The minimum inhibitory concentration (MIC) and the minimum biocidal concentration (MBC) were determined by serial twofold dilutions. Test strains used in the study were S. aureus, E. coli, P. aeruginosa, and C. albicans from MicroBioLogics, Inc. (St. Cloud, USA). The MIC of the preparations was determined on liquid media: Mueller-Hinton broth (Merck, Germany) for bacteria and Sabouraud broth (BTL, Poland) for C. albicans. 0.1 ml of inoculum (24-h strains of 0.5 McFarland density) was introduced into prepared tubes with the medium and tested preparation. The control was a tube of Mueller-Hinton broth (10 ml) without any preparation (blank test). Samples were incubated at 37 °C for 24 h (bacteria) and 72 h (yeast fungi). The MIC value was taken as the concentration of the preparation that completely inhibited the growth of microorganisms (no turbidity in the tube). In order to determine the MBC from cultures of bacteria and fungi considered negative, cultures were performed on solid Triptic Soy Agar and Sabouraud medium (BTL, Poland). After 24 h and 72 h (bacteria and fungi) of incubation at 37 °C, a culture determination (macroscopic evaluation) was performed. MBC was defined as the lowest concentration of the formulation resulting in complete inhibition of bacterial/fungal growth.
S aureus
Manufactured by Microbiologics
Sourced in United States
The S. aureus product is a strain of Staphylococcus aureus bacteria. It is a gram-positive, spherical-shaped bacterium that can be used for various microbiological applications. The core function of this product is to serve as a reference strain for identification, detection, and quality control purposes in a laboratory setting.
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Lab products found in correlation
P aeruginosa, by Microbiologics (3 mentions)
E coli, by Microbiologics (2 mentions)
Mueller hinton broth (mhb), by Merck Group (1 mentions)
C albicans, by Microbiologics (1 mentions)
Atcc 25923, by Microbiologics (1 mentions)
S aureus atcc 25923, by Microbiologics (1 mentions)
K pneumoniae, by Microbiologics (1 mentions)
S epidermidis, by Microbiologics (1 mentions)
Salmonella typhimurium, by Microbiologics (1 mentions)
Escherichia coli, by Microbiologics (1 mentions)
Trypticase soy agar, by Merck Group (1 mentions)
Trypticase soy broth (tsb), by Merck Group (1 mentions)
Ma 420, by Marconi (1 mentions)
6 protocols using s aureus
1
Antimicrobial Susceptibility Testing Protocol
2
Antimicrobial Evaluation of β-Cyclodextrin Derivatives
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β-CD (Wako 1st Grade) and 2-HP-β-CD were obtained from FUJIFILM Wako Chemicals (Osaka, Japan) and Tokyo chemical industry Co., Ltd. (Tokyo, Japan), respectively. HEO, extracted from the xylem of Hinoki, was bought from INSCENT CO. (Nagano, Japan). Ethanol (guaranteed reagent, 99.5%) was purchased from FUJIFILM Wako Chemicals. All other chemicals and solvents were analytical grade and were used directly without further purification.
S. aureus, derived from ATCC 25923, was bought from Microbiologics, Inc. (Saint Cloud, MN, USA). Tryptic soy agar (TSA) and Tryptic soy broth (TSB) were purchased from Geno Technology Inc. (St. Louis, MO, USA).
S. aureus, derived from ATCC 25923, was bought from Microbiologics, Inc. (Saint Cloud, MN, USA). Tryptic soy agar (TSA) and Tryptic soy broth (TSB) were purchased from Geno Technology Inc. (St. Louis, MO, USA).
