Handbook of Microbiological Media, Fourth Edition part 94 pot

5.0g Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L.. Preparation of Medium: Add components to distilled/deionized water and bring volume to

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Lactobacillus rimae Medium 925

Na-acetate·3H2O 5.0g

Glucose 4.0g

Maltose 4.0g

Meat extract 3.0g

K2HPO4·3H2O 2.6g

(NH4)2 citrate 2.0g

Cysteine-HCl·H2O 0.5g

MgSO4·7H2O 0.1g

MnSO4·4H2O 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

Compositionper liter:

Maltose 20.0g

Peptone 10.0g

Yeast extract 10.0g

Glucose 5.0g

Use: For the cultivation of Lactobacillus brevis.

Lactobacillus MRS HiVeg Agar

(MRS HiVeg Agar) Compositionper 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

K2HPO4 2.0g

Polysorbate 80 1.0g

MgSO4 0.1g

MnSO4 0.05g

pH 6.5 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from

HiMe-dia

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) Compositionper liter:

Glucose 20.0g

Plant extract 10.0g

Plant peptone No 3 10.0g

Sodium acetate 5.0g

Yeast extract 5.0g

K2HPO4 2.0g Polysorbate 80 1.0g MgSO4 0.1g MnSO4 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

Milk, peptonized 15.0g Glucose 10.0g Yeast extract 5.0g

KH2PO4 2.0g Sorbitan monooleate complex 1.0g Orotic acid 25.0mg

D-Pantothenic acid 0.2mg Tomato juice 100.0mL

L-Cysteine·HCl·H2O solution 7.5mL

pH 6.8 ± 0.2 at 25°C

L-Cysteine·HCl·H2O Solution:

Compositionper 10.0mL:

L-Cysteine·HCl·H2O 0.15g

Preparation of L-Cysteine·HCl·H2O Solution: Add L -cyste-ine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL Mix thoroughly

to boiling Continue boiling for 2–3 min Distribute into bottles in 20.0mL volumes Add 0.15mL of L-cysteine·HCl·H2O 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

Yeast extract 10.0g Peptone 5.0g Pancreatic digest of casein 5.0g Glucose 5.0g (NH4)2SO4 0.5g

L-Cysteine·HCl 0.5g Resazurin 1.0mg Mineral solution 40.0mL Fatty acid mixture 3.1mL Hemin solution 0.5mL Vitamin K1 0.2mL

pH 6.9 ± 0.2 at 25°C

Mineral Solution:

NaHCO3 10.0g NaCl 2.0g

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926 Lactobacillus rimae Medium with Tween™

KH2PO4 1.0g

K2HPO4 1.0g

MgSO4·7H2O 0.48g

CaCl2·2H2O 0.3g

Preparation of Mineral Solution: Add components to distilled/

deionized water and bring volume to 1.0L Mix thoroughly

Fatty Acid Mixture:

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:

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% CO2 Add L-cysteine·HCl, hemin solution, and fatty acid mixture

Adjust pH to 6.9 with 8N NaOH while continuing to sparge with 100%

CO2 After pH has been reached, sparge with 100% N2 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™

Yeast extract 10.0g

Peptone 5.0g

Pancreatic digest of casein 5.0g

Glucose 5.0g

(NH4)2SO4 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 K1 0.2mL

pH 6.9 ± 0.2 at 25°C

Mineral Solution:

NaHCO3 10.0g

NaCl 2.0g

KH2PO4 1.0g

K2HPO4 1.0g

MgSO4·7H2O 0.48g

CaCl2·2H2O 0.3g

Preparation of Mineral Solution: Add components to distilled/ deionized water and bring volume to 1.0L Mix thoroughly

Fatty Acid Mixture:

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:

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% CO2 Add L-cysteine·HCl, hemin solution, and fatty

acid mixture Adjust pH to 6.9 with 8N NaOH while continuing to

sparge with 100% CO2 After pH has been reached, sparge with 100%

N2 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

KH2PO4 6.0g Yeast extract 5.0g Ammonium citrate 2.0g Polysorbate 80 1.0g MgSO4·7H2O 0.575g MnSO4·2H2O 0.12g FeSO3 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

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Lactococcus piscium Medium 927

