Biomedical subjects

L P Samaranayake

Publications and source records attributed to L P Samaranayake.

At least 55 records · Page 3Linked to original sources

PubMed2002

Candida glabrata, an emerging fungal pathogen, exhibits superior relative cell surface hydrophobicity and adhesion to denture acrylic surfaces compared with Candida albicans.

Oral candidosis is a common opportunistic infection in debilitated individuals and Candida glabrata is the second most frequently isolated species from this condition, after Candida albicans. Candidal adherence to various biological or non-biological surfaces is considered a prerequisite for colonization, and pathogenesis of candidal infections, and their relative cell surface hydrophobicity (CSH) is likely to be a possible contributory force involved in this process. Whereas a large body of data on the latter features of C. albicans is available, there is surprisingly little information on C. glabrata. As a comprehensive database on the relative adhesion and CSH of Candida spp. is instructive and useful, we investigated in vitro the latter attributes of 34 oral isolates of C. glabrata and 15 isolates of C albicans. There were remarkable intraspecies differences in both the CSH and the adhesive ability of C. glabrata strains (p < 0.001). Compared with C. albicans, C glabrata demonstrated a four-fold greater CSH value (30.63 +/- 11.20% vs 7.23+/-3.56%, p < 0.0001) and a two-fold greater tendency to adhere to denture acrylic surfaces (75.18 +/- 39.96 vs 30.36+/-9.21, p < 0.0001). A significant positive correlation between CSH and adhesion was also noted for both C. glabrata (r=0.674, p < 0.0001) and C. albicans ( r = 0.636, p < 0.05). When the effect of different incubation conditions on the relative CSH and adherence of C. glabrata was examined, CSH and the adherence to acrylic surfaces of four of six C. glabrata isolates were significantly affected by a reduction of the culture temperature (from 37 degrees C to 25 degrees C). A positive relationship also emerged when the temperature-induced variations in the adherence values were correlated with their relative CSH. These data provide hitherto unavailable archival information on important pathogenic attributes of the two most common oral Candida species that may help explain their predominance in this milieu.

Acrylic Resins↗

PubMed2002

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.

Adult↗

PubMed2002

Bacterial and yeast flora of root surface caries in elderly, ethnic Chinese.

OBJECTIVES: Root caries is emerging as a significant problem in the middle aged and elderly. As little data is available on the microbiology of root caries in Chinese cohorts, we evaluated 30 such lesions in elderly, institutionalized, ethnic Chinese. METHODS: Samples of carious dentine were aseptically taken from root caries lesions of 18 subjects (five males and 13 females, mean age 79.67 +/- 8.57). The cultivable bacteria, both aerobic and anaerobic, were isolated and identified using standard methods and commercial identification kits. The yeasts were isolated on Sabouraud's agar and identified using the API system. RESULTS: The main findings were: (1) of the total isolates, 91.09% were Gram-positive and 8.91% were Gram-negative microorganisms; (2) the proportions of cocci and rods were 36.68 and 63.31%, respectively; (3) the predominant groups of organisms isolated were Streptococcus spp., Lactobacillus spp., Staphylococcus spp. and Actinomyces spp. with isolation frequencies of 100, 90.00, 73.33 and 63.33%, respectively; (4) the isolation frequency of yeasts belonging to Candida spp. (63.33%) was notably high although the proportion of yeasts within each sample was low (0.01%). Candida dubliniensis, a newly identified yeast species particularly prevalent in HIV infection, comprised 14.29% of yeasts. CONCLUSIONS: This study provides baseline information on the microbiologic features of root caries in the elderly, ethnic Chinese. Whilst our data on the most predominant bacteria isolated in root caries concur with those from other regions of the world the isolation of C. dubliniensis from these lesions has not been documented before.

Actinomyces↗

PubMed2002

Impact of lysozyme and lactoferrin on oral Candida isolates exposed to polyene antimycotics and fluconazole.

