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Published on: January 16, 2019
Real-Time Visualization of Nutrient Media Impact on Pseudomonas aeruginosa Biofilm Development Using a Microfluidic
Niamh Corkery-Hayward1, Esther Karunakaran2, Melika Gul3
1School of Mechanical, Aerospace and Civil Engineering, University of Sheffield; ncorkery-hayward1@sheffield.ac.uk.
Abstract:
The pathogenic bacterium Pseudomonas aeruginosa is a major cause of numerous nosocomial infections, and its growing antimicrobial resistance has led to it posing a significant public health threat. This article presents a comprehensive protocol detailing the use of a microfluidic system for the real-time visualization and quantification of biofilm development in two key P. aeruginosa strains, PAO1 and PA14. The method employs optically transparent, multichannel microchannel plates to subject bacterial cultures to a continuous, steady flow of media, including tryptic soy broth (TSB) or modified minimal fastidious anaerobic broth (FAB) with varying carbon source concentrations, thereby mimicking conditions found in the clinical environments. Over a 24-h period, automated real-time imaging captures the growth and maturation of biofilms in the form of biofilm surface area coverage, thickness, and surface roughness in a highly reproducible manner. The experimental objective is to use the results to demonstrate that biofilm formation for both strains is significantly impacted by changes in nutrient media composition. The goal of this visualized protocol is to provide a method for researchers to study biofilm dynamics under steady laminar flow conditions, and the insights gained could be leveraged to develop alternative, non-antimicrobial strategies for eradicating early-stage P. aeruginosa biofilms in nosocomial settings.
Insights
This study visualizes Pseudomonas aeruginosa biofilm development using microfluidics. Nutrient media composition significantly impacts biofilm formation, offering insights for new eradication strategies against this resistant pathogen.
Area of Science:
- Microbiology
- Biotechnology
- Bioengineering
Background:
- Pseudomonas aeruginosa is a significant nosocomial pathogen with increasing antimicrobial resistance.
- Biofilm formation contributes to persistent infections and treatment challenges.
Purpose of the Study:
- To present a microfluidic protocol for real-time visualization and quantification of Pseudomonas aeruginosa biofilm development.
- To investigate the impact of nutrient media composition on biofilm formation in P. aeruginosa strains PAO1 and PA14.
Main Methods:
- Utilized optically transparent, multichannel microchannel plates for bacterial culture.
- Applied continuous, steady flow of tryptic soy broth (TSB) or modified minimal fastidious anaerobic broth (FAB) with varying carbon sources.
- Employed automated real-time imaging over 24 hours to capture biofilm dynamics.
Main Results:
- Successfully visualized and quantified biofilm surface area coverage, thickness, and roughness.
- Demonstrated that nutrient media composition significantly impacts biofilm formation in both PAO1 and PA14 strains.
- Achieved reproducible results under steady laminar flow conditions.
Conclusions:
- The developed microfluidic protocol enables detailed study of biofilm dynamics.
- Findings highlight the influence of nutritional environment on P. aeruginosa biofilm.
- Insights can inform the development of non-antimicrobial strategies for combating nosocomial infections.
