Motility Test and Maintenance Medium 1225 Use: For the detection of motility in Gram-negative nonfermentative bacteria. Motility Nitrate Medium, Buffered Composition per liter: Peptone 5.0g Galactose 5.0g Agar 3.0g Beef extract 3.0g Na 2 HPO 4 2.5g KNO 3 1.0g Glycerin 5.0mL pH 7.3 ± 0.1 at 25°C Preparation of Medium: Add components, except agar, to dis- tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gen- tly heat until dissolved. Add agar. Gently heat until boiling. Distribute into tubes in 11.0mL volumes. Autoclave for 15 min at 15 psi pressure– 121°C. If not used within 4 hr, heat tubes to 100°C for 10 min. Use: For the cultivation and observation of the motility of Clostridium perfringens. Motility Sulfide Medium Composition per liter: Gelatin 80.0g Proteose peptone 10.0g NaCl 5.0g Agar 4.0g Beef extract 3.0g Sodium citrate 2.0g L-Cystine 0.2g Ferrous ammonium citrate 0.2g pH 7.3 ± 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 in 4–5.0mL volumes. Autoclave for 15 min at 10 psi pressure–116°C. Use: For the determination of bacterial motility and the ability of bac- teria to produce H 2 S from L-cystine. For the differentiation of Gram- negative bacteria of the Enterobacteriaceae. Motility Test HiVeg Medium Composition per liter: Plant hydrolysate No. 1 10.0g Agar 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 in 4.0mL volumes. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the detection of bacterial motility. Motility Test HiVeg Medium (Edwards and Ewing HiVeg Medium) Composition per liter: Plant peptone 10.0g NaCl 5.0g Agar 4.0g Plant extract 3.0g 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 and bring to boiling. Distribute into tubes in 4.0mL volumes. Autoclave for 15 min at 15 psi pressure–121°C. Use: For testing motility of enteric bacteria. Motility Test HiVeg Medium with Triphenyltetrazolium Chloride (Edwards and Ewing HiVeg Medium) Composition per liter: Plant peptone 10.0g NaCl 5.0g Agar 4.0g Plant extract 3.0g Triphenyltetrazolium chloride solution 5.0mL pH 7.2 ± 0.2 at 25°C Source: This medium, without triphenyltetrazolium chloride solution, is available as a premixed powder from HiMedia. Triphenyltetrazolium Choride Solution: Composition per 5.0mL: Triphenyltetrazolium chloride 0.1g Preparation of Triphenyltetrazolium Choride Solution: Add triphenyltetrazolium chloride to distilled/deionized water and bring volume to 5.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except triphenyltetra- zolium chloride solution, 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. Aseptically add 5.0mL triphenyltetrazolium chloride so- lution. Mix thoroughly. Aseptically distribute into tubes. Motility Test and Maintenance Medium Composition per liter: Peptone 10.0g NaCl 5.0g Agar 4.0g Beef extract 3.0g 2,3,5-Triphenyltetrazolium chloride 0.05g Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into screw- capped tubes in 8.0mL volumes. Autoclave for 15 min at 15 psi pres- sure–121°C. Use: For the cultivation, maintenance, and observation of the motility of Listeria monocytogenes. © 2010 by Taylor and Francis Group, LLC 1226 Motility Test and Maintenance Medium Motility Test and Maintenance Medium Composition per liter: Agar 9.0g Tryptose 8.0g NaCl 5.0g Pancreatic digest of gelatin 2.5g Beef extract 1.5g pH 7.2 ± 0.1 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 in 7.0mL volumes. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Pass the cooled tubes into an anaerobic chamber containing 85% N 2 + 10% H 2 + 5% CO 2 . Use: For the cultivation, maintenance, and observation of the motility in a variety of anaerobic bacteria. Motility Test and Maintenance Medium Composition per liter: Peptone 10.0g NaCl 5.0g Agar 4.0g Beef extract 3.0g pH 7.4 ± 0.1 at 25°C Preparation of Medium: Add components to distilled/deionized wa- ter and bring volume to 1.0L. Mix thoroughly. Distribute into screw-capped tubes in 8.0mL volumes. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation, maintenance, and observation of motility in members of the Enterobacteriaceae. Motility Test and Maintenance Medium, Gilardi Composition per liter: Pancreatic digest of casein 10.0g NaCl 5.0g Agar 3.0g Yeast extract 3.0g pH 7.2 ± 0.1 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 3.5mL volumes. Autoclave for 15 min at 15 psi pres- sure–121°C. Use: For the cultivation, maintenance, and observation of motility in nonfermenting Gram–negative bacteria. Motility Test and Maintenance Medium, Tatum Composition per liter: Tryptose 8.0g NaCl 5.0g Agar 4.0g Pancreatic digest of gelatin 2.5g Beef extract 1.5g 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 screw- capped tubes in 8.0mL volumes. Autoclave for 15 min at 15 psi pres- sure–121°C. Use: For the cultivation, maintenance, and observation of motility in nonfermenting Gram–negative bacteria. Motility Test Medium Composition per liter: Pancreatic digest of gelatin 10.0g NaCl 5.0g Agar 4.0g Beef extract 3.0g Triphenyltetrazolium chloride solution 5.0mL pH 7.3 ± 0.2 at 25°C Source: This medium is available as a premixed powder from BD Di- agnostic Systems. 2,3,5-Triphenyltetrazolium Chloride Solution: Composition per 10.0mL: 2,3,5-Triphenyltetrazolium chloride 0.1g Preparation of 2,3,5-Triphenyltetrazolium Chloride Solu- tion: Add 2,3,5-triphenyltetrazolium chloride to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components to distilled/deionized water and bring volume to 995.0mL. Mix thoroughly. Gently heat while stirring and bring to boiling. Autoclave for 15 min at 15 psi pres- sure–121°C. Cool to 45°–50°C. Aseptically add 5.0mL of sterile 2,3,5- triphenyltetrazolium chloride solution. Mix thoroughly. Aseptically distribute into sterile tubes. Use: For detection of the motility of Gram-negative enteric bacteria. Motility Test Medium Composition per liter: Tryptose 10.0g NaCl 5.0g Agar 5.0g pH 7.2 ± 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 in 4–5.0mL volumes. Autoclave for 15 min at 15 psi pressure–121°C. Cool tubes quickly in an upright position. Use: For the determination of bacterial motility. Motility Test Medium, Semisolid Composition per liter: Peptone 10.0g NaCl 5.0g Agar 4.0g Beef extract 3.0g 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 while stirring and bring to boiling. Distribute into screw-capped tubes in 8.0mL or 20.0mL volumes. Autoclave for 15 min at 15 psi pressure– 121°C. Pour into sterile Petri dishes in 20.0mL volumes or leave in tubes. © 2010 by Taylor and Francis Group, LLC MP 5 Medium 1227 Use: For the cultivation and observation of motility in a variety of bac- teria, especially Salmonella species. Motility Test Medium, Semisolid with Sodium Chloride (BAM M103) Composition per liter: NaCl 25.0g Peptone 10.0g Agar 4.0g Beef extract 3.0g 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 while stirring and bring to boiling. Distribute into screw-capped tubes in 8.0mL volumes. Autoclave for 15 min at 15 psi pressure–121°C. Leave in tubes. Use: For the cultivation and observation of motility in a variety of bac- teria, especially Salmonella spp. and Listeria spp. For Listeria spp. keep tubes tightly screw capped and sealed with parafilm. For Salmonella spp. use Petri plates prepared on the same day as use. Use: For the cultivation of yeasts and fungi. MOX Agar See: Magnesium Oxalate Agar See: Oxford Agar, Modified MOX HiVeg Agar Composition per liter: Agar 15.0g Plant hydrolysate 15.0g Papaic digest of soybean meal 5.0g NaCl 5.0g MnCl 2 4.067g Sodium oxalate 2.68g pH 7.5 ± 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 Yersinia enterocolitica from foods. MP Agar Composition per liter: Agar 15.0g Sodium acetate 0.1g Basal medium 1.0L Sodium sulfide solution 3.0mL pH 7.0–7.5 at 25°C Basal Medium: Composition per liter: CaSO 4 ·2H 2 O (saturated solution) 20.0mL NH 4 Cl (4% solution) 5.0mL Trace elements solution 5.0mL MgSO 4 ·7H 2 O (1% solution) 1.0mL K 2 HPO 4 (1% solution) 1.0mL Preparation of Basal Medium: Add components to distilled/de- ionized water and bring volume to 1.0L. Mix thoroughly. Trace Elements Solution: Composition per liter: Ferrous EDTA solution 20.0mL ZnSO 4 ·7H 2 O (0.1% solution) 10.0mL MnSO 4 ·4H 2 O (0.02% solution) 10.0mL CuSO 4 ·5H 2 O (0.00005% solution) 10.0mL H 3 BO 3 (0.1% solution) 10.0mL Co(NO 3 ) 2 or CoCl 2 ·6H 2 O (0.01% solution) 10.0mL Na 2 MoO 4 ·2H 2 O (0.01% solution) 10.0mL Preparation of Trace Elements Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Ferrous EDTA Solution: Composition per 100.0mL: FeSO 4 ·7H 2 O 7.0g EDTA 2.0g HCl, concentrated 1.0mL Preparation of Ferrous EDTA Solution: Add components to distilled/deionized water and bring volume to 100.0mL. Mix thorough- ly. Sodium Sulfide Solution: Composition per 10.0mL: Na 2 S·9H 2 0 1.0g Preparation of Sodium Sulfide Solution: Add Na 2 S·9H 2 0 to dis- tilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Prepare freshly. Preparation of Medium: Add sodium acetate and agar to 1.0L of basal medium. Mix thoroughly. Adjust pH to 7.0–7.5. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add 3.0mL of sterile sodium sulfide solution immediately prior to dispensing. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile screw-capped tubes. Use: For the isolation and cultivation of Beggiatoa species and myx- otrophic strains of Thiothrix species from water and environmental sources. MP 5 Medium (Mineral Pectin 5 Medium) Composition per liter: Agar solution 500.0mL Basal medium 250.0mL Mineral solution 250.0mL pH 5.0–6.0 at 25°C Agar Solution: Composition per 500.0mL: Agar 15.0g Preparation of Agar Solution: Add agar to distilled/deionized water and bring volume to 500.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Basal Medium: Composition per 250.0mL: Na 2 HPO 4 6.0g Pectin, citrus or apple 5.0g © 2010 by Taylor and Francis Group, LLC 1228 MP 7 Medium KH 2 PO 4 4.0g NH 4 SO 4 2.0g Yeast extract 1.0g Preparation of Basal Medium: Add components to distilled/de- ionized water and bring volume to 250.0mL. Mix thoroughly. Gently heat and bring to boiling. Mineral Solution: Composition per 250.0mL: FeSO 4 (0.1% solution) 1.0mL MgSO 4 ·7H 2 O (20% solution) 1.0mL CaCl 2 ·2H 2 O (0.1% solution) 1.0mL H 3 BO 3 (0.001% solution) 1.0mL MnSO 4 ·H 2 O (0.001% solution) 1.0mL ZnSO 4 ·7H 2 O (0.007% solution 1.0mL CuSO 4 ·5H 2 O (0.005% solution) 1.0mL MoO 3 (0.001% solution) 1.0mL Preparation of Mineral Solution: Add components to distilled/ deionized water and bring volume to 250.0mL. Mix thoroughly. Preparation of Medium: Combine 250.0mL of basal medium and 250.0mL of mineral solution. Mix thoroughly. Adjust pH to 5.0–6.0 with 1N HCl. Autoclave the basal medium-mineral solution and agar solution separately for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Asep- tically combine the two sterile solutions. Mix thoroughly. Pour immedi- ately into sterile Petri dishes to prevent hydrolysis of the agar. Use: For the cultivation of microorganisms that produce polygalacta- nase. MP 7 Medium (Mineral Pectin 7 Medium) Composition per liter: Basal medium 500.0mL Mineral solution 500.0mL pH 7.2 ± 0.2 at 25°C Basal Medium: Composition per 500.0mL: Agar 15.0g Na 2 HPO 4 6.0g Pectin (citrus or apple) 5.0g KH 2 PO 4 4.0g NH 4 SO 4 2.0g Yeast extract 1.0g Preparation of Basal Medium: Add components to distilled/de- ionized water and bring volume to 500.0mL. Mix thoroughly. Gently heat and bring to boiling. Mineral Solution: Composition per 500.0mL: FeSO 4 (0.1% solution) 1.0mL MgSO 4 ·7H 2 O (20% solution) 1.0mL CaCl 2 ·2H 2 O (0.1% solution) 1.0mL H 3 BO 3 (0.001% solution) 1.0mL MnSO 4 ·H 2 O (0.001% solution) 1.0mL ZnSO 4 ·7H 2 O (0.007% solution 1.0mL CuSO 4 ·5H 2 O (0.005% solution 1.0mL MoO 3 (0.001% solution) 1.0mL Preparation of Mineral Solution: Add components to distilled/ deionized water and bring volume to 500.0mL. Mix thoroughly. Preparation of Medium: Combine 500.0mL of basal medium and 500.0mL of mineral solution. Mix thoroughly. Adjust pH to 7.2. Auto- clave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Pour into sterile Petri dishes. Use: For the cultivation of microorganisms that produce pectate