Users Online: 900
Home Print this page Email this page
Home About us Editorial board Search Browse articles Submit article Ahead of Print Instructions Subscribe Contacts Login 


 
Previous article Browse articles Next article 
ORIGINAL ARTICLE
Adv Biomed Res 2022,  11:29

Prevalence, molecular identification, and genotyping of Candida species recovered from oral cavity among patients with diabetes mellitus from Tehran, Iran


1 Department of Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
2 Department of Parasitology and Mycology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
3 Department of General Microbiology, Microbiology Institute, Federal University of Rio de Janeiro, Brazil
4 Infectious Disease Research Center, Birjand University of Medical Sciences, Birjand, Iran
5 Department of Medical Parasitology and Mycology, Infectious Diseases and Tropical Medicine Research Center, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

Date of Submission08-Feb-2021
Date of Acceptance15-Nov-2021
Date of Web Publication29-Apr-2022

Correspondence Address:
Dr. Shahla Roudbar Mohammadi
Department of Mycology, Faculty of Medical Science, Tarbiat Modares University, Tehran
Iran
Login to access the Email id

Source of Support: None, Conflict of Interest: None


DOI: 10.4103/abr.abr_26_21

Rights and Permissions
  Abstract 


Background: Oral candidiasis (OC) has been noticed as a common mucous membrane infection in immunocompromised patients such as that diabetes. This study, focused on the genotyping of Candida albicans and enzymatic activities of Candida species recovered from oral mucosa among diabetes patients and healthy individuals. Materials and Methods: Specimens were obtained from oral mucosa of One-hundred and sixty patients with type 2 diabetic and 108 healthy individuals. All isolates were definitely identified by ribosomal DNA (rDNA) gene sequencinghHydrophobicity, hemolytic activities of Candida species and genotypes of C. albicans were determined through polymerase chain reaction (CA-INT). Results:, Eighty eight (55%) samples out of 160, were positive for Candida species in diabetic patients. Moreover, 79.5% (70/88) and 20.5% (18/88) isolates belonged to the C. albicans and non-albicans Candida species respectively. Three genotypes of C. albicans have recovered in diabetic patients: genotype A (71.42%), B (21.42%), and C (7.14%). In healthy individuals, 42.6% (46/102) Candida species recovered from oral cavity, with the highest prevalence of genotype A (76.6% of C. albicans). Additionally, hydrophobicity and hemolytic activities from Candida species were significantly greater in diabetes patients than healthy nondiabetic subjects. Conclusion: Collectively, C. albicans was the most causative agent isolated from diabetes patients and non-diabetes healthy individuals. Genotype A, as the most remarkable genotype, should be mentioned in both groups. Higher potential hydrophobicity and hemolytic activities of Candida species in diabetic patients compared to healthy cases suggest these features triggering pathogenicity of OC in diabetes patients.

Keywords: Candida species, Candidiasis, Diabetes mellitus, Genotype


How to cite this article:
Zarei N, Roudbary M, Roudbar Mohammadi S, dos Santos AL, Nikoomanesh F, Mohammadi R, Shirvan BN, Yaalimadad S. Prevalence, molecular identification, and genotyping of Candida species recovered from oral cavity among patients with diabetes mellitus from Tehran, Iran. Adv Biomed Res 2022;11:29

How to cite this URL:
Zarei N, Roudbary M, Roudbar Mohammadi S, dos Santos AL, Nikoomanesh F, Mohammadi R, Shirvan BN, Yaalimadad S. Prevalence, molecular identification, and genotyping of Candida species recovered from oral cavity among patients with diabetes mellitus from Tehran, Iran. Adv Biomed Res [serial online] 2022 [cited 2022 May 23];11:29. Available from: https://www.advbiores.net/text.asp?2022/11/1/29/344359




  Introduction Top


In recent decades, the rate of opportunistic fungal infections is incredibly increasing worldwide. Accordingly, among them, oral candidiasis (OC) is one of the most common mucous membrane complications in people with the impaired immune system.[1]

Candida albicans (C. albicans) is a normal microbiota of the skin and mucous membranes causing a wide range of superficial infections to fatal systemic infections, especially among immunocompromised subjects such as patients with HIV/AIDS, diabetes, hospitalized subjects receiving long-term antibiotics, patients under organ transplant, chemotherapy or radiotherapy.[2],[3],[4] Significantly, diabetes causes one of the largest emerging threats to public health in the 21st century.[5],[6] Correspondingly, the World Health Organization and the International Diabetes Federation predicted that the number of adults with diabetes will rise near 629 million by the year 2045 in worldwide.[7] Approximately 4.5 million adult people were living with diabetes in Iran in 2011, it is expected that by the year 2030, 9.2 million Iranian persons will have diabetes. This continuous and noteworthy rise in the prevalence of diabetes reveals the high burden of disease in Iran, particularly diabetes-associated problems.[8]

