Handbook of Microbiological Media, Fourth Edition part 101 docx

Magnetospirillum Medium 995 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 Yersinia enterocolitica from foods. Magnetic Spirillum Growth Medium, Revised (MSGM, Revised) Composition per liter: Agar 1.3g KH 2 PO 4 0.68g Tartaric acid 0.37g Succinic acid 0.37g NaNO 3 0.12g Sodium acetate 0.05g Ascorbic acid 0.035g Wolfe’s vitamin solution 10.0mL Wolfe’s mineral solution 5.0mL Ferric quinate solution 2.0mL Resazurin (0.1% solution) 0.45mL pH 6.75 ± 0.2 at 25°C Wolfe’s Vitamin Solution: Composition per liter: Pyridoxine·HCl 10.0mg Thiamine·HCl 5.0mg Riboflavin 5.0mg Nicotinic acid 5.0mg Calcium pantothenate 5.0mg p-Aminobenzoic acid 5.0mg Thioctic acid 5.0mg Biotin 2.0mg Folic acid 2.0mg Cyanocobalamin 100.0μg Preparation of Wolfe’s Vitamin Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Wolfe’s Mineral Solution: Composition per liter: MgSO 4 ·7H 2 O 3.0g Nitriloacetic acid 1.5g NaCl 1.0g MnSO 4 ·H 2 O 0.5g FeSO 4 ·7H 2 O 0.1g CoCl 2 ·6H 2 O 0.1g CaCl 2 0.1g ZnSO 4 ·7H 2 O 0.1g CuSO 4 ·5H 2 O 0.01g AlK(SO 4 ) 2 ·12H 2 O 0.01g H 3 BO 3 0.01g Na 2 MoO 4 ·2H 2 O 0.01g Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH to 6.5 with KOH. Add remaining components. Add distilled/deionized water to 1.0L. Ferric Quinate Solution: Composition per 100.0mL: FeCl 3 0.27g Quinic acid 0.19g Preparation of Ferric Quinate Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: To 1.0L of distilled/deionized water add components in the following order: Wolfe’s vitamin solution, Wolfe’s min- eral solution, ferric quinate solution, resazurin, KH 2 PO 4 , NaNO 3 , ascorbic acid, tartaric acid, succinic acid, sodium acetate, and agar. Mix thoroughly after each addition. Adjust pH to 6.75 with NaOH. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically distribute into sterile screw-capped tubes. Fill tubes to capacity with medium. Use a heavy inoculum in each tube and do not introduce a headspace of air. Screw down caps tightly. Use: For the cultivation and maintenance of Aquaspirillum magnetot- acticum. Magnetospirillum Medium (DSMZ Medium 380) Composition per liter: KH 2 PO 4 0.68g L(+)-Tartaric acid 0.37g Succinic acid 0.37g NaNO 3 0.12g Na-thioglycolate 0.05g Na-acetate 0.05g Resazurin 0.5mg Vitamin solution 10.0mL Trace elements solution 5.0mL Ferric quinate solution 2.0mL pH 6.8 ± 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. 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 © 2010 by Taylor and Francis Group, LLC 996 Magnetospirillum 2 Medium 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. Ferric Quinate Solution: Composition per 100.0mL: FeCl 3 ·6H 2 O 0.45g Quinic acid 0.19g Preparation of Ferric Quinate Solution: Add components to distilled/deionized water and bring volume to 100.0mL. Sparge with N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Preparation of Medium: Add components, except vitamin solu- tion and ferric quinate solution, to distilled/deionized water and bring volume to 988.0mL. Purge medium with N 2 gas for 10 min. Mix thor- oughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Aseptically and anerobically add 10.0mL vitamin solution and 2.0mL ferric quinate solution. Mix thoroughly. Purge medium with N 2 gas for 10 min. Under the same atmosphere, anaerobically fill tubes to 1/3 of their volume and seal. Autoclave at 121°C for 15 min. Before inocula- tion, add sterile air (with hypodermic syringe through the rubber clo- sure) to 1% O 2 concentration in the gas phase. Use: For the cultivation of Magnetospirillum magnetotacti- cum=Aquaspirillum magnetotacticum, and Magnetospirillum gryph- iswaldense. Magnetospirillum 2 Medium Composition per liter: Sodium acetate 1.0g K 2 HPO 4 0.5g Sodium thioglycolate 0.5g NH 4 Cl 0.1g Yeast extract 0.1g Ferric citrate 20.0μg 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 screw- capped tubes in 5.0mL volumes. Autoclave for 15 min at 15 psi pres- sure–121°C. Allow medium to stand upright at room temperature for 2 to 3 days before inoculation. Do not shake. Use: For the cultivation and maintenance of Magnetospirillum gryph- iswaldense. Magnetospirillum Semi-solid Medium (DSMZ Medium 380) Composition per liter: Agar 1.3g KH 2 PO 4 0.68g L(+)-Tartaric acid 0.37g Succinic acid 0.37g NaNO 3 0.12g Na-thioglycolate 0.05g Na-acetate 0.05g Resazurin 0.5mg Vitamin solution 10.0mL Trace elements solution 5.0mL Ferric quinate solution 2.0mL pH 6.8 ± 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. 