OBJECTIVE: To assess the antifungal activity of lysozyme and lactoferrin on 10 oral isolates each of Candida albicans and Candida tropicalis following their brief exposure (I h) to subtherapeutic concentrations of two polyene drugs - nystatin, amphotericin B, and an azole -fluconazole. METHODS: Yeasts were sequentially exposed to subtherapeutic concentrations of antifungals and then to either lysozyme or lactoferrin and the viability evaluated by quantifying colony-forming units. RESULTS: The exposure of both C. albicans and C. tropicalis isolates to all three antifungal agents significantly increased their susceptibility to lysozyme (P < 0.05) but not to lactoferrin. Exposure to the two polyene drugs had a lesser impact on the lysozyme susceptibility of both Candida species compared with the azole drug. Both interspecies and intraspecies sensitivity to lysozyme was noted and C. albicans was less susceptible than C. tropicalis. CONCLUSIONS: Lysozyme, in addition to being a potent natural antifungal agent, may act synergistically with the studied antimycotics.

Amphotericin B↗

PubMed2002

Fungal infections associated with HIV infection.

Oral candidiasis is perhaps the commonest infection seen in HIV disease. The aim of this workshop was to provide a sketch of the multifarious aspects of the disease from a global perspective. To this end the panellists addressed issues such as the virulence of Candida, emergence of antifungal resistance, management of candidiasis and other exotic, oral mycotic diseases. An all-pervasive theme was the dramatic differences in the management of fungal infections consequential to the availability (or the lack) of anti-HIV drugs in the developed and the developing world. Further, the social stigmata associated with the HIV disease in many developing regions in Africa and Asia appears to modify the therapeutic strategies. Additionally, the lesser-known regional variations in the disease manifestations and therapeutic approaches were stark. Further work is direly needed to address these issues.

AIDS-Related Opportunistic Infections↗

PubMed2002

Differences in Candida albicans adhesion to intact and denatured type I collagen in vitro.

An inhibition assay of Candida albicans adhesion to gelatin-immobilized membranes was compared with that to intact type I collagen-immobilized membranes using an arginine-glycine-aspartic acid (RGD) containing peptide. As compared with a protein-free membrane, gelatin and collagen significantly enhanced the adherence of C. albicans. The adhesion of the yeast to gelatin was significantly inhibited by the RGD peptides, but not by arginine-glycine-glutamic acid (RGE) peptides. In contrast, attachment to collagen was not inhibited by RGD peptides. These results suggest that the RGD sequence of gelatin and the integrin-like proteins of yeasts may be involved in adherence.

Adhesiveness↗

PubMed2002

Case Report. Oro-facial manifestations of Penicillium marneffei infection in a Thai patient with AIDS.

Penicillium marneffei infection is prevalent in South-East Asia and Southern China and has been considered an acquired immunodeficiency syndrome (AIDS)-defining disease. This report focuses on the oral and facial manifestations of P. marneffei infection in a 28-year-old Thai male patient with AIDS. The clinical, mycological and ultrastructural features are described.

AIDS-Related Opportunistic Infections↗

PubMed2002

High oral prevalence of Candida krusei in leprosy patients in northern Thailand.

Although Candida albicans is the most common human yeast pathogen, other Candida species such as C. krusei are now recognized as emerging agents, especially in patients with human immunodeficiency virus (HIV) disease. C. krusei is inherently resistant to the widely used triazole antifungal fluconazole and poses therapeutic problems, especially in systemic candidiasis. In a surveillance study of leprosy patients (with arrested or burnt-out disease) in a leprosarium in northern Thailand, we found a rate of oral carriage of C. krusei (36%) significantly (P < 0.05) higher than that for a healthy control group (10%). Among the Candida-positive patients, 16 of 35 (46%) carried C. krusei, while C. albicans was the second most common isolate (12 of 35 patients; 34%). The corresponding figures for the control group were 2 of 13 (15%) and 6 of 13 (46%), respectively. Studies of the antifungal resistance of the C. krusei isolates from patients indicated that all except one of the isolates were resistant to fluconazole, two isolates were resistant to ketoconazole, and all isolates were sensitive to amphotericin B. Evaluation of their genetic profiles by randomly amplified polymorphic DNA analysis with three different primers and subsequent analysis of the gel profiles by computerized cluster-derived dendrograms revealed that the C. krusei isolates from patients belonged to 10 disparate clusters, despite the origin from a single locale. These nascent findings indicate an alarmingly high prevalence of a Candida species resistant to a widely used antifungal in a part of the world where HIV disease is endemic.

