Evaluating the Effectiveness of Chemical Agents on Biofilm Formation in Escherichia coli Utilizing Quantitative and Qualitative Techniques
Faculty Mentor
Vinaya Sampath
Area of Research
Microbiology
Major
Forensic Science and Biology (with a Pre-Medical Sciences Concentration)
Description
INTRODUCTION: Gram-negative bacteria produce biofilms that contribute to their antibiotic resistance, which makes infections harder to treat.
METHOD: DH5-alpha strains of Escherichia coli were grown in Luria-Bertani broth, inoculated in 24-well polystyrene plates containing coverslips, and incubated at 37C for one week. Biofilm disruption techniques were followed utilizing 0.05% vs 0.1% Tween-20, Triton X-100, and Sodium Dodecyl Sulfate (SDS). Crystal violet was used to stain the bacteria in the biofilms, and qualitative observations of the detergent’s impact were made under the microscope.
RESULTS: 0.1% concentrations of Tween-20 and SDS worked effectively at disrupting the biofilm than 0.05% concentrations. However, we observed that 0.05% concentrations of Triton X-100 worked better than 0.1% concentrations.
DISCUSSION/CONCLUSION: Future work will use fluorescent dyes, such as CFSE and Propidium iodide, to quantify the proportion of live vs dead bacteria to get more insight into the composition of bacteria in the biofilms. Analysis will also be done to evaluate the effectiveness of alternative anti-biofilm strategies, such as the use of extracellular enzymes and quorum quenching, and how to use them in tandem with antibiotic delivery agents to attack gram-negative bacteria.
Evaluating the Effectiveness of Chemical Agents on Biofilm Formation in Escherichia coli Utilizing Quantitative and Qualitative Techniques
INTRODUCTION: Gram-negative bacteria produce biofilms that contribute to their antibiotic resistance, which makes infections harder to treat.
METHOD: DH5-alpha strains of Escherichia coli were grown in Luria-Bertani broth, inoculated in 24-well polystyrene plates containing coverslips, and incubated at 37C for one week. Biofilm disruption techniques were followed utilizing 0.05% vs 0.1% Tween-20, Triton X-100, and Sodium Dodecyl Sulfate (SDS). Crystal violet was used to stain the bacteria in the biofilms, and qualitative observations of the detergent’s impact were made under the microscope.
RESULTS: 0.1% concentrations of Tween-20 and SDS worked effectively at disrupting the biofilm than 0.05% concentrations. However, we observed that 0.05% concentrations of Triton X-100 worked better than 0.1% concentrations.
DISCUSSION/CONCLUSION: Future work will use fluorescent dyes, such as CFSE and Propidium iodide, to quantify the proportion of live vs dead bacteria to get more insight into the composition of bacteria in the biofilms. Analysis will also be done to evaluate the effectiveness of alternative anti-biofilm strategies, such as the use of extracellular enzymes and quorum quenching, and how to use them in tandem with antibiotic delivery agents to attack gram-negative bacteria.