Jyy Yau

Biofilm-Forming Ability of Candida albicans Is Unlikely To Contribute to High Levels of Oral Yeast Carriage in Cases of Human Immunodeficiency Virus Infection

ABSTRACT An increased prevalence of candidal carriage and oral candidiasis is common in cases of human immunodeficiency virus (HIV) infection, and the reasons for this may include the enhanced ability of colonizing yeasts to produce biofilms on mucosal surfaces. The aim of the present study was therefore to examine the differences, if any, in the biofilm-forming abilities of 26 Candida albicans yeast isolates from HIV-infected individuals and 20 isolates from HIV-free individuals, as this attribute of yeast isolates from patients with HIV disease has not been examined before. Biofilm formation in microtiter plate wells was quantitatively determined by both the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT) reduction method and the crystal violet method. Although candidal biofilm formation could be quantitatively evaluated by either technique, the better reproducibility (P < 0.05) of the XTT reduction assay compared with that of the crystal violet method led us to conclude that the former is more reliable. There were no significant quantitative differences in biofilm formation between C. albicans isolates from HIV-infected patients and isolates from HIV-free individuals during in vitro incubation in a multiwell culture system over a period of 66 h. Three of eight host factors in the HIV-infected group were found to be associated with candidal biofilm formation. Thus, yeasts isolated from older individuals and those with higher CD4-cell counts exhibited decreased biofilm formation, while the findings for yeasts from individuals receiving zidovudine showed the reverse (P < 0.05 for all comparison). Our data indicate that attributes other than biofilm formation may contribute to the increased oral yeast carriage rates in cases of HIV infection.

In Vitro Method To Study Antifungal Perfusion in Candida Biofilms

ABSTRACT Antimycotic perfusion through Candida biofilms was demonstrated by a modification of a simple in vitro diffusion cell bioassay system. Using this model, the perfusion of three commonly used antifungal agents, amphotericin B, fluconazole, and flucytosine, was investigated in biofilms of three different Candida species (i.e., Candida albicans, Candida parapsilosis, and Candida krusei) that were developed on microporous filters. Scanning electron microscopy revealed that C. albicans formed a contiguous biofilm with tightly packed blastospores and occasional hyphae compared with C. parapsilosis and C. krusei, which developed confluent biofilms displaying structural heterogeneity and a lesser cell density, after 48 h of incubation on nutrient agar. Minor structural changes were also perceptible on the superficial layers of the biofilm after antifungal perfusion. The transport of antifungals to the distal biofilm-substratum interface was most impeded by C. albicans biofilms in comparison to C. parapsilosis and C. krusei. Fluconazole and flucytosine demonstrated similar levels of perfusion, while amphotericin B was the least penetrant through all three biofilms, although the latter appeared to cause the most structural damage to the superficial cells of the biofilm compared with the other antifungals. These results suggest that the antifungal perfusion through biofilm mode of growth in Candida is dependent both on the antimycotic and the Candida species in question, and in clinical terms, these phenomena could contribute to the failure of Candida biofilm-associated infections. Finally, the in vitro model we have described should serve as a useful system to investigate the complex interactions that appear to operate in vivo within the biofilm-antifungal interphase.

Phenotypic diversity of oral C. albicans isolated on single and sequential visits in an HIV-infected Chinese cohort.

HIV‐infected individuals maintain multiple oral C. albicans strains over time that are thought to undergo microevolution in terms of both phenotypic and genotypic features. To study this phenomenon, a 12‐month prospective study was conducted in a cohort of 16 HIV‐infected ethnic Chinese individuals with (A) and without (B) symptoms of oropharyngeal candidiasis to evaluate the phenotype distribution among oral C. albicans isolates during disease progression. Oral rinse samples were obtained and up to five C. albicans colony‐forming units were selected per each visit, during the one year period of multiple visits. The isolates were phenotyped using two commercially available biotyping kits, the API 20C system, API ZYM system, and a plate test for resistance to boric acid. A total of 261 C. albicans strains in group A were differentiated into 67 biotypes, while 42 biotypes were seen amongst the 182 isolates from group B. The major biotypes in the two groups were similar and were in decreasing order of prevalence J1R, J1S, J6S, J6R, J2S, K1S, J10R, K1R, and K6R; 48 different biotypes were seen in group A and 24 in group B, with some uniquely represented in each group, leading to a significant association between the prevalence of the biotypes J1S and J2S and symptomatic candidiasis (p<0.05). Taken together this study illustrates the wide phenotypic spectrum of oral C. albicans associated with HIV‐infection.