Detection of some virulence factors among Candida albicans isolated from patients and prevalence of candidalysin gene ECE1
Main Article Content
Abstract
Candida albicans is a common cause of respiratory infection and oral candidiasis in people; it is an opportunistic yeast pathogen and a major cause of morbidity and mortality in the immunocompromised persons and causes superficial infections of mucosal surfaces which affect millions of people throughout world. The main goal of this study was investigating the prevalence of some virulence factors which has the ability to configure biofilm formation, proteinase, hemolysin production among C.albicans isolates that include prevalence of candidalysin gene Eec1. Samples were collected during the period May and August of 2022 from 280 samples (swabs) of different ages and sexes of non-duplicated Iraqi patients suffering from oral candidiasis and respiratory diseases. The results showed that 102 were positive samples, 58(56.86%) from oral cavity and 44 (43.14%) from respiratory tract, while 178 of them were negative. Candida isolates were identified using conventional methods by grown on HiCrome Candida medium, germ tube production, chlamydospore formation and confirmed using VITEK-2 system, susceptibility of Candida isolates to antifungal drugs was examined by disk diffusion method, performed as recommended by (CLSI) M44-A document. The isolates showed a high level of susceptibility to Amphotericin-B (93.20%), Nystatin (90.20%), and Clotrimazole (85.92%). Prevalence of Candidalysin gene Eec1 among 70 isolates of C. albicans was investigated using polymerase chain reaction (PCR) technique, the results revealed that 41(58.57%) were harboring Ece1 gene for the oral cavity and respiratory tract. Only 34 (48.57%) C. albicans isolates were strong producer of biofilm, while 30 (42.86%) isolates produced proteinase, 20 (28.57%) of isolates had the ability to hemolyze the blood.
Received 10/03/2023
Revised 07/04/2023
Accepted 09/04/2023
Published Online First 20/09/2023
Article Details
This work is licensed under a Creative Commons Attribution 4.0 International License.
How to Cite
References
Mora C, Tittensor DP, Adl S, Simpson AG, Worm B. How many species are there on Earth and in the ocean?. PLoS Biol. 2011; 9 (8): e1001127. https://doi.org/10.1371/journal.pbio.1001127.
Gnat S, Łagowski D, Nowakiewicz A, Dyląg M. A global view on fungal infections in humans and animals: opportunistic infections and microsporidioses. J Appl Microbiol. 2021 1; 131(5): 2095-113. https://doi.org/10.1111/jam.15032
Naglik JR, Gaffen SL, Hube B. Candidalysin: discovery and function in Candida albicans infections. Curr Opin Microbiol. 2019; 52: 100-109. https://doi.org/10.1016/j.mib.2019.06.002
Hibbett DS, Blackwell M, James TY, Spatafora JW, Taylor JW, Vilgalys R. Phylogenetic taxon definitions for Fungi, Dikarya, Ascomycota and Basidiomycota. IMA Fungus. 2018; 9: 291-298 https://doi.org/10.5598/imafungus.2018.09.02.05.
Russell CM, Schaefer KG, Dixson A, Gray ALH, Pyron RJ, Alves DS, et al. The Candida albicans virulence factor candidalysin polymerizes in solution to form membrane pores and damage epithelial cells. Elife. 2022; 29: 11: e75490. https://doi.org/10.7554/eLife.75490
Richardson JP, Brown R, Kichik N, Lee S, Priest E, Mogavero S, et al. Candidalysins Are a New Family of Cytolytic Fungal Peptide Toxins. ASM JmBio. 2022 Feb 22; 13(1): e0351021. https://doi.org/10.1128/mbio.03510-21
Chen H, Zhou X, Ren B, Cheng L. The regulation of hyphae growth in Candida albicans. Virulence. 2020; 11(1): 337-348. https://doi.org/10.1080/21505594.2020.1748930
Mogavero S, Sauer FM, Brunke S, Allert S, Schulz D, Wisgottet S, et al. Candidalysin delivery to the invasion pocket is critical for host epithelial damage induced by Candida albicans. Cell Microbiol. 2021; 23(10): e13378. https://doi.org/10.1111/cmi.13378
Vaňková E, Kašparová P, Dulíčková N, Čeřovský V. Combined effect of lasioglossin LL-III derivative with azoles against Candida albicans virulence factors: biofilm formation, phospholipases, proteases and hemolytic activity. FEMS Yeast Res. 2020 1; 20(3): foaa020. https://doi.org/10.1093/femsyr/foaa020
Aldossary MA, Almansour NA, Abdulraheem BS. Isolation and identification of Candida species from the oral cavity of cancer patients undergoing chemotherapy in Basrah, Iraq. Biol Agr Healthc. 2016; 6(18): 22-30.
