Document Type : Original Article
1 Department of biology /College of Education for Pure Science / Ibn Al-Haitham, Baghdad, Iraq
2 Department of Biotechnology, College of Biotechnology, Al-Nahrain University, Baghdad, Iraq
Background: Candida Berkh. (1923) occurs naturally in the body. But it becomes opportunistic fungi, meaning that it infects humans when there is any weakening of the immune system, such as exposure to chemotherapy, diabetes, or organ transplantation. Most species of Candida grow at a temperature between 20-40 °C and have a pH of 3-8. Human pathogens of Candida species include C. albicans, C. glabrata, C. lusitaniae, C. parapsilosis, C. tropicalis and C. utili. C. albicans has many virulence factors that facilitate injury process. Virulence factors are considered as a measure of pathogenicity, and it is in the form of fungal toxins, enzymes, or cell structures that facilitate infection, as well as pathogen resistance in different conditions. This study aimed to investigate the frequency of some secreted aspartyl proteinases (SAP) genes from some Candida species isolated from diabetic patients
Methods: Candida spp. were identified on CHROMagar medium by color of each species after incubation at 37 °C for 48 hours such as: C. krusei, C. albicans, C. glabrata, and C. tropicalis. Germ tube formation test was used to distinguish between germ tube-forming species, such as C. albicans and C. dubliniensis , from those that do not produce germ tube. A portion of the isolation was taken and placed in a test tube containing 0.5 ml of serum, and then incubated at 37 °C for 2-4 hours. A drop of serum was taken out and examined under a microscope for the presence of the germ tube. Growth at 45 °C, has the advantage of distinguishing between C. albicans and C. dubliniensis by culturing Candida spp. on SDA medium and, incubation for 48-72 hours at 45 °C.
Results: The results of isolation and diagnosis showed that Candida species grew at 37 °C for 48 hours on SDA medium, the shape of the colonies was round or oval-convex, and creamy white, shiny and smooth color. The results of the microscopic examination of the cells were spherical or oval, C. albicans was distinguished by its ability to grow at 45 °C and its ability to form the germ tube. The surface growth test showed the ability of C. tropicalis to grow surface on SDB medium. The medium of CHROMagar showed that C. albicans were light green in color. C. glabrata showed a pale pink color, while C. tropecalis showed a blue color and C. dublineiensis was dark green. DNA extracted samples, including 20 isolates, were used for four species of Candida, 1-9 C. albicans, 10-16 C. glabrata, 17-18 C. dubliniensis and 19-20 C. tropicalis. Parts of the body (mouth, vagina, Urine) detect SAP1 and SAP7 genes by PCR, and after electrophoresis of the PCR product, the results were positive for all Candida species studied as all isolates that contained SAP1 (578bp) and SAP7 (466bp).
Conclusion: The SAP genes are among the main virulence factors for the occurrence of candidiasis. SAP1 and SAP7 genes are responsible for digesting and breaking down protein to penetrate the host's tissues.
- Organization W H. (2016). World health statistics 2016: monitoring health for the SDGs sustainable development goals: World Health Organization. 161 pages,
- Okur M E, Karantas I D, Siafaka P I. (2017). Diabetes Mellitus: a review on pathophysiology, current status of oral pathophysiology, current status of oral medications and future perspectives. ACTA Pharmaceutica Sciencia, 55(1): 61-82. https://doi.org/10.23893/1307-2080.APS.0555
- khajeh h, Bahari A, Rashki A. (2021). TCF7L2 Polymorphisms in Type 2 Diabetes, Insight from HRM and ARMS Techniques. Int. J. Adv. Biol. Biomed. Res., 9(3): 204-214. https://doi.org/10.22034/ijabbr.2021.525681.1351
- Akortha E, Nwaugo V, Chikwe N. (2009). Antifungal resistance among Candida species from patients with genitourinary tract infection isolated in Benin City, Edo state, Nigeria. African Journal of Microbiology Research, 3(11): 694-699.
- Al-Oebady M A H. (2015). Isolation and identification of Candida species from vaginal, urine and oral swabs by chromagar Candida. International Journal of Advanced Research, 3(1): 948-956.