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. Ferric Quinate Solution: Composition per 100.0mL: FeCl 3 ·6H 2 O 0.45g Quinic acid 0.19g Preparation of Ferric Quinate Solution: Add components to distilled/deionized water and bring volume to 100.0mL. Sparge with N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°C. Preparation of Medium: Add components, except vitamin solu- tion and ferric quinate solution, to distilled/deionized water and bring volume to 988.0mL. Purge medium with N 2 gas for 10 min. Mix thor- oughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°C. Aseptically and anerobically add 10.0mL vitamin solution and 2.0mL ferric quinate solution. Mix thoroughly. Purge medium with N 2 gas for 10 min. Dispense 12mL of medium per 16 x 150mm anaerobe screw- cap tube under N 2 gas. Prior to inoculation, remove caps briefly under air, tighten the caps again, and wait several hours to establish oxygen gradients. The medium should be slightly pink in color. Strongly re- duced conditions will not support growth of the organism. During growth O 2 will be consumed, resazurin decolorized, and the pH in- creased. Feed oxygen (by adding air) and succinic acid from sterile 0.05M solution (to maintain pH below 7.0). If higher densities of mag- netic cell are wanted, ferric quinate also can be fed. For transfer use cell material which has been concentrated at the glass wall of the culture vessel by means of a magnetic rod attached outside. © 2010 by Taylor and Francis Group, LLC Maleate Medium for Pseudomonas fluorescens with Glucose and Phenol Red 997 Use: For the cultivation of Magnetospirillum magnetotacticum (Aquaspirillum magnetotacticum) and Magnetospirillum gryphiswald- ense. Maintenance HiVeg Medium for B. subtilis ATCC 6633 Composition per liter: Agar 15.0g Plant peptone 6.0g Plant hydrolysate 4.0g Yeast extract 3.0g Plant extract 1.5g Glucose 1.0g pH 7.2 ± 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 until boiling. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation of Bacillus subtilis. Maintenance of L Antigen in Neisseria Composition per liter: Proteose peptone No. 3 20.0g Agar 15.0g Na 2 HPO 4 5.0g NaCl 5.0g Glucose 0.5g Rabbit blood, defibrinated 100.0mL pH 7.4–7.6 at 25°C Preparation of Medium: Add components, except rabbit blood, to distilled/deionized water and bring volume to 900.0mL. Mix thorough- ly. Gently heat while stirring until boiling. Autoclave for 20 min at 15 psi pressure–121°C. Cool to 60°C. Aseptically add 100.0mL of sterile, defibrinated rabbit blood. Maintain at 75°C while shaking for 30 min. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of Neisseria gonorrhoeae. Maintenance SCY Medium See: SCY Medium Maintenance (SCY) HiVeg Medium Composition per liter: Agar 10.0g Sucrose 1.0g Plant hydrolysate 0.91g Yeast extract 0.25g NaCl 0.05g Papaic digest of soybean meal 0.03g K 2 HPO 4 0.02g Thiamine 0.4mg pH 7.3 ± 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. Filter sterilize. Use: For the cultivation and maintenance of iron and sulfur bacteria. Malachite Green Broth Composition per liter: Peptone 15.0g Beef extract 9.0g Malachite Green 0.01mg pH 7.3 ± 0.2 at 25°C Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of Pseudomonas aeruginosa. Maleate Medium for Pseudomonas fluorescens Composition per liter: Agar 15.0g K 2 HPO 4 1.13g