Antibiotic susceptibility testing was determined on Müller–Hinton agar by standard disc diffusion method as recommended by the European Committee on Antimicrobial Susceptibility Testing (EUCAST; www.eucast.org ). Seventeen antibiotics were tested, including ticarcillin, ticarcillin-clavulanic acid, piperacillin, piperacillin-tazobactam, ceftazidime, cefotaxime, cefepime, aztreonam, amikacin, tobramycin, gentamicin, ciprofloxacin, rifampicin, ertapenem, meropenem, imipenem and colistin (Bio-Rad, Marnes-la-Coquette, France). The MIC for imipenem was determined using the Etest method (bioMérieux, La Balmes les Grottes, France) and the result was interpreted according to the EUCAST breakpoint for Enterobacteriaceae (susceptible if MIC ≤ 2 mg/L)
Tobramycin
Manufactured by Bio-Rad
Sourced in France
Tobramycin is a laboratory product manufactured by Bio-Rad. It is an antibiotic used for the detection and quantification of the drug Tobramycin in various biological samples.
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Lab products found in correlation
Ciprofloxacin, by Bio-Rad (9 mentions)
Cefotaxime, by Bio-Rad (7 mentions)
Imipenem, by Bio-Rad (7 mentions)
Gentamicin, by Bio-Rad (6 mentions)
Aztreonam, by Bio-Rad (6 mentions)
Piperacillin, by Bio-Rad (6 mentions)
Tetracycline, by Bio-Rad (6 mentions)
Chloramphenicol, by Bio-Rad (6 mentions)
Amikacin, by Bio-Rad (5 mentions)
Piperacillin tazobactam, by Bio-Rad (5 mentions)
Nalidixic acid, by Bio-Rad (5 mentions)
Ceftazidime, by Bio-Rad (4 mentions)
Cefepime, by Bio-Rad (4 mentions)
Cefoxitin, by Bio-Rad (4 mentions)
Netilmicin, by Bio-Rad (4 mentions)
Amoxicillin clavulanic acid, by Bio-Rad (4 mentions)
Trimethoprim sulfamethoxazole, by Bio-Rad (3 mentions)
Amoxicillin, by Bio-Rad (3 mentions)
Ceftriaxone, by Bio-Rad (3 mentions)
Cefixime, by Bio-Rad (3 mentions)
9 protocols using tobramycin
1
Evaluating Antibiotic Susceptibility with EUCAST Standards
2
Antimicrobial Susceptibility Testing Protocol
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Antimicrobial susceptibility testing was performed on all isolates recovered from the two selective media using the disc diffusion method on Mueller–Hinton (MH) agar plates (Neogen, Lansing, Michigan) for ticarcillin (75 μg), amoxicillin/clavulanic acid (20–10 μg), cefotaxime (30 μg), ceftazidime (10 μg), temocillin (30 μg), cefoxitin (30 μg), ertapenem (10 μg), imipenem (10 μg), meropenem (10 μg), ceftazidime/avibactam (10–4 μg), aztreonam (30 μg), ciprofloxacin (5 μg), trimethoprim-sulfamethoxazole (SXT) (1.25–23.75 μg), tetracycline (30 μg), amikacin (30 μg), gentamicin (15 μg), and tobramycin (10 μg) (Bio-Rad Laboratories, Algés, Portugal), following EUCAST recommendations and breakpoint tables. Susceptibility to fosfomycin was evaluated by the disk diffusion method (50 μg) on MH agar plates supplemented with 25 μg/mL glucose-6-phosphate, according to EUCAST guidelines [24 ]. Strain E. coli ATCC 25922 was used for quality control. Multidrug resistance was defined as acquired non-susceptibility to at least one agent in three or more antimicrobial categories [25 (link)].
3
Antimicrobial Susceptibility Profiling of MRSA
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Susceptibility to cefoxitin, chloramphenicol, ciprofloxacin, clindamycin, fusidic acid, erythromycin, gentamicin, kanamycin, linezolid, mupirocin, quinupristin-dalfopristin, penicillin, rifampin, tetracycline, tobramycin and trimethoprim/sulfamethoxazole (BioRad, USA) was performed by disk diffusion method and to vancomycin and teicoplanic by Etest (bioMérieux, France) in accordance to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) recommendation (http://www.eucast.org ). Multidrug resistance (MDR) was defined as resistance of MRSA to three or more district antimicrobial classes in addition to beta-lactams.
4
Antibiotic Susceptibility Testing by Kirby-Bauer
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The antibiotic susceptibility testing was performed using the Kirby–Bauer disc diffusion method for 16 antibiotics, namely ampicillin (10 µg), amoxicillin-clavulanic acid (30 µg), amikacin (30 µg), ceftazidime (30 µg), ciprofloxacin (5 µg), cefotaxime (30 µg), cefuroxime (30 µg), cefepime (30 µg), cefoxitin (30 µg), gentamicin (10 µg), meropenem (10 µg), moxifloxacin (5 µg), piperacillin (100 µg), trimethoprim-sulfamethoxazole (1.25 + 23.75 µg), tobramycin (10 µg), and piperacillin-tazobactam (10 µg) (Bio-Rad, Feldkirchen, Germany), and read using the ADAGIO 93400 automated system (Bio-Rad, Feldkirchen, Germany). The readings were interpreted as resistant, intermediate (susceptible with increased exposure), or susceptible according to the respective breakpoints for every organism in the European Committee on Antimicrobial Susceptibility Testing [42 ].
