Lactobacillus rimae Medium 925 Na-acetate·3H 2 O 5.0g Glucose 4.0g Maltose 4.0g Meat extract 3.0g K 2 HPO 4 ·3H 2 O 2.6g (NH 4 ) 2 citrate 2.0g Cysteine-HCl·H 2 O 0.5g MgSO 4 ·7H 2 O 0.1g MnSO 4 ·4H 2 O 0.05g Tween™ 80 1.0mL pH 6.3 ± 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 Lactobacillus panis. Lactobacillus 8664 Medium Composition per liter: Maltose 20.0g Peptone 10.0g Yeast extract 10.0g Glucose 5.0g 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 brevis. Lactobacillus MRS HiVeg Agar (MRS HiVeg Agar) Composition per liter: Glucose 20.0g Agar 12.0g Plant extract 10.0g Plant peptone No. 3 10.0g Sodium acetate 5.0g Yeast extract 5.0g Ammonium citrate 2.0g K 2 HPO 4 2.0g Polysorbate 80 1.0g MgSO 4 0.1g MnSO 4 0.05g pH 6.5 ± 0.2 at 25°C Source: This medium is available as a premixed powder from HiMe- dia. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0mL. Mix thoroughly. Gently heat and bring to boiling. Adjust pH to 6.5. Autoclave for 15 min at 15 psi pres- sure–121°C. Pour into sterile Petri dishes. Use: For the cultivation of Lactobacillus species. Lactobacillus MRS HiVeg Broth (MRS HiVeg Broth) Composition per liter: Glucose 20.0g Plant extract 10.0g Plant peptone No. 3 10.0g Sodium acetate 5.0g Yeast extract 5.0g Ammonium citrate 2.0g K 2 HPO 4 2.0g Polysorbate 80 1.0g MgSO 4 0.1g MnSO 4 0.05g pH 6.5 ± 0.2 at 25°C Source: This medium is available as a premixed powder from HiMe- dia. 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 Lactobacillus species. Lactobacillus Orotic Acid Medium Composition per liter: Milk, peptonized 15.0g Glucose 10.0g Yeast extract 5.0g KH 2 PO 4 2.0g Sorbitan monooleate complex 1.0g Orotic acid 25.0mg D-Pantothenic acid 0.2mg Tomato juice 100.0mL L-Cysteine·HCl·H 2 O solution 7.5mL pH 6.8 ± 0.2 at 25°C L-Cysteine·HCl·H 2 O Solution: Composition per 10.0mL: L-Cysteine·HCl·H 2 O 0.15g Preparation of L-Cysteine·HCl·H 2 O Solution: Add L-cyste- ine·HCl·H 2 O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Continue boiling for 2–3 min. Distribute into bottles in 20.0mL volumes. Add 0.15mL of L-cysteine·HCl·H 2 O solution to each bottle containing 20.0mL of medium. Autoclave for 15 min at 15 psi pressure–121°C. Screw caps tightly to maintain reduced conditions. Use: For the cultivation of Lactobacillus helveticus. Lactobacillus rimae Medium Composition per liter: Yeast extract 10.0g Peptone 5.0g Pancreatic digest of casein 5.0g Glucose 5.0g (NH 4 ) 2 SO 4 0.5g L-Cysteine·HCl 0.5g Resazurin 1.0mg Mineral solution 40.0mL Fatty acid mixture 3.1mL Hemin solution 0.5mL Vitamin K 1 0.2mL pH 6.9 ± 0.2 at 25°C Mineral Solution: Composition per liter: NaHCO 3 10.0g NaCl 2.0g © 2010 by Taylor and Francis Group, LLC 926 Lactobacillus rimae Medium with Tween™ KH 2 PO 4 1.0g K 2 HPO 4 1.0g MgSO 4 ·7H 2 O 0.48g CaCl 2 ·2H 2 O 0.3g Preparation of Mineral Solution: Add components to distilled/ deionized water and bring volume to 1.0L. Mix thoroughly. Fatty Acid Mixture: Composition per 31.0mL: Acetic acid 17.0mL Propionic acid 6.0mL n-Butyric acid 4.0mL n-Valeric acid 1.0mL iso-Valeric acid 1.0mL iso-Butyric acid 1.0mL DL-2-Methylbutyric acid 1.0mL Preparation of Fatty Acid Mixture: Combine components. Mix thoroughly. Adjust pH to 7.5 with concentrated NaOH. Hemin Solution: Composition per 1.0mL: Hemin 5.0mg NaOH (1N solution) 1.0mL Preparation of Hemin Solution: Add hemin to 1.0mL of NaOH solution. Mix thoroughly. Preparation of Medium: Add components, except L-cysteine·HCl, hemin solution, and fatty acid mixture, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Continue boiling for 5 min. Cool to room temperature while sparging with 100% CO 2 . Add L-cysteine·HCl, hemin solution, and fatty acid mixture. Adjust pH to 6.9 with 8N NaOH while continuing to sparge with 100% CO 2 . After pH has been reached, sparge with 100% N 2 . Anaerobically dis- tribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation and maintenance of Acetivibrio ethanolgig- nens, Lactobacillus rimae, Lactobacillus uli, and Sphaerotilus natans. Lactobacillus rimae Medium with Tween™ Composition per liter: Yeast extract 10.0g Peptone 5.0g Pancreatic digest of casein 5.0g Glucose 