Handbook of Microbiological Media, Fourth Edition part 59 doc

Desulfuromonas acetexigenes Medium 575 Preparation of NaHCO 3 Solution: Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Filter sterilize. Na-pyruvate Solution: Composition per 10.0mL: Na-pyruvate 0.6g Preparation of Na-pyruvate Solution: Add Na-pyruvate to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Seven Vitamin Solution: 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 Seven Vitamin Solution: Add components to distilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 . Mix thoroughly. Filter sterilize. 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 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: Sulfur is sterilized by steaming for 3 hr on each of 3 successive days. Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Add components, except sulfur, NaHCO 3 solution, Na-pyruvate solution, Na 2 S·9H 2 O solution, seven vitamin solution, and trace elements solution SL-10, to distilled/deionized water and bring volume to 945.0mL. 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. Aseptically and anaerobically add 5.0g sterile sulfur, 33.0mL NaHCO 3 solution, 10.0mL Na-pyruvate solution, 10.0mL Na 2 S·9H 2 O solution, 1.0mL seven vitamin solution, and 1.0mL trace elements solution SL-10. Mix thoroughly. Aseptically and anaerobically distribute into sterile tubes or bottles. Use: For the cultivation of Desulfuromonas thiophila and Desulfu- romonas acetexigens. Desulfuromonas acetexigenes Medium (DSMZ Medium 647) Composition per liter: NaCl 20.0g Sulfur, powdered 5.0g MgCl 2 ·6H 2 O 2.0g KH 2 PO 4 1.0g MgSO 4 ·7H 2 O 1.0g NH 4 Cl 0.5g CaCl 2 ·2H 2 O 0.1g NaHCO 3 solution 50.0mL Na-pyruvate solution 10.0mL Na 2 S·9H 2 O solution 10.0mL Seven vitamin solution 1.0mL Trace elements solution SL-10 1.0mL pH 7.2 ± 0.2 at 25°C 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 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 100.0mL: NaHCO 3 5.0g Preparation of NaHCO 3 Solution: Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Filter sterilize. Na-pyruvate Solution: Composition per 10.0mL: Na-pyruvate 0.6g Preparation of Na-pyruvate Solution: Add Na-pyruvate to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Seven Vitamin Solution: 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 Seven Vitamin Solution: Add components to distilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 . Mix thoroughly. Filter sterilize. 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 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. © 2010 by Taylor and Francis Group, LLC 576 Desulfuromonas acetoxidans Medium Preparation of Medium: Sulfur is sterilized by steaming for 3 hr on each of 3 successive days. Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Add components, except sulfur, NaHCO 3 solution, Na-pyruvate solution, Na 2 S·9H 2 O solution, seven vitamin solution, and trace elements solution SL-10, to distilled/deionized water and bring volume to 928.0mL. 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. Aseptically and anaerobically add 5.0g sterile sulfur, 50.0mL NaHCO 3 solution, 10.0mL Na-pyruvate solution, 10.0mL Na 2 S·9H 2 O solution, 1.0mL seven vitamin solution, and 1.0mL trace elements solution SL-10. Mix thoroughly. Aseptically and anaerobically distribute into sterile tubes or bottles. Use: For the cultivation of Desulfuromonas acetexigenes DSM8988. Desulfuromonas acetoxidans Medium Composition per 1001.0mL: Fumaric acid 1.5g KH 2 PO 4 1.0g NH 4 Cl 0.5g Sodium acetate 0.5g MgSO 4 ·7H 2 O 0.4g CaCl 2 ·2H 2 O 0.1g Resazurin 0.5mg NaHCO 3 solution 40.0mL Trace elements solution SL-4 10.0mL Na 2 S·9H 2 O solution 6.0mL Vitamin solution 5.0mL pH 7.5 ± 0.2 at 25°C NaHCO 3 Solution: Composition per 40.0mL: NaHCO 3 2.0g Preparation of NaHCO 3 Solution: Add NaHCO 3 to distilled/deionized water and bring volume to 40.0mL. Mix thoroughly. Gas under 