Pseudomonas aeruginosa atcc 9027

Twelve L. paracasei strains, namely, KCTC 3169, KCTC 13090, KCTC 3165, KCTC 3189, KCTC 5546, KCTC 3510, KCTC 5058, KCTC 3074, KCCM 40995, KCCM 42830, KCCM 32822, and KCCM 41276 were purchased from the Korean Culture Collection for Type Cultures (Daejeon, Korea) and Korean Culture Center of Microorganisms (Seoul, Korea). L. paracasei KB28, L. paracasei DLP 1354, Lactobacillus gasseri DLP1202, Bifidiobacterium bifidum DLP1224, B. animalis DLP1267, B. faecale DLP1470, Pseudomonas aeruginosa ATCC 9027, Bacillus subtilis ATCC 6633, Escherichia coli ATCC 8739, Staphylococcus aureus ATCC 6538, and Clostridium difficile, Aspergillus brasiliensis ATCC 16404 were provided by DONG-A PHARM (Seoul, Korea). The strains were grown in De Man, Rogosa, and Sharp (MRS, Oxoid, Hampshire, UK) at 37 °C for 24 h under anaerobic condition. The strains were subcultured weekly on MRS agar and stored at 4 °C.

Tested bacteria, including 2 gram-positive (gram (+)), such as Bacillus subtilis ATCC 6633 (B. sub), Staphylococcus aureus ATCC 25923 (S. aureus), and 4 gram-negative (gram (-)), such as Escherichia coli (E. coli) ATCC 25922, E. coli ATCC 85922, Pseudomonas aeruginosa ATCC 9027, Salmonella typhimurium ATCC 13311, were purchased from the American Type Culture Collection(ATCC, Rockville, MD, USA).

Two strains of pathogenic bacteria, Pseudomonas aeruginosa ATCC 9027, Staphylococcus aureus CIP 65.8^T, which are responsible for a large number of postoperative infections, were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and the Culture Collection of the Institute Pasteur (CIP, Paris, France) [1 (link), 27 (link)]. The selected strains are representatives of two large prokaryotic lineages, namely Gram-negative and Gram-positive bacteria, whose responses on nanostructured surfaces will be typical for taxonomically related bacterial taxa. Prior to each experiment, bacterial cultures were refreshed on nutrient agar from stocks (BD, USA). Fresh bacterial suspensions were grown overnight at 37 °C in 5 mL of nutrient broth (BD, USA). Bacterial cells were collected at the logarithmic stage of growth (data not shown), and the density of bacterial suspensions was adjusted to OD₆₀₀ = 0.1.

The Acinetobacter baumannii ATCC BAA-1605, Candida albicans ATCC 10231, Enterococcus faecium ATCC 700221, Klebsiella aerogenes ATCC 13048 (previously known as Enterobacter aerogenes), Klebsiella pneumoniae ATCC 700603, Pseudomonas aeruginosa ATCC 9027, Staphylococcus aureus ATCC 25923 and MRSA ATCC 33591, were acquired from the American Type Culture Collection (ATCC). Before running the tests, beads with cryo-protected microorganisms were transferred into fresh MHB (BioMaxima, Lublin, Poland) for bacteria or RPMI-1640 for fungi and incubated for 24 h at 37 °C. Then, the culture was seeded on the Mueller-Hinton agar (MHA) plates (BioMaxima, Lublin, Poland) for bacteria or Sabouraud dextrose agar (SDA) plates (BioMaxima, Lublin, Poland) for fungi and incubated as just mentioned. Cell densities for all assays were adjusted spectrophotometrically (Multiskan™ GO Microplate Spectrophotometer, Thermo Fisher Scientific, Vantaa, Finland) at 600 nm for bacteria and at 530 nm for fungi.

Staphylococcus aureus ATCC25923, Pseudomonas aeruginosa ATCC9027, Escherichia coli ATCC25922, and Lactobacillus sp. ATCC7469 were purchased from American Type Culture Collection (ATCC) and preserved at -70°C in our laboratory. Before use, these strains were activated in LB liquid medium to exponential phase at 37°C and 220 rpm.

Pseudomonas aeruginosa ATCC 9027, P. aeruginosa PAO1 ATCC 15692, Escherichia coli O157:H7 ATCC 51659, and Staphylococcus aureus ATCC 25923 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). F. novicida U112 NR-13 was obtained from BEI Resources (Manassas, VA, USA). P. aeruginosa PAO1 cells that constitutively expressed GFP (PAO1 pTDK-GFP) and S. aureus SH1000 that expressed RFP (SH1000 pAH9-RFP) were kindly provided by Dr. Weibel^{57 (link)} and Dr. Boles,^{58 (link)} respectively. DRGN-1 and VK25 were synthesized by the conventional solid-phase method by ChinaPeptides (Suzhou, China). The crude compounds were purified to chromatographic homogeneity in the range of >95% by using reversed-phase high-performance liquid chromatography with a mass spectrometer by the company. LL-37 was purchased from AnaSpec (Fremont, CA, USA). All peptides were reconstituted in a buffer consisting of 20 mg bovine serum albumin and 10 µl acetic acid in 50 ml PBS.^{59 (link)} Table 2 summarizes characteristics of peptides used in this study.

A reference strain of Pseudomonas aeruginosa (ATCC 9027) was purchased from the American Type Culture Collection (ATCC). The strain was streaked on Luria-Bertani (LB) agar and incubated for 24 hrs at 37 °C. After incubation, the strain was inoculated into sterile LB broth and incubated at 37 °C for 12 hrs. The bacterial suspension was then adjusted to be 0.5 McFarland. Then, the strain was stored in Luria-Bertani (LB) broth containing 20% (v/v) glycerol at −80 °C [19] –[21] (link).