The brucells isolates were tested for their susceptibility to 11 antibiotics (Rifampicin; Ciprofloxacin; Ampicillin; Erythromycin; Novobiocin; Kanamycin; Gentamicin; Streptomycin; Tetracycline; Doxycycline and Carbenicillin), obtained from Himedia Laboratories. Testing was performed on Mueller-Hinton Agar plates using the Kirby-Bauer disk diffusion technique (Bauer & Kirby, 1966 (link)). The antibiotic resistance of each brucella isolate was determined based on the breakpoints of the inhibition zone diameters for individual antibiotic agents and as recommended by the disk manufacturer.
Carbenicillin
Manufactured by HiMedia
Sourced in India, United Kingdom
Carbenicillin is a broad-spectrum antibiotic used in research laboratories. It belongs to the group of penicillin-like antibiotics and is effective against a variety of Gram-positive and Gram-negative bacteria. Carbenicillin is commonly used in cell culture and molecular biology applications to prevent bacterial contamination.
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Lab products found in correlation
Ciprofloxacin, by HiMedia (9 mentions)
Gentamicin, by HiMedia (7 mentions)
Amikacin, by HiMedia (6 mentions)
Piperacillin tazobactam, by HiMedia (6 mentions)
Cotrimoxazole, by HiMedia (4 mentions)
Polymixin b, by HiMedia (4 mentions)
Erythromycin, by HiMedia (3 mentions)
Streptomycin, by HiMedia (3 mentions)
Tetracycline, by HiMedia (3 mentions)
Ceftazidime, by HiMedia (3 mentions)
Ampicillin, by HiMedia (2 mentions)
Netilmicin, by HiMedia (2 mentions)
Faropenem, by HiMedia (2 mentions)
Cefamandole, by HiMedia (2 mentions)
Doxycycline hydrochloride, by HiMedia (2 mentions)
Pefloxacin, by HiMedia (2 mentions)
Chloramphenicol, by HiMedia (2 mentions)
Nalidixic acid, by HiMedia (2 mentions)
Amoxycillin clavulanic acid, by HiMedia (2 mentions)
Mueller hinton agar (mha), by HiMedia (2 mentions)
10 protocols using carbenicillin
1
Antibiotic Susceptibility of Brucella Isolates
2
Antibiotic Susceptibility of NDM-1 Harboring Strains
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Antibiotic susceptibility of blaNDM-1 harboring parent strains, transformants and transconjugants were determined by Kirby Bauer disc-diffusion method including piperacillin-tazobactam (100/10 μg), co-trimoxazole (25 μg), amikacin (30 μg), gentamicin (10 μg), ciprofloxacin (5 μg), polymyxin B (300units), netilmicin (30 μg), carbenicillin (100 μg), tigecycline(30 μg) and faropenem (5 μg) (Hi-Media, Mumbai, India). MICs of imipenem, meropenem, ertapenem, cefepime, aztreonam, gentamicin, amikacin, ciprofloxacin, piperacillin-tazobactam & polymixin-B were determined for parent strains harboring blaNDM-1, as well as transformants and transconjugants by agar dilution method. Each stock solution for the corresponding antibiotic was made at 1 mg/ml concentration in nuclease free water and was stored at −80 °C. The quality control for stock solution was checked each time against E. coli ATCC 25922. The result of the susceptibility testing was interpreted as per CLSI guidelines [24 ]. However, for polymyxin B, faropenem and carbenicillin, the organisms were considered as non susceptible if the MIC value was higher and diameter of the zone of inhibition was lower than the values given in CLSI guidelines for respective antibiotics against E. coli ATCC 25922.
