Gut model growth medium Solid components (g/L): Peptone water (2.0), yeast extract (2.0), NaCl (0.1), K2HPO4 (0.04), KH2PO4 (0.04), MgSO4.7H2O (0.01), CaCl2.2H2O (0.01), NaHCO3 (2.0), haemin (0.005), cysteine HCl (0.5), bile salts (0.5), glucose (0.4), arabinogalactan (1.0), pectin (2.0), starch (3.0) Liquid components: vitamin K1 at 10 mL/L, and Tween 80 at 0.2% Medium is sterilised at 121 °C for 30 min and cooled to 50 °C. Post-autoclaving glucose (0.4 g/L), and resazurin anaerobic indicator (0.005 g/L) are added. | |||
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Target populations | Agar | Supplements | Incubation (temp, environment) |
Total anaerobes and total Clostridium spp. | Fastidious anaerobe agar | 5% horse blood | 37 °C, anerobic |
Bifidobacterium spp. | 42.5 g/L Columbia agar, and 5 g/L agar technical | 0.5 g/L cysteine HCl, 5 g/L glucose | 37 °C, anerobic |
Bacteroides spp. | Bacteroides bile aesculin agar | 5 mg/L haemin, 10 μL/L vitamin K, 7.5 mg/L vancomycin, 1 mg/L penicillin, 75 mg/L kanamycin and 10 mg/L colistin | 37 °C, anerobic |
Lactobacillus spp. | 52.2 g/L MRS broth and 20 g/L agar technical | 0.5 g/L cysteine hydrocloride, 20 mg/L vancomycin | 37 °C, anerobic |
Total facultative anaerobes | Nutrient agar | N/A | 37 °C, aerobic |
Lactose fermenting Enterobacteriaceae | MaConkey’s agar | N/A | 37 °C, aerobic |
Enterococcus spp. | Kanamycin aesculin azide agar | 10 mg/L nalidixic acid, 10 mg/L aztreonam, and 20 mg/L kanamycin | 37 °C, aerobic |
Total spores (following alcohol shock for 1 h) | Fastidious anaerobe agar | 5% horse blood | 37 °C, anerobic |
C. difficile total viable cells | Braziers CCEY agar | D-cycloserine (250 mg/L) cefoxitin (8 mg/L), 5 mg/L lysozyme, and 20 mL/L lysed horse blood | 37 °C, anerobic |
C. difficile spores | Braziers CCEY agar | 5 mg/L lysozyme, and 2% lysed horse blood | 37 °C, anerobic |