M-CP Agar Base 1035 cally add 80.0mL of sterile egg yolk emulsion, 50%. Mix thoroughly. Pour into sterile Petri dishes in 20.0mL volumes. Use: For the cultivation of Clostridium botulinum. McClung-Toabe Egg Yolk Agar, CDC Modified (CDC Modified McClung-Toabe Egg Yolk Agar) Composition per liter: Pancreatic digest of casein 40.0g Agar 25.0g NaHPO 4 5.0g Yeast extract 5.0g D-Glucose 2.0g NaCl 2.0g Egg yolk emulsion 100.0mL MgSO 4 (5% solution) 0.2mL pH 7.4 ± 0.2 at 25°C Egg Yolk Emulsion: Composition : Chicken egg yolks 11 Whole chicken egg 1 Preparation of Egg Yolk Emulsion: Soak eggs with 1:100 dilu- tion of saturated mercuric chloride solution for 1 min. Crack eggs. Sep- arate yolks from whites for 11 eggs. Mix egg yolks with 1 chicken egg. Preparation of Medium: Add components, except egg yolk emul- sion, to distilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Gently heat while stirring and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 60°C. Aseptically add 100.0mL of sterile egg yolk emulsion. Mix thoroughly. Pour into ster- ile Petri dishes in 20.0mL volumes. Use: For the isolation, cultivation, and differentiation of anaerobic bacteria from foods. Bacteria that produce lecithinase appear as colo- nies surrounded by an insoluble opaque precipitate. Bacteria that pro- duce lipase activity appear as colonies with a sheen or “pearly” surface. Bacteria that possess proteolytic activity appear as colonies surrounded by a clear zone. McClung-Toabe Egg Yolk Agar, CDC Modified (CDC Modified McClung-Toabe Egg Yolk Agar) Composition per liter: Pancreatic digest of casein 40.0g Agar 25.0g Na 2 HPO 4 5.0g Yeast extract 5.0g Glucose 2.0g NaCl 2.0g KH 2 PO 4 1.0g Egg yolk emulsion, 50% 100.0mL MgSO 4 ·7H 2 O (5% solution) 0.2mL pH 7.3 ± 0.2 at 25°C Egg Yolk Emulsion, 50%: Composition per 100.0mL: Chicken egg yolks 11 Whole chicken egg 1 NaCl (0.9% solution) 50.0mL Preparation of Egg Yolk Emulsion, 50%: Soak eggs with 1:100 dilution of saturated mercuric chloride solution for 1 min. Crack eggs and separate yolks from whites. Mix egg yolks with 1 chicken egg. Beat to form emulsion. Measure 50.0mL of egg yolk emulsion and add to 50.0mL of 0.9% NaCl solution. Mix thoroughly. Filter sterilize. Warm to 45°–50°C. Preparation of Medium: Add components—except egg yolk emulsion, 50%—to distilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Gently heat while stirring and bring to boil- ing. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Aseptically add 100.0mL of sterile egg yolk emulsion, 50%. Mix thor- oughly. Pour into sterile Petri dishes in 15.0mL volumes. Use: For the cultivation of a wide variety of anaerobic bacteria. For the differentiation of anaerobic bacteria based on lecithinase production, lipase production, and proteolytic ability. Bacteria that produce lecithi- nase appear as colonies surrounded by a zone of insoluble precipitate. Bacteria that produce lipase appear as colonies with a pearly iridescent sheen. Bacteria that produce proteolytic activity appear as colonies sur- rounded by a clear zone. McClung Toabe HiVeg Agar Base wtih Egg Yolk Composition per liter: Plant peptone No. 3 40.0g Agar 25.0g Na 2 HPO 4 5.0g Glucose 2.0g NaCl 2.0g KH 2 PO 4 1.0g MgSO 4 0.1g Egg yolk emulsion, 50% 100.0mL pH 7.3 ± 0.2 at 25°C Source: This medium, without egg yolk emulsion, is available as a premixed powder from HiMedia. Egg Yolk Emulsion, 50%: Composition per 100.0mL: Chicken egg yolks 11 Whole chicken egg 1 NaCl (0.9% solution) 50.0mL Preparation of Egg Yolk Emulsion, 50%: Soak eggs with 1:100 dilution of saturated mercuric chloride solution for 1 min. Crack eggs and separate yolks from whites. Mix egg yolks with 1 chicken egg. Beat to form emulsion. Measure 50.0mL of egg yolk emulsion and add to 50.0mL of 0.9% NaCl solution. Mix thoroughly. Filter sterilize. Warm to 45°–50°C. Preparation of Medium: Add components, except egg yolk emul- sion, 50%, to distilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Gently heat while stirring and bring to boiling. Auto- clave for 20 min at 15 psi pressure–121°C. Cool to 50°–55°C. Asepti- cally add 100.0mL of sterile egg yolk emulsion, 50%. Mix thoroughly. Pour into sterile Petri dishes in 15.0mL volumes. Use: For the isolation and cultivation of Clostridium perfringens in foods. M-CP Agar Base Composition per liter: Tryptose 30.0g Yeast extract 20.0g Agar 15.0g Sucrose 