Handbook of Microbiological Media, Fourth Edition part 93 doc

1.0g Preparation of Listeria mono Enrichment Supplement II So-lution: Add L-phosphatidylinositol to distilled/deionized water and bring volume to 10.0mL.. 76,700U Preparation of Listeri

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L mono Differential HiVeg Agar Base 915

Listeria mono Enrichment Supplement II Solution:

Compositionper 10.0mL:

L-phosphatidylinositol 1.0g

Preparation of Listeria mono Enrichment Supplement II

So-lution: Add L-phosphatidylinositol to distilled/deionized water and

bring volume to 10.0mL Mix thoroughly Filter sterilize

Listeria mono Selective Supplement I Solution:

Polymyxin B sulfate 76,700U

Preparation of Listeria mono Selective Supplement I

Solu-tion: Add polymyxin B sulfate to distilled/deionized water and bring

volume to 10.0mL Mix thoroughly Filter sterilize

Listeria mono Selective Supplement II Solution:

Ceftazidime 20.0mg

Nalidixic acid, sodium salt 20.0mg

Amphotericin B 10.0mg

Preparation of Listeria mono Selective Supplement II

Solu-tion: Add components to distilled/deionized water and bring volume

to 10.0mL Mix thoroughly Filter sterilize

Preparation of Medium: Add components, except Listeria mono

enrichment supplement II, Listeria mono selective supplement I, and

Listeria mono selective supplement II, to distilled/deionized water and

bring volume to 970.0mL Mix thoroughly Gently heat and bring to

boiling Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–

50°C Aseptically add 10.0mL Listeria mono enrichment supplement

II, 10.0mL Listeria mono selective supplement I, and 10.0mL Listeria

mono selective supplement II Mix thoroughly Pour into sterile Petri

dishes or distribute into sterile tubes

Use: For the selective and differential isolation of Listeria

monocyto-genes from clinical and food specimens.

L mono Confirmatory HiVeg Agar Base

Compositionper liter:

Plant special peptone 30.0g

Agar 12.0g

LiCl 10.0g

B.C indicator 8.6g

Yeast extract 6.0g

NaCl 5.0g

α-Methyl-D-mannoside 3.0g

Na2HPO4, anhydrous 2.5g

Listeria mono enrichment supplement II 10.0mL

Listeria mono selective supplement I 10.0mL

Listeria mono selective supplement II 10.0mL

pH 7.2 ± 0.2 at 25°C

Source: This medium, without Listeria mono enrichment supplement

II, Listeria mono selective supplement I, and Listeria mono selective

supplement II, is available as a premixed powder from HiMedia

Caution: LiCl is harmful Avoid bodily contact and inhalation of

va-pors On contact with skin wash with plenty of water immediately

So-lution: Add L-phosphatidylinositol to distilled/deionized water and

bring volume to 10.0mL Mix thoroughly Filter sterilize

Solu-tion: Add polymyxin B sulfate to distilled/deionized water and bring volume to 10.0mL Mix thoroughly Filter sterilize

Ceftazidime 20.0mg Nalidixic acid, sodium salt 20.0mg Amphotericin B 10.0mg

Preparation of Medium: Add components, except Listeria mono enrichment supplement II, Listeria mono selective supplement I, and

bring volume to 970.0mL Mix thoroughly Gently heat and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–

L mono Differential HiVeg Agar Base

Plant peptone No 1 18.0g Agar 15.0g LiCl 10.0g Yeast extract 10.0g Plant hydrolysate 6.0g NaCl 5.0g

Na2HPO4, anhydrous 2.5g Glucose 2.0g Sodium pyruvate 2.0g Magnesium glycerophosphate 1.0g MgSO4 0.5g Chromogenic substrate 0.05g

Listeria mono enrichment supplement II 10.0mL Listeria mono selective supplement I 10.0mL Listeria mono selective supplement II 10.0mL

pH 7.2 ± 0.2 at 25°C

Source: This medium, without Listeria mono enrichment supplement

II, Listeria mono selective supplement I, and Listeria mono selective

supplement II, is available as a premixed powder from HiMedia

Caution: LiC lis harmful Avoid bodily contact and inhalation of va-pors On contact with skin wash with plenty of water immediately

So-lution: Add L-phosphatidylinositol to distilled/deionized water and bring volume to 10.0mL Mix thoroughly Filter sterilize

