title: Alcohol dehydrogenase restricts the ability of the pathogen Candida albicans to form a biofilm on catheter surfaces through an ethanol-based mechanism.
abstract: Candida biofilms formed on indwelling medical devices are increasingly associated with severe infections. In this study, we used proteomics and Western and Northern blotting analyses to demonstrate that alcohol dehydrogenase (ADH) is downregulated in Candida biofilms. Disruption of ADH1 significantly (P = 0.0046) enhanced the ability of Candida albicans to form biofilm. Confocal scanning laser microscopy showed that the adh1 mutant formed thicker biofilm than the parent strain (210 microm and 140 microm, respectively). These observations were extended to an engineered human oral mucosa and an in vivo rat model of catheter-associated biofilm. Inhibition of Candida ADH enzyme using disulfiram and 4-methylpyrazole resulted in thicker biofilm (P < 0.05). Moreover, biofilms formed by the adh1 mutant strain produced significantly smaller amounts of ethanol, but larger amounts of acetaldehyde, than biofilms formed by the parent and revertant strains (P < 0.0001), demonstrating that the effect of Adh1p on biofilm formation is mediated by its enzymatic activity. Furthermore, we found that 10% ethanol significantly inhibited biofilm formation in vitro, with complete inhibition of biofilm formation at ethanol concentrations of >/=20%. Similarly, using a clinically relevant rabbit model of catheter-associated biofilm, we found that ethanol treatment inhibited biofilm formation by C. albicans in vivo (P < 0.05) but not by Staphylococcus spp. (P > 0.05), indicating that ethanol specifically inhibits Candida biofilm formation. Taken together, our studies revealed that Adh1p contributes to the ability of C. albicans to form biofilms in vitro and in vivo and that the protein restricts biofilm formation through an ethanol-dependent mechanism. These results are clinically relevant and may suggest novel antibiofilm treatment strategies.
authors.affiliation: Center for Medical Mycology, Department of Dermatology, Case Western Reserve University, 11100 Euclid Avenue, LKS-5028, Cleveland, OH 44106, USA.
authors.forename: Pranab K
authors.initials: PK
authors.lastname: Mukherjee
authors.forename: Sotohy
authors.initials: S
authors.lastname: Mohamed
authors.forename: Jyotsna
authors.initials: J
authors.lastname: Chandra
authors.forename: Duncan
authors.initials: D
authors.lastname: Kuhn
authors.forename: Shuqing
authors.initials: S
authors.lastname: Liu
authors.forename: Omar S
authors.initials: OS
authors.lastname: Antar
authors.forename: Ryan
authors.initials: R
authors.lastname: Munyon
authors.forename: Aaron P
authors.initials: AP
authors.lastname: Mitchell
authors.forename: David
authors.initials: D
authors.lastname: Andes
authors.forename: Mark R
authors.initials: MR
authors.lastname: Chance
authors.forename: Mahmoud
authors.initials: M
authors.lastname: Rouabhia
authors.forename: Mahmoud A
authors.initials: MA
authors.lastname: Ghannoum
mesh_terms: D000426:Alcohol Dehydrogenase / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D000818:Animals; D018441:Biofilms / Q000254:growth & development*; D002176:Candida albicans / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development* / Q000472:pathogenicity; D002404:Catheterization / Q000295:instrumentation; D004221:Disulfiram; D000431:Ethanol / Q000737:chemistry*; D017353:Gene Deletion; D040901:Proteomics; D011817:Rabbits; D051381:Rats; D013499:Surface Properties; D015854:Up-Regulation
chemical_list: D000431:Ethanol; D000426:Alcohol Dehydrogenase; D004221:Disulfiram