Elmer W K, Stephen D A, William M J. Laboratory approach to the diagnosis of fungal infections.1992. 14th edition, J P Lipincott Company, Philadelphia. pp. 387-840.
Wayne PA. Clinical and Laboratory Standards Institute: Performance standards for antimicrobial susceptibility testing: 27th ed informational supplement. CLSI document M100-S20. PA 2017.
Deravi N, Fathi M, Tabatabaeifar SN, Ahmadi B, Shokoohi G, Ansari S. Azole antifungal resistance in Candida albicans and Candida glabrata isolated from vulvovaginal candidiasis patients. Arch Clin Infect Dis. 2021; 16(2). https://dx.doi.org/10.5812/archcid.106360
Shoukat M, Ullah F, Tariq MN, Din G, Khadija B, Faryal R. Profiling of potential pathogenic Candida species in obesity. Microb Pathog. 2023; 174: 105894. https://dx.doi.org/10.1016/j.micpath.2022.105894
Staib F. Serum-proteins as nitrogen source for yeastlike fungi. Sabouraudia. 1965; 4(3): 187-93. https://doi.org/10.1080/00362176685190421
Swidergall M, Solis NV, Millet N, Huang MY, Lin J, Lin J, et al. Activation of EphA2-EGFR signaling in oral epithelial cells by Candida albicans virulence factors. PLoS Pathog. 2021; 20: 17(1): e1009221. https://doi.org/10.1371/journal.ppat.1009221
Jin Y, Yip HK, Samaranayake YH, Yau JY, Samaranayake LP. Biofilm-forming ability of Candida albicans is unlikely to contribute to high levels of oral yeast carriage in cases of human immunodeficiency virus infection. J Clin Microbiol. 2003; 41(7): 2961-7. https://doi.org/10.1128/jcm.41.7.2961-2967.2003
Davarzani F, Saidi N, Besharati S, Saderi H, Rasooli I, Owlia P. Evaluation of Antibiotic Resistance Pattern, Alginate and Biofilm Production in Clinical Isolates of Pseudomonas aeruginosa. Iran J Public Health. 2021; 50(2): 341-349. https://doi.org/10.18502/ijph.v50i2.5349
Daniel W W, Cross C L. Biostatistics: A Foundation for Analysis in the Health Sciences. 11th Edition, John Wiley & Sons, Hoboken. 2018. 720 pp.
Sardi JCO, Scorzoni L, Bernardi T, Fusco-Almeida AM, Mendes Giannini MJS. Candida species: current epidemiology, pathogenicity, biofilm formation, natural antifungal products and new therapeutic options. J Med Microbiol. 2013; 62(Pt1): 10-24. https://doi.org/10.1099/jmm.0.045054-0
Pinto-Almazán R, Frías-De-León MG, Fuentes-Venado CE, Arenas R, González-Gutiérrez L, Chávez-Gutiérrez E, et al. Frequency of Candida spp. in the Oral Cavity of Asymptomatic Preschool Mexican Children and Its Association with Nutritional Status. Children (Basel). 2022; 9(10): 1510. https://doi.org/10.3390/children9101510
Taverne-Ghadwal L, Kuhns M, Buhl T, Schulze MH, Mbaitolum WJ, Kersch L, et al. Epidemiology and prevalence of Oral candidiasis in HIV patients from Chad in the post-HAART era. Front Microbiol. 2022; 13. https://doi.org/10.3389/fmicb.2022.844069
Vila T, Sultan AS, Montelongo-Jauregui D, Jabra-Rizk MA. Oral candidiasis: a disease of opportunity. J Fungi. 2020; 6(1): 15. https://doi.org/10.3390/jof6010015
Patil S, Rao RS, Majumdar B, Anil S. Clinical Appearance of Oral Candida Infection and Therapeutic Strategies. Front Microbiol. 2015; 6: 1391. https://doi.org/10.3389/fmicb.2015.01391
Habib KA, Najee EN, Abood MS. Identification of Candida species Isolated From Vulvovaginal Candidiasis Patients by Chromgen agar and PCR-RFLP Method. Baghdad Sci J. 2016; 13(2): 291-297. https://doi.org/10.21123/bsj.2016.13.2.0291
Mulet Bayona JV, Salvador García C, Tormo Palop N, Valentín Martín A, González Padrón C, Colomina Rodríguez J, et al. Novel Chromogenic Medium CHROMagarTM Candida Plus for Detection of Candida auris and Other Candida Species from Surveillance and Environmental Samples: A Multicenter Study. J Fungi (Basel). 2022; 8(3): 281. https://doi.org/10.3390/jof8030281
Patel M. Oral Cavity and Candida albicans: Colonisation to the Development of Infection. Pathogens. 2022; 11(3): 335. https://doi.org/10.3390/pathogens11030335
Quinn PJ, Markey BK, Leonard FC, Hartigan P, Fanning S, Fitzpatrick E. Veterinary microbiology and microbial disease. 2nd edition. Wiley-Blackwell, 2011, Chichester, West Sussex, UK.
