Phenolic and carotenoid components’ standards were supplied by Sigma-Aldrich (Darmstadt, Germany). Plant Flavonoids Colorimetric Assay Kit was obtained from Elabscience Biotechnology Inc. (Houston, TX, USA). Prior to HPLC screening, each of the analyzed samples underwent filtration using a 0.45 µm MF-Millipore™ Membrane Filter acquired from Merck (Darmstadt, Germany). The bacterial reference strains: Staphylococcus aureus ATCC 25923, Staphylococcus aureus ATCC 700699, Bacillus cereus ATCC 14579, Enterococcus faecalis ATCC 29219, Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853 were purchased from Oxoid Ltd. (Hampshire, UK). Culture mediums for bacteria were purchased from Merck (Darmstadt, Germany). Sigma-Aldrich (St. Louis, MO, USA) provided the enzymatic mixtures for cell cultures, while Gibco Life Technologies (Paisley, UK) provided the culture medium constituents. Thermo Fisher Scientific Inc. (Waltham, MA, USA) provided the Annexin V-FITC with propidium iodide (PI) flow-cytometry Kit.
Staphylococcus aureus atcc 25923
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
Staphylococcus aureus ATCC 25923 is a strain of the bacterium Staphylococcus aureus. It is a gram-positive, non-spore-forming, coagulase-positive coccus. This strain is commonly used as a quality control and reference strain in microbiological testing and research.
Automatically generated - may contain errors
Lab products found in correlation
Escherichia coli atcc 25922, by Thermo Fisher Scientific (5 mentions)
Pseudomonas aeruginosa atcc 27853, by Thermo Fisher Scientific (4 mentions)
Mf millipore membrane filter, by Merck Group (1 mentions)
Plant flavonoids colorimetric assay kit, by Elabscience (1 mentions)
Annexin 5 fitc, by Thermo Fisher Scientific (1 mentions)
Catalase solution, by Merck Group (1 mentions)
Phenol, by Merck Group (1 mentions)
Mueller hinton, by Thermo Fisher Scientific (1 mentions)
Sheep blood, by Thermo Fisher Scientific (1 mentions)
Todd hewitt broth, by Thermo Fisher Scientific (1 mentions)
Columbia agar plate, by Thermo Fisher Scientific (1 mentions)
7 protocols using staphylococcus aureus atcc 25923
1
Phytochemical and Antimicrobial Evaluation
2
Antibacterial Activity of Honey
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Antibacterial activity of honey was determined as described in Irish, Blair, and Carter (2011) (link). Using Staphylococcus aureus ATCC 25923 (Oxoid, Hampshire, UK) as the test organism, honey solutions were prepared fresh to a 50% (w/v) solution in sterile water and further diluted to 25% (v/v) in either sterile water to test for total activity, or a 2 mg/ml catalase solution (3187 units/mg; Sigma-Aldrich, St. Louis, USA) for non-peroxide activity. Phenol (Sigma-Aldrich, St. Louis, USA) standards of 2%−7% v/v were prepared fresh in sterile water.
Aliquots (100ul) of each honey sample and Phenol solutions were dispensed into the wells of assay plates in duplicate using quasi-Latin squares to randomise sample distribution. Plates were incubated (37 °C, 18–24 h) before the diameter of each zone of inhibition was measured using digital Vernier callipers (Kinechrome, Carole Park, Australia). The mean diameter of the zone of inhibition was calculated and squared, and a standard curve (concentration of Phenol versus mean squared diameter) was generated using the Phenol standards. The activity (total and non-peroxide) of each honey was calculated using the standard curve, multiplied by 4.69 to account for the dilution and density of honey, and expressed as the equivalent Phenol concentration.
Aliquots (100ul) of each honey sample and Phenol solutions were dispensed into the wells of assay plates in duplicate using quasi-Latin squares to randomise sample distribution. Plates were incubated (37 °C, 18–24 h) before the diameter of each zone of inhibition was measured using digital Vernier callipers (Kinechrome, Carole Park, Australia). The mean diameter of the zone of inhibition was calculated and squared, and a standard curve (concentration of Phenol versus mean squared diameter) was generated using the Phenol standards. The activity (total and non-peroxide) of each honey was calculated using the standard curve, multiplied by 4.69 to account for the dilution and density of honey, and expressed as the equivalent Phenol concentration.
