The antibiotic susceptibility profile of the isolates was identified using the standard disk diffusion method on Mueller–Hinton agar (bioMérieux, Marcy l’Etoile, France). Sixteen different antibiotics were tested, including amoxicillin (20 μg), amoxicillin-clavulanic acid (20/10 μg), piperacillin-tazobactam (30/6 μg), cephalotin (30 μg), ceftriaxone (30 μg), cefepime (30 μg), ertapenem (10 μg), imipenem (10 μg), amikacin (30 μg), gentamicin (10 μg), ciprofloxacin (5 μg), Fosfomycin (200 μg), nitrofurantoin (100 μg), tobramycin (10 μg), trimethoprim-sulfamethoxazole (1.25/23.75 μg), and colistin (10 μg) (bioMérieux, Marcy l’Etoile, France). The minimal inhibitory concentration (MIC) of colistin, ertapenem, and imipenem was identified using the microdilution and the E-test methods, respectively. Each strain was considered to be resistant to colistin, ertapenem, and imipenem if their MICs were greater than 2 mg/L, 1 mg/L, and 8 mg/L, respectively. The results were interpreted according to the European Microbial Medical Sensitivity Committee (EUCAST) 2017 (http://www.Sfmicrobiology.Org/Userfiles/Files/Files/CASFM/CASF%20V2_0_MAI2017.PDF , accessed on 25 September 2020).
Tobramycin
Manufactured by bioMérieux
Sourced in France
Tobramycin is a laboratory instrument used for the detection and quantification of the antibiotic tobramycin in biological samples. It is designed to provide accurate and reliable results for therapeutic drug monitoring and clinical research applications.
Automatically generated - may contain errors
Lab products found in correlation
Ciprofloxacin, by bioMérieux (4 mentions)
Imipenem, by bioMérieux (4 mentions)
Gentamicin, by bioMérieux (3 mentions)
Colistin, by bioMérieux (3 mentions)
Amikacin, by bioMérieux (3 mentions)
Cefepime, by bioMérieux (3 mentions)
Amoxicillin, by bioMérieux (2 mentions)
Amoxicillin clavulanic acid, by bioMérieux (2 mentions)
Ceftriaxone, by bioMérieux (2 mentions)
Nalidixic acid, by bioMérieux (2 mentions)
Piperacillin, by bioMérieux (2 mentions)
Ertapenem, by bioMérieux (1 mentions)
Nitrofurantoin, by bioMérieux (1 mentions)
Piperacillin tazobactam, by bioMérieux (1 mentions)
Fosfomycin, by bioMérieux (1 mentions)
Trimethoprim sulfamethoxazole, by bioMérieux (1 mentions)
Mueller hinton agar (mha), by bioMérieux (1 mentions)
Cefotaxime, by bioMérieux (1 mentions)
Kanamycin, by bioMérieux (1 mentions)
Ofloxacin, by bioMérieux (1 mentions)
6 protocols using tobramycin
1
Antibiotic Susceptibility Profiling of Isolates
2
Antibiotic Susceptibility Testing Protocol
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The antibiotics susceptibility pattern of the isolates was determined using the disk diffusion method on Mueller-Hinton agar (Oxoid, England). Inhibition zone diameter values were interpreted as recommended by the Antibiogram Committee of the French Society of Microbiology [11 ]. The 13 antibiotics (BioMérieux, France) used in this study are amoxicillin/clavulanic acid (20/10 μg), cefotaxime (30 μg), ceftriaxone (30 μg), amoxicillin (30 μg), imipenem (10 μg), gentamicin (10 μg), tobramycin (10 μg), amikacin (30 μg), kanamycin (30 μg), nalidixic acid (30 μg), ofloxacin (5 μg), ciprofloxacin (5 μg), and trimethoprim/sulfamethoxazole (1.25/23.75 μg).
3
Antimicrobial Susceptibility Profiling
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MICs were determined by the Etest method according to the manufacturer’s instructions. Antimicrobial agents included aztreonam, ceftazidime, ciprofloxacin, colistin, gentamicin, imipenem, nalidixic acid, tetracycline, tigecycline, tobramycin, and trimethoprim (bioMe’rieux, Marcy-l’_Etoile, France). E. coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853 were used as quality control strains. Interpretation of antimicrobial susceptibility was based on guidelines of the Clinical Laboratory Standards Institute (CLSI) [49 ].