Aged↗

PubMed2001

Phenotypes and randomly amplified polymorphic DNA profiles of Candida albicans isolates from root canal infections in a Finnish population.

A total of thirty-seven Candida albicans isolates from root canal infections in a Finnish population were subtyped using phenotypic and genotypic methods. A previously described biotyping method based on the presence of five different enzymes, assimilation of eleven different carbohydrates and boric acid sensitivity of the yeasts was used to determine the phenotype. Commercially available API ZYM and API 20 C test kits were used to determine the presence of enzymes and assimilation of carbohydrates. The sensitivity of the isolates to boric acid was tested by their ability to grow on yeast-nitrogen-agar with incorporated boric acid (1.8 mg. ml(-1)). Combination of the tests revealed a total of 14 different phenotypes. The majority of the isolates, 26 strains, were classifiable into three major phenotypes: 16 isolates (43.2%) belonged to phenotype A1R, six (16.2%) to A1S and four (10.8%) to B1S. The remaining 11 phenotypes represented only a single isolate each. The randomly amplified polymorphic DNA profiles were used to determine the genotypes. For this purpose two different primers, RSD6 and RSD12 were used to develop a combination randomly amplified polymorphic DNA profile for each isolate. Altogether 31 genotypes were noted among the 37 isolates, of which only three pairs of isolates presented with congruent phenotypic and genotypic profiles. The heterogeneity of both the phenotypic and randomly amplified polymorphic DNA profiles of C. albicans isolates from root canal infections is akin to previous reports from other oral and non-oral sources in different geographic locales.

Bacterial Typing Techniques↗

PubMed2001

Oral colonization of aerobic and facultatively anaerobic gram-negative rods and cocci in irradiated, dentate, xerostomic individuals.

This study aimed at investigating the oral colonization of aerobic and facultatively anaerobic gram-negative rods and cocci in head- and neck-irradiated, dentate, xerostomic individuals. Subjects were recruited from a nasopharyngeal carcinoma clinic and were segregated into group A: <60 years (n=25, 48+/-6 years, 5+/-5 years post-irradiation) and group B: >or=60 years (n=8, 67+/-4 years, 2+/-2 years post-irradiation) and were compared with age- and sex-matched normal individuals, group C: <60 years (n=20, 44+/-12 years) and group D: >or=60 years (n=10, 70+/-3 years). Selective culture of the oral rinse samples was carried out to isolate, quantify and speciate (using API 20E kit) aerobic and facultatively anaerobic gram-negative rods and cocci recovery. All test subjects were put under comprehensive oral and preventive care for 3 months, and 12 group A and 5 group B subjects were recalled for reassessment of aerobic and facultatively anaerobic gram-negative rods and cocci colonization. All identical isolates, pre- and post-hygienic care, were phenotypically (Vitek, Hazelwood, MA and antibiogram profile) and genotypically (pulsed-field gel electrophoresis) evaluated. The aerobic and facultatively anaerobic gram-negative rods and cocci isolated from the first round oral rinse samples included: Acinetobacter, Neisseria, Chryseomonas, Flavimonas, Pseudomonas, Citrobacter, Enterobacter, Escherichia, Klebsiella, Flavobacterium and Weeksella species. The aerobic and facultatively anaerobic gram-negative rods and cocci isolation rate was high for irradiated individuals, and they were 64/25% and 100/80% for groups A/C and B/D, respectively. Recovery of aerobic and facultatively anaerobic gram-negative rods and cocci and Klebsiella pneumoniae subsp. pneumoniae in oral rinse samples were found to be significantly more prevalent in the irradiated subjects (groups A and B). Enterobacteriaceae were more frequently isolated from oral rinse samples of aged irradiated subjects (group B vs D, P<0.05), where the quantity of Citrobacter freundii (colony-forming units/ml oral rinse) was also significantly elevated. The isolation rate of aerobic and facultatively anaerobic gram-negative rods and cocci after hygienic care remained unchanged; 3 of 12 and 3 of 5 of the recalled subjects from groups A and B, respectively, harbored same aerobic and facultatively anaerobic gram-negative rods and cocci species. However, only two pairs of K. pneumoniae subsp. pneumoniae, sequentially isolated from same patients in group B, were found to be identical by pulsed-field gel electrophoresis. This may be due to reinfection of the microbes from the same source or permanent colonization. In conclusion, irradiation-induced xerostomia seems to favor frequent, repeated, transient intraoral colonization of aerobic and facultatively anaerobic gram-negative rods and cocci.

