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B

Authored by: Ronald M. Atlas

Handbook of Microbiological Media

Print publication date:  March  2010
Online publication date:  March  2010

Print ISBN: 9781439804063
eBook ISBN: 9781439804087
Adobe ISBN:

10.1201/EBK1439804063-c4

 

Abstract

B Broth

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B

B Broth

(Medium for Ureaplasma )

Composition per 100.25mL:

Yeast extract

0.1g

GHL (Glycyl-L–histidyl-L–lysine)

2.0μg

PPLO broth without Crystal Violet

50.0mL

Horse serum, not inactivated

10.0mL

Bromthymol Blue (0.4% solution)

1.0mL

Urea solution

0.25mL

pH 6.0 ± 0.2 at 25°C

B Broth

(Medium for Ureaplasma )

Composition per 100.25mL:

Yeast extract

0.1g

GHL (Glycyl-L–histidyl-L–lysine)

2.0μg

PPLO broth without Crystal Violet

50.0mL

Horse serum, not inactivated

10.0mL

Bromthymol Blue (0.4% solution)

1.0mL

Urea solution

0.25mL

pH 6.0 ± 0.2 at 25°C

PPLO Broth without Crystal Violet:

Composition per 50.0mL:

Beef heart, infusion from

1.62g

Peptone

0.32g

NaCl

0.16g

Source: PPLO broth without Crystal Violet is available as a premixed powder from BD Diagnostic Systems.

Preparation of PPLO Broth without Crystal Violet: Add components to distilled/deionized water and bring volume to 50.0mL. Mix thoroughly.

Urea Solution:

Composition per 10.0mL:

Urea

1.0g

Preparation of Urea Solution: Add urea to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components—except GHL, urea solution, and horse serum—to double glass-distilled water and bring volume to 90.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. To 90.0mL of the sterile medium, aseptically add 2.0μg of GHL, 10.0mL of horse serum, and 0.25mL of sterile urea solution. Mix thoroughly. Aseptically distribute into tubes or flasks.

Use: For the cultivation and maintenance of Ureaplasma urealyticum and other Ureaplasma species.

B/1t 7 A Medium

Composition per liter:

Agar

20.0g

K2HPO4

7.0g

KH2PO4

3.0g

Glucose

2.0g

(NH4)2SO4

1.0g

MgSO4·7H2O

0.1g

CaCl2·2H2O

0.01g

Indole

0.01g

FeSO4·7H2O

0.5mg

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 Escherichia coli and other bacteria.

B12 Assay HiVeg Medium

(Vitamin B12 Assay HiVeg Medium)

Composition per liter:

Glucose

40.0g

Sodium acetate

20.0g

Plant hydrolysate

15.0g

Ascorbic acid

4.0g

Polysorbate 80

2.0g

K2HPO4

1.0g

KH2PO4

1.0g

DL-Tryptophan

0.4g

MgSO4·7H2O

0.4g

L-Cysteine

0.4g

Asparagine

0.2g

Adenine sulfate

0.02g

FeSO4

0.02g

Guanine hydrochloride

0.02g

MnSO4

0.02g

NaCl

0.02g

Uracil

0.02g

Xanthine

0.02g

Pyridoxal·HCl

4.0mg

Pyridoxine·HCl

4.0mg

Niacin

2.0mg

p-Aminobenzoic acid

2.0mg

Riboflavin

1.0mg

Thymine·HCl

1.0mg

Calcium pantothenate

1.0mg

Pyridoxamine·HCl

0.8mg

Folic acid

0.2mg

Biotin

0.01mg

pH 6.0 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Hi-Media.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the determination of the vitamin B12 content of pharmaceutical products and other materials. Lactobacillus leischmanii ATCC 7830 is used as a test organism. A standard curve can be generated by adding known concentrations of cyanocobalamin and measuring the growth response turbidimetrically at 530 nm.

B12 Assay Medium

Composition per liter:

Glucose

20.5g

Lactose

20.0g

Amino acids, vitamin-free casamino acids

15.0g

Sodium acetate

10.0g

K2HPO4

2.5g

Polysorbate 80

2.0g

Ascorbic acid

1.0g

L-Arginine

0.5g

L-Histidine

0.25g

L-Phenylalanine

0.25g

L-Valine

0.25g

L-Asparagine

0.2g

MgSO4·7H2O

0.2g

Mercaptoacetic acid

0.13g

Calcium pantothenate

0.1g

L-Tryptophan

0.1g

MnSO4

0.08g

Adenine

0.04g

Guanine

0.04g

Thymine

0.04g

Uracil

0.04g

(NH4)2SO4·FeSO4·6H2O

0.03g

KCN

5.0mg

Pyridoxal·HCl

1.0mg

Niacin

1.0mg

Riboflavin

1.0mg

Thiamine·HCl

0.5mg

p-Aminobenzoic acid

0.5mg

Folic acid

0.05mg

pH 6.0 ± 0.2 at 25°C

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

Caution: Cyanide is toxic.

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. Allow precipitate to settle out. Distribute supernatant into tubes in 5.0mL volumes. Add standard solution or test solutions to each tube. Adjust the volume of each tube to 10.0mL with distilled/deionized water. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the determination of the vitamin B12 content of pharmaceutical products and other materials. Lactobacillus leischmanii ATCC 7830 is used as a test organism. A standard curve can be generated by adding known concentrations of cyanocobalamin and measuring the growth response turbidimetrically at 530 nm.

B12 Culture Agar, USP

Composition per liter:

Agar

15.0g

Glucose

10.0g

Proteose peptone No. 3

7.5g

Yeast extract

7.5g

KH2PO4

2.0g

Polysorbate 80

0.1g

Tomato juice

100.0mL

pH 6.8 ± 0.1 at 25°C

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

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes in 10.0mL volumes. Autoclave for 15 min at 15 psi pressure–121°C. Cool tubes in an upright position.

