Natural defenses against Candida colonization breakdown in the oral cavities of the elderly.

Candida colonization of the oral cavity increases in the elderly. A major predisposing condition is denture use, which also increases in the elderly. To test whether the increase in colonization is age-related in a fashion independent of denture use, we analyzed the frequency (incidence) of carriage, the intensity of carriage, the multiplicity of species, and the genetic relatedness of strains in the oral cavities of 93 test subjects separated into the three age groups: 60 to 69 yr, 70 to 79 yr, and > or = 80 yr. Each age group was further subdivided into subjects with and without dentures, and into males and females. The results demonstrate that the frequency of carriage, the intensity of carriage, and multispecies carriage all increase as a function of age and differ according to gender, in both cases independent of denture use, suggesting that the natural suppression of yeast carriage in the oral cavity breaks down in the elderly. In addition, it is demonstrated that Candida glabrata colonizes the oral cavities of elderly individuals without dentures only after 80 yr of age, suggesting that there are age-related compromising conditions other than denture use in this most elderly age group.     

Extracellular proteinases of Candida species pathogenic yeasts

The increased incidence of severe disseminated infections caused by the opportunistic yeast‐like fungi Candida spp. highlights the urgent need for research into the major virulence factors of these pathogens—extracellular aspartic proteinases of the candidapepsin and yapsin families. Classically, these enzymes were considered to be generally destructive factors that damage host tissues and provide nutrients for pathogen propagation. However, in recent decades, novel and more specific functions have been suggested for extracellular candidal proteinases. These include contributions to cell wall maintenance and remodeling, the formation of polymicrobial biofilms, adhesion to external protective barriers of the host, the deregulation of host proteolytic cascades (such as the complement system, blood coagulation and the kallikrein–kinin system), a dysregulated host proteinase–inhibitor balance, the inactivation of host antimicrobial peptides, evasion of immune responses and the induction of inflammatory mediator release from host cells. Only a few of these activities recognized in Candida albicans candidapepsins have been also confirmed in other Candida species, and characterization of Candida glabrata yapsins remains limited.     

Prevalence of Candida albicans in oral cavities and root canals of children

The aim of this study was to examine the prevalence of C. albicans in the oral cavities and root canals of children. Twenty healthy and caries-free children and 13 children with caries, were screened. Imprint samples and sterile paper points were used to obtain the samples from oral cavities and root canals respectively. The production of germ tubes and the development of chlamydospores identified yeast cultures. Sixty-nine percent of children with caries and 5% of caries-free children were found to be Candida carriers. The difference in candidal prevalence between two groups was significant (p < 0.05). Sixty-one point five percent of children were positive for Candida in the root canal. Since, increase in the C. albicans in the oral cavity provides a potential source of the fungus particularly when resistance falls below a certain threshold, attention to strategies for the reduction of this pervasive and persistent pathogen becomes important. Therefore, reduction of caries and or introduction of antifungal agents during root canal treatment of children may be offered.     

Implantation of Candida albicans and other Candida species in the oral cavity of rats

The carriage of five Candida species in the mouths of normal and siaioadenectomised rats was determined for periods up to 30 days after inoculation into the oral cavity. In both test and control animals. Candida albicans was the species recovered in greatest quantities at all periods, followed by C. parapsilosis and C. tropicalis. In contrast. C. gullliermondii and C. krusei were isolatable only in small numbers and only from the 1st up to the 5th day; they were not present thereafter. Sialoadenectomy favoured oral colonisation only by C. albicans ( P <0.05) and did not influence the carriage of the other species.     