Chaffin WL, López-Ribot JL, Casanova M, Gozalbo D, Martínez JP. Cell wall and Secreted proteins of Candida albicans: identification, function, and expression. Microbiol Mol Biol Rev. 1998; 62(1):130-80. https://doi.org/10.1128/MMBR.62.1.130-180.1998
Wang X, Mohammad IS, Fan L, Zhao Z, Nurunnabi M, Sallam MA, et al. Delivery strategies of amphotericin B for invasive fungal infections. Acta Pharmaceutica Sinica B. 2021; 11(8): 2585-604. https://doi.org/10.1016/j.apsb.2021.04.010
Mohamed BJ and AL. Thwani A. The susceptibility test of vaginal yeasts and their relationship with the age in Iraqi women. Baghdad Sci J. 2010; 7(3): 1174-1180. https://doi.org/10.21123/bsj.2010.7.3.1174-1180
Bilal H, Hou B, Shafiq M, Chen X, Shahid MA, Zeng Y. Antifungal susceptibility pattern of Candida isolated from cutaneous candidiasis patients in eastern Guangdong region: A retrospective study of the past 10 years. Front Microbiol. 2022; 13: 981181. https://doi.org/10.3389/fmicb.2022.981181
Wang E, Farmakiotis D, Yang D, McCue DA, Kantarjian HM, Kontoyiannis DP, et al. The ever-evolving landscape of candidaemia in patients with acute leukaemia: non-susceptibility to caspofungin and multidrug resistance are associated with increased mortality. J Antimicrob Chemother. 2015; 70(8): 2362-8. https://doi.org/10.1093/jac/dkv087
Al-Dahlaki NOM, and Al-Qaysi SAS. Production and Identification of Secondary Metabolite Gliotoxin-Like Substance Using Clinical Isolates of Candida spp. Microbiol Biotechnol Lett. 2022; 50(4): 488-500. https://doi.org/10.48022/mbl.2209.09007
Magaldi S, Mata-Essayag S, Hartung de Capriles C, Perez C, Colella MT, Olaizola C. Well diffusion for antifungal susceptibility testing. Int J Infect Dis. 2004; 8(1): 39-45. https://doi.org/10.1016/j.ijid.2003.03.002
Yasu T, Konuma T, Kuroda S, Takahashi S, Tojo A. Effect of Cumulative Intravenous Voriconazole Dose on Renal Function in Hematological Patients. Antimicrob Agents Ch. 2018; 62(9): e00507-18. https://doi.org/10.1128/aac.00507-18
Jenks JD, Salzer HJ, Prattes J, Krause R, Buchheidt D, Hoenigl M. Spotlight on isavuconazole in the treatment of invasive aspergillosis and mucormycosis: design, development, and place in therapy. Drug Des Devel Ther. 2018; 12: 1033-1044. https://doi.org/10.2147/dddt.s145545
Othman KI, Abdullah SM, Majid AM. Isolation and Identification Candida spp. from Urine and Antifungal Susceptibility Test. Iraqi J Sci. 2018; 59(4B): 1981-1988. http://doi.org/10.24996/ijs.2018.59.4B.3
Vadiraj S, Nayak R, Choudhary GK, Kudyar N, Spoorthi BR. Periodontal pathogens and respiratory diseases- evaluating their potential association: a clinical and microbiological study. J Contemp Dent Pract. 2013; 14(4): 610-5. https://doi.org/10.5005/jp-journals-10024-1373
Di Cosola M, Cazzolla AP, Charitos IA, Ballini A, Inchingolo F, Santacroce L. Candida albicans and oral carcinogenesis. A brief review. J Fungi. 2021; 7(6): 476. https://doi.org/10.3390/jof7060476