Previous reports have widely demonstrated that the rate of Candida carriage rises in diabetic subjects suffering from oral infections compared to non-diabetic individuals.[9],[10] A high glucose level in salivary, uncontrolled hyperglycemia,[11],[12],[13],[14] lower salivary pH,[14],[15] oral epithelium alteration,[16] and impaired neutrophils' response[16],[17],[18] are associated with physiopathology and etiology of occurrence of OC in diabetic patients. Notably, C. albicans was the most common species isolated from patients; however, the prevalence of non-albicans Candida species (NACs) are increasing dramatically.[19],[20]

Various potential properties of Candida species especially cell surface hydrophobicity (CSH) accelerate the adherence of Candida to host mucous membranes of oral cavity. Adhesion to the tissue resulting in biofilm formation, disperse of the infection, invasion, and finally leading to emerging antifungal resistant species. As a result, Candida adherence to epithelial cells of the oral cavity is a key initial step in the pathogenicity of OC infection.[21],[22],[23] Surprisingly, enzymatic activity, particularly hemolytic ability of Candida species, gives an opportunity to acquire iron of host tissue and to enhance the virulence toward the progression of infection.[24],[25],[26]

To realize the source of infection, the identification of a genetic link between strains based on typing methods is highly noticed.[27] For this purpose, a large number of molecular typing methods such as random amplified polymorphic DNA (RAPD) fingerprinting,[20] multilocus sequence typing (MLST),[28] electrophoretic karyotyping,[29] and ABC genotyping,[30] with the matter technique based on the presence or absence of an intron in the 25S ribosomal DNA (rDNA), are used to determine the typing of clinical isolates of Candida strains. In Iran as a developing country, improvement and use of rapid, cost-benefit and reliable methods for genotyping of fungal isolates attract many attentions due to the limited available sources.

Although previous studies have reported the prevalence of Candida species in oral mucosa of diabetic patients in different geographic regions of Iran, genotyping pattern of isolates is not well-studied yet.[31],[32],[33] To address this question, in the current study, we isolated and identified the Candida species of oral cavity of both diabetic patients and healthy nondiabetic subjects, which have mainly focused on the determination of genotyping of C. albicans (25S rDNA region) through polymerase chain reaction (PCR) (CA-INT) as well as on the production of enzymatic activities of Candida species.


  Materials and Methods Top


Clinical specimens, culture condition

In the period (October 2018 to November 2019), 160 patients with type 2 diabetes, who were attended at the Diabetic centers (Tehran, Iran), have been recruited in this cross-section study. Besides, 108 non-diabetic individuals were also included in this study. All the participants had assigned a consent form at the beginning of sampling and subsequently, they were identified by numerical codes. All demographic data of patients including age, gender, clinical sign of OC, and hemoglobin A1c (HbA1c) was recorded in questionnaires' form.

Sterile cotton swab samples were taken from the tongue and buccal mucosal lesions of patients, which were immediately transported to Falcone contains sterile phosphate buffer saline (PBS). All specimens were tested for direct microscopic examination, to detect probable yeast and pseudohyphae elements. Subsequently, specimens were cultured on Sabouraud dextrose agar medium (SDA, Sigma Aldrich-USA) containing chloramphenicol, and then incubated for 24–48 h at 35°C–37°C. Moreover, Candida isolates were presumptively identified using chromogenic medium (CHROMagarCandida, France), as well as the production of chlamydospores on cornmeal agar for morphological identification. In addition, all the fungal isolates were definitely confirmed by PCR and sequencing as described below.

Identification of Candida species, DNA amplification, and sequencing

Genomic DNA was extracted from Candida species using glass beads and lysis buffer as described earlier.[34],[35] Finally, for the precise identification of Candida species, PCR-sequencing of the internal transcribed spacer (ITS) region (ITS 1-5.8s-ITS2 rDNA region) was carried out and the obtained data were then analyzed in NCBI gene bank and blasted based on similarity of Nucleotide Blast database. Universal primers were also used for the ITS gene related to fungal rDNA genes. Nucleotide sequences of Candida species are available on NCBI website.

Genotyping of Candida albicans

Genomic DNA of C. albicans isolates was extracted as previously described with minor modification.[34] Then, the quality of DNA was checked out using electrophoresis on agarose gel 1.5%. Subsequently, PCR assay was performed as the following process: 12.5 μl of Master mix PCR (Amplicone, Korea) was added to 10 picomole from each specific primers (Forward 5'-CCT TGG CTG TGG TTT CGC TAG ATA GTA GAT-3') and (Reveres5'-ATA AGG GAA GTC GGC AAA ATA GAT CCG TAA-3'), then 2 μl DNA from each isolate was added to the mixture, and the final volume adjusted to 25 μl by distilled water. Afterward, the tube was placed in a Thermal Cycler (PreQLab, Germany). PCR program was adjusted 30 cycles containing denaturation for 5 min at 95°C, annealing for 1 min at 65°C, extension for 1 min at 72°C, and the final extension for 7 min at 95°C. Subsequently, the amplified products were analyzed by electrophoresing on 1.5% gel agarose in TAE (Trisbase, acetic acid, EDTA, Sigma-Aldrich, USA) buffer and then stained with 0.5 μg/ml SYBR safe-DNA Gel stain (Takara, Japan), to visualize the bands. Based on the molecular size of different genotypes, we categorized them into three groups as follows: genotype A exhibiting one band size of 450 base pair (bp), genotype representing one band size of 650 bp, and genotype C exhibiting two band sizes of 450 and 650 bp.[36]

