Handbook of Microbiological Media, Fourth Edition part 172 ppt

Thauera aromatica AR-1 Medium 1705 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 Deinococcus species. Thauera aromatica AR-1 Medium (DSMZ Medium 855) Composition per 992.0mL: Solution A 870.0mL Solution C 100.0mL Solution D 10.0mL Solution E (Vitamin solution) 10.0mL Solution B (Trace elements solution SL-10) 1.0mL Selenite-tungstate solution 1.0mL pH 7.2 ± 0.2 at 25°C Solution A: Composition per 870.0mL: Na 2 SO 4 3.0g NaCl 1.0g KNO 3 0.6g KCl 0.5g MgCl 2 ·6H 2 O 0.4g NH 4 Cl 0.3g KH 2 PO 4 0.2g CaCl 2 ·2H 2 O 0.15g Resazurin 1.0mg Preparation of Solution A: Add components to distilled/deionized water and bring volume to 870.0mL. Mix thoroughly. Solution B (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 Solution B (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 thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Solution C: Composition per 100.0mL: NaHCO 3 5.0g Preparation of Solution C: Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Flush with 80% N 2 + 20% CO 2 to remove dissolved oxygen. Solution D: Composition per 10.0mL: Na-benzoate 0.7g Preparation of Solution D: Add Na-benzoate 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. Solution E (Vitamin Solution): Composition per liter: Pyridoxine-HCl 10.0mg Thiamine-HCl·2H 2 O 5.0mg Riboflavin 5.0mg Nicotinic acid 5.0mg D-Ca-pantothenate 5.0mg p-Aminobenzoic acid 5.0mg Lipoic acid 5.0mg Biotin 2.0mg Folic acid 2.0mg Vitamin B 12 0.10mg Solution E (Vitamin Solution): Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Selenite-Tungstate Solution Composition per liter: NaOH 0.5g Na 2 WO 4 ·2H 2 O 4.0mg Na 2 SeO 3 ·5H 2 O 3.0mg Preparation of Selenite-Tungstate Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Preparation of Medium: Gently heat solution A and bring to boiling. Boil solution A for a few minutes. Cool to room temperature. Gas with 80% N 2 + 20% CO 2 gas mixture to reach a pH below 6. Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature. Se- quentially add 1.0mL solution B, 100.0mL solution C, 10.0mL solution D, 10.0mL solution E, and 1.0mL sterile selenite-tungstate solution. Distribute anaerobically under 80% N 2 + 20% CO 2 into ap- propriate vessels. Addition of 10–20mg sodium dithionite per liter from a 5% (w/v) solution, freshly prepared under N 2 and filter-steril- ized, may stimulate growth. Use: For the anaerobic cultivation of Thauera aromatica. Thauera aromatica AR-1 Medium (DSMZ Medium 855) Composition per 892.0mL: Solution A 870.0mL Solution C 10.0mL Solution D (Vitamin solution) 10.0mL Solution B (Trace elements solution SL-10) 1.0mL Selenite-tungstate solution 1.0mL pH 7.2 ± 0.2 at 25°C Solution A: Composition per 870.0mL: NaCl 1.0g Na 2 SO 4 3.0g MgCl 2 ·6H 2 O 0.4g KH 2 PO 4 0.2g NH 4 Cl 0.3g KCl 0.5g CaCl 2 ·2H 2 O 0.15g Resazurin 1.0mg Preparation of Solution A: Add components to distilled/deionized water and bring volume to 870.0mL Mix thoroughly. © 2010 by Taylor and Francis Group, LLC 1706 Thauera aromatica Medium Solution B (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 Solution B (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 thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Solution C: Composition per 10.0mL: Na-benzoate 0.7g Preparation of Solution C: Add Na-benzoate 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. Solution D (Vitamin Solution): Composition per liter: Pyridoxine-HCl 10.0mg Thiamine-HCl·2H 2 O 5.0mg Riboflavin 5.0mg Nicotinic acid 5.0mg D-Ca-pantothenate 5.0mg p-Aminobenzoic acid 5.0mg Lipoic acid 5.0mg Biotin 2.0mg Folic acid 2.0mg Vitamin B 12 0.10mg Solution D (Vitamin Solution): Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Selenite-Tungstate Solution Composition per liter: NaOH 0.5g Na 2 WO 4 ·2H 2 O 4.0mg Na 2 SeO 3 ·5H 2 O 3.0mg Preparation of Selenite-Tungstate Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Preparation of Medium: Adjust pH of solution A to 7.0. Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature. Se- quentially add 1.0mL solution B, 10.0mL solution C, 10.0mL solution D, and 1.0mL sterile selenite-tungstate solution. Adjust pH to 7.2. Aseptically distribute into tubes or flasks. Use: For the aerobic cultivation of Thauera aromatica. Thauera aromatica Medium Composition 1015.0mL: Solution A 500.0mL Solution B 500.0mL Trace elements solution SL-10 10.0mL Vitamin solution 5.0mL pH 7.5 ± 0.2 at 25°C Solution A: Composition per 500.0mL: K 2 HPO 4 5.92g KH 2 PO 4 0.816g Preparation of Solution A: Add components to distilled/deionized water and bring volume to 500.0mL. Mix thoroughly. Adjust pH to 7.5. Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature. Solution B: Composition per 500.0mL: KNO 3 2.0g Sodium benzoate 0.72g NH 4 Cl 0.267g MgSO 4 ·7H 2 O 0.197g CaCl 2 ·2H 2 O 0.025g Preparation of Solution B: Add components to distilled/deionized water and bring volume to 500.0mL. Mix thoroughly. Adjust pH to 7.5. Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature. 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 thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure– 121°C. Cool to room temperature. Vitamin Solution: Composition per liter: Pyridoxine·HCl 10.0mg p-Aminobenzoic acid 5.0mg Lipoic acid 5.0mg Nicotinic acid 5.0mg Riboflavin 5.0mg Thiamine·HCl 5.0mg Calcium DL-pantothenate 5.0mg Biotin 2.0mg Folic acid 2.0mg Vitamin B 12 0.1mg Preparation of Vitamin Solution: Add components to distilled/ deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize. Preparation of Medium: Aseptically combine 500.0mL of sterile solution A, 500.0mL of sterile solution B, 10.0 mL of sterile trace elements solution SL-10, and 5.0 mL of sterile vitamin solution. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation of Thauera aromatica. © 2010 by Taylor and Francis Group, LLC Thayer-Martin HiVeg Medium Base with Hemoglobin and Vitox Supplement 1707 Thauera mechernichi Medium (DSMZ Medium 918) Composition per liter: Na 2 HPO 4 4.20g Na-acetate 2.93g KH 2 PO 4 1.5g NH 4 Cl 0.3g MgSO 4 ·7H 2 O 0.1g Trace elements solution 2.0mL pH 7.1 ± 0.2 at 25°C Trace Elements Solution: Composition per liter: Na 2 -EDTA 50.0g CaCl 2 ·2H 2 O 5.5g MnCl 2 ·4H 2 O 5.06g FeSO 4 ·7H 2 O 5.0g ZnSO 4 ·7H 2 O 2.0g CoCl 2 ·6H 2 O 1.61g CuSO 4 ·5H 2 O 1.57g (NH 4 ) 6 Mo 7 O 24 ·4H 2 O 1.1g Preparation of Trace Elements Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 6.0. 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 Thauera mechernichensis. Thayer-Martin Agar, Modified (MTM II) (Modified Thayer-Martin Agar) Composition per liter: Agar 12.0g Hemoglobin 10.0g Pancreatic digest of casein 7.5g Selected meat peptone 7.5g NaCl 5.0g K 2 HPO 4 4.0g Cornstarch 1.0g KH 2 PO 4 1.0g CNVT inhibitor 10.0mL Supplement solution 10.0mL pH 7.2 ± 0.2 at 25°C CNVT Inhibitor: Composition per 10.0mL: Colistin sulfate 7.5mg Trimethoprim lactate 5.0mg Vancomycin 3.0mg Nystatin 12,500U Preparation of CNVT Inhibitor: Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Supplement Solution: Composition per liter: Glucose 100.0g L-Cysteine·HCl 25.9g L-Glutamine 10.0g L-Cystine 1.1g Adenine 1.0g Nicotinamide adenine dinucleotide 0.25g Vitamin B 12 0.1g Thiamine pyrophosphate 0.1g Guanine·HCl 0.03g Fe(NO 3 ) 3 ·6H 2 O 0.02g p-Aminobenzoic acid 0.013g Thiamine·HCl 3.0mg Source: The supplement solution IsoVitaleX ® enrichment is available from BD Diagnostic Systems. This enrichment may be replaced by supplement VX from BD Diagnostic Systems. Preparation of Supplement Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except CNVT inhibitor and supplement solution, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Gently heat and bring to boiling. Distrib- ute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add 10.0mL of sterile CNVT inhibitor and 10.0mL of sterile supplement solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the isolation of Neisseria species from specimens containing mixed flora of bacteria and fungi. Thayer-Martin HiVeg Medium Base with Hemoglobin and Vitox Supplement Composition per liter: Plant special peptone 23.0g Agar 13.0g NaCl 5.0g Starch 1.0g Hemoglobin solution 250.0mL Vitox supplement 10.0mL pH 7.0 ± 0.2 at 25°C Source: This medium, without hemoglobin or Vitox supplement, is available as a premixed powder from HiMedia. Hemoglobin Solution: Composition per 250.0mL: Hemoglobin 5.0g Preparation of Hemoglobin Solution: Add hemoglobin to distilled/deionized water and bring volume to 250.