Wilkins-Chalgren Anaerobe Agar with GN Supplement 1905 K 2 HPO 4 0.5g MgSO 4 ·7H 2 O 0.25g Na 2 SO 3 , anhydrous 0.05g pH 7.2 ± 0.2 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 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 Xanthomonas albilineans and other Xanthomonas species. Wilkins-Chalgren Agar Composition per liter: Agar 15.0g Gelatin peptone 10.0g Pancreatic digest of casein 10.0g NaCl 5.0g Yeast extract 5.0g Glucose 1.0g L-Arginine 1.0g Sodium pyruvate 1.0g Hemin 5.0mg Vitamin K 1 (menadione) 0.5mg pH 7.1 ± 0.2 at 25°C Source: This medium is available as a premixed powder from BD Di- agnostic Systems . 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. Cool to 50°–55°C. Add antibiotic to be assayed; varying concentrations of antibiotics are used. Mix thoroughly. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of anaerobic bacteria. For standardized antimicrobic susceptibility testing to determine the mini- mum inhibitory concentrations of antimicrobics for anaerobic bacteria. Wilkins-Chalgren Anaerobe Agar Composition per liter: Agar 10.0g Pancreatic digest of casein 10.0g Gelatin peptone 10.0g NaCl 5.0g Yeast extract 5.0g Glucose 1.0g L-Arginine 1.0g Sodium pyruvate 1.0g Hemin 5.0mg Menadione 0.5mg Defibrinated blood 50.0mL Tween™ 80 1.0mL pH 7.1 ± 0.2 at 25°C Source: This medium is available as a premixed powder from Oxoid Unipath. Preparation of Medium: Add components, except defibrinated blood, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°– 55°C. Aseptically add 50.0mL of defibrinated blood. Mix thoroughly. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation of nonsporulating anaerobes. Wilkins-Chalgren Anaerobe Agar with Cysteine Composition per liter: Agar 10.0g Pancreatic digest of casein 10.0g Gelatin peptone 10.0g NaCl 5.0g Yeast extract 5.0g Glucose 1.0g L-Arginine 1.0g Sodium pyruvate 1.0g L-Cysteine·HCl 0.3g Hemin 5.0mg Menadione 0.5mg Defibrinated blood 50.0mL Tween™ 80 1.0mL pH 7.1 ± 0.2 at 25°C Preparation of Medium: Add components, except defibrinated blood, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°– 55°C. Aseptically add 50.0mL of defibrinated blood. Mix thoroughly. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation of Agromonas oligotrophica, Falcivibrio gran- dis, and Falcivibrio vaginalis. Wilkins-Chalgren Anaerobe Agar with GN Supplement Composition per liter: Agar 10.0g Pancreatic digest of casein 10.0g Gelatin peptone 10.0g NaCl 5.0g Yeast extract 5.0g Glucose 1.0g L-Arginine 1.0g Sodium pyruvate 1.0g Hemin 5.0mg Menadione 0.5mg Defibrinated blood 50.0mL GN anaerobe selective supplement 20.0mL pH 7.1 ± 0.2 at 25°C Source: This medium is available as a premixed powder from Oxoid Unipath. GN Anaerobe Selective Supplement Composition per 20.0mL: Nalidixic acid 10.0mg Hemin 5.0mg Sodium succinate 2.5mg Vancomycin 2.5mg Menadione 0.5mg Preparation of GN Anaerobe Selective Supplement: Add components to distilled/deionized water and bring volume to 20.0mL. Mix thoroughly. Filter sterilize. © 2010 by Taylor and Francis Group, LLC 1906 Wilkins-Chalgren Anaerobe Agar with NS Supplement Preparation of Medium: Add components, except defibrinated blood and GN anaerobe selective supplement, to distilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Aseptically add 20.0mL of GN anaerobe selective supplement and 50.0mL of defibrinated blood. Bring volume to 1.0L with distilled/deionized water. Mix thor- oughly. Pour into sterile Petri dishes or leave in tubes. Use: For the selective isolation of Gram-negative anaerobes. Wilkins-Chalgren Anaerobe Agar with NS Supplement Composition per liter: Agar 10.0g Pancreatic digest of casein 10.0g Gelatin peptone 10.0g NaCl 5.0g Yeast extract 5.0g Glucose 1.0g L-Arginine 1.0g Sodium pyruvate 1.0g Hemin 5.0mg Menadione 0.5mg Defibrinated blood 50.0mL NS anaerobe selective supplement 20.0 mL Tween™ 80 1.0mL pH 7.1 ± 0.2 at 25°C Source: This medium is available as a premixed powder from Oxoid Unipath. NS Anaerobe Selective Supplement: Composition per 20.0mL: Sodium pyruvate 1.0g Nalidixic acid 0.01g Hemin 5.0mg Menadione 0.5mg Preparation of NS Anaerobe Selective Supplement: Add components to distilled/deionized water and bring volume to 20.