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916 Lab-Lemco Agar

Solu-tion: Add polymyxin B sulfate to distilled/deionized water and bring

volume to 10.0mL Mix thoroughly Filter sterilize

Ceftazidime 20.0mg

Nalidixic acid, sodium salt 20.0mg

Amphotericin B 10.0mg

Preparation of Medium: Add components, except Listeria mono

enrichment supplement II, Listeria mono selective supplement I, and

bring volume to 970.0mL Mix thoroughly Gently heat and bring to

boiling Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–

L Salts for Thermophiles

See: Mineral Salts for Thermophiles

Lab-Lemco Agar Compositionper liter:

Agar 15.0g

Peptone 5.0g

Lab Llemco meat 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 and bring

to boiling Distribute into tubes or flasks Autoclave for 15 min at 15

psi pressure–121°C Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation and maintenance of a variety of heterotrophic

microorganisms

Lab-Lemco Broth Compositionper liter:

Peptone 5.0g

Lab Lemco meat extract 3.0g

pH 7.4 ± 0.2 at 25°C

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

or flasks Autoclave for 15 min at 15 psi pressure–121°C

Use: For the cultivation of a variety of heterotrophic microorganisms,

including microorganisms from wastewater

Lachica’s Medium

See: SA Agar, Modified

Lachica’s Medium Base Composition per liter:

Beef heart, infusion from 500.0g Agar 15.0g Tryptose 10.0g NaCl 5.0g Amylose Azure 3.0g Selective supplement solution 10.0mL

pH 7.4 ± 0.2 at 25°C

Source: This medium is available from HiMedia

Selective Supplement Solution:

Ampicillin 10.0mg

Preparation of Selective Supplement Solution: Add ampicillin

to distilled/deionized water and bring volume to 10.0mL Mix thor-oughly Filter sterilize

Preparation of Medium: Add components, except selective sup-plement solution, to distilled/deionized water and bring volume to 990.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure– 121°C Cool to 50°C Aseptically add selective supplement solution Mix thoroughly Pour into Petri dishes or aseptically distribute into sterile tubes

Use: For the isolation and cultivation of Aeromonas hydrophila from

foods stored under different temperature conditions

Lactate Agar Compositionper liter:

Yeast extract 3.0g

K2HPO4 2.8g Agar 2.0g Peptone 2.0g

KH2PO4 0.52g Sodium lactate (60% solution) 10.0mL

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Gently heat and bring

to boiling Adjust pH to 7.2 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 Serpens flexibilis.

Lactate Broth Compositionper liter:

Yeast extract 3.0g

K2HPO4 2.8g Peptone 2.0g

KH2PO4 0.52g Sodium lactate (60% solution) 10.0mL

pH 7.2 ± 0.2 at 25°C

to boiling Adjust pH to 7.2 Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C

Use: For the cultivation and maintenance of Serpens flexibilis.

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Lactic Agar 917

Lactate Seawater Minimal Medium

Composition per liter:

Tris(hydroxymethyl)aminomethane·HCl 7.88g

Sodium or potassium lactate 2.0g

NH4Cl 1.0g

K2HPO4·3H2O 0.075g

FeSO4·7H2O 0.028g

Artificial seawater 500.0mL

pH 7.5 ± 0.2 at 25°C

Artificial Seawater:

MgSO4·7H2O 24.7g

NaCl 23.4g

CaCl2·2H2O 2.9g

KCl 1.5g

Preparation of Artificial Seawater: Add components to distilled/

deionized water and bring volume to 1.0L Mix thoroughly

water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.5

Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–

121°C

Use: For the cultivation of ATCC strains 27134 and 27136

Lactic Acid Bacteria Broth

Sodium acetate 12.0g

Pancreatic digest of casein 10.0g

Yeast autolysate 5.0g

Glucose 10.0g

Solution A 5.0mL

Solution B 5.0mL

pH 5.1 ± 0.2 at 25°C

Source: Yeast autolysate is available from Oxoid Unipath

Solution A:

K2HPO4 10.0g

KH2PO4 10.0g

Preparation of Solution A: Add components to distilled/deionized

water and bring volume to 100.0mL Mix thoroughly

Solution B:

MgSO4·7H2O 4.0g

FeSO4·7H2O 0.2g

MnSO4·H2O 0.2g

NaCl 0.2g

Preparation of Solution B: Add components to distilled/deionized

water and bring volume to 100.0mL Mix thoroughly

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

Use: For the cultivation of Lactobacillus malefermentans.

Lactic Acid Bacteria Medium

Agar 15.0g

Glucose 10.0g

KH2PO4 5.0g

Peptone 5.0g

Sodium acetate·3H2O 5.0g Yeast extract 5.0g Diammonium hydrogen citrate 2.0g Tween™ 80 1.0g Sorbic acid 0.5g MgSO4·7H2O 0.5g MnSO4·4H2O 0.2g FeSO4·7H2O 0.05g

pH 5.3–5.4 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 5.3–5.4 Gently heat and bring to boiling Distribute into tubes or flasks Auto-clave for 15 min at 15 psi pressure–121°C Pour into sterile Petri dishes

or leave in tubes

Use: For the isolation and cultivation of lactic acid bacteria from wine

Lactic Acid Bacteria Selective Agar Base Composition per liter:

Casein enzymic hydrolysate 20.0g Agar 17.0g Maltose 10.0g Yeast extract 5.0g Fructose 5.0g Glucose 5.0g Potassium aspartate 2.5g Potassium glutamate 2.5g Betaine hydrochloride 2.0g Diammonium hydrogen citrate 2.0g MgSO4·7H2O 2.0g

KH2PO4 2.0g Liver concentrate 1.0g MnSO4·2H2O 0.66g

N-acetyl glucosamine 0.5g

Selective supplement solution 20.0mL

pH 5.4 ± 0.2 at 25°C

Selective Supplement Solution:

2-Phenylethanol 3.0g Cycloheximide 10.0mg Polysorbate 80 10.0mL

Preparation of Selective Supplement Solution: Add compo-nents to distilled/deionized water and bring volume to 20.0mL Mix thoroughly Filter sterilize

Caution: Cycloheximide is toxic Avoid skin contact or aerosol for-mation and inhalation

Preparation of Medium: Add components, except selective sup-plement solution, to distilled/deionized water and bring volume to 980.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure– 121°C Cool to 50°C Aseptically add selective supplement solution Mix thoroughly Pour into Petri dishes or aseptically distribute into sterile tubes

Use: For the selective isolation of lactic acid bacteria from beer and brewing processes

Lactic Agar Composition per liter:

Casein enzymatic hydrolysate 20.0g Agar 15.0g

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918 Lactic Agar for Yogurt Bacteria, Modified

Yeast extract 5.0g

Glucose 5.0g

Lactose 5.0g

Sucrose 5.0g

NaCl 4.0g

Gelatin 2.5g

Sodium acetate 1.5g

Ascorbic acid 0.5g

Use: For the enumeration and identification of lactic streptococci and

lactobacilli by the pour plate method

Lactic Agar for Yogurt Bacteria, Modified

Elliker agar 1.0L

Milk solution 70.0mL

pH 6.8 ± 0.1 at 25°C

Elliker Agar:

Agar 15.0g

Yeast extract 10.0g

Gelatin 4.0g

Glucose 3.0g

Ascorbic acid 2.5g

Lactose 2.5g

NaCl 2.5g

Sodium acetate 2.5g

Sucrose 2.5g

Preparation of Elliker Agar: Add components to

distilled/deion-ized 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 45°–50°C

Milk Solution:

Nonfat dry milk solids 11.0g

Preparation of Milk Solution: Add nonfat dry milk solids to

dis-tilled/deionized water and bring volume to 100.0mL Mix thoroughly

Autoclave for 12 min at 15 psi pressure–121°C Cool to 45°–50°C

Preparation of Medium: Add 70.0mL of sterile milk solution to

1.0L of cooled, sterile Elliker agar Mix thoroughly Pour into sterile

Petri dishes Allow plates to dry at 28°–30°C for 18–24 hr

Use: For the cultivation of acidogenic microorganisms, especially

Lactobacillus species and lactic streptococci, from foods.

Lactic Bacteria Broth Compositionper liter:

Glucose 10.0g

Yeast autolysate 5.0g

Solution A 5.0mL

Solution B 5.0mL

pH 5.1–5.3 at 25°C

Solution A:

K2HPO4 10.0g

KH2PO4 10.0g

Preparation of Solution A: Add components to distilled/deionized water and bring volume to 100.0mL Mix thoroughly

Solution B:

MgSO4·7H2O 4.0g FeSO4 0.2g MnSO4·5H2O 0.2g NaCl 0.2g

Preparation of Solution B: Add components to distilled/deionized water and bring volume to 100.0mL Mix thoroughly

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Distribute into tubes

Use: For the cultivation and maintenance of Lactobacillus buchneri,

Lactobacillus delbrueckii, and Pediococcus damnosus.

