Oral Candida Overgrowth in most Thalassemia Major Patients, Yeast Identification by CDA, VITEK 2, and PCR, and Evaluating their Virulence Factors in Thi-Qar, Iraq

Pertumbuhan Candida Oral yang Berlebih pada Sebagian Besar Pasien Thalassemia Mayor, Identifikasi Ragi dengan CDA, VITEK 2, dan PCR, dan Evaluasi Faktor Virulensinya di Thi-Qar, Irak

Authors

  • Salih Jabbar Department of Biology, Colleg of Science, University of Basrah, Basrah
  • Furdos Nouri Jafar Department of Biology
  • Shereen Al-Ali Department of Pathological Analyses, College of Science, University of Basrah, Basrah
  • Afaq Hameed Department of Bio-Medical Engineering, College of Engineering, University of ThiˍQar, ThiˍQar

DOI:

https://doi.org/10.21070/ijhsm.v2i1.146

Keywords:

accuracy of Candida identification, biofilm, extracellular enzymes, oral Candida overgrowth, thalassemia major patients

Abstract

Candida, an opportunistic fungus in immunocompromised patients, can cause an infection like oral candidiasis which usually starts with Candida overgrowth. Thalassemia major (TM) patients are known to suffer from immune abnormalities that may lead to a condition of immune deficiency. So, we aimed to estimate the prevalence and intensity of Candida growth in oral cavity of TM patients, determine the accuracy of Candida identification, and assess some virulence factors. Our study was conducted on 150 TM patients and 80 controls between (1-40 years). Oral swabs were used for microscopic examination and culturing on SDA. The identification was done by candida differential agar (CDA), VITEK 2, and conventional PCR. Also, the virulence was estimated by measurement of proteinase, phospholipase, lipase, hemolysin, and biofilm. Candida species were orally isolated from 70% of TM patients with mean colony count (124±94) significantly more than 42.5% of control with (11±7), so a significant oral Candida overgrowth was observed in most (TM) patients compared with the control, referring to an increased probability of developing to oral candidiasis. Significantly, in total TM patients, male or female patients, the age group (11–20) showed a higher prevalence of Candida than other age groups, at 52.4%, 49%, and 55.56%, respectively. PCR identified 105 isolates from TM patients: C. albicans constituted the most common species with (61.9%), C. dubliniensis (35.2%) and C. glabrata (2.86%). Generally, in comparison with results of PCR, the accuracy of identification by CDA was (95.2%) more than (87.6%) by VITEK 2, but both were typical methods for identifying C. albicans with (100%). Significantly, the higher production of proteinase and lipase was by (92.3%) and (90.7%) of C. albicans isolates, respectively. While the majority of phospholipase and biofilm production was noted by (70.3%) of C. dubliniensis and (100%) of C. glabrata, respectively. All Candida species were hemolysin producers with 100%.

 Highlights:

  1. Samples: 150 TM patients, 80 controls; oral Candida prevalence 70%.
  2. Identification: PCR, CDA, VITEK 2; C. albicans most common (61.9%).
  3. Virulence: High proteinase, lipase (C. albicans); biofilm (C. glabrata).

Keywords: accuracy of Candida identification, biofilm, extracellular enzymes, oral Candida overgrowth, thalassemia major patients

References

[1]. G. Moran, D. Coleman, and D. Sullivan, “An introduction to the medically important candida species,” in ASM Press eBooks, 2014, pp. 9–25. doi: 10.1128/9781555817176.ch2.

[2]. A.-C. Bostănaru et al., “Genotype comparison of Candida albicans isolates from different clinical samples,” Revista Română De Medicină De Laborator, vol. 27, no. 3, pp. 327–332, Jul. 2019, doi: 10.2478/rrlm-2019-0027.

[3]. J. W. Millsop and N. Fazel, “Oral candidiasis,” Clinics in Dermatology, vol. 34, no. 4, pp. 487–494, Mar. 2016, doi: 10.1016/j.clindermatol.2016.02.022.