Use: For the cultivation and maintenance of Lactobacillus leischmannii ATCC 7830 to be used as the test organism in the Vitamin B12 assay according to the USP.

B12 Inoculum Broth, USP

Composition per liter:

Glucose

10.0g

Proteose peptone No. 3

7.5g

Yeast extract

7.5g

K2HPO4

2.0g

Polysorbate 80

0.1g

Tomato juice

100.0mL

pH 6.8 ± 0.1 at 25°C

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

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes in 10.0mL volumes. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the preparation of inoculum cultures of Lactobacillus leischmanii ATCC 7830, which is used as the test organism in the vitamin B12 assay according to the USP.

B12 Medium

See: Vitamin B12 Medium

B12 Medium

(DSMZ Medium 236)

Composition per liter:

Agar

15.0g

Casein hydrolysate

6.0g

K2HPO4

0.2g

MgSO4·7H2O

0.2g

Asparagine

0.15g

Vitamin B12

40.0µg

FeSO4·7H2O

trace

Glycerol

2.0mL

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. 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 Escherichia coli.

B12 Nutrient Agar

See: Vitamin B12 Nutrient Agar

BA Medium

See: BA Medium with Cellulose

BA Medium with Cellobiose

Composition per liter:

NaHCO3

2.6g

NH4Cl

1.0g

Yeast extract

0.75g

K2HPO4·3H2O

0.4g

MgCl2·6H2O

0.1g

NaCl

0.1g

CaCl2·2H2O

0.05g

Resazurin

0.5mg

Cellobiose solution

50.0mL

Na2S·9H2O solution

10.0mL

Wolfe’s mineral solution

10.0mL

Wolfe’s vitamin solution

10.0mL

pH 6.9–7.0 at 25°C

Cellobiose Solution:

Composition per 50.0mL:

Cellobiose

4.0g

Preparation of Cellobiose Solution: Add cellobiose to distilled/ deionized water and bring volume to 50.0mL. Mix thoroughly. Filter sterilize.

Na2S·9H2O Solution:

Composition per 10.0mL:

Na2S·9H2O

0.25g

Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N2. Autoclave for 15 min at 15 psi pressure–121°C. Before use, neutralize to pH 7.0 with sterile HCl.

Wolfe’s Mineral Solution:

Composition per liter:

MgSO4·7H2O

3.0g

Nitrilotriacetic acid

1.5g

NaCl

1.0g

MnSO4·2H2O

0.5g

CoCl2·6H2O

0.1g

ZnSO4·7H2O

0.1g

CaCl2·2H2O

0.1g

FeSO4·7H2O

0.1g

NiCl2·6H2O

0.025g

KAl(SO4)2·12H2O

0.02g

CuSO4·5H2O

0.01g

H3BO3

0.01g

Na2MoO4·2H2O

0.01g

Na2SeO3·5H2O

0.3mg

Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with KOH. Add remaining components. Add distilled/deionized water to 1.0L. Adjust pH to 6.8.

Wolfe’s Vitamin Solution:

Composition per liter:

Pyridoxine·HCl

10.0mg

p-Aminobenzoic acid

5.0mg

Lipoic acid

5.0mg

Nicotinic acid

5.0mg

Riboflavin

5.0mg

Thiamine·HCl

5.0mg

Calcium DL-pantothenate

5.0mg

Biotin

2.0mg

Folic acid

2.0mg

Vitamin B12

0.1mg

Preparation of Wolfe’s Vitamin Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize.

Preparation of Medium: Prepare and dispense medium under 80% N2 + 20% CO2 gas mixture. Add components, except cellobiose solution, Na2S·9H2O solution, Wolfe’s mineral solution, and Wolfe’s vita-min solution, to distilled/deionized water and bring volume to 920.0mL. Mix thoroughly. Sparge with 80% N2 + 20% CO2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add 50.0mL of sterile cellobiose solution, 10.0mL of sterile Wolfe’s mineral solutionn, 10.0mL of sterile Wolfe’s vitamin solution, and 10.0mL of sterile Na2S·9H2O solution. Mix thoroughly. Aseptically and anaerobically distribute into sterile tubes or bottles.

Use: For the cultivation of Caldicellulosiruptor lactoaceticus.

BA Medium with Cellulose

(DSMZ Medium 671)

Composition per liter:

NaHCO3

2.6g

Cellulose

2.0g

NH4Cl

1.0g

Yeast extract

0.75g

K2HPO4·3H2O

0.4g

MgCl2·6H2O

0.1g

NaCl

0.1g

CaCl2·2H2O

0.05g

Resazurin

0.5mg

Na2S·9H2O solution

10.0mL

Wolfe’s mineral solution

10.0mL

Wolfe’s vitamin solution

10.0mL

pH 6.9–7.0 at 25°C

Na2S·9H2O Solution:

Composition per 10.0mL:

Na2S·9H2O

0.25g

Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N2. Autoclave for 15 min at 15 psi pressure–121°C. Before use, neutralize to pH 7.0 with sterile HCl.

Wolfe’s Mineral Solution:

Composition per liter:

MgSO4·7H2O

3.0g

Nitrilotriacetic acid

1.5g

NaCl

1.0g

MnSO4·2H2O

0.5g

CoCl2·6H2O

0.1g

ZnSO4·7H2O

0.1g

CaCl2·2H2O

0.1g

FeSO4·7H2O

0.1g

NiCl2·6H2O

0.025g

KAl(SO4)2·12H2O

0.02g

CuSO4·5H2O

0.01g

H3BO3

0.01g

Na2MoO4·2H2O

0.01g

Na2SeO3·5H2O

0.3mg

Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with KOH. Add remaining components. Add distilled/deionized water to 1.0L. Adjust pH to 6.8.