5.0g L-Cysteine·HCl·H 2 O 1.0g MgSO 4 ·7H 2 O 0.1g FeCl 3 ·6H 2 O 0.09g Indoxyl β- D-glucoside 0.06g © 2010 by Taylor and Francis Group, LLC 1036 M-CP HiVeg Agar Base with Phenolphthalein Diphosphate Bromcresol Purple 0.04g Selective supplement solution B 20.0mL Selective supplement solution A 10.0mL pH 7.6 ± 0.2 at 25°C Source: This medium is available from HiMedia. Selective Supplement Solution A: Composition per 10.0mL: D-Cycloserine 400.0mg Polymyxin B sulfate 25.0mg Preparation of Selective Supplement Solution A: Add compo- nents to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Selective Supplement Solution B: Composition per 20.0mL: Phenolphthalein diphosphate 0.1g Preparation of Selective Supplement Solution B: Add phenol- phthalein diphosphate to distilled/deionized water and bring volume to 20.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except selective sup- plement solutions A and B, to distilled/deionized water and bring vol- ume to 970.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Aseptically add selective supplement solutions A and B. Mix thoroughly. Pour into Petri dishes or aseptically distribute into sterile tubes. Use: Recommended by the Directive of the Council of the European Union 98/83/EC for the isolation and enumeration of Clostridium per- fringens from water samples using the membrane filtration technique. M-CP HiVeg Agar Base with Phenolphthalein Diphosphate Composition per liter: Plant hydrolysate No. 1 30.0g Agar 15.0g L-Cysteine·HCl 1.0g MgSO 4 ·7H 2 O 0.1g FeCl 3 ·6H 2 O 0.09 Indoxyl β-D-glucoside 0.06 Bromcresol Purple 0.04 Sucrose 5.0g Yeast extract 20.0g Phenolphthalein diphosphate solution 10.0mL pH 7.2 ± 0.2 at 25°C Source: This medium, without phenolphthalein diphosphate, is avail- able as a premixed powder from HiMedia. Phenolphthalein Diphosphate Solution: Composition per 10.0mL: Phenolphthalein diphosphate 2.0g Preparation of Phenolphthalein Diphosphate Solution: Add phenolphthalein diphosphate to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except phenolphtha- lein diphosphate solution, to distilled/deionized water and bring vol- ume to 990.0mL. Mix thoroughly. Gently heat while stirring until boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45- 50°C. Aseptially add 10.0mL of sterile phenolphthalein diphosphate solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and identification of Providencia stuartii. m-CP Medium Composition per liter: Tryptose 30.0g Yeast extract 20.0g Agar 15.0g Sucrose 5.0g L-Cysteine·HCl·H 2 O 1.0g MgO 4 ·7H 2 O 0.1g Bromcresol Purple 0.04g Phenolphthalein biphosphate tetrazolium salt solution 10.0mL Selective supplement solution 4.0mL Indoxyl-β-D-glucoside solution 4.0mL Ferric chloride solution 1.0mL pH 7.6 ± 0.2 at 25°C Selective Supplement Solution: Composition per 4.0mL: D-Cycloserine 0.4g Polymyxin B sulfate 25.0mg Preparation of Selective Supplement Solution: Add compo- nents to 4.0mL of distilled/deionized water. Mix thoroughly. Filter ster- ilize. Phenolphthalein Biphosphate Tetrazolium Salt Solution: Composition per 10.0mL: Phenolphthalein biphosphate tetrazolium salt 25.0mg Preparation of Phenolphthalein Biphosphate Tetrazolium Salt Solution: Add phenolphthalein biphosphate tetrazolium salt to 10.0mL of distilled/deionized water. Mix thoroughly. Filter sterilize. Indoxyl-β-D-glucoside Solution: Composition per 10.0mL: Indoxyl-β- D-glucoside 0.45g Preparation of Indoxyl-β-D-glucoside Solution: Add indoxyl- β- D-glucoside to 10.0mL of distilled/deionized water. Mix thoroughly. Filter sterilize. Ferric Chloride Solution: Composition per 4.0mL: FeCl 3 ·6H 2 O 30.0mg Preparation of Ferric Chloride Solution: Add FeCl 3 ·6H 2 O to 4.0mL of distilled/deionized water. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except selective sup- plement solution, phenolphthalein biphosphate tetrazolium salt solu- tion, indoxyl-β- D-glucoside solution, and ferric chloride solution, to distilled/deionized water and bring volume to 981.0mL. Mix thorough- ly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add 4.0mL of selective supplement solution. Mix thor- oughly. Aseptically add 10.0mL phenolphthalein biphosphate tetrazo- lium salt solution, 4.0mL indoxyl-β- D-glucoside solution, and 1.0mL ferric chloride solution. Mix thoroughly. Pour into sterile Petri dishes or aseptically distribute into tubes. Use: A selective, chromogenic