Inducing Dispersion of Filamentous Fungal Biofilms

Authors: Zara Lodi, Whitni Redman, Christopher Smyth, Jaclyn Printz

SUNY Campus: Binghamton University

Presentation Type: Oral

Location: UUW 324

Presentation #: 3

Timeslot: Session A 9:00-10:00 AM

Abstract: Filamentous fungi have become increasingly relevant in clinical environments, causing a rise in infections in immunocompromised patients and contact wearers due to their ability to form biofilm. One biofilm forming fungus, Fusarium, is responsible for causing mycotic keratitis, the fifth leading cause of blindness globally. Biofilms are group-mediated colonies of microbes surrounded by a protective layer called the extracellular polymeric substance (EPS). One proposed method to treat biofilm infections is by releasing cells from the biofilm by breaking down the EPS with the enzyme α-amylase, or inducing dispersion by decreasing c-di-GMP levels within the cells with sodium nitroprusside (SNP) to make the cells more vulnerable to antimicrobial agents. The aim of this study is to determine if Fusarium sp. biofilms can be dispersed. To determine when biofilm maturation occurred, a 96-well plate was inoculated with spore suspension and incubated at room temperature for 24, 48, and 72 hours. The biomass was stained with crystal violet, imaged, and eluted in ethanol and measured at an absorbance of 570 nm. Biofilm maturation was determined to occur at 48 hours of incubation. Next, a dispersion assay was performed by growing biofilms for 48 hours in a 24-well plate at room temperature. The biofilms were then treated with α-amylase, SNP, or media control for two hours. The supernatant and biofilm were collected to determine percent dispersed. Our preliminary findings suggest that α-amylase and SNP can release cells from filamentous fungal biofilms, suggesting a potential clinical application.