Anabaena variabilis NIES 23 and Scenedesmus acutus NIES 94 were obtained from the National Institute for Environmental Studies (NIES), Japan. M. aeruginosa KW was isolated from freshwater in Korea. The strains were grown in BG-11 liquid medium (pH 7.5) under 120 µmol Photons/m 2 /s 1 provided by cool white fluorescent tubes at 27 ± 1°C. Micrococcus luteus (KCTC 1056), Bacillus subtilis (KCTC 1022), Staphylococcus aureus (KCTC 1621), Staphylococcus aureus (KCTC 1916), and Escherichia coli (KCTC 2443) were obtained from Korean Collection for Type Cultures (KCTC), and Erwinia carotovora was provided by Dr. Seung-Hwan Park (KRIBB, Korea). P. polymyxa E681, which was isolated from the roots of winter barley in the Republic of Korea, was cultured in Katznelson and Lochhead medium (KL medium) [9] .
Staphylococcus aureus
Manufactured by Korean Collection for Type Cultures
Sourced in United States
Staphylococcus aureus is a species of Gram-positive bacteria. It is a spherical-shaped bacterium commonly found in the human respiratory tract and on the skin.
Automatically generated - may contain errors
Lab products found in correlation
Escherichia coli, by Korean Collection for Type Cultures (6 mentions)
Bacillus subtilis, by Korean Collection for Type Cultures (3 mentions)
Salmonella typhimurium, by Korean Collection for Type Cultures (2 mentions)
Staphylococcus epidermidis, by Korean Collection for Type Cultures (2 mentions)
Pseudomonas aeruginosa, by Korean Collection for Type Cultures (2 mentions)
Pseudomonas aeruginosa pao1, by Korean Collection for Type Cultures (2 mentions)
Acinetobacter baumannii, by Korean Collection for Type Cultures (1 mentions)
Zinc acetate zn ch3coo 2 2h2o, by Merck Group (1 mentions)
Trifluoroacetic acid, by Thermo Fisher Scientific (1 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Fujifilm (1 mentions)
Ampicillin, by Merck Group (1 mentions)
Gallic acid, by Merck Group (1 mentions)
Folin ciocalteu reagent, by Merck Group (1 mentions)
6 hydroxy 2 5 7 8 tetramethylchroman 2 carboxylic acid trolox, by Merck Group (1 mentions)
2 4 6 tris 1 pyridyl 5 triazine, by Merck Group (1 mentions)
Spectramax microplate reader, by Molecular Devices (1 mentions)
Usnic acid, by Merck Group (1 mentions)
Low molecular weight chitosan, by Merck Group (1 mentions)
Tripolyphosphate tpp, by Merck Group (1 mentions)
Cas 9012 76 4, by Merck Group (1 mentions)
10 protocols using staphylococcus aureus
1
Isolation and Cultivation of Diverse Microbes
2
Antimicrobial Susceptibility of Common Pathogens
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We purchased Escherichia coli (KCTC1682), Salmonella typhimurium (KCTC 1926), Staphylococcus aureus (KCTC 1621), Bacillus subtilis (KCTC 1021), and Propionibacterium acnes (KCTC 3314, KCTC 3220, KCTC 5527, and KCTC 5933) from the Korean Collection for Type Cultures (KCTC), Korea Research Institute of Bioscience & Biotechnology (Daejeon, Korea). S. aureus (CCARM 0027 and CCARM 3708) was obtained from the Culture Collection of Antimicrobial Resistant Microbes (Seoul, Korea). The MIC of phloretin against each of these bacteria was determined by broth microdilution assay [43 (link)]. We defined the MIC as the lowest concentration of phloretin that completely inhibited bacterial growth; this was calculated as the average of three independent measurements in the range of 0.5 to 512 µM. P. acnes was grown for 48 to 72 h at 37 °C on sheep blood agar under anaerobic conditions in a mixture of H2 and CO2 (95:5, v/v). Microbroth dilution using Muller–Hinton (MH) broth was used to determine the efficacy of phloretin against P. acnes. We diluted a bacterial suspension in MH broth to achieve turbidity equivalent to a McFarland standard of 0.5 (~1 × 108 cells/mL), yielding 1 × 104 cells/mL in each well after inoculation. Phloretin MICs were determined after incubation for 48 h at 37 °C.