Lactobacillus Selection Broth

See: LBS™ Broth

Lactobacillus Selection HiVeg Agar Base

with Acetic Acid and Polysorbate

Sodium acetate·3H2O 25.0g

Glucose 20.0g

Agar 15.0g

Plant hydrolysate 10.0g

KH2PO4 6.0g

Yeast extract 5.0g

MgSO4 0.575g

FeSO4 0.034g

MnSO4 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

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

Sodium acetate 25.0g

Glucose 20.0g

KH2PO4 6.0g

Yeast extract 5.0g

Polysorbate 80 1.0g

MgSO4 0.575g

MnSO4 0.12g

FeSO4 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

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·3H2O 25.0g

Glucose 20.0g

Agar 15.0g

KH2PO4 6.0g Yeast extract 5.0g Ammonium citrate 2.0g Oxgall 1.5g Polysorbate 80 1.0g MgSO4 0.575g MnSO4 0.12g FeSO4 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

KH2PO4 6.0g Yeast extract 5.0g Ammonium citrate 2.0g Oxgall 1.5g Polysorbate 80 1.0g MgSO4·7H2O 0.575g MnSO4·2H2O 0.12g FeSO4·7H2O 0.034g Acetic acid, glacial 1.32mL

pH 5.4 ± 0.2 at 25°C

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

Glucose 10.0g Peptone 10.0g Beef extract 8.0g NaCl 5.0g Yeast extract 3.0g

KH2PO4 1.5g

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928 Lactose Blue Agar

MgSO4·7H2O 0.2g

MnSO4·4H2O 0.05g

pH 6.8 ± 0.2 at 25°C

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)

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

HiMe-dia

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)

Lactose 15.5g

Agar 13.0g

NaCl 5.0g

Plant extract 3.5g

Bromthymol Blue 0.04g

pH 7.0 ± 0.2 at 25°C

HiMe-dia

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 Compositionper 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 Compositionper liter:

Lactose 37.5g Agar 15.0g Casein hydrolysate 3.0g

KH2PO4 1.0g MnSO4 0.5g Microelements solution 2.0mL

pH 6.0 ± 0.2 at 25°C

Microelements Solution:

Fe(NO3)3·9H2O 723.5mg ZnSO4·7H2O 439.8mg MnSO4·4H2O 203.0mg

H2SO4 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 Compositionper liter:

Lactose 20.0g Distillers solubles 15.0g Yeast, autolyzed 5.0g

pH 7.0 ± 0.2 at 25°C

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 Compositionper 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

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Lactose Lecithin Agar 929

Columbia Blood Agar Base:

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:

Neomycin sulfate 0.18g

NaN3 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

NaN3 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 Compositionper 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

Na2HPO4 5.0g Phenol Red 0.05g

pH 7.8 ± 0.2 at 25°C

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 Compositionper liter:

Plant peptone 5.0g Lactose 5.0g Plant extract 3.0g

pH 7.0 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0mL 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·H2O 0.5g NaN3 0.2g Neomycin sulfate 0.15g CaCl2·2H2O 0.05g Bromcresol Purple 0.025g

pH 6.8 ± 0.2 at 25°C

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

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930 Lactose Medium

Use: For the isolation and differentiation of histotoxic clostridia from

clinical specimens

Lactose Medium Compositionper liter:

Agar 15.0g

K2HPO4 0.5g

(NH4)2SO4 0.5g

NaCl 50.0mg

Lactose solution 50.0mL

pH 7.2 ± 0.2 at 25°C

Lactose Solution:

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

Agar 20.0g

Lactose 15.0g

K2HPO4 5.0g

NH4Cl 2.0g

NaCl 1.0g

MgSO4 0.1g

Yeast extract 0.1g

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 Compositionper liter:

Agar 15.0g

Lactose 0.5g

Peptone 0.5g

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

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 Compositionper 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

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 Compositionper 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

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·H2O 0.3g Sodium metabisulfite solution 50.0mL Ferric ammonium citrate 50.0mL

pH 7.1 ± 0.2 at 25°C

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Lasseur Medium 931

Selective Sodium Metabisulfite Solution:

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:

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 Compositionper liter:

NaCl 2.5g

Use: For the cultivation of Escherichia coli in the preparation of

bac-teriophage lysates

Lambda Plates Compositionper liter:

Agar 10.0g

NaCl 2.5g

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 Compositionper liter:

Agar 7.0g

NaCl 2.5g

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 Compositionper liter:

Agar 20.0g Maltose 5.0g Ca(NO3)2 0.5g MgSO4 0.5g

K2HPO4 0.25g Peptone 0.1g Horse dung extract 100.0mL

Horse Dung Extract:

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

Use: For the cultivation and maintenance of Coprinus xanthothrix.

