Handbook of Microbiological Media, Fourth Edition part 102 pps

Mannitol Agar with Prilion 1005 tion. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the isolation and cultivation of iron and sulfur bacteria. Also used to differentiate Leptothrix (Sphaerotilus) discophorus from Spha- erotilus natans. Manganese Agar No. 2 (Mn Agar No. 2) Composition per liter: Agar 15.0g MnSO 4 ·H 2 O 10.0mg 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 freshly prepared solution. Use: For the enumeration, enrichment, and isolation of iron and sulfur bacteria. For the isolation and cultivation of Leptothrix species from water. Manganese Medium for Pseudomonas species Composition per liter: Noble agar 10.0g MnCO 3 1.0g Fe(NH 4 ) 2 (SO 4 ) 2 ·6H 2 O 0.15g Sodium citrate 0.15g Yeast extract 0.075g Na 4 P 2 O 7 ·10H 2 O 0.05g 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. 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 Pseudomonas putida and other Pseudomonas species. Manganese Nutrient Agar Composition per liter: Agar 15.0g Peptone 5.0g Meat extract 3.0g MnSO 4 ·H 2 O 5.0mg 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 obtaining the sporulation of Bacillus species. Manning Medium (DSMZ Medium 1023) Composition per liter: (NH 4 ) 2 SO 4 6.0g MgSO 4 ·7H 2 O 1.0g KCl 0.2g K 2 HPO 4 0.2g Ca(NO 3 ) 2 0.02g Solution A 100.0mL Solution B 10.0mL pH 2.6 ± 0.1 at 25°C Solution A: Composition per 100.0mL: FeSO 4 ·7H 2 O 33.4g Preparation of Solution A: Add FeSO 4 ·7H 2 O to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Solution B: Composition per 10.0mL: Yeast extract 0.2g Preparation of Solution B: Add yeast extract to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: Add components, except solutions A and B, to distilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Adjust pH to 2.6. Gently heat while stirring and bring to boiling. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Asepti- cally add solutions A and B. Adjust pH to 2.5–2.7 with 0.1N H 2 SO 4 . Use: For the cultivation of iron-oxidizing and heterotrophic acido- philic bacteria from acid mine drainage. Mannitol Agar Composition per liter: Mannitol 25.0g Agar 15.0g Yeast extract 5.0g Peptone 3.0g Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Acetobacter aceti, Aceto- bacter hansenii, Acetobacter pasteurianus, Frateuria aurantia, Glu- conobacter asaii, Gluconobacter cerinus, Gluconobacter oxydans, and other bacteria that can utilize mannitol as a carbon source. Mannitol Agar with Prilion Composition per liter: D-Mannitol 15.0g Agar 13.0g Meat peptone 10.0g Meat extract 7.0g NaCl 3.0g Na 2 H 2 PO 4 2.0g Prilion 2.0g Metachrome Yellow 1.875g Water Blue 0.625g pH 7.2 ± 0.2 at 25°C Source: This medium is available from HiMedia. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the isolation and differentiation of Salmonella spp. from Pro- teus species. © 2010 by Taylor and Francis Group, LLC 1006 Mannitol Egg Yolk Polymyxin Agar Mannitol Egg Yolk Polymyxin Agar Composition per liter: Agar 15.0g D-Mannitol 10.0g Peptone 10.0g NaCl 10.0g Beef extract 1.0g Phenol Red 0.025g Egg yolk emulsion, 50% 50.0mL Polymyxin B solution 10.0mL pH 7.1 ± 0.1 at 25°C Source: Available as a prepared medium from BD Diagnostic Sys- tems. Egg Yolk Emulsion, 50%: Composition per 100.0mL: Chicken egg yolks 11 Whole chicken egg 1 NaCl (0.9% solution) 80.0mL Preparation of Egg Yolk Emulsion, 50%: Soak eggs with 1:100 dilution of saturated mercuric chloride solution for 1 min. Crack eggs and separate yolks from whites. Mix egg yolks with 1 chicken egg. Beat to form emulsion. Measure 50.0mL of egg yolk emulsion and add to 50.0mL of 0.9% NaCl solution. Mix thoroughly. Filter sterilize. Warm to 45°–50°C. Polymyxin B Solution: Composition per 10.0mL: Polymyxin B 100,000U Preparation of Polymyxin B Solution: Add polymyxin B to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components—except egg yolk emulsion, 50%, and polymyxin B solution—to distilled/deionized water and bring volume to 940.0mL. 