The isolates were suspended and diluted in normal saline to 108 colony-forming units (CFU)/ml by comparison with a McFarland 0.5 turbidity standard, and spread onto Mueller–Hinton agar plates (Mast Diagnostics, Merseyside, UK) as recommended by the CLSI. The following disks (Mast Diagnostics) were tested: IMP (10 μg), IMP (10 μg) with APB (300 μg), meropenem (10 μg), meropenem (10 μg) with APB (300 μg), ceftazidime (30 μg), ceftazidime (30 μg) with APB (300 μg), IMP (10 μg), IMP (10 μg) with APB (600 μg), meropenem (10 μg), meropenem (10 μg) with APB (600 μg), ceftazidime (30 μg), and ceftazidime (30 μg) with APB (600 μg). APB (Sigma-Aldrich, St. Louis, MO, USA) was dissolved in water at 50 mg/ml, and 6 and 12 μl (for the 300 and 600 μg concentrations, respectively) was applied per disk. A 5-mm difference in zone diameter was used as a cutoff to identify resistant isolates [14 (link)].
Meropenem
Manufactured by Mast Diagnostics
Sourced in United Kingdom, Germany
Meropenem is a broad-spectrum antibiotic that belongs to the carbapenem class of medications. It is a sterile powder for injection and is used to treat a variety of bacterial infections.
Automatically generated - may contain errors
Lab products found in correlation
Amikacin, by Mast Diagnostics (4 mentions)
Gentamicin, by Mast Diagnostics (4 mentions)
Imipenem, by Mast Diagnostics (4 mentions)
Ciprofloxacin, by Mast Diagnostics (3 mentions)
Ceftazidime, by Mast Diagnostics (3 mentions)
Cefepime, by Mast Diagnostics (3 mentions)
Ceftriaxone, by Mast Diagnostics (3 mentions)
Aztreonam, by Mast Diagnostics (2 mentions)
Mueller hinton agar (mha), by Merck Group (2 mentions)
Ampicillin sulbactam, by Mast Diagnostics (2 mentions)
Cefotaxime, by Mast Diagnostics (2 mentions)
Trimethoprim sulfamethoxazole, by Mast Diagnostics (2 mentions)
Levofloxacin, by Mast Diagnostics (1 mentions)
Colistin, by Mast Diagnostics (1 mentions)
E test, by Liofilchem (1 mentions)
Meropenem, by Liofilchem (1 mentions)
Colistin, by Liofilchem (1 mentions)
Amikacin, by Merck Group (1 mentions)
Meropenem, by Thermo Fisher Scientific (1 mentions)
Nitrofurantoin, by Thermo Fisher Scientific (1 mentions)
7 protocols using meropenem
1
Antibiotic Resistance Profiling of Isolates
2
Antibiotic Susceptibility Testing of Klebsiella pneumoniae
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Using the Clinical and Laboratory Standards Institute (CLSI) guidelines [9 ], an antibiogram assay was performed on the isolated K. pneumoniae colonies. The antibiotic discs contained ampicillin (30 µg), cotrimoxazole (25 µg), cefixime (5 µg), cefotaxime (30 µg), ceftriaxone (30 µg), ceftazidime (30 µg), gentamicin (10 µg), amikacin (30 µg), tetracycline (30 µg), doxycycline (30 µg), minocycline (30 µg), tigecycline (15 µg), ciprofloxacin (5 µg), levofloxacin (5 µg), ampicillin/sulbactam (100/10 µg), piperacillin/tazobactam (100/10 µg), imipenem (10 µg), meropenem (10 µg), ertapenem (10 µg), doripenem (10 µg), aztreonam (30 µg), colistin (10 µg), and fosfomycin (200 µg) (Mast Diagnostics, United Kingdom) [4 , 10 ].
The minimum inhibitory concentration (MIC) of imipenem, meropenem, and colistin for isolates resistant to carbapenems was determined by using E-test (Liofilchem, Italy) according to the 2021 CLSI guidelines [9 ].
The minimum inhibitory concentration (MIC) of imipenem, meropenem, and colistin for isolates resistant to carbapenems was determined by using E-test (Liofilchem, Italy) according to the 2021 CLSI guidelines [9 ].
3
Antibiotic Susceptibility Testing of Isolates
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Susceptibility of isolates to various antibiotics was determined by Disk Diffusion Agar and Broth microdilution methods as recommended by the Clinical and Laboratory Standards Institute (CLSI) [18 ].
The following antibiotic disks (Mast Diagnostics- UK), were tested: Amikacin (AK), Gentamicin (GM), Meropenem (MEM), Imipenem (IMI), Ceftazidime (CAZ), Cefepime (CMP) and Polymixine B (PB). Escherichia coli ATCC 25922 was used as a control for susceptibility testing.
The MICs of Amikacin (Sigma Aldrich, St Louis, USA) and Ceftazidime (Jaber Ebne Hayyan Co, Iran) were determined by CLSI broth microdilution method (MIC range,0.5 to 256 μg/ml). P. aeruginosa ATCC 27853 were used as a control for quality assurance of the test.
The following antibiotic disks (Mast Diagnostics- UK), were tested: Amikacin (AK), Gentamicin (GM), Meropenem (MEM), Imipenem (IMI), Ceftazidime (CAZ), Cefepime (CMP) and Polymixine B (PB). Escherichia coli ATCC 25922 was used as a control for susceptibility testing.
