Modified FWM Medium 1205 Solution G 10.0mL Solution B 1.0mL Solution C 1.0mL Solution D 1.0mL pH 7.3 ± 0.2 at 25°C Solution A: Composition per 940.0mL: NaCl 1.0g KCl 0.5g MgCl 2 ·6H 2 O 0.4g NH 4 Cl 0.25g KH 2 PO 4 0.2g CaCl 2 ·2H 2 O 0.15g Resazurin 0.5mg Preparation of Solution A: Prepare under 80% N 2 + 20% CO 2 gas atmosphere. Add components to distilled/deionized water and bring vol- ume to 940.0mL. Mix thoroughly. Adjust pH to 7.2. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution B: 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: Add FeCl 2 ·4H 2 O to 10.0mL of HCl so- lution. Mix thoroughly. Add distilled/deionized water and bring vol- ume to 1.0L. Add remaining components. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution C: Composition per liter: Pyridoxine hydrochloride 300.0mg Thiamine-HCl·2H 2 O 200.0mg Nicotinic acid 200.0mg Vitamin B 12 100.0mg Calcium pantothenate 100.0mg p-Aminobenzoic acid 80.0mg D(+)-Biotin 20.0mg Preparation of Solution C: Add components to distilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 . Mix thorough- ly. Filter sterilize. Solution D: 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 Solution D: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Solution E: Composition per 100.0mL: NaHCO 3 5.0g Preparation of Solution E: Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution F: Composition per 10.0mL: Fructose 2.0g Preparation of Solution F: Add fructose to distilled/deionized wa- ter and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution G: Composition per 10.0mL: Na 2 S·9H 2 O 0.125g Preparation of Solution G: 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 25°C. Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Sequentially add 1.0mL solution B, 1.0mL solution C, 1.0mL solution D, 50.0mL solution E, 10.0mL solu- tion F, and 10.0mL solution G to 940.0mL solution A. Distribute anaer- obically under 80% N 2 + 20% CO 2 into appropriate vessels. The pH should be 7.2–7.4. Use: For the cultivation of Clostridium homopropionicum DSM 5847. Modified FWM Medium (DSMZ Medium 503) Composition per 1013.0mL: Solution A 940.0mL Solution E 50.0mL Solution F 10.0mL Solution G 10.0mL Solution B 1.0mL Solution C 1.0mL Solution D 1.0mL pH 7.3 ± 0.2 at 25°C Solution A: Composition per 940.0mL: NaCl 1.0g KCl 0.5g MgCl 2 ·6H 2 O 0.4g NH 4 Cl 0.25g KH 2 PO 4 0.2g CaCl 2 ·2H 2 O 0.15g Resazurin 0.5mg Preparation of Solution A: Prepare under 80% N 2 + 20% CO 2 gas atmosphere. Add components to distilled/deionized water and bring vol- ume to 940.0mL. Mix thoroughly. Adjust pH to 7.2. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution B: 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 © 2010 by Taylor and Francis Group, LLC 1206 Modified FWM Medium H 3 BO 3 6.0mg CuCl 2 ·2H 2 O 2.0mg HCl (25% solution) 10.0mL Preparation of Solution B: Add FeCl 2 ·4H 2 O to 10.0mL of HCl so- lution. Mix thoroughly. Add distilled/deionized water and bring vol- ume to 1.0L. Add remaining components. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution C: Composition per liter: Pyridoxine hydrochloride 300.0mg Thiamine-HCl·2H 2 O 200.0mg Nicotinic acid 200.0mg Vitamin B 12 100.0mg Calcium pantothenate 100.0mg p-Aminobenzoic acid 80.0mg D(+)-Biotin 20.0mg Preparation of Solution C: Add components to distilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 . Mix thorough- ly. Filter sterilize. Solution D: 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 Solution D: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Solution E: Composition per 100.0mL: NaHCO 3 5.0g Preparation of Solution E: Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution F: Composition per 10.0mL: Na 2 -succinate 2.5g Preparation of Solution F: Add Na 2 -succinate to distilled/deion- ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution G: Composition per 10.0mL: Na 2 S·9H 2 O 0.125g Preparation of Solution G: 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 25°C. Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Sequentially add 1.0mL solution B, 1.0mL solution C, 1.0mL solution D, 50.0mL solution E, 10.0mL so- lution F, and 10.0mL solution G to 940.0mL solution A. Distribute an- aerobically under 80% N 2 + 20% CO 2 into appropriate vessels. The pH should be 7.2–7.4. Use: For the cultivation of unidentified bacterium DSM 5849. Modified FWM Medium (DSMZ Medium 503) Composition per 1013.0mL: Solution A 940.0mL Solution E 50.0mL Solution F 10.0mL Solution G 10.0mL Solution B 1.0mL Solution C 1.0mL Solution D 1.0mL pH 7.3 ± 0.2 at 25°C Solution A: Composition per 940.0mL: NaCl 1.0g KCl 0.5g MgCl 2 ·6H 2 O 0.4g NH 4 Cl 0.25g KH 2 PO 4 0.2g CaCl 2 ·2H 2 O 0.15g Resazurin 0.5mg Preparation of Solution A: Prepare under 80% N 2 + 20% CO 2 gas atmosphere. Add components to distilled/deionized water and bring vol- ume to 940.0mL. Mix thoroughly. Adjust pH to 7.2. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution B: 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: Add FeCl 2 ·4H 2 O to 10.0mL of HCl so- lution. Mix thoroughly. Add distilled/deionized water and bring vol- ume to 1.0L. Add remaining components. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution C: Composition per liter: Pyridoxine hydrochloride 300.0mg Thiamine-HCl·2H 2 O 200.0mg Nicotinic acid 200.0mg Vitamin B 12 100.0mg Calcium pantothenate 100.0mg p-Aminobenzoic acid 80.0mg D(+)-Biotin 20.0mg Preparation of Solution C: Add components to distilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 . Mix thorough- ly. Filter sterilize. Solution D: Composition per liter: NaOH 0.5g Na 2 WO 4 ·2H 2 O 4.0mg Na 2 SeO 3 ·5H 2 O 3.0mg © 2010 by Taylor and Francis Group, LLC Modified FWM Medium 1207 Preparation of Solution D: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Solution E: Composition per 100.0mL: NaHCO 3 5.0g Preparation of Solution E: Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution F: Composition per 10.0mL: Na-(D/L)-3-hydroxybutyrate 1.5g Preparation of Solution F: Add Na-(D/L)-3-hydroxybutyrate to distilled/deionized water and bring volume to 10.0mL. Mix thorough- ly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution G: Composition per 10.0mL: Na 2 S·9H 2 O 0.125g Preparation of Solution G: 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 25°C. Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Sequentially add 1.0mL solution B, 1.0mL solution C, 1.0mL solution D, 50.0mL solution E, 10.0mL so- lution F, and 10.0mL solution G to 940.0mL solution A. Distribute an- aerobically under 80% N 2 + 20% CO 2 into appropriate vessels. The pH should be 7.2–7.4. Use: For the cultivation of Desulfococcus biacutus DSM 5651. Modified FWM Medium (DSMZ Medium 503) Composition per 1013.0mL: Solution A 940.0mL Solution E 50.0mL Solution F 10.0mL Solution G 10.0mL Solution B 1.0mL Solution C 1.0mL Solution D 1.0mL pH 7.3 ± 0.2 at 25°C Solution A: Composition per 940.0mL: NaCl 1.0g KCl 0.5g MgCl 2 ·6H 2 O 0.4g NH 4 Cl 0.25g KH 2 PO 4 0.2g CaCl 2 ·2H 2 O 0.15g Resazurin 0.5mg Preparation of Solution A: Prepare under 80% N 2 + 20% CO 2 gas atmosphere. Add components to distilled/deionized water and bring vol- ume to 940.0mL. Mix thoroughly. Adjust pH to 7.2. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution B: 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: Add FeCl 2 ·4H 2 O to 10.0mL of HCl so- lution. Mix thoroughly. Add distilled/deionized water and bring vol- ume to 1.0L. Add remaining components. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution C: Composition per liter: Pyridoxine hydrochloride 300.0mg Thiamine-HCl·2H 2 O 200.0mg Nicotinic acid 200.0mg Vitamin B 12 100.0mg Calcium pantothenate 100.0mg p-Aminobenzoic acid 80.0mg D(+)-Biotin 20.0mg Preparation of Solution C: Add components to distilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 . Mix thorough- ly. Filter sterilize. Solution D: 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 Solution D: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Solution E: Composition per 100.0mL: NaHCO 3 5.0g Preparation of Solution E: Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution F: Composition per 10.0mL: Xylan 2.0g Preparation of Solution F: Add xylan to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution G: Composition per 10.0mL: Na 2 S·9H 2 O 0.125g Preparation of Solution G: 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 25°C. © 2010 by Taylor and Francis Group, LLC 1208 Modified FWM Medium Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Sequentially add 1.0mL solution B, 1.0mL solution C, 1.0mL solution D, 50.0mL solution E, 10.0mL so- lution F, and 10.0mL solution G to 940.0mL solution A. Distribute an- aerobically under 80% N 2 + 20% CO 2 into appropriate vessels. The pH should be 7.2–7.4. Use: For the cultivation of Cytophaga xylanolytica DSM 6779. Modified FWM Medium (DSMZ Medium 503) Composition per 1013.0mL: Solution A 940.0mL Solution E 50.0mL Solution F 10.0mL Solution G 10.0mL Solution B 1.0mL Solution C 1.0mL Solution D 1.0mL pH 7.3 ± 0.2 at 25°C Solution A: Composition per 940.0mL: NaCl 1.0g KCl 0.5g MgCl 2 ·6H 2 O 0.4g NH 4 Cl 0.25g KH 2 PO 4 0.2g CaCl 2 ·2H 2 O 0.15g Resazurin 0.5mg Preparation of Solution A: Prepare under 80% N 2 + 20% CO 2 gas atmosphere. Add components to distilled/deionized water and bring vol- ume to 940.0mL. Mix thoroughly. Adjust pH to 7.2. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution B: 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: Add FeCl 2 ·4H 2 O to 10.0mL of HCl so- lution. Mix thoroughly. Add distilled/deionized water and bring vol- ume to 1.0L. Add remaining components. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution C: Composition per liter: Pyridoxine hydrochloride 300.0mg Thiamine-HCl·2H 2 O 200.0mg Nicotinic acid 200.0mg Vitamin B 12 100.0mg Calcium pantothenate 100.0mg p-Aminobenzoic acid 80.0mg D(+)-Biotin 20.0mg Preparation of Solution C: Add components to distilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 . Mix thorough- ly. Filter sterilize. Solution D: 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 Solution D: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Solution E: Composition per 100.0mL: NaHCO 3 5.0g Preparation of Solution E: Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution F: Composition per 10.0mL: Xylose 2.0g Preparation of Solution F: Add xylose to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution G: Composition per 10.0mL: Na 2 S·9H 2 O 0.125g Preparation of Solution G: 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 25°C. Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Sequentially add 1.0mL solution B, 1.0mL solution C, 1.0mL solution D, 50.0mL solution E, 10.0mL so- lution F, and 10.0mL solution G to 940.0mL solution A. Distribute an- aerobically under 80% N 2 + 20% CO 2 into appropriate vessels. The pH should be 7.2–7.4. Use: For the cultivation of Cytophaga xylanolytica DSM 6779. Modified FWM Medium (DSMZ Medium 503) Composition per 1013.0mL: Solution A 940.0mL Solution E 50.0mL Solution F 10.0mL Solution G 10.0mL Solution B 1.0mL Solution C 1.0mL Solution D 1.0mL pH 7.3 ± 0.2 at 25°C Solution A: Composition per 940.0mL: NaCl 1.0g KCl 0.5g MgCl 2 ·6H 2 O 0.4g NH 4 Cl 0.25g KH 2 PO 4 0.2g © 2010 by Taylor and Francis Group, LLC Modified FWM Medium 1209 CaCl 2 ·2H 2 O 0.15g Resazurin 0.5mg Preparation of Solution A: Prepare under 80% N 2 + 20% CO 2 gas atmosphere. Add components to distilled/deionized water and bring vol- ume to 940.0mL. Mix thoroughly. Adjust pH to 7.2. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution B: 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: Add FeCl 2 ·4H 2 O to 10.0mL of HCl so- lution. Mix thoroughly. Add distilled/deionized water and bring vol- ume to 1.0L. Add remaining components. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution C: Composition per liter: Pyridoxine hydrochloride 300.0mg Thiamine-HCl·2H 2 O 200.0mg Nicotinic acid 200.0mg Vitamin B 12 100.0mg Calcium pantothenate 100.0mg p-Aminobenzoic acid 80.0mg D(+)-Biotin 20.0mg Preparation of Solution C: Add components to distilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 . Mix thoroughly. Filter sterilize. Solution D: 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 Solution D: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Solution E: Composition per 100.0mL: NaHCO 3 5.0g Preparation of Solution E: Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution F: Composition per 10.0mL: Pivalic acid 1.0g NaNO 3 0.85g Preparation of Solution F: Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution G: Composition per 10.0mL: Na 2 S·9H 2 O 0.125g Preparation of Solution G: 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 25°C. Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Sequentially add 1.0mL solution B, 1.0mL solution C, 1.0mL solution D, 50.0mL solution E, 10.0mL so- lution F, and 10.0mL solution G to 940.0mL solution A. Distribute an- aerobically under 80% N 2 + 20% CO 2 into appropriate vessels. The pH should be 7.2–7.4. Use: For the cultivation of Thavera pivalivorans DSM 14691. Modified FWM Medium (DSMZ Medium 503) Composition per 1013.0mL: Solution A 940.0mL Solution E 50.0mL Solution F 10.0mL Solution G 10.0mL Solution B 1.0mL Solution C 1.0mL Solution D 1.0mL pH 7.3 ± 0.2 at 25°C Solution A: Composition per 940.0mL: NaCl 1.0g KCl 0.5g MgCl 2 ·6H 2 O 0.4g NH 4 Cl 0.25g KH 2 PO 4 0.2g CaCl 2 ·2H 2 O 0.15g Resazurin 0.5mg Preparation of Solution A: Prepare under 80% N 2 + 20% CO 2 gas atmosphere. Add components to distilled/deionized water and bring vol- ume to 940.0mL. Mix thoroughly. Adjust pH to 7.2. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution B: 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: Add FeCl 2 ·4H 2 O to 10.0mL of HCl so- lution. Mix thoroughly. Add distilled/deionized water and bring vol- ume to 1.0L. Add remaining components. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. © 2010 by Taylor and Francis Group, LLC 1210 Modified FWM Medium Solution C: Composition per liter: Pyridoxine hydrochloride 300.0mg Thiamine-HCl·2H 2 O 200.0mg Nicotinic acid 200.0mg Vitamin B 12 100.0mg Calcium pantothenate 100.0mg p-Aminobenzoic acid 80.0mg D(+)-Biotin 20.0mg Preparation of Solution C: Add components to distilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 . Mix thorough- ly. Filter sterilize. Solution D: 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 Solution D: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Solution E: Composition per 100.0mL: NaHCO 3 5.0g Preparation of Solution E: Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution F: Composition per 10.0mL: Glucose 5.0g Yeast extract 2.0g Preparation of Solution F: Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution G: Composition per 10.0mL: Na 2 S·9H 2 O 0.125g Preparation of Solution G: 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 25°C. Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Sequentially add 1.0mL solution B, 1.0mL solution C, 1.0mL solution D, 50.0mL solution E, 10.0mL so- lution F, and 10.0mL solution G to 940.0mL solution A. Distribute an- aerobically under 80% N 2 + 20% CO 2 into appropriate vessels. The pH should be 7.2–7.4. Use: For the cultivation of Opitutus sp. DSM 14424. Modified FWM Medium (DSMZ Medium 503) Composition per 1013.0mL: Solution A 940.0mL Solution E 50.0mL Solution F 10.0mL Solution G 10.0mL Solution B 1.0mL Solution C 1.0mL Solution D 1.0mL pH 7.3 ± 0.2 at 25°C Solution A: Composition per 940.0mL: NaCl 1.0g KCl 0.5g MgCl 2 ·6H 2 O 0.4g NH 4 Cl 0.25g KH 2 PO 4 0.2g CaCl 2 ·2H 2 O 0.15g Resazurin 0.5mg Preparation of Solution A: Prepare under 80% N 2 + 20% CO 2 gas atmosphere. Add components to distilled/deionized water and bring vol- ume to 940.0mL. Mix thoroughly. Adjust pH to 7.2. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution B: 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: Add FeCl 2 ·4H 2 O to 10.0mL of HCl so- lution. Mix thoroughly. Add distilled/deionized water and bring vol- ume to 1.0L. Add remaining components. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution C: Composition per liter: Pyridoxine hydrochloride 300.0mg Thiamine-HCl·2H 2 O 200.0mg Nicotinic acid 200.0mg Vitamin B 12 100.0mg Calcium pantothenate 100.0mg p-Aminobenzoic acid 80.0mg D(+)-Biotin 20.0mg Preparation of Solution C: Add components to distilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 . Mix thorough- ly. Filter sterilize. Solution D: 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 Solution D: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Solution E: Composition per 100.0mL: NaHCO 3 5.0g Preparation of Solution E: Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with © 2010 by Taylor and Francis Group, LLC Modified FWM Medium 1211 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution F: Composition per 10.0mL: Na 2 -fumarate 3.2g Na-acetate 0.8g Preparation of Solution F: Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution G: Composition per 10.0mL: Na 2 S·9H 2 O 0.125g Preparation of Solution G: 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 25°C. Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Sequentially add 1.0mL solution B, 1.0mL solution C, 1.0mL solution D, 50.0mL solution E, 10.0mL so- lution F, and 10.0mL solution G to 940.0mL solution A. Distribute an- aerobically under 80% N 2 + 20% CO 2 into appropriate vessels. The pH should be 7.2–7.4. Use: For the cultivation of Geovibrio thiophilus DSM 11263. Modified FWM Medium (DSMZ Medium 503) Composition per 1013.0mL: Solution A 940.0mL Solution E 50.0mL Solution F 10.0mL Solution G 10.0mL Solution B 1.0mL Solution C 1.0mL Solution D 1.0mL pH 7.3 ± 0.2 at 25°C Solution A: Composition per 940.0mL: NaCl 1.0g KCl 0.5g MgCl 2 ·6H 2 O 0.4g NH 4 Cl 0.25g KH 2 PO 4 0.2g CaCl 2 ·2H 2 O 0.15g Resazurin 0.5mg Preparation of Solution A: Prepare under 80% N 2 + 20% CO 2 gas atmosphere. Add components to distilled/deionized water and bring vol- ume to 940.0mL. Mix thoroughly. Adjust pH to 7.2. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution B: 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: Add FeCl 2 ·4H 2 O to 10.0mL of HCl so- lution. Mix thoroughly. Add distilled/deionized water and bring vol- ume to 1.0L. Add remaining components. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution C: Composition per liter: Pyridoxine hydrochloride 300.0mg Thiamine-HCl·2H 2 O 200.0mg Nicotinic acid 200.0mg Vitamin B 12 100.0mg Calcium pantothenate 100.0mg p-Aminobenzoic acid 80.0mg D(+)-Biotin 20.0mg Preparation of Solution C: Add components to distilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 . Mix thorough- ly. Filter sterilize. Solution D: 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 Solution D: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Solution E: Composition per 100.0mL: NaHCO 3 5.0g Preparation of Solution E: Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution F: Composition per 10.0mL: Na-glycolate 2.0g Yeast extract 1.0g Preparation of Solution F: Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution G: Composition per 10.0mL: Na 2 S·9H 2 O 0.125g Preparation of Solution G: 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 25°C. Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Sequentially add 1.0mL solution B, 1.0mL solution C, 1.0mL solution D, 50.0mL solution E, 10.0mL so- lution F, and 10.0mL solution G to 940.0mL solution A. Distribute an- aerobically under 80% N 2 + 20% CO 2 into appropriate vessels. The pH should be 7.2–7.4. Use: For the cultivation of Clostridium sp. DSM 11261. © 2010 by Taylor and Francis Group, LLC 1212 Modified FWM Medium Modified FWM Medium (DSMZ Medium 503) Composition per 1013.0mL: Solution A 940.0mL Solution E 50.0mL Solution F 10.0mL Solution G 10.0mL Solution B 1.0mL Solution C 1.0mL Solution D 1.0mL pH 7.3 ± 0.2 at 25°C Solution A: Composition per 940.0mL: NaCl 1.0g KCl 0.5g MgCl 2 ·6H 2 O 0.4g NH 4 Cl 0.25g KH 2 PO 4 0.2g CaCl 2 ·2H 2 O 0.15g Resazurin 0.5mg Preparation of Solution A: Prepare under 80% N 2 + 20% CO 2 gas atmosphere. Add components to distilled/deionized water and bring vol- ume to 940.0mL. Mix thoroughly. Adjust pH to 7.2. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution B: 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: Add FeCl 2 ·4H 2 O to 10.0mL of HCl so- lution. Mix thoroughly. Add distilled/deionized water and bring vol- ume to 1.0L. Add remaining components. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution C: Composition per liter: Pyridoxine hydrochloride 300.0mg Thiamine-HCl·2H 2 O 200.0mg Nicotinic acid 200.0mg Vitamin B 12 100.0mg Calcium pantothenate 100.0mg p-Aminobenzoic acid 80.0mg D(+)-Biotin 20.0mg Preparation of Solution C: Add components to distilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 . Mix thorough- ly. Filter sterilize. Solution D: 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 Solution D: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Solution E: Composition per 100.0mL: NaHCO 3 5.0g Preparation of Solution E: Add NaHCO 3 to distilled/deionized wa- ter and bring volume to 100.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution F: Composition per 10.0mL: Na-3-hydroxybutyrate 3.0g Yeast extract 1.0g Preparation of Solution F: Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution G: Composition per 10.0mL: Na 2 S·9H 2 O 0.125g Preparation of Solution G: 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 25°C. Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Sequentially add 1.0mL solution B, 1.0mL solution C, 1.0mL solution D, 50.0mL solution E, 10.0mL so- lution F, and 10.0mL solution G to 940.0mL solution A. Distribute an- aerobically under 80% N 2 + 20% CO 2 into appropriate vessels. The pH should be 7.2–7.4. Use: For the cultivation of unclassified bacterium DSM 11262. Modified FWM Medium (DSMZ Medium 503) Composition per 1023.0mL: Solution A 940.0mL Solution E 50.0mL Solution F 10.0mL Solution G 20.0mL Solution B 1.0mL Solution C 1.0mL Solution D 1.0mL pH 7.3 ± 0.2 at 25°C Solution A: Composition per 940.0mL: Na 2 SO 4 2.8 g NaCl 1.0g KCl 0.5g MgCl 2 ·6H 2 O 0.4g NH 4 Cl 0.25g KH 2 PO 4 0.2g CaCl 2 ·2H 2 O 0.15g Resazurin 0.5mg Preparation of Solution A: Prepare under 80% N 2 + 20% CO 2 gas atmosphere. Add components to distilled/deionized water and bring vol- ume to 940.0mL. Mix thoroughly. Adjust pH to 7.2. Sparge with 80% N 2 © 2010 by Taylor and Francis Group, LLC Modified FWM Medium 1213 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution B: 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: Add FeCl 2 ·4H 2 O to 10.0mL of HCl so- lution. Mix thoroughly. Add distilled/deionized water and bring vol- ume to 1.0L. Add remaining components. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution C: Composition per liter: Pyridoxine hydrochloride 300.0mg Thiamine-HCl·2H 2 O 200.0mg Nicotinic acid 200.0mg Vitamin B 12 100.0mg Calcium pantothenate 100.0mg p-Aminobenzoic acid 80.0mg D(+)-Biotin 20.0mg Preparation of Solution C: Add components to distilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 . Mix thorough- ly. Filter sterilize. Solution D: 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 Solution D: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Solution E: Composition per 100.0mL: NaHCO 3 5.0g Preparation of Solution E: Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution F: Composition per 10.0mL: Syringic acid 0.6 g Preparation of Solution F: Add syringic acid to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Adjust pH to 7.0. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pres- sure–121°C. Cool to 25°C. Solution G: Composition per 20.0mL: Na 2 S·9H 2 O 0.25g Preparation of Solution G: Add Na 2 S·9H 2 O to distilled/deionized water and bring volume to 20.0mL. Mix thoroughly. Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool to 25°C. Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Sequentially add 1.0mL solution B, 1.0mL solution C, 1.0mL solution D, 50.0mL solution E, 10.0mL so- lution F, and 20.0mL solution G to 940.0mL solution A. Distribute an- aerobically under 80% N 2 + 20% CO 2 into appropriate vessels. The pH should be 7.2–7.4. Prior to inoculation the completed medium should equilibrate overnight. Use: For the cultivation of Parasporobacterium paucivorans DSM 15970. Modified FWM Medium (DSMZ Medium 503) Composition per 1023.0mL: Solution A 940.0mL Solution E 50.0mL Solution F 10.0mL Solution G 10.0mL Solution H 10.0mL Solution B 1.0mL Solution C 1.0mL Solution D 1.0mL pH 6.5–7.0 at 25°C Solution A: Composition per 940.0mL: NaCl 1.0g KCl 0.5g MgCl 2 ·6H 2 O 0.4g NH 4 Cl 0.25g KH 2 PO 4 0.2g CaCl 2 ·2H 2 O 0.15g Resazurin 0.5mg Preparation of Solution A: Prepare under 80% N 2 + 20% CO 2 gas atmosphere. Add components to distilled/deionized water and bring vol- ume to 940.0mL. Mix thoroughly. Adjust pH to 7.2. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution B: 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: Add FeCl 2 ·4H 2 O to 10.0mL of HCl so- lution. Mix thoroughly. Add distilled/deionized water and bring vol- ume to 1.0L. Add remaining components. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution C: Composition per liter: Pyridoxine hydrochloride 300.0mg Thiamine-HCl·2H 2 O 200.0mg © 2010 by Taylor and Francis Group, LLC 1214 Modified Gorodkowa Agar Nicotinic acid 200.0mg Vitamin B 12 100.0mg Calcium pantothenate 100.0mg p-Aminobenzoic acid 80.0mg D(+)-Biotin 20.0mg Preparation of Solution C: Add components to distilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 . Mix thorough- ly. Filter sterilize. Solution D: 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 Solution D: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Solution E: Composition per 100.0mL: NaHCO 3 5.0g Preparation of Solution E: Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution F: Composition per 10.0mL: Trimethylamine hydrochloride 2.0g Methanol 0.6g Preparation of Solution F: Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution G: Composition per 10.0mL: Na 2 S·9H 2 O 0.125g Preparation of Solution G: 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 25°C. Solution H: Composition per 10.0mL: FeSO 4 ·6H 2 O 0.02g Preparation of Solution H: Add FeSO 4 ·6H 2 O to distilled/deion- ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Sequentially add 1.0mL solution B, 1.0mL solution C, 1.0mL solution D, 50.0mL solution E, 10.0mL so- lution F, and 10.0mL solution G to 940.0mL solution A. Distribute an- aerobically under 80% N 2 + 20% CO 2 into appropriate vessels. The pH should be 6.5–7.0. After inoculation aseptically inject 0.1mL of so- lution H for each 10.0mL of medium. Use: For the cultivation of Methanomethylovorans hollandica DSM 15978. Modified Gorodkowa Agar Composition per liter: Agar 20.0g Peptic digest of animal tissue 10.0g NaCl 5.0g Glucose 1.0g pH 6.8 ± 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 with frequent agitation. 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 promoting sporulation of yeasts. Modified ISP5 Medium (DSMZ Medium 993b) Composition per liter: KCl 100.0 Agar 20.0g Glycerol 10.0g Yeast extract 5.0g L-asparagine, anhydrous 1.0g K 2 HPO 4 , anhydrous 1.0g Trace elements solution 1.0mL pH 7.2 ± 0.2 at 25°C 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 thorough- ly. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.2. 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. Pour into Petri dishes or leave in tubes. Use: For the cultivation of Nesterenkonia xinjiangensis and Micro- bacterium halotolerans. Modified ISP5 Medium with Sodium Chloride (DSMZ Medium 993c) Composition per liter: NaCl 100.0g Agar 20.0g Glycerol 10.0g L-asparagine, anhydrous 1.0g K 2 HPO 4 , anhydrous 1.0g Trace elements solution 1.0mL pH 7.2 ± 0.2 at 25°C 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 © 2010 by Taylor and Francis Group, LLC . 0.1mL of so- lution H for each 10.0mL of medium. Use: For the cultivation of Methanomethylovorans hollandica DSM 15978. Modified Gorodkowa Agar Composition per liter: Agar 20.0g Peptic digest of. Medium H 3 BO 3 6.0mg CuCl 2 ·2H 2 O 2.0mg HCl (25% solution) 10.0mL Preparation of Solution B: Add FeCl 2 ·4H 2 O to 10.0mL of HCl so- lution. Mix thoroughly. Add distilled/deionized water and bring. 24.0mg H 3 BO 3 6.0mg CuCl 2 ·2H 2 O 2.0mg HCl (25% solution) 10.0mL Preparation of Solution B: Add FeCl 2 ·4H 2 O to 10.0mL of HCl so- lution. Mix thoroughly. Add distilled/deionized water and bring