3
Evaluating Natural Compound Antimicrobial Synergies
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Natural compounds – diadzein, lanatoside C, protocatechuic acid, scopolamine, gentisic acid, umbelliferone and MC-207,110 were purchased from Sigma-Aldrich (India). Cyanide-m-chlorophenylhydrazone (CCCP), ethidium bromide, carbenicillin and levofloxacin were purchased from Hi-Media, India. Greiner Bio-one flat bottomed black 96-well plate was purchased from Sigma-Aldrich. P. aeruginosa MexAB-OprM harbouring strain was kindly provided as a gift by Dr. Keith Poole, Queen's University, Canada and E. coli AcrAB-TolC harbouring strain was kindly provided as a gift by Dr. Klaas Martinus Pos, Goethe University, Germany. MIC and checkerboard assays were performed in the Cation adjusted Mueller Hinton Broth (CA-MHB) purchased from Hi-Media, India. Luria-Bertani (LB) broth purchased from Hi-media, India was used for ethidium bromide accumulation studies.
4
Antibiotics Susceptibility of KPC-2
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The antibiotic susceptibility was done by Kirby Bauer disc-diffusion method against antibiotics viz. piperacillin-tazobactam (100/10 μg), amikacin (30 μg), gentamicin (10 μg), ciprofloxacin (5 μg), polymixin B (300 units), netilmicin (30 μg) and carbenicillin (100 μg) (Hi-Media, Mumbai, India). Minimum inhibitory concentration was performed by agar dilution method against imipenem, ertapenem, meropenem, cefepime & aztreonam and the results were compared with standard CLSI guidelines [14 ]. The antibiotic susceptibility of the transformants and clone of blaKPC-2 was also determined.
5
Antimicrobial Susceptibility Testing Protocol
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Antimicrobial susceptibility testing was performed according to the protocols of the CLSI [44 ]. The results were evaluated according to the EUCAST cut-off values [45 ] using antibiotics applicable to the treatment of patients, namely meropenem (10 µg, MEM10C) from Mast Group Ltd., UK, penicillin-G (10 U, SD028-1PK), ampicillin (10 µg, SD002-1PK), amoxycillin (25 µg, SD129-1PK), amoxycillin/clavulanic acid (20/10 µg, AUG30C), carbenicillin (100 µg, SD004-1PK), cefamandole (30 µg, SD200-1PK), erythromycin (15 µg, SD013-1PK), clarithromycin (15 µg, SD192-1PK), streptomycin (10 µg, SD031-1PK), tetracycline (30 µg, SD037-1PK), doxycycline hydrochloride (30 µg, SD012-1PK), chloramphenicol (30 µg, SD006-1PK), nalidixic acid (30 µg, SD021-1PK), ciprofloxacin (5 µg, SD060-1PK), pefloxacin (5 µg, SD070-1PK), and co-trimoxazole (25 µg, SD010-1PK) from HiMedia, India.
6
Antimicrobial Susceptibility Testing of E. coli
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Antimicrobial susceptibility testing was performed via a standard disk diffusion method, also known as the Kirby–Bauer method, according to the protocols of the CLSI [36 ]. We again used antibiotics applicable to the treatment of patients, namely meropenem (10 µg, MEM10C Oxoid ltd, Basingstoke, Hampshire, UK), ampicillin (10 µg, SD002-1PK), amoxycillin (25 µg, SD129-1PK), amoxycillin/clavulanic acid (20/10 µg, AUG30C), carbenicillin (100 µg, SD004-1PK), cefamandole (30 µg, SD200-1PK), erythromycin (15 µg, SD013-1PK), streptomycin (10 µg, SD031-1PK), tetracycline (30 µg, SD037-1PK), doxycycline hydrochloride (30 µg, SD012-1PK), chloramphenicol (30 µg, SD006-1PK), nalidixic acid (30 µg, SD021-1PK), ciprofloxacin (5 µg, SD060-1PK), pefloxacin (5 µg, SD070-1PK) and co-trimoxazole (25 µg, SD010-1PK) from HiMedia, India. The results were evaluated according to the cut-off breakpoint values of EUCAST version 12.0, 2022 [37 ], CLSI, 31st edition [36 ] and the Manual of BBL Products and Laboratory Procedures [38 ]. Breakpoint values of erythromycin for other bacterial species were taken for E. coli.
