Complete Proteome of a Quinolone-Resistant Salmonella Typhimurium Phage Type DT104B Clinical Strain

Abstract

Salmonellosis is one of the most common and widely distributed foodborne diseases. The emergence of Salmonella strains that are resistant to a variety of antimicrobials is a serious global public health concern. Salmonella enterica serovar Typhimurium definitive phage type 104 (DT104) is one of these emerging epidemic multidrug resistant strains. Here we collate information from the diverse and comprehensive range of experiments on Salmonella proteomes that have been published. We then present a new study of the proteome of the quinolone-resistant Se20 strain (phage type DT104B), recovered after ciprofloxacin treatment and compared it to the proteome of reference strain SL1344. A total of 186 and 219 protein spots were recovered from Se20 and SL1344 protein extracts, respectively, after two-dimensional gel electrophoresis. The signatures of 94% of the protein spots were successfully identified through matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS). Three antimicrobial resistance related proteins, whose genes were previously detected by polymerase chain reaction (PCR), were identified in the clinical strain. The presence of these proteins, dihydropteroate synthase type-2 (sul2 gene), aminoglycoside resistance protein A (strA gene) and aminoglycoside 6'-N-acetyltransferase type Ib-cr4 (aac(6')-Ib-cr4 gene), was confirmed in the DT104B clinical strain. The aac(6')-Ib-cr4 gene is responsible for plasmid-mediated aminoglycoside and quinolone resistance. This is a preliminary analysis of the proteome of these two S. Typhimurium strains and further work is being developed to better understand how antimicrobial resistance is developing in this pathogen.

Planning a One Health Case Study to Evaluate Methicillin Resistant Staphylococcus aureus and Its Economic Burden in Portugal

“…Beyond surveillance studies, our research group has investigated AMR mechanisms by characterizing a range of resistant strains of interest through proteomic approaches in MRSA (Monteiro et al, 2012, 2015) and other bacterial species (Pinto et al, 2010; Radhouani et al, 2010, 2012; Correia et al, 2014, 2016; Goncalves et al, 2014; Ramos et al, 2015, 2016; Monteiro et al, 2016). However, these proteomic studies, and most AMR research in general, have focused on bacteria growing in planktonic cultures and hence overlooked biofilm-specific AMR mechanisms.…”

Current perspectives on the dynamics of antibiotic resistance in different reservoirs

“…Likewise, a complete proteome might be obtained through matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS). Correia et al (2014) reported confirmation of several proteins in S. Typhimurium phage type 104 clinical strain through this method, emphasizing the presence of the Aac(6 0 )-Ib-cr enzyme responsible for both plasmid-mediated aminoglycoside and quinolone resistance [37,57].…”

Applied Proteomics in ‘One Health’

“…The proteome of a fluoroquinolone-resistant S. enterica serovar Typhimurium Se20 isolate (phage type DT104B), recovered after ciprofloxacin treatment, was studied compared to the proteome of a susceptible reference isolate SL1344. In the isolate recovered after antibiotic administration, three proteins related to antimicrobial resistance were detected [ 234 ]. Subsequently, ciprofloxacin-resistant and -susceptible isolates of Salmonella were used and tandem mass tag labelling and acetylation enrichment techniques were employed to screen for the different expression of acetylated proteins between the two isolates and for quantitative and bioinformatics analysis.…”