NH 4 NO 3 1.0g KH 2 PO 4 0.48g MgSO 4 ·7H 2 O 0.2g Potassium maleate solution 8.61mL pH 7.0 ± 0.2 at 25°C Potassium Maleate Solution: Composition per liter: Maleic acid 116.07g KOH (10N solution) 200.0mL Preparation of Potassium Maleate Solution: Add maleic acid to distilled/deionized water and bring volume to 600.0mL. Slowly add KOH solution (generates heat). Bring volume to 1.0L with distilled/de- ionized water. Adjust pH to 7.0. Filter sterilize. Preparation of Medium: Add components, except potassium maleate solution, to distilled/deionized water and bring volume to 991.4mL. Mix thoroughly. Gently heat while stirring until boiling. Au- toclave for 20 min at 15 psi pressure–121°C. Cool to 50°C. Aseptically add 8.61mL of the potassium maleate solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of Pseudomonas fluorescens and Mycoplasma pneumoniae. Maleate Medium for Pseudomonas fluorescens with Glucose and Phenol Red Composition per liter: Agar 15.0g Glucose 10.0g K 2 HPO 4 1.13g NH 4 NO 3 1.0g KH 2 PO 4 0.48g MgSO 4 ·7H 2 O 0.2g Phenol Red 0.04g Potassium maleate solution 8.61mL pH 7.0 ± 0.2 at 25°C Potassium Maleate Solution: Composition per liter: Maleic acid 116.07g KOH (10N solution) 200.0mL Preparation of Potassium Maleate Solution: Add maleic acid to distilled/deionized water and bring volume to 600.0mL. Slowly add KOH solution (generates heat). Bring volume to 1.0L with distilled/de- ionized water. Adjust pH to 7.0. Filter sterilize. © 2010 by Taylor and Francis Group, LLC 998 Malonate Broth Preparation of Medium: Add components, except potassium maleate solution, to distilled/deionized water and bring volume to 991.4mL. Mix thoroughly. Gently heat while stirring until boiling. Au- toclave for 20 min at 15 psi pressure–121°C. Cool to 50°C. Aseptically add 8.61mL of the potassium maleate solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of Pseudomonas fluorescens and Mycoplasma pneumoniae. Malonate Broth Composition per liter: Sodium malonate 3.0g NaCl 2.0g (NH 4 ) 2 SO 4 2.0g K 2 HPO 4 0.6g KH 2 PO 4 0.4g Bromthymol Blue 0.025g pH 6.7 ± 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. Avoid intro- duction of carbon and nitrogen from other sources. Use: For the cultivation and differentiation of coliforms and other enteric organisms, particularly Enterobacter and Escherichia, based on their ability to utilize malonate as the sole carbon source and ammo- nium sulfate as the sole nitrogen source. Malonate-utilizing organisms turn the medium blue. Malonate Broth, Ewing Modified Composition per liter: Sodium malonate 3.0g NaCl 2.0g (NH 4 ) 2 SO 4 2.0g Yeast extract 1.0g Glucose 0.25g K 2 HPO 4 0.6g KH 2 PO 4 0.4g Bromthymol Blue 0.025g pH 6.7 ± 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 and differentiation of coliforms and other enteric organisms, particularly Enterobacter and Escherichia, based on their ability to utilize malonate as a carbon source and ammonium sul- fate as a nitrogen source. The small amount of yeast extract and glu- cose encourages the growth of some organisms that may be distressed or fail to respond. Malonate-utilizing organisms turn the medium blue. Malt Agar Composition per liter: Malt extract 30.0g Agar 15.0g pH 5.5 ± 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 until boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–118°C. Do not overheat or agar will not harden. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation of yeasts and molds. Malt Agar Composition per liter: Agar 20.0g Malt extract 12.5g Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat until 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 fungi. Malt Agar, 1/3 Strength (ATCC Medium 2365) Composition per liter: Agar 15.0g Malt extract 10.0g pH 5.5 ± 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 until boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–118°C. Do not overheat or agar will not harden. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation of yeasts and molds. Malt Agar, Blakeslee Composition per liter: Glucose 20.0g Malt extract 20.0g Agar 16.0g Mycological peptone 1.0g 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 Acremonium chrysogenum, Agaricus bisporus, Agrocybe aegerita, Armillaria mellea, Aspergillus spe- cies, Aureobasidium pullulans, Basidiomycetes species, Candida species, Ceratocystis adiposa, Citeromyces matritensis, Cladosporium cucumeri- num, Cochliobolus miyaheanus, Colletotrichum destructivum, Colletotri- chum lindemuthianum, Cryptococcus albidus, numerous other Cryptococ- cus species, Debaryomyces polymorphus, Diaporthe magnusii, Diaporth- ephaseolorum, Drechslera spicifera, Fusarium graminearum, Geotri- chum lactis, other Geotrichum species, Hanseniaspora uvarum, Hanse- niaspora valbyensis, Hansenula subpelliculosa, Humicola species, Kloeckera apiculata, Kloeckera corticis, Kluyveromyces lodderi, Kluyver- omyces marxianus, Metschnikowia pulcherrima, Monilinia fructigena, © 2010 by Taylor and Francis Group, LLC Malt Agar 4% with Lupine Stems 999 Myrothecium verrucaria, Myxozyma melibiosi, Nadsonia fulvescens, Neu- rospora crassa, Octosporomyces octosporus, Pachysolen tannophilus, Paecilomyces variotii, Penicillium aurantiogriseum, many other Penicil- lium species, Pithoascus schumacheri, Rhizoctonia crocorum, Rhizomu- cor miehei, Rhodosporidium toruloides, Rhodotorula glutinis, Rhodoto- rula minuta, Rhodotorula rubra, Saccharomyces cerevisiae, Saccharomy- ces exiguus, Saccharomyces ludwigii, Saccharomyces capsularis, Saccha- romyces fibuligera, Schizosaccharomyces pombe, Sporobolomyces para- roseus, Sporobolomyces salmonicolor, Sporopachydermia lactativora, Stachybotrys species, Talaromyces emersonii, Taphrina deformans, Toru- laspora delbrueckii, Trichoderma harzianum, Trichoderma koningii, Trichoderma longibrachiatum, Trichoderma reesei, Trichoderma viride, Trichosporon beigelii, Wickerhamia fluorescens, Yarrowia lipolytica, and Zygosaccharomyces veronae. Malt Agar 1% (MA1) Composition per liter: Agar 15.0g Malt extract 10.0g 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 of Acrodontium griseum, Acrodontium simplex, Acrophialophora fusispora, Acrothecium tenebrosum, Acrothecium capsic, Agaricus bisporus, Chaetocladium jonesii, Chaetomium globo- sum, Chaetomium cupreum, Chaetomium irregulare, Farlowiella carmichaeliana, and many other filamentous fungi. Malt Agar 2% (MA2) Composition per liter: Malt extract 20.0g Agar 15.0g 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 of Acrophialophora fusispora, Agaricus augustus, Ceratocystis penicillata, Chaetocladium brefeldii, and many other fungi. Malt Agar 4% (MA4) Composition per liter: Malt extract 40.0g Agar 15.0g 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 of Chaetomium globosum, Chaetomium indi- cum, Chaetomium funicola, Chaetomium megalocarpum, Chaetomium murorum, Chaetomium seminudum, Chaetomium pachypodioides, Chaetomium perlucidum, Chaetomium quadrangulatum, Chaetomium reflexum, Chaetomium subaffine, Chaetomium subspirilliferum, Chaeto- mium succineum, Chaetomium luknowense, Daedalea quercina, Mycosphaerella ligulicola, Polypaecilum pisce, and many other fungi. Malt Agar 8% (MA8) Composition per liter: Malt extract 80.0g Agar 15.0g 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 of Moniliella pollinis. Malt Agar with 0.5% CaCO 3 Composition per liter: Malt extract 20.0g Glucose 20.0g Agar 15.0g CaCO 3 5.0g Mycological peptone 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. 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 Brettanomyces anomalus, Brettanomyces bruxellensis, Brettanomyces claussenii, Brettanomyces lambicus, Dekkera bruxellensis, and Dekkera intermedia. Malt Agar with 2% Malt Composition per liter: Agar 20.0g Malt extract 20.0g pH 6.5 ± 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 6.5. 