Muller hinton agar

Antimicrobial activity was assessed by viable cell count assay, using protocols already reported in literature [24] , with some modifications.
Briefly, liquid cultures of selected microorganisms were grown overnight at 37 °C in Mueller-Hinton broth, and the bacterial concentration adjusted to approximately 10 +6 colony forming units (CFU) per mL. 8.5 g•L -1 sodium chloride and 1.0 g•L -1 peptone water solutions were used for dilution.
1.3 cm diameter film discs (CS film control and CS-FeHAp film) were sterilised under UV light for 10 min on each side. The film discs were placed in sterile tubes, and 200 μL of liquid inoculum were placed over each disc. The films were left in contact with the inoculum for 2, 4, and 8 h at 37 °C. At each sampling time, 1.8 mL of dilution solution was added to each tube, and successive dilutions were prepared. Aliquots (20 μL) of each diluted solution were planted on Muller-Hinton agar (Biokar Diagnostics, France) and, after incubation at 37 °C, colonies were counted. Each experiment was performed in triplicate, with statistical analysis performed as described in Section 2.3.8.
The tested microorganisms were a Gram-negative bacteria, Escherichia coli (ATCC 25922), a Gram-positive bacteria, methicillinresistant Staphyloccocus aureus (MRSA, CCUG 60578), and yeast, Candida albicans (CBQF isolate).