3
Antimicrobial Susceptibility Screening
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Four Gram-negative bacteria species (Escherichia coli KCTC 1682, Acinetobacter baumannii KCTC 2508, Pseudomonas aeruginosa KCTC 1637 and Salmonella typhimurium KCTC 1926) and three Gram-positive bacteria species (Staphylococcus aureus KCTC 1621, Bacillus subtilis KCTC 3028 and Staphylococcus epidermidis KCTC 1917) were procured from the Korean Collection for Type Cultures (KCTC). Multi-drug-resistant Gram-negative bacteria (Salmonella typhimurium CCARM 8003 and 8007, Escherichia coli CCARM 1229 and 1238, Pseudomonas aeruginosa CCARM 2002 and 2095 and Acinetobacter baumannii CCARM 12035 and 12036) were obtained from the Culture Collection of Antibiotic-Resistant Microbes (CCARM). To determine MICs, bacteria were cultured in Luria-Bertani (LB) medium for overnight at 37 °C. An aliquot of the culture was incubated in 1% peptone media at 37 °C until mid-log phase. 100 μl of serial 2-fold diluted peptides were treated in 96 well plates and added to 100 μl of 2 × 106 CFU/ml bacterial suspensions in 1% peptone media for 16 h at 37 °C. The lowest concentration of peptide that completely inhibited bacterial growth was determined to be the MIC.
4
Microbial Cultivation and Metal Treatments
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Escherichia coli (KCTC1682), Pseudomonas aeruginosa PAO1 (KCTC1637), Listeria monocytogenes (KCTC3569), and Staphylococcus aureus (KCTC 1916) were purchased from the Korean Collection for Type Cultures (KCTC, Daejeon, Korea). Klebsiella pneumoniae (ATCC4352) was purchased from American Type Culture Collection (ATCC). Streptococcus mutans (KCCM 40105) and Candida albicans (KCCM11282) were purchased from the Korean Culture Center of Microorganisms (KCCM; Seodaemun-gu, Seoul, South Korea). Tryptic soy broth (TSB; Difco Laboratory Inc., Detroit, MI, USA) was used to grow P. aeruginosa, S. aureus, E. coli, L. monocytogenes, and Vibrio vulnificus. For culturing C. albicans, the potato dextrose broth (PDB) containing glucose (5 %) was used, whereas for culturing the S. mutans, the brain heart infusion (BHI) broth (Difco Laboratory Inc., Detroit, MI, USA) was used. A deMan Rogosa Sharpe Medium (MRS; Difco Laboratory Inc., Detroit, MI, USA) was used to cultivate LAB. Gold (III) chloride trihydrate (CAS# 16961-25-4; purity 99%) and zinc acetate (Zn(CH3-COO)2·2H2O) (CAS # 5970-45-6) were obtained from Sigma-Aldrich Co. (St. Louis, MO, USA). The temperature for the growth of all microbes was 37 °C.
5
Antioxidant and Antibacterial Assays
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All solvents used in this study were of high-performance liquid chromatography (HPLC) grade. Trifluoroacetic acid (extra-pure grade) was supplied by Alfa Aesar (Ward Hill, MA, USA). Quercetin-3,4′-O-diglucoside and quercetin-4′-O-monoglucoside were supplied by Polyphenols Laboratories AS (Sandnes, Norway). Gallic acid, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), 2,4,6-Tris (1-pyridyl)-5-triazine (TPTZ), ferric chloride, Folin Ciocalteu reagent and ampicillin used were purchased fromSigma–Aldrich (St. Louis, MO, USA), and 2,2-diphenyl-1-picrylhydrazyl from Wako Pure Chemicals (Osaka, Japan) were used for antioxidant assays. Stock solutions of quercetin (1 mg mL−1) and quercetin glucosides (4 mg mL−1) were prepared in methanol. All the solutions were stored at −20 °C and calibration standards were obtained by appropriate dilution of the stock solutions. The standard bacterial strains of the Gram negative bacteria E. coli (KCTC-1924), Pseudomonas aeruginosa (KCTC-2004), and of the Gram positive bacteria B. cereus (KCTC-1012) and Staphylococcus aureus (KCTC-1916) were obtained from the Korean Collection for Type Cultures (KCTC).
