Evaluating the Effectiveness of Chemical Agents and OMVs in Disrupting Biofilm in Escherichia coli
Faculty Mentor
Vinaya Sampath
Area of Research
Microbiology
Major
Forensic Science and Biology
Description
Gram-negative bacteria have been a major obstacle in the field of medicine, due to its resistance against antibiotics. This resistance is due to the production of biofilm, which is an extracellular matrix that surrounds the bacteria and prevents antibiotics from passing through. In order to begin tackling this issue, DH5-alpha strains of Escherichia coli were grown in LB, inoculated in 96-well polystyrene plates, and left to rest at 37 degrees Celsius for one week to help with biofilm growth. Varying concentrations of detergents such as Tween-20, Triton X-100, and Sodium Dodecyl Sulfate (SDS), along with OMVs produced by Pseudomonas aeruginosa, were then tested as factors to disrupt or displace biofilm produced by E. coli. Biofilm displacement called for the detergents and OMVs to be inoculated in 96-well polystyrene plates an hour after the biofilm developed for a week, while biofilm disruption called for the detergents and OMVs to be inoculated alongside the E. coli under the above-mentioned conditions. Biofilm growth was measured based on the amount of crystal violet staining on the wells of the plates, which was then normalized to optical density measurements taken at 600 nm to determine how effective the detergents and OMVs were at impacting the biofilm. Once these observations have been made, I would like to look into the possibility of using OMVs as delivery agents to directly present antibiotics to bacteria such as P.aeruginosa. I would like to further look into anti-biofilm factors tested in previous studies, such as the use of extracellular enzymes, quorum quenching, and combining antibiotics with factors such as antimicrobial peptides or amino acids, to test whether these are more effective than the above-mentioned strategies.
Evaluating the Effectiveness of Chemical Agents and OMVs in Disrupting Biofilm in Escherichia coli
Gram-negative bacteria have been a major obstacle in the field of medicine, due to its resistance against antibiotics. This resistance is due to the production of biofilm, which is an extracellular matrix that surrounds the bacteria and prevents antibiotics from passing through. In order to begin tackling this issue, DH5-alpha strains of Escherichia coli were grown in LB, inoculated in 96-well polystyrene plates, and left to rest at 37 degrees Celsius for one week to help with biofilm growth. Varying concentrations of detergents such as Tween-20, Triton X-100, and Sodium Dodecyl Sulfate (SDS), along with OMVs produced by Pseudomonas aeruginosa, were then tested as factors to disrupt or displace biofilm produced by E. coli. Biofilm displacement called for the detergents and OMVs to be inoculated in 96-well polystyrene plates an hour after the biofilm developed for a week, while biofilm disruption called for the detergents and OMVs to be inoculated alongside the E. coli under the above-mentioned conditions. Biofilm growth was measured based on the amount of crystal violet staining on the wells of the plates, which was then normalized to optical density measurements taken at 600 nm to determine how effective the detergents and OMVs were at impacting the biofilm. Once these observations have been made, I would like to look into the possibility of using OMVs as delivery agents to directly present antibiotics to bacteria such as P.aeruginosa. I would like to further look into anti-biofilm factors tested in previous studies, such as the use of extracellular enzymes, quorum quenching, and combining antibiotics with factors such as antimicrobial peptides or amino acids, to test whether these are more effective than the above-mentioned strategies.