Pseudomonas aeruginosa Biofilm Formations & Bacteriophages Ps

Genevieve Johnson, Catherine Putonti

Loyola University Chicago

ABSTRACT: METHODS:
A biofilm is an aggregation of bacterial cells from the surrounding environment that are adhered to Strain Selection:
solid surfaces. The bacteriophage (virus that infects prokaryotic bacterial cells) is able to destroy and • Clinical isolates from urinary samples, including voided
prevent further growth of the bacterial biofilms. The goal of this project has been to grow and urine, catheterized urine, and kidney stones
characterize biofilms of clinical P. aeruginosa strains and test bacteriophage treatment.
• Strains of P. aeruginosa were chosen based on their ability
to form biofilms
• Specific strain was focused on: 8183, isolated from a
BIOFILMS: women with recurrent UTI
What Is A Biofilm?
• Aggregation of microbial cells enclosed
Growing the Biofilms:
• 37℃ in 20 mL tubes
in a polysaccharide matrix that is
• 35℃ & 0.25% Glucose
attached to a surface
• 6-Well Plates, 24-Well Plates, Glass Slides
• Thrive on moist or wet surfaces
Characterization:
Types of Biofilm Formations: • Crystal Violet Assays
• Surface attachment • Spectrophotometer- Biofilm Density
- More permanent through cell adhesion structures
Bacteriophage Treatment:
• Flocking
• SPAM & Spike!
- Cells anchor themselves to the matrix itself or to the
• Variable amounts of phage lysate
other nearby cells

Clinical Biofilms:
• Biofilms are associated with several symptoms/disease, RESULTS:
including urinary tract infections (UTIs)
• Antibiotics often cannot penetrate the polysaccharide Biofilms: Bacteriophage Treatment:
matrix • P. aeruginosa biofilms grew • Clear, initial lysis of biofilm by
sufficiently at 35℃ with added phage lysate
• Increasing resistance to antibiotics
added 0.25% glucose • Depletion of the biofilm
• Different biofilm structures are difficult to treat • Biofilms tended towards overall, but unable to measure
flocking rather than surface full lysis of the biofilm
attachment
Phage Treatment:
• Bacteriophage!
- Most abundant entity on planet, far surpassing the
population of bacteria CHALLENGES:
- Phage only attack and kill prokaryotic bacterial cells
• How do they work with biofilms? • Flocking made it a challenge to treat with antibiotics
• Growth conditions were stunted
- Phage can penetrate and kill cells within biofilms
- Plates dried out in aerated incubators
- Phage reproduce to attack the nearby bacterial cells - Biofilms did not grow well at 37℃

Acknowledgements: Thanks to Dr. Alan J Wolfe for providing the P. aeruginosa isolates and Bridget Brassil for her assistance. This work is supported by the Mulcahy Research Fellowship.