Hemolytic assay

Hemolytic activity of Candida isolates was evaluated using a modification of the plate assay described by Sardi et al.[29] Briefly, the isolates were cultured on Sabouraud dextrose agar (Merck, Germany) containing chloramphenicol for 24 hat 35°C–37°C. In the next step, Sabourauddextrose agar medium, which was enriched with fresh human blood red cells (RBC) and glucose in the final concentration 3% w/v, was prepared. For this purpose, small wells (size 2–3 mm) were created in the medium, and 10 μL equal to 1.5 × 103 yeasts per ml of fresh yeast isolates was individually grown on SDA and then inoculated onto each well of blood agar. Notably, a reference strain of C. albicans (ATCC 90028) was used as a positive control. In addition, one standard strain of Staphylococcus aureus and one strain of Streptococcus mutans, inducing beta and alpha hemolysis were used as positive controls, respectively. Then, the plates were incubated for 48 hat 37°C in 5% CO2. Alpha-hemolytic (α-hemolytic) strains produced a green halo and beta-hemolytic (β-hemolytic) strains produced a yellow halo. The experiments were performed in triplicate in three independent experiments for each one of the tested yeast isolates. Next, hemolysis and/or non-hemolysis caused by isolates were measured using the following formula: areola diameter = total circle diameter created-diameter of the hole created in the plate.

Hydrophobicity assay

Candida isolates were cultured on SDA medium and then incubated for 24 hat 35°C–37°C. Yeast suspension was prepared (1.5 × 103 yeast per ml), transferred to microtube containing 0.5 ml sterile YNB medium (yeast nitrogen base, Sigma-Aldrich, USA) for 24 h. Then, YNB medium was discarded and the sediment was then transferred to micro tubes containing 1 ml sterile PBS. Optical density (OD) of the samples was measured using spectra photometer with a wavelength of 520 nm. Subsequently, n-octan 25 (TITRACHEM company) was added to yeast suspension and this mixture was incubated for 30 min at 37°C and finally, two phases were observed. Subsequently, the upper phase was discarded and OD of lower-phase was measured for hydrophobicity activity. The OD values of strains in the YNB broth without n-octane overlay were used as negative controls. In addition, the relative CSH was calculated according to the following formula: [(OD570 of the control-OD570 after n-octane overlay)/OD570 of the control] x 100. The results were expressed as a percentage (%).[21]

Statistical analysis

The data were first analyzed using SPSS software, version 20 (SPSS, Chicago, IL, USA), and subsequently analyzed using descriptive statistics, Fisher's, and Pearson Chi-square tests to determine differences in the ratio of the categorized variables. Afterward, continuous variables with an approximately normal distribution were tested using the Student's test. Furthermore, P < 0.05 was considered as statistically significant.


  Results Top


Patients'data, Candida species, and enzymatic activity

Of 160 diabetic patients, 69 (43.12%) were men (mean age of 65 years old) and 91 (56.87%) were women (mean age of 68 years old) while out of 108 healthy subjects, (n = 17, 16%) were men (mean age of 60 years old) and (n = 91, 84%) were women (mean age of 64 years old). The mean HbA1c of the diabetes patients was determined to be 9% and 9.5% in men and women, respectively (Normal range of HbA1c is below 6%) whereas the mean of HbA1c level was determined as 4% for both of gender. The demographic data of the patients are summarized in [Table 1]. Thrush was the most frequent clinical signs of oral lesions in the diabetic patients (n = 50, 56.8%) followed by pseudomembranous plaques (n = 20, 22.7%) and perleche (n = 18, 20.5%).
Table 1: Demographic characteristic, enzymatic activity and genotyping of Candida species in diabetic patients compared to health subjects

Click here to view


A total of 88 (55%) patients with diabetes were positive for Candida species on SDA medium. Using CHROMagar Candida medium, 79.5% of the isolates (70/88) were presumptively identified as C. albicans, whereas 18/88 (20.5%) isolates were identified as NACs. Totally, 46 (42.6%) Candida species recovered from healthy individuals, among the positive species, 43 (93.5%) were identified as C. albicans, and only (n = 3, 6.5%) were NACs, including C. glabrata (n = 2) and Candida dublinensis (n = 1) [Table 2].
Table 2: Frequency of Candida species identified from oral cavity of diabetes patients and control group

Click here to view


Subsequently, all the Candida isolates were definitely identified by sequencing of ITS 1-5.8s-ITS2 rDNA region and then matched to the reference documents in the GenBank database using BLAST to distinguish species' level. The accession numbers of Candida species are represented in (http://www.ncbi.nlm.nih.gov/BLAST) as follows: MG821633, MG821632.1, MG821630.1, MG821628.1, MG821625.1, MG812312.1, KY773288, and MG812310.