Acinetobacter↗

PubMed2001

Predominant cultivable subgingival microbiota of healthy and HIV-infected ethnic Chinese.

Although clinically distinctive types of periodontal disease are known to be associated with HIV infection, the pathogenesis remains unclear. In the present study, the subgingival microflora of 21 HIV-infected and 11 HIV-free ethnic Chinese were studied using the direct microscopic and anaerobic culture methods. Motile curved rods were found to be three times higher in the HIV-infected group under direct microscopy. Otherwise, there were no significant differences between the diseased and healthy groups when analyzed either in relation to the morphotype distribution or Gram stain morphology or oxygen tolerance. The most common bacteria isolated were Capnocytophaga species followed by Prevotella loescheii, Streptococcus sanguinis, Lactobacillus spp. and Fusobacterium spp. Although there were 20 bacterial species that were strictly limited to the HIV-infected group, and 5 limited to the healthy group, none of the species was a predominant isolate in either group (p>0.05). These findings agree with previous studies which report that subgingival bacteria of HIV-infected individuals (with or without periodontal disease) are similar to those found in healthy HIV-free individuals (with or without periodontal disease). Further work examining the subgingival microflora from patients with specific and/or severe forms of HIV-associated periodontal disease is required to shed light on the pathogenesis of this complex clinical entity.

Adult↗

PubMed2001

The effect of oral bacteria on Candida albicans germ-tube formation.

A total of eight bacterial isolates belonging to six species, and a select group of 12 oral Candida albicans isolates, were used to study the effect of bacteria on germ-tube formation. Briefly, each bacterial suspension (10(5-6) cells/ml) was mixed with a C. albicans suspension (10(7) cells/ml) and incubated at 37 degrees C for 90 min with bovine serum, and the percentage germ-tube-positive Candida cells was quantified using a haemocytometer, under light microscopy. In general, out of eight bacteria, Streptococcus sanguis SK21A, Streptococcus salivarius SK56, Escherichia coli ATCC 25922, and S. salivarius OBU3 suppressed germ-tube formation to varying degrees, with different C albicans isolates. Porphyromonas gingivalis Pg 50, Lactobacillus casei ATCC 7469 and Prevotella intermedia OBU4 elicited significant enhancement of germ-tube formation, whereas S. sanguis OBU 2 had no effect. E. coli ATCC 25922 was the only organism to show statistically significant suppression of germ-tube formation (p=0.0312). A significant increase in the germ tube production of C. albicans isolated from HIV-infected compared with HIV-free individuals was also noted. The current results tend to suggest that commensal and transient oral bacterial populations may selectively influence the differential expression of germ-tube-forming ability of C. albicans isolates.

Candida albicans↗

PubMed2001

Post-antifungal effect of polyene, azole and DNA-analogue agents against oral Candida albicans and Candida tropicalis isolates in HIV disease.

