|
M28a
Campylobacter enrichment broth
(Bolton formula)
(Oxoid AM-7526 (manufactured by Med-Ox Chemicals Ltd.) or Malthus Diagnostics Lab)
Meat Peptone | 10 g |
Lactalbumin Hydrolysate | 5 g |
Yeast Extract | 5 g |
NaCl | 5 g |
Haemin | 0.01 g |
Sodium Pyruvate | 0.5 g |
Alpha-Ketoglutamic Acid | 1 g |
Sodium Metabisulphite | 0.5 g |
Sodium Carbonate | 0.6 g |
Total Dry Incredients | 27.61 g |
Distilled Water | 1000 ml |
Final pH, 7.4 ± 0.2.
THOROUGHLY MIX 27.61 g in 1 liter water. Let soak 10 min. Swirl again and autoclave 15 min at 121°C (in screw-capped bottles if possible). Before use, add 2 rehydrated vials of Campylobacter enrichment broth (Bolton formula) supplement (Oxoid NDX131 or Malthus Diagnostics X-131). If supplement is not available add 4 ml each of antibiotic concentrates (formulas below, solutions made separately).
Store powdered media in a tightly fastened container in a cool, dry area to reduce oxygen infusion and peroxide formation, which can inhibit recovery of microaerophiles. Use prepared broth within 1 month of preparation (preferably less than 2 weeks).
Item | Storage conditions |
Shelf life |
---|---|---|
Complete medium | cool, dry | 2 weeks - 1 month |
Sodium cefoperazone | 4°C | 5 days |
-20°C | 14 days | -70°C | 5 months |
Trimethoprim lactate | 4°C | 1 year |
vancomycin | 4°C | 2 months |
Cycloheximide* | 4°C | 1 year |
*Note: Substitute solubilized amphotericin B (Sigma Cat. No. A9528) for cycloheximide if cycloheximide not available. See instructions below. |
Trimethoprim
Trimethoprim lactate (Sigma Cat. No. T0667). Dissolve 0.66 g in 100 ml distilled water, and filter. May be stored 1 year at 4°C. Add 4 ml/liter to yield final concentration of 20 mg/liter Trimethoprim.
or
Trimethoprim (Sigma Cat. No. T7883)[a lower cost alternative]. To prepare Trimethoprim (TMP)-HCl: add 0.5 g TMP to 30ml 0.05N HCl at 50°C (stir until dissolved using hot plate with magnetic stirrer). Adjust volume to 100ml with distilled water. Add 4 ml/liter to yield final concentration of 20 mg/liter Trimethoprim.
Vancomycin (Sigma). Dissolve 0.5 g in 100 ml distilled water and
filter. Store up to 2 months at 4°C. Because of short shelf life, prepare smaller
amounts. Add 4 ml/liter for final concentration of 20 mg/liter
Cycloheximide Dissolve 1.25 g in 20-30 ml ethanol in a 100 ml volumetric flask and bring to line with water. Filter-sterilize. Store at 4°C up to 1 year. Add 4 ml for final a concentration of 50 mg/liter. Use amphotericin B as described below if cycloheximide is not available.
Note: For information on cycloheximide availability problems, see Advisory for BAM Users on Reported Supply Problems for Cycloheximide
Hypertext updated by rim/kwg/cjm 2005-OCT-21