5.0g (NH 4 ) 2 SO 4 0.5g L-Cysteine·HCl 0.5g Tween™ 80 0.2g Resazurin 1.0mg Mineral solution 40.0mL Fatty acid mixture 3.1mL Hemin solution 0.5mL Vitamin K 1 0.2mL pH 6.9 ± 0.2 at 25°C Mineral Solution: Composition per liter: NaHCO 3 10.0g NaCl 2.0g KH 2 PO 4 1.0g K 2 HPO 4 1.0g MgSO 4 ·7H 2 O 0.48g CaCl 2 ·2H 2 O 0.3g Preparation of Mineral Solution: Add components to distilled/ deionized water and bring volume to 1.0L. Mix thoroughly. Fatty Acid Mixture: Composition per 31.0mL: Acetic acid 17.0mL Propionic acid 6.0mL n-Butyric acid 4.0mL n-Valeric acid 1.0mL iso-Valeric acid 1.0mL iso-Butyric acid 1.0mL DL-2-Methylbutyric acid 1.0mL Preparation of Fatty Acid Mixture: Combine components. Mix thoroughly. Adjust pH to 7.5 with concentrated NaOH. Hemin Solution: Composition per 1.0mL: Hemin 5.0mg NaOH (1N solution) 1.0mL Preparation of Hemin Solution: Add hemin to 1.0mL of NaOH solution. Mix thoroughly. Preparation of Medium: Add components, except L-cysteine·HCl, hemin solution, and fatty acid mixture, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boil- ing. Continue boiling for 5 min. Cool to room temperature while sparg- ing with 100% CO 2 . Add L-cysteine·HCl, hemin solution, and fatty acid mixture. Adjust pH to 6.9 with 8N NaOH while continuing to sparge with 100% CO 2 . After pH has been reached, sparge with 100% N 2 . Anaerobically distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation and maintenance of Lactobacillus rimae and Lactobacillus uli. Lactobacillus Selection Agar See: LBS™ Agar Lactobacillus Selection Agar Base Composition per liter: Sodium acetate 25.0g Glucose 20.0g Agar 15.0g Casein enzymic hydrolysate 10.0g KH 2 PO 4 6.0g Yeast extract 5.0g Ammonium citrate 2.0g Polysorbate 80 1.0g MgSO 4 ·7H 2 O 0.575g MnSO 4 ·2H 2 O 0.12g FeSO 3 0.034g Acetic acid, glacial 1.32mL pH 5.5 ± 0.2 at 25°C Source: This medium is available from HiMedia. Preparation of Medium: Add acetic acid to distilled/deionized wa- ter and bring volume to 1.0L. Add remaining components. Mix thor- oughly. Distribute into tubes or flasks. Gently heat and bring to boiling. Boil for 1–2 min. Do not autoclave unless storage is needed. If storage is necessary sterilize by autoclaving for 15 min at 15 psi pressure– 121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the isolation and enumeration of lactobacilli from foods. © 2010 by Taylor and Francis Group, LLC Lactococcus piscium Medium 927 Lactobacillus Selection Broth See: LBS™ Broth Lactobacillus Selection HiVeg Agar Base with Acetic Acid and Polysorbate Composition per liter: Sodium acetate·3H 2 O 25.0g Glucose 20.0g Agar 15.0g Plant hydrolysate 10.0g KH 2 PO 4 6.0g Yeast extract 5.0g Ammonium citrate 2.0g MgSO 4 0.575g FeSO 4 0.034g MnSO 4 0.12g Acetic acid, glacial 1.32 mL Polysorbate 80 1.0mL pH 5.5 ± 0.2 at 25°C Source: This medium, without polysorbate 80 or glacial acetic acid, 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. Adjust pH to 5.5. Do not autoclave. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the selective isolation, cultivation, and enumeration of lacto- bacilli from foods. Lactobacillus Selection HiVeg Broth Base with Acetic Acid and Polysorbate Composition per liter: Sodium acetate 25.0g Glucose 20.0g Plant hydrolysate 10.0g KH 2 PO 4 6.0g Yeast extract 5.0g Polysorbate 80 1.0g MgSO 4 0.575g MnSO 4 0.12g FeSO 4 0.034g Acetic acid, glacial 1.32 mL Polysorbate 80 1.0mL pH 5.5 ± 0.2 at 25°C Source: This medium, without polysorbate 80 or glacial acetic acid, 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. Adjust pH to 5.5. Do not autoclave. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation of lactobacilli. Lactobacillus Selection Oxgall Agar (LBS™ Oxgall Agar) Composition per liter: Sodium acetate·3H 2 O 25.0g Glucose 20.0g Agar 15.0g Pancreatic digest of casein 10.0g KH 2 PO 4 6.0g Yeast extract 5.0g Ammonium citrate 2.0g Oxgall 1.5g Polysorbate 80 1.0g MgSO 4 0.575g MnSO 4 0.12g FeSO 4 0.034g Acetic acid, glacial 1.32mL pH 5.5 ± 0.2 at 25°C Preparation of Medium: Add components, except acetic acid, to distilled/deionized water and bring volume to 998.7mL. Mix thorough- ly. Gently heat and bring to boiling. Add glacial acetic acid. Mix thor- oughly. Gently heat while stirring and bring to 90°–100°C for 2–3 min. Do not autoclave. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the selective isolation, cultivation, and enumeration of lacto- bacilli. Lactobacillus Selection Oxgall Agar Base (LBS Oxgall Agar) Composition per liter: Sodium acetate 25.0g Glucose 20.0g Agar 15.0g Casein enzymic hydrolysate 10.0g KH 2 PO 4 6.0g Yeast extract 5.0g Ammonium citrate 2.0g Oxgall 1.5g Polysorbate 80 1.0g MgSO 4 ·7H 2 O 0.575g MnSO 4 ·2H 2 O 0.12g FeSO 4 ·7H 2 O 0.034g Acetic acid, glacial 1.32mL pH 5.4 ± 0.2 at 25°C Source: This medium is available from HiMedia. Preparation of Medium: Add acetic acid to distilled/deionized wa- ter and bring volume to 1.0L. Add remaining components. Mix thor- oughly. Distribute into tubes or flasks. Gently heat and bring to boiling. Boil for 1–2 min. Do not autoclave unless storage is needed. If storage is necessary, sterilize by autoclaving for 15 min at 15 psi pressure– 121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the selective isolation, cultivation, and enumeration of lacto- bacilli. Lactobacillus-Streptococcus Differential Medium See: L-S Differential Medium Lactococcus piscium Medium Composition per liter: Glucose 10.0g Peptone 10.0g Beef extract 8.0g NaCl 5.0g Yeast extract 3.0g KH 2 PO 4 1.5g © 2010 by Taylor and Francis Group, LLC 928 Lactose Blue Agar MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 0.05g pH 6.8 ± 0.2 at 25°C Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 6.8. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure– 121°C. Use: For the cultivation of Lactococcus piscium. Lactose Blue Agar (B.T.B. Lactose Agar, Modified) Composition per liter: Lactose 15.5g Agar 13.0g NaCl 5.0g Peptic digest of animal tissue 3.5g Casein enzymic hydrolysate 3.5g Bromthymol Blue 0.04g pH 7.0 ± 0.2 at 25°C Source: This medium is available as a premixed powder from HiMe- dia. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes. Use: For the differentiation of lactose-fermenting and non-fermenting bacteria belonging to Enterobacteriaceae. Lactose Blue HiVeg Agar (B.T.B. Lactose HiVeg Agar, Modified) Composition per liter: Lactose 15.5g Agar 13.0g NaCl 5.0g Plant extract 3.5g Plant hydrolysate 3.5g Bromthymol Blue 0.04g pH 7.0 ± 0.2 at 25°C Source: This medium is available as a premixed powder from HiMe- dia. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes. Use: For the differentiation of lactose-fermenting and non-fermenting bacteria belonging to Enterobacteriaceae. Lactose Broth Composition per liter: Lactose 5.0g 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 and Oxoid Unipath. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes containing an inverted Durham tube in 10.0mL volumes. Autoclave for 12 min at 15 psi pressure–121°C. Cool broth quickly to 25°C. For test- ing water samples with 10.0mL volumes, prepare medium double strength. Use: For the detection of lactose-fermenting, Gram-negative coli- forms, as a preenrichment broth for Salmonella species, and in the study of lactose fermentation of bacteria in general. Lactose Casein Hydrolysate Medium Composition per liter: Lactose 37.5g Agar 15.0g Casein hydrolysate 3.0g KH 2 PO 4 1.0g MnSO 4 0.5g Microelements solution 2.0mL pH 6.0 ± 0.2 at 25°C Microelements Solution: Composition per liter: Fe(NO 3 ) 3 ·9H 2 O 723.5mg ZnSO 4 ·7H 2 O 439.8mg MnSO 4 ·4H 2 O 203.0mg H 2 SO 4 variable Preparation of Microelements Solution: Add components, one at a time, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Be sure one component is dissolved before adding the next. Add sulfuric acid to yield a clear solution. Preparation of Medium: Add components to distilled/deionized wa- ter and bring volume to 1.0L. Mix thoroughly. Gently heat until 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 and maintenance of Drechslera catenaria. Lactose Distillers Solubles Medium Composition per liter: Lactose 20.0g Distillers solubles 15.0g Yeast, autolyzed 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 Streptomyces avermitilis. Lactose Egg Yolk Milk Agar Composition per 1206.0mL: Lactose 12.0g Agar 1.0g Columbia blood agar base 800.0mL Skim milk 150.0mL Egg yolk emulsion, 50% 36.0mL Inhibitor solution 20.0mL Neutral Red (1% solution) 3.25mL pH 7.0 ± 0.2 at 25°C © 2010 by Taylor and Francis Group, LLC Lactose Lecithin Agar 929 Columbia Blood Agar Base: Composition per 800.0mL: Agar 15.0g Pantone 10.0g Bitone 10.0g NaCl 5.0g Tryptic digest of beef heart 3.0g Cornstarch 1.0g Preparation of Columbia Blood Agar Base: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat until boiling. Egg Yolk Emulsion, 50%: Composition per 100.0mL: Chicken egg yolks 11 Whole chicken egg 1 NaCl (0.9% solution) 50.0mL Preparation of Egg Yolk Emulsion, 50%: Soak eggs with 1:100 dilution of saturated mercuric chloride solution for 1 min. Crack eggs and separate yolks from whites. Mix egg yolks with 1 chicken egg. Beat to form emulsion. Measure 50.0mL of egg yolk emulsion and add to 50.0mL of 0.9% NaCl solution. Mix thoroughly. Filter sterilize. Warm to 45°–50°C. Inhibitor Solution: Composition per 20.0mL: Neomycin sulfate 0.18g NaN 3 0.24g Caution: Sodium azide is toxic. Azides also react with metals and disposal must be highly diluted. Preparation of Inhibitor Solution: Add neomycin sulfate and NaN 3 to distilled/deionized water and bring volume to 20.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Combine Columbia blood agar base, lac- tose, agar, and Neutral Red and bring volume to 1.0L. Adjust pH to 7.0. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 55°C. Filter ster- ilize skim milk. To 1.0L of cooled, sterile agar mixture, aseptically add 150.0mL of sterile skim milk, 36.0mL of sterile egg yolk emulsion, 50%, and 20.0mL of sterile inhibitor solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of Clostridium species. Lactose Gelatin Medium Composition per liter: Gelatin 120.0g Tryptose 15.0g Lactose 10.0g Yeast extract 10.0g Phenol Red (0.5% solution) 10.0mL pH 7.5 ± 0.2 at 25°C Preparation of Medium: Add tryptose, yeast extract, and lactose to distilled/deionized water and bring volume to 400.0mL. Mix thor- oughly. Add gelatin to distilled/deionized water and bring volume to 590.0mL. Gently heat gelatin solution while stirring and bring to 50°– 60°C. Add Phenol Red. Mix the two solutions together. Distribute into tubes in 10.0mL volumes. Autoclave for 10 min at 15 psi pressure– 121°C. If medium is not used in 8 hr, deoxygenate by heating to 50°– 70°C for 2–3 hr prior to inoculation. Use: For the cultivation of Clostridium perfringens. Lactose Gelatin Medium, Modified Composition per liter: Gelatin 120.0g Tryptose 15.0g Yeast extract 10.0g Lactose 10.0g Na 2 HPO 4 5.0g Phenol Red 0.05g pH 7.8 ± 0.2 at 25°C Source: This medium is available from HiMedia. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into screw-cap tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Just before use heat to remove oxygen and cool rapidly. Use: For the cultivation of Haemophilus spp. Lactose HiVeg Broth Composition per liter: Plant peptone 5.0g Lactose 5.0g Plant extract 3.0g pH 7.0 ± 0.2 at 25°C Source: This medium is available as a premixed powder from HiMe- dia. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes. Use: For the detection of coliform bacteria in water, foods, and dairy products as per standard methods. Lactose Lecithin Agar Composition per liter: Agar 15.0g Casein enzymic hydrolysate 12.65g Lactose 10.0g Peptic digest of animal tissue 5.5g NaCl 5.5g Yeast extract 3.85g Pancreatic digest of heart muscle 3.3g Corn starch 1.1g Egg lecithin 0.66g L-Cysteine·HCl·H 2 O 0.5g NaN 3 0.2g Neomycin sulfate 0.15g CaCl 2 ·2H 2 O 0.05g Bromcresol Purple 0.025g pH 6.8 ± 0.2 at 25°C Source: This medium is available from HiMedia. Caution: Sodium azide has a tendency to form explosive metal azides with plumbing materials. It is advisable to use enough water to flush off the disposables. 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. © 2010 by Taylor and Francis Group, LLC 930 Lactose Medium Use: For the isolation and differentiation of histotoxic clostridia from clinical specimens. Lactose Medium Composition per liter: Agar 15.0g K 2 HPO 4 0.5g (NH 4 ) 2 SO 4 0.5g NaCl 50.0mg Lactose solution 50.0mL pH 7.2 ± 0.2 at 25°C Lactose Solution: Composition per 50.0mL: Lactose 5.0g Preparation of Lactose Solution: Add lactose to distilled/deion- ized water and bring volume to 50.