Wolfe’s Vitamin Solution:

Composition per liter:

Pyridoxine·HCl

10.0mg

p-Aminobenzoic acid

5.0mg

Lipoic acid

5.0mg

Nicotinic acid

5.0mg

Riboflavin

5.0mg

Thiamine·HCl

5.0mg

Calcium DL-pantothenate

5.0mg

Biotin

2.0mg

Folic acid

2.0mg

Vitamin B12

0.1mg

Preparation of Wolfe’s Vitamin Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize.

Preparation of Medium: Prepare and dispense medium under 80% N2 + 20% CO2 gas mixture. Add components, except Na2S·9H2O solution, Wolfe’s mineral solution, and Wolfe’s vitamin solution, to distilled/deionized water and bring volume to 970.0mL. Mix thoroughly. Sparge with 80% N2 + 20% CO2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add 10.0mL of sterile Wolfe’s mineral solution, 10.0mL of sterile Wolfe’s vitamin solution, and 10.0mL of sterile Na2S·9H2O solution. Mix thoroughly. Aseptically and anaerobically distribute into sterile tubes or bottles.

Use: For the cultivation of Caldicellulosiruptor lactoaceticus and Caldicellulosiruptor kristjanssonii.

Baar’s Medium for Sulfate Reducers

Composition per liter:

Sodium lactate

3.5g

MgSO4·7H2O

2.0g

K2HPO4

1.0g

CaSO4

1.0g

NH4Cl

0.5g

Ferrous ammonium sulfate solution

10.0mL

Yeast extract solution

10.0mL

pH 7.5 ± 0.2 at 25°C

Ferrous Ammonium Sulfate Solution:

Composition per 10.0mL:

Fe(NH4)2(SO4)2

0.5g

Preparation of Ferrous Ammonium Sulfate Solution: Add Fe(NH4)2(SO4)2 to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C.

Yeast Extract Solution:

Composition per 10.0mL:

Yeast extract

1.0g

Preparation of Yeast Extract Solution: Add yeast extract to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C.

Preparation of Medium: Add components, except ferrous ammonium sulfate solution and yeast extract solution, to tap water and bring volume to 980.0mL. 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 of sterile ferrous ammonium sulfate solution and sterile yeast extract solution. Aseptically distribute into tubes or flasks.

Use: For the cultivation and maintenance of Desulfotomaculum nigrifi-cans.

Baar’s Medium for Sulfate Reducers, Modified

Composition per 1020.0mL:

Component I

400.0mL

Component III

400.0mL

Component II

200.0mL

Ferrous ammonium sulfate solution

20.0mL

pH 7.5 ± 0.2 at 25°C

Component I:

Composition per 400.0mL:

Sodium citrate

5.0g

MgSO4

2.0g

CaSO4

1.0g

NH4Cl

1.0g

Preparation of Component I: Add components to distilled/deionized water and bring volume to 400.0mL. Mix thoroughly. Adjust pH to 7.5. Autoclave for 15 min at 15 psi pressure–121°C.

Component II:

Composition per 200.0mL:

K2HPO4

0.5g

Preparation of Component II: Add K2HPO4 to distilled/deionized water and bring volume to 200.0mL. Mix thoroughly. Adjust pH to 7.5. Autoclave for 15 min at 15 psi pressure–121°C.

Component III:

Composition per 400.0mL:

Sodium lactate

3.5g

Yeast extract

1.0g

Preparation of Component III: Add components to distilled/deionized water and bring volume to 400.0mL. Mix thoroughly. Adjust pH to 7.5. Autoclave for 15 min at 15 psi pressure–121°C.

Ferrous Ammonium Sulfate Solution:

Composition per 20.0mL:

Fe(NH4)2(SO4)2

1.0g

Preparation of Ferrous Ammonium Sulfate Solution: Add Fe(NH4)2(SO4)2 to distilled/deionized water and bring volume to 20.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Aseptically combine component I, component II, and component III. Mix thoroughly. Distribute 5.0mL volumes into tubes under 97% N2 + 3% H2. Add medium to tubes while still warm to exclude as much O2 as possible. Aseptically add 0.1mL of sterile ferrous ammonium sulfate solution to 5.0mL of medium immediately prior to inoculation.

Use: For the cultivation and maintenance of Desulfovibrio, Desulfobulbus, Desulfotomaculum, and Thermodesulfobacterium species.

Baar’s Medium for Sulfate Reducers,

Modified with 2.5% Sodium Chloride

Composition per 1020.0mL:

Component I

400.0mL

Component III

400.0mL

Component II

200.0mL

Ferrous ammonium sulfate solution

20.0mL

pH 7.5 ± 0.2 at 25°C

Component I:

Composition per 400.0mL:

NaCl

25.0g

Sodium citrate

5.0g

MgSO4

2.0g

CaSO4

1.0g

NH4Cl

1.0g

Preparation of Component I: Add components to distilled/deionized water and bring volume to 400.0mL. Mix thoroughly. Adjust pH to 7.5. Autoclave for 15 min at 15 psi pressure–121°C.

Component II:

Composition per 200.0mL:

K2HPO4

0.5g

Preparation of Component II: Add K2HPO4 to distilled/deionized water and bring volume to 200.0mL. Mix thoroughly. Adjust pH to 7.5. Autoclave for 15 min at 15 psi pressure–121°C.

Component III:

Composition per 400.0mL:

Sodium lactate

3.5g

Yeast extract

1.0g

Preparation of Component III: Add components to distilled/deionized water and bring volume to 400.0mL. Mix thoroughly. Adjust pH to 7.5. Autoclave for 15 min at 15 psi pressure–121°C.

Ferrous Ammonium Sulfate Solution:

Composition per 20.0mL:

Fe(NH4)2(SO4)2

1.0g

Preparation of Ferrous Ammonium Sulfate Solution: Add Fe(NH4)2(SO4)2 to distilled/deionized water and bring volume to 20.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Aseptically combine component I, component II, and component III. Mix thoroughly. Distribute 5.0mL volumes into tubes under 97% N2 + 3% H2. Add medium to tubes while still warm to exclude as much O2 as possible. Aseptically add 0.1mL of sterile ferrous ammonium sulfate solution to 5.0mL of medium immediately prior to inoculation.

