Handbook of Microbiological Media, Fourth Edition part 132 doc

Nutrient Agar No. 2 Diluted 1/100 1305 Plant peptone 5.0g Plant extract 3.0g 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 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 a wide variety of micro- organisms. Nutrient Agar 1.5%, HiVeg with Ascitic Fluid Composition per liter: Agar 15.0g NaCl 8.0g Plant peptone 5.0g Plant extract 3.0g Ascitic fluid 250.0mL pH 7.4 ± 0.2 at 25°C Source: This medium, without ascitic fluid, is available as a premixed powder from HiMedia. Preparation of Medium: Add components, except ascitic fluid, to distilled/deionized water and bring volume to 750.0mL. Mix thorough- ly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to less than 37°C. Aseptically add sterile ascitic fluid. If desired, 0.5g of thallium acetate or 100,000U of penicillin may be added for a more selective medium. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the enrichment of pleuro-pneumonia-like organisms (PPLOs) and Mycoplasma species from clinical specimens. Nutrient Agar with Maltose Composition per liter: Agar 15.0g Maltose 10.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Nocardia corynebacteroi- des. Nutrient Agar No. 2 Composition per liter: Agar 20.0g Meat extract 10.0g Peptone 10.0g NaCl 5.0g pH 7.0 ± 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 a wide variety of bacteria. Nutrient Agar No. 2, Diluted 1/10 (DSMZ Medium 952) Composition per liter: Agar 20.0g Peptone 1.0g Meat extract 1.0g NaCl 0.5g pH 7.1 ± 0.2 at 25°C Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.1. Gently heat and bring to boiling. Distribute into tubes or flasks. Auto- clave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Agromonas oligotrophica. Nutrient Agar No. 2, Diluted 1/10 Composition per liter: Agar 20.0g Meat extract 1.0g Peptone 1.0g NaCl 0.5g pH 7.0 ± 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 Agromonas oligotroph- ica and Kurthia gibsonii. Nutrient Agar No. 2 Diluted 1/100 (DSMZ Medium 952) Composition per liter: Agar 20.0g Peptone 0.1g Meat extract 0.1g NaCl 50.0mg pH 7.1 ± 0.2 at 25°C Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.1. Gently heat and bring to boiling. Distribute into tubes or flasks. Auto- clave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Bacillus pumilus. Nutrient Agar No. 2 Diluted 1/100 Composition per liter: Agar 20.0g Meat extract 0.1g Peptone 0.1g NaCl 50.0mg pH 7.0 ± 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 7.0. © 2010 by Taylor and Francis Group, LLC 1306 Nutrient Agar No. 2, HiVeg Gently heat and bring to boiling. Distribute into tubes or flasks. Auto- clave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Agromonas oligotroph- ica. Nutrient Agar No. 2, HiVeg Composition per liter: Agar 15.0g Plant extract 10.0g Plant peptone 10.0g NaCl 5.0g 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 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 a wide variety of micro- organisms. Nutrient Agar Oxoid CM3 with Phosphate (DSMZ Medium 605a) Composition per liter: Agar 15.0g Peptone 5.0g NaCl 5.0g Na 2 HPO 4 ·12H 2 O 2.39g Yeast extract 2.0g Lab-Lemco beef extract 1.0g KH 2 PO 4 0.45g 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 Devosia riboflavina, Brevun- dimonas diminuta, and Herbaspirillum rubrisubalbicans. Nutrient Agar with 1% Peptone, HiVeg Composition per liter: Agar 15.0g Plant peptone 10.0g Plant extract 5.0g NaCl 5.0g 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 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 a wide variety of micro- organisms. Nutrient Agar, pH 5.0 (Oxoid Nutrient Agar No. 1, pH 5.0) Composition per liter: Agar 15.0g Peptone 5.0g NaCl 5.0g Yeast extract 2.0g Beef extract 1.0g pH 5.0 ± 0.2 at 25°C Source: This medium is available as a premixed powder from Oxoid Unipath. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Adjust pH to 5.0. 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 phenazinium. Nutrient Agar pH 6.0 Composition per liter: Agar 15.0g Peptone 5.0g Beef extract 3.0g pH 6.0 ± 0.2 at 25°C Source: This medium is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Adjust pH to 6.0. 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 microorganisms that prefer a slightly acid nutrient agar. Nutrient Agar pH 6.0 with 0.8% NaCl, HiVeg Composition per liter: Agar 15.0g NaCl 8.0g Plant peptone 5.0g Plant extract 3.0g pH 6.0± 0.2 at 25°C Source: This medium is available as a premixed powder from Hi- Media. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation of microorganisms that prefer a slightly acid nutrient agar. Nutrient Agar pH 6.8 (LMG Medium 2) Composition per liter: Agar 15.0g Peptone 5.0g NaCl 5.0g Na 2 HPO 4 ·12 H 2 O 2.39g Yeast extract 2.0g © 2010 by Taylor and Francis Group, LLC Nutrient Agar pH 9.5 1307 Lab-Lemco beef extract 1.0g KH 2 PO 4 0.45g 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. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure– 121°C. Use: For the cultivation and maintenance of Acidovorax facilis, Pseudo- monas geniculata, Arthrobacter siderocapsulatus, Delftia acidovorans, Pseudomonas spp., and various other heterotrophic bacteria. Nutrient Agar, pH 6.8, HiVeg Composition per liter: Agar 15.0g Plant peptone 5.0g Plant extract 3.0g pH 6.8 ± 0.2 at 25°C Source: This medium is available as a premixed powder 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 cultivation and maintenance of a wide variety of micro- organisms. Nutrient Agar pH 6.8, HiVeg with Blood Composition per liter: Agar 15.0g Plant peptone 5.0g Plant extract 3.0g Sheep blood, defibrinated 50.0mL pH 6.8 ± 0.2 at 25°C Source: This medium, without blood, is available as a premixed pow- der from HiMedia. Preparation of Medium: Add components, except sheep blood, to distilled/deionized water and bring volume to 950.0mL. Mix thorough- ly. Heat with frequent agitation and boil for 1 min to completely dis- solve. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°– 50°C. Aseptically add 50.0mL of sterile, defibrinated sheep blood. Mix thoroughly and pour into sterile Petri dishes. Use: For the cultivation of streptococci and other fastidious microor- ganisms. Nutrient Agar pH 6.8 with Horse Blood (LMG Medium 234) Composition per liter: Agar 15.0g Peptone 5.0g NaCl 5.0g Na 2 HPO 4 ·12 H 2 O 2.39g Yeast extract 2.0g Lab-Lemco beef extract 1.0g KH 2 PO 4 0.45g Horse blood, sterile defibrinated 50.0mL pH 6.8 ± 0.2 at 25°C Preparation of Medium: Add components, except horse blood, to distilled/deionized water and bring volume to 950.0mL. Mix thorough- ly. Adjust pH to 6.8. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add 50.0mL sterile horse blood. Mix thor- oughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of Neisseria weaveri. Nutrient Agar pH 8.0 Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g pH 8.0 ± 0.2 at 25°C . Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Adjust pH to 8.0. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Allow tubes to cool in a slanted position. Use: For the cultivation and maintenance of Bacillus alcalophilus. Nutrient Agar pH 9.0 (ATCC Medium 2029) Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g pH 9.0 ± 0.1 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. Adjust pH to 9.0. 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 alkilophilic bacteria. Nutrient Agar pH 9.5 Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g Na 2 CO 3 solution variable pH 9.5–10.0 at 25°C Na 2 CO 3 Solution: Composition per 100.0mL: Na 2 CO 3 5.0g Preparation of Na 2 CO 3 Solution: Add Na 2 CO 3 to distilled/de- ionized water and bring volume to 100.0mL. Mix thoroughly. Auto- clave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Preparation of Medium: Add components, except Na 2 CO 3 solu- tion, to distilled/deionized water and bring volume to 1.0L. Mix thor- oughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Adjust pH to 9.5 with sterile Na 2 CO 3 solution. Mix thoroughly. Pour into sterile Petri dishes or dis- tribute into sterile tubes. Allow tubes to cool in a slanted position. Use: For the cultivation and maintenance of Bacillus alcalophilus. © 2010 by Taylor and Francis Group, LLC 1308 Nutrient Agar pH 10.0 Nutrient Agar pH 10.0 (ATCC Medium 2030) Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g pH 10.0 ± 0.1 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. Adjust pH to 10.0. 