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 50.0mL of sterile egg yolk emulsion, 50%, and 10.0mL of sterile polymyxin B solution. Mix thoroughly. Pour into sterile Petri dishes. Use: For the cultivation and enumeration of Bacillus cereus from foods. Mannitol Lysine Crystal Violet Brilliant Green Agar See: MLCB Agar Mannitol Maltose Agar Composition per liter: NaCl 20.0g Agar 13.0g D-Mannitol 10.0g Maltose 10.0g Beef extract 5.0g Papaic digest of soybean meal 5.0g Polypeptone™ 5.0g Dye stock solution, 1000X 1.0mL pH 7.8 ± 0.2 at 25°C Dye Stock Solution, 1000X: Composition per 100.0mL: Bromthymol Blue 4.0g Cresol Red 4.0g Ethanol, 95% 100.0mL Preparation of Dye Stock Solution, 1000X: Add Bromthymol Blue and Cresol Red to 100.0mL of ethanol. Mix thoroughly. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust to pH 7.8. Gently heat and bring to boiling. Distribute into tubes or flasks. Auto- clave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation of Vibrio species from foods. Mannitol Motility Test HiVeg Medium Composition per liter: Plant peptone 20.0g Agar 3.0g Mannitol 2.0g Potassium nitrate 1.0g Phenol red 0.04g pH 7.6 ± 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 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 selective isolation, cultivation, and enumeration of staphylococci from clinical and nonclinical specimens. Mannitol-utilizing organisms turn the medium yellow. Mannitol Salt Agar Composition per liter: NaCl 75.0g Agar 15.0g D-Mannitol 10.0g Pancreatic digest of casein 5.0g Peptic digest of animal tissue 5.0g Beef extract 1.0g Phenol Red 0.025g pH 7.4 ± 0.2 at 25°C Source: This medium is available as a premixed powder from BD Di- agnostic Systems and Oxoid Unipath. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat 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 selective isolation, cultivation, and enumeration of staphylococci from clinical and nonclinical specimens. Mannitol-utilizing organisms turn the medium yellow. Mannitol Salt Agar (BAM M97) Composition per liter: NaCl 75.0g Agar 15.0g D-Mannitol 10.0g Polypeptone 10.0g © 2010 by Taylor and Francis Group, LLC Mannitol Selenite Broth 1007 Beef extract 1.0g Phenol Red 0.025g pH 7.4 ± 0.2 at 25°C Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat 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 selective isolation, cultivation, and enumeration of staphylococci. Mannitol-utilizing organisms turn the medium yellow. Mannitol Salt Agar with Egg Yolk Emulsion Composition per liter: NaCl 75.0g Agar 15.0g D-Mannitol 10.0g Proteose peptone 10.0g Beef extract 1.0g Phenol Red 0.025g Egg yolk emulsion 100.0mL pH 7.4 ± 0.2 at 25°C Source: This medium is available from HiMedia. Egg Yolk Emulsion Composition per 100.0mL: Chicken egg yolks 11 Whole chicken egg 1 NaCl (0.9% solution) 50.0mL Preparation of Egg Yolk Emulsion: Soak eggs with 1:100 dilution of saturated mercuric chloride solution for 1 min. Crack eggs and separate yolks from whites. Mix egg yolks with 1 chicken egg. Beat to form emulsion. Measure 50.0mL of egg yolk emulsion and add to 50.0mL of 0.9% NaCl solution. Mix thoroughly. Filter sterilize. Warm to 45°–50°C. Preparation of Medium: Add components, except egg yolk emulsion, to distilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Aseptically add egg yolk emulsion. Mix thoroughly. Pour into Petri dishes or aseptically distribute into sterile tubes or flasks. Use: For the selective isolation of pathogenic staphylococci. Mannitol Salt Broth Composition per liter: NaCl 75.0g Proteose peptone 10.0g D-Mannitol 10.0g Beef extract 1.0g Phenol Red 0.025g pH 7.4 ± 0.2 at 25°C Source: This medium is available from HiMedia. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the selective isolation of presumptive pathogenic staphylococci. Mannitol Salt Broth Composition per liter: NaCl 100.0g Pancreatic digest of casein 17.0g Papaic digest of soybean meal 3.0g K 2 HPO 4 2.5g D-Mannitol 2.5g Phenol Red 0.025g pH 7.4 ± 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. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the selective isolation and cultivation of staphylococci from foods and nonclinical specimens. Mannitol-utilizing organisms turn the medium yellow. Mannitol Salt HiVeg Agar Base Composition per liter: NaCl 75.0g Agar 15.0g D-Mannitol 10.0g Plant peptone No. 3 10.0g Plant extract 1.0g Phenol Red 0.025g pH 7.4 ± 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 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 selective isolation of pathogenic staphylococci. Mannitol Salt HiVeg Broth Composition per liter: NaCl 75.0g D-Mannitol 10.0g Plant peptone No. 3 10.0g Plant extract 1.0g Phenol Red 0.025g pH 7.4 ± 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 while stirring and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the selective isolation of presumptive pathogenic staphylococci. Mannitol Selenite Broth (Selenite Mannitol Broth) Composition per liter: NaH 2 PO 4 10.0g Peptic digest of animal tissue 5.0g © 2010 by Taylor and Francis Group, LLC 1008 Mannitol Selenite Broth with Brilliant Green Mannitol 4.0g NaHSeO 3 4.0g pH 7.1 ± 0.2 at 25°C Source: This medium is available from HiMedia. Caution: Sodium hydrogen selenite (sodium biselenite) is a very tox- ic, corrosive agent and causes teratogenicity; it should be handled with great care. If there is contact, wash immediately with lots of water. Preparation of Medium: Add NaHSeO 3 to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Add remaining components. Mix thoroughly. Distribute into tubes or flasks. Gently heat and bring to boiling. Sterilize in a boiling water bath or free flowing steam for 10 min. Do not autoclave. Discard the prepared medium if a large amount of selenite is reduced (indicated by red precipitate at the bot- tom of the tube). Use: For the selective enrichment of Salmonella spp. from clinical materials. Mannitol Selenite Broth with Brilliant Green Composition per liter: Meat peptone 5.0g Yeast extract 5.0g Mannitol 5.0g K 2 HPO 4 4.35g KH 2 PO 4 3.4g Sodium taurocholate 1.0g Brilliant Green 0.005g Na 2 SeO 3 ·5H 2 O 1.0g pH 7.0 ± 0.2 at 25°C Source: This medium is available from HiMedia. Caution: Sodium hydrogen selenite (sodium biselenite) is a very tox- ic, corrosive agent and causes teratogenicity; iit should be handled with great care. If there is contact, wash immediately with lots of water. Preparation of Medium: Add NaHSeO 3 to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Add remaining components. Mix thoroughly. Distribute into tubes or flasks. Gently heat and bring to boiling. Sterilize in a boiling water bath or free flowing steam for 10 min. Do not autoclave. Discard the prepared medium if a large amount of selenite is reduced (indicated by red precipitate at the bot- tom of the tube). Use: For the enrichment of Salmonella spp. from feces, foodstuffs, and other materials. Mannitol Yeast Extract Medium (LMG 135) Composition per liter: Agar 20.0g Mannitol 10.0g Yeast extract 1.0g KH 2 PO 4 0.5g NaCl 0.1g CaCl 2 ·2H 2 O solution 1.0mL FeCl 3 ·6H 2 O solution 1.0mL CaCl 2 ·2H 2 O Solution: Composition per 10.0mL: CaCl 2 ·2H 2 O 5.28mg Preparation of CaCl 2 ·2H 2 O Solution: Add CaCl 2 ·2H 2 O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. FeCl 3 ·6H 2 O Solution: Composition per 10.0mL: FeCl 3 ·6H 2 O 0.66mg Preparation of FeCl 3 ·6H 2 O Solution: Add FeCl 3 ·6H 2 O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. 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 Rhizobium species and Bradyrhizobium species. Mannitol-Yeast Extract-Peptone (MYP) (DSMZ Medium 1087) Composition per liter: D-Mannitol 25.0g Agar 15.0g Yeast extract 5.0g Peptone 3.0g pH 7.2 ± 0.2 at 25°C Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.2. 