7
Antibiotic Susceptibility Testing Protocol
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Antibiotic susceptibility testing was performed on Mueller-Hinton agar (Himedia, Mumbai, India) plates by Kirby-Bauer disc diffusion method and interpreted as per CLSI recommendations [15 ]. The antibiotics tested viz., ciprofloxacin (5μg), amikacin (30μg), gentamicin (10μg), carbenicillin (10μg), polymixin B (300 μg), ceftazidime (30 μg), Piperacillin-Tazobactam (100/10 μg) (Himedia, Mumbai, India).
Minimum inhibitory concentration (MIC) was determined on Muller Hinton Agar plates by agar dilution method against imipenem and meropenem (Himedia, Mumbai, India) according to CLSI guidelines and interpreted according to CLSI breakpoint [15 ].
Minimum inhibitory concentration (MIC) was determined on Muller Hinton Agar plates by agar dilution method against imipenem and meropenem (Himedia, Mumbai, India) according to CLSI guidelines and interpreted according to CLSI breakpoint [15 ].
8
Antibiotic Susceptibility Testing by Disc Diffusion
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Antibiotic susceptibility testing was performed on Mueller Hinton agar (Himedia, Mumbai, India) plates by Kirby-Bauer disc diffusion method and interpreted as per CLSI recommendations [10 ]. The antibiotics tested viz., meropenem (10μg) ciprofloxacin (5μg), amikacin (30μg), gentamicin (10μg), carbenicillin (10μg), polymixin B (300 μg), ceftazidime (30 μg), Piperacillin-Tazobactam (100/10 μg), faropenem (5μg) (Himedia, Mumbai, India).
Minimum inhibitory concentration (MIC) was determined on Muller Hinton Agar plates by agar dilution method against meropenem and ciprofloxacin (Himedia, Mumbai, India) according to CLSI guidelines and interpreted according to CLSI breakpoint [10 ].
Minimum inhibitory concentration (MIC) was determined on Muller Hinton Agar plates by agar dilution method against meropenem and ciprofloxacin (Himedia, Mumbai, India) according to CLSI guidelines and interpreted according to CLSI breakpoint [10 ].
9
Antimicrobial Susceptibility of Bacterial Isolates
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Disc diffusion method was performed for antimicrobial susceptibility testing of the test isolates and the results were interpreted using CLSI breakpoints. The antibiotics tested includes- ciprofloxacin (5 μg), amikacin (30 μg), cefepime (30 μg), aztreonam (30 μg), ceftriaxone (30 μg), cotrimoxazole (25 μg), ceftazidime (30 μg), levofloxacin (5 μg) gentamicin (10 μg), carbenicillin (10 μg), ceftazidime (30 μg) and Piperacillin/Tazobactam (100/ 10 μg) (Hi-media, Mumbai, India). The minimum inhibitory concentrations (MICs) of carbapenems (meropenem, ertapenem and imipenem), colistin and polymixin B were determined by agar dilution method and the results were interpreted as per CLSI [25 ] and EUCAST guidelines (Version 9.0) respectively. Escherichia coli ATCC 25922 was used as control.
10
Antibiotic Susceptibility Profiling
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The antibiotic susceptibility was done by Kirby Bauer disc diffusion method against antibiotics as piperacillin-tazobactam (100/10 μg), amikacin (30 μg), gentamicin (10 μg), ciprofloxacin (5 μg), polymixin B (300 units) ampicillin (30 μg), cotrimoxazole (10 μg) and carbenicillin (100 μg) (Hi-Media, Mumbai, India). Minimum inhibitory concentration was performed by agar dilution method against imipenem, meropenem, cefepime, aztreonam, cotrimoxazole, ampicillin, & ciprofloxacin and the results were compared with standard CLSI guidelines [10] . The antibiotic susceptibility of the transformants was also determined. Flow diagram of the whole procedure carried out is given in Figure 1.
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