4
Antimicrobial Potential of Boswellia integerrima Fractions
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Antimicrobial activities of total extract and fractions (Fr.1 - Fr.3) of B. integerrima were assessed using the standard minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Briefly, one row line in each plate was used for tobramycin (BioMerieux, Marcy-L’Etoile, France) as positive control in a serial dilution of 32-0.015 μg/mL.
The stock sample solutions were diluted and transferred into the next wells, and serial dilutions were made so that concentrations in the range of 2000, 1000, 500, 250, 125, 62.50, 31.25, 15.62, 7.81, 3.90, 1.95 and 0.97 μg/mL were obtained. Then 5 μL of the inoculum density equivalent to 0.5 McFarland of Brucella suspension was added to each well and incubated in shaker-incubator at 37 °C for 17 h, and the wells optical density (OD) were examined for turbidity, indicating growth of the bacteria. The lowest concentration of samples that inhibited bacterial growth (lack of visual turbidity), was regarded as the MIC. Then, 100 μL of cell suspensions of MIC and 4 next dilutions showing no bacterial growth were cultured again on Mueller-Hinton agar plates and incubated for 24 h. MBC was regarded as the first lowest concentration after MIC which showed no bacterial growth on plates. Each concentration was repeated three times to ensure reproducibility (13 (link)).
The stock sample solutions were diluted and transferred into the next wells, and serial dilutions were made so that concentrations in the range of 2000, 1000, 500, 250, 125, 62.50, 31.25, 15.62, 7.81, 3.90, 1.95 and 0.97 μg/mL were obtained. Then 5 μL of the inoculum density equivalent to 0.5 McFarland of Brucella suspension was added to each well and incubated in shaker-incubator at 37 °C for 17 h, and the wells optical density (OD) were examined for turbidity, indicating growth of the bacteria. The lowest concentration of samples that inhibited bacterial growth (lack of visual turbidity), was regarded as the MIC. Then, 100 μL of cell suspensions of MIC and 4 next dilutions showing no bacterial growth were cultured again on Mueller-Hinton agar plates and incubated for 24 h. MBC was regarded as the first lowest concentration after MIC which showed no bacterial growth on plates. Each concentration was repeated three times to ensure reproducibility (13 (link)).
5
Antibiotic Resistance Profiling of P. aeruginosa
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Following the recommendations of the European Committee on Antimicrobial Susceptibility Testing (EUCAST) (EUCAST 2019 ), both P. aeruginosa isolates were tested against the following antibiotic discs using the disc diffusion method. The multidrug discs contained ciprofloxacin (5 µg), piperacillin (100 µg), imipenem (10 µg), cefepime (30 µg), fosfomycin (200 µg), colistin (10 µg), and tobramycin (10 µg) (AB Biodisk, UK). The zones of inhibition (ZoI) were measured in millimetres following 12–18 h incubation at 37 °C, and interpreted in accordance with the manufacturer’s recommendations. Bacterial isolates were designated as antibiotic resistant (AMR) if they were resistant to multiple antimicrobial agents, classes, or subclasses of antibiotics as defined by EUCAST (Magiorakos et al. 2012 (link)).
6
Antimicrobial Susceptibility of Pseudomonas aeruginosa
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The isolated JNQH-PA57 was grown in Mueller-Hinton agar (MHA) (Oxoid, Hampshire, United Kingdom) at 37 °C for 24 h. The species identification was determined with Microflex LT/SH MALDI-TOF mass spectrometer (Bruker, Germany). The antimicrobial susceptibility of this stain was performed using MIC evaluation via the E-test method for the following antimicrobial agents: piperacillin, piperacillin/tazobactam, ticarcillin/clavulanic acid, ceftazidime, cefepime, aztreonam, imipenem, meropenem, amikacin, tobramycin, levofloxacin, ciprofloxacin, according to the manufacturer’s guidelines (AB Biodisk, Sweden). For colistin, the MIC was determined via a broth microdilution method, according to the Clinical and Laboratory Standards Institute (CLSI) guideline. P. aeruginosa ATCC 27853 served as a quality control strain for susceptibility testing. The interpretation of the results was based on the CLSI 2020 guideline [55 ].
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