Muller-Hinton was purchased from Merck, Germany. Phosphate-buffered saline (PBS) and Hanks’ balanced salt solutions (HBSS) were purchased from Solarbio Science and Technology, China. Glutaraldehyde and [3-(4,5-dimethyl thiazol-2yl)-2,5-diphenyl tetrazolium bromide] (MTT) were purchased from Sinopharm (Beijing, China). The University of Malaya College of Medicine, Department of Pharmacy Center for Natural Product Research and Drug Discovery Department of Pharmacology Faculty of Medicine University of Malaya Kuala Lumpur Malaysia, provided the normal liver cell line WRL 68 Cell and the cancer cell line MCF-7. Cancer cells were nurtured and cultivated, and tests were performed on them at Al-Nahrain University’s Biotechnology Research Center.
Muller hinton
Manufactured by Merck Group
Sourced in Germany
Muller-Hinton is a type of growth media used in microbiology laboratories. It is commonly used for antimicrobial susceptibility testing, such as the Kirby-Bauer disk diffusion method, to determine the sensitivity of bacteria to various antibiotics.
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Lab products found in correlation
Hanks balanced salt solutions, by Solarbio (1 mentions)
Glutaraldehyde, by Sinopharm (1 mentions)
Phosphate buffered saline (pbs), by Solarbio (1 mentions)
Nutrient broth, by Merck Group (1 mentions)
κ carrageenan, by Merck Group (1 mentions)
Purified soy lecithin, by Lipoid (1 mentions)
Phosphate buffer saline (pbs), by Merck Group (1 mentions)
Glycerol, by Daejung Chemicals (1 mentions)
Gelred, by Biotium (1 mentions)
Phthalic anhydride, by Merck Group (1 mentions)
Sodium ascorbate, by Merck Group (1 mentions)
Hydrazine monohydrate, by Merck Group (1 mentions)
1 1 carbonyldiimidazole, by Merck Group (1 mentions)
Copper 2 acetate, by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Merck Group (1 mentions)
Acetic acid, by Merck Group (1 mentions)
Ethylenediaminetetraacetic acid, by Merck Group (1 mentions)
Dimethylformamide, by Merck Group (1 mentions)
Tetrahydrofuran, by Merck Group (1 mentions)
Brain heart broth, by Merck Group (1 mentions)
7 protocols using muller hinton
1
Cytotoxicity Evaluation of Natural Products
2
Antibiotic Susceptibility of E. coli and Klebsiella
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Sensitivity of E. coli and Klebsiella isolates to antibiotics of Gentamicin, Ciprofloxacin, Cotrimoxazole, Clavulanate/amoxicillin, Imipenem, Piperacillin, Cefepime, Ceftazidime, and Cefotaxime was measured by the disk diffusion method (Kirby-Bauer) according to CLSI protocols, using bacterial suspension with a dilution equivalent to opacity of half pipe McFarland and the solid medium of Muller-Hinton (Merck Company, Germany). For this purpose, a bacterial suspension with a dilution equivalent to opacity of half pipe McFarland was prepared (1.5×108 bacteria). Then, the suspension was inoculated on the plates containing Müller-Hinton agar medium and cultured with GRASS method. Antibiotics disks were put on the medium using sterile forceps, as they had a distance of 20 mm from each other and 15 mm from the wall of plate. The plates were incubated for 18 hours at a temperature of 37°C. After the incubation period, the diameter of areola related to lack of bacterial growth around the disc, was measured using a ruler (in mm) and compared to the table provided in CLSI protocols and then sensitivity (s), intermediate (I), and resistant (R) were qualitatively reported. All tests were performed on the standard strain of E. coli ATCC25922 as the control.