Bacteria and Candida yeasts in inflammations of the oral mucosa in children with secondary immunodeficiency

J Oral Pathol Med (2012) 41 : 568–576 Background: Oral microbial flora and a damaged oral mucosa may increase the risk of bacteriemia, fungemia and complications in immunocompromised patients. Aim of the Study: Assessment of presence: bacteria and Candida spp. in different oral lesions, and the incidence of bacteremia in the case of a damaged mucosa in transplant recipients and patients receiving anti‐tumour chemotherapy. Material and Method: Forty‐five patients – 18 months to 18 years of life, were included (20 – organ recipients, 14– anti‐tumour chemotherapy, 11 – control group). Clinical, oral mucosa examination focused on the type, severity and site of lesions, and microbiology assessed the presence of bacteria and fungi in the material from lesions. Blood cultures were performed in ten immunocompromised patients with manifestations of systemic infection. The control material consisted of blood cultures made prior to the onset of oral lesions and after 4–6 weeks following their remission in a diagnosed bacteremia. The statistical analysis was performed. Results: In the subjects with secondary immunodeficiency, among other coagulase‐negative Staphylococcus (CoNS), Candidia spp. were more frequent. In cancer patients, mucositis was associated with Candida spp., Streptococcus spp. Organ recipients with stomatitis exhibited the presence of CoNS, Streptococcus viridians and other. Oral lesions in the control group contained Haemophilus parainfluenzae, Neisseria spp. and Staphylococcus aureus. In 30% of immunocompromised patients, oral lesions were accompanied by bacteremia. Conclusions: A correlation has been found between oral lesions and the presence of S. aureus in patients without secondary immunodeficiency, and of CoNS, Enterococcus spp., Candida spp. in immunocompromised patients.     

Coaggregation of Candida albicans oral Actinomyces species

Eight strains of Actinomyces were examined for their ability to coaggregate in vitro with four strains of Candida albicans. The Actinomyces coaggregated to various degrees with all of the Candida strains. Exposure of the Candida but not the Actinomyces to heat, trypsin, proteinase K, amphotericin B or trichodermin abolished Coaggregation. All sugars tested did not inhibit any of the reactions. All coaggregating pairs were disaggregated by the addition of SDS, but nonionic detergents had no effect. The addition of urea or EDTA completely reversed Coaggregation. Actinomyces strains were sensitive to periodate oxidation, whereas the Candida strains were unaffected. These data suggest that the coaggregations involve a protein on the Candida surface that may interact with carbohydrates or carbohydrate-containing molecules on the surface of the Actinomyces. These observations expand the known range of intergeneric coaggregations occurring between human oral microbes and indicate that Coaggregation of C. albicans and Actinomyces may be an important factor in oral colonization by this yeast.     

In vitro secreted aspartyl proteinase activity of Candida albicans isolated from oral diseases and healthy oral cavities

The in vitro secreted aspartyl proteinase (SAP) activity of Candida albicans isolated from a variety of oral conditions, including healthy oral cavities, was determined. SAP activity (units/10(6) cells/ml, +/-SD) was 0.28 +/- 0.33 for pseudomembranous candidosis isolates (n = 18), 0.35 +/- 0.46 for chronic erythematous candidosis isolates (n = 21) and 0.30 +/- 0.32 for chronic hyperplastic candidosis isolates (n = 50). SAP activity of 0.19 +/- 0.22 was recorded for isolates from squamous cell carcinoma (n = 18), 0.26 +/- 0.37 for burning mouth syndrome isolates (n = 29), 0.25 +/- 0.38 for isolates from xerostomia (n = 15) and 0.39 +/- 0.50 for isolates from lichen planus (n = 13). The SAP activity of isolates from oral disease states was significantly (P < 0.05) higher than that recorded for 28 isolates from healthy mouths (activity of 0.04 +/- 0.03). However, there was no significant difference in the SAP activity between the three forms of clinical oral candidosis (P > 0.05). SAP activity was inhibited in control samples containing the SAP inhibitor, pepstatin A. These results indicate that C. albicans strains associated with oral disease have inherently higher SAP activity.     