[4]. A. Al-Laaeiby, S. Ali, and A. H. Al-Saadoon, “Candida species: the silent enemy,” African Journal of Clinical and Experimental Microbiology, vol. 20, no. 4, p. 260, Aug. 2019, doi: 10.4314/ajcem.v20i4.1.

[5]. S. Dabiri, M. Shams-Ghahfarokhi, and M. Razzaghi-Abyaneh, “Comparative analysis of proteinase, phospholipase, hydrophobicity and biofilm forming ability in Candida species isolated from clinical specimens,” Journal De Mycologie Médicale, vol. 28, no. 3, pp. 437–442, May 2018, doi: 10.1016/j.mycmed.2018.04.009.

[6]. K. I. Othman, S. M. Abdullah, B. Ali, and M. Majid, “Isolation and Identification Candida spp from Urine and Antifungal Susceptibility Test,” Nov. 28, 2018. https://ijs.uobaghdad.edu.iq/index.php/eijs/article/view/554

[7]. A. Bazi, I. Shahramian, H. Yaghoobi, M. Naderi, and H. Azizi, “The role of Immune System in Thalassemia Major: A Narrative review,” Journal of Pediatrics Review, vol. 6, no. 2, Oct. 2017, doi: 10.5812/jpr.14508.

[8]. A. Gluba-Brzózka, B. Franczyk, M. Rysz-Górzyńska, R. Rokicki, M. Koziarska-Rościszewska, and J. Rysz, “Pathomechanisms of immunological disturbances in Β-Thalassemia,” International Journal of Molecular Sciences, vol. 22, no. 18, p. 9677, Sep. 2021, doi: 10.3390/ijms22189677.

[9]. J. Dwye et al., “Abnormalities in the immune system of children with beta-thalassaemia major,” PubMed, Jun. 01, 1987. https://pubmed.ncbi.nlm.nih.gov/3498580/

[10]. N. M. Abu-Mejdad et al., “A new record of interesting basidiomycetous yeasts from soil in Basrah province/Iraq” Basrah Journal of Science, vol. 3, no. 37, p. 9307-34, Sep. 2019.

[11]. A. I. E. Al-Laaeiby, A. A. Al-Mousawi, I. M. N. Alrubayae, A. Al-Saadoon, and M. Almayahi, “Innate pathogenic traits in oral yeasts,” Karbala International Journal of Modern Science. https://kijoms.uokerbala.edu.iq/home/vol6/iss4/5/

[12]. V. Sunitha, D. Nirmala Devi, C. Srinivas, “Extracellular enzymatic activity of endophytic fungal strains isolated from medicinal plants” World Journal of Agricultural Sciences, vol. 9, no. 1, p. 1-9, 2013.

[13]. G. Luo, L. P. Samaranayake, and J. Y. Y. Yau, “Candida species exhibit differential in vitro hemolytic activities,” Journal of Clinical Microbiology, vol. 39, no. 8, pp. 2971–2974, Aug. 2001, doi: 10.1128/jcm.39.8.2971-2974.2001.

[14]. S. Abdulla, B. Dheeb, S. A.-D. Al-Qaysi, M. S. Farhan, M. Massadeh, and A. Diab, “Isolation and Diagnosis of Biofilms of Klebsiella pneumoniae Bacteria and Candida albicans Yeast, and Studying the Sensitivity of The Pathogens to Antibiotics and Antifungals,” Egyptian Academic Journal of Biological Sciences. G, Microbiology, vol. 15, no. 2, pp. 73–88, Oct. 2023, doi: 10.21608/eajbsg.2023.320271.

[15]. C. Asadov D. and Institute of Hematology and Transfusiology, Baku, Azerbaijan, “Immunologic abnormalities in Β-Thalassemia,” J Blood Disorders Transf, vol. 5, no. 7, p. 224, Jul. 2014, [Online]. Available: https://www.walshmedicalmedia.com/open-access/immunologic-abnormalities-in-thalassemia-2155-9864.1000224.pdf

[16]. C. Thiengtavor et al., “Increased ferritin levels in non‐transfusion‐dependent β°‐thalassaemia/HbE are associated with reduced CXCR2 expression and neutrophil migration,” British Journal of Haematology, vol. 189, no. 1, pp. 187–198, Dec. 2019, doi: 10.1111/bjh.16295.