medium for the rapid identification and enumeration of Clostridium perfringens in water samples, including water used in food and beverage production. © 2010 by Taylor and Francis Group, LLC MD1- Medium 1037 MCP Medium (Modified MacConkey Medium) (MacConkey Phosphatase Medium) Composition per liter: Pancreatic digest of gelatin 17.0g Agar 13.5g Lactose 10.0g NaCl 5.0g Bile salts 1.5g Pancreatic digest of casein 1.5g Peptic digest of animal tissue 1.5g Na 2 HPO 4 0.6g Glucose 0.2g Methyl Blue 0.07g Neutral Red 0.03g Crystal Violet 1.0mg Phenolphthalein diphosphate solution 10.0mL pH 7.2 ± 0.2 at 25°C Source: This medium, without phenolphthalein diphosphate, is avail- able as a premixed powder from HiMedia. Phenolphthalein Diphosphate Solution: Composition per 10.0mL: Phenolphthalein diphosphate 2.0g Preparation of Phenolphthalein Diphosphate Solution: Add phenolphthalein diphosphate to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except phenolphtha- lein diphosphate solution, to distilled/deionized water and bring vol- ume to 990.0mL. Mix thoroughly. Gently heat while stirring until boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45- 50°C. Aseptially add 10.0mL of sterile phenolphthalein diphosphate solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and identification of Providencia stuartii. MD Medium Composition per liter: Agar 20.0g L-Malic acid 20.0g Pancreatic digest of casein 10.0g D-Glucose 5.0g Casamino acids 3.0g Pancreatic digest of soybean meal 1.5g Tween™ 80 1.0g Yeast extract 1.0g Bromcresol Green solution 20.0mL pH 7.0 ± 0.2 at 25°C Bromcresol Green Solution: Composition per 30.0mL: Bromcresol Green 0.1g NaOH (0.01N solution) 30.0mL Preparation of Bromcresol Green Solution: Add Bromcresol Green to 30.0mL of NaOH solution. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Adjust pH to 7.0 with 10N KOH. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the isolation and cultivation of Salmonella species from foods. MD 1 Medium Composition per liter: Pancreatic digest of casein 3.0g MgSO 4 ·7H 2 O 2.0g CaCl 2 0.5g Trace elements solution 1.0mL Vitamin B 12 solution 1.0mL Trace Elements Solution: Composition per liter: EDTA 8.0g MnCl 2 ·4H 2 O 0.1g CoCl 2 0.02g KBr 0.02g ZnCl 2 0.02g CuSO 4 0.01g H 3 BO 3 0.01g NaMoO 4 ·2H 2 O 0.01g BaCl 2 5.0mg LiCl 5.0mg SnCl 2 ·2H 2 O 5.0mg Preparation of Trace Elements Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Vitamin B 12 Solution: Composition per 10.0mL: Vitamin B 12 5.0mg Preparation of Vitamin B 12 Solution: Add vitamin B 12 to dis- tilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of myxobacteria. MD1- Medium (DSMZ Medium 1118) Composition per liter: Casitone 3.0g MgSO 4 ·7H 2 O 2.0g CaCl 2 ·2H 2 O 0.7g Vitamin solution 10.0mL Trace elements solution SL-4 1.0mL pH 7.1 ± 0.2 at 25°C Trace Elements Solution SL-4: Composition per liter: EDTA 0.5g FeSO 4 ·7H 2 O 0.2g Trace elements solution SL-6 100.0mL Trace Elements Solution SL-6: Composition per liter: MnCl 2 ·4H 2 O 0.5g H 3 BO 3 0.3g CoCl 2 ·6H 2 O 0.2g ZnSO 4 ·7H 2 O 0.1g Na 2 MoO 4 ·2H 2 O 0.03g © 2010 by Taylor and Francis Group, LLC 1038 MDPA with Calcium Carbonate NiCl 2 ·6H 2 O 0.02g CuCl 2 ·2H 2 O 0.01g Preparation of Trace Elements Solution SL-6: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Trace Elements Solution SL-4: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize. Vitamin Solution: Composition per 10.0ml: Vitamin B 12 0.5mg Preparation of Vitamin Solution: Add components to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except vitamin solu- tion, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Adjust pH to 7.1. Distribute into tubes or flasks. Gently heat while stirring and bring to boiling. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically add 10.0mL sterile vita- min solution. Mix thorougly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation of Myxococcus xanthus. MDPA See: Malt Dextrose Peptone Agar MDPA with Calcium Carbonate Composition per liter: Agar 25.0g Malt extract 20.0g Glucose 20.0g CaCO 3 5.0g Peptone 1.0g Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation of Dekkera species. MDPYA4 See: Malt 4% Dextrose Peptone Yeast Agar MDYA4 See: Malt 4% Dextrose Yeast Agar m-E Agar See: E Agar Me15% MH Agar (DSMZ Medium 582) Composition per liter: NaCl 121.5g Agar 20.0g MgSO 4 14.4g MgCl 2 10.5g Yeast extract 10.0g