- Muhsen T A, Hawar S N, Mahdi T S, Khaleel R. (2020). Effect of eucalyptus and myrtus extracts identification by gas chromatography-mass spectrometry on some species of candida as a model of medical plants. Ann. Trop. Med. and Public Health, 23(S10): 1-11. http://doi.org/10.36295/ASRO.2020.231032
- Silva S, Negri M, Henriques M, Oliveira R, Williams D W, Azeredo J. (2011). Adherence and biofilm formation of non-Candida albicans Candida species. Trends in microbiology, 19(5): 241-247. https://doi.org/10.1016/j.tim.2011.02.003
- Fazeli-Nasab B, Rahmani A F, Valizadeh M, Khajeh H, Beigomi M. (2021). Evaluation of Antimicrobial Activity of Some Medicinal Plants on Human Standard Bacteria and Candida albicans. [Research Article]. Gene Cell Tissue, 8(3): e113092. https://doi.org/10.5812/gct.113092
- Fazeli-Nasab B, Sayyed R Z, Sobhanizadeh A. (2021). In Silico Molecular Docking Analysis of α-Pinene: An Antioxidant and Anticancer Drug Obtained from Myrtus communis. Int. J. Cancer Manag., 14(2): e89116. https://doi.org/10.5812/ijcm.89116
- Sariguzel F M, Berk E, Koc A N, Sav H, Demir G. (2015). Investigation of the relationship between virulence factors and genotype of Candida spp. isolated from blood cultures. The Journal of Infection in Developing Countries, 9(08): 857-864. https://doi.org/10.3855/jidc.5359
- Pandey N, Gupta M K, Tilak R. (2018). Extracellular hydrolytic enzyme activities of the different Candida spp. isolated from the blood of the Intensive Care Unit-admitted patients. Journal of laboratory physicians, 10(04): 392-396. https://doi.org/10.4103/JLP.JLP_81_18
- Naglik J R, Rodgers C A, Shirlaw P J, Dobbie J L, Fernandes-Naglik L L, Greenspan D, Agabian N, Challacombe S J. (2003). Differential expression of Candida albicans secreted aspartyl proteinase and phospholipase B genes in humans correlates with active oral and vaginal infections. The Journal of infectious diseases, 188(3): 469-479. https://doi.org/10.1086/376536
- Macdonald F, Odds F. (1983). Virulence for mice of a proteinase-secreting strain of Candida albicans and a proteinase-deficient mutant. Microbiology, 129(2): 431-438. https://doi.org/10.1099/00221287-129-2-431
- Deorukhkar S C, Saini S. (2014). Laboratory approach for diagnosis of candidiasis through ages. International Journal of Current Microbiology and Applied Sciences, 3(1): 206-218.
- Hospenthal D R, Beckius M L, Floyd K L, Horvath L L, Murray C K. (2006). Presumptive identification of Candida species other than C. albicans, C. krusei, and C. tropicalis with the chromogenic medium CHROMagar Candida. Annals of clinical microbiology and antimicrobials, 5(1): 1-5. https://doi.org/10.1186/1476-0711-5-1
- PM T. (2017). Bailey & Scott’s Diagnostic Microbiology, St. Louis (pp. 1056 Pages): Elsevier.