5
Antimicrobial Resistance Profiling of Diarrheagenic E. coli
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Thirty-one (31) diarrheagenic Eschericia coli (DEC) were subjected to the antimicrobial susceptibility testing. It was carried out by disk diffusion method on Müller-Hinton agar (Liofilchem, Italy) according to the recommendations of the European Committee on Antimicrobial Susceptibility Testing (EUCAST) [10 ]. After depositing the antibiotics, the plates were incubated at +37 °C for 18–24 h. The diameters of the antibiotic sensitivity halos were recorded according to the recommendations of EUCAST. Intermediary (I) susceptibility of pathovars was classified as resistant (R). According to EUCAST recommendations on antibiotics used in case of infection with enterobacteria and in view of the multidrug resistance observed in recent years, 19 antibiotics divided into 7 different families were tested. These include amoxicillin (25 µg), amoxicillin–clavulanic acid (20/10 µg), ceftriaxone (30 µg), cefotaxime (30 µg), cefepime (30 – g), cefixime (10 µg), piperacillin (75 µg), piperacillin–tazobactam (100 +10 µg), imipenem (10 µg), tetracycline (30 µg), chloramphenicol (30 µg), trimethoprim–sulfametoxazole (1.25 ± 23.75 µg), aztreonam (30 µg), colistin sulfate (50 µg), ciprofloxacin (5 µg), nalidixic acid (30 µg), gentamycin (15 µg), netilmicin (10 µg), and tobramycin (10 µg) (Bio-Rad, France).
6
Antimicrobial Susceptibility Testing Protocol
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Antimicrobial susceptibility testing was performed according to the 2016 recommendations of the Antibiogram Committee of the French Society for Microbiology1 . Disk diffusion method was performed for amikacin, amoxicillin +/- clavulanic acid, aztreonam, cefalotin, cefepime, cefoxitin, cefpirom, cefotaxime, ceftazidime, chloramphenicol, ciprofloxacin, colistin, cotrimoxazole, fosfomycin, gentamicin, imipenem, isepamicin, levofloxacin, moxalactam, nalidixic acid, netilmicin, ofloxacin, piperacillin, piperacillin/tazobactam, tetracycline, ticarcillin +/- clavulanic acid, and tobramycin (Bio-Rad®). E-test (bioMérieux®) was performed when inhibition zone diameters were observed around disks, and minimal inhibitory concentration (MIC) results that differed by more than two dilutions were considered as different. For ciprofloxacin, MICs were determined by the agar dilution reference method.
7
Antibiotic Susceptibility Profiling of Pathogens
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Antibiotic susceptibility was determined on Mueller-Hinton agar using the standard disc diffusion procedure as described by the European Committee of Antimicrobial Susceptibility Testing (EUCAST) [19] . Nineteen antibiotics belonging to 7 different families were tested : amoxicillin (25 µg), amoxicillin-clavulanic acid (20/10 µg), ceftriaxone (30 µg), cefotaxime (30 µg), cefepime (30 µg), cefixime (10 µg), piperacillin (75 µg), piperacillin-tazobactam (100 + 10 µg), imipenem (10 µg), tetracycline (30 µg), chloramphenicol (30 µg), trimethoprim-sulfametoxazole (1.25 ± 23.75 µg), aztreonam (30 µg), colistin sulfate (50 µg), ciprofloxacin (5 µg), nalidixic acid (30 µg), gentamycin (15 µg), netilmicin (10 µg) and tobramycin (10 µg) (Bio-Rad, France). The diameters of the antibiotic sensitivity halos were recorded according to the recommendations of EUCAST. Intermediary (I) susceptibility of pathovars was classified as resistant (R). A double synergy test was used for ESBL-producing strains testing. This consisted of placing discs (2-3 cm diameter) of ceftriaxone and cefotaxime around an amoxicillin-clavulanic acid disc on the bacterial plate.
8
Antibiotic Susceptibility Profiling
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Antibiotic susceptibility was determined on Mueller-Hinton agar (Liofilchem, Italy) using the standard disc diffusion procedure as described by the European Committee of Antimicrobial Susceptibility Testing (EUCAST) 18 . Nineteen antibiotics belonging to 7 different families (Table 2) were tested : amoxicillin (25 µg), amoxicillin-clavulanic acid (20/10 µg), ceftriaxone (30 µg), cefotaxime (30 µg), cefepime (30 µg), cefixime (10 µg), piperacillin (75 µg), piperacillin-tazobactam (100 +10 µg), imipenem (10 µg), tetracycline (30 µg), chloramphenicol (30 µg), trimethoprimsulfametoxazole (1.25 ± 23.75 µg), aztreonam (30 µg), colistin sulfate (50 µg), ciprofloxacin (5 µg), nalidixic acid (30 µg), gentamicin (15 µg), netilmicin (10 µg) and tobramycin (10 µg) (Bio-Rad, France). The diameters of the antibiotic susceptibility halos were recorded according to the recommendations of EUCAST. Intermediate (I) susceptibility of pathovars was classified as resistant (R).
9
Antimicrobial Susceptibility Testing of E. coli
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Susceptibility testing was performed according to CLSI by the disc diffusion method for the following antimicrobial agents: amoxicillin-clavulanic acid, ampicillin, ceftazidime, cefoxitin, gentamicin, cefotaxime, amikacin, nalidixic acid, tobramycin, ciprofloxacin, kanamycin, imipenem, tetracycline, trimethoprim-sulfamethoxazole, and chloramphenicol (Bio-Rad, France) . (Wayne, 2014) . The E. coli isolates were examined for ESBL production by double-disk synergy method by using a disk of amoxicillin/ clavulanic acid with two antimicrobial disks (ceftazidime and cefotaxime) (Kaur et al., 2013), the ESBL producing isolates were selected for more examinations. E. coli ATCC 25922 was used as a control strain.
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