Proteose peptone no. 3 5.0g KCl 3.0g Glucose 1.0g CaCl 2 0.54g NaBr 0.039g NaHCO 3 solution 10.0mL pH 7.5 ± 0.2 at 25°C NaHCO 3 Solution: Composition per 100.0mL: NaHCO 3 0.9g Preparation of NaHCO 3 Solution: Add NaHCO 3 to distilled/de- ionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure– 121°C. Cool to 50°C. Preparation of Medium: Add components, except NaHCO 3 solu- tion, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Aseptically add 10.0mL NaHCO 3 solution. Mix thoroughly. Pour into Petri dishes or aseptically distribute into sterile tubes. Use: For the cultivation of Bacillus halophilus. Me15% MH Medium (DSMZ Medium 582) Composition per liter: NaCl 121.5g MgSO 4 14.4g MgCl 2 10.5g Yeast extract 10.0g Proteose peptone no. 3 5.0g KCl 3.0g Glucose 1.0g CaCl 2 0.54g NaBr 0.039g NaHCO 3 solution 10.0mL pH 7.5 ± 0.2 at 25°C NaHCO 3 Solution: Composition per 100.0mL: NaHCO 3 0.9g Preparation of NaHCO 3 Solution: Add NaHCO 3 to distilled/de- ionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure– 121°C. Cool to 25°C. Preparation of Medium: Add components, except NaHCO 3 solu- tion, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Aseptically add 10.0mL NaHCO 3 solution. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation of Bacillus halophilus. MEA See: Malt Extract Agar Meat Extract with Peptone (Pepted Meat Broth) Composition per liter: NaCl 15.0g Peptic digest of animal tissue 10.0g Meat extract 3.0g pH 7.2 ± 0.2 at 25°C © 2010 by Taylor and Francis Group, LLC Medium A for Producing Lysates 1039 Source: This medium is available from HiMedia. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Alcaligenes species. Meat Extract with Peptone and 1.5% Salt Composition per liter: NaCl 15.0g Peptone 10.0g Meat extract 3.0g Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation and maintenance of Alcaligenes species. Meat Infusion Agar, HiVeg (Standard Infusion Agar, HiVeg) Composition per liter: Agar 25.0g Plant infusion 10.0g Plant peptone 10.0g NaCl 5.0g pH 7.5 ± 0.2 at 25°C Source: This medium is available as a premixed powder from Hi- Media. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the mass cultivation of organisms for vaccine or toxin pro- duction. M-EC Test Agar Composition per liter: Agar 15.0g Lactose 10.0g NaCl 7.5g Proteose peptone 5.0g Dipotassium phosphate 3.3g Yeast extract 3.0g KH 2 PO 4 1.0g Sodium lauryl sulphate 0.2g Sodium deoxycholate 0.1g Bromcresol Purple 0.08g Bromphenol Red 0.08g pH 7.3 ± 0.2 at 25°C Source: This medium is available from HiMedia. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Do not autoclave. Pour into sterile Petri dishes or leave in tubes. Use: For the detection of Escherichia coli in water samples using the membrane filter technique. MED IIa Composition per liter: Tris buffer stock solution 10.0mL CaCl 2 (5.0% solution) 10.0mL MgSO 4 ·7H 2 O (3.33% solution) 1.0mL pH 7.2 ± 0.2 at 25°C Tris Buffer Stock Solution: Composition per 500.0mL: Tris(hydroxymethyl)aminomethane·HCl 35.01g Tris(hydroxymethyl)aminomethane 3.35g Preparation of Tris Buffer Stock Solution: Add components to distilled/deionized water and bring volume to 500.0mL. Mix thorough- ly. Adjust pH to 7.2. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 20 min at 15 psi pressure–121°C. Use: For the cultivation and maintenance of Vampirovibrio chlorellavo- rus. Medium A Composition per liter: D-Glucose 20.0g Agar 20.0g Yeast extract 10.0g Biotin 1.0mg Calcium pantothenate 1.0mg pH 7.3 ± 0.2 at 25°C Preparation of Medium: Add components, except biotin and cal- cium pantothenate, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Add biotin and calcium pantothenate to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Aseptically add the sterile bi- otin and calcium pantothenate solution to the cooled sterile basal me- dium. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of Zymomonas mobilis. Medium A for Producing Lysates Composition per liter: Nutrient broth 8.0g KCl 1.0g MgSO 4 ·7H 2 O 0.25g MnCl 2 1.25mg FeSO 4 (1.0mM solution) 1.0mL Ca(NO 3 ) 2 (1.0M solution) 1.0mL pH 7.0–7.2 at 25°C Preparation of Medium: Add components, except FeSO 4 and Ca(NO 3 ) 2 , to distilled/deionized water and bring volume to 998.0mL. Mix thoroughly. Adjust pH to 7.0–7.2. Autoclave for 30 min at 15 psi pressure–115°C. Cool to 45°–50°C. Prepare 1.0mM FeSO 4 solution and 1.0M Ca(NO 3 ) 2 solution separately. Filter sterilize both solutions. Aseptically add the sterile FeSO 4 solution and sterile Ca(NO 3 ) 2 solu- tion to the cooled sterile basal medium. Mix thoroughly. Distribute into sterile tubes or flasks. Use: For the cultivation of microorganisms to be lysed. © 2010 by Taylor and Francis Group, LLC 1040 Medium 2A Medium 2A Composition per liter: Arginine 10.0g NaCl 5.0g Agar 4.0g Peptone 1.0g K 2 HPO 4 ·3H 2 O 0.3g Phenol Red 0.01g pH 7.2–7.4 at 25°C Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes. Autoclave for 15 min at 15 psi pres- sure–121°C. Use: For the cultivation and differentiation of Pseudomonas species based on their production of arginine dihydrolase activity. Medium AS4 Composition per liter: Sucrose 80.0g PPLO broth without Crystal Violet 500.0mL Horse serum 200.0mL Phenol Red (0.5% solution) 5.0mL pH 7.2 ± 0.2 at 25°C PPLO Broth without Crystal Violet: Composition per 500.0mL: Beef heart, solids from infusion 11.53g Peptone 2.33g NaCl 1.15g Source: PPLO broth without Crystal Violet is available as a premixed powder from BD Diagnostic Systems. Preparation of PPLO Broth without Crystal Violet: Add components to distilled/deionized water and bring volume to 500.0mL. Mix thoroughly. Beef heart for infusion may be substituted; 100.0g of beef heart for infusion is equivalent to 500.0g of fresh heart tissue. Preparation of Medium: Add components, except horse serum, to distilled/deionized water and bring volume to 800.0mL. Mix thorough- ly. Adjust pH to 7.2. Autoclave for 10 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add 200.0mL of noninactivated, sterile horse serum. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation and maintenance of Spiroplasma melliferum. Medium for Acetivibrio cellulolyticus See: BC Medium Medium for Aciduric, Thermophilic Bacillus Strains Composition per liter: Solution A 500.0mL Solution B 500.0mL pH 4.3 ± 0.2 at 25°C Solution A: Composition per 500.0mL: KH 2 PO 4 3.0g Glucose 1.0g Starch 1.0g Tryptone 1.0g Yeast extract 1.0g MgSO 4 ·7H 2 O 0.5g CaCl 2 ·2H 2 O 0.25g (NH 4 ) 2 SO 4 0.2g Preparation of Solution A: Add components to distilled/deionized water and bring volume to 500.0mL. Mix thoroughly. Adjust pH to 4.3. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Solution B: Composition per 500.0mL: Agar 20.0g Preparation of Solution B: Add agar to distilled/deionized water and bring volume to 500.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Preparation of Medium: Aseptically combine 500.0mL of solu- tion A and 500.0mL of solution B. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation of aciduric, thermophilic Bacillus strains. Medium 2508-85-1 with Amino Acids Composition per liter: Agar 20.0g Nutrient broth 8.0g D-Glucose 5.0g Polypeptone™ 5.0g Yeast extract 5.0g L-Lysine 0.1g L-Methionine 0.05g Diaminopimelic acid 0.05g Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 30 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Escherichia coli. Medium for Ammonia Oxidizers Composition per liter: MgSO 4 ·7H 2 O 0.2g (NH 4 ) 2 SO 4 0.13g K 2 HPO 4 0.09g CaCl 2 ·2H 2 O 0.02g Chelated iron 1.0mg MnCl 2 ·4H 2 O 0.2mg Na 2 MoO 4 ·2H 2 O 0.1mg ZnSO 4 ·7H 2 O 0.1mg CuSO 4 ·5H 2 O 0.02mg CoCl 2 ·6H 2 O 2.0μg Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the isolation, cultivation, and enrichment of ammonia-oxi- dizing bacteria from soil. Medium for Ammonia Oxidizers Composition per liter: (NH 4 ) 2 SO 4 2.0g MgSO 4 ·7H 2 O 0.2g CaCl 2 ·2H 2 O 0.02g K 2 HPO 4 0.02g Chelated iron 1.0mg © 2010 by Taylor and Francis Group, LLC Medium for Ammonia-Oxidizing Bacteria 1041 MnCl 2 ·4H 2 O 0.2mg Na 2 MoO 4 ·2H 2 O 0.1mg ZnSO 4 ·7H 2 O 0.1mg CuSO 4 ·5H 2 O 0.02mg CoCl 2 ·6H 2 O 2.0μg Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the isolation, cultivation, and enrichment of ammonia-oxi- dizing bacteria from soil. Medium for Ammonia Oxidizers Composition per liter: K 2 HPO 4 0.5g (NH 4 ) 2 SO 4 0.5g Phenol Red 0.5g MgSO 4 ·7H 2 O 0.05g CaCl 2 ·2H 2 O 0.02g NaCl 0.02g Na 2 MoO 4 ·2H 2 O 2.4μg Metals “44” 1.0mL Metals “44”: Composition per 100.0mL: ZnSO 4 ·7H 2 O 1.1g FeSO 4 ·7H 2 O 0.5g EDTA 0.25g MnSO 4 ·7H 2 O 0.154g CuSO 4 ·5H 2 O 0.04g Co(NO 3 ) 2 ·6H 2 O 0.025g Na 2 B 4 O 7 ·10H 2 O 0.018g Preparation of Metals “44”: Add a few drops of H 2 SO 4 to dis- tilled/deionized water to inhibit precipitate formation. Add compo- nents to