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except defibrinated blood and NS anaerobe selective supplement, to distilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Aseptically add 20.0mL of NS anaerobe selective supplement and 50.0mL of defibrinated blood. Bring volume to 1.0L with distilled/deionized water. Mix thor- oughly. Pour into sterile Petri dishes or leave in tubes. Use: For the selective isolation of nonsporulating anaerobes. Wilkins-Chalgren Anaerobe Broth (Anaerobe Broth, MIC) Composition per liter: Pancreatic digest of casein 10.0g Gelatin peptone 10.0g NaCl 5.0g Yeast extract 5.0g Glucose 1.0g L-Arginine 1.0g Sodium pyruvate 1.0g Hemin 5.0mg Menadione 0.5mg pH 7.1 ± 0.2 at 25°C Source: This medium is available as a premixed powder from BD Di- agnostic Systems and Oxoid Unipath. 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 antimicrobial susceptibility (MIC) testing of anaerobic bacteria. Wilkins-Chalgren Anaerobe Medium with Yeast Extract Composition per liter: Yeast extract 20.0g Pancreatic digest of casein 10.0g Gelatin peptone 10.0g NaCl 5.0g Yeast extract 5.0g Glucose 1.0g L-Arginine 1.0g Sodium pyruvate 1.0g Hemin 5.0mg Menadione 0.5mg pH 7.1 ± 0.2 at 25°C 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 Selenomonas acidaminovorans. Wilkins-Chalgren Anaerobic HiVeg Agar Base with Blood Composition per liter: Agar 10.0g Plant hydrolysate 10.0g Plant peptone 10.0g NaCl 5.0g Yeast extract 5.0g L-Arginine 1.0g Glucose 1.0g Sodium pyruvate 1.0g Fe 4 (P 2 O 7 ) 3 ·H 2 O 5.0mg Menadione 0.5mg Defibrinated blood 50.0mL pH 7.1 ± 0.2 at 25°C Source: This medium, without blood, is available as a premixed pow- der from HiMedia. Preparation of Medium: Add components, except defibrinated blood, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°– 55°C. Aseptically add 50.0mL of defibrinated blood. Mix thoroughly. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation of nonsporulating anaerobes. For the cultivation and maintenance of anaerobic bacteria. For standardized antimicrobic susceptibility testing to determine the minimum inhibitory concentra- tions of antimicrobics for anaerobic bacteria. © 2010 by Taylor and Francis Group, LLC Wilson Blair Base 1907 Wilkins-Chalgren Anaerobic HiVeg Agar Base with Blood and Nonspore Anaerobic Supplement Composition per liter: Agar 10.0g Plant hydrolysate 10.0g Plant peptone 10.0g NaCl 5.0g Yeast extract 5.0g L-Arginine 1.0g Glucose 1.0g Sodium pyruvate 1.0g Fe 4 (P 2 O 7 ) 3 ·H 2 O 5.0mg Menadione 0.5mg Defibrinated blood 50.0mL Nonspore anaerobic supplement pH 7.1 ± 0.2 at 25°C Source: This medium, without blood or nonspore anaerobic supple- ment, is available as a premixed powder from HiMedia. Nonspore Anaerobic Supplement: Composition per 10.0mL: Sodium pyruvate 1.0g Nalidixic acid 10.0mg Ferric pyrophosphate, soluble 5.0mg Menadione 0.5mg Preparation of Nonspore Anaerobic Supplement: Add com- ponents to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except defibrinated blood and nonspore anaerobic supplement, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Aseptically add 50.0mL of de- fibrinated blood and 10.0mL nonspore anaerobic supplement. Mix thoroughly. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation of nonsporulating anaerobes. For the cultivation and maintenance of anaerobic bacteria. For standardized antimicrobic susceptibility testing to determine the minimum inhibitory concentra- tions of antimicrobics for anaerobic bacteria. Wilkins-Chalgren Anaerobic HiVeg Broth Base with Blood Composition per liter: Plant hydrolysate 10.0g Plant peptone 10.0g Yeast extract 5.0g NaCl 5.0g Sodium pyruvate 1.0g Glucose 1.0g L-Arginine 1.0g Fe 4 (P 2 O 7 ) 3 ·H 2 O 5.0mg Menadione 0.5mg Defibrinated blood 50.0mL pH 7.1 ± 0.2 at 25°C Source: This medium, without blood, is available as a premixed pow- der from HiMedia. Preparation of Medium: Add components, except defibrinated blood, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°– 55°C. Aseptically add 50.0mL of defibrinated blood. Mix thoroughly. Aseptically distribute into tubes or leave in flasks. Use: For the cultivation of nonsporulating anaerobes. For the cultivation and maintenance