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 Acetobacter fabarum. Mannitol Yolk Polymyxin Agar (MYP Agar) Composition per 110.0mL: Agar 1.5g NaCl 1.0g Peptone 1.0g D-Mannitol 1.0g (NH 4 ) 2 PO 4 0.1g Meat extract 0.1g Phenol Red 2.5mg Egg yolk emulsion, 20% 10.0mL Polymyxin B solution 1.0mL pH 7.1 ± 0.2 at 25°C Egg Yolk Emulsion, 20%: Composition per 100.0mL: Chicken egg yolks 11 Whole chicken egg 1 NaCl (0.9% solution) 80.0mL Preparation of Egg Yolk Emulsion, 20%: Soak eggs with 1:100 dilution of saturated mercuric chloride solution for 1 min. Crack eggs and separate yolks from whites. Mix egg yolks with 1 chicken egg. Measure 20.0mL of egg yolk emulsion and add to 80.0mL of 0.9% NaCl solution. Mix thoroughly. Filter sterilize. Warm to 45°–50°C. Polymyxin B Solution: Composition per 1.0mL: Polymyxin B 1.0mg Preparation of Polymyxin B Solution: Add polymyxin B to distilled/deionized water and bring volume to 1.0mL. Mix thoroughly. Fil- ter sterilize. © 2010 by Taylor and Francis Group, LLC Marine Agar 2216 1009 Preparation of Medium: Add components—except egg yolk emulsion, 20%, and polymyxin B solution—to distilled/deionized water and bring volume to 100.0mL. 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 10.0mL of sterile egg yolk emulsion, 20%, and 1.0mL of sterile polymyxin B solution. Mix thoroughly. Pour into sterile Petri dishes. Use: For the cultivation and maintenance of Bacillus cereus. Maricaulis Medium (DSMZ Medium 1025) Composition per liter: Sea salts, Sigma 30.0g NH 4 Cl 0.5g Peptone yeast extract solution 20.0mL Glucose solution 2.0mL Riboflavin solution 5.0mL pH 7.2 ± 0.2 at 25°C Glucose Solution: Composition per 10.0mL: Glucose 5.0g Preparation of Glucose Solution: Add glucose to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Riboflavin Solution: Composition per 10.0mL: Riboflavin 2.0mg Preparation of Riboflavin Solution: Add riboflavin to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Peptone Yeast Extract Solution: Composition per 100.0mL: Peptone 10.0g Yeast extract 5.0g Preparation of Peptone Yeast Extract Solution: Add components to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: Add components, except riboflavin, glucose, and peptone yeast extract solutions, to distilled/deionized water and bring volume to 980.0mL. Mix thoroughly. Adjust pH to 7.2. Gen- tly heat while stirring and bring to boiling. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature. Asep- tically add riboflavin, glucose, and peptone yeast extract solutions. Use: For the cultivation of Maricaulis spp. Marine Agar (DSMZ Medium 123) Composition per liter: Agar 15.0g Tryptone 10.0g Peptone 5.0g Yeast extract 1.0g Synthetic seawater 1.0L pH 7.8 ± 0.2 at 25°C Synthetic Seawater: Composition per liter: NaCl 24.0g MgCl 2 ·6H 2 O 11.0g Na 2 SO 4 4.0g CaCl 2 ·6H 2 O 2.0g KCl 0.7g KBr 0.1g SrCl 2 ·6H 2 O 0.04g H 3 BO 3 0.03g NaSiO 3 ·9H 2 O 5.0mg NaF 3.0mg NH 4 NO 3 2.0mg Fe 3 PO 4 ·4H 2 O 1.0mg Preparation of Synthetic Seawater: Add components to distilled water and bring volume to 1.0L. Mix thoroughly. Preparation of Medium: Add agar, tryptone, peptone, and yeast extract to synthetic seawater and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.8. 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 Halobacillus halophilus, Halomonas spp., Vibrio harveyi, Cobetia marina, and Ruegeria atlantica. Marine Agar 2216 (DSMZ Medium 604) Composition per liter: NaCl 19.45g Agar 15.0g MgCl 2 8.8g Peptone 5.0g Na 2 SO 3 3.24g CaCl 2 1.8g Yeast extract 1.0g KCl 0.55g NaHCO 3 0.16g Ferric citrate 0.1g KBr 0.08g SrCl 2 0.03g H 3 BO 3 0.02g Na 2 HPO 4 8.0mg Na 2 SiO 3 4.0mg NaF 2.4mg NH 4 NO 3 1.6mg pH 7.6 ± 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 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 and maintenance of Hyphomonas spp., Ocean- ospirillum spp., Hyphomicrobium indicum, Psychroflexus gondwanen- sis=Flavobacterium gondwanense, Salegentibacter salegens=Fla- vobacterium salegens, Psychromonas antarctica, Sulfitobacter mediterraneus, Thalassomonas