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: Add components, except lactose solu- tion, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Adjust pH to 7.2. Autoclave for 15 min at 15 psi pressure– 121°C. Aseptically add 50.0mL of sterile lactose solution. Mix thor- oughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation of unidentified bacterium ATCC 51468. Lactose Minimal Medium Composition per liter: Agar 20.0g Lactose 15.0g K 2 HPO 4 5.0g NH 4 Cl 2.0g NaCl 1.0g MgSO 4 0.1g Yeast extract 0.1g 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 Xanthomonas campestris. Lactose Peptone Agar Composition per liter: Agar 15.0g Lactose 0.5g Peptone 0.5g 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 Acytostelium subglobosum, numerous Dictyostelium species, Didymium nigripes, Drechslera bisep- tata, several Mortierella species, Nodulisporium griseobrunneum, Peni- cillium chrysogenum, Polysphondylium pallidum, Polysphondylium violaceum, Pythium insidiosum, and Trichomycete species. Lactose Peptone Agar, Double Strength Composition per liter: Agar 18.0g Lactose 1.0g Peptone 1.0g 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 various fungi. Lactose Peptone Agar, Half Strength Composition per liter: Agar 15.0g Lactose 0.25g Peptone 0.25g 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 various fungi. Lactose Peptone Broth Composition per liter: Casein enzymic hydrolysate 17.0g Lactose 10.0g NaCl 5.0g Papaic digest of soybean meal 3.0g Bromcresol Purple 0.02g pH 7.4 ± 0.2 at 25°C Source: This medium is available from HiMedia. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes with inverted Durham tubes. Autoclave for 15 min at 15 psi pressure– 121°C. Use: For the detection of coliform organisms in water. Lactose Ricinoleate Broth Composition per liter: Lactose 10.0g Peptone 5.0g Sodium ricinoleate 1.0g pH 7.6 ± 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 selective cultivation of members of the Enterobacteri- aceae. Lactose Sulfite Broth Base Composition per liter: Lactose 10.0g Casein enzymic hydrolysate 5.0g Yeast extract 2.5g NaCl 2.5g L-Cysteine·HCl·H 2 O 0.3g Sodium metabisulfite solution 50.0mL Ferric ammonium citrate 50.0mL pH 7.1 ± 0.2 at 25°C Source: This medium is available from HiMedia. © 2010 by Taylor and Francis Group, LLC Lasseur Medium 931 Selective Sodium Metabisulfite Solution: Composition per 100.0mL: Sodium metabisulfite 1.2g Preparation of Sodium Metabisulfite Solution: Add compo- nents to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Selective Ferric Ammonium Citrate Solution: Composition per 100.0mL: Ferric ammonium citrate 1.0g Preparation of Ferric Ammonium Citrate Solution: Add components to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except sodium meta- bisulfite and ferric ammonium citrate solutions, to distilled/deionized water and bring volume to 999.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Aseptically add sodium metabisulfite and ferric ammonium citrate solutions. Mix thoroughly. Pour into Petri dishes or aseptically distribute into sterile tubes. Use: For the detection and enumeration of Clostridium perfringens in pharmaceutical products. Lambda Broth Composition per liter: Pancreatic digest of casein 10.0g NaCl 2.5g Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 25 min at 15 psi pressure–121°C. Use: For the cultivation of Escherichia coli in the preparation of bac- teriophage lysates. Lambda Plates Composition per liter: Agar 10.0g Pancreatic digest of casein 10.0g NaCl 2.5g 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 in 45.0mL volumes per plate. Use: For use as a base agar to support the cultivation of Escherichia coli in the preparation of bacteriophages. Lambda Top Agar Composition per liter: Pancreatic digest of casein 10.0g Agar 7.0g NaCl 2.5g Preparation of Medium: 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°C. Distribute into flasks in 100.0mL volumes. Reautoclave for 15 min at 15 psi pressure–121°C. Store at 25°C. Use: For use as a top agar for the distribution of bacteriophage or Escherichia coli. Lange Medium Composition per liter: Agar 20.0g Maltose 5.0g Ca(NO 3 ) 2 0.5g MgSO 4 0.5g K 2 HPO 4 0.25g Peptone 0.1g Horse dung extract 100.0mL Horse Dung Extract: Composition per liter: Horse dung, fresh 100.0g Preparation of Horse Dung Extract: Add fresh horse dung to distilled/deionized water and bring volume to 1.0L. Autoclave for 50 min at 15 psi pressure–121°C. Filter through Whatman #1 filter paper. Reserve filtrate. 