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 a variety of yeasts and other fungi. Malt Agar 2% with Lupine Stems (MA2 with Lupine Stems) Composition per liter: Malt extract 20.0g Agar 15.0g Lupine stems variable Preparation of Medium: Add components, except lupine stems, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute 6.0mL volumes into tubes. Cut lupine stems into 8.0cm-long pieces. Add 2–3 lupine stems per tube. Autoclave for 15 min at 15 psi pressure–121°C. Allow tubes to cool in a slanted position. Use: For the cultivation of Chaetomium bostrychodes. Malt Agar 4% with Lupine Stems (MA4 with Lupine Stems) Composition per liter: Malt extract 40.0g Agar 15.0g Lupine stems variable © 2010 by Taylor and Francis Group, LLC 1000 Malt Dextrose Peptone Agar Preparation of Medium: Add components, except lupine stems, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute 6.0mL volumes into tubes. Cut lupine stems into 8.0cm-long pieces. Add 2–3 lupine stems per tube. Autoclave for 15 min at 15 psi pressure–121°C. Allow tubes to cool in a slanted position. Use: For the cultivation of Chaetomium indicum and Chaetospermum chaetosporum. Malt Dextrose Peptone Agar (MDPA) Composition per liter: Agar 25.0g Malt extract 20.0g Glucose 20.0g Peptone 1.0g 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 of Aspergillus aeneus, Aspergillus candidus, Odontia uda, Oidiodendron chlamydosporicum, Oidiodendron cerealis, Oidiodendron flavum, Oidiodendron griseum, Oidiodendron perico- nioides, Oidiodendron tenuissimum, Penicillium spinulosum, and other fungi. Malt Dextrose 40% Peptone Agar (MDPA 40) Composition per liter: Glucose 400.0g Agar 25.0g Malt extract 20.0g Peptone 1.0g 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 of Aspergillus penicilloides. Malt 4% Dextrose Peptone Yeast Agar (MDPYA4) Composition per liter: Malt extract 40.0g Agar 20.0g Glucose 20.0g Peptone 1.0g Yeast extract 1.0g 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 of Candida glabrata, Candida haemulonii, Can- dida lactis-condensi, Candida magnoliae, Candida nemodendra, Met- schnikowia pulcherrima, Metschnikowia reukaufii, Phoma glomerata, Saccharomyces exiguus, and Trigonopsis variabilis. Malt 4% Dextrose Yeast Agar (MDYA4) Composition per liter: Malt extract 40.0g Agar 20.0g Glucose 20.0g Yeast extract 1.0g 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 of Torulaspora delbrueckii. Malt Extract Agar Composition per liter: Malt extract 30.0g Agar 15.0g Peptone 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 20 min at 10 psi pressure–115°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation of Xanthomonas species. Malt Extract Agar (MEA) Composition per liter: Agar 20.0g Glucose 20.0g Malt extract 20.0g Peptone 1.0g pH 5.5 ± 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 isolation, cultivation, and identification of heat-resistant filamentous fungi (molds) from foods. Malt Extract Agar Composition per liter: Malt extract 30.0g Agar 15.0g Mycological peptone 5.0g pH 5.4 ± 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. Gently heat while stirring until boiling. Distribute into tubes or flasks. Autoclave for 10 min at 15 psi pressure–115°C. Do not overheat or agar will not harden. If a lower pH (3.5) is desired, cool medium to 55°C and aseptically add 100.0mL of sterile lactic acid. Pour into sterile Petri dishes or distribute into sterile tubes. © 2010 by Taylor and Francis Group, LLC Malt Extract Agar, Half Strength 1001 Use: For the detection, isolation, and enumeration of yeasts and molds. The addition of lactic acid suppresses bacterial growth. Malt Extract Agar Composition per liter: Malt extract 30.0g Agar 20.0g Chlortetracycline solution 10.0mL pH 5.5 ± 0.2 at 25°C Chlortetracycline Solution: Composition per 10.0mL: Chlortetracycline 0.04mg Preparation of Chlortetracycline Solution: Add chlortetracy- cline to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except