[an error occurred while processing this directive]ANAEROBE IDENTIFICATION AGAR BASE
A medium for testing the fermentation capabilities of non-sporing anaerobes.
Code: CB1/1566
Typical formula g/l
Beef extract 4.0
Peptone mixture 16.0 Sodium chloride 5.0 Agar 15.0 pH: 7.2 ± 0.2
Directions
Weigh 40 g of powder, disperse in 1 litre of deionised water. Allow to stand for 10 minutes, swirl to mix then sterilise by autoclaving at 121șC for 15 minutes. Cool to 48șC and aseptically add 50-70mls of sterile defibrinated horse blood. Mix well and pour. Before use, flood the surface with 1 ml of a sterile solution of the substrate under test.
Q.C. organisms:
B. fragilis
Storage: Plates up to 7 days at 2-8șC in the dark. Capped Container up to 3 months at 15-20șC in the dark.
Inoculation: Heavily inoculate a small area of the plate with a loopful of a fresh culture of the test organism. Up to 4 organisms per plate can be tested.
Incubation: 37șC anaerobically for 24-48 hours.
Recognition of fermentation: Remove a small plug of agar from below the growth. Cover the plug with bromothymol blue indicator (0.04%). Colour changes due to production of acidity will develop in a few seconds and should be viewed against a white background. Comparison with controls is useful; a plug taken from an area well away from any growth can be used as a negative control.
References
Phillips K.D. 1976. A Simple and sensitive technique for determining the fermentation reactions of non-sporing anaerobes. J. Appl. Bact. 41: 325-328.