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 Coprinus xanthothrix. Lash Serum Medium Composition per liter: Casamino acids 14.0g NaCl 6.0g Glucose 2.0g Maltose 1.5g Sodium lactate (60% solution) 0.5g KCl 0.1g CaCl 2 ·2H 2 O 0.1g Serum solution 500.0mL pH 5.8 ± 0.2 at 25°C Serum Solution: Composition per 500.0mL: NaHCO 3 0.1g Bovine serum 200.0mL Preparation of Serum Solution: Add components to distilled/de- ionized water and bring volume to 500.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except serum solution, to distilled/deionized water and bring volume to 500.0mL. Mix thor- oughly. Distribute into tubes in 5.0mL volumes. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Aseptically add 5.0mL of ster- ile serum solution to each tube. Mix thoroughly. Use: For the cultivation of Trichomonas vaginalis from clinical spec- imens. Lasseur Medium (LMG 170) Composition per liter: Glycerol 25.0g Agar 15.0g L-Asparagine 9.0g MgSO 4 ·7H 2 O 5.0g K 2 HPO 4 2.5g CaCl 2 ·2H 2 O 0.54g FeSO 4 ·7H 2 O 0.1g pH 6.7 ± 0.2 at 25°C © 2010 by Taylor and Francis Group, LLC 932 Lauryl Sulfate Broth Preparation of Medium: Add K 2 HPO 4 to 20.0mL of distilled/de- ionized water. Mix thoroughly. Add remaining components. Bring vol- ume to 1.0L with distilled/deionized water. Gently heat and bring to boiling. Adjust pH to 6.7. 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 fluore- scens. Lauryl Sulfate Broth (m-Lauryl Sulfate Broth) Composition per liter: Peptone 39.0g Lactose 30.0g Yeast extract 6.0g Sodium lauryl sulfate 1.0g Phenol Red 0.2g 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 bottles or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation and enumeration of coliform bacteria, espe- cially Escherichia coli, in water by the membrane filter method. Lauryl Sulfate Broth (Lauryl Tryptose Broth) Composition per liter: Pancreatic digest of casein 20.0g Lactose 5.0g NaCl 5.0g K 2 HPO 4 2.75g KH 2 PO 4 2.75g Sodium lauryl sulfate 0.1g pH 6.8 ± 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 containing an inverted Durham tube in 10.0mL volumes. Autoclave for 12 min at 15 psi pressure–121°C. Cool broth quickly to 25°C. For test- ing water samples with 10.0mL volumes, prepare medium double strength. Use: For the detection of coliform bacteria in a variety of specimens. Also, for the enumeration of coliform bacteria by the multiple-tube fer- mentation technique. Lauryl Sulfate Broth, Fluorocult (Fluorocult Lauryl Sulfate Broth) Composition per liter: Tryptose 20.0g Lactose 5.0g NaCl 5.0g K 2 HPO 4 2.75g KH 2 PO 4 2.75g L-tryptophan 1.g Sodium lauryl sulfate 0.1g 4-Methylumbelliferyl-ß-D-glucuronide 0.1g pH 6.8 ± 0.2 at 25°C Source: This medium is available from Merck. Preparation of Medium: Add components to distilled/deionized wa- ter and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Cool. Distribute into test tubes containing inverted Durham tubes. Autoclave for 15 min at 15 psi pressure–121°C. The prepared broth is clear and yellowish-brown. Use: For the detection of E. coli in milk. The medium complies with the German-DIN-Norm 10183 for the examination of milk, with the regulations acc. to § 35 LMBG (01.00/54) for the examination of food, and according to ISO/DIS 11886-2.2 (1994) for milk and milk prod- ucts. The lauryl sulfate largely inhibits the growth of undesirable microbial flora. The presence of E. coli is indicated by fluorescence under a long wavelength UV lamp. A positive indole reaction and gas formation due to fermentation of lactose confirm the results. Lauryl Sulfate Broth with MUG Composition per