol. 62, no. 9, p. myae083, 2024, https://doi.org/10.1093/mmy/myae083.
  3. M. Gajdács, I. Dóczi, M. Ábrok, A. Lázár, and K. Burián, “Epidemiology of candiduria and candida urinary tract infections in inpatients and outpatients: results from a 10- year retrospective survey”. Cent European J Urol., vol. 72, no. 2, pp. 209- 214, 2019, DOI: 10.5173/ceju.2019.1909.
  4. V. C. Abishek, K. Mohanram, and R. Preethy, “Prevalence of candiduria and characterisation of Candida species in tertiary care center”. Saudi J. Pathol. Microbiol., vol. 4, no. 7, pp. 573- 576, 2019, https://doi.org/10.21276/SJPM.2019.4.7.18.
  5. Z. He, C. Su, Y. Bi, Y. Cheng, D. Lei, and F. Wang, “Evaluation of a novel Laboratory Candiduria Screening protocol in the intensive care unite”. Infect Drug Resist., vol. 14, pp. 489- 496, 2021, DOI: 10.2147/IDR.S289885.
  6. A. Gharanfoli, E. Mahmoudi, R. Torabizadeh, F. Katiraii, and S. Faraji, “Isolation, characterization, and molecular identification of Candida species from urinary tract infections”. Curr. Med. Mycol., vol. 5, no. 2, pp. 33-36, 2019, doi: 10.18502/cmm.5.2.1159.
  7. J. Pemán, and A. Ruiz-Gaitán, “Candidemia from urinary tract source: the challenge of candiduria”. Taylor and Francis, vol. 46, no. 5, pp. 243- 245, 2018, https://doi.org/10.1080/21548331.2018.1538623.
  8. M. Moazeni, and M. Nabili, “Identification of Candida Species Isolated from Hospitalized Patients with Candiduria”. Med. Lab. J., vol. 16, no. 2, pp. 13-20, 2022, doi: 10.61186/mlj.16.2.13.
  9. S. Manzoor, M. Aziz, and A. S. Sheikh, “Identification and Characterization of Candida on CHROMagar in pregnant women of Multan, Pakistan”. J. Womens Health Care, vol. 7, no. 2, p. 1000424, 2018, DOI: 10.4172/2167-0420.1000424 .
  10. G. Aboagye, S. Waikhom, E. A. Asiamah, C. O. Tettey, H. Mbroh, C. Smith, G.Y. Osei, K. Asafo Adjei, and R. H. Asmah, “Antifungal susceptibility profiles of Candida and non-albicans species isolated from pregnant women: implications for emerging antimicrobial resistance in maternal health”.
  11. Microbiol. Spectr., vol. 13, no. 7, pp.e00787-25. https://doi.org/10.1128/spectrum.00787-25.
  12. A. R. Hameed, S. M. Ali, and T. L. Ahmed, “Biological study of Candida species and virulence factor”. Int. J. Adv. Res. Eng. Technol., vol. 1, no. 4,pp. 8-17, 2018.
  13. N. N. Jammil, and M. M. Yehia, “Virulence factors of yeasts isolated from oral lesions”. Ann. Coll. Med. Mosul, vol. 41, no. 2, pp. 170- 177, 2019, https://doi.org/10.33899/mmed.2020.164160.
  14. D.L. Zuza-Alves, S. de Medeiros, L. de Souza, W. Silva- Rocha, E. Francisco, M. De Araujo, R. Lima- Neto, R. Neves, A. Melo, G. Chaves, “Evaluation of virulence factors in vitro, resistance to osmotic stress and antifungal susceptibility of Candida tropicalis isolated from the coastal environment of northeast Brazil”. Front. Microbiol., vol. 7, p. 1783, 2016, DOI: 10.3389/fmicb.2016.01783.
  15. M. Maheronnaghsh, M. Fatahinia, P. Dehghan, A. Mahmoudabadi, and M. Kheirkhah, “Comparison of virulence factors of different Candida specis isolated from the oral cavity of cancer patients and normal individuals”. Jundishapur J Microbiol., vol. 12, no. 5, p. :e91556, 2019, https://doi.org/10.5812/jjm.91556.
  16. C. P. Kurtzman, and J. W. Fell, The Yeasts, a Taxonomic Study. 4th ed., Elsevier Science B. V., The Netherland; 1998.
  17. B. Forbes, D. Sahm, and A. Weissfeld, Bailey and Scott's Diagnostic Microbiology. 12th ed., Mosby. Inc., Elsevier; 2007, p. 710.
  18. E. Pinjon, D. Sullivan, I. Salkin, D. Shanley, and D. Coleman, “Simple, inexpensive, reliable method for differentiation of Candida dubliniensis from Candida albicans”. J. Clin. Microbiol., vol. 36, no. 7, pp. 2093- 2095, 1998, DOI: 10.1128/JCM.36.7.2093-2095.1998.
  19. M. Price, I. Wilkinson, and L. Gentry, “Plate method for detection of phospholipase activity in Candida albcans”. Sabouraudia, vol. 20, pp. 7- 14, 1982, DOI: 10.1080/00362178285380031.
  20. C. Kumar, S. Kumar, and T. Menon, “Phospholipase and Proteinase activities of clinical isolates of Candida from immuhocompromised patients”. Mycopathologia, vol. 161, no. 4, 213- 218, 2006, doi: 10.1007/s11046-005-0157-4.