[17]. W. Wanachiwanawin, “Infections in E-Β thalassemia,” the American Journal of Pediatric Hematology/Oncology, vol. 22, no. 6, pp. 581–587, Nov. 2000, doi: 10.1097/00043426-200011000-00027.

[18]. G. M. Mokhtar, M. Gadallah, N. H. K. E. Sherif, and H. T. A. Ali, “Morbidities and mortality in Transfusion-Dependent Beta-Thalassemia patients (Single-Center experience),” Pediatric Hematology and Oncology, vol. 30, no. 2, pp. 93–103, Jan. 2013, doi: 10.3109/08880018.2012.752054.

[19]. D. Dadhich, N. Saxena, A. E. Chand, G. Soni, and S. Morya, “Detection of candida species by hichrom agar and their antimycotic sensitivity in Hadoti region,” International Journal of Scientific Study, journal-article, Jul. 2016. doi: 10.17354/ijss/2016/367.

[20]. S. Mathavi, G. Sasikala, A. Kavitha, and R. I. Priyadarsini, “CHROMagar as a primary isolation medium for rapid identification of Candida and its role in mixed Candida infection in sputum samples,” www.academia.edu, Jul. 2016,

[21]. B. Graf, T. Adam, E. Zill, and U. B. GöBel, “Evaluation of the VITEK 2 System for Rapid Identification of Yeasts and Yeast-Like Organisms,” Journal of Clinical Microbiology, vol. 38, no. 5, pp. 1782–1785, May 2000, doi: 10.1128/jcm.38.5.1782-1785.2000.

[22]. E. Aboualigalehdari, M. T. Birgani, M. Fatahinia, and M. Hosseinzadeh, “Oral Colonization by Candida Species and Associated Factors among HIV-infected Patients in Ahvaz, Southwest Iran,” Epidemiology and Health, p. e2020033, May 2020, doi: 10.4178/epih.e2020033.

[23]. M. A. Kabir, M. A. Hussain, and Z. Ahmad, “Candida albicans: A Model Organism for Studying Fungal Pathogens,” ISRN Microbiology, vol. 2012, pp. 1–15, Sep. 2012, doi: 10.5402/2012/538694.

[24]. B. Böttcher, C. Pöllath, P. Staib, B. Hube, and S. Brunke, “Candida species Rewired Hyphae Developmental Programs for Chlamydospore Formation,” Frontiers in Microbiology, vol. 7, Oct. 2016, doi: 10.3389/fmicb.2016.01697.

[25]. R. Fourie, O. O. Kuloyo, B. M. Mochochoko, J. Albertyn, and C. H. Pohl, “Iron at the Centre of Candida albicans Interactions,” Frontiers in Cellular and Infection Microbiology, vol. 8, Jun. 2018, doi: 10.3389/fcimb.2018.00185.

[26]. F. C. Odds and R. Bernaerts, “CHROMagar Candida, a new differential isolation medium for presumptive identification of clinically important Candida species,” Journal of Clinical Microbiology, vol. 32, no. 8, pp. 1923–1929, Aug. 1994, doi: 10.1128/jcm.32.8.1923-1929.1994.

[27]. N. V. Jose, N. Mudhigeti, J. Asir, S. D. Chandrakesan, “Detection of virulence factors and phenotypic characterization of Candida isolates from clinical specimens”. Journal of Current Research in Scientific Medicine, vol. 1, no. 1, pp. 27, 2015.

[28]. N. Abu-Mejdad, A. I. Al-Badran, “Isolation and molecular identification of yeasts, study their potential for producing killer toxins and evaluation of toxins activity against some bacteria and pathogenic fungi” Basrah: PhD thesis. PP. 1-249, 2019.

[29]. D. J. Hata, L. Hall, A. W. Fothergill, D. H. Larone, and N. L. Wengenack, “Multicenter evaluation of the new VITEK 2 Advanced Colorimetric Yeast Identification Card,” Journal of Clinical Microbiology, vol. 45, no. 4, pp. 1087–1092, Feb. 2007, doi: 10.1128/jcm.01754-06.