Lactic Bacteria Differential Agar Composition per liter:

Agar 15.0g Casein enzymic hydrolysate 10.0g Casein acid hydrolysate 3.0g Fructose 2.5g

KH2PO4 2.5g Papaic digest of soybean meal 1.5g Yeast extract 1.0g Polysorbate 80 1.0g Bromcresol Green 0.055g

pH 7.0 ± 0.2 at 25°C

Source: This medium, without polysorbate 80, is available from Hi-Media

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 differentiation of homofermentative and heterofermenta-tive lactic acid bacteria

Lactic Bacteria Differential HiVeg Agar Compositionper liter:

Agar 15.0g Plant hydrolysate 10.0g Plant acid hydrolysate 3.0g Fructose 2.5g

KH2PO4 2.5g Papaic digest of soybean meal 1.5g Yeast extract 1.0g Bromcresol Green 0.055g Polysorbate 80 1.0mL

pH 7.0 ± 0.2 at 25°C

Source: This medium, without polysorbate 80, is available as a pre-mixed powder from HiMedia

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 5.3–5.4

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Lactobacilli Agar, AOAC 919

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 differentiation of homofermentative and

heterofermenta-tive lactic acid bacteria

Lactic Bacteria Differential HiVeg Broth

Plant hydrolysate 10.0g

Plant acid hydrolysate 3.0g

Fructose 2.5g

KH2PO4 2.5g

Papaic digest of soybean meal 1.5g

Yeast extract 1.0g

Bromcresol Green 0.055g

Polysorbate 80 1.0mL

pH 7.0 ± 0.2 at 25°C

Source: This medium, without polysorbate 80, is available as a

pre-mixed powder from HiMedia

water and bring volume to 1.0L Mix thoroughly Adjust pH to 5.3–5.4

Auto-clave for 15 min at 15 psi pressure–121°C

Use: For the differentiation of homofermentative and

heterofermenta-tive lactic acid bacteria

Lactic HiVeg Agar Compositionper liter:

Agar 15.0g

Glucose 5.0g

Lactose 5.0g

Sucrose 5.0g

Yeast extract 5.0g

NaCl 4.0g

Sodium acetate 1.5g

Ascorbic acid 0.5g

pH 6.0 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from

HiMe-dia

water and bring volume to 1.0L Mix thoroughly Adjust pH to 5.3–5.4

Auto-clave for 15 min at 15 psi pressure–121°C

Use: For the enumeration and identification of lactic streptococci and

lactobacilli by the pour plate method

Lactic Streak Agar Compositionper liter:

Agar 15.0g

Sodium carboxymethylcellulose 10.0g

Calcium citrate 10.0g

Beef extract 5.0g

Papaic digest of soybean meal 5.0g

Polypeptone™ 5.0g

Yeast extract 5.0g

Lactose 1.5g

L-Arginine·HCl 1.5g Bromcresol Purple (0.1% solution) 20.0mL

pH 6.0 ± 0.1 at 25°C

Preparation of Medium: Add components—except Bromcresol Purple solution, calcium citrate, and sodium carboxymethylcellu-lose—to distilled/deionized water and bring volume to 800.0mL Mix thoroughly Gently heat and bring to boiling In a blender, add the cal-cium citrate and sodium carboxymethylcellulose to 200.0mL of dis-tilled/deionized water Blend until mixed Add the 200.0mL of citrate/ carboxymethylcellulose solution to the hot agar solution Mix thor-oughly Adjust pH to 6.0 Distribute into flasks in 100.0mL volumes Autoclave for 10 min at 10 psi pressure–115°C Cool to 45°–50°C Aseptically add 2.0mL of sterile Bromcresol Purple to each flask Mix thoroughly Pour into sterile Petri dishes Allow plates to dry at 37°C for 1 hr before use

Use: For the differentiation of lactic streptococci Bacteria that pro-duce acid from lactose appear as yellow colonies

Lactic Streak HiVeg Agar Compositionper liter:

Agar 15.0g Calcium citrate 10.0g Sodium carboxymethylcellulose 10.0g Plant extract 5.0g Plant peptone 5.0g Papaic digest of soybean meal 5.0g Yeast extract 5.0g

L-Arginine hydrochloride 1.5g Lactose 1.5g Bromcresol Purple 2.0mg

pH 6.0 ± 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 Adjust pH to 6.0 Autoclave for 15 min at 15 psi pres-sure–121°C Pour into sterile Petri dishes

Use: For the differentiation of lactic streptococci Bacteria that pro-duce acid from lactose appear as yellow colonies

Lactobacilli Agar, AOAC (Lactobacilli Agar, Association of Official Analytical Chemists) Compositionper liter:

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

KH2PO4 2.0g Sorbitan monooleate complex 1.0g Tomato juice 100.0mL

pH 6.8 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from BD Di-agnostic Systems

to boiling Continue boiling for 2–3 min Distribute into tubes in

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920 Lactobacilli Broth, AOAC

