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.
P aeruginosa
Manufactured by Microbiologics
Sourced in United States
P. aeruginosa is a lyophilized microorganism product used for quality control and proficiency testing purposes in microbiology laboratories. The product contains Pseudomonas aeruginosa bacteria, a common environmental microorganism.
Automatically generated - may contain errors
Lab products found in correlation
S aureus, by Microbiologics (3 mentions)
C albicans, by Microbiologics (2 mentions)
K pneumoniae, by Microbiologics (2 mentions)
E faecalis, by Merck Group (1 mentions)
K pneumoniae, by Merck Group (1 mentions)
B subtilis, by Merck Group (1 mentions)
S aureus, by Merck Group (1 mentions)
P aeruginosa, by Merck Group (1 mentions)
S aureus, by American Type Culture Collection (1 mentions)
E faecalis, by American Type Culture Collection (1 mentions)
S epidermidis, by Microbiologics (1 mentions)
Mueller hinton broth (mhb), by Merck Group (1 mentions)
4 protocols using p aeruginosa
1
Propagation of Common Bacterial Strains
2
Standardized Microbial Inoculum Preparation
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S. aureus (American Type Culture Collection, ATCC 6538), E. faecalis (ATCC 29212), B. subtilis spizizenii (ATCC 6633), E. cloacae (ATCC 35030), K. pneumoniae (ATCC 10031), A. brasiliensis (ATCC 16404), P. aeruginosa (ATCC 9027) and C. albicans (ATCC 10231) were obtained from Microbiologics (Minnesota, USA). F. solani (Deutsche Sammlung von Mikroorganismen, DSM 1164) was supplied by Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Germany.
Inocula of S. aureus, E. faecalis, B. subtilis, E. cloacae, A. brasiliensis, P. aeruginosa, C. albicans, K. pneumoniae and F. solani were prepared from lyophilised pellets according to the manufacturer’s protocols or from fresh culture according to McFarland standards to obtain 1.0×107 colony forming units (CFU/mL in physiological solution (0.9% (w/v) NaCl); Merck-Sigma-Aldrich, Darmstadt, Germany.
Tryptone soya broth (20 mL; Bioculturalab, Italy) was used as growth control for C. albicans, A. brasiliensis and F. solani, and thioglycolate medium (20 mL; Bioculturalab) was used as growth control for S. aureus, B. subtilis, P. aeruginosa and K. pneumoniae. Nutrient broth (Nutrient Broth, 20 mL; BioLife, Italy) medium was used as growth control for E. faecalis and E. cloacae.
Inocula of S. aureus, E. faecalis, B. subtilis, E. cloacae, A. brasiliensis, P. aeruginosa, C. albicans, K. pneumoniae and F. solani were prepared from lyophilised pellets according to the manufacturer’s protocols or from fresh culture according to McFarland standards to obtain 1.0×107 colony forming units (CFU/mL in physiological solution (0.9% (w/v) NaCl); Merck-Sigma-Aldrich, Darmstadt, Germany.
Tryptone soya broth (20 mL; Bioculturalab, Italy) was used as growth control for C. albicans, A. brasiliensis and F. solani, and thioglycolate medium (20 mL; Bioculturalab) was used as growth control for S. aureus, B. subtilis, P. aeruginosa and K. pneumoniae. Nutrient broth (Nutrient Broth, 20 mL; BioLife, Italy) medium was used as growth control for E. faecalis and E. cloacae.
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
Antimicrobial Susceptibility Testing Protocol
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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.
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