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Vitox Supplement: Composition per 10.0mL: Glucose 2.0g L-Cysteine·HCl 0.518g L-Glutamine 0.2g L-Cystine 0.022g Adenine sulfate 0.01g Nicotinamide adenine dinucleotide 5.0mg Cocarboxylase 2.0mg Guanine·HCl 0.6mg Fe(NO 3 ) 3 ·6H 2 O 0.4mg p-Aminobenzoic acid 0.26mg Vitamin B 12 0.2mg Thiamine·HCl 0.06mg © 2010 by Taylor and Francis Group, LLC 1708 Thayer-Martin Medium Preparation of Vitox Supplement: Add components to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except hemoblobin and Vitox supplement, to distilled/deionized water and bring volume to 740.0mL. Mix thoroughly. Gently heat until boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add 250.0mL of sterile hemoglobin solution and 10.0mL of sterile Vitox supplement. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the isolation and cultivation of fastidious microorganisms, especially Neisseria species. For the selective isolation of gonococci from pathological specimens. Thayer-Martin Medium Composition per liter: GC agar base 740.0mL Hemoglobin solution 250.0mL Vitox supplement 10.0mL pH 7.3 ± 0.2 at 25°C GC Agar Base: Composition per 740.0mL: Special peptone 15.0g Agar 10.0g NaCl 5.0g K 2 HPO 4 4.0g Cornstarch 1.0g KH 2 PO 4 1.0g pH 7.2 ± 0.2 at 25°C Preparation of GC Agar Base: Add components of GC medium base and the hemoglobin to distilled/deionized water and bring volume to 740.0mL. Mix thoroughly. Gently heat until boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Hemoglobin Solution: Composition per 250.0mL: Hemoglobin 5.0g Preparation of Hemoglobin Solution: Add hemoglobin to distilled/deionized water and bring volume to 250.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Vitox Supplement: Composition per 10.0mL: Glucose 2.0g L-Cysteine·HCl 0.518g L-Glutamine 0.2g L-Cystine 0.022g Adenine sulfate 0.01g Nicotinamide adenine dinucleotide 5.0mg Cocarboxylase 2.0mg Guanine·HCl 0.6mg Fe(NO 3 ) 3 ·6H 2 O 0.4mg p-Aminobenzoic acid 0.26mg Vitamin B 12 0.2mg Thiamine·HCl 0.06mg Preparation of Vitox Supplement: Add components to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: To 740.0mL of cooled sterile GC agar base, aseptically add 250.0mL of sterile hemoglobin solution and 10.0mL of sterile Vitox supplement. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the isolation and cultivation of fastidious microorganisms, especially Neisseria species. Thayer-Martin Medium Composition per liter: Hemoglobin 10.0g GC medium base 980.0mL CNVT inhibitor 10.0mL Supplement B 10.0mL pH 7.3 ± 0.2 at 25°C Source: This medium is available as a prepared medium in tubes from BD Diagnostic Systems. GC Medium Base: Composition per 980.0mL: Proteose peptone No. 3 15.0g Agar 10.0g NaCl 5.0g K 2 HPO 4 4.0g Cornstarch 1.0g KH 2 PO 4 1.0g pH 7.2 ± 0.2 at 25°C Preparation of GC Medium Base: Add components of GC medium base and the hemoglobin to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat until boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. CNVT Inhibitor: Composition per 10.0mL: Colistin sulfate 7.5mg Trimethoprim lactate 5.0mg Vancomycin 3.0mg Nystatin 12,500U Preparation of CNVT Inhibitor: Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: To 980.0mL of cooled sterile GC medium base, aseptically add 10.0mL of sterile CNVT inhibitor and 10.0mL of sterile supplement B. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the isolation and cultivation of fastidious microorganisms, especially Neisseria species. Thayer-Martin Medium, Modified (Modified Thayer-Martin Agar) Composition per liter: GC agar base 720.0mL Hemoglobin solution 250.0mL GC supplement 30.0mL pH 7.3 ± 0.2 at 25°C GC Agar Base: Composition per 720.0mL: Special peptone 15.0g Agar 10.0g NaCl 5.0g K 2 HPO 4 4.0g © 2010 by Taylor and Francis Group, LLC Thayer-Martin Medium, Selective 1709 Cornstarch 1.0g KH 2 PO 4 1.0g pH 7.2 ± 0.2 at 25°C Preparation of GC Agar Base: Add components of GC medium base to distilled/deionized water and bring volume to 720.0mL. Mix thoroughly. Gently heat until boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Hemoglobin Solution: Composition per 250.0mL: Hemoglobin 5.0g Preparation of Hemoglobin Solution: Add hemoglobin to distilled/deionized water and bring volume to 250.