Betel quid-associated oral lesions and oral Candida species in a female Cambodian cohort

BACKGROUND: Betel quid chewing (BQC) is still prevalent among elderly Cambodian women and is associated with a wide variety of oral mucosal lesions. BQC has also been associated with a reduced rate of dental caries and changes in the oral microbiological flora. METHODS: Since no studies were available on the impact of BQC on the oral carriage of Candida species, in this study oral swabs (Fungiquick, Hain Diagnostika, Germany) were taken from the tongue and palate of 48 Cambodian women with BQC habit (study group) and 13 control subjects without BQC habit (control group) to determine the spectrum of Candida species in these two groups. In addition, we investigated lesions of the oral mucosa likely to be associated with BQC habit in both study and control groups. RESULTS: The median duration of BQC was 10 years (range 10 months-30 years). The following oral lesions were found in the study group: betel chewer's mucosa (85.4%), oral leukoplakia (8.3%), leukoedema (37.5%) and oral lichen planus (4.2%). Oral candidiasis was seen neither in BQ-chewers nor in controls. Candida spp. were found in 70.8% of the cases (controls 69.2%). Whilst C. albicans was isolated from 27.1% of the study cohort, C. tropicalis was the second most common isolate. One control case was colonised by C. dubliniensis--the first report of this organism from a Cambodian population. There was no significant difference in the candidal carriage rate or the Candida species isolated between the study and the control group. CONCLUSIONS: Mycological findings from the present study do not indicate that BQC has a significant effect on oral colonisation by Candida species.     

Fluconazole-resistant Candida species in the oral flora of fluconazole-exposed HIV-positive patients

The purpose of this study was to examine the effect of preceding fluconazole treatment on the oral mycologic flora and on the sensitivity of oral Candida albicans isolates to fluconazole. Saline oral rinses were collected from 89 HIV-positive patients, of whom 48 had been exposed to fluconazole and 41 were fluconazole-naive. The rinses were cultured on Sabouraud's and Pagano Levin agars, and yeasts were identified by standard methods. Fluconazole sensitivity of C. albicans isolates was measured by disk diffusion assay. C. albicans was isolated from 69% of patients who had received fluconazole and from 93% of the patients who were fluconazole-naive (p < 0.05). Nine other species of yeasts were also isolated, most commonly C. glabrata. Five patients previously exposed to fluconazole harbored fluconazole-resistant C. albicans, whereas no resistance was detected among the patients who were fluconazole-naive (p < 0.01). Sixteen of the patients who were fluconazole-exposed carried yeasts other than C. albicans, compared with only five patients in the fluconazole-naive group (p < 0.01). All of the fluconazole-resistant strains were isolated from patients with low CD4 counts (less than 100 cells/ml) and after lengthy fluconazole exposures. Nevertheless, patients in Charlotte, N.C., who had a greater mean fluconazole exposure time (10.25 ± 1.41 months) than patients in Glasgow, UK, (0.65 ± 0.18 months; p < 0.005), did not develop significantly more in vitro resistance or species diversity. This study indicates that long-term fluconazole treatment can have significant effects on the yeast flora of the mouth, particularly in a patient with a CD4 count of less than 100 cells/ml.     

Susceptibility of Candida albicans isolates from the oral cavities of HIV-positive patients to histatin-5

STATEMENT OF PROBLEM: Oral surfaces, including the denture-fitting surface, may serve as a reservoir for disseminated candidal infections, particularly in immunocompromised hosts such as patients with AIDS. Histatins are a group of small, cationic antifungal peptides present in human saliva. There is limited information on the antifungal activity of peptides against Candida albicans isolates from HIV-positive patients. PURPOSE: This study investigated the fungicidal effects of histatin-5 against oral isolates of C. albicans from HIV-positive and HIV-negative patients. MATERIAL AND METHODS: An isolate of C. albicans from each of 2 HIV-positive patients (both male) and 3 HIV-negative patients (2 male and 1 female) was obtained. American Type Culture Collection 90028 served as a reference strain. All isolates were identified with sugar assimilation tests and the germ tube test. Fungicidal assays were performed on exponential C. albicans cells in the presence or absence of 0.315 to 50 microm of histatin-5. Numerical data were subjected to 1-way analysis of variance and Tukey's multiple range test (P<.05). RESULTS: Histatin-5 (50 microm) killed more than 95% of C. albicans isolates from HIV-negative patients and more than 90% of isolates from the reference strain. The same treatment induced 75.3% and 66.1% loss of viability in C. albicans isolates taken from HIV-positive patients (A1 and A2 cells, respectively). The difference between the fungicidal effects in the HIV-positive and HIV-negative groups was significant. (P<.05). CONCLUSION: Within the limited population of this study, C. albicans isolates from the oral cavities of HIV-positive patients were less sensitive to histatin-5 than oral isolates from HIV-negative patients.     