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 alkilophilic bacteria. Nutrient Agar with Cysteine (ATCC Medium 812) Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g L-Cysteine 0.1g pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Salmonella choleraesuis. Nutrient Agar with Dihydrostreptomycin Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g Dihydrostreptomycin solution 10.0mL pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Dihydrostreptomycin Solution: Composition per 10.0mL: Dihydrostreptomycin 0.625g Preparation of Dihydrostreptomycin Solution: Add dihy- drostreptomycin to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except dihydrostrepto- mycin solution, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add sterile dihydrostreptomycin solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of Escherichia coli, Micro- coccus luteus, Shigella species, and Vibrio cholerae. Nutrient Agar with Erythromycin Composition per liter: Agar 15.0g Peptone 5.0g Beef extract 3.0g Erythromycin solution 10.0mL pH 6.8 ± 0.2 at 25°C. Erythromycin Solution: Composition per 10.0mL: Erythromycin 0.6g Preparation of Erythromycin Solution: Add erythromycin to distilled/deionized water and bring volume to 10.0mL. Mix thorough- ly. Filter sterilize. Preparation of Medium: Add components, except erythromycin solution, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add sterile erythromycin solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of Micrococcus luteus. Nutrient Agar with Ethanolamine Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g Ethanolamine 2.0mL pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Flavobacterium tirreni- cum and Pseudomonas species. Nutrient Agar with Ethylene Glycol Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g Ethylene glycol 2.0mL pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Pseudomonas putida. Nutrient Agar with Glucose Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g © 2010 by Taylor and Francis Group, LLC Nutrient Agar with 1% Methanol 1309 Beef extract 3.0g Glucose 10.0g pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Amycolata saturnea, Arthrobacter species, Corynebacterium species, Curtobacterium flaccum- faciens, Deinococcus radiodurans, Escherichia coli, Hafnia alvei, Micro- coccus aurantiacus, Myxomicrobium multiplex, Nocardia petroleophila, Nocardia species, Pseudomonas species, Rhodococcus rhodochrous, Streptomyces piedadensis, and Xanthomonas species. Nutrient Agar with Horse Serum (LMG Medium 38) Composition per liter: Agar 15.0g Peptone 5.0g NaCl 5.0g Yeast extract 2.0g Lab-Lemco beef extract 1.0g Horse serum 100.0mL pH 7.4 ± 0.2 at 25°C Preparation of Medium: Add components, except horse serum, 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 pres- sure–121°C. Cool medium to 45–50°C. Aseptically add 100.0mL of sterile horse serum. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation of fastidious bacteria. Nutrient Agar with Horse Serum Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g Horse serum 100.0mL pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components, except horse serum, to distilled/deionized water and bring volume to 900.0mL. Mix thorough- ly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add sterile horse se- rum. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of Alysiella filiformis and Simonsiella crassa. Nutrient Agar with 10% Horse Serum (DSMZ Medium 302) Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g Horse serum 100.0mL pH 7.0 ± 0.2 at 25°C Preparation of Medium: Add components, except horse serum, to distilled/deionized water and bring volume to 900.0mL. Mix thorough- ly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Aseptically add 100.0mL filter steril- ized horse serum. Mix thoroughly. Pour into sterile Petri dishes or aseptically distrubute into tubes. Use: For the isolation and cultivation of Simonsiella crassa. Nutrient Agar with Manganese (NAMn) Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g Manganese sulfate solution 1.0mL pH 6.8 ± 0.2 at 25°C Manganese Sulfate Solution: Composition per 100.0mL: MnSO 4 ·H 2 O 3.08g Preparation of Manganese Sulfate Solution: Add MnSO 4 ·H 2 O to distilled/deionized water and bring volume to 100.0mL. Mix thor- oughly. 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 of Bacillus species from canned foods. For enhanced spore production by Bacillus species. Nutrient Agar with Manganese Sulfate (ATCC Medium 1859) Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g Manganese sulfate solution 1.0mL pH 6.8 ± 0.2 at 25°C Manganese Sulfate Solution: Composition per 100.0mL: MnSO 4 ·H 2 O 0.5g Preparation of Manganese Sulfate Solution: Add MnSO 4 ·H 2 O to distilled/deionized water and bring volume to 100.0mL. Mix thor- oughly. 