In diabetes patients, C. albicans (n = 70, 79.5%) was the most frequent species followed by NACs: Candida krusei (n = 10; 11.4%), C. dublinensis (n = 5; 5.7%), and C. kefyr (n = 3; 3.4%) [Table 2].

In patients with diabetes, the majority of Candida species (64/88, 72.72%) presented hydrophobicity >88% (high hydrophobicity is considered above 60%). In healthy non-diabetes subjects, hydrophobicity of C. albicans was 60%. Remarkably, the hydrophobicity of Candida species was significantly greater in patients with diabetes compared to the control group (P = 0.03). Similarly, in diabetic patients, C. albicans species showed a significantly higher level of hydrophobicity compared to NACs (P < 0.05) isolates.

In diabetes patients, hemolytic activities of the majority of both C. albicans (82.1%) and NACs (17.9%) were more than 19.5%, whereas in healthy individuals, hemolytic activity of C. albicans was detected 10%.(P = 0.04). Interestingly, a statistically significant difference was observed in terms of hemolytic activity of Candida species between the healthy controls and diabetes patients (P < 0.05). However, no significant association was detected between hemolytic activity and Candida species in both diabetic patients and healthy subjects (P = 0.56).

Genotyping of Candida albicans isolates

Genotyping pattern of C. albicans were determined using amplification of 25S rDNA region through PCR (CA-INT). Three major genotyping patterns were detected in C. albicans isolates recovered from diabetic cases as follows: genotype A was the most predominant one among the isolates (71.42%, n = 50) followed by genotype B (21.43%, n = 15) and genotype C (7.15%, n = 5). No statistically significant association was found between different genotype and hemolytic and hydrophobicity activities of Candida species (P = 0.7). In addition, no statistically significant correlation was observed among genotyping patterns and age and HbA1c level (P = 0.7) in diabetes patients. Genotype A of C. albicans was detected in 26% of diabetic women compared to diabetic men (10%). In healthy controls among C. albicans species (n = 43), genotype A (n = 33, 76.7%) was frequently prevalent followed by genotype B (n = 8, 18.6%), and genotype C (n = 2, 4.7%). The frequency of genotypes of C. albicans in patients with diabetes compared to healthy individuals is shown in [Figure 1]. PCR pattern of different genotypes of gel electrophoresis is shown in [Figure 2].
Figure 1: Frequency of genotyping patterns of Candida albicans isolates from diabetic patients and healthy, nondiabetes individuals

Click here to view
Figure 2: Genotypes of Candida albicans representing different product size of CA-INT amplification on agarose gel electrophoresis. Line 1, 2, 3 and 6,7 genotype A (450 bp), N; Negative control, Line 4 and 5 genotype B (650 bp), M; DNA Marker100 bp

Click here to view



  Discussion Top


The obtained findings showed that the Candida carriage rate was statistically higher in patients with diabetes than healthy individuals, which is in agreement with previous reports.[33],[37] It is well known that immunosuppression condition such diabetes mellitus as a major health challenging, can affect Candida colonization in a high rate and consequently progress the infection in host tissue.[1],[9]

In our study, C. albicans was the most predominantly species of oral cavity in the two groups followed by NACs, which have consistently been studied previously.[33],[38],[39]

Remarkably, C. krusei was the most frequently NACs isolated from oral cavity of diabetes patients. It is noteworthy that the number of NACs has significantly increased over the last decades accompanied by relevant growth in morbidity and mortality rates.[40]

Our finding indicated that hydrophobicity of Candida species was significantly higher in diabetes patients compared to healthy individuals. Moreover, hydrophobicity ability of C. albicans species was significantly greater than NACs. Surprisingly, previous reports support our findings, demonstrating that C. albicans through CSH adhered to epithelial surface of host tissue such as oral mucosa causing enhancing virulence and pathogenesis by increasing the adhesion and prompting biofilm formation in infected tissue.[41],[42],[43] On the other hand, previous studies indicated a high concentration of glucose in individuals with diabetes serve as a nutritional source to maintain the polysaccharide matrix of Candida biofilm to protect this structure from environmental changes.[44]

Biofilm formation[44],[45] and enzymatic activity,[25],[46],[47] serve as two important features of pathogenic ability of Candia species. These factors have a main impact on the balance between host and Candida sp., favoring the transition of Candida from commensal to pathogen and leading to infection.[1] Adherence of Candida in oral mucosa can also exhibit antifungal resistance significantly and persistent infection could reach deep tissue of patients and causing disseminated and life threatens candidiasis like Candidemia in immunosuppressive patients with damaged cellular immunity.[45],[48] Therefore, appropriate strategies to the management of OC in the high-risk patients seems fundamental to control of disseminated infections.