Close association between oral Candida species and oral mucosal disorders in patients with xerostomia

Oral Diseases (2012) 18, 667-672 Objective: Heightened interest in oral health has lead to an increase in patients complaining of xerostomia, which is associated with various oral mucosal disorders. In this study, we investigated the relationship between Candida species and oral mucosal disorders in patients with xerostomia. Subjects and Methods: We evaluated whole salivary flow rate and presence of oral mucosal disorders in 48 patients with xerostomia and 15 healthy controls. The number of Candida species was measured as colony-forming units after propagation on selective medium. Identification of Candida at the species level was carried out by polymerase chain reaction and restriction fragment length polymorphism analysis. We then examined the relationship between Candida species and oral mucosal symptoms. Results: Compared with controls, patients with xerostomia exhibited significantly decreased whole salivary flow rate, increased rate of oral mucosal symptoms, and higher numbers of Candida. Salivary flow rate negatively correlated with the number Candida. Among patients with oral candidiasis, Candida albicanswas isolated from the tongue mucosa and Candida glabratawas isolated from the angle of the mouth. Conclusion: These results suggest that particular Candida species are involved in the pathogenesis of oral mucosal disorders in patients with xerostomia.     

Improved oral hygiene and Candida species colonization level in geriatric patients

OBJECTIVES: This work consists in improving oral hygiene (OH) for elderly dependent people in long-term hospital care, in order to decrease the degree of colonization and the associated risk of developing oral candidiasis. As this population frequently suffers from such colonization and because it is difficult to install and practice OH care, a study protocol was designed at the request of geriatricians. The objective of the present study was to set up a programme of OH, applied by the care staff, and to monitor oral colonization of by Candida spp. BASIC RESEARCH DESIGN: We compared the levels of hygiene and Candida spp. colonization for a group of 110 long-term patients in geriatric departments at T1, when clinical data were collected and oral mycological samples taken before the OH protocol was applied, and at T2, during the postprotocol phase after 3 months of application, when the clinical data and sample collection were repeated. RESULTS: During these 3 months 11 patients died. These patients were excluded from the results, which are presented for matched series of the 99 patients still present at T2. Statistical analysis comparing the clinical and biological parameters at T1 and T2 established that there had been an improvement in OH: the 'adequate' level was reached for 72.4% of patients at T2 compared with 41.8% at T1 (P < 0.001) and the 'very inadequate' level was observed for 9.2% at T2 compared with 27.9% at T1 (P < 0.01). A reduction was observed in the number of patients showing the highest degree of C. albicans and C. glabrata colonization (> 50 colony forming units) from 41.9% at T1 to 24.9% at T2 (P < 0.05) and from 56.4% at T1 to 13.0% at T2 (P < 0.05) respectively. The number of patients with candidiasis fell significantly from 43.2% at T1 to 10.2% at T2. CONCLUSIONS: The OH protocol led to an overall decrease in Candida spp. colonization, a significant reduction in the number of candidiasis and an improvement in the level of oral and denture hygiene but vigilance is still necessary concerning OH care and the initial training of staff in specific care of the mouth.     