liter: Pancreatic digest of casein 20.0g Lactose 5.0g NaCl 5.0g K 2 HPO 4 2.75g KH 2 PO 4 2.75g Sodium lauryl sulfate 0.1g 4-Methylumbelliferyl-β-D-glucuronide (MUG) 0.05g pH 6.8 ± 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 containing an inverted Durham tube in 10.0mL volumes. Autoclave for 12 min at 15 psi pressure–121°C. Cool broth quickly to 25°C. For test- ing water samples with 10.0mL volumes, prepare medium double strength. Use: For the detection of Escherichia coli in water and food samples by a fluorogenic procedure. Lauryl Sulfate HiVeg Broth (Lauryl Tryptose HiVeg Broth) Composition per liter: Plant hydrolysate No. 1 20.0g NaCl 5.0g Lactose 5.0g K 2 HPO 4 2.75g KH 2 PO 4 2.75g Sodium lauryl sulfate 0.1g pH 6.8 ± 0.2 at 25°C Source: This medium is available as a premixed powder from HiMe- dia. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into bottles or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation and enumeration of coliform bacteria in water, wastewater, dairy products, and other foods. © 2010 by Taylor and Francis Group, LLC LB Broth, Modified 933 Lauryl Tryptose Broth See: Lauryl Sulfate Broth Lauryl Tryptose Broth with MUG (Lauryl Sulfate Broth with MUG) (LST-MUG) (BAM M77) Composition per liter: Pancreatic digest of casein 20.0g Lactose 5.0g NaCl 5.0g K 2 HPO 4 2.75g KH 2 PO 4 2.75g Sodium lauryl sulfate 0.1g 4-Methylumbelliferyl-β-D-glucuronide (MUG) 0.05g pH 6.8 ± 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 containing an inverted Durham tube in 10.0mL volumes. Autoclave for 12 min at 15 psi pressure–121°C. Cool broth quickly to 25°C. For test- ing water samples with 10.0mL volumes, prepare medium double strength. Use: For the detection of Escherichia coli in water and food samples by a fluorogenic procedure. Lauryl Tryptose Mannitol Broth with Tryptophan Composition per liter: Pancreatic digest of casein 20.0g Lactose 5.0g NaCl 5.0g K 2 HPO 4 2.75g KH 2 PO 4 2.75g Sodium lauryl sulfate 0.1g L-Tryptophan 0.2g pH 6.8 ± 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 wa- ter and bring volume to 1.0L. Mix thoroughly. Distribute into tubes con- taining an inverted Durham tube in 10.0mL volumes. Autoclave for 10 min at 10 psi pressure–115°C. Cool broth quickly to 25°C. Use: For the detection of Escherichia coli in water samples. LAVMm2 Medium Composition per liter: Lactalbumin hydrolysate 10.0g Sodium acetate 5.0g MgCl 2 ·6H 2 O 20.3mg Nitrilotriacetic acid 19.1mg CaCl 2 11.1mg FeSO 4 0.152mg Thiamine·HCl 0.05mg Cupric acetate 0.04mg Biotin 0.02mg pH 8.0–8.1 at 25°C Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.5 with Na 2 CO 3 . Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. The pH should be 8.0–8.1 after autoclaving. Use: For the cultivation of Caryophanon latum. LB Agar Composition per liter: Agar 15.0g Pancreatic digest of casein 10.0g NaCl 5.0g Yeast extract 5.0g 1N NaOH 1.0mL 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. Adjust pH to 7.0. Distribute into tubes or flasks. Autoclave for 25 min at 15 psi pressure–121°C. Pour into sterile Petri dishes in 35–40.0mL volumes. Use: For the cultivation of Escherichia coli. LB Agar See: Lactobacillus bulgaricus Agar LB Broth, Modified Composition per liter: Pancreatic digest of casein 10.0g NaCl 5.8g Yeast extract 5.0g NaCl solution 16.8mL Glucose solution 10.0mL CaCl 2 ·2H 2 O solution 2.0mL MgCl 2 solution 1.6mL pH 7.0 ± 0.2 at 25°C NaCl Solution: Composition per 100.0mL: NaCl 25.0g Preparation of NaCl Solution: Add NaCl to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Glucose Solution: Composition per 100.0mL: D-Glucose 40.0g Preparation of Glucose Solution: Add D-glucose to distilled/de- ionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. CaCl 2 ·2H 2 O Solution: Composition per 10.0mL: CaCl 2 ·2H 2 O 0.735g Preparation of CaCl 2 ·2H 2 O Solution: Add CaCl 2 ·2H 2 O to dis- tilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. MgCl 2 Solution: Composition per 10.0mL: MgCl 2 0.95g Preparation of MgCl 2 Solution: Add MgCl 2 to distilled/deion- ized water and bring volume to 10.0mL. Mix thoroughly. Filter steril- ize. © 2010 by Taylor and Francis Group, LLC 