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BacterPlate™ Mueller Hinton Agar, Glucose-Methylene Blue
- Ready-to-use medium on 90mm plates for testing susceptibility with antifungal disc diffusion of yeasts.
- Code: 05033
- Packaging: 10 plates/ box or as per customer request.
- Storage Conditions: 2–8°C.
- Transportation: Ambient temperature.
- Shelf Life: 03 months from the date of manufacture.
INTENDED USE
- BacterPlate™ Mueller Hinton Agar, Glucose-Methylene Blue is a specialized medium recommended for antifungal disk diffusion susceptibility testing of yeast species.
- The packaging with semi-permeable Cellophane film helps balance the humidity of the environment during storage.
PRINCIPLES
- Beef infusion and casein acid hydrolysate provide nitrogenous compounds, carbon, sulphur and other essential nutrients. Starch acts as a protective colloid against toxic substances present in the medium. Starch hydrolysis yields dextrose, which serves as a source of energy. Glucose serves as an energy source for fungal cultures while Methylene blue enhances zone edge definition.
- 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.)
BacterLab ensures the quality of each product batch by testing with ATCC reference strains.
| Reference strains | Incubation Conditions | Expected results |
| C. albicans ATCC 10231 | 24 – 48 hours, 35 – 37oC | Good growth |
| S. cerevisiae ATCC 20827 | Good growth |
For 1 liter of medium
| Beef infusion from 300 | 2,0 g |
| Casein Acid Hydrolysate | 1,0 g |
| Starch | 1,5 g |
| Glucose | 10,0 g |
| Methylene Blue | 0,0005 g |
| Agar | 17,0 g |
pH of the ready-to-use medium at 250C: 7,3 ± 0,2
*Note: This ingredient is for reference only our company will formulate the ingredient according to requests or tender specifications.
| Product Name | Code | Packaging |
| BacterPlate™ Mueller Hinton Agar, Glucose-Methylene Blue | 05033 | 10 plates/ box or as per customer request. |
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