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Trace Elements Solution SL-4: Composition per liter: EDTA 0.5g FeSO 4 ·7H 2 O 0.2g Trace elements solution SL-6 100.0mL Trace Elements Solution SL-6: Composition per liter: MnCl 2 ·4H 2 O 0.5g H 3 BO 3 0.3g CoCl 2 ·6H 2 O 0.2g ZnSO 4 ·7H 2 O 0.1g Na 2 MoO 4 ·2H 2 O 0.03g NiCl 2 ·6H 2 O 0.02g CuCl 2 ·2H 2 O 0.01g Preparation of Trace Elements Solution SL-6: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gas under 100% N 2 . Autoclave for 15 min at 15 psi pressure– 121°C. Preparation of Trace Elements Solution SL-4: Add components to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Na 2 S·9H 2 O Solution: Composition per 6.0mL: 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. Gas under 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Vitamin Solution: Composition per liter: Pyridoxine·HCl 62.5g Nicotinic acid 25.0mg p-Aminobenzoic acid 12.5mg Thiamine·HCl 12.5mg Calcium DL-pantothenate 6.5mg Biotin 2.5mg Preparation of Vitamin Solution: Add components to distilled/ deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.5. Gas under 100% N 2 . Filter sterilize. Preparation of Medium: Add components, except NaHCO 3 solution, Na 2 S·9H 2 O solution, and vitamin solution, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 5.0. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add 40.0mL of sterile NaHCO 3 solution, 6.0mL of sterile Na 2 S·9H 2 O solution, and 5.0mL of sterile vitamin solution. Mix thoroughly. Aseptically and anaerobically distribute into sterile screw-capped bottles or tubes. Fill completely, leaving only a small gas bubble. Use: For the cultivation and maintenance of Desulfuromonas acetoxidans. Desulfuromonas Medium Composition per 1051.0mL: Elemental sulfur slurry 10.0g Solution 1 1.0L Solution 3 40.0mL Solution 4 6.0mL Solution 5 5.0mL Solution 2 1.0mL pH 7.2 ± 0.2 at 25°C Solution 1: Composition per liter: NaCl 20.0g MgCl 2 ·6H 2 O 3.0g KH 2 PO 4 1.0g NH 4 Cl 0.3g CaCl 2 ·2H 2 O 0.1g HCl (2N solution) 4.0mL Preparation of Solution 1: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution 2: Composition per liter: Disodium EDTA 5.2g CoCl 2 ·6H 2 O 1.9g FeCl 2 ·4H 2 O 1.5g MnCl 2 ·4H 2 O 1.0g ZnCl 2 0.7g H 3 BO 3 0.62g Na 2 MoO 4 ·2H 2 O 0.36g NiCl 2 ·6H 2 O 0.24g CuCl 2 ·2H 2 O 0.17g pH 6.5 ± 0.2 at 25°C © 2010 by Taylor and Francis Group, LLC Desulfuromonas Medium 577 Preparation of Solution 2: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 6.5. Au- toclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution 3: Composition per 100.0mL: NaHCO 3 10.0g Preparation of Solution 3: Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution 4: Composition per 100.0mL: Na 2 S·9H 2 O 5.0g Preparation of Solution 4: Add Na 2 S·9H 2 O to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution 5: Composition per 200.0mL: Pyridoxamine·HCl 0.01g Nicotinic acid 4.0mg p-Aminobenzoic acid 2.0mg Thiamine 2.0mg Cyanocobalamin 1.0mg Pantothenic acid 1.0mg Biotin 0.5mg Preparation of Solution 5: Add components to distilled/deionized water and bring volume to 200.0mL. Mix thoroughly. Filter sterilize. Elemental Sulfur Slurry: Composition per 10.0g: Sulfur flowers 10.0g Preparation of Elemental Sulfur Slurry: Add highly purified sulfur flowers to a mortar and grind to a fine powder. Add sufficient distilled/deionized water to produce a slurry. Distribute into 100.0mL screw-capped bottles in 20.0mL volumes. Autoclave for 30 min at 10 psi pressure–115°C. Decant supernatant solution. Reserve sulfur slurry. Preparation of Medium: To 1.0L of cooled, sterile solution 1, aseptically add 1.0mL of sterile solution 2, 40.0mL of sterile solution 3, 6.0mL of sterile solution 4, and 5.0mL of sterile solution 5. Mix thoroughly. Adjust pH to 7.2. Aseptically distribute into sterile 50.0mL screw-capped bottles. Fill bottles completely with medium except for a pea-sized air bubble. Aseptically add a pea-sized piece of