The MICs of Amikacin (Sigma Aldrich, St Louis, USA) and Ceftazidime (Jaber Ebne Hayyan Co, Iran) were determined by CLSI broth microdilution method (MIC range,0.5 to 256 μg/ml). P. aeruginosa ATCC 27853 were used as a control for quality assurance of the test.
4
Antimicrobial Susceptibility Testing of A. baumannii
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Antimicrobial susceptibility testing was performed on Mueller-Hinton agar (MHA) (Merck, Co., Germany) by agar disk diffusion (DD) method as recommended by the Clinical and Laboratory Standards Institute (CLSI document M100-S14) [17 ]. The tested antibiotics were as follows: amikacin (AK; 30 μg), ciprofloxacin (CP; 5 μg), ceftazidime (CAZ; 30 μg), gentamicin (GM; 10 μg), imipenem (IMP; 10 μg), meropenem (MER; 10 μg) and piperacillin/tazobactam (PTZ; 100/10 μg) (MAST diagnostics, Merseyside, UK). A. baumannii ATCC 17978 was used as a positive quality control (PQC) and P. aeruginosa ATCC 25853 and E. coli ATCC 25922 were used as a negative quality control (NQC) in this study.
5
Antibiotic Susceptibility of E. coli
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Initially, all Isolates were tested using the Kirby Bauer disk diffusion method according to the Clinical Laboratory Standards Institute (2013) . E. coli ATCC25922 was used as control strains. The tested antibiotics included: ampicillin, ampicillin/sulbactam, amoxicillin/clavulanate, ticarcillin, piperacillin, piperacillin/tazobactam, cephalothin, cefuroxime, cefoxitin, ceftriaxone, cefotaxime, ceftazidime, cefepime, meropenem, and aztreonam (Mast Diagnostics, Merseyside, UK). co-trimoxazole, gentamicin, nalidixic acid, ciprofloxacin, imipenem, meropenem, tetracycline, and nitrofurantoin (Oxoid Ltd., Basingstoke, UK). MIC values were extrapolated by the BIOMIC automated reading system and software package (Giles Scientific, New York, NY, USA).
The disk diffusion method was applied to assess ESBL production in all the Isolates with cefotaxime (30 ug) and ceftazidime (30 ug) alone and in combination with clavulanic acid (10 ug) as recommended by the Clinical and Laboratory Standards Institute (CLSI).
The disk diffusion method was applied to assess ESBL production in all the Isolates with cefotaxime (30 ug) and ceftazidime (30 ug) alone and in combination with clavulanic acid (10 ug) as recommended by the Clinical and Laboratory Standards Institute (CLSI).
6
Antibiotic Susceptibility Testing of Bacterial Isolates
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According to the clinical and laboratory standards Institute protocol (CLSI; M100-S14) [13 ], the antibiotic susceptibility was tested by disk agar diffusion method on the Mueller-Hinton agar plates (MHA) (Merck, Darmstadt, Germany) for ceftazidime (CAZ: 30 µg), cefotaxime (CTX: 30 µg), imipenem (IPM: 10 µg), meropenem (MEM: 10 µg), ciprofloxacin (CIP: 5 µg), cefepime (FEP: 30 µg), ceftriaxone (CRO: 30 µg), amikacin (AN: 30 µg), gentamicin (GM; 10 µg), and trimethoprim-sulfamethoxazole (SXT; 5 µg) (MAST Diagnostics, Merseyside, UK).
Strains non-susceptible to at least three or more antimicrobial classes were defined as MDR, and those non-susceptible to at least one agent in all but two or more antimicrobial categories were considered as possible XDR, and the strains that non-susceptibility to all agents in all antimicrobial categories were defined as possible pan drug-resistant (PDR) [14 (link)]. Escherichia coli ATCC 25922 was used as control organism in susceptibility testing.
Strains non-susceptible to at least three or more antimicrobial classes were defined as MDR, and those non-susceptible to at least one agent in all but two or more antimicrobial categories were considered as possible XDR, and the strains that non-susceptibility to all agents in all antimicrobial categories were defined as possible pan drug-resistant (PDR) [14 (link)]. Escherichia coli ATCC 25922 was used as control organism in susceptibility testing.
7
Antimicrobial Susceptibility Testing Protocol
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In concordance with the Clinical and Laboratory Standards Institute; CLSI. 2018 [16], antimicrobial susceptibility testing was done on the Mueller-Hinton agar plates (Merck Co., Germany) by disk agar diffusion (DD) method against 16 following antimicrobials: levo oxacin (LEV; 5 µg); ceftazidime (CAZ; 30 µg), cefotaxime (CTX; 30 µg ), cefepime (FEP; 30 µg), ertapenem (ETP; 10 µg), amikacin (AK; 30 µg), meropenem (MER; 10 µg), ceftriaxone (CRO; 30 μg), ampicillin/sulbactam (SAM; 10/10 µg), cefoperazone (CFP; 75 µg), imipenem (IPM; 10 µg), nitrofurantoin (NIT;300 µg), gentamicin (GM; 10 µg), cipro oxacin (CIP; 5 µg), tetracycline (TET; 30 µg), and trimethoprim-sulfamethoxazole (SXT; 5 µg) (MAST Diagnostics, Merseyside, UK). The MDR and possible XDR/PDR strains were recognized according to the guidelines suggested by the European Center for Disease Control and Prevention (ECDC) (17) . E. coli ATCC 25922 was used as a quality control (QC) organism. Also, colistin susceptibility assay was performed for carbapenems-resistance isolates by broth microdilution method according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints. The E. coli NCTC 13846 (a colistin resistant strain) was used as QC in colistin minimum inhibitory concentration (MIC) determination.
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