Use: For the cultivation of Desulfovibrio africanus and other Desulfovibrio species that prefer 2.5% NaCl.

Bacillus acidocaldarius Agar

Composition per liter:

Solution A

500.0mL

Solution B

500.0mL

pH 3.0–4.0 at 25°C

Solution A:

Composition per 500.0mL:

KH2PO4

3.0g

Yeast extract

1.0g

Glucose

1.0g

MgSO4·7H2O

0.5g

CaCl2·2H2O

0.25g

(NH4)2SO4

0.2g

Preparation of Solution A: Add components to distilled/deionized water and bring volume to 500.0mL. Adjust pH to 3.0–4.0. Mix thoroughly. Autoclave for 10 min at 15 psi pressure–121°C. Cool to 50°– 55°C.

Solution B:

Composition per 500.0mL:

Agar

30.0g

Preparation of Solution B: Add agar to distilled/deionized water and bring volume to 500.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C.

Preparation of Medium: Aseptically mix 500.0mL of solution A and 500.0mL of solution B. Mix thoroughly. Aseptically adjust pH to 3.0–4.0. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation and maintenance of Bacillus acidocaldarius.

Bacillus acidocaldarius Agar

Composition per liter:

Solution A

500.0mL

Solution B

500.0mL

pH 3.5 ± 0.5 at 25°C

Solution A:

Composition per 500.0mL:

Yeast extract

1.0g

KH2PO4

0.6g

MgSO4·7H2O

0.5g

CaCl2·2H2O

0.25g

(NH4)2·SO4

0.2g

Preparation of Solution A: Add components to distilled/deionized water and bring volume to 500.0mL. Mix thoroughly. Bring pH to 3.5. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C.

Solution B:

Composition per 500.0mL:

Agar

20.0g

Glucose

1.0g

Preparation of Solution B: Add components to distilled/deionized water and bring volume to 500.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C.

Preparation of Medium: Aseptically combine 500.0mL of solution A with 500.0mL of solution B. Mix thoroughly. Pour into sterile Petri dishes or aseptically distribute into sterile tubes.

Use: For the cultivation and maintenance of Alicyclobacillus acidocaldarius.

Bacillus acidoterrestris Agar

Composition per 1001.0mL:

Solution A

500.0mL

Solution C

500.0mL

Solution B (Trace elements solution SL-6)

1.0mL

pH 4.0 ± 0.2 at 25°C

Solution A:

Composition per 500.0mL:

Glucose

5.0g

KH2PO4

3.0g

Yeast extract

2.0g

MgSO4·7H2O

0.5g

CaCl2·2H2O

0.25g

(NH4)2SO4

0.2g

Preparation of Solution A: Add components to distilled/deionized water and bring volume to 500.0mL. Mix thoroughly. Adjust pH to 4.0. Autoclave for 15 min at 15 psi pressure–121°C.

Solution C:

Composition per 500.0mL:

Agar

15.0g

Preparation of Solution C: Add agar to distilled/deionized water and bring volume to 500.0mL. Gently heat and bring to boiling. Auto-clave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C.

Solution B (Trace Elements Solution SL-6):

Composition per liter:

MnCl2·4H2O

0.5g

H3BO3

0.3g

CoCl2·6H2O

0.2g

ZnSO4·7H2O

0.1g

Na2MoO4·2H2O

0.03g

NiCl2·6H2O

0.02g

CuCl2·2H2O

0.01g

Preparation of Solution B (Trace Elements Solution SL-6):

Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C.

Preparation of Medium: Aseptically combine 500.0mL of sterile solution A, 500.0mL of sterile solution C, and 1.0mL of sterile solution B. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation and maintenance of Bacillus acidoterrestris, Alicyclobacillus acidoterrestris, and Alicyclobacillus cycloheptanicus.

Bacillus acidoterrestris Broth

Composition per 1001.0mL:

Solution A

1.0L

Solution B (Trace elements solution SL-6)

1.0mL

pH 4.0 ± 0.2 at 25°C

Solution A:

Composition per liter:

Glucose

5.0g

KH2PO4

3.0g

Yeast extract

2.0g

MgSO4·7H2O

0.5g

CaCl2·2H2O

0.25g

(NH4)2SO4

0.2g

Preparation of Solution A: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 4.0. Autoclave for 15 min at 15 psi pressure–121°C.

Solution B (Trace Elements Solution SL-6):

Composition per liter:

MnCl2·4H2O

0.5g

H3BO3

0.3g

CoCl2·6H2O

0.2g

ZnSO4·7H2O

0.1g

Na2MoO4·2H2O

0.03g

NiCl2·6H2O

0.02g

CuCl2·2H2O

0.01g

Preparation of Solution B (Trace Elements Solution SL-6):

Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C.

Preparation of Medium: Aseptically combine 1.0L of sterile solution A and 1.0mL of sterile solution B. Mix thoroughly. Aseptically distribute into sterile tubes or flasks.

Use: For the cultivation and maintenance of Bacillus acidoterrestris, Alicyclobacillus acidoterrestris, and Alicyclobacillus cycloheptanicus.

Bacillus Agar

Composition per liter:

Agar

20.0g

(NH4)2SO4

1.3g

Glucose

1.0g

Yeast extract

1.0g

KH2PO4

0.37g

MgSO4·7H2O

0.25g

CaCl2·2H2O

0.07g

FeCl3

0.02g

pH 4.0 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 500.0mL. Mix thoroughly. Gently heat and bring to boiling. Adjust pH to 3.5. Prepare a separate agar solution by adding 20.0g/500.0mL of distilled/deionized water. Autoclave solutions separately for 15 min at 15 psi pressure–121°C. Cool to 50°– 55°C. Aseptically combine both solutions. This procedure avoids acid hydrolysis of the agar. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation of acidophilic Bacillus species such as Bacillus acidocaldarius.

