Team:YAU-China/Results
Results
In order to verify whether the engineered bacteria we constructed can effectively destroy the Pseudomonas aeruginosa biofilm, we designed the following experiments:
1: Engineered bacteria on the destruction of wild bacteria biofilm
The destruction of the formed Pseudomonas aeruginosa PAO1 biofilm by engineered bacteria: Figure A is the image of biofilm cultured by shear force under laser confocal, excited by argon laser (488nm) and photographed under objective lens×10; Figure B is the crystal violet staining OD570 of the biofilm deposited in the PVC plate by 95% ethanol.
2: Engineered bacteria against wild bacteria biofilm prevention experiment
Co-culture of engineered bacteria with Pseudomonas aeruginosa PAO1 to form biofilm: Figure C is the image of biofilm cultured by shear force under laser confocal, excited by argon laser (488nm) and photographed under objective lens×10; Figure D shows the crystal violet staining OD570 of the biofilm deposited in a PVC plate by 95% ethanol.
3: Cracking efficiency of engineered bacteria
Figure E shows the bacterial survival counts of engineered and wild bacteria at different times when the engineered bacteria and Pseudomonas aeruginosa were co-cultured.
4: Antibiotic susceptibility test
Counting the survival of Pseudomonas aeruginosa bacteria in the biofilm of Pseudomonas aeruginosa treated with tobramycin and engineering bacteria.
(1) Drug sensitivity test after biofilm destruction
(2) Co-culture biofilm susceptibility test
Figure F shows the number of bacteria surviving in the biofilm after adding engineering bacteria and tobramycin treatment to the biofilm of Pseudomonas aeruginosa wild bacteria in static culture. Figure G shows the number of bacteria surviving in the biofilm after treatment with tobramycin added to the biofilm formed by co-culture of Pseudomonas aeruginosa and engineered bacteria.
The engineered bacteria constructed by our team can effectively destroy the biofilm of Pseudomonas aeruginosa, whether it is a biofilm formed by static growth or sheared growth; and after treatment with engineering bacteria, tobramycin can kill Pseudomonas encapsulated in biofilm.
We believe that it is effective to apply engineered bacteria to clinically destroy biofilms, and that the engineered bacteria have a small number of bacterial survival after the biofilm is processed due to the lysis mechanism, and other safety mechanisms designed by the experiment can effectively prevent the escape of engineered bacteria, Therefore, greatly reduce the risk of our products being used in clinical treatment in the future.