lyase. m-PA Agar See: PA Agar m-PA-C Agar See: PA-C Agar MPH Agar (Milk Protein Hydrolysate Agar) Composition per liter: Agar 15.0g Casein hydrolysate 9.0g Glucose 1.0g pH 7.0 ± 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 while stirring and bring to boiling. Autoclave for 15 min at 15 psi pressure– 121°C. Aseptically distribute into sterile tubes. Cool to 43°–45°C be- fore using. Use: For use in the enumeration of bacteria in water and dairy prod- ucts. MPHM Medium Composition per 1275.0mL: McCoy's 5A medium, modified 1.0L Peptone solution 250.0mL Hemin solution 25.0mL Source: McCoy’s 5A medium, modified, is available from Gibco. McCoys 5A Medium, Modified: Composition per liter: Inorganic salt solution 400.0mL Other component solution 400.0mL Amino acid solution 100.0mL Vitamin solution 100.0mL Inorganic Salt Solution: Composition per 400.0mL: NaCl 6.46g NaHCO 3 2.2g NaH 2 PO 4 ·H 2 O 0.58g KCl 0.4g CaCl 2 , anhydrous 0.1g MgSO 4 , anhydrous 97.67mg Preparation of Inorganic Salt Solution: Add components to dis- tilled/deionized water and bring volume to 400.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Other Component Solution: Composition per 400.0mL: D-Glucose 3.0g Peptone 0.6g © 2010 by Taylor and Francis Group, LLC MPOB Medium 1229 Phenol Red 0.1g Glutathione, reduced 0.5mg Preparation of Other Component Solution: Add components to distilled/deionized water and bring volume to 400.0mL. Mix thor- oughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Amino Acid Solution: Composition per 100.0mL: L-Glutamine 219.2mg L-Asparagine 45.0mg L-Arginine·HCl 42.1mg L-Isoleucine 39.3mg L-Leucine 39.3mg L-Lysine·HCl 36.5mg L-Cysteine 31.5mg L-Serine 26.3mg L-Tyrosine·2Na·2H 2 O 26.2mg L-Glutamic acid 22.1mg L-Histidine·HCl·H 2 O 21.0mg L-Aspartic acid 20.0mg L-Hydroxyproline 19.7mg L-Threonine 17.9mg L-Valine 17.6mg L-Proline 17.3mg L-Phenylalanine 16.5mg L-Methionine 15.0mg L-Alanine 13.9mg Glycine 7.5mg L-Tryptophan 3.1mg Preparation of Amino Acid Solution: Add components to dis- tilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Vitamin Solution: Composition per 100.0mL: i-Inositol 36.0mg Folic acid 10.0mg Choline chloride 5.0mg Vitamin B 12 2.0mg p-Aminobenzoic acid 1.0mg Ascorbic acid 0.5mg Niacinamide 0.5mg Nicotinic acid 0.5mg Pyridoxal·HCl 0.5mg Pyridoxine·HCl 0.5mg Biotin 0.2mg D-Ca pantothenate 0.2mg Riboflavin 0.2mg Thiamine·HCl 0.2mg Preparation of Vitamin Solution: Add components to distilled/ deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Preparation of McCoys 5A Medium, Modified: Aseptically combine 400.0mL of sterile inorganic salt solution, 400.0mL of sterile other component solution, 100.0mL of sterile amino acid solution, and 100.0mL of sterile vitamin solution. Peptone Solution: Composition per 300.0mL: Proteose peptone No. 3 7.5g Preparation of Peptone Solution: Add components to distilled/ deionized water and bring volume to 300.0mL. Mix thoroughly. Auto- clave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Hemin Solution: Composition per 100.0mL: NaCl 0.8g Na 2 HPO 4 0.12g Hemin 0.05g KCl 0.04g MgCl 2 ·6H 2 O 0.03g NaH 2 PO 4 ·H 2 O 0.02g CaCl 2 0.011g Preparation of Hemin Solution: Add NaCl, KCl, NaH 2 PO 4 ·H 2 O, and Na 2 HPO 4 to distilled/deionized water and bring volume to 50.0mL. Mix thoroughly. Separately add the CaCl 2 and MgCl 2 ·6H 2 O to distilled/deionized water and bring volume to 50.0mL. Mix thor- oughly. Combine the two solutions. Mix thoroughly. Add 0.05g of hemin. Mix thoroughly. Adjust to pH 9.0 with 10N NaOH to dissolve the hemin. Mix thoroughly. Filter sterilize. Preparation of Medium: Aseptically combine 1.0L of sterile mod- ified McCoy’s medium 5A, 250.0mL of sterile peptone solution, and 25.0mL of sterile hemin solution. Aseptically distribute into sterile screw-capped tubes or flasks. Use: For the cultivation of Trypanosoma theileri. m-Plate Count Broth See: Plate Count Broth MPOB Medium Composition per 1012.0mL: Disodium fumarate 3.2g Na 2 HPO 4 ·2H 2 O 0.53g KH 2 PO 4 0.41g NaCl 0.3g NH 4 Cl 0.3g Yeast extract 0.2g CaCl 2 ·2H 2 O 0.11g MgCl 2 ·6H 2 O 0.10g Resazurin 0.5mg NaHCO 3 solution 20.0mL Na 2 S·9H 2 O solution 10.0mL Wolfe’s vitamin solution 10.0mL Selenite-tungstate solution 1.0mL Trace elements solution SL-10 1.0mL pH 7.0–7.2 at 25°C NaHCO 3 Solution: Composition per 20.0mL: NaHCO 3 4.0g Preparation of NaHCO 3 Solution: Add NaHCO 3 to distilled/deion- ized water and bring volume to 20.