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L P Samaranayake

Publications and source records attributed to L P Samaranayake.

At least 19 recordsLinked to original sources

The in vitro post-antifungal effect of nystatin on Candida species of oral origin.

The post-antifungal effect (PAFE) is defined as the suppression of growth that persists following limited exposure of yeasts to antimycotics and subsequent removal of the drug. Although limited data are available on the PAFE of nystatin on oral isolates of C albicans, there is no information on non-albicans Candida species. As nystatin is the commonest antifungal agent prescribed in dentistry, the main aim of this investigation was to measure the PAFE of oral isolates of Candida belonging to six different species (five isolates each of C. albicans, C. tropicalis, C. krusei, C. parapsilosis, C. glabrata and C. guilliermondii) following limited exposure (1 h) to nystatin. The yeasts were examined for the presence of the PAFE after 1 h exposure to the minimum inhibitory concentration (MIC) of nystatin. The PAFE was determined as the difference in time (h) required for the growth of the drug-free control and the drug-exposed test cultures to increase to the 0.05 absorbance level following removal of the antifungal agent. The mean duration of nystatin-elicited PAFE was lowest for C. albicans (6.85 h) and greatest for C. parapsilosis (15.17 h), while C. krusei (11.58 h), C. tropicalis (12.73 h), C. glabrata (8.51 h), and C. guilliermondii (8.68 h) elicited intermediate values. These findings clarify another intriguing possibility for the persistent, chronic recurrence of oral C. albicans infections despite apparently adequate antifungal drug regimens. The significant variations in nystatin-induced PAFE amongst non-albicans species may also have clinical implications, in terms of nystatin regimens used in the management of these fungal infections.

Analysis of Variance

Oral manifestations of HIV infection in a group of predominantly ethnic Chinese.

A total of 32 HIV-infected, predominantly ethnic Chinese individuals from Hong Kong were examined for oral mucosal lesions over a period of 1 year. The commonest oral lesion found was minor aphthous ulceration (27.4%), while xerostomia (17.8%), ulceration NOS (not otherwise specified; 12.3%), hairy leukoplakia (11.0%) and erythematous candidiasis (6.9%) were less frequent; Kaposi's sarcoma was notable for its absence. When the relationship between the number of oral lesions with age, risk group, medication taken, CDC staging and CD4+ count of the study group was investigated, a significantly higher number of oral lesions was associated with use of AZT, homosexuals and CDC stage IV; in contrast, a smaller number of lesions was found in those on antiparasitics and multivitamins (all P<0.05). When compared with studies from other parts of the world, the frequency of oral lesions appeared to be less common in the current study group. However, due to a lack of similar studies from the Asian region, especially in ethnic Chinese, it is not clear whether this difference could be attributed to racial, social or geographic factors.

Adult

The effect of limited exposure to antimycotics on the relative cell-surface hydrophobicity and the adhesion of oral Candida albicans to buccal epithelial cells.

Candida albicans is the major aetiological agent of oral candidosis. Adhesion to oral mucosal surfaces is considered a prerequisite for its successful colonization and subsequent infection, and its relative cell-surface hydrophobicity (CSH) is a contributory physical force. Thus, the main aim here was to determine the CSH of 10 isolates of oral C. albicans after a short exposure to sublethal concentrations of four antifungal agents and to correlate these findings with their adhesion to human buccal epithelial cells (BEC). The yeasts were exposed to sublethal concentrations of nystatin [x 6 minimal inhibitory concentration (MIC)], 5-fluorocytosine (x 8 MIC), ketoconazole (x 4 MIC) and fluconazole (x 4 MIC) for 1 h. The drug was then removed, and the CSH and BEC adhesion assessed by a biphasic aqueous-hydrocarbon assay and a microscopic method, respectively. The mean percentage reductions of CSH after exposure to nystatin, 5-fluorocytosine, ketoconazole and fluconazole were 27.14% (p = 0.01), 9.46% (p = 0.43), 19.47% (p = 0.04) and 6.16% (p = 0.59). Similarly, exposure to all the drugs except 5-fluorocytosine resulted in a significant inhibition of yeast adhesion to BEC, with nystatin eliciting the highest and fluconazole the least inhibition. Further, on regression analysis a strong positive correlation was observed between CSH and adhesion to BEC after limited exposure to 5-fluorocytosine (r = 0.48, p < 0.0001), ketoconazole (r = 0.48, p < 0.0001), fluconazole (r = 0.55, p < 0.0001) as well as in the unexposed controls (r = 0.41, p = 0.001), although nystatin was an exception (r = 0.09, p = 0.44). Taken together, these data elucidate further mechanisms by which antimycotics may operate in vivo to suppress candidal pathogenicity.

