Impact of Brief Exposure to Lysozyme and Lactoferrin on Pathogenic Attributes of Oral Candida.

INTRODUCTION AND AIMS: Adhesion to buccal epithelial cells (BEC) and denture acrylic surfaces (DAS), germ tube (GT) formation, cell surface hydrophobicity (CSH), and haemolysin production are attributes associated with pathogenicity of Candida. Candida albicans and Candida dubliniensis are allied in causing oral candidosis. Lysozyme and lactoferrin exert antimicrobial activity on a range of oral microorganisms, including Candida. There is no information on the impact of brief exposure to lysozyme and lactoferrin on adhesion-related attributes and haemolysin production of aforementioned oral Candida isolates. Thus, we investigated the impact of lysozyme and lactoferrin on adhesion to BEC and DAS, GT formation, CSH, and haemolysin production of these isolates. METHODS: After exposure to lysozyme and lactoferrin for 1 hour, susceptibility to lysozyme and lactoferrin of 20 isolates each of C albicans and C dubliniensis isolates was determined following a 48-hour period of incubation. Candida cell suspensions, obtained from colony-forming units after this period, were assessed for adhesion to BEC and DAS, GT formation, CSH, and haemolysin production using in vitro assays. RESULTS: Exposure to lysozyme and lactoferrin significantly suppressed the ability of C albicans and C dubliniensis isolates to adhere to BEC and DAS, GT formation, CSH, and haemolysin production (P < 0.01 for all virulent attributes tested). CONCLUSIONS: These data provide a tantalising glimpse into the possibility that exposure to either lysozyme or lactoferrin, even for a brief period, would induce a sustainable antifungal effect by suppressing adhesion-related attributes and haemolysin production of these oral Candida species in vitro. Resistance to conventional antifungal agents has been reported in clinical isolates of Candida. The presence of such resistance indicates the need for possible alternative therapies to facilitate the management of oral candidosis. Further research on the pharmacodynamics of lysozyme and lactoferrin and their effects on candidal pathogenic attributes should be fostered, with the vision of developing novel topical antifungal drugs.

Impact of Cigarette Smoke Condensate on Adhesion-Related Traits and Hemolysin Production of Oral Candida dubliniensis Isolates.

BACKGROUND: Cigarette smoke is associated with higher oral Candida carriage and possible predisposition and increased susceptibility to oral candidal infection. Candida dubliniensis is associated with oral candidosis. Candidal adherence to buccal epithelial cells (BEC) and denture acrylic surfaces (DAS), germ tube (GT) formation, cell surface hydrophobicity (CSH) and hemolysin production are pathogenic traits of Candida. OBJECTIVES: The impact of exposure to cigarette smoke on the aforementioned pathogenic attributes of oral C. dubliniensis has not been studied. Hence, the impact of cigarette smoke condensate (CSC) on adhesion to BEC and DAS, GT formation, CSH and hemolysin production of 20 oral C. dubliniensis isolates after exposure to CSC for 24, 48 and 72 h was ascertained. METHODS: After preparation of the CSC, using an in-house smoking device, the Candida isolates were exposed to the CSC for 24, 48 and 72 h, by a previously described in vitro method. Thereafter, the adhesion to BEC and DAS, GT formation, CSH and hemolysin production of C. dubliniensis isolates was investigated by hitherto described in vitro assays. RESULTS: Exposure to CSC significantly increased the ability of C. dubliniensis oral isolates to adhere to BEC, DAS, GT formation, CSH and produce hemolysin following 24-h, 48-h and 72-h exposure periods to CSC (P < 0.001 for all attributes tested). CONCLUSIONS: Exposure of oral C. dubliniensis isolates to CSC may significantly promote in vitro adhesion traits and hemolysin production of these isolates, thereby augmenting its pathogenicity in vitro in the presence of cigarette smoke.

Comparison of the VITEK 2 antifungal susceptibility system with Etest using clinical isolates of Candida species / Comparación de los métodos de sensibilidad antifúngica Vitek 2 y Etest con cepas clínicas del género Candida

Background. Candida species are part of the normal human microbiota. However, in recent years, nosocomial bloodstream Candida infections have emerged as a significant problem ranking the fourth common cause of fungemia in intensive care units. Although microdilution methods are the ones recommended for susceptibility testing, they are difficult to undertake in the clinical practice. Thus, an automated commercially available test is ideal. Aims. To compare minimum inhibitory concentrations (MICs) obtained with the recently introduced Vitek 2 yeast susceptibility system card (AST-YS01) with Etest. Methods. 263 clinical Candida isolates representing six species were included in the study. Categorical agreements (CA) were assessed as described elsewhere. Results. Irrespective of the Candida species tested, the overall CA between Vitek 2 and Etest ranged between 66.7% and 100%. In general, Etest yielded lower MICs than Vitek 2. For Candida albicans, the CA between Vitek 2 and Etest was >95% for amphotericin B, voriconazole and flucytosine, but only 89% for fluconazole. With respect to Candida glabrata, the CA was between 97% and 100%. The major errors were with Candida krusei and flucytosine and Candida kefyr and amphotericin B. Candida tropicalis susceptibility for fluconazole by Vitek 2 reported more SDD and resistant strains than Etest. Candida parapsilosis showed 100% CA against all the four antifungals tested. No very major errors were detected between the two methods. Conclusions. Vitek 2 provided comparable results to Etest with quick turnaround for the testing of Candida species susceptibilities (AU)