acidified distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the isolation, cultivation, and enrichment of ammonia-oxi- dizing bacteria from soil. Medium for Ammonia Oxidizers Composition per liter: (NH 4 ) 2 SO 4 0.5g KH 2 PO 4 0.2g CaCl 2 ·2H 2 O 0.04g MgSO 4 ·7H 2 O 0.04g Ferric citrate 0.5mg Phenol Red 0.5mg Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the isolation, cultivation, and enrichment of ammonia-oxi- dizing bacteria from soil. Medium for Ammonia Oxidizers, Brackish Composition per liter: CaCO 3 5.0g NH 4 Cl 0.5g K 2 HPO 4 0.05g Seawater 400.0mL Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the isolation, cultivation, and enrichment of ammonia-oxidiz- ing bacteria from brackish specimens. Medium for Ammonia Oxidizers, Marine Composition per liter: (NH 4 ) 2 SO 4 1.32g MgSO 4 ·7H 2 O 0.2g Chelated iron 0.13g K 2 HPO 4 0.11g CaCl 2 ·2H 2 O 0.02g ZnSO 4 ·7H 2 O 0.1mg CuSO 4 ·5H 2 O 0.02mg CoCl 2 ·6H 2 O 2.0μg MnCl 2 ·4H 2 O 2.0μg Na 2 MoO 4 ·2H 2 O 1.0μg Seawater 1.0L Preparation of Medium: Combine components. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure– 121°C. Use: For the isolation, cultivation, and enrichment of marine ammo- nia-oxidizing bacteria. Medium for Ammonia-Oxidizing Bacteria Composition per liter: (NH 4 ) 2 SO 4 235.0mg KH 2 PO 4 200.0mg CaCl 2 ·2H 2 O 40.0mg MgSO 4 ·7H 2 O 40.0mg Iron-EDTA-Phenol Red solution 1.0mL Na 2 CO 3 solution variable Na 2 CO 3 Solution: Composition per 100.0mL: Na 2 CO 3 5.0g Preparation of Na 2 CO 3 Solution: Add Na 2 CO 3 to distilled/de- ionized water and bring volume to 100.0mL. Mix thoroughly. Auto- clave for 15 min at 15 psi pressure–121°C. Iron-EDTA-Phenol Red Solution: Composition per 100.0mL: FeSO 4 ·7H 2 O 50.0mg Sodium EDTA 50.0mg Phenol Red 50.0mg Preparation of Iron-EDTA-Phenol Red Solution: Add compo- nents to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Preparation of Medium: Add components, except Na 2 CO 3 solu- tion, to distilled/deionized water and bring volume to 1.0L. Mix thor- oughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Add enough sterile Na 2 CO 3 solution to turn the medi- um pale pink. During incubation and growth of bacteria, add additional sterile Na 2 CO 3 solution to restore the pale pink color. Growth is com- plete when no further color change is observed. © 2010 by Taylor and Francis Group, LLC 1042 Medium B for Sulfate Reducers Use: For the cultivation of Nitrosolobus multiformis and Nitrosomo- nas europaea. Medium B for Sulfate Reducers (Postgate’s Medium B for Sulfate Reducers) Composition per liter: Sodium lactate 3.5g MgSO 4 ·7H 2 O 2.0g NH 4 Cl 1.0g CaSO 4 1.0g Yeast extract 1.0g KH 2 PO 4 0.5g FeSO 4 ·7H 2 O 0.5g Ascorbic acid 0.1g Thioglycollic acid 0.1g pH 7.0–7.5 at 25°C Preparation of Medium: Add components, except ascorbic acid and thioglycollic acid, to tap water and bring volume to 1.0L. For ma- rine bacteria, NaCl may be added or seawater used in place of tap wa- ter. Mix thoroughly. Adjust pH to 7.0–7.5. Thioglycolate and ascorbate should be added immediately prior to sterilization. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the isolation, cultivation, and maintenance of Desulfovibrio species and Desulfotomaculum species. This medium turns black as a result of H 2 S production due to bacterial growth. Medium for Bacillus schlegelii Composition per liter: Agar 15.0g Na 2 HPO 4 ·2H 2 O 2.9g KH 2 PO 4 2.3g NH 4 Cl 1.0g MgSO 4 ·7H 2 O 0.5g NaHCO 3 0.5g CaCl 2 ·2H 2 O 0.01g MnSO 4 ·H 2 O 10.0mg Ferric ammonium citrate solution 20.0mL Trace elements solution SL-6 5.0mL pH 6.8 ± 0.2 at 25°C Ferric Ammonium Citrate Solution: Composition per 20.0mL: Ferric ammonium citrate 0.05g Preparation of Ferric Ammonium Citrate Solution: Add fer- ric ammonium citrate to distilled/deionized water and bring volume to 20.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure– 121°C. Trace Elements Solution SL-6: Composition per liter: MnCl 2 ·4H 2 O 0.5g H 3 BO 3 0.3g CoCl 2 ·6H 2 O 0.2g ZnSO 4 ·7H 2 O 0.1g Na 2 MoO 4 ·2H 2 O 0.03g NiCl 2 ·6H 2 O 0.02g CuCl 2 ·2H 2 O 0.01g Preparation of Trace Elements Solution SL-6: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Preparation of Medium: Add components, except ferric ammoni- um citrate solution, to distilled/deionized water and bring volume to 980.