In vitro and in silico analysis of antibiotic resistant in copper resistant bacteria klebsiella pneumoniae CJK 500 2.1.2

Ong, Michael Kayne (2024) In vitro and in silico analysis of antibiotic resistant in copper resistant bacteria klebsiella pneumoniae CJK 500 2.1.2. Bachelor thesis, Universitas Pelita Harapan.

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Abstract

The purposes of this study is to understand the antimicrobial resistance profiles of Klebsiella pneumoniae CJK 500 2.1.2 in copper as a candidate of bioremediation agent. In order to achieve the objective, K. pneumoniae CJK 500 2.1.2 is analyzed with antimicrobial testing by disc-diffusion, minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) method. Antimicrobial resistance gene, virulence factor, mobile elements, and genome characteristics of K. pneumoniae CJK 500 2.1.2 was evaluated using bioinformatic analysis. The bacterium was against 15 class of antibiotic in disc-diffusion testing and 2 class of antibiotics were chosen for MIC test. The MIC test used the microdilution method in 96-well plates. The MIC result was determined visually and using microplate reader. The MBC test conducted after the MIC test was done by inoculate 25 μl on the surface of a agar plate using swab method. All antimicrobial resistance testing use 4 mM of CuSO4 for copper treatment. The result shows that the bacterium able resist most of the antibiotic class with exceptions of sulphonamide and cefoxitin in the presence of copper. The MIC test result show that the bacterium is susceptible to tetracycline and cefoxitin with and without copper. The result indicates that there may no co-resistance/cross-resistance between antibiotic and heavy metal resistance. The MIC result may indicate interactions between antibiotic and copper. The bioinformatic analysis show that most of antimicrobial resistance mechanism of K. pneumoniae CJK 500 2.1.2 is efflux pumps. There are no significant result for virulence factors, and there are two insertion sequence presence in the bacterium chromosome. The antimicrobial genes (AMR) are not close to the mobile elements which could eliminate the possibility of horizontal gene transfer. The suggestions for future of this study are using additional antibiotic class for MIC method to achieve definite result. Also, if the bacterium will be used as bioremediation agent, the MIC and accumulation of copper study is recommended.

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