Bacillus Agar, Modified

Composition per liter:

Agar

20.0g

Glucose

1.0g

Yeast extract

1.0g

KH2PO4

0.6g

MgSO4·7H2O

0.5g

CaCl2·2H2O

0.25g

(NH4)2SO4

0.2g

pH 3.0–4.0 at 25°C

Preparation of Medium: Add components, except agar and glucose, to distilled/deionized water and bring volume to 500.0mL. Mix thoroughly. Gently heat and bring to boiling. Adjust pH to 3.5. Prepare a separate agar and glucose solution by adding 20.0g of agar and 1.0g of glucose to 500.0mL of distilled/deionized water. Autoclave solutions separately for 15 min at 15 psi pressure–121°C. Cool to 50°– 55°C. Aseptically combine both solutions. This procedure avoids acid hydrolysis of the agar. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation of acidophilic Bacillus species such as Bacillus acidocaldarius.

Bacillus Agar, 1/4 Strength

Composition per liter:

Agar

18.0g

Yeast extract

2.5g

Pancreatic digest of casein

1.0g

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation and maintenance of Bacillus megaterium.

Bacillus benzoevorans Agar

Composition per liter:

Agar

15.0g

Yeast extract

6.0g

Peptone

5.0g

NaCl

5.0g

Na2HPO4·12H2O

3.6g

Sodium benzoate

2.0g

Beef extract

1.0g

KH2PO4

0.98g

NH4Cl

0.5g

MgSO4·7H2O

0.03g

Trace elements solution

0.2mL

pH 7.0–7.2 at 25°C

Trace Elements Solution:

Composition per 100.0mL:

FeSO4·7H2O

0.1g

MnCl2·4H2O

0.1g

ZnSO4·7H2O

0.1g

Preparation of Trace Elements Solution: 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. 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 Bacillus benzoevorans.

Bacillus benzoevorans Agar

Composition per liter:

Modified Palleroni and Doudoroff mineral base medium

50.0mL

Enriched Cytophaga agar

450.0mL

Sodium benzoate solution

100.0mL

pH 7.0 ± 0.2 at 25°C

Modified Palleroni and Doudoroff Mineral Base Medium:

Composition per 500.0mL:

Agar

15.0g

Na2HPO4·12H2O

6.0g

KH2PO4

2.4g

NH4·Cl

1.0g

MgSO4·7H2O

0.5g

CaCl2·6H2O

0.01g

FeCl3·6H2O

0.01g

Preparation of Modified Palleroni and Doudoroff Mineral Base Medium: Add components to distilled/deionized water and bring volume to 450.0mL. Mix thoroughly. Adjust pH to 7.2. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure– 121°C. Cool to 50°C.

Enriched Cytophaga Agar:

Composition per 500.0mL:

Agar

15.0g

Pancreatic digest of casein

0.5g

Beef extract

0.5g

Yeast extract

0.5g

Sodium acetate

0.2g

Preparation of Enriched Cytophaga Agar: Add components to distilled/deionized water and bring volume to 450.0mL. Mix thoroughly. Adjust pH to 6.8. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C.

Sodium Benzoate Solution:

Composition per 100.0mL:

Sodium benzoate

5.0g

Preparation of Sodium Benzoate Solution: Add sodium benzoate to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Aseptically combine 450.0mL of modified Palleroni and Doudoroff mineral base medium, 450.0mL of enriched Cytophaga agar, and 100.0mL sodium benzoate solution. Mix thoroughly. Pour into sterile Petri dishes or aseptically distribute into sterile tubes.

Use: For the cultivation of Bacillus benzoevorans.

Bacillus Broth

Composition per liter:

(NH4)2SO4

1.3g

Glucose

1.0g Yeast extract

1.0g

KH2PO4

0.37g

MgSO4·7H2O

0.25g

CaCl2·2H2O

0.07g

FeCl3

0.02g

pH 4.0 ± 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 4.0 with 10N H2SO4. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the cultivation of acidophilic Bacillus species such as Bacillus acidocaldarius.

Bacillus Broth, 1/4 Strength

Composition per liter:

Yeast extract

2.5g

Pancreatic digest of casein

1.0g

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the cultivation of Bacillus megaterium.

Bacillus cereus Agar Base with Egg Yolk Emulsion and Polymyxin

Composition per liter:

Agar

15.0g

Sodium pyruvate

10.0g

Mannitol

10.0g

Na2HPO4

2.5g

NaCl

2.0g

Peptone

1.0g

KH2PO4

0.25g

Bromthymol Blue

0.12g

MgSO4·7H2O

0.1g

Egg yolk emulsion

100.0mL

Selective supplement solution

10.0mL

pH 7.2 ± 0.2 at 25°C

Source: This medium, without egg yolk emulsion, is available as a premixed powder from HiMedia.

Selective Supplement Solution:

Composition per 10.0mL:

Polymyxin B

100,000 U

Preparation of Selective Supplement Solution: Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Egg Yolk Emulsion:

Composition per liter:

Egg yolks

30.0mL

NaCl, 0.9% solution

70.0mL

Preparation of Egg Yolk Emulsion: Soak eggs with 1:100 dilution of saturated mercuric chloride solution for 1 min. Crack 11 eggs and separate yolks from whites. Mix egg yolks. Measure 30.0mL of egg yolk emulsion and add to 70.0mL of 0.9% sterile NaCl solution. Mix thoroughly. Warm to 45°–50°C.

Preparation of Medium: Add components, except egg yolk emulsion, and selective supplement solution, to distilled/deionized water and bring volume to 890.0mL. 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 egg yolk emulsion and 10.0mL sterile selective supplement solution. Mix well. Pour into sterile Petri dishes or sterile tubes.