Hemolytic analysis of Candida species revealed a statistically significant difference between patients with diabetes and nondiabetic healthy participants. Unlikely, this activity showed no significant changes between C. albicans and NACs in the two studied group. Studies conducted on the activity of hemolysis in Candida species are limited. Fatahinia et al. demonstrated significantly different hemolytic activity of Candida species isolated from oral mucosa of diabetes patients and healthy, nondiabetic controls.[25] In another study, all C. albicans strains that were isolated from oral cavity of HIV-infected patients produced hemolysins, as a virulence factor, while 86% NACs revealed hemolytic activity.[49]

It was previously shown that Candida isolates among patients with diabetes due to a higher blood glucose concentration, represented a higher pathogenic potential for adhesion and hemolytic activity, which may usually contribute to pathogenesis positively.[46],[50],[51] In another study by Tsang, significantly higher hemolytic activity was proven in Candida species from oral cavity of type 2 diabetes patients compared to the control group.[47] This finding has been supported by the higher blood glucose concentration and HbA1c in patients with diabetes. Hence, Luo et al. defined that the increased glucose concentration in culture medium directly or indirectly enhance hemolysin activity of Candida species isolated from diabetic patients.[52]

According to our findings, genotype A was the most common genotypes among C. albicas species isolated from both diabetes patients and healthy individuals. In this context, we hypothesized that pathogenicity of OC in patients with diabetes is associated with genotype A, as a predominant type. Nevertheless, genotypes B and C with less frequency increased in C. albicans species and must be noted in the diabetic patients. Several factors including gene mutations may contribute to the diversity of genotyping patterns in our study.[53],[54],[55] In spite of genotype A was the most frequent genotype in two populations, no significant association between genotype A, hydrophobicity, and hemolytic of C. albicans species were observed in our study. Sardietal. determined the genetic pattern of C. albicans isolated from diabetic patients with chronic periodontitis. The findings indicated two main genotypes A and B in C. albicans isolates, which was in contrast with our study, and they also reported a positive association between genotyping A and a higher hydrophobicity in Candida isolates.[29] In consistent with our finding, Bii et al.[36] demonstrated different genotypes of C. albicans isolated from blood, sputum, urine, and catheters using primer pairs, which span the site of transposable intron 25S rDNA with the highest frequency of genotype A followed by genotypes B, C, and BC. In a study by Menezes et al., different genotypes of Candida species of the hospitalized patients have been indicated in Brazil using RAPD-PCR.[53]

It seems that strains from different geographical regions have been known to display slight differences in genotypic features.

In conclusion, we reported three genotypes patterns of C. albicans isolates from oral cavity of patients with diabetes and healthy individuals in Iran using Amplification of 25S rDNA region. Notably, the low number of isolates was one of the limitations of this study. By selecting large sample size, we might be able to obtain the possible association between different clinical parameters and virulence factors of Candida species. Amplification of 25S rDNA region through PCR (CA-INT) suggests high levels of reproducibility and discrimination, which facilitate the description of C. albicans genotypes while being easily performed in a laboratory.[56] Definitely, the application of the other confirmatory molecular methods such as Saturn Blatting Hybridization, MLST, and DNA microsatellite analysis would be desired.[29],[57]

Further studies on the various extracellular enzymes responsible for virulence are urgently needed to uncover host-pathogen interaction in connection with mucosal Candida infections in different types of immunosuppressive populations.

Acknowledgments

The authors gratefully acknowledge the Diabetes Clinic of Tehran to provide authors with the samples for this study.

Ethical approval

This study was approved by the research committee of Iran University of Medical Sciences with ethical code (IR. IUMS. REC.573).

Financial support and sponsorship

This study was financially supported by Tarbiat Modares University and Iran University of Medical Sciences with grant number(≠15557).

Conflicts of interest

There are no conflicts of interest.