viridans, Vibrio spp., Marinospirillum minutulum=Oceanospirillum minutulum, Terasakiella pusilla=Oceano- spirillum pusillum, Pseudoalteromonas atlantica=Alteromonas atlantica, Pseudomonas atlantica, Roseobacter spp., Erythrobacter longus, Pseudospirillum japonicum=Oceanospirillum japonicum, Marino- bacter hydrocarbonoclasticus (Pseudomonas nautica), Psychrobacter © 2010 by Taylor and Francis Group, LLC 1010 Marine Agar with Biphenyl spp., and Moritella japonica. For the isolation, cultivation, and maintenance of a wide variety of heterotrophic marine bacteria. Marine Agar with Biphenyl Composition per liter: NaCl 19.45g Agar 15.0g MgCl 2 8.8g Peptone 5.0g Na 2 SO 3 3.24g CaCl 2 1.8g Yeast extract 1.0g KCl 0.55g NaHCO 3 0.16g Ferric citrate 0.1g KBr 0.08g SrCl 2 0.03g H 3 BO 3 0.02g Na 2 HPO 4 8.0mg Na 2 SiO 3 4.0mg NaF 2.4mg NH 4 NO 3 1.6mg Biphenyl 1.0mg pH 7.6 ± 0.2 at 25°C Preparation of Medium: Add components, except biphenyl, 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. After agar solidifies, aseptically add a few crystals of biphenyl to each plate. Use: For the cultivation and maintenance of biphenyl-utilizing marine bacteria, such as Cycloclasticus pugetii. Marine Agar with Lambda Carrageenan Composition per 1070.0mL: Solution A 1.0L Solution B 60.0mL Solution C 10.0mL pH 7.2 ± 0.2 at 25°C Solution A: Composition per liter: NaCl 25.0g Agar 15.0g MgSO 4 ·7H 2 O 5.0g Casamino acids 2.5g Lambda-carrageenan 2.5g NaNO 3 2.0g CaCl 2 ·2H 2 O 0.2g KCl 0.1g Preparation of Solution A: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Solution B: Composition per 100.0mL: Na 2 HPO 4 ·2H 2 O 3.56g Preparation of Solution B: Add Na 2 HPO 4 ·2H 2 O to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Auto- clave for 15 min at 15 psi pressure–121°C. Solution C: Composition per 100.0mL: FeSO 4 ·7H 2 O 0.3g Preparation of Solution C: Add FeSO 4 ·7H 2 O to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: Aseptically add 60.0mL of sterile solution B and 10.0mL of sterile solution C to 1.0L of sterile solution A. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of ATCC strain 43554. Marine Agar with Kappa and Lambda Carrageenan Composition per 1070.0mL: Solution A 1.0L Solution B 60.0mL Solution C 10.0mL pH 7.2 ± 0.2 at 25°C Solution A: Composition per liter: NaCl 25.0g Agar 15.0g MgSO 4 ·7H 2 O 5.0g Casamino acids 2.5g NaNO 3 2.0g κ-Carrageenan 1.25g λ-Carrageenan 1.25g CaCl 2 ·2H 2 O 0.2g KCl 0.1g Preparation of Solution A: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Solution B: Composition per 100.0mL: Na 2 HPO 4 ·2H 2 O 3.56g Preparation of Solution B: Add Na 2 HPO 4 ·2H 2 O to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Auto- clave for 15 min at 15 psi pressure–121°C. Solution C: Composition per 100.0mL: FeSO 4 ·7H 2 O 0.3g Preparation of Solution C: Add FeSO 4 ·7H 2 O to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: Aseptically add 60.0mL of sterile solution B and 10.0mL of sterile solution C to 1.0L of sterile solution A. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of Pseudomonas carrageen- ovora. Marine Agar with Naphthalene Composition per liter: NaCl 19.45g Agar 15.0g MgCl 2 8.8g © 2010 by Taylor and Francis Group, LLC Marine Broth with Biphenyl 1011 Peptone 5.0g Na 2 SO 3 3.24g CaCl 2 1.8g Yeast extract 1.0g KCl 0.55g NaHCO 3 0.16g Ferric citrate 0.1g KBr 0.08g SrCl 2 0.03g H 3 BO 3 0.02g Na 2 HPO 4 8.0mg Na 2 SiO 3 4.0mg NaF 2.4mg NH 4 NO 3 1.6mg Naphthalene 1mg pH 7.6 ± 0.2 at 25°C Preparation of Medium: Add components, except naphthalene, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Auto- clave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. After agar solidifies, aseptically add a few crystals of naphthalene to each plate. Use: For the cultivation and maintenance of naphthalene-utilizing marine bacteria Marine Agar with Sulfur (ATCC Medium 1922) Composition per liter: NaCl 19.45g Sulfur 10.0g MgCl 2 8.8g Peptone 5.0g Na 2 SO 3 3.24g CaCl 