Antecedentes. Candida forma parte de la microbiota habitual del ser humano. Sin embargo, en los últimos años, las candidemias hospitalarias se han convertido en un problema significativo en las unidades de cuidados intensivos al ocupar el cuarto lugar entre las fungemias. Puesto que los métodos de microdilución, recomendados para las pruebas de sensibilidad in vitro, son difíciles de realizar en la práctica clínica, las pruebas comerciales y automatizadas son las de uso ideal. Objetivos. Comparar las concentraciones mínimas inhibidoras (CMI) obtenidas por los métodos Vitek 2 (AST-YS01) y Etest. Métodos. Se utilizaron 263 cepas clínicas de Candida, pertenecientes a seis especies. Se evaluaron los acuerdos categóricos (AC) según lo ya descrito. Resultados. Con independencia de la especie de Candida, el AC general entre Vitek 2 y Etest osciló entre el 66,7 y el 100%. En general, Etest arrojó CMI más bajas que las de Vitek 2. Para Candida albicans el AC entre Vitek 2 y Etest fue > 95% con la anfotericina B, el voriconazol y la flucitosina, pero solo del 89% con el fluconazol. Con Candida glabrata el AC fue del 97-100%. Las mayores diferencias se registraron con Candida krusei y la flucitosina, y con Candida kefyr y la anfotericina B. Los valores de sensibilidad de Candida tropicalis con el fluconazol arrojaban más cepas SDD y resistentes con Vitek 2. El AC con Candida parapsilosis fue del 100% con todos los antifúngicos testados. No se observaron grandes diferencias entre los dos métodos. Conclusiones. Vitek 2 proporciona resultados comparables con los de Etest, con un tiempo rápido de respuesta respecto a las especies de Candida susceptibilidad (AU)

Comparison of the VITEK 2 antifungal susceptibility system with Etest using clinical isolates of Candida species.

BACKGROUND: Candida species are part of the normal human microbiota. However, in recent years, nosocomial bloodstream Candida infections have emerged as a significant problem ranking the fourth common cause of fungemia in intensive care units. Although microdilution methods are the ones recommended for susceptibility testing, they are difficult to undertake in the clinical practice. Thus, an automated commercially available test is ideal. AIMS: To compare minimum inhibitory concentrations (MICs) obtained with the recently introduced Vitek 2 yeast susceptibility system card (AST-YS01) with Etest. METHODS: 263 clinical Candida isolates representing six species were included in the study. Categorical agreements (CA) were assessed as described elsewhere. RESULTS: Irrespective of the Candida species tested, the overall CA between Vitek 2 and Etest ranged between 66.7% and 100%. In general, Etest yielded lower MICs than Vitek 2. For Candida albicans, the CA between Vitek 2 and Etest was >95% for amphotericin B, voriconazole and flucytosine, but only 89% for fluconazole. With respect to Candida glabrata, the CA was between 97% and 100%. The major errors were with Candida krusei and flucytosine and Candida kefyr and amphotericin B. Candida tropicalis susceptibility for fluconazole by Vitek 2 reported more SDD and resistant strains than Etest. Candida parapsilosis showed 100% CA against all the four antifungals tested. No very major errors were detected between the two methods. CONCLUSIONS: Vitek 2 provided comparable results to Etest with quick turnaround for the testing of Candida species susceptibilities.

Caspofungin-induced in-vitro post-antifungal effect and its impact on adhesion related traits of oral Candida dubliniensis and Candida albicans isolates.

Adhesion to buccal epithelial cells (BEC) and denture acrylic surfaces (DAS), germ tube (GT) formation and cell surface hydrophobicity (CSH) are all virulence traits involved in the pathogenicity of Candida. Post-antifungal effect (PAFE) also have a bearing on pathogenicity and virulence of Candida. Candida dubliniensis is associated with oral and systemic candidosis, which can be managed with caspofungin. There is no published information on caspofungin-induced PAFE and its impact on adhesion traits of C. dubliniensis isolates. Thus, the purpose of this investigation was to determine the in vitro duration of PAFE on 20 C. dubliniensis isolates following transient exposure to caspofungin. Furthermore the impacts of caspofungin-induced PAFE on adhesion to BEC and DAS, GT formation and CSH of these isolates were also determined. After establishing the minimum inhibitory concentration (MIC) of caspofungin, C. dubliniensis isolates were exposed to sub-lethal concentrations (×3 MIC) of caspofungin for 1 hr. Thereafter the duration of PAFE, adhesion to BEC and DAS, GT formation and CSH were determined by previously described in-vitro assays. MIC (µg/mL) of C. dubliniensis isolates to caspofungin ranged from 0.004 to 0.19. Caspofungin-induced mean PAFE on C. dubliniensis isolates was 2.17 hr. Exposure to caspofungin suppressed the ability of C. dubliniensis isolates to adhere to BEC and DAS, form GT and CSH by 69.97%, 71.95%, 90.06% and 32.29% (P < 0.001 for all), respectively. Thus, transient exposure of C. dubliniensis isolates to caspofungin produces an antifungal effect not only by suppressing its growth but also by altering its adhesion traits.