 
  References Top

1.
Fanello S, Bouchara J, Sauteron M, Delbos V, Parot E, et al. Predictive value of oral colonization by Candida yeasts for the onset of a nosocomial infection in elderly hospitalized patients. J Med Microbiol 2006;55:223-8.  Back to cited text no. 1
    
2.
Rodrigues CF, Rodrigues ME, Henriques M. Candida sp. infections in patients with diabetes mellitus. J Clin Med 2019;8:76.  Back to cited text no. 2
    
3.
Maheshwari M, Kaur R, Chadha S. Candida species prevalence profile in HIV seropositive patients from a major tertiary care hospital in New Delhi, India. J Pathog 2016;2016:6204804.  Back to cited text no. 3
    
4.
Perlroth J, Choi B, Spellberg B. Nosocomial fungal infections: Epidemiology, diagnosis, and treatment. Med Mycol 2007;45:321-46.  Back to cited text no. 4
    
5.
Willis A, Coulter W, Fulton C, Hayes J, Bell P, et al. Oral candidal carriage and infection in insulin-treated diabetic patients. Diabetic Med 1999;16:675-9.  Back to cited text no. 5
    
6.
Karaa A, Goldstein A. The spectrum of clinical presentation, diagnosis, and management of mitochondrial forms of diabetes. Pediatr Diabetes 2015;16:1-9.  Back to cited text no. 6
    
7.
King H, Aubert RE, Herman WH. Global burden of diabetes, 1995-2025: Prevalence, numerical estimates, and projections. Diabetes Care 1998;21:1414-31.  Back to cited text no. 7
    
8.
Esteghamati A, Larijani B, Aghajani MH, Ghaemi F, Kermanchi J, Shahrami A, et al. Diabetes in Iran: Prospective analysis from first nationwide diabetes report of national program for prevention and control of diabetes (NPPCD-2016). Sci Rep 2017;17:13461.  Back to cited text no. 8
    
9.
Kumar B, Padshetty N, Bai KY, Rao M. Prevalence of Candida in the oral cavity of diabetic subjects. J Assoc Phys India 2005;53:599.  Back to cited text no. 9
    
10.
Javed F, Ahmed HB, Mehmood A, Saeed A, Al-Hezaimi K, et al. Association between glycemic status and oral Candida carriage in patients with prediabetes. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2014;117:53-8.  Back to cited text no. 10
    
11.
Rodrigues CF, Henriques. Oral mucositis caused by Candida glabrata biofilms: Failure of the concomitant use of fluconazole and ascorbic acid. Ther Adv Infect Dis 2017;4:10-7.  Back to cited text no. 11
    
12.
Darwazeh A, MacFarlane TW, McCuish A, Lamey PJ. Mixed salivary glucose levels and candidal carriage in patients with diabetes mellitus. J Oral Maxillofac Pathol 1991;20:280-3.  Back to cited text no. 12
    
13.
Naik R, Ahmed Mujib B, Raaju U, Telagi N. Assessing oral candidal carriage with mixed salivary glucose levels as non-invasive diagnostic tool in type-2 diabetics of Davangere, Karnataka, India. J Clin Diagn Res 2014;8:ZC69-72.  Back to cited text no. 13
    
14.
Sashikumar R, Kannan R. Salivary glucose levels and oral candidal carriage in type II diabetics. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010;109:706-11.  Back to cited text no. 14
    
15.
Balan P, Gogineni SB, Kumari S, Shetty V, Rangare AL, et al. Candida carriage rate and growth characteristics of saliva in diabetes mellitus patients: A case-control study. J Dent Res Dent Clin Dent Prospects 2015;9:274-9.  Back to cited text no. 15
    
16.
Malic S, Hill KE, Ralphs JR, Hayes A, Thomas DW, et al. Characterization of Candida albicans infection of an in vitro oral epithelial model using confocal laser scanning microscopy. Oral Microbiol Immunol 2007;22:188-94.  Back to cited text no. 16
    
17.
Martorano-Fernandes L, Dornelas-Figueira LM, Marcello-Machado RM, Silva RB, Magno MB. Oral candidiasis and denture stomatitis in diabetic patients: Systematic review and meta-analysis. Braz Oral Res 2020;34:e113.  Back to cited text no. 17
    
18.
van der Meer JW, van de Veerdonk FL, Joosten LA, Kullberg BJ, Netea MG. Severe Candida spp. infections: New insights into natural immunity. Int J Antimicrob Agents 2010;36:S58-62.  Back to cited text no. 18
    
19.
Zahir R, HimratulAznita WH. Distribution of Candida in the oral cavity and its differentiation based on the internally transcribed spacer (ITS) regions of rDNA. Yeast 2013;30:13-23.  Back to cited text no. 19
    
20.
Barros LM, Boriollo MF, Alves AC, Klein MI, Gonçalves RB, et al. Genetic diversity and exoenzyme activities of Candida albicans and Candida dubliniensis isolated from the oral cavity of Brazilian periodontal patients. Arch Oral Biol 2008;53:1172-8.  Back to cited text no. 20
    
21.
Bujdáková H, Didiášová M, Drahovská H, Černáková L. Role of cell surface hydrophobicity in Candida albicans biofilm. Open Life Sci 2013;8:259-62.  Back to cited text no. 21
    
22.
Singleton DR, Masuoka J, Hazen KC. Surface hydrophobicity changes of two Candida albicans serotype B mnn4delta mutants. Eukaryot Cell 2005;4:639-48.  Back to cited text no. 22
    
