Role of efflux pumps: MexAB-OprM and MexXY(-OprA), AmpC cephalosporinase and OprD porin in non-metallo-β-lactamase producing Pseudomonas aeruginosa isolated from cystic fibrosis and burn patients.
OBJECTIVE
In order to gain a better understanding of the role of several mechanisms in antibiotic resistance in Pseudomonas aeruginosa clinical isolates obtained from CF and burn patients, we evaluated gene expression of efflux pumps MexAB-OprM and MexXY(-OprA), the natural β-lactamase AmpC and outer membrane porin protein OprD. Also, the presence of genes encoding Ambler classes A, B β-lactamases and aminoglycoside modifying enzymes (AMEs) was examined.


RESULTS
Piperacillin-tazobactam and amikacin retained the highest in vitro activities among 21 CF and 27 burn P. aeruginosa isolates. Based on Enterobacterial Repetitive Intergenic Consensus (ERIC) PCR, 15 distinct patterns were detected. There were 5 CF and 6 burn isolates harbored PER-1 and VEB-1, respectively. Among AMEs, involved in resistance of anti-Pseudomonas aminoglycosides, aac(6')-Ib was the most prevalent gene. Among CF isolates, mexA overexpression was the most prevalent mechanism (47.6%) followed by mexX (42.8%), ampC (9.5%) and oprD downregulation (4.7%). Among burn isolates, the prevalence of mexX, mexA, and ampC overexpression was 62.9%, 74%, and 11.1%, respectively. Downregulation of oprD was observed in 14.8% of burn isolates.


CONCLUSIONS
Among CF isolates, mexX and mexA overexpression were the major contributing factors to aminoglycoside (gentamicin) and carbapenem (meropenem) resistance, respectively while among burn isolates, AMEs in conjunction with mexX hyperexpression were identified to be responsible for aminoglycoside resistance. Also mexA overexpression was partially associated with carbapenem resistance. Moreover, cephalosporin resistance was linked to overexpression of mexA and/or mexX. The impact of interplay between different resistance mechanisms on resistant phenotypes was more complicated among burn than CF isolates.
Tabriz Research Center of Infectious and Tropical Diseases, Tabriz University of Medical Sciences, Tabriz, Iran.
Mohammad
M
Aghazadeh
Tabriz Research Center of Infectious and Tropical Diseases, Tabriz University of Medical Sciences, Tabriz, Iran; Microbiology Department, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran. Electronic address: hojabriz@yahoo.com.
Zoya
Z
Hojabri
Molecular Medicine Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
Reza
R
Mahdian
Microbiology Department, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
Mohammad Reza
MR
Nahaei
Department of Clinical Biochemistry and Laboratory, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
Marveh
M
Rahmati
Microbiology Department, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
Tina
T
Hojabri
Microbiology Department, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
Tahereh
T
Pirzadeh
Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran.
Omid
O
Pajand
D000583:Amikacin / Q000627:therapeutic use; D000900:Anti-Bacterial Agents; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis; D001426:Bacterial Proteins / Q000096:biosynthesis; D002056:Burns / Q000382:microbiology*; D002510:Cephalosporinase; D003550:Cystic Fibrosis / Q000382:microbiology*; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D026901:Membrane Transport Proteins / Q000096:biosynthesis; D008826:Microbial Sensitivity Tests; D010397:Penicillanic Acid / Q000031:analogs & derivatives / Q000627:therapeutic use; D010878:Piperacillin / Q000627:therapeutic use; D018272:Porins / Q000096:biosynthesis; D011552:Pseudomonas Infections / Q000188:drug therapy; D011550:Pseudomonas aeruginosa / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000078142:Tazobactam; D001618:beta-Lactamases / Q000096:biosynthesis
D000900:Anti-Bacterial Agents; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; C118598:MexA protein, Pseudomonas aeruginosa; C118597:MexB protein, Pseudomonas aeruginosa; C402791:MexXY protein, Pseudomonas aeruginosa; C402263:OprM protein, Pseudomonas aeruginosa; D018272:Porins; C077157:OprD protein, Pseudomonas aeruginosa; D000583:Amikacin; D010397:Penicillanic Acid; D002510:Cephalosporinase; C108863:beta-lactamase PER-1; C119424:beta-lactamase VEB-1, E coli; C048795:AmpC beta-lactamases; D001618:beta-Lactamases; D000078142:Tazobactam; D010878:Piperacillin
AmpC; Efflux pumps; Iran; OprD; Pseudomonas aeruginosa