Use: For the isolation, detection, and enumeration of Bacillus cereus.

Bacillus cereus HiVeg Agar Base with Egg Yolk Emulsion

Composition per liter:

Agar

15.0g

Sodium pyruvate

10.0g

Mannitol

10.0g

Na2HPO4

2.5g

NaCl

2.0g

Plant peptone

1.0g

KH2PO4

0.25g

Bromthymol Blue

0.12g

MgSO4·7H2O

0.1g

Egg yolk emulsion

100.0mL

pH 7.2 ± 0.2 at 25°C

Source: This medium, without egg yolk emulsion, is available as a premixed powder from HiMedia.

Egg Yolk Emulsion:

Composition per liter:

Egg yolks

30.0mL

NaCl, 0.9% solution

70.0mL

Preparation of Egg Yolk Emulsion: Soak eggs with 1:100 dilution of saturated mercuric chloride solution for 1 min. Crack 11 eggs and separate yolks from whites. Mix egg yolks. Measure 30.0mL of egg yolk emulsion and add to 70.0mL of 0.9% sterile NaCl solution. Mix thoroughly. Warm to 45°–50°C.

Preparation of Medium: Add components, except egg yolk emulusion, to distilled/deionized water and bring volume 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 egg yolk emulsion. Mix well. Pour into sterile Petri dishes or sterile tubes.

Use: For the isolation, detection, and enumeration of Bacillus cereus.

Bacillus cereus Medium

(BCM)

Composition per 110.0mL:

Agar

2.0g

D-Mannitol

1.0g

(NH4)2PO4

0.1g

KCl

0.02g

MgSO4·7H2O

0.02g

Yeast extract

0.02g

Bromcresol Purple

4.0mg

Egg yolk emulsion, 20%

10.0mL

pH 7.0 ± 0.2 at 25°C

Egg Yolk Emulsion, 20%:

Composition per 100.0mL:

Chicken egg yolks

11

Whole chicken egg

1

NaCl (0.9% solution)

80.0mL

Preparation of Egg Yolk Emulsion, 20%: 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. Measure 20.0mL of egg yolk emulsion and add to 80.0mL of 0.9% NaCl solution. Mix thoroughly. Filter sterilize. Warm to 45°–50°C.

Preparation of Medium: Add components—except egg yolk emulsion, 20%—to distilled/deionized water and bring volume to 100.0mL. 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 of sterile egg yolk emulsion, 20%. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation of Bacillus cereus.

Bacillus cereus Motility Medium

See: BC Motility Medium

Bacillus cereus Selective Agar Base

Composition per liter:

Agar

15.0g

Sodium pyruvate

10.0g

Mannitol

10.0g

Na2HPO4

2.5g

NaCl

2.0g

Peptone

1.0g

KH2PO4

0.25g

Bromthymol Blue

0.12g

MgSO4·7H2O

0.1g

Egg yolk emulsion

25.0mL

Polymyxin B solution

10.0mL

pH 7.2 ± 0.2 at 25°C

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

Egg Yolk Emulsion:

Composition:

Chicken egg yolks

11

Whole chicken egg

1

Preparation of Egg Yolk Emulsion: 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.

Polymyxin B Solution:

Composition per 10.0mL:

Polymyxin B

100,000U

Preparation of Polymyxin B Solution: Add polymyxin B to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except egg yolk emulsion and polymyxin B solution, to distilled/deionized water and bring volume to 965.0mL. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Aseptically add sterile polymyxin B and 25.0mL of sterile egg yolk emulsion. Mix thoroughly. Pour into sterile Petri dishes or leave in tubes.

Use: For the selection and presumptive identification of Bacillus cereus. Also for the isolation and enumeration of these bacteria. Bacillus cereus grows as moderate-sized (5mm) crenated colonies, which are turquoise, surrounded by a precipitate of egg yolk, which is also turquoise.

Bacillus coagulans Medium

Composition per liter:

Agar

20.0g

Glucose

5.0g

Proteose peptone

5.0g

Yeast extract

5.0g

K2HPO4

4.0g

MnSO4·4H2O solution

10.0mL

CaCl2 solution

10.0mL

pH 5.0 ± 0.2 at 25°C

MnSO4·4H2O Solution:

Composition per 10.0mL:

MnSO4·4H2O

0.05mg

Preparation of MnSO4·4H2O Solution: Add MnSO4·4H2O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

CaCl2 Solution:

Composition per 10.0mL:

CaCl2

0.045mg

Preparation of CaCl2 Solution: Add CaCl2 to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except MnSO4·4H2O solution and CaCl2 solution, to distilled/deionized water and bring volume to 980.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Avoid overheating. Cool to 45°–50°C. Aseptically add sterile MnSO4·4H2O solution and CaCl2 solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation of Bacillus coagulans.

Bacillus cycloheptanicus Agar

Composition per 1001.0mL:

Solution A

500.0mL

Solution C

500.0mL

Solution B (Trace elements solution SL-6)

1.0mL

pH 4.0 ± 0.2 at 25°C

Solution A:

Composition per liter:

Yeast extract

5.0g

Glucose

5.0g

KH2PO4

3.0g

MgSO4·7H2O

0.5g

CaCl2·2H2O

0.25g

(NH4)2SO4

0.2g

Preparation of Solution A: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 4.0. Autoclave for 15 min at 15 psi pressure–121°C.

Solution C:

Composition per 500.0mL:

Agar

15.0g

Preparation of Solution C: Add agar to distilled/deionized water and bring volume to 500.0mL. Gently heat and bring to boiling. Auto-clave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C.

Solution B (Trace Elements Solution SL-6):

Composition per liter:

MnCl2·4H2O

0.5g

H3BO3

0.3g

CoCl2·6H2O

0.2g

ZnSO4·7H2O

0.1g

Na2MoO4·2H2O

0.03g

NiCl2·6H2O

0.02g

CuCl2·2H2O

0.01g

Preparation of Solution B (Trace Elements Solution SL-6):

Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C.