23.
Deorukhkar SC, Saini S, Mathew S. Virulence factors contributing to pathogenicity of Candida tropicalis and its antifungal susceptibility profile. Int J Microbiol Res 2014;2014:456878.  Back to cited text no. 23
    
24.
de Melo Riceto ÉB, de Paula Menezes R, Penatti MPA, dos Santos Pedroso R. Enzymatic and hemolytic activity in different Candida species. Rev Iberoam Micol 2015;32:79-82.  Back to cited text no. 24
    
25.
Fatahinia M, Poormohamadi F, Mahmoudabadi AZ. Comparative study of esterase and hemolytic activities in clinically important Candida species, isolated from oral cavity of diabetic and non-diabetic individuals. Jundishapur J Microbiol 2015;8:e20893.  Back to cited text no. 25
    
26.
Chen KW, Chen YC, Lo HJ, Odds FC, Wang TH, et al. Multilocus sequence typing for analyses of clonality of Candida albicans strains in Taiwan. J Clin Microbiol 2006;44:2172-8.  Back to cited text no. 26
    
27.
Colombo AL, Guimarães T, Camargo LF, Richtmann R, Queiroz-Telles FD, et al. Brazilian guidelines for the management of candidiasis-a joint meeting report of three medical societies: Sociedade Brasileira de Infectologia, Sociedade Paulista de Infectologia and Sociedade Brasileira de Medicina Tropical. Braz J Infect Dis 2013;17:283-312.  Back to cited text no. 27
    
28.
Odds FC, Jacobsen MD. Multilocus sequence typing of pathogenic Candida species. Eukaryot Cell 2008;7:1075-84.  Back to cited text no. 28
    
29.
Sardi JC, Duque C, Höfling JF, Gonçalves RB. Genetic and phenotypic evaluation of Candida albicans strains isolated from subgingival biofilm of diabetic patients with chronic periodontitis. Sabouraudia 2012;50:467-75.  Back to cited text no. 29
    
30.
Song X, Eribe ER, Sun J, Hansen BF, Olsen I. Genetic relatedness of oral yeasts within and between patients with marginal periodontitis and subjects with oral health. J Periodent Res 2005;40:446-52.  Back to cited text no. 30
    
31.
Kamali M, Sarvtin MT, Parsanasab H. Prevalence of Candida infection in patients with Type 2 diabetes mellitus in Sari, North of Iran. Biomed Pharmacol J 2016;9:731-4.  Back to cited text no. 31
    
32.
Falahati M, Farahyar S, Akhlaghi L, Mahmoudi S, Sabzian K, et al. Characterization and identification of candiduria due to Candida species in diabetic patients. Curr Med Mycol 2016;2:10-4.  Back to cited text no. 32
    
33.
Zomorodian K, Kavoosi F, Pishdad G, Mehriar P, Ebrahimi H, et al. Prevalence of oral Candida colonization in patients with diabetes mellitus. J Mycol Med 2016;26:103-10.  Back to cited text no. 33
    
34.
Khedri S, Santos A, Roudbary M, Hadighi R, Falahati M, et al. Iranian HIV/AIDS patients with oropharyngeal candidiasis: Identification, prevalence and antifungal susceptibility of Candida species. Lett Appl Microbiol 2018;67:392-9.  Back to cited text no. 34
    
35.
Roudbary M, Roudbarmohammadi S, Bakhshi B, Farhadi Z, Nikoomanesh F. Identification of Candida species isolated from Iranian women with vaginal candidiasis by PCR-RFLP method. Eur J Exp Biol 2013;3:365-9.  Back to cited text no. 35
    
36.
Bii C, Kangogo M, Revathi G, Wanyoike M. Genotypes of Candida albicans from clinical sources in Nairobi Kenya. Afr J Microbiol Res 2009;3:475-7.  Back to cited text no. 36
    
37.
Tang HJ, Liu WL, Lin HL, Lai CC. Epidemiology and prognostic factors of candidemia in elderly patients. Geriatr Gerontol Int 2015;15:688-93.  Back to cited text no. 37
    
38.
Hammad MM, Darwazeh AM, Idrees MM. The effect of glycemic control on Candida colonization of the tongue and the subgingival plaque in patients with Type II diabetes and periodontitis. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2013;116:321-6.  Back to cited text no. 38
    
39.
Rajakumari ML, Kumari PS. Prevalence of Candida species in the buccal cavity of diabetic and non-diabetic individuals in and around Pondicherry. J Mycol Med 2016;26:359-67.  Back to cited text no. 39
    
40.
Silva S, Henriques M, Hayes A, Oliveira R, Azeredo J, et al. Candida glabrata and Candida albicans co-infection of an in vitro oral epithelium. J Oral Maxillofac Pathol 2011;40:421-7.  Back to cited text no. 40
    
41.
Rauceo JM, Gaur NK, Lee KG, Edwards JE, Klotz SA, et al. Global cell surface conformational shift mediated by a Candida albicans adhesin. Infect Immun 2004;72:4948-55.  Back to cited text no. 41
    