Lash Serum Medium Compositionper liter:

Casamino acids 14.0g NaCl 6.0g Glucose 2.0g Maltose 1.5g Sodium lactate (60% solution) 0.5g KCl 0.1g CaCl2·2H2O 0.1g Serum solution 500.0mL

pH 5.8 ± 0.2 at 25°C

Serum Solution:

NaHCO3 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) Compositionper liter:

Glycerol 25.0g Agar 15.0g

L-Asparagine 9.0g MgSO4·7H2O 5.0g

K2HPO4 2.5g CaCl2·2H2O 0.54g FeSO4·7H2O 0.1g

pH 6.7 ± 0.2 at 25°C

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932 Lauryl Sulfate Broth

Preparation of Medium: Add K2HPO4 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

water and bring volume to 1.0L Mix thoroughly Distribute into bottles

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) Compositionper liter:

Lactose 5.0g

NaCl 5.0g

K2HPO4 2.75g

KH2PO4 2.75g

pH 6.8 ± 0.2 at 25°C

Di-agnostic Systems

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)

Tryptose 20.0g

Lactose 5.0g

NaCl 5.0g

K2HPO4 2.75g

KH2PO4 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

K2HPO4 2.75g

KH2PO4 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) Compositionper liter:

Plant hydrolysate No 1 20.0g NaCl 5.0g Lactose 5.0g

K2HPO4 2.75g

KH2PO4 2.75g Sodium lauryl sulfate 0.1g

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 bottles

Use: For the cultivation and enumeration of coliform bacteria in water, wastewater, dairy products, and other foods

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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:

Lactose 5.0g

NaCl 5.0g

K2HPO4 2.75g

KH2PO4 2.75g

4-Methylumbelliferyl-β-D-glucuronide (MUG) 0.05g

pH 6.8 ± 0.2 at 25°C

Di-agnostic Systems

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:

Lactose 5.0g

NaCl 5.0g

K2HPO4 2.75g

KH2PO4 2.75g

L-Tryptophan 0.2g

pH 6.8 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Oxoid

Unipath

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 Compositionper liter:

Lactalbumin hydrolysate 10.0g

Sodium acetate 5.0g

MgCl2·6H2O 20.3mg

Nitrilotriacetic acid 19.1mg

CaCl2 11.1mg

FeSO4 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

Na2CO3 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 Compositionper 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

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 CaCl2·2H2O solution 2.0mL MgCl2 solution 1.6mL

pH 7.0 ± 0.2 at 25°C

NaCl Solution:

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:

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:

CaCl2·2H2O 0.735g

Preparation of CaCl 2 ·2H 2 O Solution: Add CaCl2·2H2O to dis-tilled/deionized water and bring volume to 10.0mL Mix thoroughly Filter sterilize

MgCl 2 Solution:

MgCl2 0.95g

Preparation of MgCl 2 Solution: Add MgCl2 to distilled/deion-ized water and bring volume to 10.0mL Mix thoroughly Filter steril-ize

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934 LB Medium

Preparation of Medium: Add components—except NaCl solution,

glucose solution, CaCl2·2H2O solution, and MgCl2 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 CaCl2·2H2O solution, and

1.6mL of sterile MgCl2 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) Compositionper liter:

NaCl 10.0g

Yeast extract 5.0g

Source: This medium is available from BD Diagnostic Systems

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) Compositionper liter:

NaCl 10.0g

Yeast extract 5.0g

pH 7.0 ± 0.2 at 25°C

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) Compositionper liter:

NaCl 5.0g

Yeast extract 5.0g

pH 7.0 ± 0.2 at 25°C

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) Compositionper liter:

NaCl 10.0g

Yeast extract 5.0g Ampicillin solution 10.0mL

pH 7.0 ± 0.2 at 25°C

Ampicillin Solution:

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) Compositionper 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:

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

LB Medium with Chloramphenicol Compositionper 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

LB Medium with Glucose Compositionper 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

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