chlortetracy- cline 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 chlortetracycline solution. Mix thoroughly. Pour into sterile Petri dishes in 20.0mL volumes. Use: For the cultivation of yeasts and filamentous fungi (molds) from cosmetics. Malt Extract Agar Composition per liter: Malt extract 20.0g Agar 15.0g Peptone 5.0g 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 Candida melibiosica, Cryptococcus curvatus, Kluyveromyces species, Metschnikowia hawaiiensis, Rhodosporidium paludigenum, Saccharomyces cerevi- siae, Saccharomycodes ludwigii, Stephanoascus species, and Trichos- poron nigrescens. Malt Extract Agar (ATCC Medium 109) Composition per liter: Agar 15.0g Maltose 12.75g Dextrin 2.75g Glycerol 2.35g Pancreatic digest of gelatin 0.78g pH 4.6 ± 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 until boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–118°C. Do not overheat or agar will not harden. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of yeasts, molds, and Fla- vobacterium lucecoloratum. Malt Extract Agar (MEA) Composition per liter: Glucose 20.0g Malt extract 20.0g Agar 15.0g Peptone 1.0g 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 of Arthrinium phaeospermum, Aspergillus fumigatus, Aspergillus clavatus, Penicillium verruculosum, and Peni- cillium spinulosum. Malt Extract Agar (BAM M93) Composition per liter: Malt extract 30.0g Agar 20.0g pH 5.5 ± 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 isolation, cultivation, and identification of heat-resistant filamentous fungi (molds) from foods. Malt Extract Agar, Blakeslee’s Composition per liter: Malt extract 20.0g Glucose 20.0g Agar 20.0g Peptone 1.0g 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 a variety of yeasts, includ- ing Candida versatilis, Cryptococcus elinovii, Kluyveromyces marxi- anus, Pichia species, Reniforma strues, Rhodotorula mucilaginosa, Saccharomyces cerevisiae, Saccharomycopsis fibuligera, Sporobolo- myces roseus, Trichosporon nigrescens, Yarrowia lipolytica, Zygosac- charomyces rouxii, and numerous filamentous fungi. Malt Extract Agar, Half Strength (ATCC Medium 2418) Composition per liter: Agar 15.0g Malt extract 10.0g Peptone 2.5g pH 5.5 ± 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 until boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–118°C. Do not overheat or agar will not harden. Pour into sterile Petri dishes or distribute into sterile tubes. © 2010 by Taylor and Francis Group, LLC 1002 Malt Extract Agar for Yeasts and Molds Use: For the cultivation of yeasts and molds. Malt Extract Agar for Yeasts and Molds (MEAYM) (BAM M182) Composition per liter: Agar 20.0g Glucose 20.0g Malt extract 20.0g Peptone 1.0g pH 5.4 ± 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 isolation, cultivation, and identification of heat-resistant filamentous fungi (molds) from foods. Recommended for identifica- tion of Aspergillus spp. and Penicillium spp. Malt Extract Broth Composition per liter: Malt extract 6.0g Glucose 6.0g Maltose 1.8g Yeast extract 1.2g pH 4.7 ± 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. Do not over- heat—this results in darkening of the broth. Use: For the isolation, cultivation, and enumeration of yeast and fila- mentous fungi (mold). Malt Extract Broth Composition per liter: Malt extract 17.0g Mycological peptone 3.0g pH 5.4 ± 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 10 min at 15 psi pressure–115°C. Use: For the cultivation of molds and yeasts, especially for sterility testing. Malt Extract Broth Composition per liter: Malt extract, purified solids 15.0g pH 4.7 ± 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–118°C. Do not over- heat. Use: For the cultivation of yeasts and molds. Malt Extract Charcoal Medium (DSMZ Medium 801) Composition per liter: Malt extract 30.0g Agar 15.0g Charcoal 3.0g pH 6.5 ± 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 10 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Boletus edulis, Xerocomus badius DSM 4436, Antrodia serialis, and other filamentous fungi. Malt Extract Glucose Agar (DSMZ Medium 735) Composition per liter: Agar 15.0g Malt extract 10.0g Glucose 5.0g 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. 