Oropharyngeal candidiasis (OPC) is the most frequent AIDS-associated opportunistic infection, as up to 90% of HIV-infected individuals suffer at least one episode during the course of their disease. Various in vivo and in vitro procedures have been used to assess the effectiveness of antifungal agents used in HIV infection. In the present study, we evaluated in vitro the minimum inhibitory concentration (MIC) and the post-antifungal effect (PAFE) of two polyenes, two azoles and one DNA-analogue against 10 oral isolates of Candida albicans and 10 of Candida tropicalis, all from HIV-infected individuals, in order to obtain basic data on the pharmacodynamics of these drugs. One-hour exposure to twice the MIC of all the drugs, except fluconazole, elicited a consistently high PAFE in both Candida species. Furthermore, the PAFE elicited by the antifungals (except fluconazole) was significantly prolonged for C. tropicalis compared with C. albicans. This speedy recovery of C. albicans isolates exposed to transient low concentrations of antifungals appeared to reflect its virulence compared with lesser potent species, such as C. tropicalis. Taken together, the current data, while confirming the existence of PAFE in a non-albicans species of Candida, also provide further clues for the recalcitrance of C. albicans species in the face of antifungal therapy for oropharyngeal candidiasis.

AIDS-Related Opportunistic Infections↗

PubMed2001

Heterogeneity in antifungal susceptibility of clones of Candida albicans isolated on single and sequential visits from a HIV-infected southern Chinese cohort.

The increased frequency and severity of candidal infections in human immunodeficiency virus (HIV)-infected individuals has prompted the wide use of antifungals, such as amphotericin B, ketoconazole, and fluconazole, resulting in the emergence of drug-resistant strains of Candida albicans. To study this phenomenon in an ethnic Chinese cohort, we isolated multiple colonies of Candida from the oral cavities of 16 HIV-infected patients on single and subsequent sequential visits over a period of 12 months. Ten of the 16 patients had sporadic episodes of oropharyngeal candidiasis (Group A), while the remainder were asymptomatic with respect to this condition (Group B). Oral rinses were collected and immediately processed in the laboratory for the isolation of C. albicans in a standard manner. A total of 433 C. albicans isolates were tested for their susceptibility to amphotericin B, ketoconazole and fluconazole by an agar diffusion method using the commercially available E-test. All tested isolates demonstrated variable susceptibility to amphotericin B, ketoconazole and fluconazole. The minimum inhibitory concentration (MIC) of the isolates for amphotericin B, ketoconazole and fluconazole ranged from <0.002-1.5 microg/ml, <0.002-4.0 microg/ml and <0.016-32 microg/ml, respectively. Sequential isolates of a few patients demonstrated variable susceptibility to all the antifungals, and no discernible MIC pattern emerged either in group A or B over time. Interestingly, significant variation in antifungal susceptibility was also noted in isolates obtained from the same patient on a single visit. Sequential yeast isolates in 9 of 16 patients (56%) demonstrated significant differences in MIC within and between visits for both amphotericin B and ketoconazole, while a lower percentage--44%(7/16)--exhibited this trait for fluconazole. Our study demonstrates the diversity in antifungal susceptibility in either commensal or "infective" oral strains of C. albicans in HIV disease, and shows the need for vigilance for the emergence of resistant strains, and for frequent antifungal susceptibility studies.

AIDS-Related Opportunistic Infections↗

PubMed2001

Adhesion of Candida parapsilosis to epithelial and acrylic surfaces correlates with cell surface hydrophobicity.

We investigated in vitro adherence of 24 isolates of Candida parapsilosis and 12 isolates of Candida albicans with regard to their relative cell-surface hydrophobicity (CSH), adherence to human buccal epithelial cells (BEC) and acrylic surfaces. There was no significant interspecies difference in the relative adherence of C. parapsilosis and C. albicans isolates to BEC, although the former demonstrated a tendency for increased adherence. However, a significant intra-species variation in adherence among isolates of C. parapsilosis (P < 0.0001) to BEC, but not of C. albicans was noted. The superficial isolates of C. parapsilosis demonstrated a higher avidity (33%) to BEC than the systemic isolates. On regression analysis a significant positive correlation between C. parapsilosis adherence to BEC and denture acrylic surfaces was noted (r = 0.45, P = 0.02). Similarly, buccal cell adherence correlated strongly with CSH of C. parapsilosis (r = 0.63, P = 0.0008). These results shed further light on the intimate relationship between adherence and CSH in candidal colonization and imply that both C. parapsilosis and C. albicans are equally potent in colonizing mucosal surfaces with respect to the attributes investigated.