of anaerobic bacteria. For standardized antimicrobic susceptibility testing to determine the minimum inhibitory concentra- tions of antimicrobics for anaerobic bacteria. Wilson and Blair’s Medium Composition per 165.0mL: Nutrient agar solution 100.0mL Solution B 45.0mL Solution A 20.0mL pH 6.8 ± 0.2 at 25°C Nutrient Agar Solution: Composition per 100.0mL: Agar 1.95g Pancreatic digest of gelatin 0.65g Beef extract 0.39g Preparation of Nutrient Agar Solution: Aseptically add compo- nents to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Solution A: Composition per liter: Na 2 HPO 4 100.0g Na 2 SO 3 100.0g Glucose 50.0g Bismuth ammonium citrate 30.0g Preparation of Solution A: Aseptically add 30.0g of bismuth am- monium citrate to 250.0mL of boiling distilled/deionized water. Asep- tically add Na 2 SO 3 to 500.0mL of boiling distilled/deionized water. Mix the two solutions. Add the Na 2 HPO 4 to the boiling mixture. Cool to 45°C. In a separate flask, aseptically add glucose to 250.0mL of ster- ile distilled/deionized water. Mix thoroughly. Aseptically add the glu- cose solution to the other cooled solution. Solution B: Composition per 225.0mL: Ferric citrate 2.0g Brilliant Green 0.55g Preparation of Solution B: Aseptically add 2.0g of ferric citrate to 200.0mL of sterile distilled/deionized water. Mix thoroughly. Asepti- cally add Brilliant Green to 25.0mL of sterile distilled/deionized water. Mix thoroughly. Aseptically combine the two solutions. Preparation of Medium: Aseptically combine 100.0mL of nutrient agar solution, 20.0mL of solution A, and 45.0mL of solution B. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation of Clostridium welchii. Wilson Blair Base Composition per liter: Agar 30.0g Proteose peptone No. 3 10.0g Glucose 10.0g Beef extract 5.0g NaCl 5.0g © 2010 by Taylor and Francis Group, LLC 1908 Wilson Blair HiVeg Agar with Brilliant Green and Selective Reagent Selective reagent 70.0mL Brilliant Green (1% solution) 4.0mL pH 7.3 ± 0.2 at 25°C Selective Reagent: Composition per 320.2mL: Solution 1 100.0mL Solution 2 100.0mL Solution 3 100.0mL Solution 4 20.2mL Preparation of Selective Reagent: Combine 100.0mL of solution 1, 100.0mL of solution 2, 100.0mL of solution 3, and 20.2mL of solu- tion 4. Mix thoroughly. Gently heat to boiling until a slate-grey color develops. Cool to 50°C. Solution 1: Composition per 100.0mL: NaHSO 3 40.0g Preparation of Solution 1: Add NaHSO 3 to 100.0mL of distilled/ deionized water. Mix thoroughly. Solution 2: Composition per 100.0mL: NaH 2 PO 4 21.0g Preparation of Solution 2: Add NaH 2 PO 4 to 100.0mL of distilled/ deionized water. Mix thoroughly. Solution 3: Composition per 100.0mL: Bismuth ammonium citrate 12.5g Preparation of Solution 3: Add bismuth ammonium citrate to 100.0mL of distilled/deionized water. Mix thoroughly. Solution 4: Composition per 20.2mL: FeSO 4 0.96g Preparation of Solution 4: Add FeSO 4 to 20.0mL of distilled/de- ionized water. Add 0.2mL of concentrated HCl. Mix thoroughly. Preparation of Medium: Add components, except selective re- agent and Brilliant Green solution, to distilled/deionized water and bring volume to 976.0mL. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Aseptically add selective reagent and Brilliant Green solution. Mix thoroughly. Pour into sterile Petri dishes or leave in tubes. Use: For the isolation and cultivation of Salmonella, especially Salmo- nella typhi. Wilson Blair HiVeg Agar with Brilliant Green and Selective Reagent Composition per liter: Agar 20.0g Plant peptone 10.0g Bismuth sulfite indicator 8.0g Glucose 5.0g Plant extract 5.0g Na 2 HPO 4 4.0g FeSO 4 0.3g Brilliant Green 0.025g Selective reagent 70.0mL pH 7.3 ± 0.2 at 25°C Source: This medium, without selective reagent, is available as a pre- mixed powder from HiMedia. Selective Reagent: Composition per 320.2mL: Solution 1 100.0mL Solution 2 100.0mL Solution 3 100.0mL Solution 4 20.2mL Preparation of Selective Reagent: Combine 100.0mL of solution 1, 100.0mL of solution 2, 100.0mL of solution 3, and 20.2mL of solu- tion 4. Mix thoroughly. Gently heat to boiling until a slate-grey color develops. Cool to 50°C. Solution 1: Composition per 100.0mL: NaHSO 3 40.0g Preparation of