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 Morchella esculenta spp., Verpa digitaliformis, Disciotis venosa, Mitrophora semilibera, Gyro- mitra spp., and Mitrophora semilibera. Malt Extract HiVeg Agar Base Composition per liter: Malt extract 30.0g Agar 15.0g Plant peptone No. 4 5.0g pH 5.4 ± 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. Adjust pH to 5.4. 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 fungi, especially yeasts. Malt Extract HiVeg Agar Base, Modified Composition per liter: Agar 15.0g Maltose 12.75g Dextrin 2.75g Plant peptone 0.78g pH 6.0 ± 0.2 at 25°C © 2010 by Taylor and Francis Group, LLC Malt Yeast Extract 50% Glucose Agar (MY50G) 1003 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. Adjust pH to 6.0. 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 yeasts and molds. Malt Extract HiVeg Broth Base Composition per liter: Malt extract 17.0g Plant peptone No. 4 3.0g pH 5.4 ± 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. Adjust pH to 5.4. Gently heat and bring to boiling. Distribute into tubes or flasks. Auto- clave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of yeasts and molds. Malt Extract Peptone Agar Composition per liter: Malt extract 30.0g Agar 15.0g Soy peptone 3.0g pH 5.6 ± 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 5.6. Gently heat and bring to boiling. Distribute into tubes or flasks. Auto- clave for 10 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Basidiobolus ranarum and Sclerophoma pityophila. Malt Extract Peptone Agar Composition per liter: Malt extract 30.0g Agar 15.0g Papaic digest of soybean meal 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 Ascobolus immersus, Aspergillus amstelodami, and Aspergillus clavatus. Malt Extract Yeast Extract 40% Glucose Agar (MY40G) Composition per liter: Glucose 400.0g Agar 12.0g Malt extract powder 12.0g Yeast extract 3.0g pH 5.5 ± 0.2 at 25°C Preparation of Medium: Add components, except glucose, to 550.0mL of distilled/deionized water. Mix thoroughly. Gently heat and bring to boiling. Bring volume to 600.0mL with distilled/deionized wa- ter. While the solution is still hot, add the glucose all at once while stir- ring to avoid formation of lumps. Autoclave for 30 min at 0 psi pressure–100°C. Use: For the isolation and cultivation of osmotolerant microorganisms from foods. Malt and Peptone Medium Composition per liter: Agar 15.0g Malt extract 10.0g Peptone 5.0g NaCl 1.0g pH 6.5 ± 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. Adjust pH to 6.5. Autoclave for 15 min at 15 psi pressure– 121°C. Use: For the cultivation and maintenance of Flavobacterium species. Malt Peptone Yeast Extract Agar See: MPY Agar Malt Yeast Agar Composition per liter: Agar 20.0g Glucose 10.0g Peptone 5.0g Malt extract 3.0g Yeast extract 3.0g 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 20 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation of Arthrobacter viscosus. Malt Yeast Extract 50% Glucose Agar (MY50G) (ATCC Medium 2093) Composition per liter: Glucose 500.0g Agar 10.0g Malt extract 10.0g Yeast extract 2.5g pH 5.5 ± 0.2 at 25°C Preparation of Medium: Add agar, yeast extract, and malt extract to distilled/deionized water and bring volume to 500mL. Mix thor- oughly. Gently heat while stirring until boiling. Slowly add glucose while stirring to avoid lumps. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Do not overheat or agar will not harden. Pour into sterile Petri dishes or leave in tubes. Note: This agar hardens very slowly. Use: For the cultivation of yeasts and molds. © 2010 by Taylor and Francis Group, LLC 1004 Malt 2% Yeast Extract Agar Malt 2% Yeast Extract Agar (MYA2) Composition per liter: Agar 20.0g Malt extract 20.0g Yeast extract 1.0g 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 of Actinomucor elegans, Actinospora mega- lospora, Agaricus bisporus, Ceratocystis perfecta, Ceratocystis cana, Cer- atocystis seticollis, Chaetomium trilaterale, Chaetomium indicum, Chaetomium seminudum, Chaetomium piluliferum, Cirrenalia macro- cephala, Kluyveromyces species, Lepista inversa, Torula dematia, Tricho- derma pseudokoningii, and other fungi. Malt 4% Yeast Extract Agar (MYA4) Composition per liter: Malt extract 40.0g Agar 20.0g Yeast extract 1.0g 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 of Arthrobotrys arthrobotryoides, Ascospha- era apis, Bettsia alvei, Ceratocystiopsis minuta, Chaetomium spino- sum, Chaetomium piluliferum, Ciboriopsis simulata, Dactylella minuta, Dactylella rhombospora, Dactylella lysipaga, Eriopeziza cae- sia, Europhium clavigerum, Issatchenkia orientalis, Moniliella suave- olens, and other fungi. Malt 4% Yeast Extract Agar with Lupine Stems (MYA4 with Lupine Stems) Composition per liter: Malt extract 40.0g Agar 20.0g Yeast extract 1.0g Lupine stems variable Preparation of Medium: Add components, except lupine stems, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute 6.0mL volumes into tubes. Cut lupine stems into 8.0cm-long pieces. Add 2–3 lupine stems per tube. Autoclave for 15 min at 15 psi pressure–121°C. Allow tubes to cool in a slanted position. Use: For the cultivation of Ceratocystiopsis minuta-bicolor, Ceratocysti- opsis retusi, Ceratocystis pilifera, Ceratocystis olivaceapini, Ceratocystis multiannulata, Ceratocystis nigra, Ceratocystis olivacea, Ceratocystis tremuloaurea, Chaetomium indicum, and Chaetospermum chaetosporum. Malt 8% Yeast Extract Agar (MYA8) Composition per liter: Malt extract 80.0g Agar 20.0g Yeast extract 1.0g 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 of Thielavia hyalocarpa. Maltea Composition per liter: Maltea 40.0g Agar 22.0g Yeast extract 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 of Leucogyrophana mollusca. Maltose Peptone Yeast Extract Agar See: MPY Agar Maltose Peptone Yeast Extract Broth See: MPY Broth Maltose Peptone Yeast Extract Medium See: MP Y Agar Manganese Acetate Agar Composition per liter: Agar, highly purified 10.0g Manganous acetate 0.1g pH 7.0 ± 0.2 at 25°C Preparation of Medium: Add manganous acetate to distilled/de- ionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.0. Add agar. Steam the medium to dissolve agar. Distribute into screw-capped tubes or bottles. Autoclave for 15 min at 15 psi pressure– 121°C. Allow tubes or bottles to cool in a slanted position. Use: For the cultivation of manganese-oxidizing bacteria. Manganese Agar No. 1 (Mn Agar No. 1) Composition per liter: Agar 10.0g MnCO 3 2.0g Beef extract 1.0g Fe(NH 4 ) 2 (SO 4 ) 2 0.15g Sodium citrate 0.15g Yeast extract 0.075g Cyanocobalamin solution 10.0mL Cyanocobalamin Solution: Composition per 10.0mL: Cyanocobalamin 0.005mg Preparation of Cyanocobalamin Solution: Add cyanocobala- min to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except cyanocobala- min, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add 10.0mL of the sterile cyanocobalamin solu- © 2010 by Taylor and Francis Group, LLC . densities of mag- netic cell are wanted, ferric quinate also can be fed. For transfer use cell material which has been concentrated at the glass wall of the culture vessel by means of a magnetic. pressure–121°C. Avoid intro- duction of carbon and nitrogen from other sources. Use: For the cultivation and differentiation of coliforms and other enteric organisms, particularly Enterobacter and Escherichia,. cultivation of a variety of yeasts and other fungi. Malt Agar 2% with Lupine Stems (MA2 with Lupine Stems) Composition per liter: Malt extract 20.0g Agar 15.0g Lupine stems variable Preparation of Medium: