BacterPlate™ Mueller Hinton Agar, Glucose-Methylene Blue

• Prepare inoculum by picking five distinct colonies (~1mm) from a 24-hour-old culture grown on BacterPlate™ Sabouraud Dextrose Agar and incubated at 35 ± 2°C. Suspend colonies in 5ml of sterile 0.85% saline.
• Vortex the suspension and adjust turbidity to yield 1×10⁶ – 5×10⁶ CFU/ mL (0.5 McFarland standard).
• Prepare sterile BacterPlate™ Mueller Hinton Agar, Glucose-Methylene Blue: Allow to warm to room temperature before inoculation, dry the plates in an incubator with the covers partially removed.
• Dip a sterile, non-toxic cotton swab into the standardized inoculum. Press the swab against the inner tube wall to remove excess fluid. Streak the agar surface three times, turning the plate 60° between streaks. Allow the inoculum to dry for 5 – 15 minutes with the lid in place.
• Apply antifungal discs aseptically, ensuring centers are at least 24 mm apart. Invert the plates and incubate at 35 ± 2°C within 15 minutes of disc application.
• Examine plates after 20 – 24 hours of incubation. Ensure zones of inhibition are uniformly circular and a semi-confluent lawn of growth is present. If insufficient growth is observed, read after 48 hours

RESULTS

• Measure the inhibition zones.
• Refer to the table for interpreting inhibition zones furnished by the suppliers of antibiotic disks in order to establish the correlation between the inhibition zone and the minimal inhibitory concentration (M.I.C.)