  21. S. Ying, S. Qing, and L. Chunyang, “The effect of gentian violet on virulent properties of Candida albicans”. Mycopathologia, vol. 169, no. 4, pp. 279- 285, 2010, doi: 10.1007/s11046-009-9258-9.
  22. M. Noori, M. Dakhili, A. Sepahvand, and N. Davari, “Evaluation of esterase and hemolysin activities of different Candida species isolated from vulvovaginitis cases in Lorestan Province, Iran”. Curr. Med. Mycol.,vol. 3, no.4, pp. 1-5, 2017, doi: 10.29252/cmm.3.4.1 .
  23. G. Luo, L. P. Samaranayake, J.Yau, “ Candida species exhibit differential in vitro hemolytic activites”. J. Clin. Microbiol., vol. 39, no. 8, 2001, https://doi.org/10.1128/jcm.39.8.2971-2974.2001.
  24. J. M. Manns, D. M. Mosser, and H.R. Buckley, “Production of hemolytic factor by Candida albicans”. Infect. Immun., vol. 62, no. 11, pp. 5154- 5156, 1994, https://doi.org/10.1128/iai.62.11.5154-5156.1994.
  25. S.C. Deorukhkar, S. Saini, N.A. Raytekar, and M.D. Sebastian, “Catheter associated urinary tract Candida infections in intensive care unite patients”. J. Clin. Microbiol. Biochem. Technol.,vol. 2, no.1, pp. 015-017, 2016, doi: 10.17352/jcmbt.000010.
  26. M. Falahati, S. Farahyar, L. Akhlaghi, S. Mahmoudi, K. Sabzian, M. Yarahmadi, and R. Aslani, “Characterization and identification of candiduria due to candida species in diabetic patients”. Curr. Med. Mycol., vol. 2, no. 3, pp. 10- 14, 2016, doi: 10.18869/acadpub.cmm.2.3.10.
  27. R. Altınbaş, and Y. Bildirici, “Distribution of yeasts in fungal urinary tract infections from a tertiary care hospital”. J. Med. Palliat. Care, vol. 4, no. 4, pp. 296-301, 2023, DOI: 10.47582/jompac.1325399.
  28. C. F. Rodrigues, M. E. Rodrigues, and M. Henriques, “Candida sp. Infections in patients with diabetes mellitus”. J. Clin. Med., vol. 8, no. 76, pp. 1-41, 2019, doi:10.3390/jcm8010076 .
  29. B. Saud, P. Bajgain, G. Paudel, V. Shrestha, D. Bajracharya, S. Adhikari, G. Dhungana, and M. Awasthi, “Fungal infection among diabetic and Nondiabetic individuals in Nepal.” Interdiscip. Perspect. Infect. Dis., vol. 2020, no. 1,pp. 7949868, 2020, https://doi.org/10.1155/2020/7949868.
  30. G. Yismaw, D. Asrat, Y. Woldeamanuel, and C. Unakal, “Prevalence of candiduria in diabetic patients attending Gondar university hospital, Gondar, Ethiopia.” Iran J. Kidney Dis., vol. 7, no. 2, pp. 102- 107, 2013.
  31. A. Esmailzadeh, H. Zarrinfar, A.M. Fata, and T. Sen, “High prevalence of candiduria due to non- albicans Candida species among diabetic patients: A matter of concern?” J. Clin. Lab. Anal.,vol. 32, no. 6, pp. e 22343, 2017, doi: 10.1002/jcla.22343.
  32. H. Nouraei, M.G. Jahromi, L.R. Jahromi, K. Zomorodian, and K. Pakshir, “Potential pathogenicity of Candida species isolated from oral cavity of patients with diabetes mellitus”. Biomed. Res. Int., vol. 2021, no. 1, pp. 9982744, 2021, https://doi.org/10.1155/2021/9982744.
  33. S.P. Tsang, F. Chu, W.K. Leung, L.J. Jin, L.P. Samaranayake, and S.C. Siu, “Phospholipase, Proteinase and haemolytic activities of Candida albicans isolated from oral cavities of patients with type 2 diabetes mellitus”. J. Med. Microbiol., vol. 56, no. 10, pp. 1393-1398, 2007, https://doi.org/10.1099/jmm.0.47303-0.
  34. A. Mattei, S. Alves, C. Severo, L. Guazzelli, F. Oliveira, and L. Severo, “Determination of germ tube, phospholipase, and proteinase production by bloodstream isolates of Candida albicans”. Rev. Soc. Bras. Med. Trop., vol. 46, no. 3, pp. 340- 342, 2013, https://doi.org/10.1590/0037-8682-0045-2013 .
  35. Y. Hassan, S.Y. Chew, L. Than, “ Candida glabrata: Pathogenicity and resistance mechanisms for adaptation and surviral”. J. Fungi,vol. 7, no. 8, p. 667, 2021, https://doi.org/10.3390/jof7080667.
  36. H.K. Malcok, E. Aktas, A. Ayyildiz, N. Yigit, H. Yazgi, “ Hemolytic activities of the Candida species in liquid medium”. Eurasian J. Med., vol. 41, no. 2, pp. 95–98, 2009.
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“Detection of Enzymatic Virulence Factors in Candida albicans and Candida glabrata Isolated from Diabetic Patients”, JES, vol. 35, no. 2, pp. 126–135, Apr. 2026, doi: 10.33899/jes.v35i2.60528.
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[1]
“Detection of Enzymatic Virulence Factors in Candida albicans and Candida glabrata Isolated from Diabetic Patients”, JES, vol. 35, no. 2, pp. 126–135, Apr. 2026, doi: 10.33899/jes.v35i2.60528.