Acrylic Resins↗

PubMed2001

Experimental oral candidiasis in animal models.

Oral candidiasis is as much the final outcome of the vulnerability of the host as of the virulence of the invading organism. We review here the extensive literature on animal experiments mainly appertaining to the host predisposing factors that initiate and perpetuate these infections. The monkey, rat, and mouse are the choice models for investigating oral candidiasis, but comparisons between the same or different models appear difficult, because of variables such as the study design, the number of animals used, their diet, the differences in Candida strains, and the duration of the studies. These variables notwithstanding, the following could be concluded. (i) The primate model is ideal for investigating Candida-associated denture stomatitis since both erythematous and pseudomembranous lesions have been produced in monkeys with prosthetic plates; they are, however, expensive and difficult to obtain and maintain. (ii) The rat model (both Sprague-Dawley and Wistar) is well proven for observing chronic oral candidal colonization and infection, due to the ease of breeding and handling and their ready availability. (iii) Mice are similar, but in addition there are well characterized variants simulating immunologic and genetic abnormalities (e.g., athymic, euthymic, murine-acquired immune deficiency syndrome, and severe combined immunodeficient models) and hence are used for short-term studies relating the host immune response and oral candidiasis. Nonetheless, an ideal, relatively inexpensive model representative of the human oral environment in ecological and microbiological terms is yet to be described. Until such a model is developed, researchers should pay attention to standardization of the experimental protocols described here to obtain broadly comparable and meaningful data.

Animals↗

PubMed2001

Effects of two different growth media on the postantifungal effect induced by polyenes on Candida species.

There are no data on the effects of different growth media on polyene-induced postantifungal effect (PAFE) in Candida species. Hence, the nystatin- and amphotericin B-induced PAFEs in six Candida species (26 isolates) grown in Sabouraud's dextrose broth (SAB) and RPMI broth were evaluated, following limited exposure to the MICs of the two polyenes, using an automated turbidometric method. For nystatin, PAFE varied between 1.88 and 4.87 h in SAB and 0.66 and 6.89 h in RPMI, and for amphotericin B, the equivalent values were 3.13 to 10.98 h in SAB and 0.97 to 7.01 h in RPMI. These highly significant (P < 0.001) variations in the PAFE with both drugs, noted with most Candida strains grown in different media, call for standardization of intralaboratory methodology in measuring this parameter in order to obtain universally comparable data.

Amphotericin B↗

PubMed2001

Candida species exhibit differential in vitro hemolytic activities.

A total of 80 Candida isolates representing 14 species were examined for their respective responses to an in vitro hemolytic test. A modification of a previously described plate assay system where the yeasts are incubated on glucose (3%)-enriched sheep blood agar in a carbon dioxide (5%)-rich environment for 48 h was used to evaluate the hemolytic activity. A group of eight Candida species which included Candida albicans (15 isolates), C. dubliniensis (2), C. kefyr (2), C. krusei (4), C. zeylanoides (1), C. glabrata (34), C. tropicalis (5), and C. lusitaniae (2) demonstrated both alpha and beta hemolysis at 48 h postinoculation. Only alpha hemolysis was detectable in four Candida species, viz., C. famata (3), C. guilliermondii (4), C. rugosa (1), and C. utilis (1), while C. parapsilosis (5) and C. pelliculosa (1) failed to demonstrate any hemolytic activity after incubation for 48 h or longer. This is the first study to demonstrate the variable expression profiles of hemolysins by different Candida species.

Animals↗