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 cultivation of Paenibacillus chondroitinus and Paenibacillus alginolyticus. Nutrient Agar with 1% Methanol (ATCC Medium 620) Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g © 2010 by Taylor and Francis Group, LLC 1310 Nutrient Agar with 2% Methanol Beef extract 3.0g Methanol 10.0mL pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Filter sterilize methanol. Add compo- nents, except methanol, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Gently heat and bring to boiling. Auto- clave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Asepti- cally add sterile methanol. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of Bacillus species, Methyl- omonas clara, and Pseudomonas methanolica. Nutrient Agar with 2% Methanol (ATCC Medium 628) Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g Methanol 20.0mL pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Filter sterilize methanol. Add compo- nents, except methanol, to distilled/deionized water and bring volume to 980.0mL. Mix thoroughly. Gently heat and bring to boiling. Auto- clave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Asepti- cally add sterile methanol. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of Pseudomonas species. Nutrient Agar with 0.5% Sodium Chloride Composition per liter: Agar 15.0g NaCl 5.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Agrobacterium tumefa- ciens, Escherichia coli, Pseudomonas aeruginosa, Salmonella choler- aesuis, Shigella dysenteriae, Shigella flexneri, Vibrio species, and Yersinia species. Nutrient Agar with 0.5% Sodium Chloride, pH 9.5-10.0 (LMG Medium 253) Composition per liter: Agar 15.0g NaCl 10.0g Peptone 5.0g Yeast extract 2.0g Lab-Lemco beef extract 1.0g pH 9.5–10.0 Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°-50°C. Adjust pH to 9.5-10.0 with ster- ile Na 2 CO 3 solution. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Bacillus spp. Nutrient Agar with 1.5% Sodium Chloride Composition per liter: NaCl 15.0g Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Photobacterium leiog- nathi, Pseudomonas fluorescens, and Vibrio natriegens. Nutrient Agar with 2% Sodium Chloride Composition per liter: NaCl 20.0g Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g 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 Vibrio alginolyticus, Vibrio bivalvii, Vibrio mediterranei, Vibrio natriegens, Vibrio ordali, Vibrio orientalis, Vibrio parahaemolyticus, Vibrio pelagigius, and Vibrio vulnificus. Nutrient Agar with 3% Sodium Chloride (LMG Medium 257) Composition per liter: NaCl 35.0g Agar 15.0g Peptone 5.0g Yeast extract 2.0g Lab-Lemco beef extract 1.0g 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 and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Adjust pH to 9.5–10.0 with sterile Na 2 CO 3 solution. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Bacillus cirroflagellosus. © 2010 by Taylor and Francis Group, LLC Nutrient Agar with Nalidixic Acid 1311 Nutrient Agar with 3% Sodium Chloride Composition per liter: NaCl 30.0g Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Bacillus species, Altero- monas nigrifaciens, Halococcus species, Planococcus citreus, Pseudomonas beijerinckii, Staphylococcus species, Streptococcus pyogenes, and Vibrio species. Nutrient Agar with 5% Sodium Chloride, pH 9.5–10.0 (LMG Medium 254) Composition per liter: NaCl 55.0g Agar 15.0g Peptone 5.0g Yeast extract 2.0g Lab-Lemco beef extract 1.0g pH 9.5–10.0 Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Adjust pH to 9.5–10.0 with sterile Na 2 CO 3 solution. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Bacillus haloalkaliphilus. Nutrient Agar with 10% Sodium Chloride Composition per liter: NaCl 100.0g Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Paracoccus halodenitrif- icans and Micrococcus species. Nutrient Agar with 10% Sodium Chloride and Maltose Composition per liter: NaCl 100.0g Agar 15.0g Maltose 10.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Paracoccus halodenitrif- icans and Micrococcus species. Nutrient Agar with 0.5% Sodium Chloride and Sodium Citrate Composition per liter: Agar 15.0g NaCl 5.