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Na 2 S·9H 2 O Solution: Composition per 10.0mL: Na 2 S·9H 2 O 0.5g Preparation of Na 2 S·9H 2 O Solution: Add Na 2 S·9H 2 O to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. © 2010 by Taylor and Francis Group, LLC 1230 MPSS Broth Wolfe’s Vitamin Solution: Composition per liter: Pyridoxine·HCl 10.0mg p-Aminobenzoic acid 5.0mg Lipoic acid 5.0mg Nicotinic acid 5.0mg Riboflavin 5.0mg Thiamine·HCl 5.0mg Calcium DL-pantothenate 5.0mg Biotin 2.0mg Folic acid 2.0mg Vitamin B 12 0.1mg Preparation of Wolfe’s Vitamin Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize. Selenite-tungstate Solution: Composition per liter: NaOH 0.5g Na 2 WO 4 ·2H 2 O 4.0mg Na 2 SeO 3 ·5H 2 O 3.0mg Preparation of Selenite-tungstate Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Trace Elements Solution SL-10: Composition per liter: FeCl 2 ·4H 2 O 1.5g CoCl 2 ·6H 2 O 190.0mg MnCl 2 ·4H 2 O 100.0mg ZnCl 2 70.0mg Na 2 MoO 4 ·2H 2 O 36.0mg NiCl 2 ·6H 2 O 24.0mg H 3 BO 3 6.0mg CuCl 2 ·2H 2 O 2.0mg HCl (25% solution) 10.0mL Preparation of Trace Elements Solution SL-10: Add FeCl 2 ·4H 2 O to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized water and bring volume to 1.0L. Add remaining components. Mix thor- oughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure– 121°C. Preparation of Medium: Prepare medium under 80% N 2 + 20% CO 2 . Add components, except NaHCO 3 solution and Na 2 S·9H 2 O solu- tion, to distilled/deionized water and bring volume to 970.0mL. Mix thor- oughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add 20.0mL of sterile NaHCO 3 solution and 10.0mL of sterile Na 2 S·9H 2 O solution. Mix thor- oughly. Aseptically and anaerobically distribute into sterile tubes or flasks. After inoculation, bring culture bottles to 0.7 bar 80% N 2 + 20% CO 2 over- pressure. Use: For the cultivation of Syntrophobacter species. m-Pseudomonas aeruginosa Agar See: Pseudomonas aeruginosa Agar MPSS Broth Composition per liter: Peptone 5.0g MgSO 4 ·7H 2 O 1.0g (NH 4 ) 2 SO 4 1.0g Succinic acid 1.0g FeCl 3 ·6H 2 O (0.2% solution) 1.0mL MnSO 4 ·H 2 O (0.2% solution) 1.0mL 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 Spirillum volutans. MPY Agar (Malt Peptone Yeast Extract Agar) (ATCC Medium 582) Composition per liter: Agar 15.0g Malt extract 5.0g Xylose 2.0g Fructose 2.0g Lactose 2.0g Peptone 1.0g Yeast extract 1.0g 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 Streptomyces naniwaen- sis. MPY Agar (Maltose Peptone Yeast Extract Medium) (ATCC Medium 518) Composition per liter: Agar 10.0g Maltose 2.0g Peptone 2.0g Yeast extract 1.0g Potassium phosphate buffer (1M, pH 7.5) 10.0mL pH 7.5 ± 0.2 at 25°C Preparation of Medium: Add components, except potassium phosphate buffer, 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. Filter sterilize potassium phosphate buffer. Aseptically add sterile potassium phos- phate buffer to sterile, cooled basal medium. Distribute into sterile tubes or flasks. Use: For the cultivation and maintenance of Spirochaeta aurantia. MPY Broth (Maltose Peptone Yeast Extract Broth) Composition per liter: Maltose 2.0g Peptone 2.0g Yeast extract 1.0g Potassium phosphate buffer (1M, pH 7.5) 10.0mL pH 7.5 ± 0.2 at 25°C Preparation of Medium: Add components, except potassium phosphate buffer, to distilled/deionized water and bring volume to © 2010 by Taylor and Francis Group, LLC MRS Agar with 0.5% Cysteine 1231 990.