Antibiotic susceptibility testing was carried out following the EUCAST guidelines [29 (link)] as closely as possible (variations included the use of 90 mm plates, stacking during incubation, and inoculation with only one antibiotic disc/plate to prevent overlap if bacterial susceptibility increased). The antibiotics used were chloramphenicol, ciprofloxacin, erythromycin, fusidic acid, gentamicin, mupirocin, oxacillin, rifampicin, tetracycline and vancomycin. These 10 were selected as they represented antibiotics, with differing antimicrobial mechanisms, which are recommended clinical treatment options for Staphylococcus aureus infections. Fresh bacterial cultures were diluted to a 108 CFU/ml population, spread onto mueller-hinton (MH) agar plates (Oxoid, UK) using sterile swabs and a single antibiotic disc (Mast Group, UK) placed into the centre of the plate. The plates were immediately incubated at 35 ± 2 °C for 18–20 h after which the diameter of the zone of inhibition was measured in normal laboratory lighting and recorded to the nearest mm.
Mh agar plates
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
MH agar plates are a type of microbiological growth medium used for the cultivation and isolation of a wide range of bacterial species. They provide a suitable environment for the growth of microorganisms, facilitating their identification and further analysis.
Automatically generated - may contain errors
Lab products found in correlation
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (1 mentions)
Thiazolyl blue tetrazolium bromide, by Merck Group (1 mentions)
α terpineol, by Merck Group (1 mentions)
Butylated hydroxytoluene, by Merck Group (1 mentions)
α pinene, by Merck Group (1 mentions)
1 8 cineole, by Merck Group (1 mentions)
Muller hinton agar, by Bio-Rad (1 mentions)
Red violet bile glucose medium, by Thermo Fisher Scientific (1 mentions)
Cefotaxime, by Merck Group (1 mentions)
Mueller hinton media, by Bio-Rad (1 mentions)
5 protocols using mh agar plates
1
Antibiotic Susceptibility Testing of Staphylococcus aureus
2
Assessing EDTA's Effect on Imipenem
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Experimental strains were diluted to 0.5 M bacterial suspension using sterilized saline water and then evenly coated onto Müller-Hinton (M-H) agar plates (OXOID, UK). Two 10 µg IPM drug sensitivity test papers (OXOID, UK) were pasted. One of the test papers had 10 µL EDTA at a concentration of 0.5 mmol/l (pH 8.0). The M-H plates with pasted drug sensitivity test papers were incubated at 35°C for 18 h. Next, the diameter of the inhibition zone of IPM drug sensitivity test paper with EDTA was compared to that with IPM.
3
Antimicrobial and Antioxidant Evaluation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
α‐Pinene, α‐terpineol, 1,8‐cineole, butylated hydroxytoluene (BHT), thiazolyl blue tetrazolium bromide (MTT), and 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) were purchased from Sigma‐Aldrich Co. Muller–Hinton agar was purchased from Bio‐Rad. MH agar plates and red–violet bile glucose medium were obtained from Oxoid Ltd.
4
Conjugation Assay: Tracking Antibiotic Resistance
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In-vitro conjugation assays were performed in triplicate, using filter-mating, to investigate the potential of the four OXA-244-producing E. coli ST38 strains for transferring ARGs to a green fluorescent protein (GFP)-tagged E. coli strain CV601, according to a previously described method [22 22. Jutkina, J. ]. Equal volumes of the prepared donors and recipient were mixed, and the conjugation mixtures were vacuum-filtered on to S-Pak filters (pore size 0.22 μm) (Millipore, USA), before the filters were placed on MH agar plates (Oxoid, UK). The plates were incubated at 30 °C for 3 h. The filters were transferred to tubes containing 10 mL sterile saline (0.85% NaCl) and glass beads and vortexed for 1 min. The samples were serially diluted with sterile saline (0.85% NaCl) before plating 100-μL aliquots on MH Orientation (CHROMagar™, France) chromogenic media with 50 μg/mL kanamycin, 50 μg/mL rifampicin and 2 μg/mL cefotaxime (Sigma-Aldrich, Germany), for selection of transconjugants. In addition, MH plates with 50 μg/mL kanamycin and 50 μg/mL rifampicin were used to estimate the number of recipients. The plates were incubated at 37 °C overnight. Transconjugants were identified based on GFP expression when exposed to UV light.
5
Antibacterial Assay of LmPS Extract
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibacterial tests were performed as described by Ben Hsouna et al. [5 (link)] and the broth microdilution test by the sterile Mueller–Hinton media (Bio-Rad, Marnes-la-Coquette, France). One hundred μL was evenly spread on the surface of MH agar plates (Oxoid Ltd., Basingstoke, UK). Wells were dug into the agar using a sterile Pasteur pipette. The final concentration was 50 mg/mL of LmPS dissolved in distilled water. Then, 50 μL of extract was placed into the wells and the plates were incubated for 24 h at 37 °C. Streptomycin (20 μg/well) and distilled water were used as positive and negative controls, respectively. The antimicrobial effect was checked measuring the diameter of the circular inhibition zones of the wells. The tests were carried out in triplicate.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!