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g Sodium citrate 2.94g pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Escherichia coli. Nutrient Agar with Nalidixic Acid (ATCC Medium 2089) Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g NaCl 5.0g Beef extract 3.0g Nalidixic acid solution 10.0mL pH 6.8 ± 0.2 at 25°C Nalidixic Acid Solution: Composition per 10.0mL: Nalidixic acid 0.1g Preparation of Nalidixic Acid Solution: Add nalidixic acid to distilled/deionized water and bring volume to 10.0mL. Mix thorough- ly. Filter sterilize. Preparation of Medium: Add components, except nalidixic acid solution, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Gently heat while stirring and bring to boiling. Auto- clave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Asepti- cally add sterile nalidixic acid solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation of nalidixic acid-resistant bacteria. Nutrient Agar with Phosphate See: Nutrient Agar, Buffered © 2010 by Taylor and Francis Group, LLC 1312 Nutrient Agar with Phytone™ Nutrient Agar with Phytone™ Composition per liter: Agar 15.0g Phytone™ 10.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g 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 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 a wide variety of bacteria. Nutrient Agar with Potato Starch Composition per liter: Agar 15.0g Potato starch 10.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g pH 6.8 ± 0.2 at 25°C Source: Nutrient agar 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 Bacillus polymyxa and Bacillus subtilis. Nutrient Agar with Soil Extract Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g Soil extract 250.0mL pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Soil Extract: Composition per 300.0mL: African Violet soil 115.5g Na 2 CO 3 0.3g Preparation of Soil Extract: Add components to tap water and bring volume to 300.0mL. Autoclave for 60 min at 15 psi pressure– 121°C. Filter through Whatman filter paper. 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. Use: For the cultivation and maintenance of Auerobacterium species, Bacillus species, and Saccharomonospora viridis. Nutrient Agar with Streptomycin Composition per liter: Agar 15.0g Peptone 5.0g Beef extract 3.0g Streptomycin solution 10.0mL pH 6.8 ± 0.2 at 25°C Streptomycin Solution: Composition per 10.0mL: Streptomycin 0.02g Preparation of Streptomycin Solution: Add streptomycin to dis- tilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except streptomycin solution, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add sterile streptomycin solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of Corynebacterium glutam- icum and Corynebacterium herculis. Nutrient Agar with Sucrose Composition per liter: Sucrose 20.0g Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Pseudomonas species. Nutrient Agar with 2% Sucrose (ATCC Medium 1297) Composition per liter: Sucrose 20.0 g Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g 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 of osmophilic bacteria Nutrient Agar with Tetracycline Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g Tetracycline solution 10.0mL pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. © 2010 by Taylor and Francis Group, LLC Nutrient Broth 1313 Tetracycline Solution: Composition per 10.0mL: Tetracycline 0.025g Preparation of Tetracycline Solution: Add tetracycline to dis- tilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except tetracycline so- lution, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add sterile tet- racycline solution. Mix thoroughly. Pour into sterile Petri dishes or dis- tribute into sterile tubes. Use: For the cultivation and maintenance of Salmonella choleraesuis. Nutrient Agar with Uracil Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g Uracil 0.01g pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Escherichia coli. Nutrient Agar with 5% Urea Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g Urea solution 100.0mL pH 6.8 ± 0.2 at 25°C Urea Solution: Composition per 100.0mL: Urea 50.0g Preparation of Urea Solution: Add urea to distilled/deionized wa- ter and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except urea solution, to distilled/deionized water and bring volume to 900.0mL. Mix thorough- ly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add 100.0mL of sterile urea solution. Mix thoroughly. Pour into sterile Petri dishes or asepti- cally distribute into sterile tubes. Use: For the cultivation and maintenance of Aspergillus species. Nutrient Agar with Ureum (LMG Medium 139) Composition per liter: Ureum 20.0g Agar 15.0g Peptone 5.0g NaCl 5.0g Yeast extract 2.0g Lab-Lemco beef extract 1.0g 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 