- Van Leekwijck W, Kerre E E. (1999). Defuzzification: criteria and classification. Fuzzy sets and systems, 108(2): 159-178. https://doi.org/10.1016/S0165-0114(97)00337-0
- Pinjon E, Sullivan D, Salkin I, Shanley D, Coleman D. (1998). Simple, inexpensive, reliable method for differentiation of Candida dubliniensis from Candida albicans. Journal of Clinical Microbiology, 36(7): 2093-2095. https://doi.org/10.1128/JCM.36.7.2093-2095.1998
- Abdulla H, Mustafa E A A. (2020). Rapid Detection of Candida species Isolated from Denture Stomatitis Patients using Phenotypic methods and Chromogenic agar media. Al-Rafidain Dental Journal, 20(1): 125-133. http://dx.doi.org/10.33899/rden.2020.126821.1029
- Wibawa T, Aman A T. (2015). Virulence of Candida albicans isolated from HIV infected and non infected individuals. SpringerPlus, 4(1): 1-10. https://doi.org/10.1186/s40064-015-1215-0
- Singh M, Chakraborty A. (2017). Antifungal Drug Resistance among Candida albicans and Non-albicans Candida Species Isolates from a Tertiary Care Centre at Allahabad. J Antimicrob Agents, 3(150): 2472-1212.1000150. https://doi.org/10.4172/2472-1212.1000150
- Mehta A, Kumar M, Bhumbla U, Vyas A, Dalal A. (2018). Comparison of Different Media for Germ Tube Production by Candida albicans: A Retrospective Study. Int. J. Curr. Microbiol. App. Sci, 7(6): 819-823. https://doi.org/10.20546/ijcmas.2018.706.096
- Salah N S, Muhsen T A, Risan M H. (2020). Antifungal Activity of Silver Nanoparticles Using Penicillium Chrysogenum Extract Against The Formation of Biofilm for Candida Glabrata. Indian J. of Forensic Medicine and Toxicology, 14(2): 306-311.
- Hussein H, Dheeb B, Hamada T. (2019). Studying the candida resistance and sensitivity for some antifungals. Journal of Biotechnology Research Center, 13(2): 26-34.
- Wohlmeister D, Vianna D R B, Helfer V E, Calil L N, Buffon A, Fuentefria A M, Corbellini V A, Pilger D A. (2017). Differentiation of Candida albicans, Candida glabrata, and Candida krusei by FT-IR and chemometrics by CHROMagar™ Candida. Journal of Microbiological Methods, 141: 121-125. https://doi.org/10.1016/j.mimet.2017.08.013
- Ghasemi R, Rabiei M M, Lotfali E, Abolghasemi S, Ansari S. (2020). Candiduria: Prevalence, identification of isolated candida species and trends in antifungal susceptibility in hospitalized patients. Novelty in Biomedicine, 8(2): 71-76. https://dx.doi.org/10.22037/nbm.v1i1.26963
- De Backer M D, Ilyina T, Ma X-J, Vandoninck S, Luyten W H, Vanden Bossche H. (2001). Genomic profiling of the response of Candida albicans to itraconazole treatment using a DNA microarray. Antimicrobial agents and chemotherapy, 45(6): 1660-1670. https://doi.org/10.1128/AAC.45.6.1660-1670.2001
- Zhao X, Pujol C, Soll D R, Hoyer L L. (2003). Allelic variation in the contiguous loci encoding Candida albicans ALS5, ALS1 and ALS9. Microbiology, 149(10): 2947-2960. https://doi.org/10.1099/mic.0.26495-0
- Naglik J R, Challacombe S J, Hube B. (2003). Candida albicans secreted aspartyl proteinases in virulence and pathogenesis. Microbiology and Molecular Biology Reviews, 67(3): 400-428. https://doi.org/10.1128/MMBR.67.3.400-428.2003
- Ilkhanizadeh-Qomi M, Nejatbakhsh S, Jahanshiri Z, Razzaghi-Abyaneh M. (2020). Aspartyl Proteinase and Phospholipase Activities of Candida albicans Isolated From Oropharyngeal Candidiasis in Head and Neck Cancer Patients. Jundishapur Journal of Microbiology, 13(9). https://dx.doi.org/10.5812/jjm.105200
- Mothibe J V, Patel M. (2017). Pathogenic characteristics of Candida albicans isolated from oral cavities of denture wearers and cancer patients wearing oral prostheses. Microbial pathogenesis, 110: 128-134. https://doi.org/10.1016/j.micpath.2017.06.036
- Ganesan K, Banerjee A, Datta A. (1991). Molecular cloning of the secretory acid proteinase gene from Candida albicans and its use as a species-specific probe. Infection and immunity, 59(9): 2972-2977. https://doi.org/10.1128/iai.59.9.2972-2977.1991