10.0mL volumes Autoclave for 15 min at 15 psi pressure–121°C

Al-low tubes to cool in an upright position

Use: For the cultivation and maintenance of stock cultures of

Lacto-bacillus casei ATCC 7469, LactoLacto-bacillus fermentum ATCC 9338,

Lactobacillus leichmannii ATCC 4797, and Lactobacillus viridescens

ATCC 12706 used in the microbiological assay of vitamins

Lactobacilli Broth, AOAC

(Lactobacilli Broth, Association of

Official Analytical Chemists)

Milk, peptonized 15.0g

Glucose 10.0g

Yeast extract 5.0g

KH2PO4 2.0g

Sorbitan monooleate complex 1.0g

Tomato juice 100.0mL

pH 6.8 ± 0.2 at 25°C

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

Di-agnostic Systems

to boiling Continue boiling for 2–3 min Distribute into tubes in

10.0mL volumes Autoclave for 15 min at 15 psi pressure–121°C

Use: For the cultivation and preparation of inocula of stock cultures of

Lactobacillus casei ATCC 7469, Lactobacillus fermentum ATCC

9338, Lactobacillus leichmannii ATCC 4797, and Lactobacillus

viri-descens ATCC 12706 used in the microbiological assay of vitamins.

Lactobacilli deMan-Rogosa-Sharpe Broth

See:Lactobacilli MRS Broth

Lactobacilli HiVeg Broth (Elliker Broth, HiVeg) Compositionper liter:

Plant extract 20.0g

Yeast extract 10.0g

Gelatin 4.0g

Glucose 3.0g

Ascorbic acid 2.5g

Lactose 2.5g

NaCl 2.5g

Sodium acetate 2.5g

Sucrose 2.5g

pH 6.8 ± 0.1 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 Autoclave for 15 min at 15 psi pressure–121°C

Use: For the cultivation of Lactobacillus species and lactic

strepto-cocci, from foods.

Lactobacilli MRS Broth (Lactobacilli deMan-Rogosa-Sharpe Broth)

Glucose 20.0g

Beef extract 10.0g

Peptone 10.0g Sodium acetate 5.0g Yeast extract 5.0g Ammonium citrate 2.0g

Na2HPO4 2.0g Tween™ 80 1.0g MgSO4·7H2O 0.1g MnSO4·5H2O 0.05g

pH 6.5 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from BD Di-agnostic Systems

to boiling Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C

Use: For the cultivation of Lactobacillus species Also used for the cultivation and maintenance of Aerococcus viridans, Bifidobacterium

coryneforme, Lactococcus plantarum, Leuconostoc species, Pectina-tus cerevisiiphilus, Pediococcus species, and Sporolactobacillus inuli-nus.

Lactobacilli MRS Broth with Mevalonic Acid

See: Pediococcus Medium

with Mevalonic Acid Lactobacilli MRS Broth

See: Pediococcus Medium

Lactobacilli MRS Broth with Cysteine Compositionper liter:

Glucose 20.0g Beef extract 10.0g Peptone 10.0g Sodium acetate 5.0g Yeast extract 5.0g

L-Cysteine 2.0g Ammonium citrate 2.0g

Na2HPO4 2.0g Tween™ 80 1.0g MgSO4·7H2O 0.1g MnSO4·5H2O 0.05g

pH 6.5 ± 0.2 at 25°C

to boiling Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C

Use: For the cultivation and maintenance of Lactobacillus species, including L ruminis and L vitulinus

Lactobacilli MRS Broth with 0.5% Cysteine

(ATCC 1844) Compositionper liter:

Glucose 20.0g Beef extract 10.0g Peptone 10.0g Sodium acetate 5.0g Yeast extract 5.0g

L-Cysteine·HCl 5.0g Ammonium citrate 2.0g

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Lactobacillus bulgaricus Agar 921

Na2HPO4 2.0g

Tween™ 80 1.0g

MgSO4·7H2O 0.1g

MnSO4·5H2O 0.05g

pH 6.5 ± 0.2 at 25°C

psi pressure–121°C

Use: For the cultivation of Lactobacillus sharpeae.