3
Antibacterial Potential of Thiosemicarbazides
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Minimal inhibitory concentration (MIC) of each antibacterial agents was determined by standardized broth microdilution method with inoculum of 5 × 105 CFU/mL and incubated for 24 h at 35 °C according to the recommendation of CLSI (Clinical Laboratory Standards Institute) [35 ]. The antibacterial activity of the thiosemicarbazides 1 –9 and antibiotics (amoxicillin, gentamicin, levofloxacin, linezolid, and vancomycin) was tested on the Gram-positive strains (Microbiologics, St. Cloud, MN, USA) (S. aureus ATCC 25923, S. aureus ATCC 6538, S. epidermidis ATCC 12228, B. subtilis ATCC 6633, B. cereus ATCC 10876, M. luteus ATCC 10240), the Gram-negative strains (Microbiologics, St. Cloud, MN, USA) (E. coli ATCC 25922, K. pneumoniae ATCC 13883, P. mirabilis ATCC 12453, P. aeruginosa ATCC 9027), and clinical isolates of S. aureus (Medical University of Lublin, Lublin, Poland) (MSSA-1, MSSA-2, MSSA-3, MSSA-4, MRSA-1). Similarly to previous experiments with the thiosemicarbazides 1 –5 , cefuroxime was used as positive control antibiotic [23 (link)], whereas sterile distilled water served as negative controls.
4
Microbial Strain Preparation and Storage
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K.
pneumoniae (VTCC 12273) and Pseudomonas
aeruginosa (VTCC 12018) were provided by the Institute
of Microbiology and Biotechnology, Vietnam National University. Escherichia coli (ATCC 25922), Clostridium
perfringens (ATCC 12915), S. aureus (ATCC 11632), and Salmonella typhimurium (ATCC 14028) were purchased from Kwik-Stik, Microbiologics Inc.,
USA. The strains were grown at 37 °C in BHI broth until an optical
density (OD) at 600 nm of 0.8 (108 CFU/mL) was reached. P. aeruginosa, E. coli, C. perfringens, S.
aureus, and S. typhimurium were used as negative controls. K. pneumoniae and the negative controls were cultured in BHI agar to determine
the serial dilution and calculate CFUs/mL. The bacteria were cryopreserved
in glycerol and stored at −20 °C.
pneumoniae (VTCC 12273) and Pseudomonas
aeruginosa (VTCC 12018) were provided by the Institute
of Microbiology and Biotechnology, Vietnam National University. Escherichia coli (ATCC 25922), Clostridium
perfringens (ATCC 12915), S. aureus (ATCC 11632), and Salmonella typhimurium (ATCC 14028) were purchased from Kwik-Stik, Microbiologics Inc.,
USA. The strains were grown at 37 °C in BHI broth until an optical
density (OD) at 600 nm of 0.8 (108 CFU/mL) was reached. P. aeruginosa, E. coli, C. perfringens, S.
aureus, and S. typhimurium were used as negative controls. K. pneumoniae and the negative controls were cultured in BHI agar to determine
the serial dilution and calculate CFUs/mL. The bacteria were cryopreserved
in glycerol and stored at −20 °C.
5
Propagation of Common Bacterial Strains
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The stocks of Gram-positive (S. aureus, B. cereus) and Gram-negative (P. aeruginosa, E. coli) strains used in this study were obtained from Microbiologics (Minnesota, USA) and subsequently propagated on Mueller-Hinton broth (MH) for 24 h at 37°C before use.
6
Characterization of S. aureus and P. aeruginosa
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For the study, standard strains of S. aureus American Type Culture Collection (ATCC) 6538 and P. aeruginosa ATCC 15442 were used, acquired from Microbiologics® and provided by the company Golden Technology®. These were maintained in Trypticase Soy Broth (TSB, Merck, Darmstadt, Germany) with 20% glycerol between −18 and −20 °C, subcultured in Trypticase Soy Agar (TSA, Merck, Darmstadt, Germany), using up to the fifth passage at most. Bacteria were cultured 24 h before the experiment was carried out. Sequentially, an isolated colony of the microorganism was inoculated into 10 mL of Brain Heart Infusion Broth (BHI, Merck, Darmstadt, Germany) and incubated for 24 h at 37 °C under agitation in a shaker (Marconi, MA420) at 200 rotations per minute (rpm). The purity and viability of the cultures were also tested, according to strict laboratory biosafety standards.
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