sulfur slurry to each 50.0mL of medium. Use: For the isolation and cultivation of marine Desulfuromonas species. Desulfuromonas Medium Composition per 1031.0mL: Elemental sulfur slurry 10.0g Solution 1 1.0L Solution 3 20.0mL Solution 4 6.0mL Solution 5 5.0mL Solution 2 1.0mL pH 7.2 ± 0.2 at 25°C Solution 1: Composition per liter: KH 2 PO 4 1.0g MgCl 2 ·6H 2 O 0.4g NH 4 Cl 0.3g CaCl 2 ·2H 2 O 0.1g HCl (2N solution) 4.0mL Preparation of Solution 1: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution 2: Composition per liter: Disodium EDTA 5.2g CoCl 2 ·6H 2 O 1.9g FeCl 2 ·4H 2 O 1.5g MnCl 2 ·4H 2 O 1.0g ZnCl 2 0.7g H 3 BO 3 0.62g Na 2 MoO 4 ·2H 2 O 0.36g NiCl 2 ·6H 2 O 0.24g CuCl 2 ·2H 2 O 0.17g Preparation of Solution 2: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 6.5. Au- toclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution 3: Composition per 100.0mL: NaHCO 3 10.0g Preparation of Solution 3: Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution 4: Composition per 100.0mL: Na 2 S·9H 2 O 5.0g Preparation of Solution 4: Add Na 2 S·9H 2 O to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution 5: Composition per 200.0mL: Pyridoxamine·HCl 0.01g Nicotinic acid 4.0mg p-Aminobenzoic acid 2.0mg Thiamine 2.0mg Cyanocobalamin 1.0mg Pantothenic acid 1.0mg Biotin 0.5mg Preparation of Solution 5: Add components to distilled/deionized water and bring volume to 200.0mL. Mix thoroughly. Filter sterilize. Elemental Sulfur Slurry: Composition per 10.0g: Sulfur flowers 10.0g Preparation of Elemental Sulfur Slurry: Add highly purified sulfur flowers to a mortar and grind to a fine powder. Add sufficient distilled/deionized water to produce a slurry. Distribute into 100.0mL screw-capped bottles in 20.0mL volumes. Autoclave for 30 min at 10 psi pressure–115°C. Decant supernatant solution. Reserve sulfur slurry. Preparation of Medium: To 1.0L of cooled, sterile solution 1, aseptically add 1.0mL of sterile solution 2, 40.0mL of sterile solution 3, 6.0mL of sterile solution 4, and 5.0mL of sterile solution 5. Mix thoroughly. Adjust pH to 7.2. Aseptically distribute into sterile 50.0mL screw-capped bottles. Fill bottles completely with medium except for © 2010 by Taylor and Francis Group, LLC 578 Desulfuromonas Medium a pea-sized air bubble. Aseptically add a pea-sized piece of sulfur slurry to each 50.0mL of medium. Use: For the isolation and cultivation of freshwater Desulfuromonas species. Desulfuromonas Medium Composition per 1003.0mL: Solution A 950.0mL Solution C 40.0mL Solution D 10.0mL Solution B 3.0mL pH 7.2 ± 0.2 at 25°C Solution A: Composition per 950.0mL: NaCl 20.0g Disodium DL-malate 2.66g MgCl 2 ·6H 2 O 2.0g Na 2 SO 4 1.77g MgSO 4 ·7H 2 O 1.0g KH 2 PO 4 0.5g Yeast extract 0.5g NH 4 Cl 0.3g CaCl 2 ·2H 2 O 0.1g Resazurin 1.0mg Trace elements solution SL-4 10.0mL Trace Elements Solution SL-4: Composition per liter: EDTA 0.5g FeSO 4 ·7H 2 O 0.2g Trace elements solution SL-6 100.0mL Trace Elements Solution SL-6: Composition per liter: MnCl 2 ·4H 2 O 0.5g H 3 BO 3 0.3g CoCl 2 ·6H 2 O 0.2g ZnSO 4 ·7H 2 O 0.1g Na 2 MoO 4 ·2H 2 O 0.03g NiCl 2 ·6H 2 O 0.02g CuCl 2 ·2H 2 O 0.01g Preparation of Trace Elements Solution SL-6: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gas under 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Trace Elements Solution SL-4: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Preparation of Solution A: Add components to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Adjust pH to 6.0 with 2N NaOH. Gently heat and bring to boiling. Continue boiling for 3–4 min. Allow to cool to room temperature while gassing under 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Solution B: Composition per 3.0mL: Ethanol 0.3mL Preparation of Solution B: Add ethanol to distilled/deionized water and bring volume to 3.0mL. Sparge under 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Solution C: Composition per 40.0mL: NaHCO 3 