Biofilm formation by oral clinical isolates of Candida species

We have conducted a longitudinal study to quantify biofilms in oral clinical isolates of Candida species (spp.) from adults with local and systemic predisposing factors for candidiasis. A total of 69 yeast isolates from 63 Mexican patients were evaluated. These isolates (39 C. albicans, 15 C. tropicalis, 7 C. glabrata, 4 C. krusei, 1 C. lusitaniae, 1 C. kefyr, 1 C. guilliermondii and 1 C. pulcherrima) were obtained from two clinical sites: 62.3% (n = 43) from the oral mucosa of totally and partially edentulous patients, and 37.7% (n = 26) from the oral mucosa of diabetics. In addition, Candida ATCC strains were used as controls for each experiment. The kinetics of biofilm formation were measured by 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide [XTT] reduction; each isolate was tested at 6, 12 and 24 h. Biofilm formation is dependent on the Candida spp. and its clinical origin. On average, the oral isolates of C. glabrata are strong biofilm producers, whereas C. albicans and C. tropicalis are moderate producers. The most common species in our population was C. albicans. While the kinetics of C. albicans biofilm formation varies between oral isolates, it generally maintains steady growth from 2 to 48 h, when it reaches its maximum growth.     

Prevalence of oral Candida species in leprosy patients from Cambodia and Thailand

BACKGROUND: Leprosy is a chronic bacterial infection which may lead to significant orofacial morbidity. However, reports on the oral mycotic flora of leprosy patients are rare. The aim of the current study was to explore the oral yeast carriage in two groups of leprosy patients. METHODS: 40 Cambodian (seven men, 33 women) and 48 Thai (14 men, 34 women) leprosy patients from Leprosy Rehabilitation Centre Khien Kleang, Phnom Penh, Cambodia and McKean Rehabilitation Center, Chiangmai, Thailand were randomly selected and their demographic data and clinical history were recorded. Tongue and palatal swabs of each patient were collected using sterile Fungi-Quick swabs (Hain Diagnostika, Nehren, Germany) and they were cultured aerobically on Sabouraud's dextrose agar and CHROMAgar (CHROMagar, Paris, France). Yeast were identified by germ tube, chlamydospore production, and assimilation tests (API 20C AUX, Bio-Merieux, Marcy l'Etoile, France) and reconfirmed using APILAB Plus system (Bio-Merieux). RESULTS: Two groups (Cambodian and Thai) had median age of 35 and 64 years. They had been with leprosy for median durations of 17.7 and 38.9 years (P<0.05), respectively. Overall yeast carriage in two cohorts were 80% and 93.75%. Candida albicans had highest carriage rate in either group (65.6%, 44.4%). Candida krusei and C. glabrata existed as second-line colonizers after C. albicans. Candida glabrata carriage was significantly higher in Thai patients (P<0.05). Multispecies carriage was seen in three Cambodian (9.4%) and five Thai (11.5%) patients. CONCLUSIONS: This study indicates high oral yeast carriage in leprosy patients. Candida albicans remains predominant while C. krusei and C. glabrata are second-line oral colonizers. Co-inhabitation of multiple yeast species is also noted in these patients' oral mycotic flora.     

Susceptibility of oral Candida species to calcium hydroxide in vitro.

AIM: The susceptibility of common oral Candida species to saturated aqueous calcium hydroxide solution was studied. METHODOLOGY: The yeast species tested were C. albicans (16 strains). C. glabrata (three strains), C. guilliermondii (three strains), C. krusei (two strains), and C. tropicalis (two strains). At least one reference strain of each species was used; the others were clinical isolates either from persistent apical periodontitis or from marginal periodontitis. The susceptibility of Enterococcus faecalis (ATCC 29212) was studied for comparative purposes. Standardized inocula of the strains were incubated in aqueous calcium hydroxide solution, pH 12.4, for time-periods ranging from 5 min to 6 h. Volumes of 0.1 mL of the test suspension were cultured directly on Brucella blood agar and incubated in air at 37C. The plates were inspected for growth at 24 and 48 h and the colonies were counted. The time required to reduce the number of colony-forming units to less than 0.1% of the initial number was determined for each strain. RESULTS: The sensitivity of the C. albicans strains was generally low, with 16 h of incubation required to kill 99.9% of the colony-forming units. No differences in susceptibility between C. albicans strains isolated from root-canal infections and from periodontitis were found. Both strains of C. tropicalis were killed between 3 and 6 h of incubation, whilst strains of C. guilliermondii were killed after only 1020 min of incubation. All strains of C. glabrata survived 20 min, but not 1 h, of incubation, whilst 13 h were required to kill C. krusei. Compared with E. faecalis, all Candida spp. showed either equally high or higher resistance to aqueous calcium hydroxide. CONCLUSIONS: This study indicates that Candida spp. are resistant to calcium hydroxide in vitro, which may explain the isolation of yeasts from cases of persistent apical periodontitis.     

