Pseudomonas aeruginosa atcc 27853

Manufactured by American Type Culture Collection

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Pseudomonas aeruginosa ATCC 27853 is a well-characterized bacterial strain belonging to the Pseudomonas genus. This strain is commonly used as a quality control organism in various microbiological testing procedures.

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55 protocols using pseudomonas aeruginosa atcc 27853

Antimicrobial Activity of 3D Matrix

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The antimicrobial activity of the 3D anti-infective regenerative matrix was tested using Gram positive (Staphylococcus aureus ATCC^® 25923) and Gram negative (Pseudomonas aeruginosa ATCC^® 27853) bacterial strains purchased from American Type Culture Collection (ATCC). Glycerol stocks were streaked on LB agar plates and after 24h of incubation bacterial suspensions of 0.5 McFarland density corresponding to a 1–3 × 10⁸ CFU (colony forming units)/mL density were obtained. This suspension was used to uniformly streak the whole surface of fresh LB agar plates. After inoculation the plates were separated into two sets (labeled T₀ and T₁). For the T₀ set, a 5 mm × 5 mm piece of each tested material was added on the inoculated plate using sterile tweezers. For the T₁ set, the bacteria were allowed to grow for 6 h at 37 °C and, after this time, lapse pieces of the tested materials were added on the plate. Both sets have been incubated for another 24 h, and then plates were analyzed and the growth inhibition zones were measured and converted in percentages. The experiment was performed in triplicate and repeated on at least three separate occasions.

Ion A., Andronescu E., Rădulescu D., Rădulescu M., Iordache F., Vasile B.Ș., Surdu A.V., Albu M.G., Maniu H., Chifiriuc M.C., Grumezescu A.M, & Holban A.M. (2016). Biocompatible 3D Matrix with Antimicrobial Properties. Molecules, 21(1), 115.

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Antibacterial and Cytotoxicity Evaluation

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Lipopolysaccharide (LPS; from Pseudomonas aeruginosa), Lipoteichoic acid (LTA; from Staphylococcus aureus), 3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI), n-phenyl-1-naphthylamine (NPN), HEPES, and 3,3′-dipropylthiadicarbocyanine iodide (DisC₃-5) were obtained from Sigma-Aldrich (St Louis, MO, USA).
Pseudomonas aeruginosa ATCC 27853 and Staphylococcus aureus ATCC 25923 were obtained from the ATCC (American Type Culture Collection, Manassas, VA, USA), and Acinetobacter baumannii KCTC 2508 and Bacillus subtilis KCTC 2217 were obtained from the KCTC (Korean Collection for Type Cultures, Jeongeup-si, Jeollabuk-do, Korea). Human skin epithelial cells (HaCaT cells) were obtained from ATCC.

Kwon J.Y., Kim M.K., Mereuta L., Seo C.H., Luchian T, & Park Y. (2019). Mechanism of action of antimicrobial peptide P5 truncations against Pseudomonas aeruginosa and Staphylococcus aureus. AMB Express, 9, 122.

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Antibacterial Evaluation of Compounds

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The antibacterial activity of each compound was evaluated using six bacterial strains: three strains of Gram-positive (Staphylococcus aureus ATCC 292313, Bacillus subtilis ATCC 6633, and Enterococcus faecalis ATCC 29212) and three strains of Gram-negative (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, and Klebsiella pneumoniae ATCC 700603) bacteria. The bacterial strains were purchased from American Type Culture Collection (ATCC) (Manassas, VA, USA).

Kania A., Tejchman W., Pawlak A.M., Mokrzyński K., Różanowski B., Musielak B.M, & Greczek-Stachura M. (2022). Preliminary Studies of Antimicrobial Activity of New Synthesized Hybrids of 2-Thiohydantoin and 2-Quinolone Derivatives Activated with Blue Light. Molecules, 27(3), 1069.

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Isolation and Characterization of Bacteria from Environmental Samples

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The natural specimens consisting of 100 mg of Bangkhuntien's mangrove forest soil, Nam Nao National Park's soil, and Yaowarat Chinese market herbs were mixed with sterile distilled water to final volume of 10 mL. For Rayong's seawater, 10 mL was collected and diluted 1,000 times with NSS before use. After homogenization, 100 µL of the aqueous solutions was plated on agar plate of CDA, MEA, MRS, NA, PCA, PDA, SDA, and TSA (Difco, USA). We incubated the plate 3 days and then transferred single colony into new NA agar plate. During doing bacteria culture, bacterial Gram of all strains was identified by standard Gram staining. In this study, we used 4 importance human pathogens as standards for antibacterial assay, namely, Bacillus cereus ATCC14579, Escherichia coli ATCC25922, Pseudomonas aeruginosa ATCC27853, and Staphylococcus aureus ATCC25923 (ATCC, USA). In order to maintain all the isolated and standard bacterial strains, we kept their 3-day NB cultures with 25% glycerol and restocked them every 6 months at −80°C. All bacteria strains' genomic DNA was prepared using Presto™ Mini gDNA Bacteria Kit following the manufacturer protocol (Geneaid, Taiwan).

