MICs of ampicillin, mecillinam, nitrofurantoin, sulphonamide, trimethoprim, tetracycline, nalidixic acid, ciprofloxacin, kanamycin and chloramphenicol were measured by agar dilution on iso-sensitest medium (Oxoid, Basingstoke, UK). MIC was defined as the lowest concentration that completely inhibited growth.
Iso sensitest medium
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
Iso-Sensitest medium is a microbiological culture medium used for susceptibility testing of bacteria. It is formulated to support the growth of a wide range of microorganisms.
Automatically generated - may contain errors
Lab products found in correlation
Mic test strip, by Liofilchem (1 mentions)
Ampicillin, by bioMérieux (1 mentions)
Erythromycin, by bioMérieux (1 mentions)
Chloramphenicol, by bioMérieux (1 mentions)
Clindamycin, by bioMérieux (1 mentions)
Gentamicin, by bioMérieux (1 mentions)
Tetracycline, by bioMérieux (1 mentions)
Vancomycin, by bioMérieux (1 mentions)
Streptomycin, by bioMérieux (1 mentions)
Kanamycin, by bioMérieux (1 mentions)
4 protocols using iso sensitest medium
1
Antimicrobial Susceptibility Testing
2
Lactobacillus plantarum P-8 cultivation
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Lactobacillus plantarum P-8 was obtained from the Key Laboratory of Dairy Biotechnology and Engineering, Ministry of Education, Inner Mongolia Agricultural University. It was cultivated either in de Mann-Rogosa-Sharpe (MRS) broth (catalog no. CM0359, Oxoid) or in LAB susceptibility test medium (LSM) (41 (link), 42 (link)). The LSM consisted of 90% Iso-Sensitest medium (catalog no. CM0473; Oxoid) and 10% MRS broth. The E. coli DH5α strain was used for standard cloning, and the culture was grown aerobically in Luria-Bertani broth at 37°C. The antibiotics chloramphenicol (10 μg/ml for both E. coli and L. plantarum P-8) and erythromycin (250 μg/ml and 5 μg/ml for E. coli and L. plantarum P-8, respectively) were used for selecting genetic mutants.
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3
Antibiotic Resistance Profiling of Lactic Acid Bacteria
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An antibiotic resistance of the strains MG5206 and MG5232 was assayed using the minimum inhibitory concentration (MIC) test strip method. The pellets of each strain were harvested by centrifugation (3460× g, 10 min) after culturing in MRS broth at 37 °C for 18 h, washed twice with phosphate-buffered saline (PBS, pH 7), and resuspended in PBS to obtain a resuspended solution with a McFarland turbidity level of 0.5. The suspended solution was inoculated in a lactic acid bacteria susceptibility test medium (LSM) plate, with a mixture of 90% Iso-Sensitest medium (Oxoid Ltd., Hampshire, UK) and 10% MRS with 1.5% agar, using swabs [19 (link)]. The plates were dried for 10 min, and MIC test strips (Liofilchem Inc., Roseto degli Abruzzi, Italy) were placed on the plate according to the manufacturer’s instructions and incubated at 37 °C, and the results were recorded 24 h after inoculation. Antibiotic susceptibility was determined according to the European Food Safety Authority (EFSA) guidelines [20 (link)].
4
Antibiotic Susceptibility Testing of Lactic Acid Bacteria
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E‐test gradient strips of gentamicin, kanamycin, streptomycin, tetracycline, erythromycin, clindamycin, chloramphenicol, ampicillin or vancomycin were used (BioMerieux, France). The choice of nine antibiotics was performed according to the EFSA guidelines on Additives and Products or Substances used in Animal Feed (EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) 2012 ). The E‐test was performed following the manufacturer's instructions using LMS‐agar medium composed of MRS and Iso‐Sensitest medium (Oxoid, UK) with weight proportions of 10% and 90%, respectively. The inoculum was prepared by selecting three o/n colonies grown on MRS‐agar and suspending them separately in 0.5 ml of sterile 0.9% saline. Drops of the resulting solutions were added to 5 ml of sterile 0.9% saline to adjust the turbidity to 0.5 according to the McFarland standard. Bacteria were spread on LMS‐agar surface with a swab soaked with the final cell suspension. Subsequently, the strips were laid on the LMS‐agar‐bacteria surface and incubated at 37°C for 48 hr. The MIC was determined from the inhibition curves that intersected the scale on a strip.
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