42.
Zakikhany K, Naglik JR, Schmidt-Westhausen A, Holland G, Schaller M, et al. In vivo transcript profiling of identifies a gene essential for interepithelial dissemination. Cell Microbiol 2007;9:2938-54.  Back to cited text no. 42
    
43.
Silva-Dias A, Miranda IM, Branco J, Monteiro-Soares M, Pina-Vaz C, et al. Adhesion, biofilm formation, cell surface hydrophobicity, and antifungal planktonic susceptibility: Relationship among Candida spp. Front Microbiol 2015;6:205.  Back to cited text no. 43
    
44.
Nett JE, Lepak AJ, Marchillo K, Andes DR. Time course global gene expression analysis of an in vivo Candida biofilm. J Infect Dis 2009;200:307-13.  Back to cited text no. 44
    
45.
Pierce CG, Vila T, Romo JA, Montelongo-Jauregui D, Wall G, et al. The Candida albicansbiofilm matrix: Composition, structure and function. J Fungi 2017;3:14.  Back to cited text no. 45
    
46.
Soysa NS, Samaranayake L, Ellepola A. Diabetes mellitus as a contributory factor in oral candidosis. Diabet Med 2006;23:455-9.  Back to cited text no. 46
    
47.
Tsang C, Chu F, Leung W, Jin L, Samaranayake L, et al. Phospholipase, proteinase and haemolytic activities of Candida albicans isolated from oral cavities of patients with Type 2 diabetes mellitus. J Med Microbiol 2007;56:1393-8.  Back to cited text no. 47
    
48.
Silva S, Rodrigues CF, Araújo D, Rodrigues ME, Henriques M. Candida species biofilms' antifungal resistance. J Fungi 2017;3:8.  Back to cited text no. 48
    
49.
Rossoni RD, Barbosa JO, Vilela SF, Jorge AO, Junqueira JC. Comparison of the hemolytic activity between C. albicans and non-albicans Candida species. Braz Oral Res 2013;27:484-9.  Back to cited text no. 49
    
50.
Manfredi M, McCullough M, Al-Karaawi ZM, Hurel S, Porter S. The isolation, identification and molecular analysis of Candida spp. isolated from the oral cavities of patients with diabetes mellitus. Oral Microbiol Immunol 2002;17:181-5.  Back to cited text no. 50
    
51.
Rajendran R, Robertson DP, Hodge PJ, Lappin DF, Ramage G. Hydrolytic enzyme production is associated with Candida albicans biofilm formation from patients with Type 1 diabetes. Mycopathologia 2010;170:229-35.  Back to cited text no. 51
    
52.
Luo G, Samaranayake LP, Yau JY. Candida species exhibit differential in vitro hemolytic activities. J Clin Microbiol 2001;39:2971-4.  Back to cited text no. 52
    
53.
de Paula Menezes R, Ferreira JC, de Sá WM, de Aquino Moreira T, Malvino LD, de Araujo LB, et al. Frequency of Candida species in a tertiary care hospital in Triangulo Mineiro, Minas Gerais State, Brazil. Rev Inst Med Trop Sao Paulo 2015;57:185-91.  Back to cited text no. 53
    
54.
Imran ZK, Al-Asadi YF. Comparison of 25S and 18S primers for estimating clinical isolates of Candida diversity and evaluate their virulence in eyes infections in Iraq. Appl Sci Report 2015;9:166-71.  Back to cited text no. 54
    
55.
Gong YB, Jin B, Qi H, Zhang R, Zhang XY, et al. Multilocus sequence typing of Candida albicans isolates from the oral cavities of patients undergoing haemodialysis. Sci Rep 2018;8:1-8.  Back to cited text no. 55
    
56.
Kumar M, Mishra NK, Shukla PK. Sensitive and rapid polymerase chain reaction based diagnosis of mycotic keratitis through single stranded conformation polymorphism. Am J Ophthalmol 2005;140:851-7.e852.  Back to cited text no. 56
    
57.
Zhao X, Oh SH, Jajko R, Diekema DJ, Pfaller MA, et al. Analysis of ALS5 and ALS6 allelic variability in a geographically diverse collection of Candida albicans isolates. Fungal Genet Biol 2007;44:1298-309.  Back to cited text no. 57
    


    Figures

  [Figure 1], [Figure 2]
 
 
    Tables

  [Table 1], [Table 2]



 

Top
Previous article  Next article
 
  Search
 
Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
Access Statistics
Email Alert *
Add to My List *
* Registration required (free)

 
  In this article
Abstract
Introduction
Materials and Me...
Results
Discussion
References
Article Figures
Article Tables

 Article Access Statistics
    Viewed146    
    Printed0    
    Emailed0    
    PDF Downloaded25    
    Comments [Add]    

Recommend this journal