1.85g Preparation of Solution C: Add NaHCO 3 to distilled/deionized water and bring volume to 40.0mL. Mix thoroughly. Gas under 80% N 2 + 20% CO 2 . Filter sterilize. Solution D: Composition per 10.0mL: Na 2 S·9H 2 O 0.3g Preparation of Solution D: Add Na 2 S·9H 2 O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Gas under 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: To 950.0mL of sterile solution A, aseptically and anaerobically add 3.0mL of sterile solution B, 40.0mL of sterile solution C, and 10.0mL of sterile solution D. Mix thoroughly. Aseptically and anaerobically distribute into tubes or flasks. Use: For the cultivation and maintenance of Desulfuromonas species. Desulfuromonas spp. Medium (DSMZ Medium 148) Composition per 1001.0mL: Fumaric acid 1.5g KH 2 PO 4 1.0g NH 4 Cl 0.5g Na-acetate 0.5g MgSO 4 ·7H 2 O 0.4g CaCl 2 ·2H 2 O 0.1g Resazurin 0.5mg NaHCO 3 solution 40.0mL Trace elements solution SL-4 10.0mL Na 2 S·9H 2 O solution 6.0mL Vitamin solution 5.0mL pH 7.5 ± 0.2 at 25°C NaHCO 3 Solution: Composition per 100.0mL: NaHCO 3 5.0g Preparation of NaHCO 3 Solution: Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Auto- clave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Must be prepared freshly. Trace Elements Solution SL-4: Composition per liter: EDTA 0.5g FeSO 4 ·7H 2 O 0.2g Trace elements solution SL-6 100.0mL Trace Elements Solution SL-6: Composition per liter: H 3 BO 3 0.3g CoCl 2 ·6H 2 O 0.2g ZnSO 4 ·7H 2 O 0.1g MnCl 2 ·4H 2 O 0.03g Na 2 MoO 4 ·H 2 O 0.03g NiCl 2 ·6H 2 O 0.02g CuCl 2· ·2H 2 O 0.01g Preparation of Trace Elements Solution SL-6: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 3.4. © 2010 by Taylor and Francis Group, LLC Desulfuromonas succinoxidans Medium 579 Preparation of Trace Elements Solution SL-4: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Vitamin Solution: Composition per liter: Pyridoxine-HCl 62.5mg Nicotinic acid 25.0mg Thiamine-HCl·2H 2 O 12.5mg p-Aminobenzoic acid 12.5mg Ca-pantothenate 6.5mg Biotin 2.5mg Preparation of Vitamin Solution: Add components to distilled/ deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize. Na 2 S·9H 2 O Solution: Composition per 100.0mL: Na 2 S·9H 2 O 5.0g Preparation of Na 2 S·9H 2 O Solution: Add Na 2 S·9H 2 O to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Before use, neutralize to pH 7.0 with sterile HCl. Preparation of Medium: Add components, except vitamin solution, NaHCO 3 solution, and Na 2 S·9H 2 O solution, to 950.0mL distilled/deionized water. Adjust pH to 5.0. Autoclave for 15 min at 15 psi pressure– 121°C. Aseptically and anaerobically add 5.0mL vitamin solution, 40.0mL NaHCO 3 solution, and 6.0mL Na 2 S·9H 2 O solution. Mix thoroughly. Adjust pH to 7.5 with sterile 2M Na 2 CO 3 and 2M H 2 SO 4 , re- spectively. Aseptically distribute into sterile screw-capped bottles or tubes. Fill completely, leaving only a small gas bubble. Use: For the cultivation of Desulfuromonas acetexigenes. Desulfuromonas succinoxidans Medium Composition per liter: NaCl 20.0g MgCl 2 ·6H 2 O 2.0g KH 2 PO 4 1.0g MgSO 4 ·7H 2 O 1.0g NH 4 Cl 0.3g CaCl 2 ·2H 2 O 0.1g Resazurin 0.5mg NaHCO 3 solution 50.0mL Disodium fumarate solution 10.0mL Sodium acetate solution 10.0mL Na 2 S·9H 2 O solution 10.0mL Trace elements solution SL-10 1.0mL Seven vitamin solution 1.0mL pH 7.2 ± 0.2 at 25°C NaHCO 3 Solution: Composition per 50.0mL: NaHCO 3 2.5g Preparation of NaHCO 3 Solution: Add NaHCO 3 to distilled/deionized water and bring volume to 50.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure– 121°C. Disodium Fumarate Solution: Composition per 10.0mL: Disodium fumarate 1.6g Preparation of Disodium Fumarate Solution: Add disodium fumarate 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. Sodium Acetate Solution: Composition per 10.0mL: Sodium acetate 0.8g Preparation of Sodium Acetate Solution: Add sodium acetate 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. 