Prevalence and antifungal susceptibility profiles of Candida glabrata,Candida parapsilosis and their close-related species in oral candidiasis

ObjectiveTo evaluate the importance of Candida glabrata, Candida parapsilosis and their close-related species, Candida bracarensis, Candida nivariensis, Candida metapsilosis and Candida orthopsilosis in patients with oral candidiasis and, to determine the in vitro activities of antifungal drugs currently used for the treatment.MethodsOne hundred fourteen isolates of C. glabrata and 97 of C. parapsilosis, previously identified by conventional mycological methods, were analysed by molecular techniques. In vitro antifungal susceptibility to fluconazole, itraconazole, miconazole, and nystatin was evaluated by CLSI M44-A2 disk diffusion test, and by CLSI M27-A3 microdilution for fluconazole.ResultsAll C. glabrata isolates were identified as C. glabrata sensu stricto, 93 out of 97 C. parapsilosis isolates as C. parapsilosis sensu stricto, three as C. orthopsilosis and one as C. metapsilosis. Candida glabrata was mainly isolated in mixed cultures but C. parapsilosis complex was more frequent in pure culture. Candida metapsilosis and C. orthopsilosis were isolated as pure culture and both species were susceptible to all antifungal agents tested. Most C. glabrata isolates were susceptible to miconazole and nystatin, but resistant to fluconazole and itraconazole. Azole cross resistance was also observed. Candida parapsilosis isolates were susceptible to fluconazole although azole cross resistance to miconazole and itraconazole was observed.ConclusionThis study highlights the importance of accurate identification and antifungal susceptibility testing of oral Candida isolates in order to have an in-depth understanding of the role of C. glabrata and C. parapsilosis in oral candidiasis.     

The relationship between oral hygiene and oral colonization with Candida species in healthy adult subjects*

To cite this article:
Int J Dent Hygiene
DOI: 10.1111/j.1601‐5037.2009.00371.x
Darwazeh AM‐G, Hammad MM, Al‐Jamaei AA. The relationship between oral hygiene and oral colonization with Candida species in healthy adult subjects. Abstract: Poor oral hygiene has been frequently suggested as a predisposing factor for oral candidal colonization, but the convincing evidence is lacking. Objective: To assess and compare oral candidal colonization, both quantitatively and qualitatively, in groups of healthy dentate subjects with different levels of oral hygiene as determined by the plaque index (PI) and gingival index (GI) scores. Methods: The concentrated oral rinse technique was used to isolate Candida species from 149 healthy dentate subjects. Candida species were cultured on Sabouraud’s dextrose agar plates and identified by germ‐tube test and the automated Vitek® system biochemical yeast card. According to the PI and GI scores, subjects were divided into different groups of oral hygiene level. Results: Candida species were isolated from 86 (57.7%) subjects. The prevalence of candidal carriage increased significantly as a function of age (P = 0.023), but was comparable between males and females (58.7% and 56.7% respectively; P = 0.87). Oral candidal carriage rate and density were not affected by the levels of dental plaque or gingival condition. The prevalence of oral candididal carriage was significantly higher in the subjects who were not using dental floss compared with those who were using dental floss (P = 0.032). Conclusion: Oral hygiene status, as determined by the PI and the GI scores per se, does not affect oral candidal colonization in healthy dentate subjects.