3
Antibiotic Susceptibility Profiling
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Susceptibility to antibiotics was carried out using the disk-diffusion method on Muller Hinton Agar at 37 °C for 18 h. Results were interpreted according to CLSI breakpoint criteria (2014). Three reference isolates (Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, and Pseudomonas aeruginosa ATCC 27853) were used for quality control of the purchased disks (Oxoid, Basingstoke, Hampshire, UK). S. aureus isolates were tested on a total of 10 different antibiotic disks: oxacillin (OX5), cefoxitin (FOX30), penicillin G (P10), spiramycin (SP100), clindamycin (DA2), lincomycin (L15), erythromycin (E15 (RD5), vancomycin (VA30), amikacin (AK30), and ofloxacin (OFX5), whereas Enterobacteriaceae members were tested for amoxicillin (AMX25), amoxicillin/clavulanate (AMC10), cefotaxime (CTX30), ceftriaxone (CRO 30), ampicillin (AMP10), cefazolin (CF30), imipenem (IMP10), ofloxacin (OFX5), amikacin (AK10), and trimethoprim/sulfamethoxazole (SXT1.25/23.75). Non-fermentative bacilli (P. aeruginosa and A. baumannii) were tested for the following antibiotics: piperacillin (PIP100), ticarcillin (TIC75), ticarcillin–clavulanate (TCC75), aztreonam (ATM30), ceftazidime (CAZ30), piperacillin (PRL100), levofloxacin (LEV5), amikacin (AK30), colistin (CT10), and trimethoprim/sulfamethoxazole (SXT1.25/23.75).
4
Antibiotic Susceptibility Testing of S. aureus
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The S. aureus isolates were subjected to the Kirby-Bauer disc diffusion susceptibility tests following the clinical and laboratory standard institute (CLSI) guidelines [55 ]. Briefly, 24 antibiotics (Oxoid, Thermo Fischer Scientific, Canada) relevant to human and veterinary health from the classes of ß-lactams, aminoglycosides, cephalosporins, quinolones, macrolides, lincosamide, tetracycline, chloramphenicol, and sulphonamide were included in the study. The list of antibiotics and their corresponding antibiotic concentration breakpoints suggested by CLSI are provided in Table S2 [55 ]. Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, and Pseudomonas aeruginosa ATCC 27853 (Oxoid company, Canada) were used as quality control (QC) strains.
5
Antibacterial Evaluation of PVA Films
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The antibacterial properties of the obtained PVA films loaded with plant extracts were evaluated by the Kirby–Bauer disk diffusion method [39 ]. Staphylococcus aureus ATCC 25923 (Gram-positive bacteria) (Thermo Fisher Scientific Inc., Dartford, UK) and Escherichia coli ATCC 25922 (Gram-negative bacteria) (Thermo Fisher Scientific Inc, Dartford, UK) were put in contact with the PVA film samples (6 mm). The protocol implied the preparation of a bacterial inoculum with 0.9% NaCl dilution and a turbidity of 0.5 on the McFarland scale (1.5 × 108 bacterial cells/mL) for 24 h of cultured cells. The bacterial cultures were incorporated in a sterile Mueller–Hinton (Oxoid), melted and cooled to 45 °C. The PVA films loaded with plant extracts and paper discs impregnated with extracts (80 µL) were placed at a relatively equal distance between them onto one Petri dish with Mueller–Hinton agar, inoculated with bacteria suspensions. The plates were prepared in duplicate and incubated at 37 °C for 24 h. After the incubation, the area of the microbial inhibition zone for each sample was measured.
6
Antibacterial Potential of Sage Extracts
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The antibacterial activity of Salvia officinalis extracts was assessed against Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, and Staphylococcus aureus ATCC 25923 (Thermo Fisher Scientific-Waltham, MA, USA).
7
GBS Identification Protocol for Clinical Samples
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The samples were processed as recommended by the American Society for Microbiology’s guidelines24 (link). Briefly, after incubating the swabs in Todd-Hewitt broth (Thermo Scientific™, Singapore) aerobically at 37 °C for 18–24 h, 10 μl of each broth was subcultured on Columbia agar plates with 5% sheep blood (Oxoid, Singapore). The plates were incubated for 24 h at 37 °C in 5% CO2. If GBS was not detected, the blood agar plate was incubated and examined after 48 h. All suspected GBS appeared as either beta-haemolytic or nonhemolytic, and Gram-positive cocci and catalase-negative cocci were taken for the CAMP (Christie–Atkins–Munch-Peterson) test. Streptococcus pyogenes ATCC 19615, Streptococcus agalactiae ATCC12386, and Staphylococcus aureus ATCC 25923 (Thermo Scientific™, Singapore) were the controls. All colonies that yielded a positive CAMP were considered GBS. Positive CAMP test results were confirmed using the Streptex™ Latex Agglutination test (Thermo Scientific™, Singapore) according to the manufacturer’s instructions. This latex agglutination test provides a complete solution for the isolation and differentiation of Lancefield Groups A, B, C, D, F, and G.
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