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 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 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. Seven Vitamin Solution: Composition per liter: Pyridoxine·HCl 0.3g Thiamine·HCl 0.2g Nicotinic acid 0.2g Calcium DL-pantothenate 0.1g Vitamin B 12 0.1g p-Aminobenzoic acid 80.0mg Biotin 20.0mg Preparation of Seven Vitamin Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize. Sparge with 100% N 2 . Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 . Add components, except NaHCO 3 solution, disodium fumarate solution, sodium acetate solution, Na 2 S·9H 2 O solution, and seven vitamin solution, to distilled/deionized water and bring volume to 920.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Auto- clave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add 50.0mL of sterile NaHCO 3 solution, 10.0mL of sterile disodium fumarate solution, 10.0mL of sterile sodium acetate solution, 10.0mL of sterile Na 2 S·9H 2 O solution, and 1.0mL of sterile seven vitamin solution. Mix thoroughly. Use: For the cultivation of Desulfuromonas succinoxidans. © 2010 by Taylor and Francis Group, LLC 580 Desultobacterium Medium Desultobacterium Medium (DSMZ Medium 383) Composition per 1012.0mL: Soultion A 930.0mL Solution C 50.0mL Solution D 10.0mL Solution E 10.0mL Solution G 10.0mL Solution B 1.0mL Solution F 1.0mL pH 7.0 at 25°C Solution A: Composition per 930.0mL: NaCl 21.0g Na 2 SO 4 3.0g MgCl 2 ·6H 2 O 3.0g KCl 0.5g 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 930.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 gas until saturated. 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 3 300.0mg CuCl 2 ·2H 2 O 2.0mg HCl (25% solution) 7.7mL Preparation of Solution B: 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. Sparge with 100% CO 2 until saturated, approximately 20 min. Filter sterilize under 100% CO 2 into a sterile, gas-tight 100.0mL screw-capped bottle. Solution D: Composition per 10.0mL: Na 2 -benzoate 0.5g Preparation of Solution D: Add Na 2 -benzoate to distilled/deionized water and bring volume to 10.0mL. Sparge with N 2 . Filter sterilize. Store anaerobically. Solution E: 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 Solution E: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Solution F: Composition per liter: NaOH 0.5g Na 2 SeO 3 ·5H 2 O 3.0mg Preparation of Solution F: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Solution G: Composition per 10.0mL: Na 2 S·9H 2 O 0.4g Preparation of Solution G: Add Na 2 S·9H 2 O to distilled/deionized water and bring volume to 10.0mL. Sparge with N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Store anaerobically. Preparation of Medium: Add solutions B, C, D, E, F, and G to solution A in that order under N 2 gas. Adjust the pH to 7.0. Use: For the cultivation of Desulfotobacterium spp. Dethiobacter Medium (DSMZ Medium 1104) Composition per liter: Na 2 CO 3 22.0g NaHCO 3 8.0g NaCl 6.0g Sulfur, powdered 1.5g K 2 HPO 4 0.5g Thiosulfate solution 10.0mL Vitamin solution 10.0mL Ammonium chloride solution 10.0mL Magnesium chloride solution 10.0mL Na 2 S·9H 2 O solution 10.0mL Yeast extract solution 10.0mL Acetate solution 10.0mL Trace elements solution SL-10 with EDTA 1.0mL Selenite/tungstate solution 1.0mL Ethanol 0.6mL pH 9.5 ± 0.2 at 25°C Ammonium Chloride Solution: Composition per 10.0mL: NH 4 Cl 0.2g Preparation of Ammonium Chloride Solution: Add NH 4 Cl 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. Magnesium Chloride Solution: Composition per 10.0mL: MgCl 2 ·6H 2 O 0.2g © 2010 by Taylor and Francis Group, LLC Dethiobacter Medium 581 Preparation of Magnesium Chloride Solution: Add MgCl 2 ·6H 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. Na 2 S·9H 2 O Solution: Composition per 10.0mL: Na 2 S·9H 2 O 0.24g 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. Thiosulfate Solution: Composition per 10.0mL: Na 2 S 2 O 3 2.5g Preparation of Thiosulfate Solution: Add Na 2 S 2 O 3 to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Trace Elements Solution SL-10 with EDTA: 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 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 with EDTA: 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. Adjust pH to 7.0. 