6
Antimicrobial activity assessment of bacterial strains
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The Gram-negative bacterial strain Escherichia coli (KCTC 1682) and Gram-positive bacterial strain Staphylococcus aureus (KCTC 1621) were purchased from the Korean Collection for Type Cultures (Jeongeup, Korea). Acinetobacter baumannii (KCCM 40203) were purchased from Korea Culture Center of Microorganisms (Seoul, Korea). Additionally, five carbapenem-resistant Acinetobacter baumanii C1–C5 (CRAB C1–C5), which have the OXA-23 gene with carbapenem-resistance were collected from the patients with CRAB bacteremia, who presented symptoms and signs of infection at Korea University Anam Hospital (Seoul, Korea) (IRB registration no. 2020AN0157). The minimum inhibitory concentrations (MIC) of the AMP and antibiotics against the various bacterial strains were assessed using the serial dilution method on Muller–Hinton (MH) media, as described previously [30 (link)]. In brief, the peptides at 128 μg·mL−1 and antibiotics at 512 μg·mL−1 were serially diluted to 1/2 and incubated with a bacterial suspension of 2 × 105 CFU·mL−1 in MH media at 37 °C for 16 h. Absorbance at 600 nm was measured using a SpectraMAX microplate reader (Molecular Devices, San Jose, CA, USA).
7
Bacterial Strain Cultivation and Quantification
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The bacterial strains of Staphylococcus aureus (KCTC: 1621), Bacillus cereus (KCTC: 3624), methicillin-resistant Staphylococcus aureus (MRSA) (#77, #78, #79, and #80) and methicillin-susceptible Staphylococcus aureus (MSSA) (#85, #86, #87, and #88) were supplied by the Korean Collection for Type Cultures (KCTC), KRIBB (Korea) and BioNano Health Guard Research Center (H-GUARD) (Korea). All bacterial stains were cultured with LB broth (USA) at 37 °C. The bacterial concentration of S. aureus was determined by measuring the optical density (OD) at 600 nm (an OD600 of 1.0 Au = 8.8 × 108 CFU/mL).
8
Antimicrobial activity of usnic acid nanoparticles
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Escherichia coli (KCTC1682), Listeria monocytogenes (KCTC3569), Pseudomonas aeruginosa PAO1 (KCTC1637), and Staphylococcus aureus (KCTC 1916) were obtained from the Korean Collection for Type Cultures (KCTC, Daejeon, Korea). The growth media for the cultivation of L. monocytogenes, S. aureus and P. aeruginosa was tryptic soy broth (TSB; Difco Laboratory Inc., Detroit, MI, USA), whereas, for E. coli, Luria Bertani (LB) broth was used. Usnic acid, Tween 60, low molecular weight chitosan with 75–85% degree of deacetylation (CAS # 9012-76-4), and tripolyphosphate (TPP) were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA).
9
Antibacterial Activity of Casein Hydrolysates
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Antibacterial activity evaluated using a paper disc method [14] . Pathogenic bacteria Escherichia coli KCTC1682, Salmonella enterica KCTC2054, Bacillus cereus KCTC362A and Staphylococcus aureus KCTC3881 obtained from the Korean Collection for Type Cultures (KCTC) and it dispensed on Mueller hinton agar (Difco™, USA). One hundred microliter of the pathogenic bacteria culture (10 3 -10 4 CFU/mL) uniformly distributed by pouring 30 mL of pre-sterilized Mueller hinton agar and allowed for solidification.
A paper disc method (Advantec, Japan), impregnated with the compound, placed on the surface of the Mueller hinton agar. The 100 μL of casein hydrolysate fractions dispensed into each paper disc and incubated in the incubator for 37℃. After incubation for 24 hours, the diameter (mm) of the inhibition clear zone around the well measured and compared to each other.
A paper disc method (Advantec, Japan), impregnated with the compound, placed on the surface of the Mueller hinton agar. The 100 μL of casein hydrolysate fractions dispensed into each paper disc and incubated in the incubator for 37℃. After incubation for 24 hours, the diameter (mm) of the inhibition clear zone around the well measured and compared to each other.
10
Skin Microbiome Diversity Analysis
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The six strains used in this study are common skin microflora. They included four gram-positive strains and two yeasts. Malassezia furfur (KCTC 7743) and Malassezia globosa (KCTC 7846) , which cause dandruff, Corynebacterium xerosis (KCTC 9105) , which causes underarm malodor, and Staphylococcus aureus (KCTC 6538) and Staphylococcus epidermidis (KCTC 13170) , which are typical skin microorganisms, were obtained from the Korean Collection for Type Cultures (KCTC, Jeongeup, Jeonbuk) . Corynebacterium jeikeium (ATCC 43734) was obtained from the American Type Culture Collection (ATCC, Manassas, VA) . Before use, the Malassezia species were each inoculated on Leeming and Notman (LN) medium and incubated for 1 d in orbital shakers at 30℃. The other four strains of bacteria were inoculated onto Brain Heart Infusion (BHI, BD, Sparks, MD, USA) media and incubated for 1 d in an orbital shaker at 35℃.
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