APPENDIX 1 medium at 37°C for 24 48 hours. Pitting of – Glucose-yeast extract medium the slope gives a positive result for T. pyogenes. Yeast extract 10g NANAT Selective medium for the isolation Glucose 10g of Rhodococcus equi. Agar 15g Base medium Peptone 20g Tap water 1000ml Sodium chloride 5g Yeast extract 5g serum slant Loeffler’s Glucose 5g Sodium hydrogen sulphite 0.2g This medium is used to isolate Sodium thiosulfate 1.2g Corynebacterium diptheriae, but also can be Dipotassium hydrogen 2g used to demonstrate proteolytic activity of T. phosphate 2g pyogenes. Sodium hydrogen carbonate 300g Nutrient broth (horse meat broth) 15g Bovine serum 70ml Agarose 1000ml Infusion dextrose broth 2,5g Water (dried) Whole egg dried 7.5g Dissolve by heating and adjust to pH 7.2, bottle and sterilize at 121°C for 15 min. Distilled water 1000ml Cool to 55°C and add the following 4 Final pH: 7.6 antibiotics to reach the following concentrations Medium preparation Mix, and dispense in glass heat resistant bottle. Arrange so that they are in a slant Novobiocin 25g/ml position and autoclave at 120°C for 15 minutes. Properly prepared slants are smooth actidone [cycloheximide] 40g/ml and greyish-white in colour. potassium tellurite 50g/ml Test procedure nalidixic acid 20g/ml A loopful of the bacterial colony is made in the centre of the slope, taking care not to break Brochothrix selective agar the surface of the medium. Incubate the
APPENDIX 1 Glucose-yeast extract medium Yeast extract 10g Glucose 10g Agar 15g Tap water 1000ml Loeffler’s serum slant This medium is used to isolate Corynebacterium diptheriae, but also can be used to demonstrate proteolytic activity of T. pyogenes. Bovine serum 70ml Infusion dextrose broth (dried) 2,5g Whole egg dried 7.5g Distilled water 1000ml Final pH: 7.6 Medium preparation Mix, and dispense in glass heat resistant bottle. Arrange so that they are in a slant position and autoclave at 120°C for 15 minutes. Properly prepared slants are smooth and greyish-white in colour. Test procedure A loopful of the bacterial colony is made in the centre of the slope, taking care not to break the surface of the medium. Incubate the medium at 37°C for 24 – 48 hours. Pitting of the slope gives a positive result for T. pyogenes. NANAT Selective medium for the isolation of Rhodococcus equi. Base medium Peptone Sodium chloride Yeast extract Glucose Sodium hydrogen sulphite Sodium thiosulfate Dipotassium hydrogen phosphate Sodium hydrogen carbonate Nutrient broth (horse meat broth) Agarose Water 20g 5g 5g 5g 0.2g 1.2g 2g 2g 300g 15g 1000ml Dissolve by heating and adjust to pH 7.2, bottle and sterilize at 121°C for 15 min. Cool to 55°C and add the following 4 antibiotics to reach the following concentrations Novobiocin actidone [cycloheximide] potassium tellurite nalidixic acid 25g/ml 40g/ml 50g/ml 20g/ml Brochothrix selective agar
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