Chonnam Med J.  2018 Jan;54(1):17-23. 10.4068/cmj.2018.54.1.17.

Salicylic Acid Reduces OmpF Expression, Rendering Salmonella enterica Serovar Typhimurium More Resistant to Cephalosporin Antibiotics

• Choi KM
• Kim MH
• Cai H
• Lee YJ
• Hong Y
• Ryu PY

• KMID: 2458580
• DOI: http://doi.org/10.4068/cmj.2018.54.1.17

Abstract

Salmonella enterica serovar Typhimurium is one of the most important bacterial pathogens causing diarrhea. The resistance of S. typhimurium to antimicrobial agents, which has recently been isolated from patients, is causing serious problems. We investigated the effects of salicylic acid (Sal) and acetyl salicylate (AcSal) on the susceptibility of S. typhimurium to cephalosporin antibiotics, which are known to increase resistance to cephalosporin and quinolone antibiotics. The MIC of cephalosporin antibiotics was higher than that of the media without Sal. The rate of accumulation of ethidium bromide (EtBr) in the bacteria by the outer membrane protein (Omp) was not different from that of the bacteria cultured in the medium containing Sal. However, Carbonyl cyanide-m-chlorophenylhydrazone (CCCP), an inhibitor of bacterial efflux pumps, significantly reduced the rate of accumulation of EtBr in bacteria cultured on Sal containing medium. In the medium containing CCCP, the MIC of the antimicrobial agent tended to decrease as compared with the control. In addition, the MIC of the bacteria treated with CCCP and Sal was higher than that of the antimicrobial agent against the CCCP treated experimental bacteria. These results suggest that Sal decreases the expression of OmpF in the Omp of S. typhimurium and reduces the permeability of cephalosporin antibiotics to bacteria, which may induce tolerance to cephalosporin antibiotics.

Keyword

Salmonella typhimurium; Salicylic Acid; Cephalosporin Resistance; OmpC protein; Anti-Bacterial Agents

MeSH Terms

Anti-Bacterial Agents
Anti-Infective Agents
Bacteria
Carbonyl Cyanide m-Chlorophenyl Hydrazone
Cephalosporin Resistance
Cephalosporins*
Diarrhea
Ethidium
Humans
Membrane Proteins
Permeability
Salicylic Acid*
Salmonella enterica*
Salmonella typhimurium
Salmonella*
Serogroup*
Anti-Bacterial Agents
Anti-Infective Agents
Carbonyl Cyanide m-Chlorophenyl Hydrazone
Cephalosporins
Ethidium
Membrane Proteins
Salicylic Acid

Figure

• 

  FIG. 1 Accumulation of EtBr in S. typhimurium by treated with Sal (5 mM) or AcSal (5 mM), and CCCP (100 µM) was added at the same time with ethidium bromide. EtBr accumulation in bacteria was measured as bacterial fluorescence intensity.

• 

  FIG. 2 Effect of Sal on EtBr accumulation in S. typhimurium. (A and C) EtBr accumulation in bacteria treated with 5 mM Sal (A) or AcSal (C). (B and D) EtBr accumulation in bacteria treated with indicated concentrations of Sal (B) or AcSal (D) in the presence of 100 µM CCCP.

• 

  FIG. 3 Effects of Sal on EtBr accumulation are increased by CCCP in S. typhimurium. Bacteria were left untreated (control) or treated with 5 mM Sal or AcSal. CCCP (100 µM) was added at the indicated time (arrow) and further cultured.

• 

  FIG. 4 The change in Omp protein levels by Sal in S. typhimurium. Omp proteins were isolated from bacteria cultured overnight in the absence (control) or presence (Sal) of 5 mM Sal. Same amounts of proteins were separated in SDS-PAGE and stained with Coomassie blue. M: protein standards.

• 

  FIG. 5 Effects of Sal on the expression of outer membrane protein genes and active efflux pump genes, quantified by real-time RT-PCR. Bacteria were cultured overnight in the absence or presence of 5 mM Sal and the amounts of the transcripts were measured using Q-RT-PCR analysis against their RNAs. The experiment was carried out triplicate.

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