Phonghanpot S, & Jarintanan F. (2022). Secondary Metabolism Gene Diversity and Cocultivation toward Isolation and Identification of Potent Bioactive Compounds Producing Bacterial Strains from Thailand's Natural Resources. Scientifica, 2022, 2827831.

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Antimicrobial Screening of P. oxalicum

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The fungal strain
was obtained from
the national microbial repository, that is, P. oxalicum (VRE-2) was procured from Col. Sir R. N. Chopra, Microbial Resource
Center Jammu (MRCJ), India, under accession number MRCJ-1. The strain
was cultured in PDB with constant shaking at 150 rpm for 7 days at
28 °C and on PDA in Petri dishes for 3–5 days at 28 °C. Staphylococcus aureus ATCC 25923, B. subtilis ATCC 6633, E. coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, one yeast, Candida albicans ATCC 24433, and Aspergillus niger ATCC 16404 used for antimicrobial activity screening were procured
from American Type Culture Collection (ATCC).

Abrol V., Kushwaha M., Arora D., Mallubhotla S, & Jaglan S. (2021). Mutation, Chemoprofiling, Dereplication, and Isolation of Natural Products from Penicillium oxalicum. ACS Omega, 6(25), 16266-16272.

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Synthesis and Antimicrobial Evaluation of AgNPs

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P. rhoeas leaves obtained from the southeast of Turkey (Kilis-Alatepe, 79002) were utilized in the reduction reactions of AgNO₃ salt. P. rhoeas GPS location: 36°47′13″ N, 37°11′57″ D. 36°47′13″ N, and 37°11′57″ D on the GPS.
The antibiotics used in the study (colistin for Gram-positive, fluconazole for yeast, and vancomycin for Gram-negative) and AgNO₃ (99.8 purity) were bought from Sigma-Aldrich (Darmstadt, Germany). Pathogenic Staphylococcus aureus (ATCC^® 29213™), Pseudomonas aeruginosa (ATCC^® 27853™), Bacillus subtilis (ATCC^® 11774™), Escherichia coli (ATCC^® 25922™), and Candida albicans (ATCC^® 10231™) were used to reckon the growth suppressive impact of generated AgNPs.

İpek P., Yıldız R., Baran M.F., Hatipoğlu A., Baran A., Sufianov A, & Beylerli O. (2023). Green Synthesis of Silver Nanoparticles Derived from Papaver rhoeas L. Leaf Extract: Cytotoxic and Antimicrobial Properties. Molecules, 28(17), 6424.

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Polymer-Antibiotic Nanocarrier Fabrication

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Poly(DL-lactic-co-glycolic acid) was obtained from Lakeshore Biomaterials (Birmingham, AL) and had a copolymer ratio of 50:50, a weight average molecular weight of 36 kDa, and a number average molecular weight of 21 kDa. Poly(vinyl) alcohol (PVA), Mueller-Hinton broth, ciprofloxacin, colistin sulfate, doxycycline hyclate, and vancomycin hydrochloride were obtained from Sigma Aldrich (St Louis, MO). Cefazolin sodium and clindamycin hydrochloride were obtained from Fisher Scientific (Waltham, MA). Properties of these antibiotics are detailed in Table 1. Colistin (polymyxin E) has two HPLC retention times due to this drug being a mixture of two drugs, colistin A and colistin B (Table 1). Experimental values for logP and logS were used when available, otherwise predicted values were calculated using ALOGPS (VCCLabs) [20 (link),21 (link)]. The charge of antibiotics at pH 6.4–7.4 is shown, which encompasses the pH of aqueous phases during fabrication and the pH of the release medium. ciprofloxacin was acidified with 5N HCl in order to create the water-soluble hydrochloride salt of ciprofloxacin. Bacterial strains Staphylococcus aureus (ATCC 29213), Escherichia coli (ATCC 25922), and Pseudomonas aeruginosa (ATCC 27853) were obtained from ATCC (Manassas, VA).