[30]. M. Kord et al., “Epidemiology of yeast species causing bloodstream infection in Tehran, Iran (2015–2017); superiority of 21-plex PCR over the Vitek 2 system for yeast identification,” Journal of Medical Microbiology, vol. 69, no. 5, pp. 712–720, May 2020, doi: 10.1099/jmm.0.001189.

[31]. T. Peremalo et al., “Antifungal susceptibilities, biofilms, phospholipase and proteinase activities in the Candida rugosa complex and Candida pararugosa isolated from tertiary teaching hospitals,” Journal of Medical Microbiology, vol. 68, no. 3, pp. 346–354, Feb. 2019, doi: 10.1099/jmm.0.000940.

[32]. P. Czechowicz, J. Nowicka, and G. Gościniak, “Virulence Factors of Candida spp. and Host Immune Response Important in the Pathogenesis of Vulvovaginal Candidiasis,” International Journal of Molecular Sciences, vol. 23, no. 11, p. 5895, May 2022, doi: 10.3390/ijms23115895.

[33]. W. T. H. Tanaka, N. Nakao, T. Mikami, and T. Matsumoto, “Hemoglobin Is Utilized byCandida albicansin the Hyphal Form but Not Yeast Form,” Biochemical and Biophysical Research Communications, vol. 232, no. 2, pp. 350–353, Mar. 1997, doi: 10.1006/bbrc.1997.6247.

[34]. C. R. Costa, X. S. Passos, L. K. H. E. Souza, P. De Andrade Lucena, O. De Fátima Lisboa Fernandes, and M. D. R. R. Silva, “Differences in exoenzyme production and adherence ability of Candida spp. isolates from catheter, blood and oral cavity,” Revista Do Instituto De Medicina Tropical De São Paulo, vol. 52, no. 3, pp. 139–143, Jun. 2010, doi: 10.1590/s0036-46652010000300005.

[35]. M. Mroczyńska and A. Brillowska-Dąbrowska, “Virulence of clinical candida isolates,” Pathogens, vol. 10, no. 4, p. 466, Apr. 2021, doi: 10.3390/pathogens10040466.

[36]. C. Sachin, K. Ruchi, and S. Santosh, “In vitro evaluation of proteinase, phospholipase and haemolysin activities of Candida species isolated from clinical specimens,” 2012. https://www.ajol.info/index.php/ijmbr/article/view/91862

[37]. N. Ramesh, M. Priyadharsini, C. S. Sumathi, V. Balasubramanian, J. Hemapriya, and R. Kannan, “Virulence Factors and Anti Fungal Sensitivity Pattern of Candida Sp. Isolated from HIV and TB Patients,” Indian Journal of Microbiology, vol. 51, no. 3, pp. 273–278, Apr. 2011, doi: 10.1007/s12088-011-0177-3.

[38]. D. A. Schofield, C. Westwater, T. Warner, and E. Balish, “DifferentialCandida albicanslipase gene expression during alimentary tract colonization and infection,” FEMS Microbiology Letters, vol. 244, no. 2, pp. 359–365, Feb. 2005, doi: 10.1016/j.femsle.2005.02.015.

[39]. R. Alves, et al., Adapting to survive: “How Candida overcomes host-imposed constraints during human colonization”, PLoS pathogens. Vol. 5, no. 16, pp. 1008478, 2020.

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Published

2025-05-01

How to Cite

Jabbar, S., Jafar, F. N., Al-Ali, S., & Hameed, A. (2025). Oral Candida Overgrowth in most Thalassemia Major Patients, Yeast Identification by CDA, VITEK 2, and PCR, and Evaluating their Virulence Factors in Thi-Qar, Iraq: Pertumbuhan Candida Oral yang Berlebih pada Sebagian Besar Pasien Thalassemia Mayor, Identifikasi Ragi dengan CDA, VITEK 2, dan PCR, dan Evaluasi Faktor Virulensinya di Thi-Qar, Irak. Indonesian Journal on Health Science and Medicine, 2(1), 10.21070/ijhsm.v2i1.146. https://doi.org/10.21070/ijhsm.v2i1.146

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