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 Sporosarcina pasteurii. Nutrient Agar with V-8™ Juice Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g V-8™ juice 200.0mL pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Pseudomonas tolaasii. Nutrient Agar with Yeast Extract Composition per liter: Yeast extract 20.0g Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g pH 6.8 ± 0.2 at 25°C Source: Nutrient agar is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Bacillus anthracis and Comamonas testosteroni. Nutrient Broth Composition per liter: Peptone 5.0g NaCl 5.0g Yeast extract 2.0g Beef extract 1.0g pH 7.4 ± 0.2 at 25°C Source: This medium is available as a premixed powder from Oxoid Unipath. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of a wide variety of nonfastidious microorgan- isms. © 2010 by Taylor and Francis Group, LLC 1314 Nutrient Broth Nutrient Broth (DSMZ Medium 948) Composition per liter: Peptone 5.0g NaCl 5.0g Yeast extract 2.0g Lab-Lemco beef extract 1.0g 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. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of Psychrobacter urativorans. Nutrient Broth Composition per liter: Pancreatic digest of gelatin 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. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of a wide variety of nonfastidious microorgan- isms. Nutrient Broth with Bovine Serum Composition per liter: NaCl 10.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g Bovine serum 100.0mL pH 6.9 ± 0.2 at 25°C Preparation of Medium: Add components, except bovine serum, to distilled/deionized water and bring volume to 900.0mL. Mix thor- oughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°– 50°C. Aseptically add sterile bovine serum. Mix thoroughly. Aseptical- ly distribute into sterile tubes or flasks. Use: For the cultivation and maintenance of Pseudomonas anguil- liseptica. Nutrient Broth, Diluted 1:100 Nutrient Broth: Composition per liter: Pancreatic digest of gelatin 5.0g Beef extract 3.0g pH 6.8 ± 0.2 at 25°C Preparation of Nutrient Broth: Add components to distilled/de- ionized water and bring volume to 1.0L. Mix thoroughly. Preparation of Medium: Add 10.0mL of nutrient broth to distilled/ deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation and maintenance of Agromonas oligotroph- ica. Nutrient Broth Glycerol Medium Composition per liter: Glycerol 100.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g pH 7.2–7.4 ± 0.2 at 25°C Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of Pseudomonas species. Nutrient Broth with Manganese Sulfate Composition per liter: MnSO 4 5.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g pH 6.9 ± 0.2 at 25°C Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of Bacillus chondroitinus and Bacillus algi- nolyticus. Nutrient Broth with Rifampicin Composition per liter: Peptone 5.0g NaCl 5.0g Yeast extract 2.0g Beef extract 1.0g Rifampicin solution 10.0mL pH 7.4 ± 0.2 at 25°C Rifampicin Solution: Composition per 10.0mL: Rifampicin 10.0mg Preparation of Rifampicin Solution: Add rifampicin to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except rifampicin solu- tion, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Adjust pH to 7.4. Autoclave for 15 min at 15 psi pressure– 121°C. Cool to room temperature. Aseptically add 10.0mL of sterile ri- fampicin solution. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation of Agrobacterium tumefaciens. Nutrient Broth with 6% Sodium Chloride Composition per liter: NaCl 60.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g 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. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of organisms in water, sewage, feces, and other materials. For the cultivation and maintenance of Paracoccus halodenitrificans. © 2010 by Taylor and Francis Group, LLC . 0.5g of thallium acetate or 100,000U of penicillin may be added for a more selective medium. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the enrichment of pleuro-pneumonia-like. and maintenance of Neisseria weaveri. Nutrient Agar pH 8.0 Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g pH 8.0 ± 0.2 at 25°C . Preparation of Medium: Add. tubes. Use: For the cultivation of a wide variety of bacteria. Nutrient Agar with Potato Starch Composition per liter: Agar 15.0g Potato starch 10.0g Pancreatic digest of gelatin 5.0g Beef extract