3
Formulation of Antimicrobial Rhamnolipid Hydrogel
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Rhamnolipids (R90) were purchased from AGAE Technologies (USA). Phosphate buffer saline and) κ-carrageenan was purchased from Sigma-Aldrich (Germany). Hydroxypropyl methylcellulose (HPMC) and glycerol were purchased from Daejung chemicals (South Korea). Nutrient broth, Muller Hinton, and brain heart infusion (BHI) broth/agar were purchased from Merck (USA). Nisin Z was purchased from Honghao Chemical Co. (Shanghai, China). Purified soy lecithin (phosphatidylcholine ⩾ 94%) was provided by Lipoid (Ludwigshafen, Germany).
4
Antimicrobial Susceptibility Testing by Disk Diffusion
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Antimicrobial susceptibility of the isolates was determined by the Kirby-Bauer disk diffusion method on Muller-Hinton (Merck, Germany) agar according to Clinical and Laboratory Standards Institute (CLSI) guideline (17 (link)). Briefly, a suspension of each isolates was adjusted to a turbidity equivalent to 0.5 McFarland and inoculated on Muller-Hinton agar plate. The tested antimicrobial agents were as follows: piperacillin (100 μg), ceftazidime (30 μg), meropenem (10 μg), imipenem (10 μg), gentamicin (10 μg), amikacin (30 μg), ofloxacin (30 μg), cefotaxime (30 μg), colistin (10 μg), ciprofloxacin (5 μg), tetracycline (75 μg) and aztreonam (30 μg) (MAST, Group Ltd, Merseyside, UK) (8 (link), 13 (link)). E. coli ATCC 25922 and P. aeruginosa ATCC 27853 were used as standards in disk diffusion. The plates were incubated overnight at 35°C and the results were interpreted as susceptible, intermediate or resistant according to the criteria recommended by the CLSI (2 (link)). MDR isolates were defined if they showed simultaneous resistance to one agent from three antibiotics group (18 (link)).
5
Bacterial Plasmid DNA Extraction
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Cecal sample (0.01 g) was mixed with 0.1 mL of 0.85% NaCl. The 0.1 mL mix was spread on a non-selective Muller-Hinton (Merck) agar plate and incubated overnight at 37°C. All bacterial growth on the plate was scraped off, inoculated into 6 mL of Muller-Hinton broth and incubated overnight at 37°C with shaking at 225 rpm. Plasmid DNA was extracted from this bacterial culture using the Macherey-Nagel NucleoSpin Plasmid Kit according to the manufacturer's guidelines. The resulting DNA samples were visualized on a 1% agarose gel stained with GelRed (Biotium) and run at 70 volts (V) for 60 minutes (min).
6
Chitosan-based Antimicrobial Agent Synthesis
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Low molecular weight (LMW) chitosan powder, 85% deacetylated was purchased from Sigma-Aldrich (USA). Penta-sodium tripolyphosphate (TPP), Muller-Hinton (MH) broth, and agar were supplied by Merck (Germany). Acetic acid, sodium hydroxide, phthalic anhydride, dimethyl formamide (DMF), tetrahydrofuran (THF), 1,1′-carbonyldiimidazole (CDI), proparglyamine, copper (II) acetate, sodium ascorbate, 1-azidoadamantane, azidobenzene solution, 1-azidomethyl-2-methyl benzene solution, azidomethyl phenyl sulphide, 2-azidomethyl-1-Boc-pyrrolidine, ethylenediaminetetraAcetic acid (EDTA), hydrazine monohydrate were also supplied by Sigma-Aldrich (USA). All chemicals and reagents were of highest analytical grade and used directly without further purification.
7
Silver Nanoparticle Synthesis and Antimicrobial Testing
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Silver nitrate, ACS reagent, 99+% (AgNO3) for AgNP synthesis were obtained from Sigma-Aldrich, for appropriate microorganisms culturing Muller Hinton (MHB), Sabaroud Dextrose Broth (SDB) and Brain Heart Broth (BHB) were obtained from Merck. All microorganism strains were kept at −20˚C in the appropriate medium containing 10% glycerol and regenerated twice prior to the manipulations. For anticancer analyses all chemicals were purchased from Sigma Aldrich, Germany. Freshly prepared doubly distilled water was used throughout the experimental work.
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