Bacteria utilised in this study were as previously described [15]. Namely, neuropathogenic Escherichia coli K1 (a cerebrospinal fluid isolate from a meningitis patient; O18:K1:H7) (MTCC 710859) was cultivated. Several colonies of E. coli K1 were suspended in sterile nutrient broth. Following 10 h the optical density (OD) was adjusted to 595 nm to obtain a bacterial concentration of approximately 108 colony-forming units (CFU)/ml [45]. Ten microliters of E. coli K1 suspension were added to Phosphate Buffer Saline (PBS) containing 0.5 μg/ml AgNPs, or 0.5 μg/ml AgNPs-HDN. PBS alone was utilised as a control. Cultures were then treated with the aforementioned, in duplicates at various time intervals (0, 10, 20, 30, 40, 50, 60,70, 80, 90, 100, 110 and 120 mins). Following treatment, 10 μL of each aliquot were serially diluted and plated on nutrient agar in duplicates, followed by incubation at 37 °C for 16 h before bacterial colonies were enumerated.

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