Cangui-Panchi SP, Ñacato-Toapanta AL, Enríquez-Martínez LJ, Reyes J, Garzon-Chavez D, Machado A. Biofilm-forming microorganisms causing hospital-acquired infections from intravenous catheter: A systematic review. Curr Res Microb Sci. 2022; 3: 100175. https://doi.org/10.1016/j.crmicr.2022.100175
Bekkal Brikci Benhabib O, Boucherit-Otmani Z, Boucherit K, Djediat C. Interaction in a dual-species biofilm of Candida albicans and Candida glabrata co-isolated from intravascular catheter. Microb Pathog. 2021; 152: 104613. https://doi.org/10.1016/j.micpath.2020.104613
Lohse MB, Gulati M, Johnson AD, Nobile CJ. Development and regulation of single- and multi-species Candida albicans biofilms. Nat Rev Microbiol. 2018; 16(1): 19-31. https://doi.org/10.1038/nrmicro.2017.107
Rodríguez-Cerdeira C, Martínez-Herrera E, Carnero-Gregorio M, López-Barcenas A, Fabbrocini G, Fida M, El-Samahy M, et al. Pathogenesis and Clinical Relevance of Candida Biofilms in Vulvovaginal Candidiasis. Front Microbiol. 2020; 11: 544480. https://doi.org/10.3389/fmicb.2020.544480
Mohammed NA, Ajah HA, Abdulbaqi NJ. Detection The Prevalence of Adhesins and Extracellular hydrolytic enzymes genes in Candida albicans Biofilm Formation. Iraq J Sci. 2017; 58(2C): 988-1000. https://doi.org/10.24996.ijs.2017.58.2C.3
Atriwal T, Azeem K, Husain FM, Hussain A, Khan MN, Alajmi MF, Abid M. Mechanistic understanding of Candida albicans biofilm formation and approaches for its inhibition. Front Microbiol. 2021; 12: 638609. https://doi.org/10.3389/fmicb.2021.638609
Mello VG, Escudeiro H, Weckwerth AC, Andrade MI, Fusaro AE, de Moraes EB, Ruiz LD, Baptista IM. Virulence factors and antifungal susceptibility in Candida species isolated from dermatomycosis patients. Mycopathologia. 2021; 186(1): 71-80. https://doi.org/10.1007/s11046-020-00509-x
Talapko J, Juzbašić M, Matijević T, Pustijanac E, Bekić S, Kotris I, et al. Candida albicans- the virulence factors and clinical manifestations of infection. J Fungi. 2021; 7(2): 79. https://doi.org/10.3390/jof7020079
Yang HP, Tsang PC, Pow EH, Lam OL, Tsang PW. Candida albicans Secreted Aspartic Protease 7 is Essential for Damage of Human Oral Epithelial Cells. J Bacteriol Mycol. 2021; 8(5): 1182.
Staniszewska M. Virulence factors in Candida species. Curr Protein Pept Sci. 2020; 21(3): 313-23. https://doi.org/10.2174/1389203720666190722152415
Vieira E, Cunha SC, Ferreira IM. Characterization of a potential bioactive food ingredient from inner cellular content of brewer’s spent yeast. Waste Biomass Valori. 2019; 10: 3235-3242. https://doi.org/10.1007/s12649-018-0368-9
Ramos-Pardo A, Castro-Álvarez R, Quindós G, Eraso E, Sevillano E, Kaberdin VR. Assessing pH-dependent activities of virulence factors secreted by Candida albicans. Microbiologyopen. 2023; 12(1): e1342. https://doi.org/10.1002/mbo3.1342
Abeed FK, Alrubayae IM. Evaluation of virulence factors of clinical yeast isolates from nosocomial fungal infections with the determination of their antifungal susceptibility profile. Iran J Ichthyol. 2022; 15(9): 61-8.
