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      Prevalence of Extended-Spectrum and Metallo β-Lactamase Production in AmpC β-Lactamase Producing Pseudomonas aeruginosa Isolates From Burns

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          Abstract

          Background:

          Pseudomonas aeruginosa is one of the most common causes of nosocomial infections. Resistance of P. aeruginosa to β-lactam antibiotics may be the result of acquired resistance through mutation and over production of various antibiotic inactivating enzymes. This research aimed to determine the prevalence of extended-spectrum β-lactamases (ESBL) and metallo β-lactamase (MBL) production as well as the presence of their related genes among AmpC β-lactamase producing P. aeruginosa isolated from burns.

          Objectives:

          The current study aimed to determine the prevalence of class A ESBL and MBL production in relation to the presence of their related genes among AmpC β-lactamase producing P. aeruginosa isolated from burns.

          Materials and Methods:

          The antimicrobial susceptibility of 51 P. aeruginosa isolates from patients with burns was examined against 13 antibiotics by the disc diffusion method. Minimum inhibitory concentrations (MIC) for imipenem and ceftazidime were measured by the microdilution method. AmpC production was detected by AmpC disc and the modified three-dimensional extract tests. ESBL phenotype was confirmed by the double disc synergy test (DDST). Presence of β-lactamase genes was detected by specific primers and polymerase chain reaction (PCR).

          Results:

          All isolates were multidrug resistant. AmpC, ESBL and MBL production were observed in 35 (68.6%), 20 (39.2%) and 19 (37.3%) isolates, respectively. Overall, 43 isolates (84.3%) carried β-lactamase genes, out of which 31 (60.8%) harbored bla AmpC , 20 (39.2%) had bla TEM and 11 (21.6%) carried bla PER -1 genes. Among the AmpC producers, two isolates (6.5%) carried bla AmpC + bla ESBL , 13 (41.9%) had bla AmpC + bla MBL and six (19.4%) produced the three enzymes.

          Conclusions:

          A high prevalence of multiple β-lactamase production was observed among the AmpC producers (60%), of which the majority co-produced AmpC and MBL. The current study results showed correlation between β-lactamase production and the presence of antibiotic resistance genes in the isolates.

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          Most cited references31

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          Extended-Spectrum β-Lactamases: a Clinical Update

          Extended-spectrum β-lactamases (ESBLs) are a rapidly evolving group of β-lactamases which share the ability to hydrolyze third-generation cephalosporins and aztreonam yet are inhibited by clavulanic acid. Typically, they derive from genes for TEM-1, TEM-2, or SHV-1 by mutations that alter the amino acid configuration around the active site of these β-lactamases. This extends the spectrum of β-lactam antibiotics susceptible to hydrolysis by these enzymes. An increasing number of ESBLs not of TEM or SHV lineage have recently been described. The presence of ESBLs carries tremendous clinical significance. The ESBLs are frequently plasmid encoded. Plasmids responsible for ESBL production frequently carry genes encoding resistance to other drug classes (for example, aminoglycosides). Therefore, antibiotic options in the treatment of ESBL-producing organisms are extremely limited. Carbapenems are the treatment of choice for serious infections due to ESBL-producing organisms, yet carbapenem-resistant isolates have recently been reported. ESBL-producing organisms may appear susceptible to some extended-spectrum cephalosporins. However, treatment with such antibiotics has been associated with high failure rates. There is substantial debate as to the optimal method to prevent this occurrence. It has been proposed that cephalosporin breakpoints for the Enterobacteriaceae should be altered so that the need for ESBL detection would be obviated. At present, however, organizations such as the Clinical and Laboratory Standards Institute (formerly the National Committee for Clinical Laboratory Standards) provide guidelines for the detection of ESBLs in klebsiellae and Escherichia coli . In common to all ESBL detection methods is the general principle that the activity of extended-spectrum cephalosporins against ESBL-producing organisms will be enhanced by the presence of clavulanic acid. ESBLs represent an impressive example of the ability of gram-negative bacteria to develop new antibiotic resistance mechanisms in the face of the introduction of new antimicrobial agents.
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            Multiple mechanisms of antimicrobial resistance in Pseudomonas aeruginosa: our worst nightmare?

            Pseudomonas aeruginosa carries multiresistance plasmids less often than does Klebsiella pneumoniae, develops mutational resistance to cephalosporins less readily than Enterobacter species, and has less inherent resistance than Stenotrophomonas maltophilia. What nevertheless makes P. aeruginosa uniquely problematic is a combination of the following: the species' inherent resistance to many drug classes; its ability to acquire resistance, via mutations, to all relevant treatments; its high and increasing rates of resistance locally; and its frequent role in serious infections. A few isolates of P. aeruginosa are resistant to all reliable antibiotics, and this problem seems likely to grow with the emergence of integrins that carry gene cassettes encoding both carbapenemases and amikacin acetyltransferases.
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              Extended-spectrum cephalosporinases in Pseudomonas aeruginosa.

              The characterization of AmpC-type beta-lactamases was performed in a collection of 32 clinical Pseudomonas aeruginosa isolates with intermediate susceptibility or resistance to imipenem and ceftazidime. Twenty-one out of those 32 isolates overexpressed AmpC beta-lactamase, and the MICs of ceftazidime and imipenem were reduced after cloxacillin addition. Cloning and sequencing identified 10 AmpC beta-lactamase variants. Reduced susceptibility to imipenem, ceftazidime, and cefepime was observed only with recombinant P. aeruginosa strains expressing an AmpC beta-lactamase that had an alanine residue at position 105. The catalytic efficiencies (k(cat)/K(m)) of the AmpC variants possessing this residue were increased against oxyiminocephalosporins and imipenem. In addition, we show here that those AmpC variants constitute a favorable background for the in vitro selection of imipenem-resistant strains. This report identified a novel resistance mechanism that may contribute to imipenem resistance in P. aeruginosa.
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                Author and article information

                Journal
                Jundishapur J Microbiol
                Jundishapur J Microbiol
                10.5812/jjm
                Kowsar
                Jundishapur Journal of Microbiology
                Kowsar
                2008-3645
                2008-4161
                September 2014
                23 September 2014
                : 7
                : 9
                : e16436
                Affiliations
                [1 ]Department of Microbiology, Faculty of Biological Sciences, Shahid Beheshti University, Tehran, IR Iran
                [2 ]Department of Pediatrics, Shahid Beheshti University of Medical Sciences, Tehran, IR Iran
                Author notes
                [* ]Corresponding author: Fereshteh Eftekhar, Department of Microbiology, Faculty of Biological Sciences, Shahid Beheshti University, Tehran, IR Iran. Tel: +98-2129903208, Fax: +98-2122431664, E-mail: f-eftekhar@ 123456sbu.ac.ir
                Article
                10.5812/jjm.16436
                4255381
                25485066
                3b0e6f5c-1118-40fe-b888-220b07755ca2
                Copyright © 2014, Ahvaz Jundishapur University of Medical Sciences; Published by Kowsar.

                This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License ( http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.

                History
                : 15 December 2013
                : 09 June 2014
                : 21 June 2014
                Categories
                Research Article

                pseudomonas aeruginosa,beta-lactamase,blaampc,blaesbl,blambl

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