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. GC Supplement: Composition per 30.0mL: Yeast autolysate 10.0g Glucose 1.5g NaHCO 3 0.15g Colistin sulfate 7.5mg Trimethoprim lactate 5.0mg Vancomycin 3.0mg Nystatin 12,500U Preparation of GC Supplement: Add components to distilled/deionized water and bring volume to 30.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: To 720.0mL of cooled sterile GC agar base, aseptically add 250.0mL of sterile hemoglobin solution and 30.0mL of sterile GC supplement. Mix thoroughly. Pour into sterile Pe- tri dishes or distribute into sterile tubes. Use: For the selective isolation and cultivation of fastidious microorganisms, especially Neisseria species. Thayer-Martin Medium, Modified (Modified Thayer-Martin Agar) Composition per liter: GC agar base 730.0mL Hemoglobin solution 250.0mL Vitox supplement 10.0mL VCNT antibiotic solution 10.0mL pH 7.3 ± 0.2 at 25°C GC Agar Base: Composition per 730.0mL: Special peptone 15.0g Agar 10.0g NaCl 5.0g K 2 HPO 4 4.0g Cornstarch 1.0g KH 2 PO 4 1.0g pH 7.2 ± 0.2 at 25°C Preparation of GC Agar Base: Add components of GC medium base in to distilled/deionized water and bring volume to 730.0mL. Mix thoroughly. Gently heat until boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Hemoglobin Solution: Composition per 250.0mL: Hemoglobin 5.0g Preparation of Hemoglobin Solution: Add hemoglobin to distilled/deionized water and bring volume to 250.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Vitox Supplement: Composition per 10.0mL: Glucose 2.0g L-Cysteine·HCl 0.518g L-Glutamine 0.2g L-Cystine 0.022g Adenine sulfate 0.01g Nicotinamide adenine dinucleotide 5.0mg Cocarboxylase 2.0mg Guanine·HCl 0.6mg Fe(NO 3 ) 3 ·6H 2 O 0.4mg p-Aminobenzoic acid 0.26mg Vitamin B 12 0.2mg Thiamine·HCl 0.06mg Preparation of Vitox Supplement: Add components to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. VCNT Antibiotic Solution: Composition per 10.0mL: Colistin methane sulfonate 7.5mg Trimethoprim lactate 5.0mg Vancomycin 3.0mg Nystatin 12,500U Preparation of VCNT Antibiotic Solution: Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: To 730.0mL of cooled, sterile GC agar base, aseptically add 250.0mL of sterile hemoglobin solution, 10.0mL of sterile Vitox supplement, and 10.0mL of VCNT antibiotic solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the selective isolation and cultivation of fastidious microorganisms, especially Neisseria species. Thayer-Martin Medium, Selective Composition per liter: GC agar base 730.0mL Hemoglobin solution 250.0mL Vitox supplement 10.0mL VCN antibiotic solution 10.0mL pH 7.3 ± 0.2 at 25°C GC Agar Base: Composition per 730.0mL: Special peptone 15.0g Agar 10.0g NaCl 5.0g K 2 HPO 4 4.0g Cornstarch 1.0g KH 2 PO 4 1.0g pH 7.2 ± 0.2 at 25°C Preparation of GC Agar Base: Add components of GC medium base to distilled/deionized water and bring volume to 730.0mL. Mix thoroughly. Gently heat until boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. © 2010 by Taylor and Francis Group, LLC 1710 Thayer-Martin Selective Agar Hemoglobin Solution: Composition per 250.0mL: Hemoglobin 5.0g Preparation of Hemoglobin Solution: Add hemoglobin to distilled/deionized water and bring volume to 250.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Vitox Supplement: Composition per 10.0mL: Glucose 2.0g L-Cysteine·HCl 0.518g L-Glutamine 0.2g L-Cystine 0.022g Adenine sulfate 0.01g Nicotinamide adenine dinucleotide 5.0mg Cocarboxylase 2.0mg Guanine·HCl 0.6mg Fe(NO 3 ) 3 ·6H 2 O 0.4mg p-Aminobenzoic acid 0.26mg Vitamin B 12 0.2mg Thiamine·HCl 0.06mg Preparation of Vitox Supplement: Add components to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. VCN Antibiotic Solution: Composition per 10.0mL: Colistin methane sulfonate 7.5mg Vancomycin 3.0mg Nystatin 12,500U Preparation of VCN Antibiotic Solution: Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: To 730.0mL of cooled, sterile GC agar base, aseptically add 250.0mL of sterile hemoglobin solution, 10.0mL