Elsevier

Archives of Oral Biology

Volume 53, Issue 12, December 2008, Pages 1172-1178
Archives of Oral Biology

Genetic diversity and exoenzyme activities of Candida albicans and Candida dubliniensis isolated from the oral cavity of Brazilian periodontal patients

https://doi.org/10.1016/j.archoralbio.2008.06.003Get rights and content

Abstract

Objective

Mucosal surfaces are the primary oral reservoirs of Candida species, but these species can also be found in subgingival biofilm. The present study investigated the genetic diversity and production of exoenzymes of C. albicans and C. dubliniensis isolated from the oral cavity of systemically healthy patients with periodontitis.

Design

Fifty-three patients were analysed. Samples were collected from three oral cavity sites (periodontal pocket, gingival sulci and oral mucosa), plated and, after isolation, suspect strains of C. albicans and C. dubliniensis were identified by PCR. The genetic diversity of the isolates was evaluated by RAPD and the activities of the secreted aspartyl proteinases and phospholipases were evaluated by the agar plate method.

Results

Twenty-one patients showed positive results for Candida spp. There were no statistically significant differences between genders, or between sites. C. albicans was the most frequently found specie, while C. dubliniensis was isolated from the periodontal pocket of only one patient. Sixteen genotypes were detected among the C. albicans isolates, and one among the C. dubliniensis isolates. The similarity coefficient (SSM) values among the C. albicans genotypes ranged from 0.684 to 1.0 with an average of 0.905 ± 0.074. All isolates produced high levels of Saps and most of them produced high levels of phospholipases. No relationship was found between the genotypes and the pattern of enzymatic production. There was no association between specific genotypes and their site of isolation.

Conclusions

The results of the present study suggest that genetically homogeneous strains of C. albicans are present in the oral cavity of patients with periodontitis and that these strains are capable of producing high levels of exoenzyme.

Introduction

Candida species are opportunistic pathogens that live as benign members of the commensal microbiota in the oral cavity of a significant proportion of healthy individuals.1 Mucosal surfaces are the primary oral body reservoirs of these yeasts, but they can also be found in dental plaque.2 Such yeasts become pathogenic in response to physiological changes in the host, particularly immunocompromised individuals, resulting in oral candidosis or invasive systemic infection.3

Candida albicans has been found in the subgingival biofilm of periodontal pockets from different forms of periodontitis,2, 4, 5, 6, 7 from peri-implantitis lesions8 and, especially, from subgingival sites of HIV-positive patients.9, 10, 11C. albicans presents a range of virulent properties that are potentially relevant to periodontal infections, such as the capacity to adhere to the epithelium and to invade the gingival conjunctive tissue5, 6, 12, 13 and, particularly, to produce exoenzymes.14, 15Candida dubliniensis possesses characteristics that enable its adaptation to the oral environment, such as the capacity to coaggregate with subgingival bacterial species,16 being found in the oral cavity and in subgingival sites of HIV-positive patients.3 However, the colonisation of the oral cavity of HIV-negative patients has also been reported,17, 18, 19 although it is unclear whether subgingival Candida strains participate in the pathogenesis of periodontitis.2, 20, 21

Many researchers have been increasingly interested in acquiring knowledge about medically important fungi and several reports employing different methods of genotyping have been developed to study the pathogenicity, epidemiology and genetics of C. albicans.10, 22, 23 The RAPD (random amplified polymorphic DNA) method has been used for the genotyping of clinical isolates of C. albicans.20, 24 A recent study identified distinct genotypes of yeasts in patients with periodontitis versus oral healthy subjects.2

However, there is little information about virulence factors of C. albicans and C. dubliniensis isolated from the oral cavity of periodontal patients. It has been suggested25 that enzymatic activity may play an essential role in the capacity that this fungus has to establish itself as a colonizing and/or infectious microorganism. The exoenzymes, secreted aspartyl proteinases (Saps) and phospholipases, degrade immunoglobulins and proteins from the extracellular matrix,26, 27 inhibit the fagocitosis of polymorphonuclear neutrophils28 and induce inflammatory reactions.4, 27 In addition, their extracellular enzymatic secretion may be associated with the genotype and source in C. albicans clinical isolates.25

The present study investigated the relationship between the enzymatic activity of Saps and phospholipases and the genetic diversity of C. albicans and C. dubliniensis, isolated from three sites on the oral cavity of systemically healthy periodontal patients: periodontal pocket, gingival sulci and oral mucosa.

Section snippets

Material and methods

This research was approved by the ethical committee in research of the Piracicaba Dental School, State University of Campinas, SP, Brazil.

Results

Twenty-one (39.6%) of the 53 patients were colonised by C. albicans and/or other Candida species in one or more of the investigated oral sites. Fourteen patients were positive for Candida in site A, eight in site B and 20 in site C. Four of eleven smokers were colonised by yeasts. There were no statistically significant differences in the prevalence of these species between gender (p = 0.5922), between oral sites (p = 0.1287), or between smokers and non-smokers (p = 0.8039). C. albicans was the most

Oral colonisation by Candida spp.

Among the 53 periodontal patients analysed, 21 were colonised by Candida spp., in one or more oral sites, and C. albicans was the most frequently isolated species. Such results were consistent with those previously described in the literature.1, 4, 6, 22C. albicans has been isolated from subgingival biofilm in immunocompetent individuals, especially in cases considered as “refractory” to periodontal therapy, such as superinfectant organisms.5, 12 We isolated these species from periodontal

Acknowledgments

This research was supported by FAEP, State University of Campinas, State of São Paulo, Brazil.

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