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Technologies for Emerging Infectious Diseases

Technologies for Emerging Infectious Diseases - Poster Session A

Poster K12 - Application of Fungal Extracts to Inhibit Biofilm Formation on Medically Implantable Devices- Team Fungi, Anju Meda (Presentor)

Thursday, October 24, 2024

10:00 AM - 11:00 AM EST

Location: Exhibit Hall E, F & G

Presenting Author(s)

Anju Meda, Banneker Key Scholar ;Gemstone Honors Program

Undergraduate Student Researcher
University of Maryland College Park
Germantown, Maryland, United States

Co-Author(s)

CG

Catalina Gibney

Undergraduate Student Researcher
Gemstone Honors College- University of Maryland College Park, United States
LC

Leah Crowley

undergraduate student researcher
Gemstone Honors College- University of Maryland College Park, United States
MK

Matthew Kong (he/him/his)

Undergraduate Student Researcher
Gemstone Honors College- University of Maryland College Park, United States
RC

Ranita Chowdhury

Undergraduate Student Researcher
Gemstone Honors College- University of Maryland, United States

Presenting Author(s)

AC

Andrew Cardillo

Undergraduate Student Researcher
Gemstone Honors College- University of Maryland, United States

Co-Author(s)

BG

Beyza Gul

Undergraduate Student Researcher
Gemstone Honors College- University of Maryland, United States

Presenting Author(s)

Annette Eldo

Undergraduate Student Researcher
Gemstone Honors College- University of Maryland, United States

Co-Author(s)

LB

Lili Bao

Undergraduate Student Researcher
Gemstone Honors College- University of Maryland, United States
SM

Sumangal Myers

Undergraduate Student Researcher
Gemstone Honors College- University of Maryland, United States

Primary Investigator(s)

MP

Myles Poulin

Faculty Mentor
Gemstone Honors College- University of Maryland, United States

Introduction: Biofilms are defined as complex colonies of bacteria that can grow on medically implantable device surfaces and typically have antibiotic-resistant properties. 50~70% of healthcare-associated infections are derived from microbial growth on implantable medical devices (VanEpps & Younger, 2016). Currently, there are no commercially available FDA-approved or EPA-certified biofilm sterilants that are biocompatible for medically implantable devices. Team Fungi aims to study potential applications of fungal extracts as a sterilant as previous literature indicates fungal extracts to displace biofilm function and effectively inhibit biofilm growth. In this regard, Team Fungi aims to prevent medically- and industrially-relevant biofilm formation using fungal extracts to address the uptick in antibiotic resistance and create a more equitable and low-cost treatment. Bioactive metabolites isolated from mushrooms are rising in popularity in medical research due to their antimicrobial, antiviral, and immunomodulatory properties. Through the isolation of extracts derived from Turkey Tail and Shiitake mushrooms, this research seeks to find a sustainable, safe, and accessible method to treat biofilm infections originating from medical settings and medically implanted devices to address the rise in antibiotic resistance. Current results have shown that the mean absorbance decreased as dilution of the treatment solution increased, indicating biofilm formation was being inhibited by the addition of fungal extracts. We hope to apply our research to treat biofilm extractions that form on medical devices such as catheters, pacemakers, and prosthetics.

Materials and

Methods: --MATERIALS--
- Dried Turkey Tail and Shiitake Mushroom
- Soxhlet Extraction Apparatus
- Coffee Grinder
- Ultrasonic Extraction Apparatus
- 96 Well Plate
- PBS
- S. Epidermidis Glycerol Stock + Loop
- Crystal Violet Stain
- Ethanol
- Methanol
- Round Bottom Flask
- Baffled and Conical Flask
- Tryptic Soy Broth
- Plate Reader
- Shaker
- Rotovap
- Centrifuge
- Vacuum Filter Apparatus
- P-1000 Pipette

--METHODOLOGY--
First, the extraction of bioactive compounds in our mushroom extracts (that we plan to later identify with characterization assays like Mass-spectrometry and IR) are isolated with either Soxhlet or Ultrasonic extractions to isolate fungal metabolites from Turkey Tail and Shiitake mushrooms. 0.5g of ground mushroom is dissolved in 42 mL of organic solvent and extracted for 18-24 hrs to obtain an extract. The extract is then vacuum filtered and the organic solvent is dried via Rotovap. Then TSB cultures with glycerol stock are prepared, autoclaved, and inoculated with Staph Epidermidis to form biofilm cell cultures. The extract is resuspended with 3.5 mL of PBS Buffer and serially diluted with PBS buffer into overnight culture following dilution scheme and plated on a 96-well plate. Then media is aspirated out and cells are fixed with methanol and stained with crystal violet to quantify and stain the biomass. The plate is read on a plate reader and absorbance vs. dilution factors are plotted with error bars for the control and experimental wells to effectively quantify the inhibition of biofilm mass formation.

Results, Conclusions, and Discussions: To further investigate the effectiveness of our isolated mushroom extracts, the bioactive metabolites from Turkey Tail and Shiitake mushrooms were extracted using ultrasonic-assisted methods and Soxhlet extraction techniques. Then, the mushroom extracts were then analyzed by completing a microtiter plate assay and staining the cells to read the absorbance and determine if inhibition was achieved. Current results have shown that the mean absorbance decreased as dilution of the treatment solution increased, indicating biofilm formation was being inhibited by the addition of fungal extracts. Soxhlet crude extract exhibited 72% inhibition in biomass growth in a dose-dependent manner while the ultrasonic extract had no defining trend of inhibition. We hope to apply our research to treat biofilm extractions that form on medical devices such as catheters, pacemakers, and prosthetics.

Thus far in our experimentation, we have utilized water-based extracts in Soxhlet and ultrasonic methods to obtain the fungal extracts. We hope to utilize organic-based extracts in the future to maximize our yield and acquire pure samples. Additionally, we hope to fractionate fungal extracts to characterize the bioactive compounds within our extracts that exhibit positive signs of biofilm inhibition. Lastly, we hope to conduct cytotoxicity assays to determine the sterilant success of our isolated extracts on bacterial strains prevalent in hospital infections.

Acknowledgements (Optional): We would like to thank our mentor Dr. Myles Poulin, librarian Isabella Baxter, and Gemstone Staff: Dr. Allison Lansverk, Leslie Lizama, Dr. David Lovell, and Brianna Lucas.
In addition, we thank LaunchUMD, the Do Good Institute and University Libraries Award for Outstanding Gemstone Team for supporting our research.