of sterile Vitox supplement, and 10.0mL of VCN antibiotic solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the selective isolation and cultivation of fastidious microorganisms, especially Neisseria species. Thayer-Martin Selective Agar Composition per liter: Agar 12.0g Hemoglobin 10.0g Pancreatic digest of casein 7.5g Selected meat peptone 7.5g NaCl 5.0g K 2 HPO 4 4.0g Cornstarch 1.0g KH 2 PO 4 1.0g Supplement solution 10.0mL VCN inhibitor 10.0mL pH 7.2 ± 0.2 at 25°C Source: This medium is available as a premixed powder from BD Di- agnostic Systems . Supplement Solution: Composition per liter: Glucose 100.0g L-Cysteine·HCl 25.9g L-Glutamine 10.0g L-Cystine 1.1g Adenine 1.0g Nicotinamide adenine dinucleotide 0.25g Vitamin B 12 0.1g Thiamine pyrophosphate 0.1g Guanine·HCl 0.03g Fe(NO 3 ) 3 ·6H 2 O 0.02g p-Aminobenzoic acid 0.013g Thiamine·HCl 3.0mg Source: The supplement solution IsoVitaleX ® enrichment is available from BD Diagnostic Systems. This enrichment may be replaced by supplement VX from BD Diagnostic Systems. Preparation of Supplement Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize. VCN Inhibitor: Composition per 10.0mL: Colistin 7.5mg Vancomycin 3.0mg Nystatin 12,500U Preparation of VCN Inhibitor: Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except supplement solution and VCN inhibitor, 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. Cool to 45°–50°C. Aseptically add sterile VCN inhibitor and sterile supplement solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the selective isolation of Neisseria gonorrhoeae and Neis- seria meningitidis from specimens containing mixed flora of bacteria and fungi. Thermincola Medium (DSMZ Medium 1028) Composition per liter: NH 4 Cl 1.0g MgCl 2 ·6H 2 O 0.33g KH 2 PO 4 0.5g KCl 0.33g Na-acetate 0.2g CaCl 2 ·2H 2 O 0.1g Resazurin 0.5mg Vitamin solution 20.0mL Bicarbonate solution 10.0mL Carbonate solution 10.0mL Sulfide solution 10.0mL Yeast extract solution 10.0mL Wolfe's mineral elixir 1.0mL pH 8.0 ± 0.2 at 25°C Sulfide Solution: Composition per 10.0mL: Na 2 S·9H 2 O 1.0g Preparation of Sulfide Solution: Add Na 2 S·9H 2 O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool to room temperature. © 2010 by Taylor and Francis Group, LLC Thermoacetogenium phaeum Medium 1711 Bicarbonate Solution: Composition per 10.0mL: NaHCO 3 0.5g Preparation of Bicarbonate Solution: Add NaHCO 3 to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with gas mixture of 80% N 2 + 20% CO 2 . Filter sterilize. Carbonate Solution: Composition per 10.0mL: Na 2 CO 3 0.5g Preparation of Carbonate Solution: Add Na 2 CO 3 to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with gas mixture of 80% N 2 + 20% CO 2 . Filter sterilize. Yeast Extract Solution: Composition per 10.0mL: Yeast extract 0.2g Preparation of Yeast Extract Solution: Add yeast extract to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 gas. Autoclave for 15 min at 15 psi pressure– 121°C. Wolfe’s Mineral Elixir: Composition per liter: MgSO 4 ·7H 2 O 30.0g NaCl 10.0g MnSO 4 ·2H 2 O 5.0g (NH 4 ) 2 NiSO 4 ·6H 2 O 2.8g CoCl 2 ·6H 2 O 1.8g ZnSO 4 ·7H 2 O 1.8g FeSO 4 ·7H 2 O 1.0g CaCl 2 ·2H 2 O 1.0g KAl(SO 4 ) 2 ·12H 2 O 0.18g CuSO 4 ·5H 2 O 0.1g H 3 BO 3 0.1g Na 2 MoO 4 ·2H 2 O 0.1g Na 2 SeO 4 0.1g Na 2 WO 4 ·2H 2 O 0.1g Preparation of Wolfe’s Mineral Elixir: Adjust pH of 1.0L of distilled/deionized water to 1.0 with dilute H 2 SO 4 . Add components one at a time. Mix thoroughly to dissolve. Vitamin Solution: Composition per liter: Pyridoxine-HCl 10.0mg Thiamine-HCl·2H 2 O 5.0mg Riboflavin 5.0mg Nicotinic acid 5.0mg D-Ca-pantothenate 5.0mg p-Aminobenzoic acid 5.0mg Lipoic acid 5.0mg Biotin 2.0mg Folic acid 2.0mg Vitamin B 12 0.1mg Preparation of Vitamin Solution: Add components to distilled/ deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 80% H 2 + 20% CO 2 . Filter sterilize. Preparation of Medium: Add components, except vitamin solution, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Gently heat and bring to boiling. Boil for 1 min. Cool to room temperature while sparging with 100% N 2 gas. Dispense into culture vessels under an atmostphere of 100% CO (carbon monoxide) . Culture vessels should be filled to approximately 20%. Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature. Aspetical- ly add carbonate, bicarbonate, vitamin, and sulfide solutions. Mix thoroughly. Adjust the pH to 8.0 with sterile, anoxic 1N HCl. Prior to inoculation aseptically add yeast extract solution. Use: For the cultivation of Thermincola carboxydiphila. Thermoacetogenium phaeum Medium (DSMZ Medium 880) Composition per liter: KHCO 3 3.5g NH 4 Cl 1.0g NaCl 0.6g KH 2 PO 4 0.3g MgCl 2 ·6H 2 O 0.1g CaCl 2 ·2H 2 O 0.08g Resazurin 0.5mg Sodium pyruvate solution 50.0mL Vitamin solution 10.0mL Na 2 S·9H 2 O solution 10.0mL Cysteine solution 10.0mL Trace elements solution 1.0mL Selenite-tungstate solution 1.0mL pH 7.0–7.1 at 25°C Sodium Pyruvate Solution: Composition per 50.0mL: Sodium pyruvate 5.0g Preparation of Sodium Pyruvate Solution: Add sodium pyruvate to distilled/deionized water and bring volume to 50.0mL. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Selenite-Tungstate Solution Composition per liter: NaOH 0.5g Na 2 WO 4 ·2H 2 O 4.0mg Na 2 SeO 3 ·5H 2 O 3.0mg Preparation of Selenite-Tungstate Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Na 2 S·9H 2 O Solution: Composition per 10.0mL: Na 2 S·9H 2 O 0.3g Preparation of Na 2 S·9H 2 O Solution: Add Na 2 S·9H 2 O 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. Cysteine Solution: Composition per 10.0mL: L-Cysteine·HCl·H 2 O 0.3g Preparation of Cysteine Solution: Add L-cysteine·HCl·H 2 O 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. Trace Elements Solution SL-10: Composition per liter: FeCl 2 ·4H 2 O 1.5g Na 2 -EDTA 0.5g CoCl 2 ·6H 2 O 190.0mg MnCl 2 ·4H 2 O 100.0mg © 2010 by Taylor and Francis Group, LLC 1712 Thermoacidurans Agar 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 thoroughly. Adjust pH to 6.5. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Vitamin Solution: Composition per liter: Pyridoxine-HCl 10.0mg Thiamine-HCl·2H 2 O 5.0mg Riboflavin 5.0mg Nicotinic acid 5.0mg D-Ca-pantothenate 5.0mg p-Aminobenzoic acid 5.0mg Lipoic acid 5.0mg Biotin 2.0mg Folic acid 2.0mg Vitamin B 12 0.1mg Preparation of Vitamin Solution: Add components to distilled/ deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 80% H 2 + 20% CO 2 . Filter sterilize. Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Add components, except KHCO 3 , sodium pyruvate solution, cysteine solution, and Na 2 S·9H 2 O solution, to distilled/deionized water and bring volume to 930.0mL. Mix thoroughly. Gently heat and bring to boiling. Boil for 10 min. Cool to room temperature while sparging with 80% N 2 + 20% CO 2 . Add 3.5g KHCO 3 . Mix thoroughly while sparging with 80% N 2 + 20% CO 2 gas atmosphere. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add 50.0mL sodium pyruvate solution, 10.0mL cysteine solution, and 10.0mL Na 2 S·9H 2 O solution. Mix thoroughly. Final pH is 7.0– 7.1. Aseptically and anaerobically distribute into sterile tubes or bottles. Use: For the cultivation of Thermacetogenium phaeum. Thermoacidurans Agar Composition per liter: Agar 20.0g Yeast extract 5.0g Proteose peptone 5.0g Glucose 5.0g K 2 HPO 4 4.0g pH 5.0 ± 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. Do not overheat. Pour into sterile Petri dishes or leave in tubes. Use: For the isolation and cultivation of Bacillus thermoacidurans from food products. Thermoacidurans HiVeg Agar Composition per liter: Agar 20.0g Glucose 5.0g Plant peptone No. 3 5.0g Yeast extract 5.0g K 2 HPO 4 4.0g pH 5.0 ± 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 and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 12 psi pressure–118°C. Do not overheat. Pour into sterile Petri dishes or leave in tubes. Use: For the isolation and cultivation of Bacillus thermoacidurans from food products. Thermoactinomyces dichotomicus Medium Composition per liter: Maize, split 50.0g Agar 20.0g Starch 10.0g NaCl 5.0g Peptone 5.0g CaCl 2 0.5g pH 7.2 ± 0.2 at 25°C Preparation of Medium: Add split maize (crushed corn) to 1.0L of boiling water. Steam for 30 min. Filter through Whatman #1 filter pa- per. Add remaining components to maize filtrate. 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 Thermoactinomyces dichotomicus. Thermoactinomyces Medium (DSMZ Medium 978) Composition per liter: Polypeptone™ 30.0g Agar 15.0g Glycerol 2.0g L-Asparagine 1.0g K 2 HPO 4 1.0g Vitamin B solution 10.0mL Trace salts solution 1.0mL pH 7.2 ± 0.2 at 25°C Trace Salts Solution: Composition per 100.0mL: FeSO 4 ·7H 2 O 0.1g MnCl 2 ·4H 2 O 0.1g ZnSO 4 ·7H 2 O 0.1g Preparation of Trace Salts Solution: Add components to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Vitamin B Solution: Composition per 200.0mL: Thiamine-HCl 10.0mg Riboflavin 10.0mg Nicotinate 10.0mg Pyridoxine-HCl 10.0mg © 2010 by Taylor and Francis Group, LLC Thermoanaerobacter ethanolicus Medium 1713 Inositol 10.0mg Calcium pantothenate 10.0mg p-Aminobenzoate 10.0mg D-Biotin 5.0mg Preparation of Vitamin B Solution: Add components to distilled/ deionized water and bring volume to 200.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except vitamin B solution, to distilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature. Aseptically add 100.0mL vitamin B solution. Asep- tically distribute into sterile tubes or flasks. Use: For the cultivation of Thermoactinomyces peptonophilus. Thermoactinomyces Medium Composition per liter: Agar 20.0g Malt extract 10.0g Yeast extract 4.0g Glucose 4.0g pH 7.3 ± 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 Thermoactinomyces sac- chari. Thermoactinomyces Medium (DSMZ Medium 978) Composition per liter: Polypeptone 30.0g Agar 15.0g Glycerol 2.0g L-Asparagine 1.0g KH 2 PO 4 1.0g Vitamin solution 10.0mL Trace elements solution 1.0mL pH 7.2 ± 0.2 at 25°C Vitamin Solution: Composition per 200.0mL: Pyridoxine-HCl 10.0mg Thiamine-HCl·2H 2 O 10.0mg Riboflavin 10.0mg Nicotinic acid 10.0mg D-Ca-pantothenate 10.0mg Inositol 10.0mg p-Aminobenzoic acid 10.0mg Biotin 5.0mg Preparation of Vitamin Solution: Add components to distilled/ deionized water and bring volume to 200.0mL. Mix thoroughly. Filter sterilize. Trace Elements Solution: Composition per 100.0mL: FeSO 4 ·7H 2 O 0.1g MnCl 2 ·4H 2 O 0.1g ZnSO 4 ·7H 2 O 0.1g Preparation of Trace Elements Solution: Add components to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Preparation of Medium: Add components, except vitamin solution, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Adjust the pH to 7.2. Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature. Aspetically add vitamin solution. Mix thoroughly. Aseptically dispense into culture vessels. Use: For the cultivation of Thermoactinomyces spp. Thermoactinopolyspora Medium Composition per liter: Maltose 20.0g Agar 15.0g Papaic digest of soybean meal 15.0g Yeast extract 2.0g pH 7.2 ± 0.2 at 25°C Preparation of Medium: Add components to tap water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Dis- tribute 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 Thermoactinomyces and Thermoactinopolyspora species. Thermoanaerobacter ethanolicus Medium Composition per liter: Glucose 8.0g Na 2 HPO 4 ·12H 2 O 4.2g Yeast extract 2.0g KH 2 PO 4 1.5g NH 4 Cl 0.5g MgCl 2 ·6H 2 O 0.18g Reducing solution 40.0mL Wolfe’s modified mineral solution 5.0mL Resazurin (0.1% solution) 1.0mL Vitamin solution 0.5mL Caution: This medium contains Na 2 S, and H 2 S production will occur, especially upon prolonged boiling. H 2 S is hazardous and preparation of this medium should be done in a chemical fume hood. Reducing Solution: Composition per 200.0mL: Cysteine·HCl·H 2 O 2.5g Na 2 S·9H 2 O 2.5g NaOH (0.2N solution) 200.0mL Preparation of Reducing Solution: Gently heat the NaOH solution and bring to boiling. Gas with 95% N 2 + 5% H 2 . Cool to room temperature. Add the cysteine·HCl·H 2 O and Na 2 S·9H 2 O. Anaerobically distribute into tubes. Cap with rubber stoppers. Autoclave for 15 min at 15 psi pressure–121°C. Vitamin Solution: Composition per 500.0mL: Pyridoxine·HCl 0.1g p-Aminobenzoic acid 0.05g Calcium pantothenate 0.05g Nicotinic acid 0.05g Thioctic acid 0.05g Biotin 0.02g Folic acid 0.02g Riboflavin 5.0mg © 2010 by Taylor and Francis Group, LLC 1714 Thermoanaerobacter subterraneus Medium Thiamine·HCl 5.0mg Vitamin B 12 1.0mg Preparation of Vitamin Solution: Add components to distilled/ deionized water and bring volume to 500.0mL. Mix thoroughly. Store solution in the dark at −10°C. Wolfe’s Modified Mineral Solution: Composition per liter: MgSO 4 ·7H 2 O 3.0g Nitrilotriacetic acid 1.5g NaCl 1.0g MnSO 4 ·H 2 O 0.5g CaCl 2 (anhydrous) 0.1g Co(NO 3 ) 2 ·6H 2 O 0.1g FeSO 4 ·7H 2 O 0.1g ZnSO 4 ·7H 2 O 0.1g AlK(SO 4 ) 2 (anhydrous) 0.01g CuSO 4 ·5H 2 O 0.01g H 3 BO 3 0.01g Na 2 MoO 4 ·2H 2 O 0.01g Na 2 SeO 3 (anhydrous) 1.0mg Preparation of Wolfe’s Modified Mineral Solution: Add nitrilotriacetic acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH to 6.5 with KOH. Add remaining components. Add distilled/deionized water to 1.0L. Preparation of Medium: Add components, except reducing solution, to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling under 95% N 2 + 5% H 2 . Continue boiling until col- or changes from blue to pink. Add the reducing solution. The pink col- or will disappear, indicating that the solution has been reduced. Distribute into tubes or flasks under 95% N 2 + 5% H 2 using anerobic techniques. Cap tubes with rubber stoppers. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation and maintenance of thermophilic anaerobes such as Thermoanaerobacter species and some Clostridium species. Thermoanaerobacter subterraneus Medium (DSMZ Medium 899) Composition per liter: Yeast extract 2.0g MgCl 2 ·6H 2 O 1.0g NH 4 Cl 1.0g NaCl 0.6g Cysteine-HCl·H 2 O 0.5g K 2 HPO 4 0.3g KH 2 PO 4 0.3g KCl 0.2g CaCl 2 ·2H 2 O 0.1g Resazurin 0.5mg D-Glucose solution 30.0mL NaHCO 3 solution 20.0mL Trace mineral solution 10.0mL Na 2 S 2 O 3 solution 10.0mL Na 2 S·9H 2 O solution 10.0mL pH 7.0 ± 0.2 at 25°C Na 2 S·9H 2 O Solution: Composition per 10.0mL: Na 2 S·9H 2 O 0.45g Preparation of Na 2 S·9H 2 O Solution: Add Na 2 S·9H 2 O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool to room temperature. NaHCO 3 Solution: Composition per 20.0mL: NaHCO 3 4.0g Preparation of NaHCO 3 Solution: Add NaHCO 3 to distilled/deionized water and bring volume to 20.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Filter sterilize. Glucose Solution: Composition per 30.0mL: D-Glucose 4.0g Preparation of Glucose Solution: Add D-glucose to distilled/deionized water and bring volume to 30.0mL. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Na 2 S 2 O 3 Solution: Composition per 10.0mL: Na 2 S 2 O 3 ·5H 2 O 2.5g Preparation of Na 2 S 2 O 3 Solution: Add Na 2 S 2 O 3 ·5H 2 O to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Auto- clave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool to room temperature. 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 CaCl 2 ·2H 2 O 0.1g FeSO 4 ·7H 2 O 0.1g NiCl 2 ·6H 2 O 0.025g KAl(SO 4 ) 2 ·12H 2 O 0.02g H 3 BO 3 0.01g Na 2 MoO 4 ·4H 2 O 0.01g CuSO 4 ·5H 2 O 0.01g Na 2 SeO 3 ·5H 2 O 0.3mg Preparation of Trace Elements Solution: Add nitrilotriacetic acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH to 6.5 with KOH. Add remaining components. Add distilled/deionized water to 1.0L. Mix thoroughly. Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Add components, except NaHCO 3 solution, glucose solution, Na 2 S·9H 2 O solution, and Na 2 S 2 O 3 solution, to distilled/deionized water and bring volume to 930.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Distribute into sterile tubes or bottles. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically per 1.0L of medium add 20.0mL NaHCO 3 solution, 30.0mL glucose solution, 10.0mL Na 2 S·9H 2 O solution, and 10.0mL Na 2 S 2 O 3 solution. Mix thoroughly. The final pH should be 7.0. Use: For the cultivation of Thermoanaerobacter subterraneus. Thermoanaerobacter sulfurophilus Medium (DSMZ Medium 827) Composition per 1055.0mL: Sulfur, powdered 10.0g NH 4 Cl 0.33g © 2010 by Taylor and Francis Group, LLC . sterilize. Preparation of Medium: Aseptically combine 500.0mL of sterile solution A, 500.0mL of sterile solution B, 10.0 mL of sterile trace elements solution SL-10, and 5.0 mL of sterile vitamin. add 10.0mL of sterile CNVT inhibitor and 10.0mL of sterile supplement solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the isolation of Neisseria. 0.06mg Preparation of Vitox Supplement: Add components to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: To 740.0mL of cooled sterile