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically add 20.0mL of sterile ferric ammonium citrate solution. Mix thoroughly. Pour into sterile Pe- tri dishes or distribute into sterile tubes. Use: For the chemolithotrophic growth of Bacillus schlegelii. Medium for Bacillus stearothermophilus Composition per 1001.0mL: NH 4 Cl 1.0g K 2 HPO 4 0.5g Yeast extract 0.2g Casamino acids 0.1g MgSO 4 ·7H 2 O 0.02g Phenol solution 100.0mL Trace elements solution SL-4 1.0mL pH 7.4 ± 0.2 at 25°C Phenol Solution: Composition per 100.0mL: Phenol 0.47g Preparation of Phenol Solution: Add phenol to distilled/deion- ized water and bring volume to 100.0mL. Mix thoroughly. Filter ster- ilize. Trace Elements Solution SL-4: Composition per liter: EDTA 0.5g FeSO 4 ·7H 2 O 0.2g Trace elements solution SL-6 100.0mL Trace Elements Solution SL-6: Composition per liter: MnCl 2 ·4H 2 O 0.5g H 3 BO 3 0.3g CoCl 2 ·6H 2 O 0.2g ZnSO 4 ·7H 2 O 0.1g Na 2 MoO 4 ·2H 2 O 0.03g NiCl 2 ·6H 2 O 0.02g CuCl 2 ·2H 2 O 0.01g Preparation of Trace Elements Solution SL-6: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Trace Elements Solution SL-4: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except phenol solution and trace elements solution SL-4, to distilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically add 100.0mL of sterile phenol solu- tion and 1.0mL of sterile trace elements solution SL-4. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation of Bacillus stearothermophilus. Medium BG11 for Cyanobacteria See: BG11 Agar and BG11 Medium Medium BG11 for Marine Cyanobacteria See: BG11 Marine Agar and BG11 Marine Broth © 2010 by Taylor and Francis Group, LLC Medium for Carbon Monoxide Oxidizers 1043 Medium with Biphenyl (DSMZ Medium 457d) Composition per liter: Na 2 HPO 4 2.44g KH 2 PO 4 1.52g (NH 4 ) 2 SO 4 0.5g MgSO 4 ·7H 2 O 0.2g CaCl 2 ·2H 2 O 0.05g Trace elements solution SL-4 10.0mL Biphenyl solution 25.0mL pH 6.9 ± 0.2 at 25°C Trace Elements Solution SL-4: Composition per liter: EDTA 0.5g FeSO 4 ·7H 2 O 0.2g Trace elements solution SL-6 100.0mL Trace Elements Solution SL-6: Composition per liter: H 3 BO 3 0.3g CoCl 2 ·6H 2 O 0.2g ZnSO 4 ·7H 2 O 0.1g MnCl 2 ·4H 2 O 0.03g Na 2 MoO 4 ·H 2 O 0.03g NiCl 2 ·6H 2 O 0.02g CuCl 2· ·2H 2 O 0.01g Preparation of Trace Elements Solution SL-6: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 3.4. Preparation of Trace Elements Solution SL-4: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Biphenyl Solution: Composition per liter: Biphenyl 10.0g Preparation of Biphenyl Solution: Add biphenyl to 1.0L ethanol. Mix thoroughly. Filter sterilize using a cellulose filter membrane. Preparation of Medium: Add components, except biphenyl solu- tion, to 1.0L distilled/deionized water. Adjust pH to 6.9. Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature. Add an al- iquot of the biphenyl solution to a sterile flask so that the final concen- tration will be 0.25g/L biphenyl, and let the ethanol evaporate. Aseptically add sterile medium to the crystal-layered flask. Use: For the cultivation of biphenyl- utilizing bacteria. Medium C for Sulfate Reducers (Postgate’s Medium C for Sulfate Reducers) Composition per liter: Sodium lactate 6.0g Na 2 SO 4 4.5g NH 4 Cl 1.0g Yeast extract 1.0g KH 2 PO 4 0.5g Sodium citrate·2H 2 O 0.3g CaCl 2 ·6H 2 O 0.06g MgSO 4 ·7H 2 O 0.06g FeSO 4 ·7H 2 O 0.004g pH 7.5 ± 0.2 at 25°C Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. For marine bacteria, NaCl may be added or sea water used in place of distilled/deionized water. Mix thor- oughly. Adjust pH to 7.5. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For detection, culturing, and storage of Desulfovibrio species and many Desulfotomaculum species. This medium should be used when a clear culture medium is desired such as for chemostat culture. This medium may be cloudy after sterilization but usually clears on cooling. It turns black as a result of H 2 S production due to bacterial growth. Medium for Campylobacter DSM 806 (DSMZ Medium 121) Composition per liter: Na-aspartate 10.0g MgSO 4 ·7H 2 O 1.0g Yeast extract 0.2g CaCl 2 ·2H 2 O 28.0mg Resazurin 1.0mg Cysteine phosphate solution 100.0mL Trace elements solution SL-10 1.0mL pH 7.0 ± 0.2 at 25°C Trace Elements Solution SL-10: Composition per liter: FeCl 2 ·4H 2 O 1.5g CoCl 2 ·6H 2 O 190.0mg MnCl 2 ·4H 2 O 100.0mg ZnCl 2 70.0mg Na 2 MoO 4 ·2H 2 O 36.0mg NiCl 2 ·6H 2 O 24.0mg H 3 BO 3 6.0mg CuCl 2 ·2H 2 O 2.0mg HCl (25% solution) 10.0mL Preparation of Trace Elements Solution SL-10: Add FeCl 2 ·4H 2 O to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized water and bring volume to 1.0L. Add remaining components. Mix thor- oughly. Gas under 80% N 2 + 20% CO 2 . Cysteine Phosphate Solution: Composition per 100.0mL: K 2 HPO 4 0.75g NaH 2 PO 4 0.25g Cysteine-HCl·H 2 O 0.25g Preparation of Cysteine Phosphate Solution: Add components to 100.0mL distilled/deionized water. Mix thoroughly. Gas under 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: Add components, except cysteine phos- phate solution, to distilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Adjust pH to 7.0. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Aseptically add 100.0mL of sterile cysteine phosphate solution. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation of Sulfurospirillum sp. Medium for Carbon Monoxide Oxidizers Composition per liter: Agar 12.0g Na 2 HPO 4 ·12H 2 O 4.5g NH 4 Cl 1.5g KH 2 PO 4 0.75g MgSO 4 ·7H 2 O 0.2g © 2010 by Taylor and Francis Group, LLC 1044 Medium for Carbon Monoxide Oxidizers CaCl 2 ·2H 2 O 0.03g Ferric ammonium citrate 0.018g Trace elements solution SL-6 1.0mL pH 7.0 ± 0.2 at 25°C Trace Elements Solution SL-6: Composition per liter: MnCl 2 ·4H 2 O 0.5g H 3 BO 3 0.3g CoCl 2 ·6H 2 O 0.2g ZnSO 4 ·7H 2 O 0.1g Na 2 MoO 4 ·2H 2 O 0.03g NiCl 2 ·6H 2 O 0.02g CuCl 2 ·2H 2 O 0.01g Preparation of Trace Elements Solution SL-6: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or distribute into sterile tubes. After inoculation, in- cubate in an atmosphere of 80% CO +10% O 2 + 10% N 2 . Use: For the chemoautotrophic cultivation and maintenance of Alcal- igenes species, Pseudomonas carboxydohydrogena, and other Pseudo- monas species. Medium for Carbon Monoxide Oxidizers Composition per liter: Agar 12.0g Na 2 HPO 4 ·12H 2 O 4.5g Sodium acetate 3.0g NH 4 Cl 1.5g KH 2 PO 4 0.75g MgSO 4 ·7H 2 O 0.2g CaCl 2 ·2H 2 O 0.03g Ferric ammonium citrate 0.018g Trace elements solution SL-6 1.0mL pH 7.0 ± 0.2 at 25°C Trace Elements Solution SL-6: Composition per liter: MnCl 2 ·4H 2 O 0.5g H 3 BO 3 0.3g CoCl 2 ·6H 2 O 0.2g ZnSO 4 ·7H 2 O 0.1g Na 2 MoO 4 ·2H 2 O 0.03g NiCl 2 ·6H 2 O 0.02g CuCl 2 ·2H 2 O 0.01g Preparation of Trace Elements Solution SL-6: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or distribute into sterile tubes. After inoculation in- cubate in air. Use: For the chemoorganotrophic cultivation and maintenance of Alcaligenes species, Pseudomonas carboxydohydrogena, and other Pseudomonas species. Medium for Chlorate Respirers (DSMZ Medium 908) Composition per liter: Solution A 1.0L Solution B 10.0mL Solution C 10.0mL Vitamin solution 5.0mL Trace elements solution SL-10 1.0mL pH 7.2 ± 0.2 at 25°C Solution A: Composition per liter: NaHCO 3 2.5g Na-acetate 1.36g NaClO 3 1.0g NaH 2 PO 4 0.6g NH 4 Cl 0.25g KCl 0.1g Preparation of Solution A: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature. Solution B: Composition per 10.0mL: MgSO 4 30.0mg CaCl 2 ·2H 2 O 10.0mg Preparation of Solution B: Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature. Solution C: Na 2 MoO 4 25.0mg Na 2 WO 4 ·2H 2 O 25.0mg Preparation of Solution C: Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature. Vitamin Solution: Composition per liter: Vitamin B 12 50.0mg Pantothenic acid 50.0mg Riboflavin 50.0mg Alpha-lipoic acid 50.0mg p-Aminobenzoic acid 50.0mg Thiamine-HCl·2H 2 O 50.0mg Nicotinic acid 25.0mg Nicotinamide 25.0mg Biotin 20.0mg Folic acid 20.0mg Pyridoxamine-HCl 10.0mg Preparation of Vitamin Solution: Add components to distilled/ deionized water and bring volume to 1.0L. Mix thoroughly. Filter ster- ilize. Trace Elements Solution SL-10: Composition per liter: FeCl 2 ·4H 2 O 1.5g H 3 BO 3 300.0mg CoCl 2 ·6H 2 O 190.0mg MnCl 2 ·4H 2 O 100.0mg © 2010 by Taylor and Francis Group, LLC . to form emulsion. Measure 50.0mL of egg yolk emulsion and add to 50.0mL of 0.9% NaCl solution. Mix thoroughly. Filter sterilize. Warm to 45°–50°C. Preparation of Medium: Add components—except. 50°–55°C. Aseptically add 100.0mL of sterile egg yolk emulsion, 50%. Mix thor- oughly. Pour into sterile Petri dishes in 15.0mL volumes. Use: For the cultivation of a wide variety of anaerobic bacteria to form emulsion. Measure 50.0mL of egg yolk emulsion and add to 50.0mL of 0.9% NaCl solution. Mix thoroughly. Filter sterilize. Warm to 45°–50°C. Preparation of Medium: Add components, except