Tryptone soy broth (tsb)

Similarly, S. aureus was isolated on mannitol salt agar (Liofilchem, Italy), a selective and differential culture medium, by inoculation of 200 μL of each undiluted sample. Presumptive S. aureus isolates were purified on nutrient agar, Gram-stained, and tested for catalase and coagulase production. Confirmation of S. aureus identity was done using molecular methods. The genomic DNA of each presumptive S. aureus isolate was extracted using the simple boiling method. Pure colonies of S. aureus isolates were inoculated into 200 μL of tryptone soy broth (Merck, Darmstadt, Germany) and cultivated overnight at 37°C. The cells were harvested by centrifugation and suspended in 200 μL sterile physiological buffered saline. The suspension was heated in a water bath (Yamato Scientific, USA) at 100°C for 15 min and immediately chilled on ice. The boiled bacterial cells were centrifuged at full speed for 10 min in a microcentrifuge to separate the cell debris from the supernatant. The supernatant was stored at –20°C until used as DNA template.

Thirty-two UTIs E. coli isolates were collected from different human urine samples at Suez Canal University Hospital and gifted to Center for Microbiology and Phage Therapy at Zewail City of Science and Technology. In addition, ten E. coli strains from E. coli reference (ECOR) collection were used in this study. Bacterial isolates were identified and confirmed through culturing on differential selective Eosin Methylene Blue media (EMB-Oxoid, UK) and MacConkey agar (Oxoid, UK) prior to additional subcultures. Furthermore, the bacterial identity was validated and confirmed by PCR, in which QIAamp DNA Mini kit (Qiagen, Germany) was used for DNA extraction from bacterial isolates. Furthermore, size of the PCR product was determined by running it on an agarose gel (1.0% (w/v) agarose). Tryptone Soy Agar (TSB-Merck, Germany) was used in all study experiments. Tryptone Soy Broth (Merck, Germany) supplemented with 20–25% (w/v) glycerol was used to preserve all isolates at -80 °C until they were needed.

Ethanol, methanol, acetone, acetonitrile, Mueller Hinton broth (MHB), Mueller Hinton agar (MHA), Tryptone soy broth (TSB), Tryptone soy agar (TSA), Sabouraud dextrose broth (SDB), Sabouraud dextrose agar (SDA), Bovine serum albumin (BSA), Phosphate Buffer Saline (PBS), Triton-X 100, p-iodonitrotetrazolium chloride (INT), Thiazolyl Blue Tetrazolium Bromide (MTT), Amphotericin B, ciprofloxacin and Dimethyl sulphoxide (DMSO) were purchased from Sigma Aldrich, South Africa. Gentamicin was purchased from Virbac, New Zealand, Silica gel 60 from Merck, Germany, Minimal essential medium (MEM), Dulbecco’s Modified Eagle’s Medium (DMEM) and foetal calf serum (FCS) from Highveld Biological, South Africa. Cell lines C3A human hepatocyte (ATCC No. CRL-10741) and human colon (Caucasian colon adenocarcinoma (Caco2)) (ATCC HTB 37) were purchased from America Type Culture Collection, while Vero African green Monkey kidney cells was obtained from the collection of the Department of Veterinary Tropical Diseases, University of Pretoria.

Bacterial strains and plasmids used in this study are described in Tables S1 and S2, respectively. Oligonucleotides are listed in Table S3. P. aeruginosa strains (PAO1, PA14, and PAK) were grown in tryptone soy broth (Sigma), LB, or AB minimal medium supplemented with thiamine and glucose (ABTG) (15.1 mM (NH₄)₂SO₄, 33.7 mM Na₂HPO₄ × 2 H₂O, 22 mM KH₂PO₄, 50 mM NaCl, 1 mM MgCl₂ × 6 H₂O, 100 μM CaCl₂ × 2 H₂O, 1 μM FeCl₃ × 6 H₂O, 0.4 g of glucose per liter) and supplemented with appropriate antibiotics (carbenicillin 100 μg/ml, tetracycline 100 μg/ml or streptomycin 2 mg/ml) at 37 °C with agitation. P. putida (KT2440) was grown in either tryptone soy broth or LB medium at 30 °C and supplemented with appropriate antibiotic (streptomycin 2 mg/ml). Escherichia coli strains were grown in LB supplemented with antibiotics where appropriate (kanamycin 50 μg/ml, streptomycin 50 μg/ml, or gentamicin 50 μg/ml). IPTG was used, where stated, for P. aeruginosa at 250 μM and for E. coli at 500 μM final concentration. The E. coli strain DH5α was used as cloning host and BL21(DE3) was utilized for protein expression. P. aeruginosa chromosomal mutants were constructed as described (72 (link)). In brief, pKNG101 was transferred via three-partner conjugation using E. coli CC118 λpir as a donor and E. coli 1047/pRK2013 as a helper strain with counterselection by growth on 20% (w/v) sucrose.

HYDROGEL was supplied by POLISA Biopolímeros para a Saúde Ltd.a. Vancomycin, Müeller–Hinton agar (MHA), Müeller–Hinton broth (MHB), tryptone soy broth (TSB), glucose (D-(+)-Glucose), and crystal violet were purchased from Sigma-Aldrich (St. Louis, MO, USA). Methanol, glacial acetic acid, and all reagents were from Merck (Darmstadt, Alemanha).