Preparation of Medium: Aseptically combine 500.0mL of sterile solution A, 500.0mL of sterile solution C, and 1.0mL of sterile solution

B. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation and maintenance of Bacillus cycloheptanicus, Alicyclobacillus acidoterrestris, and Alicyclobacillus cycloheptanicus.

Bacillus cycloheptanicus Broth

Composition per 1001.0mL:

Solution A

1.0L

Solution B (Trace elements solution SL-6)

1.0mL

pH 4.0 ± 0.2 at 25°C

Solution A:

Composition per liter:

Yeast extract

5.0g

Glucose

5.0g

KH2PO4

3.0g

MgSO4·7H2O

0.5g

CaCl2·2H2O

0.25g

(NH4)2SO4

0.2g

Preparation of Solution A: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 4.0. Autoclave for 15 min at 15 psi pressure–121°C.

Solution B (Trace Elements Solution SL-6):

Composition per liter:

MnCl2·4H2O

0.5g

H3BO3

0.3g

CoCl2·6H2O

0.2g

ZnSO4·7H2O

0.1g

Na2MoO4·2H2O

0.03g

NiCl2·6H2O

0.02g

CuCl2·2H2O

0.01g

Preparation of Solution B (Trace Elements Solution SL-6):

Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C.

Preparation of Medium: Aseptically combine 1.0L of sterile solution A and 1.0mL of sterile solution B. Mix thoroughly. Aseptically distribute into sterile tubes or flasks.

Use: For the cultivation of Bacillus cycloheptanicus, Alicyclobacillus acidoterrestris, and Alicyclobacillus cycloheptanicus.

Bacillus fastidiosus Agar

Composition per liter:

Allantoin

20.0g

Agar

15.0g

K2HPO4

0.8g

MgSO4·7H2O

0.5g

KH2PO4

0.2g

CaCl2·2H2O

50.0mg

FeSO4·7H2O

10.0mg

MnSO4·4H2O

1.0mg

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation of Bacillus fastidiosus.

Bacillus fastidiosus Medium

Composition per liter:

Agar

15.0g

Na2HPO4·12H2O

6.0g

Yeast extract

2.5g

Uric acid

1.0g

Mineral solution

100.0mL

pH 7.0 ± 0.2 at 25°C

Mineral Solution:

Composition per 100.0mL:

KH2PO4

0.1g

MgSO4·7H2O

0.03g

CaCl2

0.01g

NaCl

0.01g

FeCl3·6H2O

1.0mg

Preparation of Mineral Solution: 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. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation of Bacillus fastidiosus.

Bacillus filiformis Medium

(DSMZ Medium 992)

Composition per liter:

Yeast extract

10.0g

Sodium citrate

3.0g

KCl

2.0g

MgSO4·7H2O

1.0g

Sodium chloride solution

100.0mL

Sodium carbonate solution

10.0mL

Iron sulfate solution

1.0mL

Manganese chloride solution

1.0mL

pH 9.0 ± 0.2 at 25°C

Iron Sulfate Solution:

Composition per liter:

FeSO4·7H2O

50.0g

Preparation of Iron Sulfate Solution: Add FeSO4·7H2O to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

Sodium Chloride Solution:

Composition per 100.0mL:

NaCl

100.0g

Preparation of Sodium Chloride Solution: Add NaCl to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C.

Manganese Chloride Solution:

Composition per liter:

MnCl2·4H2O

0.36g

Preparation of Manganese Chloride Solution: Add MnCl2·4H2O to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

Sodium Carbonate Solution:

Composition per 10.0mL:

Na2CO3

3.0g

Preparation of Sodium Carbonate Solution: Add Na2CO3 to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C.

Preparation of Medium: Add components, except sodium chloride and sodium carbonate solutions, to distilled/deionized water and bring volume to 890.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Aseptically add 100.0mL sterile sodium chloride solution and 10.0mL sterile sodium carbonate solution. Mix thoroughly. Aseptically distribute into sterile tubes or flasks.

Use: For the cultivation of a Bacillus filiformis.

Bacillus halodenitrificans Agar

(LMG Medium 142)

Composition per liter:

NaCl

100.0g

Agar

15.0g

Sodium acetate·3H2O

10.0g

Na2HPO4

3.8g

KH2PO4

1.3g

(NH4)2SO4

1.0g

Mg(NO3)2·6H2O

1.0g

Yeast extract

1.0g

Magnesium nitrate solution

100.0mL

pH 7.2 ± 0.2 at 25°C

Magnesium Nitrate Solution:

Composition per 100.0mL:

Mg(NO3)2·6H2O

1.0g

Preparation of Magnesium Nitrate Solution: Add Mg(NO3)2·6H2O to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except magnesium nitrate solution, to distilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Adjust pH to 7.2 with KOH. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add 100.0mL sterile magnesium nitrate solution. Mix thoroughly. Aseptically pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation of Bacillus halodenitrificans.

Bacillus mascerans Medium

(TSBY Salt Medium)

(LMG 199)

Composition per liter:

NaCl

18.0g

Pancreatic digest of casein

17.0g

MgCl2·H2O

4.0g

MgSO4·7H2O

3.45g

Yeast extract

3.0g

Papaic digest of soybean meal

3.0g

K2HPO4

2.5g

Glucose

2.5g

KCl

0.34g

NH4Cl

0.25g

CaCl2·2H2O

0.14g

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

Use: For the cultivation of Bacillus mascerans, Carnobacterium alter-funditum, and Carnobacterium funditum.

Bacillus Medium

Composition per liter:

Agar

25.0g

Peptone

6.0g

Pancreatic digest of casein

3.0g

Yeast extract

3.0g

Beef extract

1.5g

MnSO4·4H2O

1.0μg

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 of Bacillus species.