Lactobacilli MRS Broth with Ethanol

Glucose 20.0g

Beef extract 10.0g

Peptone 10.0g

Sodium acetate 5.0g

Yeast extract 5.0g

L-Cysteine 2.0g

Ammonium citrate 2.0g

Na2HPO4 2.0g

Tween™ 80 1.0g

MgSO4·7H2O 0.1g

MnSO4·5H2O 0.05g

Ethanol (95% solution) 100.0mL

pH5.0 ± 0.2 at 25°C

Preparation of Medium: Add components, except ethanol, to

dis-tilled/deionized water and bring volume to 990.0mL Mix thoroughly

Gently heat and bring to boiling Autoclave for 15 min at 15 psi

pres-sure–121°C Cool to 25°C Filter sterilize ethanol Aseptically add

100.0mL of sterile ethanol Mix thoroughly Aseptically distribute into

sterile tubes or flasks

Use: For the cultivation and maintenance of Lactobacillus species.

Lactobacillus Agar 2

Glucose 20.0g

Agar 15.0g

Tryptic digest of casein 10.0g

Yeast extract 5.0g

Meat extract 2.0g

K2HPO4 0.5g

KH2PO4 0.5g

MgSO4·7H2O 200.0mg

DL-Mevalonic acid 30.0mg

FeSO4·7H2O 10.0mg

MnSO4·H2O 7.5mg

Ethanol 40.0mL

Tween™ 80 1.0mL

pH 5.2 ± 0.2 at 25°C

Use: For the cultivation and maintenance of Lactobacillus

fructiv-orans and Lactobacillus homohiochii.

Lactobacillus bifidus Medium

Lactose 70.0g Sodium acetate, anhydrous 50.0g Pancreatic digest of casein 10.0g

K2HPO4 5.0g Tween™ 80 1.0g Alanine 0.4g

L-Cystine 0.4g Tryptophan 0.4g Asparagine 0.2g Adenine 0.02g Guanine 0.02g Uracil 0.02g Xanthine 0.02g Pyridoxine·HCl 2.4mg Nicotinic acid 1.2mg Calcium pantothenate 0.8mg Riboflavin 0.4mg Thiamine·HCl 0.4mg

p-Aminobenzoic acid 0.02mg

Folic acid 0.02mg Biotin 8.0μg Ascorbic acid solution 100.0mL Human milk, skimmed 20.0mL Salts B 10.0mL

pH 6.8 ± 0.2 at 25°C

Salts B:

MgSO4·7H2O 10.0g FeSO4·7H2O 0.5g NaCl 0.5g MnSO4·2H2O 0.337g

Preparation of Salts B: Add components to distilled/deionized wa-ter and bring volume to 250.0mL Mix thoroughly

Ascorbic Acid Solution:

Ascorbic acid 1.0g

Preparation of Ascorbic Acid Solution: Add ascorbic acid to distilled/deionized water and bring volume to 100.0mL Mix

thorough-ly Adjust pH to 6.5 Filter sterilize

Preparation of Medium: Add components, except ascorbic acid solution and human milk, to distilled/deionized water and bring vol-ume to 900.0mL Mix thoroughly Gently heat and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C Aseptically add 100.0mL of sterile ascorbic acid solution Mix thor-oughly This constitutes a double-strength medium To prepare a sin-gle-strength medium, aseptically combine 500.0mL of sterile double-strength medium, 480.0mL of sterile distilled/deionized water, and 20.0mL of sterile human milk Aseptically distribute into sterile tubes

or flasks

Use: For the cultivation and maintenance of Bifidobacterium

(Lacto-bacillus) bifidum.

Lactobacillus bulgaricus Agar

(LB Agar) Compositionper 900.0mL:

Agar 20.0g Glucose 20.0g

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922 Lactobacillus bulgaricus HiVeg Agar Base with Tomato Juice and Acetate Buffer

Beef extract 10.0g

Yeast extract 5.0g

K2HPO4 2.0g

Tween™ 80 1.0g

Acetate buffer 80.0mL

Tomato juice, filtered 40.0mL

pH 6.8 ± 0.2 at 25°C

Acetate Buffer:

Acetic acid 10.0mL

Preparation of Acetate Buffer: Add components to

distilled/de-ionized water and bring volume to 1.0L Mix thoroughly

Preparation of Medium: Add components, except acetate buffer,

to distilled/deionized water and bring volume to 920.0mL Mix

thor-oughly Gently heat and bring to boiling Adjust pH to 6.8 Add

80.0mL of acetate buffer Mix thoroughly Autoclave for 15 min at 15

psi pressure–121°C Pour into sterile Petri dishes

Use: For the isolation, cultivation, and enumeration of Lactobacillus

bulgaricus from foods.

Lactobacillus bulgaricus HiVeg Agar Base

with Tomato Juice and Acetate Buffer

Agar 20.0g

Glucose 20.0g

Plant extract 10.0g

Yeast extract 5.0g

K2HPO4 2.0g

Tomato juice 2.0g

Polysorbate 80 1.0g

Acetate buffer 80.0mL

Tomato juice, filtered 40.0mL

pH 6.8 ± 0.2 at 25°C

Source: This medium, without tomato juice and acetate buffer, is

available as a premixed powder from HiMedia

Acetate Buffer:

Acetic acid 10.0mL

Preparation of Acetate Buffer: Add components to

distilled/de-ionized water and bring volume to 1.0L Mix thoroughly

Preparation of Medium: Add components, except acetate buffer,

to distilled/deionized water and bring volume to 920.0mL Mix

thor-oughly Gently heat and bring to boiling Adjust pH to 6.8 Add

80.0mL of acetate buffer Mix thoroughly Autoclave for 15 min at 15

psi pressure–121°C Pour into sterile Petri dishes

Use: For the isolation, cultivation, and enumeration of Lactobacillus

bulgaricus from foods.

Lactobacillus Chloramphenicol Agar 1

Milk, peptonized 15.0g

Agar 10.0g

Glucose 10.0g

Yeast extract 5.0g

KH2PO4 2.0g Sorbitan monooleate complex 1.0g Chloramphenicol 0.3g Tomato juice 100.0mL

pH 6.8 ± 0.2 at 25°C

to boiling Continue boiling for 2–3 min Distribute into tubes in 10.0mL volumes Autoclave for 15 min at 15 psi pressure–121°C Al-low tubes to cool in an upright position

Use: For the cultivation of Lactobacillus casei.

Lactobacillus Chloramphenicol Broth 1

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

KH2PO4 2.0g Sorbitan monooleate complex 1.0g Chloramphenicol 0.3g Tomato juice 100.0mL

pH 6.8 ± 0.2 at 25°C

to boiling Continue boiling for 2–3 min Distribute into tubes in 10.0mL volumes Autoclave for 15 min at 15 psi pressure–121°C

Use: For the cultivation of Lactobacillus casei.

Lactobacillus Chloramphenicol Medium 2

Glucose 18.5g Agar 13.5g Pancreatic digest of gelatin 10.0g Beef extract 8.0g Yeast extract 4.0g Sodium acetate 3.0g

K2HPO4 2.0g Ammonium citrate 2.0g Polysorbate 80 1.0g MgSO4·7H2O 0.2g Chloramphenicol 0.1g MnSO4·4H2O 0.05g

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 while stirring and bring to boiling Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation of Lactobacillus casei and Pediococcus

acidi-lactici.

Lactobacillus Heteroferm Screen Agar

(MRS, Modified) Compositionper liter:

Glucose 20.0g Agar 15.0g Proteose peptone No 3 10.0g

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Lactobacillus Medium 923

Sodium acetate 5.0g

Yeast extract 5.0g

2-Phenylethyl alcohol 3.0g

K2HPO4 2.0g

MgSO4·7H2O 0.1g

MnSO4·H2O 0.05g

Cycloheximide 4.0mg

Tween™ 80 1.0mL

pH 5.5 ± 0.01 at 25°C

Caution: Cycloheximide is toxic Avoid skin contact or aerosol

for-mation and inhalation

water and bring volume to 1.0L Mix thoroughly Autoclave for 15 min

at 15 psi pressure–121°C Cool to 50°C Adjust pH to 5.5 with glacial

acetic acid Pour into sterile Petri dishes

Use: For the isolation and cultivation of Lactobacillus species from

salad dressings

Lactobacillus Heteroferm Screen Broth

(MRS, Modified) Compositionper liter:

Glucose 20.0g

Proteose peptone No 3 10.0g

Sodium acetate 5.0g

Yeast extract 5.0g

2-Phenylethyl alcohol 3.0g

K2HPO4 2.0g

MgSO4·7H2O 0.1g

MnSO4·H2O 0.05g

Cycloheximide 4.0mg

Tween™ 80 1.0mL

pH 4.3 ± 0.01 at 25°C

Caution: Cycloheximide is toxic Avoid skin contact or aerosol

for-mation and inhalation

water and bring volume to 1.0L Mix thoroughly Adjust pH to 4.3 with

concentrated HCl Distribute into test tubes that contain an inverted

Durham tube Autoclave for 15 min at 15 psi pressure–121°C

Use: For the isolation and cultivation of Lactobacillus species from

foods

Lactobacillus homohiochii Medium

D-Glucose 20.0g

Tryptic digest of casein 10.0g

Yeast extract 5.0g

Meat extract 2.0g

K2HPO4 0.5g

KH2PO4 0.5g

MgSO4·7H2O 0.2g

Sodium acetate 20.0mg

FeSO4·7H2O 10.0mg

MnSO4·H2O 7.5mg

Ethanol 40.0mL 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 Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure– 121°C

homo-hiochii.

Lactobacillus Medium

(ATCC Medium 78) Compositionper liter:

Pancreatic digest of casein 20.0g Agar 15.0g Tryptose 5.0g Yeast extract 5.0g Glucose 3.0g Lactose 2.0g Liver extract concentrate 1.0g Tween™ 80 0.05g Tomato juice, filtered 200.0mL

pH 6.5 ± 0.2 at 25°C

Use: For the cultivation and maintenance of Lactobacillus fermentum and Lactobacillus salivarius.

Glucose 10.0g Proteose peptone 7.5g Yeast extract 7.5g

KH2PO4 2.0g L-Cysteine·HCl·H2O 1.0g Tween™ 80 0.1g Tomato juice, filtered 100.0mL

pH 7.0 ± 0.2 at 25°C

Use: For the cultivation and maintenance of Lactobacillus casei.

Agar 20.0g Pancreatic digest of casein 10.0g Glucose 5.0g Yeast extract 5.0g

KH2PO4 3.0g

K2HPO4 3.0g

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924 Lactobacillus Medium

Tryptose 3.0g

Sodium acetate 1.0g

L-Cysteine·HCl·H2O 0.2g

Salt solution R 5.0mL

Tween™ 80 1.0mL

pH 6.3 ± 0.2 at 25°C

Salt Solution R:

MgSO4·7H2O 11.5g

MnSO4·2H2O 2.4g

FeSO4·7H2O 0.68g

Preparation of Salt Solution R: Add components to

distilled/de-ionized water and bring volume to 100.0mL Mix thoroughly Store at

4°C

Use: For the cultivation and maintenance of Lactobacillus delbrueckii

and Lactobacillus jensenii.

Glucose 20.0g

Tryptone 10.0g

Yeast extract 5.0g

Beef extract 2.0g

K2HPO4 0.5g

KH2PO4 0.5g

MgSO4·7H2O 200.0mg

FeSO4·7H2O 10.0mg

MnSO4·H2O 7.5mg

Ethanol 40.0mL

Mevalonic acid solution 10.0mL

Tween™ 80 1.0mL

pH 5.2 ± 0.2 at 25°C

Mevalonic Acid Solution:

Preparation of Mevalonic Acid Solution: Add DL-mevalonic

acid to distilled/deionized water and bring volume to 10.0mL Mix

thoroughly Filter sterilize

Preparation of Medium: Add components, except mevalonic acid

solution and ethanol, to distilled/deionized water and bring volume to

950.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–

121°C Aseptically add 40.0mL of filter-sterilized ethanol and 10.0mL

of sterile mevalonic acid solution Mix thoroughly Aseptically

distrib-ute into sterile tubes or flasks

Use: For the cultivation of Lactobacillus homohiochii.

Lactobacillus Medium (Lactobacillus Sake Medium)

Agar 13.0g

Yeast extract 5.0g

Liver extract concentrate 0.2g

Sake 700.0mL

fructiv-orans and Lactobacillus homohiochii.

Lactobacillus Medium II

(DSMZ Medium 93) Compositionper liter:

Glucose 20.0g Na-acetate 20.0g Agar 15.0g Pancreatic digest of casein 10.0g Yeast extract 5.0g Meat extract 2.0g

KH2PO4 0.5g

K2HPO4 0.5g MgSO4·7H2O 0.2g

DL-Mevalonic acid 30.0mg FeSO4·7H2O 10.0mg MnSO4·2H2O 7.5mg Ethanol 40.0mL Tween™ 80 1.0mL

pH 5.2 ± 0.2 at 25°C

Use: For the cultivation and maintenance of Lactobacillus fructivorans and Lactobacillus homohiochii.

Lactobacillus Medium III

Pancreatic digest of casein 10.0g Fructose 7.0g Glucose 7.0g Maltose 7.0g Meat extract 5.0g Sodium acetate·3H2O 5.0g Yeast extract 5.0g

K2HPO4·3H2O 2.6g Diammonium citrate 2.0g Sodium gluconate 2.0g

L-Cysteine·HCl 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 Adjust pH to 6.3 Dis-tribute into tubes or flasks Autoclave for 15 min at 15 psi pressure– 121°C

Use: For the cultivation of Lactobacillus species.

Lactobacillus Medium IV

(DSMZ Medium 859) Compositionper liter:

Tryptone 10.0g Yeast extract 5.0g

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