Nouraei H, Pakshir K, ZareShahrabadi Z, Zomorodian K. High detection of virulence factors by Candida species isolated from bloodstream of patients with candidemia. Microb Pathog. 2020; 149: 104574. https://doi.org/10.1016/j.micpath.2020.104574
Linares CE, de Loreto ES, Silveira CP, Pozzatti P, Scheid LA, Santurio JM, et al. Enzymatic and hemolytic activities of Candida dubliniensis strains. Rev Inst Med Trop Sao Paulo. 2007; 49(4): 203-6. https://doi.org/10.1590/s0036-46652007000400001
Nouraei H, Pakshir K, ZareShahrabadi Z, Zomorodian K. High detection of virulence factors by Candida species isolated from bloodstream of patients with candidemia. Microb Pathog. 2020; 149: 104574. https://doi.org/10.1016/j.micpath.2020.104574
Salvin SB. Hemolysin from the yeast-like phases of some pathogenic fungi. Proc Soc Exp Biol Med. 1951; 76(4): 852-4. https://doi.org/10.3181/00379727-76-18653
Lee S, Kichik Rodriguez N, Hepworth O, Richardson J, Naglik J. In Vitro Biophysical Characterization of Candidalysin: A Fungal Peptide Toxin. In In Vitro Biophysical Characterization of Candidalysin: A Fungal Peptide Toxin. SpringerLink. 2022. p. 163-176. https://doi.org/10.1007/978-1-0716-2549-1_12
Manns JM, Mosser DM, Buckley HR. Production of a hemolytic factor by Candida albicans. Infect Immun. 1994; 62(11): 5154-6. https://doi.org/10.1128/iai.62.11.5154-5156.1994
Richardson JP, Brown R, Kichik N, Lee S, Priest E, Mogavero S, et al. Candidalysins Is a New Family of Cytolytic Fungal Peptide Toxins. ASM J mBio. 2022; 13(1): e0351021. https://doi.org/10.1128/mbio.03510-21
Mogavero S, Sauer FM, Brunke S, Allert S, Schulz D, Wisgott S, et al. Candidalysin delivery to the invasion pocket is critical for host epithelial damage induced by Candida albicans. Cell Microbiol. 2021; 23(10): e13378. https://doi.org/10.1111/cmi.13378
Roy U, Kornitzer D. Heme-iron acquisition in fungi. Curr Opin Microbiol. 2019 Dec 1; 52: 77-83. https://doi.org/10.1016/j.mib.2019.05.006
Richardson JP, Mogavero S, Moyes DL, Blagojevic M, Krüger T, Verma AH, et al. Processing of Candida albicans Ece1p Is Critical for Candidalysin Maturation and Fungal Virulence. ASM J mBio. 2018; 9(1): e02178-17. https://doi.org/10.1128/mBio.02178-17
Richardson JP, Willems HME, Moyes DL, Shoaie S, Barker KS, Tan SL, et al. Candidalysin Drives Epithelial Signaling, Neutrophil Recruitment, and Immunopathology at the Vaginal Mucosa. Infect Immun. 2018; 86(2): e00645-17. https://doi.org/10.1128/IAI.00645-17
Macias-Paz IU, Pérez-Hernández S, Tavera-Tapia A, Luna-Arias JP, Guerra-Cárdenas JE, Reyna-Beltrán E. Candida albicans the main opportunistic pathogenic fungus in humans. Rev Argent Microbiol. 2022; https://doi.org/10.1016/j.ram.2022.08.003
Bain JM, Louw J, Lewis LE, Okai B, Walls CA, Ballou ER, et al. Candida albicans hypha formation and mannan masking of β-glucan inhibit macrophage phagosome maturation. ASM J mBio. 2014; 5(6):e01874. https://doi.org/10.1128/mBio.01874-14
Willems HME, Lowes DJ, Barker KS, Palmer GE, Peters BM. Comparative Analysis of the Capacity of the Candida Species To Elicit Vaginal Immunopathology. Infect Immun. 2018; 86(12): e00527-18. https://doi.org/10.1128/IAI.00527-18