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 . 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. Yeast Extract Solution: Composition per 10.0mL: Yeast extract 0.05g 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 . Autoclave for 15 min at 15 psi pressure–121°C. Acetate Solution: Composition per 10.0mL: Sodium acetate 0.4g Preparation of Acetate Solution: Add sodium acetate 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 ammonium chloride, magnesium chloride, yeast extract, trace elements, vitamin, ethanol, acetate, sulfur, thiosulfate, and sulfide solutions, to distilled/ deionized water and bring volume to 930.0mL. Gently heat and bring to boiling. Boil for 1 min. Mix thoroughly. Cool to room temperature while sparging with 100% N 2 . Dispense into culture vessels under an atmosphere of 100% N 2 . Autoclave for 15 min at 15 psi pressure– 121°C. Aseptically add the ammonium chloride, magnesium chloride, yeast extract, trace elements, vitamin, ethanol, acetate, sulfur, thiosulfate, and sulfide solutions. Sulfur is sterilized by steaming for 3 h on each of 3 successive days and added as powder. Adjust pH of final medium to 9.5. After inoculation add sterile H 2 gas to 0.5 bar overpres- sure. Use: For the cultivation of Dethiobacter spp. Dethiobacter Medium (DSMZ Medium 1104) Composition per liter: Na 2 CO 3 22.0g NaHCO 3 8.0g NaCl 6.0g Sulfur, powdered 1.5g K 2 HPO 4 0.5g Thiosulfate solution 10.0mL Vitamin solution 10.0mL Ammonium chloride solution 10.0mL Magnesium chloride solution 10.0mL Na 2 S·9H 2 O solution 10.0mL Yeast extract solution 10.0mL Acetate solution 10.0mL Butyrate solution 10.0mL Trace element ssolution SL-10 with EDTA 1.0mL Selenite/tungstate solution 1.0mL pH 9.5 ± 0.2 at 25°C Ammonium Chloride Solution: Composition per 10.0mL: NH 4 Cl 0.2g Preparation of Ammonium Chloride Solution: Add NH 4 Cl 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. Magnesium Chloride Solution: Composition per 10.0mL: MgCl 2 ·6H 2 O 0.2g Preparation of Magnesium Chloride Solution: Add MgCl 2 ·6H 2 O to distilled/deionized water and bring volume to 10.0mL. Mix thor- © 2010 by Taylor and Francis Group, LLC 582 Dethiosulfovibrio II Medium oughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure– 121°C. Na 2 S·9H 2 O Solution: Composition per 10.0mL: Na 2 S·9H 2 O 0.24g 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. Thiosulfate Solution: Composition per 10.0mL: Na 2 S 2 O 3 2.5g Preparation of Thiosulfate Solution: Add Na 2 S 2 O 3 to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Trace Elements Solution SL-10 with EDTA: 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 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 with EDTA: 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. Adjsut pH to 7.0. 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 . 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. Yeast Extract Solution: Composition per 10.0mL: Yeast extract 0.05g 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 . Autoclave for 15 min at 15 psi pressure–121°C. Acetate Solution: Composition per 10.0mL: Sodium acetate 0.4g Preparation of Acetate Solution: Add sodium acetate 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. Butyrate Solution: Composition per 10.0mL: Sodium butyrate 1.1g Preparation of Butyrate Solution: Add sodium butyrate 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 ammonium chloride, magnesium chloride, yeast extract, trace elements, vitamin, ethanol, acetate, sulfur, thiosulfate, and sulfide solutions, to distilled/ deionized water