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure– 121°C. Filter sterilize potassium phosphate buffer. Aseptically add sterile potassium phosphate buffer to sterile cooled basal medium. Dis- tribute into sterile tubes or flasks. Use: For the cultivation and maintenance of Spirochaeta aurantia. MPYG Medium See: Peptone Yeast Extract Glucose Medium, Modified MRS Agar (DeMan, Rogosa, Sharpe Agar) Composition per liter: Glucose 20.0g Peptone 10.0g Agar 10.0g Beef extract 8.0g Sodium acetate·3H 2 O 5.0g Yeast extract 4.0g K 2 HPO 4 2.0g Triammonium citrate 2.0g MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 0.05g Sorbitan monooleate 1.0mL pH 6.2 ± 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. 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 lactic acid bacteria. MRS Agar (DeMan, Rogosa, Sharpe Agar) Composition per liter: Glucose 18.5g Agar 13.5g Pancreatic digest of gelatin 10.0g Beef extract 8.0g Yeast extract 4.0g Sodium acetate 3.0g K 2 HPO 4 2.0g Ammonium citrate 2.0g Polysorbate 80 1.0g MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 0.05g pH 6.2 ± 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 isolation and cultivation of Lactobacillus species from clinical specimens, foods, and dairy products. MRS Agar with Cysteine (LMG Medium 118) Composition per liter: Glucose 20.0g Peptone 10.0g Agar 12.0g Beef extract 5.0g Sodium acetate·3H 2 O 5.0g Yeast extract 5.0g KH 2 PO 4 2.0g Diammonium hydrogen citrate 2.0g MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 38.0mg L-Cysteine solution 10.0mL Tween™ 80 1.0mL pH 6.5 ± 0.2 at 25°C L-Cysteine Solution: Composition per 10.0mL: L-Cysteine 0.5g Preparation of L-Cysteine Solution: Add 0.1g of L-cysteine to distilled/deionized water and bring volume to 10.0mL. Mix thorough- ly. Filter sterilize. Preparation of Medium: Add components, except L-cysteine solu- tion, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Adjust pH to 6.5. Gently heat and bring to boiling. Distrib- ute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Aseptically add 10.0mL of sterile L-cysteine solution. Mix thoroughly. Aseptically pour into sterile Petri dishes or distribute into sterile tubes. Use: For the isolation and cultivation of Leuconostoc mesenteroides and other lactic acid bacteria. MRS Agar with 0.5% Cysteine (LMG Medium 131) Composition per liter: Glucose 20.0g Agar 12.0g Peptone 10.0g Beef extract 5.0g Sodium acetate·3H 2 O 5.0g Yeast extract 5.0g KH 2 PO 4 2.0g Diammonium hydrogen citrate 2.0g MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 38.0mg L-Cysteine·HCl solution 100.0mL Tween™ 80 1.0mL pH 6.5 ± 0.2 at 25°C L-Cysteine·HCl Solution: Composition per 100.0mL: L-Cysteine·HCl 5.0g Preparation of L-Cysteine·HCl Solution: Add 5.0g of L-cyste- ine·HCl to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except L-cysteine·HCl solution, to distilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Adjust pH to 6.5. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure– © 2010 by Taylor and Francis Group, LLC 1232 MRS Agar with Ethanol 121°C. Cool to 50°C. Aseptically add 100.0mL of sterile L- cysteine·HCl solution. Mix thoroughly. Aseptically pour into sterile Petri dishes or distribute into sterile tubes. Use: For the isolation and cultivation of Lactobacillus ruminis and Lactobacillus vitulinus. MRS Agar with Ethanol (LMG Medium 130) Composition per liter: Glucose 20.0g Agar 12.0g Peptone 10.0g Beef extract 5.0g Sodium acetate·3H 2 O 5.0g Yeast extract 5.0g KH 2 PO 4 2.0g Diammonium hydrogen citrate 2.0g MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 38.0mg Ethanol 100.0mL Tween™ 80 1.0mL pH 5.0± 0.2 at 25°C Preparation of Medium: Add components, except ethanol, to dis- tilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Asepti- cally add 100.0mL of sterile ethanol. Mix thoroughly. Aseptically dis- tribute into sterile tubes or pour into sterile Petri dishes. Use: For the cultivation of Lactobacillus acetotolerans. MRS Agar with Tomato Juice, pH 5.2 (LMG Medium 248) Composition per liter: Glucose 20.0g Peptone 10.0g Agar 12.0g Beef extract 5.0g Sodium acetate·3H 2 O 5.0g Yeast extract 5.0g KH 2 PO 4 2.0g Diammonium hydrogen citrate 2.0g MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 38.0mg Tomato juice 100.0mL Tween™ 80 1.0mL pH 5.2 ± 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 5.2 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 isolation and cultivation of Lactobacillus kunkeei. MRS Agar, Half Strength (LMG Medium 281) Composition per liter: Glucose 10.0g Agar 6.0g Peptone 5.0g Beef extract 2.5g Sodium acetate·3H 2 O 2.5g Yeast extract 2.5g KH 2 PO 4 1.0g Diammonium hydrogen citrate 1.0g MgSO 4 ·7H 2 O 0.1g MnSO 4 ·4H 2 O 19.0mg Tween™ 80 0.5mL 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. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of Lactobacillus acidipiscis and Weissella thai- landensis. MRS Broth Composition per liter: Glucose 20.0g Peptone 10.0g Beef extract 5.0g Sodium acetate·3H 2 O 5.0g Yeast extract 5.0g K 2 HPO 4 2.0g Diammonium hydrogen citrate 2.0g MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 38.0mg Sorbitan monooleate 1.0mL pH 6.2 ± 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. Use: For the cultivation of Leuconostoc pseudomesenteroides, Leu- conostoc carnosum, and Lactobacillus crispatus. MRS Broth (DeMan, Rogosa, Sharpe Broth) Composition per liter: Glucose 20.0g Peptone 10.0g Beef extract 8.0g Sodium acetate·3H 2 O 5.0g Yeast extract 4.0g K 2 HPO 4 2.0g Triammonium citrate 2.0g MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 0.05g Sorbitan monooleate 1.0mL pH 6.2 ± 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. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of lactic acid bacteria. © 2010 by Taylor and Francis Group, LLC MRS Broth with Ethanol 1233 MRS Broth (DeMan, Rogosa, Sharpe Broth) Composition per liter: Glucose 18.5g Pancreatic digest of gelatin 10.0g Beef extract 8.0g Yeast extract 4.0g Sodium acetate 3.0g K 2 HPO 4 2.0g Ammonium citrate 2.0g Polysorbate 80 1.0g MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 0.05g pH 6.2 ± 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 isolation and cultivation of Lactobacillus species from clinical specimens, foods, and dairy products. MRS Broth with CaCO 3 (LMG Medium 166) Composition per liter: CaCO 3 30.0g Glucose 20.0g Peptone 10.0g Beef extract 5.0g Sodium acetate·3H 2 O 5.0g Yeast extract 5.0g K 2 HPO 4 2.0g Diammonium hydrogen citrate 2.0g MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 38.0mg Sorbitan monooleate 1.0mL pH 6.2 ± 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. Use: For the cultivation of Sporolactobacillus inulinus. MRS Broth with Cysteine Composition per liter: Glucose 18.5g Pancreatic digest of gelatin 10.0g Beef extract 8.0g Yeast extract 4.0g Sodium acetate 3.0g K 2 HPO 4 2.0g Ammonium citrate 2.0g Polysorbate 80 1.0g MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 0.05g L-Cysteine solution 0.5mL pH 6.2 ± 0.2 at 25°C L-Cysteine Solution: Composition per 10.0mL: L-Cysteine 0.1g Preparation of L-Cysteine Solution: Add 0.1g of L-cysteine to distilled/deionized water and bring volume to 10.0mL. Mix thorough- ly. Filter sterilize. Preparation of Medium: Add components, except L-cysteine solu- tion, to distilled/deionized water and bring volume to 999.5mL. Mix thoroughly. Adjust pH to 6.2. Autoclave for 15 min at 15 psi pressure– 121°C. Aseptically add 0.5mL of sterile L-cysteine solution. Mix thor- oughly. Aseptically distribute into sterile tubes or flasks. Use: For the isolation and cultivation of Leuconostoc mesenteroides. MRS Broth with Ethanol (LMG Medium 240) Composition per liter: Glucose 20.0g Peptone 10.0g Beef extract 5.0g Sodium acetate·3H 2 O 5.0g Yeast extract 5.0g KH 2 PO 4 2.0g Diammonium hydrogen citrate 2.0g MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 38.0mg Ethanol 100.0mL Tween™ 80 1.0mL pH 5.0± 0.2 at 25°C Preparation of Medium: Add components, except ethanol, to dis- tilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Asepti- cally add 100.0mL of sterile ethanol. Mix thoroughly. Aseptically dis- tribute into sterile tubes or sterile flasks. Use: For the cultivation of Lactobacillus acetotolerans. MRS Broth with Ethanol Composition per liter: Glucose 20.0g Peptone 10.0g Beef extract 8.0g Sodium acetate·3H 2 O 5.0g Yeast extract 4.0g K 2 HPO 4 2.0g Triammonium citrate 2.0g MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 0.05g Sorbitan monooleate 1.0mL Ethanol (95% solution), filter sterilized 100.0mL pH 5.0 ± 0.2 at 25°C Preparation of Medium: Add components, except ethanol, to dis- tilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically add 100.0mL of sterile ethanol. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation of Lactobacillus acetotolerans. © 2010 by Taylor and Francis Group, LLC 1234 MRS Chalk MRS Chalk Composition per liter: CaCO 3 30.0g Glucose 20.0g Beef extract 10.0g Peptone 10.0g Yeast extract 5.0g K 2 HPO 4 2.0g Triammonium citrate 2.0g MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 0.05g Tween™ 80™ 1.0mL pH 6.2 ± 0.4 at 25°C Preparation of Medium: Add components to distilled/deionized wa- ter and bring volume to 1.0L. Mix thoroughly. Adjust pH to 6.2. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of Sporolactobacillus inulinus and other Spor- olactobacillus species. MRS with Fermented Wort Composition per liter: Solution A 800.0mL Solution B 200.0mL pH 6.2 ± 0.2 at 25°C Solution A: Composition per liter: Glucose 20.0g Peptone 10.0g Agar 10.0g Beef extract 8.0g Sodium acetate·3H 2 O 5.0g Yeast extract 4.0g K 2 HPO 4 2.0g Triammonium citrate 2.0g MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 0.05g Sorbitan monooleate 1.0mL 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. Cool to 50°–55°C. Solution B: Composition per liter: Malt extract 15.0g Maltose 12.75g Dextrin 2.75g Glycerol 2.35g K 2 HPO 4 1.0g NH 4 Cl 1.0g Pancreatic digest of gelatin 0.78g Preparation of Solution B: 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. Cool to room temperature. Inoculate with Saccharomyces cerevisiae to allow wort to ferment. Incubate for 24–48 hr. Allow to cool to room temper- ature. Filter through Whatman filter paper to remove solids. Filter ster- ilize. Preparation of Medium: Aseptically combine 800.0mL of sterile solution A and 200.0mL of sterile solution B. Mix thoroughly. Asepti- cally distribute into sterile tubes or flasks. Use: For the cultivation of yeasts and fungi. MRS Fructose Medium Composition per liter: Beef extract 10.0g Fructose 10.0g Peptone 10.0g Yeast extract 5.0g K 2 HPO 4 2.0g Triammonium citrate 2.0g MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 0.05g Tween™ 80 1.0mL pH 6.2 ± 0.4 at 25°C Preparation of Medium: Add components to distilled/deionized wa- ter and bring volume to 1.0L. Mix thoroughly. Adjust pH to 6.2. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of Carnobacterium divergens. MRS HiVeg Agar (Lactobacillus MRS HiVeg Agar) Composition per liter: Glucose 20.0g Peptone 10.0g Agar 10.0g Plant extract 8.0g Sodium acetate·3H 2 O 5.0g Yeast extract 4.0g K 2 HPO 4 2.0g Triammonium citrate 2.0g MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 0.05g pH 6.2 ± 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 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 lactic acid bacteria. MRS HiVeg Agar, Modified (Lactobacilli Heteroferm Screen HiVeg Agar) Composition per liter: Glucose 20.0g Agar 15.0g Plant peptone No. 3 10.0g Sodium acetate 5.0g Yeast extract 5.0g 2-Phenylethyl alcohol 3.0g Ammonium citrate 2.0g K 2 HPO 4 2.0g MgSO 4 0.1g MnSO 4 0.05g © 2010 by Taylor and Francis Group, LLC . Use: For the determination of bacterial motility and the ability of bac- teria to produce H 2 S from L-cystine. For the differentiation of Gram- negative bacteria of the Enterobacteriaceae. Motility. 1.0mL Preparation of Mineral Solution: Add components to distilled/ deionized water and bring volume to 250.0mL. Mix thoroughly. Preparation of Medium: Combine 250.0mL of basal medium and 250.0mL of mineral. 1.0mL Preparation of Mineral Solution: Add components to distilled/ deionized water and bring volume to 500.0mL. Mix thoroughly. Preparation of Medium: Combine 500.0mL of basal medium and 500.0mL of mineral