Adult

Adhesion of oral Candida albicans isolates to denture acrylic following limited exposure to antifungal agents.

Candidal adherence to denture acrylic surfaces is implicated as the first step in the pathogenesis of Candida-associated denture stomatitis, the most prevalent form of oral candidosis in the West. This condition is treated by topically administered antifungal agents, mainly belonging to the polyenes and azoles. As the intraoral concentrations of antifungals fluctuate considerably due to the dynamics of the oral environment, the effect of short exposure to sublethal concentrations of antifungals on the adhesion of Candida albicans to denture acrylic surfaces was investigated. Seven oral C. albicans isolates were exposed to four-eight times minimum inhibitory concentrations (MIC) of five antifungal drugs, nystatin, amphotericin B, 5-fluorocytosine, ketoconazole and fluconazole, for 1 h. After removing the drug (by repeated washing) the adhesion of these isolates to acrylic strips was assessed by an in vitro adhesion assay. Exposure to antifungal agents significantly reduced the adherence of all seven C. albicans isolates to denture acrylic. The mean percentage reductions of adhesion after limited exposure to nystatin, amphotericin B, 5-fluorocytosine, ketoconazole and fluconazole were 86.48, 90.85, 66.72, 65.88 and 47.42%, respectively. These findings indicate that subtherapeutic doses of antifungals may modulate oral candidal colonization. Further, these results may have an important bearing on dosage regimens currently employed in treating oral candidosis.

Adhesiveness

The relative pathogenicity of Candida krusei and C. albicans in the rat oral mucosa.

The relative pathogenicity of Candida krusei and C. albicans was investigated by assessing their colonisation and infectivity of the Sprague-Dawley rat oral mucosa. During an initial 21-week period with intermittent oral inoculation, both Candida spp. demonstrated variable surface colonisation of the oral mucosa. After 3 days of oral inoculation, both yeast species were recovered from all animals. During the 21-week period the mean oral load of C. albicans in the control group of rats varied between (26-274) x 10(1) cfu/ml whereas the two test groups of rats carrying C. krusei CK9 and CK13 had a mean load of (2-10) x 10(1) cfu/ml. Although oral colonisation by C. albicans was greater than that of C. krusei, neither species induced candidal infection during this period. Subsequent immunosuppression of the rats by intramuscular cyclophosphamide (40 mg/kg body weight) initiated C. albicans infection of the dorsal tongue (around the conical papillae area) after 4 weeks, in all of three animals while similar lesions due to C. krusei were seen--albeit after 5-7 weeks--in three of eight animals. Characteristic histological changes of mucosal candidosis were discernible on the lingual mucosa of rats infected with both Candida spp. including parakeratosis, absence of a stratum granulosum, thickened rete ridges, micro-abscess formation and polymorph infiltration of the lingual epithelium. Although both species produced fungal hyphae that penetrated the epithelium up to the prickle cell layer, C. albicans hyphae tended to be relatively more profuse. Taken together these results substantiate, for the first time in an animal model, the clinical evidence that C. krusei, once considered an innocuous commensal, is capable of transforming into an invasive pathogen under conditions of immunosuppression.

Animals

Antimicrobial susceptibility of oral isolates of Enterobacter cloacae and Klebsiella pneumoniae from a southern Chinese population.

The antibiotic susceptibilities of 59 Enterobacter cloacae and 39 Klebsiella pneumoniae human oral isolates collected from a southern Chinese population in Hong Kong were investigated for their susceptibility to eight antibiotics: ampicillin, cephalothin, cefuroxime, ceftazidime, ciprofloxacin, gentamicin, tetracycline and trimethoprim/sulfamethoxazole using the E-Test method for direct quantification of minimum inhibitory concentrations. Most strains were sensitive to all antibiotics except ampicillin and cephalothin. Ampicillin resistance was exhibited by 82% of K. pneumoniae and 69% of E. cloacae isolates. Eighty-eight percent of E. cloacae isolates were resistant to cephalothin. Several strains fell within the intermediate category of sensitivity for ampicillin (E. cloacae and K. pneumoniae), cefuroxime (E. cloacae) and tetracycline (K. pneumoniae). Comparison with other Hong Kong data suggests that resistance rates to cefuroxime, ceftazidime, ciprofloxacin, gentamicin, tetracycline and trimethoprim/sulfamethoxazole exhibited by the oral isolates are generally lower than in enterobacters and Klebsiella spp. isolated from urine, skin and soft tissues in Hong Kong populations.

Adult

Subgingival microbiota of shallow periodontal pockets in individuals after head and neck irradiation.

This study aimed at investigating the subgingival plaque microorganisms of shallow pockets (< or = 5 mm) in subjects who previously received irradiation in the head and neck region for treatment of nasopharyngeal carcinoma. Direct microscopy and anaerobic culture were used. Subgingival paper point samples were taken from 6 tooth-sites (one/sextant) per subject for direct microscopy (n = 108). Another set of paper points was taken from the deepest of the previously selected sites (one per subject) with: group A) no bleeding on probing to the sulcus depth (n = 9) and group B) bleeding on probing to the sulcus depth (n = 6) for microscopic and anaerobic culture study. Under the microscope, the microflora was found to be a complex mixture comprising gram-positive and gram-negative cocci, rods and filaments, fusiforms, curved rods and spirochetes. Low level of fungi were observed and mycelia were occasionally detected. There was no significant variation in the plaque bacterial morphotypes observable according to sites of isolation and no significant difference between group A and group B in morphotypes of the different microflora. The predominant cultivable microflora comprised several species of facultative and obligate anaerobic bacteria: Gemella, Peptostreptococcus, Staphylococcus, Stomatococcus, Streptococcus, Actinomyces, Eubacterium, Lactobacillus, Propionibacterium, Neisseria, Veillonella, Bacteroides, Campylobacter, Capnocytophaga, Fusobacterium, Kingella, Porphyromonas and Prevotella species. There was no difference between the two groups except the significantly higher proportion of Kingella dentrificans isolated from group B sites. However, colonization of the gingival sulcus in these individuals by microbes that are normal flora of: skin (Peptostreptococcus prevotii and Propionibacterium granulosum) and gut (Eubacterium aerofaciens, Fusobacterium mortiferum and Fusobacterium varium) was detected. These findings appear to suggest that the major components of the subgingival microflora of shallow sites in previously head- and neck-irradiated individuals are similar to that of gingivitis sites in the normal population although they may contain bacterial or fungal species uncommon in normal subjects.

Adult

The effect of limited exposure to antifungal agents on the germ tube formation of oral Candida albicans.

Candidal adherence has been implicated as the first step in the pathogenesis of oral candidosis, and germ tube formation by Candida albicans has been attributed as a co-factor that promotes adherence. Oral candidosis is treated with polyenes and the azole group of antifungal agents. As the intraoral concentrations of antifungals fluctuate considerably due to the dynamics of the oral cavity, we investigated the effect of short exposure to sub-lethal concentrations of antifungals on the germ tube formation of Candida albicans. After determining the minimum inhibitory concentration (MIC) of the antifungal agents, ten oral isolates of Candida albicans were exposed to sub-lethal concentrations of nystatin (6xMIC), amphotericin B (8xMIC), 5-fluorocytosine (8xMIC), ketoconazole (4xMIC) and fluconazole (4xMIC), for 1 h. Following removal of the antifungal agent and subsequent incubation in a germ tube-inducing medium, the germ tube formation of these isolates was quantified. When compared with the controls, exposure to nystatin and amphotericin B almost completely inhibited germ tube formation of all the isolates (mean percentage reduction of 97.68 and 97.52%, respectively; P<0.0001), while ketoconazole suppressed this activity to a lesser degree (30.84%; P=0.0174). However, 5-fluorocytosine- and fluconazole-mediated germ tube suppression was minimal (12.63 and 15.93%, respectively; P=0.3255 and P=0.3791). In clinical terms, these findings indicate that short exposure to sub-therapeutic levels of commonly prescribed antifungals may modulate candidal germ tube formation, and thereby the clearance of the organisms from the oral cavity.

AIDS-Related Opportunistic Infections

Adhesion of oral C. albicans to human buccal epithelial cells following limited exposure to antifungal agents.

The major aetiologic agent of oral candidosis is C. albicans, and adhesion to oral mucosal surfaces is considered a vital prerequisite for successful colonisation and subsequent infection by this agent. Although many antimycotics are available for the treatment of oral candidosis, the diluent effect of saliva and the cleansing action of the oral musculature often tend to reduce the availability of the agents below that of the effective therapeutic concentration. Therefore, the yeasts undergo only a limited exposure to the antifungals during therapy. Hence, the main aim of the present study was to determine the in vitro adhesion of ten isolates of oral C. albicans to buccal epithelial cells (BEC) following a short exposure to sublethal concentrations of four antifungal agents. The yeasts were exposed to sublethal concentrations of nystatin (x6 MIC), 5-fluorocytosine (x8 MIC), ketoconazole (x4 MIC) and fluconazole (x4 MIC) for a period of 1 h. Following subsequent removal of the drug, the adhesion of these isolates to BEC was assessed by a previously described adhesion assay. The mean percentage reductions of candidal adhesion to BEC following exposure to sublethal concentrations of nystatin, 5-fluorocytosine, ketoconazole and fluconazole were 72.88%, 16.52%, 40.16% and 24.36%, respectively. Ultrastructural studies revealed that short exposure to nystatin and the azoles (but not 5-fluorocytosine) resulted in aberrant cellular features. These findings indicate that subtherapeutic levels of antifungals may modulate candidal colonisation of the oral mucosa and thereby suppress the invasive potential of the pathogen.

Antifungal Agents

Adhesion to denture acrylic surfaces and relative cell-surface hydrophobicity of Candida parapsilosis and Candida albicans.

C. parapsilosis is an opportunistic emerging pathogen which together with C. albicans causes diseases in immunocompromised patients. Adhesion of Candida species to various surfaces is an important event in colonization and pathogenesis, and the relative cell-surface hydrophobicity (CSH) of the organism is a contributory physical force involved. Therefore, in vitro adhesion to acrylic surfaces and relative CSH of 24 isolates of C. parapsilosis and 10 isolates of C. albicans were studied. There was no significant difference in relative adhesion of C. parapsilosis isolates and C. albicans, although the former demonstrated a tendency for increased adhesion. There was significant intra-species variation in adhesion among isolates of C. parapsilosis (p=0.0001), but not C. albicans. In general, C. parapsilosis isolates demonstrated a two-fold greater relative CSH than C. albicans (p=0.0003). When the relative CSH of superficial and systemic isolates of C. parapsilosis were compared, the former showed a significantly higher (49.15%) relative CSH than their systemic counterparts (p<0.01). A highly significant positive correlation between adhesion and relative CSH of C. parapsilosis (p=0.74, p<0.0001) was also noted. Taken together, these data suggest that the attributes of adhesion and relative CSH of Candida species may contribute differentially in varying disease states of the human host, such as superficial and systemic Candida infections.

Acrylic Resins

Chromosome length polymorphism in clinical isolates of Candida parapsilosis.

Chromosome length polymorphism among 24 clinical isolates of Candida parapsilosis obtained from several human sources was analysed using pulsed-field gel electrophoresis. The isolates, from both superficial and deep infections, comprised a miscellaneous collection from the oral cavity, blood cultures, ear infections, wound secrete, a venous catheter and peritoneal dialysis fluid. Contour-clamped homogenous field electrophoresis using a hexagonal electrode was used for pulsed field gel electrophoresis. The chromosome numbers varied from seven to nine and their sizes ranged from 0.75 to 2.6 Mb. According to the electrophoretic karyotype patterns the 24 isolates could be divided into 9 profiles. However, the majority (18 isolates) fell into 3 groups comprising 7, 8 and 9 chromosomes, containing 5, 11, and 2 isolates, respectively. The remaining six isolates, all of which were from either an oral or another superficial site of isolation, could be categorized into a further six groups. These data confirm previous observations on the genomic heterogeneity of clinical isolates of C. parapsilosis, and illustrate the possible commonality in strains from related clinical habitats.

Candida

Miconazole lacquer in the treatment of denture stomatitis: clinical and microbiological findings in Chinese patients.

OBJECTIVE: To investigate the efficacy of a miconazole lacquer in producing mycological and clinical cure of Candida-associated denture stomatitis in a Chinese cohort and to study the microbiology of the conditions. DESIGN: A cross-sectional study of the clinical and microbiological features of the condition and its response to treatment with the lacquer. SUBJECTS AND METHOD: Twenty-one Chinese adults with palatal erythema; 15 denture wearers matched for age and sex as controls for microbiology. Clinical examination, impression for culture, photograph of palate to monitor response to treatment, and lacquer application on upper denture base on days 0, 7 and 14. Impression and photograph repeated on day 21; impression cultures quantified by image analysis. Palatal impression in controls; dummy impression as control for impression culture procedures. MAIN OUTCOME MEASURES: Severity of lesion, incidence and type of microorganism at presentation and during treatment; qualitative and quantitative assessment of yeast growth; the latter by statistical analysis of pixel units of yeast colonies; correlation of clinical and mycological cure. RESULTS: Out of the 21 lesions (16 Newton's stage I and 5 stage II), 57% had yeasts at presentation, 10% coliforms and 33% no significant growth. Clinical cure in 3 weeks in 71%, mycological cure in 75% in 7 days; clinical and mycological cure coincided only in 43%. Polymicrobial growth of yeasts and coliforms throughout the course in some. CONCLUSIONS: Miconazole lacquer was a viable treatment option for Candida-associated denture stomatitis in this adult, Chinese cohort; microbiology comprised yeasts, mainly C. albicans and coliforms, especially Klebsiella and Enterobacter species; the pathogenic role of the latter is questionable.

Adult

A microbiological study of pre- and postoperative apicoectomy sites.

There is little information on the microbiology of periapical lesions, and no data on the residual microbial flora in the periapex, if any, after apicoectomy procedures. Hence, 64 patients treated by apicoectomy procedures were prospectively studied to assess the bacterial flora in the periapex and to evaluate the residual bacteria in postoperative apicoectomy sites. Of the 64 lesions studied, 14 (22%) were sterile and 50 (78%) yielded bacteria preoperatively. Bacteria could be recovered from 28 (56%) of the latter lesions after apicoectomy and curettage. A total of 105 bacterial strains was isolated from 50 lesions, yielding a range of 1-4 (mean 2.1) species per sample. The isolates comprised 84 (80%) facultative anaerobes and 21 (20%) strict anaerobes. A polymicrobial growth was obtained from 39 lesions whilst 11 lesions yielded pure cultures. On detailed microbiological analyses of 29 lesions, 40% of the isolates were identified as alpha-haemolytic streptococci, half of which were Streptococcus sanguis; anaerobic streptococci were the predominant anaerobes. None of the organisms or group(s) of organisms emerged as recalcitrant colonisers which were difficult to dislodge after surgical debridement. These data indicate that the majority of periapical lesions harbour a variety of flora which cannot be eradicated despite thorough apicoectomy procedures.

Actinomyces

Effects of dietary sugars and, saliva and serum on Candida bioflim formation on acrylic surfaces.

The effect of two dietary sugars, glucose and galactose, on biofilm formation of the oral fungal pathogen Candida on denture acrylic strips coated with saliva and serum pellicles was examined in vitro using Candida albicans (3 isolates), C. glabrata (2 isolates) and C. tropicalis (2 isolates). The degree of biofilm activity was affected by both the dietary sugar and the nature of the pellicle (ANOVA, p < 0.01). With most isolates the glucose grown yeasts demonstrated significantly more bioflim activity than the galactose grown fungi, in the presence of pellicles (ANOVA, p < 0.01 or P < 0.01). In contrast, one isolate of galactose-grown yeast elicited significantly higher biofilm activity than glucose-grown yeasts on the control strips (ANOVA, p < 0.01). Taken together, these results imply that a saliva or a serum pellicle, and the carbon source in the environment, act a complex manner modulating Candida bioflim formation.

Acrylic Resins

A 4-year longitudinal study of the oral prevalence of enteric gram-negative rods and yeasts in Chinese children.

A 4-year longitudinal study of the oral prevalence of enteric gram-negative rods and yeasts in 116 Chinese primary school children in Hong Kong was conducted. The oral prevalence of enteric gram-negative rods for each consecutive year was 25.3%, 37.0%, 24.0% and 25.8% respectively, with a weighted mean of 27.9%. Enterobacteriaceae, which comprised 57% of all enteric gram-negative rods, were more common in children with no caries experience. The oral prevalence of yeasts for each consecutive year was 7.7%, 12.0%, 14.4% and 15.5% respectively, with a weighted mean of 12.5%. Candida albicans comprised 84% of all yeasts isolated. Oral yeast carriage was significantly associated with caries prevalence. While the oral prevalence of enteric gram-negative rods in primary school children in Hong Kong may be higher than in other parts of the world, repeated isolation of either enteric gram-negative rods or Candida spp. from individual children over the 4-year study period was rare, suggesting that carriage of these organisms is transient.

Candida

Association between carriage of oral yeasts, malnutrition and HIV-1 infection among Tanzanian children aged 18 months to 5 years.

The objective was to determine whether there is an association between carriage of oral yeasts, malnutrition and HIV-1 infection among Tanzanian children. A case-control study design within a cross-sectional study was used, and the outcome was carriage of oral yeasts. The exposure variables were malnutrition and HIV-1 antibody, and confounders to be adjusted for were age, sex, and breastfeeding. The study was carried out in Dar-es-Salaam, Tanzania, in two maternal and child health (MCH) clinics that offer routine medical checkups to all expectant mothers and children aged between 0 and 5 years in the catchment area. A total of 882 children aged between 18 months and 5 years participated. Smears from the tongue and buccal mucosa were examined for oral yeasts. Malnutrition was categorized according to standards on the MCH chart and World Health Organization/Centers for Disease Control (WHO/CDC) standards as weight-for-height (wasted), weight-for-age (underweight), and height-for-age (stunted). HIV-1 infection was determined by an enzyme-linked immunosorbent assay. Reactive sera were confirmed by Western Blot. About 27% of the children were slightly or severely malnourished according to standards on the MCH chart. According to WHO/CDC standards, 2.6% were wasted, 16.3% were underweight, and 29.6% were stunted. Fourteen (1.6%) were seropositive for HIV-1 antibody. Hyphal forms and blastospores were much more frequent among children infected with HIV-1 with odds ratios ranging from 3.8 (95% CI: 1.3;11.2) to 6.2 (95% CI: 2.1;18.4) depending on categorization of malnutrition. Malnutrition was a risk factor, too, albeit to a much lesser and insignificant degree. The study supports our previous findings that malnutrition may predispose to carriage of oral yeasts and subsequent infection. However, in this study population HIV infection was clearly the predominant risk factor.

AIDS-Related Opportunistic Infections