2 1.8g Yeast extract 1.0g KCl 0.55g NaHCO 3 0.16g Ferric citrate 0.1g KBr 0.08g SrCl 2 0.03g H 3 BO 3 0.02g Na 2 HPO 4 8.0mg Na 2 SiO 3 4.0mg NaF 2.4mg NH 4 NO 3 1.6mg pH 7.6 ± 0.2 at 25°C Preparation of Sulfur: Autoclave sulfur for 15 min at 0 psi pressure– 100°C on 3 successive days. Preparation of Medium: Prepare anaerobically under a gas phase of 80% N 2 + 10% CO 2 + 10% H 2 . Add components, except sulfur, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat while stirring and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50–55°C. Aseptically add 10.0g of sterile sulfur. Mix thoroughly. Aseptically and anaerobically, under a gas phase of 80% N 2 + 10% CO 2 + 10% H 2 , distribute into sterile tubes. Use: For the cultivation and maintenance of Thermococcus litoralis. Marine Ameba Medium Composition per liter: Agar 10.0g Malt extract 0.1g Yeast extract 0.1g Artificial seawater 1.0L Artificial Seawater: Composition per liter: NaCl 27.5g MgSO 4 ·7H 2 O 6.78g MgCl 2 ·6H 2 O 5.38g KCl 0.72g NaHCO 3 0.2g CaCL 2 ·2H 2 O 1.4g Preparation of Artificial Seawater: Add components to distilled/ deionized water and bring volume to 1.0L. Mix thoroughly. Preparation of Medium: Add components to artificial seawater and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation of Cochliopodium clarum, Heteramoeba clara, Lingulamoeba leei, Paramoeba pemaquidensis, and Vannella species. Marine Broth 2216 (LMG Medium 164) Composition per liter: NaCl 19.45g MgCl 2 8.8g Peptone 5.0g Na 2 SO 3 3.24g CaCl 2 1.8g Yeast extract 1.0g KCl 0.55g NaHCO 3 0.16g Ferric citrate 0.1g KBr 0.08g SrCl 2 0.03g H 3 BO 3 0.02g Na 2 HPO 4 8.0mg Na 2 SiO 3 4.0mg NaF 2.4mg NH 4 NO 3 1.6mg pH 7.6 ± 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 while stirring and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of Vibrio liquefaciens and for the isolation, cultivation, and maintenance of a wide variety of heterotrophic marine bacteria. Marine Broth with Biphenyl Composition per liter: NaCl 19.45g MgCl 2 8.8g © 2010 by Taylor and Francis Group, LLC 1012 Marine Broth with Lambda Carrageenan Peptone 5.0g Na 2 SO 3 3.24g CaCl 2 1.8g Yeast extract 1.0g KCl 0.55g NaHCO 3 0.16g Ferric citrate 0.1g KBr 0.08g SrCl 2 0.03g H 3 BO 3 0.02g Na 2 HPO 4 8.0mg Na 2 SiO 3 4.0mg NaF 2.4mg NH 4 NO 3 1.6mg Biphenyl 1.0mg pH 7.6 ± 0.2 at 25°C Preparation of Medium: Add components, except biphenyl, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Dis- tribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure– 121°C. Aseptically add a few crytals of biphenyl to each tube or flask. Use: For the cultivation of biphenyl-utilizing marine bacteria. Marine Broth with Lambda Carrageenan Composition per 1070.0mL: Solution A 1.0L Solution B 60.0mL Solution C 10.0mL pH 7.2 ± 0.2 at 25°C Solution A: Composition per liter: NaCl 25.0g MgSO 4 ·7H 2 O 5.0g Casamino acids 2.5g λ-Carrageenan 2.5g NaNO 3 2.0g CaCl 2 ·2H 2 O 0.2g KCl 0.1g Preparation of Solution A: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Solution B: Composition per 100.0mL: Na 2 HPO 4 ·2H 2 O 3.56g Preparation of Solution B: Add Na 2 HPO 4 ·2H 2 O to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Auto- clave for 15 min at 15 psi pressure–121°C. Solution C: Composition per 100.0mL: FeSO 4 ·7H 2 O 0.3g Preparation of Solution C: Add FeSO 4 ·7H 2 O to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: Aseptically add 60.0mL of sterile solution B and 10.0mL of sterile solution C to 1.0L of sterile solution A. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation of ATCC strain 43554. Marine Broth with Kappa and Lambda Carrageenan Composition per 1070.0mL: Solution A 1.0L Solution B 60.0mL Solution C 10.0mL pH 7.2 ± 0.2 at 25°C Solution A: Composition per liter: NaCl 25.0g MgSO 4 ·7H 2 O 5.0g Casamino acids 2.5g NaNO 3 2.0g κ-Carrageenan 1.25g λ-Carrageenan 1.25g CaCl 2 ·2H 2 O 0.2g KCl 0.1g Preparation of Solution A: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Solution B: Composition per 100.0mL: Na 2 HPO 4 ·2H 2 O 3.56g Preparation of Solution B: Add Na 2 HPO 4 ·2H 2 O to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Auto- clave for 15 min at 15 psi pressure–121°C. Solution C: Composition per 100.0mL: FeSO 4 ·7H 2 O 0.3g Preparation of Solution C: Add FeSO 4 ·7H 2 O to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: Aseptically add 60.0mL of sterile solution B and 10.0mL of sterile solution C to 1.0L of sterile solution A. Mix thoroughly. Distribute into sterile tubes or flasks. Use: For the cultivation and maintenance of Pseudomonas carrageen- ovora. Marine Broth with Naphthalene Composition per liter: NaCl 19.45g MgCl 2 8.8g Peptone 5.0g Na 2 SO 3 3.24g CaCl 2 1.8g Yeast extract 1.0g KCl 0.55g NaHCO 3 0.16g Ferric citrate 0.1g KBr 0.08g SrCl 2 0.03g H 3 BO 3 0.02g Na 2 HPO 4 8.0mg Na 2 SiO 3 4.0mg NaF 2.4mg NH 4 NO 3 1.6mg Naphthalene 1mg pH 7.6 ± 0.2 at 25°C © 2010 by Taylor and Francis Group, LLC Marine Chlorobiaceae Medium 2 1013 Preparation of Medium: Add components, except naphthalene, 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. Aseptically add a few crytals of naphthalene to each tube or flask. Use: For the cultivation of naphthalene-utilizing marine bacteria. Marine Broth with Sulfur Composition per liter: NaCl 19.45g Sulfur 10.0g MgCl 2 8.8g Peptone 5.0g Na 2 SO 3 3.24g CaCl 2 1.8g Yeast extract 1.0g KCl 0.55g NaHCO 3 0.16g Ferric citrate 0.1g KBr 0.08g SrCl 2 0.03g H 3 BO 3 0.02g Na 2 HPO 4 8.0mg Na 2 SiO 3 4.0mg NaF 2.4mg NH 4 NO 3 1.6mg pH 7.6 ± 0.2 at 25°C Preparation of Sulfur: Autoclave for 15 min at 0 psi pressure–100°C on three successive days. Preparation of Medium: Prepare anaerobically under a gas phase of 80% N 2 + 10% CO 2 + 10% H 2 . Add components, except sulfur, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat while stirring and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Aseptically add 10.0g of sulfur. Mix thoroughly. Aseptically and anaerobically, under a gas phase of 80% N 2 + 10% CO 2 + 10% H 2 , distribute into sterile tubes. Use: For the cultivation of Thermococcus litoralis. Marine Caulobacter Medium Composition per liter: Proteose peptone 10.0g Yeast extract 3.0g Artificial seawater 1.0L pH 7.2–7.4 at 25°C Artificial Seawater: Composition per liter: Commercially available marine aquarium salts mixture variable Preparation of Artificial Seawater: Add commercially available marine aquarium salts mixture to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Preparation of Medium: Combine components. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure– 121°C. Use: For the cultivation of Caulobacter halobacteroides and Cau- lobacter maris. Marine Chlorobiaceae Medium 2 Composition per 1051.0mL: Solution 1 950.0mL Na 2 S·9H 2 O solution 60.0mL NaHCO 3 solution 40.0mL Vitamin B 12 solution 1.0mL pH 6.8 ± 0.2 at 25°C Solution 1: Composition per 950.0mL: NaCl 20.0g MgSO 4 ·7H 2 O 3.0g KH 2 PO 4 1.0g NH 4 Cl 0.5g CaCl 2 ·2H 2 O 0.05g Trace elements solution SL-8 1.0mL Preparation of Solution 1: Add components to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Trace Elements Solution SL-8: Composition per liter: Disodium EDTA 5.2g FeCl 2 ·4H 2 O 1.5g CoCl 2 ·6H 2 O 0.19g MnCl 2 ·4H 2 O 0.1g ZnCl 2 0.07g H 3 BO 3 0.06g NaMoO 4 ·2H 2 O 0.04g CuCl 2 ·2H 2 O 0.02g NiCl 2 ·6H 2 O 0.02g Preparation of Trace Elements Solution SL-8: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. 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. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°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. Filter sterilize. Vitamin B 12 Solution: Composition per 100.0mL: Vitamin B 12 2.0mg Preparation of Vitamin B 12 Solution: Add vitamin B 12 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: To 950.0mL of cooled, sterile solution 1, aseptically add 60.0mL of sterile Na 2 S·9H 2 O solution, 40.0mL of sterile NaHCO 3 solution, and 1.0mL of sterile vitamin B 12 solution. Mix thoroughly. Adjust pH to 6.8 with sterile H 2 SO 4 or Na 2 CO 3 . Aseptical- ly distribute into sterile 50.0mL or 100.0mL bottles with metal screw caps and rubber seals. Completely fill bottles with medium except for a pea-sized air bubble. © 2010 by Taylor and Francis Group, LLC 1014 Marine Chromatiaceae Medium 2 Use: For the isolation and cultivation of marine members of the Chlo- robiaceae. Marine Chromatiaceae Medium 2 Composition per 1051.0mL: Solution 1 950.0mL Na 2 S·9H 2 O solution 60.0mL NaHCO 3 solution 40.0mL Vitamin B 12 solution 1.0mL pH 7.3 ± 0.2 at 25°C Solution 1: Composition per 950.0mL: NaCl 20.0g MgSO 4 ·7H 2 O 3.0g KH 2 PO 4 1.0g NH 4 Cl 0.5g CaCl 2 ·2H 2 O 0.05g Trace elements solution SL-8 1.0mL Preparation of Solution 1: Add components to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Trace Elements Solution SL-8: Composition per liter: Disodium EDTA 5.2g FeCl 2 ·4H 2 O 1.5g CoCl 2 ·6H 2 O 0.19g MnCl 2 ·4H 2 O 0.1g ZnCl 2 0.07g H 3 BO 3 0.06g NaMoO 4 ·2H 2 O 0.04g CuCl 2 ·2H 2 O 0.02g NiCl 2 ·6H 2 O 0.02g Preparation of Trace Elements Solution SL-8: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. 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. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°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. Filter sterilize. Vitamin B 12 Solution: Composition per 100.0mL: Vitamin B 12 2.0mg Preparation of Vitamin B 12 Solution: Add vitamin B 12 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: To 950.0mL of cooled, sterile solution 1, aseptically add 60.0mL of sterile Na 2 S·9H 2 O solution, 40.0mL of sterile NaHCO 3 solution, and 1.0mL of sterile vitamin B 12 solution. Mix thoroughly. Adjust pH to 7.3 with sterile H 2 SO 4 or Na 2 CO 3 . Aseptical- ly distribute into sterile 50.0mL or 100.0mL bottles with metal screw caps and rubber seals. Completely fill bottles with medium except for a pea-sized air bubble. Use: For the isolation and cultivation of marine members of the Chro- matiaceae. Marine Cytophaga Agar Composition per liter: Agar 15.0g Nutrient broth 8.0g Yeast extract 5.0g Salt solution 1.0L Salt Solution: Composition per liter: NaCl 12.86g MgCl 2 2.48g KCl 0.75g CaCl 2 0.56g Fe(SO 4 ) 2 (NH 4 ) 2 0.048g Preparation of Salt Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Preparation of Medium: Add solid components to 1.0L of salt solution. Mix thoroughly. Gently heat while stirring 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 Cytophaga species. Marine Cytophaga Medium Composition per liter: NaCl 24.7g Agar 15.0g MgSO 4 ·7H 2 O 6.3g MgCl 2 ·6H 2 O 4.6g Tryptic digest of casein 1.0g Yeast extract 1.0g KCl 0.7g NaHCO 3 solution 10.0mL CaCl 2 ·2H 2 O solution 10.0mL pH 7.2 ± 0.2 at 25°C NaHCO 3 Solution: Composition per 10.0mL: NaHCO 3 0.2g Preparation of NaHCO 3 Solution: Add NaHCO3 to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. CaCl 2 ·2H 2 O Solution: Composition per 10.0mL: CaCl 2 ·2H 2 O 1.2g Preparation of CaCl 2 ·2H 2 O Solution: Add CaCl 2 ·2H 2 O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: Add components, except NaHCO 3 solution and CaCl 2 ·2H 2 O solution, to distilled/deionized water and bring volume to 980.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Aseptically add 10.0mL of sterile NaHCO 3 solution and 10.0mL of © 2010 by Taylor and Francis Group, LLC . 60.0mL of sterile solution B and 10.0mL of sterile solution C to 1.0L of sterile solution A. Mix thoroughly. Distribute into sterile tubes or flasks. Use: For the cultivation and maintenance of. 4.0g Ethanol, 95% 100.0mL Preparation of Dye Stock Solution, 1000X: Add Bromthymol Blue and Cresol Red to 100.0mL of ethanol. Mix thoroughly. Preparation of Medium: Add components to distilled/deionized water. to form emulsion. Measure 50.0mL of egg yolk emulsion and add to 50.0mL of 0.9% NaCl solution. Mix thoroughly. Filter sterilize. Warm to 45°–50°C. Preparation of Medium: Add components, except