Solution 1: Add NaHSO 3 to 100.0mL of distilled/ deionized water. Mix thoroughly. Solution 2: Composition per 100.0mL: NaH 2 PO 4 21.0g Preparation of Solution 2: Add NaH 2 PO 4 to 100.0mL of distilled/ deionized water. Mix thoroughly. Solution 3: Composition per 100.0mL: Bismuth ammonium citrate 12.5g Preparation of Solution 3: Add bismuth ammonium citrate to 100.0mL of distilled/deionized water. Mix thoroughly. Solution 4: Composition per 20.2mL: FeSO 4 0.96g Preparation of Solution 4: Add FeSO 4 to 20.0mL of distilled/de- ionized water. Add 0.2mL of concentrated HCl. Mix thoroughly. Preparation of Medium: Add components, except selective re- agent and Brilliant Green solution, to distilled/deionized water and bring volume to 976.0mL. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Aseptically add selective reagent and Brilliant Green solution. Mix thoroughly. Pour into sterile Petri dishes or leave in tubes. Use: For the isolation and cultivation of Salmonella, especially Salmo- nella typhi. Wilson Blair HiVeg Agar Base with Selective Reagent and Brilliant Green Composition per liter: Agar 30.0g Glucose 10.0g Plant special peptone 10.0g Plant extract 5.0g NaCl 5.0g Selective reagent 70.0mL Brilliant Green (1% solution) 4.0mL pH 7.3 ± 0.2 at 25°C Source: This medium, without selective reagent or Brilliant Green, is available as a premixed powder from HiMedia. © 2010 by Taylor and Francis Group, LLC Winogradsky’s N-Free Medium 1909 Selective Reagent: Composition per 100.0mL: Solution 1 100.0mL Solution 2 100.0mL Solution 3 100.0mL Solution 4 20.2mL Preparation of Selective Reagent: Combine 100.0mL of solution 1, 100.0mL of solution 2, 100.0mL of solution 3, and 20.2mL of solu- tion 4. Mix thoroughly. Gently heat to boiling until a slate-grey color develops. Cool to 50°C. Solution 1: Composition per 100.0mL: NaHSO 3 40.0g Preparation of Solution 1: Add NaHSO 3 to 100.0mL of distilled/ deionized water. Mix thoroughly. Solution 2: Composition per 100.0mL: NaH 2 PO 4 21.0g Preparation of Solution 2: Add NaH 2 PO 4 to 100.0mL of distilled/ deionized water. Mix thoroughly. Solution 3: Composition per 100.0mL: Bismuth ammonium citrate 12.5g Preparation of Solution 3: Add bismuth ammonium citrate to 100.0mL of distilled/deionized water. Mix thoroughly. Solution 4: Composition per 20.2mL: FeSO 4 0.96g Preparation of Solution 4: Add FeSO 4 to 20.0mL of distilled/de- ionized water. Add 0.2mL of concentrated HCl. Mix thoroughly. Preparation of Medium: Add components, except selective re- agent and Brilliant Green solution, to distilled/deionized water and bring volume to 976.0mL. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Aseptically add selective reagent and Brilliant Green solution. Mix thoroughly. Pour into sterile Petri dishes or leave in tubes. Use: For the isolation and cultivation of Salmonella, especially Salmo- nella typhi. Winge Agar Composition per liter: Glucose 20.0g Agar 15.0g Yeast extract 3.0g pH 7.2 ± 0.2 at 25°C Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.2. Gently heat and bring to boiling. Distribute into tubes or flasks. Auto- clave 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 Candida albicans, Can- dida kefyr, Candida tropicalis, Citeromyces matritensis, Cryptococcus albidus, Neurospora crassa, Octosporomyces octosporus, Neurospora sitophila, and Saccharomyces species. Winge Melibiose Agar Composition per liter: Agar 15.0g Melibiose 10.0g Yeast extract 3.0g pH 7.2 ± 0.2 at 25°C Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.2. Gently heat and bring to boiling. Distribute into tubes or flasks. Auto- clave 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 Saccharomyces cerevi- siae. Winogradsky’s Medium, Modified Composition per liter: CaCO 3 5.0g (NH 4 ) 2 SO 4 1.0g K 2 HPO 4 1.0g NaCl 1.0g MgSO 4 ·7H 2 O 0.5g FeSO 4 0.4g Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat until dis- solved. Do not autoclave. Distribute into tubes or flasks. Swirl flask while dispensing to suspend precipitate. Use: For the cultivation of nitrifying bacteria. Winogradsky’s N-Free Medium Composition per liter: Agar 20.0g CaCO 3 5.0mg Sugar solution 100.0mL Concentrated salt solution 5.0mL pH 7.2 ± 0.2 at 25°C Sugar Solution: Composition per 100.0mL: Sucrose or glucose 10.0g Preparation of Sugar Solution: Add sucrose or glucose to 100.0mL of distilled/deionized water. Mix thoroughly. Autoclave for 10 min at 10 psi pressure–115°C. Cool to 50°C. Concentrated Salt Solution: Composition per liter: KH 2 PO 4 50.0g MgSO 4 ·7H 2 O 25.0g NaCl 25.0g FeSO 4 ·7H 2 O 1.0g MnSO 4 ·4H 2 O 1.0g Na 2 MoO 4 ·4H 2 O 1.0g Preparation of Concentrated Salt Solution: Add components to tap water and bring volume to 1.0L. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except sugar solution, to distilled/deionized water and bring volume to 900.0mL. Mix thor- oughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Asepti- cally add sugar solution. Adjust pH to 7.2. Mix thoroughly. Pour into sterile Petri dishes or leave in tubes. © 2010 by Taylor and Francis Group, LLC 1910 Winogradsky’s Nitrite Medium Use: For the cultivation and maintenance of Azomonas insignis. Winogradsky’s Nitrite Medium Composition per liter: Agar 15.0g NaNO 2 2.0g Na 2 CO 3 , anhydrous 1.0g K 2 HPO 4 0.5g 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 selective isolation and cultivation of Nocardia species and Rhodococcus species. WL Differential Agar (Wallerstein Laboratory Differential Agar) Composition per liter: Glucose 50.0g Agar 20.0g Pancreatic digest of casein 5.0g Yeast extract 4.0g KH 2 PO 4 0.55g KCl 0.425g CaCl 2 ·2H 2 O 0.125g MgSO 4 ·7H 2 O 0.125g Bromcresol Green 0.022g Actidione ® (cycloheximide) 4.0mg FeCl 3 2.5mg MnSO 4 ·4H 2 O 2.5mg pH 5.5 ± 0.2 at 25°C Caution: Cycloheximide is toxic. Avoid skin contact or aerosol for- mation and inhalation. Source: This medium is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat with mix- ing 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 differential cultivation of bacteria from industrial fermen- tation processes. Growth of yeasts and molds is inhibited. WL Differential HiVeg Agar Composition per liter: Glucose 50.0g Agar 20.0g Plant hydrolysate 5.0g Yeast extract 4.0g KH 2 PO 4 0.55g KCl 0.425g MgSO 4 0.125g CaCl 2 0.125g Bromcresol Green 0.022g Cycloheximide 4.0mg FeCl 3 2.5mg MnSO 4 2.5mg pH 5.5 ± 0.2 at 25°C Source: This medium is available as a premixed powder from Hi- Media. Caution: Cycloheximide is toxic. Avoid skin contact or aerosol for- mation and inhalation. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat with mix- ing 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 differential cultivation of bacteria from industrial fermen- tation processes. Growth of yeasts and molds is inhibited. For the selective isolation and enumeration of bacteria encountered in brewer- ies and industrial fermentations. WL Differential HiVeg Broth Composition per liter: Glucose 50.0g Plant hydrolysate 5.0g Yeast extract 4.0g KH 2 PO 4 0.55g KCl 0.425g MgSO 4 0.125g CaCl 2 0.125g Bromcresol Green 0.022g Cycloheximide 4.0mg FeCl 3 2.5mg MnSO 4 2.5mg pH 5.5 ± 0.2 at 25°C Source: This medium is available as a premixed powder from Hi- Media. Caution: Cycloheximide is toxic. Avoid skin contact or aerosol for- mation and inhalation. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat with mix- ing and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the differential cultivation of bacteria from industrial fermen- tation processes. Growth of yeasts and molds is inhibited. For the selective isolation and enumeration of bacteria encountered in brewer- ies and industrial fermentations. WL Differential Medium (Wallerstein Laboratory Differential Medium) Composition per liter: Glucose 50.0g Agar 20.0g Pancreatic digest of casein 5.0g Yeast extract 4.0g KH 2 PO 4 0.55g KCl 0.425g CaCl 2 ·2H 2 O 0.125g MgSO 4 ·7H 2 O 0.125g Bromcresol Green 0.022g Actidione ® (cycloheximide) 4.0mg FeCl 3 2.5mg MnSO 4 ·4H 2 O 2.5mg pH 5.5 ± 0.2 at 25°C © 2010 by Taylor and Francis Group, LLC WL Nutrient Agar 1911 Source: This medium is available as a premixed powder from BD Di- agnostic Systems. Caution: Cycloheximide is toxic. Avoid skin contact or aerosol for- mation and inhalation. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat with mix- ing 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 differential cultivation of bacteria from industrial fermen- tation processes. Growth of yeasts and molds is inhibited. WL Differential Medium (Wallerstein Laboratory Differential Medium) Composition per liter: Glucose 50.0g Agar 20.0g Pancreatic digest of casein 5.0g Yeast extract 4.0g KH 2 PO 4 0.55g KCl 0.425g CaCl 2 ·2H 2 O 0.125g MgSO 4 ·7H 2 O 0.125g Bromcresol Green 0.022g Actidione ® (cycloheximide) 4.0mg FeCl 3 2.5mg MnSO 4 ·4H 2 O 2.5mg Na 2 CO 3 (1% solution) 30.0mL pH 6.5 ± 0.2 at 25°C Source: This medium is available as a premixed powder from BD Di- agnostic Systems. Caution: Cycloheximide is toxic. Avoid skin contact or aerosol for- mation and inhalation. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat with mix- ing 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 differential cultivation of bacteria from industrial fermen- tation processes. Growth of yeasts and molds is inhibited. WL Medium with Tomato Juice (Wallerstein Laboratory Medium with Tomato Juice) Composition per liter: Glucose 50.0g Pancreatic digest of casein 5.0g Yeast extract 4.0g KH 2 PO 4 0.55g KCl 0.425g CaCl 2 0.125g MgSO 4 ·7H 2 O 0.125g Bromcresol Green 0.022g FeCl 3 2.5mg MnSO 4 ·4H 2 O 2.5mg Tomato juice, canned, clarified 400.0mL pH 5.5 ± 0.2 at 25°C 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 microorganisms from alcoholic mash. WL Nutrient Agar (Wallerstein Laboratory Nutrient Agar) Composition per liter: Glucose 50.0g Agar 20.0g Pancreatic digest of casein 5.0g Yeast extract 4.0g KH 2 PO 4 0.55g KCl 0.425g CaCl 2 ·2H 2 O 0.125g MgSO 4 ·7H 2 O 0.125g Bromcresol Green 0.022g FeCl 3 2.5mg MnSO 4 ·4H 2 O 2.5mg pH 5.5 ± 0.2 at 25°C Source: This medium is available as a premixed powder from BD Di- agnostic Systems and Oxoid Unipath. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat with mix- ing 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 detection of bacteria and yeasts in industrial fermentation processes, particularly from beer processing. WL Nutrient Agar (Wallerstein Laboratory Nutrient Agar) Composition per liter: Glucose 50.0g Agar 20.0g Pancreatic digest of casein 5.0g Yeast extract 4.0g KH 2 PO 4 0.55g KCl 0.425g CaCl 2 ·2H 2 O 0.125g MgSO 4 ·7H 2 O 0.125g Bromcresol Green 0.022g FeCl 3 2.5mg MnSO 4 ·4H 2 O 2.5mg Na 2 CO 3 (1% solution) 30.0mL pH 6.5 ± 0.2 at 25°C Source: This medium is available as a premixed powder from BD Di- agnostic Systems and Oxoid Unipath. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat with mix- ing 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 detection of bacteria and yeasts from industrial fermenta- tion processes, particularly from beer processing. © 2010 by Taylor and Francis Group, LLC 1912 WL Nutrient Broth WL Nutrient Broth (Wallerstein Laboratory Nutrient Broth) Composition per liter: Glucose 50.0g Pancreatic digest of casein 5.0g Yeast extract 4.0g KH 2 PO 4 0.55g KCl 0.425g CaCl 2 0.125g MgSO 4 ·7H 2 O 0.125g Bromcresol Green 0.022g FeCl 3 2.5mg MnSO 4 ·4H 2 O 2.5mg pH 5.5 ± 0.2 at 25°C Source: This medium is available as a premixed powder from BD Di- agnostic Systems and Oxoid Unipath. 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 yeasts, molds, and bacteria found in brew- ing and other industrial fermentation processes. WL Nutrient Broth (Wallerstein Laboratory Nutrient Broth) Composition per liter: Glucose 50.0g Pancreatic digest of casein 5.0g Yeast extract 4.0g KH 2 PO 4 0.55g KCl 0.425g CaCl 2 0.125g MgSO 4 ·7H 2 O 0.125g Bromcresol Green 0.022g FeCl 3 2.5mg MnSO 4 ·4H 2 O 2.5mg Na 2 CO 3 (1% solution) 30.0mL pH 6.5 ± 0.2 at 25°C Source: This medium is available as a premixed powder from BD Di- agnostic Systems and Oxoid Unipath. 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 yeasts, molds, and bacteria found in brew- ing and other industrial fermentation processes. WL Nutrient Broth (Wallerstein Laboratory Nutrient Broth) Composition per liter: Glucose 50.0g Pancreatic digest of casein 5.0g Yeast extract 4.0g FeCl 3 2.5g MnSO 4 ·4H 2 O 2.5g KH 2 PO 4 0.55g KCl 0.425g CaCl 2 ·2H 2 O 0.125g MgSO 4 ·7H 2 O 0.125g Bromcresol Green 0.022g pH 5.5 ± 0.2 at 25°C 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 control of brewing and other fermentation processes. WL Nutrient HiVeg Broth Composition per liter: Glucose 50.0g Plant hydrolysate 5.0g Yeast extract 4.0g KH 2 PO 4 0.55g KCl 0.425g MgSO 4 0.125g CaCl 2 0.125g Bromcresol green 0.022g FeCl 3 2.5mg MnSO 4 2.5mg pH 5.5 ± 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. Gently heat with mix- ing and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the differential cultivation of bacteria from industrial fermen- tation processes. Growth of yeasts and molds is inhibited. For the selective isolation and enumeration of bacteria encountered in brewer- ies and industrial fermentations. WL Nutrient HiVeg Medium Composition per liter: Glucose 50.0g Agar 20.0g Plant hydrolysate 5.0g Yeast extract 4.0g KH 2 PO 4 0.55g KCl 0.425g CaCl 2 0.125g MgSO 4 0.125g Bromcresol green 0.022g FeCl 3 2.5mg MnSO 4 2.5mg pH 5.5 ± 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. Gently heat with mix- ing and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the differential cultivation of bacteria from industrial fermen- tation processes. Growth of yeasts and molds is inhibited. For the selective isolation and enumeration of bacteria encountered in brewer- ies and industrial fermentations. WMC Medium Composition per 1010.0mL: NaHCO 3 5.0g Sodium acetate 1.0g L-Cysteine 0.5g © 2010 by Taylor and Francis Group, LLC Wolinella succinogenes Medium 1913 Resazurin 1.0mg Mineral salt solution 2xW 500.0mL LIP solution 50.0mL TYC solution 50.0mL Trace elements solution 10.0mL Na 2 S·9H 2 O solution 10.0mL pH 7.2 ± 0.2 at 25°C Mineral Solution 2xW: Composition per liter: NaCl 40.0g MgCl 2 ·6H 2 O 5.6g MgSO 4 ·7H 2 O 0.7g KCl 0.68g NH 4 Cl 0.5g CaCl 2 ·2H 2 O 0.28g K 2 HPO 4 .0.28g Preparation of Mineral Solution 2xW: Add components to dis- tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize. Store at room temperature in the dark. LIP Solution: Composition per liter: L-Isoleucine 10.0g L-Leucine 5.0g Pantothenate 0.1g Preparation of LIP Solution: Add components to distilled/deion- ized water and bring volume to 1.0L. Mix thoroughly. May be stored unsterilized at −20°C. TYC Solution: Composition per liter: Casamino acids 100.0g Yeast extract 50.0g L-Tryptophan 1.0g Preparation of TYC Solution: Add components to distilled/de- ionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Trace Elements Solution: Composition per liter: MgSO 4 ·7H 2 O 3.0g Nitrilotriacetic acid 1.5g NaCl 1.0g MnSO 4 ·2H 2 O 0.5g CoSO 4 ·7H 2 O 0.18g ZnSO 4 ·7H 2 O 0.18g FeSO 4 ·7H 2 O 0.1g CaCl 2 ·2H 2 O 0.1g KAI(SO 4 ) 2 ·12H 2 O 0.02g CuSO 4 ·5H 2 O 0.01g H 3 BO 3 0.01g Na 2 MoO 4 ·2H 2 O 0.01g NiCl 2 ·6H 2 O 0.025g Na 2 SeO 3 ·5H 2 O 0.3mg Preparation of Trace Elements Solution: Add nitrilotriacetic acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with KOH. Add remaining components. Add distilled/deionized water to 1.0L. Na 2 S·9H 2 O Solution: Composition per 10.0mL: Na 2 S·9H 2 O 0.5g Preparation of Na 2 S·9H 2 O Solution: Add Na 2 S·9H 2 O to dis- tilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: Prepare and dispense medium under 80% H 2 + 20% CO 2 . Add components, except TYC solution and Na 2 S·9H 2 O solution, to distilled/deionized water and bring volume to 940.0mL. Mix thoroughly. Sparge with 80% H 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add 50.0mL of sterile TYC solution and 10.0mL of sterile Na 2 S·9H 2 O so- lution. Mix thoroughly. Use: For the cultivation and maintenance of Methanococcus voltae. Wolin-Bevis Medium Composition per liter: Agar 20.0g (NH 4 ) 2 SO 4 1.0g KH 2 HPO 4 1.0g Glucose 0.25g L-Histidine·HCl 0.25g Tween™ 80 (polysorbate 80) 3.0mL pH 5.4 ± 0.2 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 or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the enhanced production of chlamydospores by Candida albicans. Wolinella succinogenes Medium Composition per liter: K 2 HPO 4 5.0g Fumaric acid 3.0g Sodium formate 3.0g (NH 4 ) 2 SO 4 1.0g Yeast extract 1.0g MgCl 2 ·6H 2 O 0.2g FeSO 4 ·7H 2 O 0.02g Resazurin 1.0mg Sodium thioglycolate solution 10.0mL pH 7.0 ± 0.2 at 25°C Sodium Thioglycolate Solution: Composition per 10.0mL: Sodium thioglycolate 0.5g Preparation of Sodium Thioglycolate Solution: Add sodium thioglycolate to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: Add components, except sodium thiogly- colate solution, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure– 121°C. Aseptically add 10.0mL of sterile sodium thioglycolate solu- tion. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation and maintenance of Wolinella succinogenes. © 2010 by Taylor and Francis Group, LLC 1914 Woods and Welton Agar Woods and Welton Agar Composition per liter: NaCl 23.4g Casein hydrolysate 17.0g Agar 15.0g Glycerol 10.0g Pancreatic digest of gelatin 5.0g Glucose 5.0g Papaic digest of soybean meal 3.0g Beef extract 3.0g Yeast extract 2.0g Casamino acids, vitamin free 0.5g Pancreatic digest of casein 0.5g Na 2 SO 3 0.1g pH 7.6 ± 0.2 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. Adjust pH to 7.6. 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 Vibrio alginolyticus. Worfel-Ferguson Agar Composition per liter: Sucrose 20.0g Agar 15.0g NaCl 2.0g Yeast extract 2.0g K 2 SO 4 1.0g MgSO 4 ·7H 2 O 0.25g pH 6.5 ± 0.2 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 or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the detection of capsule production by Klebsiella. For sero- logical detection of the Neufeld (Quellung) reaction. Wort Agar Composition per liter: Agar 15.0g Malt extract 15.0g Maltose 12.75g Dextrin 2.75g Glycerol 2.35g K 2 HPO 4 1.0g NH 4 Cl 1.0g Pancreatic digest of gelatin 0.78g pH 4.8 ± 0.2 at 25°C Source: This medium is available as a premixed powder from BD Di- agnostic Systems and Oxoid Unipath. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Boil for 1 min with mixing. Distribute into tubes or flasks. Au- toclave for 15 min at 15 psi pressure–121°C. Do not overheat, as this will result in hydrolysis of the agar. An additional 5.0g of agar can be used to make a firmer agar. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and enumeration of yeasts. The low pH of the agar selectively inhibits bacterial growth. Wort Agar Composition per liter: Agar 25.0g Yeast extract 1.0g Wort solution 1.0L Wort Solution: Composition per liter: Malt extract 110.0g Preparation of Wort Solution: Add malt extract to distilled/deion- ized water and bring volume to 1.0L. Mix thoroughly. Preparation of Medium: Combine components. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Auto- clave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and mainteneance of a wide variety of yeasts and filamentous fungi. Wort Agar 6°Brix Composition per liter: Malt 250.0g Agar 20.0g Preparation of Medium: Add 250.0g of ground malt to tap water and bring volume to 1.0L. Mix thoroughly. Gently heat to 55°–60°C. Maintain at 55°–60°C for 1.5–2.0 hr while mixing regularly. Elevate temperature to 80°C and maintain for 10 min with thorough mixing. Cool the wort to 25°C and filter through a linen bag. Adjust the con- centration of sugars to 6° Brix (specific gravity of 1.024 at 24°C). Add 20.0g of agar. Gently heat and bring to boiling. Mix thoroughly. Dis- tribute into tubes or flasks. Autoclave for 30 min at 3 psi pressure– 105°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Rhodococcus erythropo- lis. Wort Broth Composition per liter: Yeast extract 1.0g Wort solution 1.0L Wort Solution: Composition per liter: Malt extract 110.0g Preparation of Wort Solution: Add malt extract to distilled/deion- ized water and bring volume to 1000.0mL. Mix thoroughly. 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 cultivation of a wide variety of yeasts and filamentous fungi. Wort HiVeg Agar Composition per liter: Agar 15.0g Malt extract 15.0g Maltose 12.75g Dextrin 2.75g © 2010 by Taylor and Francis Group, LLC . 3 100.0mL Solution 4 20.2mL Preparation of Selective Reagent: Combine 100.0mL of solution 1, 100.0mL of solution 2, 100.0mL of solution 3, and 20.2mL of solu- tion 4. Mix thoroughly. Gently heat. 3 100.0mL Solution 4 20.2mL Preparation of Selective Reagent: Combine 100.0mL of solution 1, 100.0mL of solution 2, 100.0mL of solution 3, and 20.2mL of solu- tion 4. Mix thoroughly. Gently heat. 3 100.0mL Solution 4 20.2mL Preparation of Selective Reagent: Combine 100.0mL of solution 1, 100.0mL of solution 2, 100.0mL of solution 3, and 20.2mL of solu- tion 4. Mix thoroughly. Gently heat