Shah S.R., Henslee A.M., Spicer P.P., Yokota S., Petrichenko S., Allahabadi S., Bennett G.N., Wong M.E., Kasper F.K, & Mikos A.G. (2014). Effects of Antibiotic Physicochemical Properties on their Release Kinetics from Biodegradable Polymer Microparticles. Pharmaceutical research, 31(12), 3379-3389.

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Antimicrobial Assessment of Metal Compounds

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LB media, zinc gluconate (TCI chemicals Pvt. Ltd, India), ferrous chloride (Sigma-Aldrich) were procured from the respective commercial sources. Staphylococcus aureus-ATCC 25923;
Escherichia coli-ATCC 25922; Pseudomonas aeruginosa-ATCC 27853 and Klebsiella pneumoniae-ATCC 700603 were purchased from ATCC, USA. Clinical specimens were obtained from PGIMER, Chandigarh after receiving the requisite ethics committee approval.

Kaushik S., Thomas J., Panwar V., Murugesan P., Chopra V., Salaria N., Singh R., Roy H.S., Kumar R., Gautam V., & Ghosh D. (2021). A Drug-free, Self-destruction Strategy to Combat Bacterial Infections by Using the Intrinsically formed Magnetic Nanoparticles in Bacterial Pathogens.

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Antibiotic Susceptibility Testing of Bacterial Strains

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Escherichia coli ATCC 25922, Enterobacter cloacae ATCC 13047, Klebsiella pneumoniae ATCC 13883, Pseudomonas aeruginosa ATCC 27853, and Acinetobacter baumannii 17978 were obtained from the American Type Culture Collection (Manassas, VA). Strains were stored at −80°C in tryptic soy broth (BD Diagnostics, Franklin Lakes, NJ) containing 50% glycerol (Sigma-Aldrich, St. Louis, MO).
Meropenem was from Ark Pharm (Arlington Heights, IL), cefepime was from Chem Impex (Wood Dale, IL), gentamicin was from Alfa Aesar (Haverhill, MA), and ciprofloxacin was from US Biological (Salem, MA). Antibiotic stock solutions were prepared in water for manual dilution-based testing or in water containing 0.3% polysorbate-20 as required for liquid handling by the HP D300 digital dispenser (HP Inc., Palo Alto, CA).⁹ We previously determined through extensive analysis that polysorbate-20 at the concentrations used in assay wells has no effect on MIC determinations for all antibiotics examined.^{5 (link), 10 (link)}

Smith K.P., Richmond D.L., Brennan-Krohn T., Elliott H.L, & Kirby J.E. (2017). Development of MAST: a Microscopy-Based Antimicrobial Susceptibility Testing Platform. SLAS technology, 22(6), 662-674.

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Antibiotic Resistance Profiling of Clinical Isolates

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One hundred and twenty (120) Gram-positive bacteria, including 20 methicillin-resistant Staphylococcus epidermidis (MRSE), 20 methicillin-sensitive Staphylococcus epidermidis (MSSE), 20 methicillin-resistant Staphylococcus aureus (MRSA), 20 methicillin-sensitive Staphylococcus aureus (MSSA), 20 Pyogenic streptococcus, 10 Entercoccus faecalis, 10 Enterococcus faecium and 380 gram-positive bacteria including 85 Escherichia coli, 80 Klebsiella pneumoniae, 50 Enterobacter cloacae, 15 Proteus mirabilis, 30 Enterobacter aerogenes, 15 Haemophilus influenzae, 25 Citrobacter freundii, 20 Serratia marcescens, 25 Pseudomonas aeruginosa, 15 Stenotrophomonas maltophilia, and 20 Acinetobacter baumannii were clinical isolates collected from hospitals in Beijing region during 2013–2014. Quality control bacteria such as Staphylococcus aureus ATCC25923, ATCC29213, Escherichia coli ATCC25922, ATCC35218, Pseudomonas aeruginosa ATCC27853, Klebsiella pneumoniae ATCC 10031, Entercoccus faecalis ATCC29212 were purchased from American Type Culture Collection center (ATCC, Manassas, VA, USA). β-Lactams in all the strains were determined by using the filter paper method and validated by the instructions of the American Clinical and Laboratory Standards Institute (CLSI, Wayne, PA, USA).

Yao Q., Wang J., Cui T., Yang Z., Su M., Zhao P., Yan H., Zhan Y, & Yang H. (2016). Antibacterial Properties of Tebipenem Pivoxil Tablet, a New Oral Carbapenem Preparation against a Variety of Pathogenic Bacteria in Vitro and in Vivo. Molecules, 21(1), 62.

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