FUNGAL BIOFILMS AND DEVELOPMENT OF NEW ANTI-FUNGAL DRUGS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: TERMINATED
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 0232047
• Project No.: VT-HO1923
• Project Start Date: Oct 1, 2012
• Project End Date: Sep 30, 2015

Project Director
Johnson, D. I.

Recipient Organization
UNIVERSITY OF VERMONT
(N/A)
BURLINGTON,VT 05405

Performing Department
Molecular Genetics and Microbiology

Non Technical Summary
Biofilms caused by Candida albicans and other fungi lead to significant losses in the maple syrup industry by blocking/clogging maple sap plastic tubing. The studies described herein will allow us to examine the ability of biofilm inhibitors to block biofilms on this type of tubing and other abiotic surfaces. These studies will also give us insight into the mechanism by which one of the inhibitors, ETYA, is able to block these biofilms.

Animal Health Component

(N/A)

Research Effort Categories

Basic

60%

Applied

40%

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
501	4020	1102	30%
501	4020	1180	10%
712	4020	1030	30%
712	4020	1180	30%

Knowledge Area
501 - New and Improved Food Processing Technologies; 712 - Protect Food from Contamination by Pathogenic Microorganisms, Parasites, and Naturally Occurring Toxins;

Subject Of Investigation
4020 - Fungi;

Field Of Science
1180 - Pharmacology; 1030 - Cellular biology; 1102 - Mycology;

Keywords

biofilms

maple sap tubing

candida albicans

etya

Goals / Objectives
Our goal is to understand the mechanisms regulating biofilm formation by the model organism Candida albicans, and to develop new ways to inhibit biofilm formation on abiotic surfaces such as maple sap tubing. C. albicans is the model organism of choice when studying yeast biofilm formation, since the yeast Saccharomyces cerevisiae does not form significant biofilms. C. albicans biofilm formation depends on cells switching between budded and hyphal growth (budded-to-hyphal transition; BHT). Previously, we discovered 21 molecules that inhibited the BHT without killing cells. We identified the signaling pathways affected by some of these inhibitors, and importantly, we showed that three inhibitors, ETYA, buhytrinA, and CGP37157, blocked biofilm formation. The most potent inhibitor, ETYA, has several mechanisms of action in mammalian cells, but how it functions in C. albicans is unknown. Therefore, we will test two hypotheses: 1. ETYA blocks biofilm formation by inhibiting reactive oxygen species and/or oxygenase activity; and 2. ETYA, buhytrinA, and CGP37157 can be incorporated into maple sap plastic tubing and inhibit biofilm formation. These investigations may lead to the development of applied methods to block fungal biofilms on maple sap plastic tubing.

Project Methods
We will use established in vitro assays to examine the ability of the inhibitors ETYA, buhytrinA and CGP37157 to block biofilm formation on abiotic surfaces such as maple sap tubing. We will also use established biochemical and pharmacological techniques to examine the mechanism of action of ETYA, the most potent of the inhibitors. These studies will be performed by the PI, graduate student Todd Cramer, and several undergraduate researchers.

Progress 10/01/12 to 09/30/15

Outputs
Target Audience:Undergraduate students Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Four undergraduate researchers performed independent researchin the lab over the last year of funding. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Hypothesis 1. ETYA blocks biofilm formation by inhibiting reactive oxygen species and/or oxygenase activity. Using in vitro biofilm assays, we were able to show that the reactive oxygen species, hydrogen peroxide, had no effect on the biofilm-inhibiting activity of ETYA. We did show, however, that a calcium chelator, EDTA, did show synergistic effects with ETYA. Hypothesis 2. ETYA, buhytrinA, and CGP37157 can be incorporated into maple sap plastic tubing and inhibit biofilm formation. We showed that Candida albicansdoes form biofilms on maple sap tubing and thatETYA, but not buhytrinA orCGP37157,could inhibit the biofilm formation.

Publications

Progress 10/01/13 to 09/30/14

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? The goal of this project is to investigate the mechanisms by which the pathogenic yeast Candida albicans can form biofilms on abiotic surfaces, including map sap tubing. We have identified 21 small molecules that can block this biofilm formation on polystyrene microplates and are examining their role on tubing. Over the past year, we have successfully be able to induce biofilm formation on map sap tubing and contact lens and have shown that 3 small molecules can inhibit this process. We have also generated a quantitative assay to measure the amount of biofilms on these surfaces. We will be working on advancing the assay to different tubing and small molecules in the coming year, as well as examining the potential for incorporating the small molecules into the tubing using soaking methodology.

Publications

Progress 10/01/12 to 09/30/13

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? The goal of this project is to investigate the mechanisms by which the pathogenic yeast Candida albicans can form biofilms on abiotic surfaces, including map sap tubing. We have identified 21 small molecules that can block this biofilm formation on polystyrene microplates and are examining their role on tubing. Over the past year, we have successfully be able to induce biofilm formation on map sap tubing and contact lens and have shown that 3 small molecules can inhibit this process. We have also generated a quantitative assay to measure the amount of biofilms on these surfaces. We will be working on advancing the assay to different tubing and small molecules in the coming year, as well as examining the potential for incorporating the small molecules into the tubing using soaking methodology.

Publications