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One millilitre of sample was transferred to Lysing Matrix E tubes and centrifuged at 16,000× g for 10 mins.
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One millilitre of honey sample was taken in a test tube and diluted to 10 ml by hot distilled water of 40 °C.
One millilitre of each sample at a concentration of 1 mg/ml were taken and diluted, to prevent quenching effects, with an appropriate volume of PBS.
One millilitre of the sample was diluted in 9 ml of Brain Heart infusion (BHI) (Oxoid, Basingstoke, England) and this suspension was serially diluted in BHI.
One millilitre of each water sample was pipetted into the sterile petri dish.
One millilitre of filtered sewage sample and 0.5 ml of K. pneumoniae 77 ESBL, grown overnight in Mueller Hinton Broth (MHB; bioMérieux Polska, Warsaw, Poland), were added to 10 ml of MHB and incubated at 37°C until complete lysis appeared (approximately 4 6 h).
Briefly, sediment sample taken from the same site was used as a growth medium in an inverted Tissue Culture Insert (TCI), and one millilitre of a 1 100 dilution sample was filtered on a Polycarbonate Membrane (PCM).
One millilitre of each collected PreservCyt sample was aliquoted with an individual sterile pipette for molecular study.
One millilitre of 80%% ethanol was added to each sample and the tubes were incubated for 90 min at 70 °C in a water bath.
One millilitre of CM was harvested on DIV4 and DIV8, on DIV12 cells and medium were harvested.
One millilitre of HPLC-grade water was then added, and samples were analyzed by ICP-AES (Perkin Elmer Optima 8300).
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