934 LB Medium Preparation of Medium: Add components—except NaCl solution, glucose solution, CaCl 2 ·2H 2 O solution, and MgCl 2 solution—to dis- tilled/deionized water and bring volume to 969.6mL. Mix thoroughly. Adjust pH to 7.0. Autoclave for 30 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add 16.8mL of sterile NaCl solution, 10.0mL of sterile glucose solution, 2.0mL of sterile CaCl 2 ·2H 2 O solution, and 1.6mL of sterile MgCl 2 solution. Mix thoroughly. Aseptically distrib- ute into sterile tubes or flasks. Use: For the cultivation and maintenance of Escherichia coli. LB Medium (LB Broth, Miller) (ATCC Medium 1065) (ATCC Medium 1082) Composition per liter: NaCl 10.0g Pancreatic digest of casein 10.0g Yeast extract 5.0g Source: This medium is available from BD Diagnostic 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 maintenance of Bacillus subtilis, Coryne- bacterium glutamicum, Enterobacter cloacae, Erwinia uredovora, Escherichia coli, Klebsiella oxytoca, and Salmonella choleraesuis. LB Medium (Luria-Bertani Medium) Composition per liter: NaCl 10.0g Pancreatic digest of casein 10.0g Yeast extract 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. Adjust pH to 7.0. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure– 121°C. Use: For the cultivation of Bacillus subtilis, Daptobacter species, and Escherichia coli. LB Medium (Luria Broth) (Lenox Broth) Composition per liter: Pancreatic digest of casein 10.0g NaCl 5.0g Yeast extract 5.0g pH 7.0 ± 0.2 at 25°C Preparation of Medium: Add components to distilled/deionized wa- ter and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.0. Distribute into tubes or flasks. Autoclave for 25 min at 15 psi pressure–121°C. Use: For the cultivation of Escherichia coli. LB Medium with Ampicillin (ATCC Medium 1315) Composition per liter: NaCl 10.0g Pancreatic digest of casein 10.0g Yeast extract 5.0g Ampicillin solution 10.0mL pH 7.0 ± 0.2 at 25°C Ampicillin Solution: Composition per 10.0mL: Ampicillin 0.1mg Preparation of Ampicillin Solution: Add ampicillin to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except ampicillin solu- tion, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Adjust pH to 7.0. Autoclave for 15 min at 15 psi pressure– 121°C. Cool to 45°–50°C. Aseptically add sterile ampicillin solution. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation and maintenance of Escherichia coli. LB Medium with Ampicillin (ATCC Medium 1364) Composition per liter: NaCl 10.0g Pancreatic digest of casein 10.0g Yeast extract 5.0g Ampicillin solution 10.0mL pH 7.0 ± 0.2 at 25°C Ampicillin Solution: Composition per 10.0mL: Ampicillin 0.02mg Preparation of Ampicillin Solution: Add ampicillin to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except ampicillin solu- tion, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Adjust pH to 7.0. Autoclave for 15 min at 15 psi pressure– 121°C. Cool to 45°–50°C. Aseptically add sterile ampicillin solution. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation and maintenance of Escherichia coli. LB Medium with Chloramphenicol Composition per liter: NaCl 10.0g Pancreatic digest of casein 10.0g Yeast extract 5.0g Chloramphenicol 0.01g 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. Dis- tribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure– 121°C. Use: For the cultivation and maintenance of Escherichia coli. LB Medium with Glucose Composition per liter: NaCl 10.0g Pancreatic digest of casein 10.0g Glucose 5.0g Yeast extract 5.0g pH 7.0 ± 0.2 at 25°C © 2010 by Taylor and Francis Group, LLC . ster- ilize skim milk. To 1.0L of cooled, sterile agar mixture, aseptically add 150.0mL of sterile skim milk, 36.0mL of sterile egg yolk emulsion, 50%, and 20.0mL of sterile inhibitor solution cultivation of Escherichia coli in the preparation of bac- teriophage lysates. Lambda Plates Composition per liter: Agar 10.0g Pancreatic digest of casein 10.0g NaCl 2.5g Preparation of Medium:. the cultivation of Escherichia coli in the preparation of bacteriophages. Lambda Top Agar Composition per liter: Pancreatic digest of casein 10.0g Agar 7.0g NaCl 2.5g Preparation of Medium: Add