Glucose bromocresol purple agar bcp

For bacteria isolation, serial dilution method (10^–1–10^–3) in sterile peptone water (Sigma Aldrich, Steinheim, Germany) was applied. After defrosting, samples were thoroughly vortexed and then 0.5 mL was transferred into the test tube containing 4.5 mL of sterile peptone water (Sigma Aldrich, Germany) and again vortexed (first dilution—10^–1). 100 μL of each dilution was plated onto 5 different culture media previously selected as the most useful in DFI bacteria recovery [8 (link)]: Tryptic Soy Agar (TSA; Sigma Aldrich, Steinheim, Germany), Columbia Blood Agar (BLA; Oxoid, Basingstoke, Great Britain), CHROMagar Orientation (CHRA; GRASO Biotech, Starogard Gdański, Poland), Glucose Bromocresol Purple Agar (BCP; Sigma Aldrich, Steinheim, Germany), and Vancomycin Resistant Enterococci Agar (VRE; Oxoid, Basingstoke, Great Britain). All media were in the form of ready-to-use powders except for BLA, which was prepared by adding defibrinated sheep blood (GRASO Biotech, Starogard Gdański, Poland) to the sterilized and dissolved Colombia blood agar base to the final concentration 5% (v/v). Bacterial cultures were incubated at 37 °C for 24 h and then single colonies characterized by different morphological features were selected to obtain pure cultures using the streak plate method on the same media (incubation at 37 °C for 18–24 h).

After defrosting, samples were thoroughly vortexed and a series of 10-fold serial dilution (10⁻¹ to 10⁻³) were prepared from them. For this purpose, 0.5 mL of the clinical sample was added to a test tube containing 4.5 mL of sterile peptone water (Sigma Aldrich, Steinhelm, Germany) and vortexed (first dilution—10⁻¹). 100 μL of each dilution was plated onto 10 different types of culture media: Tryptic Soy Agar (TSA; Sigma Aldrich, Germany), Mueller Hinton Agar (MHA; Sigma Aldrich, Germany), Columbia Agar Base (COL; Oxoid, UK), Columbia Blood Agar (BLA; Oxoid, UK), Brain Heart Infusion Agar (BHI; Sigma Aldrich, Germany), CHROMagar Orientation (CHRA; GRASO Biotech, Poland), Azide Blood Agar (AZI; Oxoid, UK), Glucose Bromocresol Purple Agar (BCP; Sigma Aldrich, Germany), Mannitol Salt Agar (MAN; Oxoid, UK), Vancomycin-Resistant Enterococi Agar (VRE; Oxoid, UK). All media were in the form of ready-to-use powders except for BLA, which was prepared by adding defibrinated sheep blood (GRASO Biotech, Poland) to the sterilized and dissolved Colombia blood agar base to the final concentration 5% (v/v). After incubation at 37 °C for 18–24 h on the basis of morphological differences, single colonies of bacteria were selected, from which reduction cultures were prepared on the same media in order to obtain pure cultures (incubation at 37 °C for 18–24h).