and bring volume to 920.0mL. Gently heat and bring to boiling. Boil for 1 min. Mix thoroughly. Cool to room temperature while sparging with 100% N 2 . Dispense into culture vessels under an atmosphere of 100% N 2 . Autoclave for 15 min at 15 psi pressure– 121°C. Aseptically add the ammonium chloride, magnesium chloride, yeast extract, trace elements, vitamin, ethanol, acetate, sulfur, thiosulfate, and sulfide solutions. Sulfur is sterilized by steaming for 3 h on each of 3 successive days and added as powder. Adjust pH of final medium to 9.5. After inoculation add sterile H 2 gas to 0.5 bar overpres- sure. Use: For the cultivation of Desulfurivibrio alkaliphilus. Dethiosulfovibrio II Medium (DSMZ Medium 906) Composition per liter: NaCl 20.0g Yeast extract 5.0g MgCl 2 ·6H 2 O 3.0g Na 3 -citrate·5H 2 O 3.0g Peptone 2.0g KH 2 PO 4 1.0g Resazurin 0.5mg Calcium chloride solution 10.0mL Na 2 S·9H 2 O solution 10.0mL Na 2 S 2 O 3 solution 10.0mL Trace elements solution SL-10 1.0mL pH 6.7–6.8 at 25°C 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. Na 2 S·9H 2 O Solution: Composition per 10.0mL: Na 2 S·9H 2 O 0.5g © 2010 by Taylor and Francis Group, LLC DEV Lactose Peptone MUG Broth 583 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. 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 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure– 121°C. Calcium Chloride Solution: Composition per 10.0mL: CaCl 2 ·2H 2 O 0.2g Preparation of Calcium Chloride Solution: Add CaCl 2 ·2H 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. Preparation of Medium: Add components, except calcium chloride solution, trace elements solution SL-10, Na 2 S·9H 2 O solution, and Na 2 S 2 O 3 solution, to distilled/deionized water and bring volume to 969.0mL. Mix thoroughly. Sparge with 100% N 2 for 30 min. Distribute under 100% N 2 into anaerobe tubes or bottles. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically per 1.0L of medium add 10.0mL calcium chloride solution, 1.0mL trace elements solution SL-10, 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 6.7–6.8. Use: For the cultivation of Dethiosulfovibrio spp. Dethiosulfovibrio peptidovorans Medium (DSMZ Medium 786) Composition per 1085.0mL: NaCl 30.0g Trypticase™ 5.0g MgCl 2 ·6H 2 O 3.0g Yeast extract 1.0g NH 4 Cl 1.0g L-Cysteine 0.5g Na-acetate 0.5g K 2 HPO 4 0.3g KH 2 PO 4 0.3g CaCl 2 ·2H 2 O 0.1g KCl 0.1g Resazurin 0.5mg NaHCO 3 solution 50.0mL Na-thiosulfate solution 20.0mL Na 2 S·9H 2 O solution 15.0mL Trace elements solution 10.0mL pH 7.3 ± 0.2 at 25°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 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. Na 2 S·9H 2 O Solution: Composition per 20.0mL: Na 2 S·9H 2 O 0.6g Preparation of Na 2 S·9H 2 O Solution: 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 room temperature. NaHCO 3 Solution: Composition per 100.0mL: NaHCO 3 10.0g Preparation of NaHCO 3 Solution: Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Filter sterilize. Na-thiosulfate Solution: Composition per 20.0mL: Na 2 S 2 O 3 ·5H 2 O 5.0g Preparation of Na-thiosulfate Solution: Add Na 2 S 2 O 3 ·5H 2 O to distilled/deionized water and bring volume to 20.0mL. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2. Add components, except Na-thiosulfate solution, NaHCO 3 solution, and Na 2 S·9H 2 O solution, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Aseptically and anaerobically add 20.0mL Na-thiosulfate solution, 50.0mL NaHCO 3 solution, and 15.0mL Na 2 S·9H 2 O solution. Mix thoroughly. Adjust pH to 7.3. Aseptically and anaerobically distribute into sterile tubes or flasks. Use: For the cultivation of Dethiosulfovibrio peptidovorans. DEV Lactose Peptone MUG Broth Composition per liter: Lactose 10.0g Meat peptone 10.0g NaCl 5.0g Tryptophan 1.0g © 2010 by Taylor and Francis Group, LLC 584 Dextran Agar 4-Methylumbelliferyl-β-D-glucuronide 0.1g Bromocresol Purple 0.01g pH 7.2 ± 0.2 at 37°C Source: This medium is available from Fluka, Sigma-Aldrich. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into test tubes that contain an inverted Durham tube in 10.0mL volumes. Auto- clave for 15 min at 15 psi pressure–121°C. Use: For the enrichment and titer determination of coliform bacteria in connection with the bacteriological examination of water. The pres- ence of E. coli can be demonstrated by fluorescence in the UV and a positive indole test. Dextran Agar Composition per liter: Minimal mineral base solution 700.0mL Agar solution 200.0mL Dextran-deoxyglucose solution 100.0mL pH 4.0 ± 0.2 at 25°C Minimal Mineral Base Solution: Composition per 700.0mL: (NH 4 ) 2 SO 4 5.0g KH 2 PO 4 1.5g CaCl 2 0.1g MnSO 4 0.1g NaCl 0.1g Yeast extract 0.1g Preparation of Minimal Mineral Base Solution: Add components to distilled/deionized water and bring volume to 700.0mL. Mix thoroughly. Gently heat and bring to boiling. pH is 4.0 before autoclav- ing. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Agar Solution: Composition per 200.0mL: Agar 10.0g Preparation of Agar Solution: Add agar to distilled/deionized water and bring volume to 200.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Dextran-Deoxyglucose Solution: Composition per 100.0mL: Dextran 10.0g 2-Deoxy- D-glucose 0.5g Preparation of Dextran-Deoxyglucose Solution: Add components to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Aseptically combine 700.0mL of sterile minimal mineral base solution, 200.0mL of sterile agar solution, and 100.0mL of sterile dextran-deoxyglucose solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of Lipomyces starkeyi. Dextran Broth Composition per liter: Minimal mineral base solution 900.0mL Dextran-deoxyglucose solution 100.0mL pH 4.0 ± 0.2 at 25°C Minimal Mineral Base Solution: Composition per 900.0mL: (NH 4 ) 2 SO 4 5.0g KH 2 PO 4 1.5g CaCl 2 0.1g MnSO 4 0.1g NaCl 0.1g Yeast extract 0.1g Preparation of Minimal Mineral Base Solution: Add components to distilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Gently heat and bring to boiling. pH is 4.0 before autoclav- ing. Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature. Dextran-Deoxyglucose Solution: Composition per 100.0mL: Dextran 10.0g 2-Deoxy- D-glucose 0.5g Preparation of Dextran-Deoxyglucose Solution: Add components to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Combine 900.0mL of sterile minimal mineral base solution with 100.0mL of sterile dextran-deoxyglucose solution. Mix thoroughly. Distribute into sterile flasks or tubes. Use: For the cultivation and maintenance of Lipomyces starkeyi. Dextran Medium (DSMZ Medium 1050) Composition per liter: Agar 15.0g Dextran 10.0g Polypepton 5.0g NH 4 NO 3 1.0g KH 2 PO 4 1.0g MgSO 4 ·7H 2 O 0.5g CaCl 2 ·2H 2 O 0.25g pH 6.8 ± 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 6.8. Gently heat while stirring 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 Arthrobacter dextranyliticum. Dextrin Fuchsin Sulfite Agar See: Aeromonas Differential Agar Dextrose Agar Composition per liter: Agar 15.0g Glucose 10.0g NaCl 5.0g Pancreatic digest of casein 5.0g Peptic digest of animal tissue 5.0g Beef extract 3.0g pH 6.9 ± 0.2 at 25°C Source: This medium is available as a premixed powder from BD Di- agnostic Systems. © 2010 by Taylor and Francis Group, LLC . slurry. Preparation of Medium: To 1.0L of cooled, sterile solution 1, aseptically add 1.0mL of sterile solution 2, 40.0mL of sterile solution 3, 6.0mL of sterile solution 4, and 5.0mL of sterile solution. slurry. Preparation of Medium: To 1.0L of cooled, sterile solution 1, aseptically add 1.0mL of sterile solution 2, 40.0mL of sterile solution 3, 6.0mL of sterile solution 4, and 5.0mL of sterile solution. add 50.0mL of sterile NaHCO 3 solution, 10.0mL of sterile disodium fumarate solution, 10.0mL of sterile sodium acetate solution, 10.0mL of sterile Na 2 S·9H 2 O solution, and 1.0mL of sterile