Bacillus Medium

Composition per liter:

(NH4)2HPO4

1.0g

MgSO4·7H2O

0.2g

KCl

0.2g

Yeast extract

0.2g

Glucose solution

50.0mL

Bromcresol Purple solution

15.0mL

pH 7.0 ± 0.2 at 25°C

Glucose Solution:

Composition per 100.0mL:

Glucose

10.0g

Preparation of Glucose Solution: Add glucose to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

Bromcresol Purple Solution:

Composition per 20.0mL:

Bromcresol Purple

0.32g

Ethanol (95% solution)

20.0mL

Preparation of Bromcresol Purple Solution: Add Bromcresol Purple to 20.0mL of ethanol. Mix thoroughly.

Preparation of Medium: Add components, except glucose solution, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute 9.5mL volumes into test tubes that contain an inverted Durham tube. Autoclave for 20 min at 15 psi pressure–121°C. Cool to 25°C. Aseptically add 0.5mL of sterile glucose to each tube. Mix thoroughly.

Use: For cultivation and differentiation of Bacillus species based on acid and gas production from glucose.

Bacillus Medium

(ATCC Medium 21)

Composition per liter:

Glycerol

20.0g

L-Glutamic acid

4.0g

Citric acid

2.0g

K2HPO4

0.5g

Ferric ammonium citrate

0.5g

MgSO4

0.5g

pH 7.4 ± 0.2 at 25°C

Preparation of Medium: Add components to tap 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 Bacillus licheniformis.

Bacillus Medium

(ATCC Medium 455)

Composition per liter:

Soluble starch

30.0g

Agar

20.0g

Polypeptone™

5.0g

Yeast extract

5.0g

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Swirl medium to resuspend starch. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation and maintenance of Bacillus subtilis. Also used to detect amylase-producing microorganisms.

Bacillus Medium

(ATCC Medium 552)

Composition per liter:

Peptone

10.0g

Lactose

5.0g

NaCl

5.0g

Beef extract

3.0g

K2HPO4

2.0g

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the cultivation and maintenance of Bacillus species.

Bacillus pasteurii Agar

Composition per liter:

Agar

15.0g

Peptone

5.0g

NaCl

5.0g

Yeast extract

4.0g

Beef extract

1.0g

Urea solution

50.0mL

pH 8.0 ± 0.2 at 25°C

Urea Solution:

Composition per 100.0mL:

Urea

20.0g

Preparation of Urea Solution: Add urea to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Warm to 50°–55°C.

Preparation of Medium: Add components, except urea solution, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Aseptically add 50.0mL of sterile urea solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation and maintenance of Bacillus pasteurii.

Bacillus pasteurii Agar

Composition per liter:

Urea

20.0g

Agar

15.0g

Peptone

5.0g

Meat extract

3.0g

pH 7.0 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling. Adjust pH to 7.0. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation and maintenance of Bacillus pasteurii and Sporosarcina ureae.

Bacillus pasteurii Medium

Composition per liter:

Urea

20.0g

Agar

15.0g

Peptone

5.0g

NaCl

5.0g

Yeast extract

2.0g

Beef extract

1.0g

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 Bacillus pasteurii.

Bacillus pasteurii NH4 YE Medium

(Ammonium Yeast Extract Medium)

Composition per liter:

Yeast extract

20.0g

Agar

20.0g

(NH4)2SO4

10.0g

pH 9.0 ± 0.2 at 25°C

Preparation of Medium: Add each component to a separate flask and bring volume of each to 333.0mL with 0.13m Tris buffer, pH 9.0. Autoclave ingredients separately for 15 min at 15 psi pressure–121°C. No growth occurs if components are sterilized together. Cool to 50°– 55°C and aseptically combine solutions. Pour into sterile Petri dishes.

Use: For the cultivation and maintenance of Bacillus pasteurii.

Bacillus pasteurii Sporulation Agar

Composition per liter:

Urea

20.0g

Agar

15.0g

Peptone

5.0g

Meat extract

3.0g

MnSO4·H2O

10.0mg

pH 7.0 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling. Adjust pH to 7.0. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the induction of sporulation in various species, including Bacillus pasteurii and Sporosarcina ureae.

Bacillus polymyxa Agar

Composition per liter:

Agar

20.0g

Starch, soluble

10.0g

Peptone

5.0g

Yeast extract

5.0g

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation and maintenance of Bacillus macerans, Bacillus polymyxa, and Bacillus thermoglucosidasius.

Bacillus popilliae Maintenance Medium

Composition per liter:

Agar

20.0g

Yeast extract

15.0g

Pancreatic digest of casein

5.0g

K2HPO4

3.0g

Glucose solution

10.0mL

pH 7.2 ± 0.2 at 25°C

Glucose Solution:

Composition per 10.0mL:

Glucose

2.0g

Preparation of Glucose Solution: Add glucose to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except glucose solution, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add sterile glucose solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation and maintenance of Bacillus popilliae.

Bacillus popilliae Medium

Composition per liter:

Yeast extract

10.0g

Acid hydrolysate of casein

7.95g

K2HPO4

3.0g

Beef extract

1.36g

Trehalose

1.0g

Starch

0.68g

pH 7.3 ± 0.1 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat until dissolved. Do not overheat. Filter sterilize. Aseptically distribute into sterile tubes or flasks.

Use: For the cultivation of Bacillus popilliae.

Bacillus popilliae Medium

Composition per liter:

Yeast extract

15.0g

K2HPO4

3.0g

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the cultivation of Bacillus popilliae.

Bacillus Pullulan Salts

Composition per liter:

Pullulan

2.5g

NaCl

1.0g

NH4Cl

1.0g

KH2PO4

0.5g

MgSO4·7H2O

0.5g

Yeast extract

0.1g

CaCl2·2H2O

0.05g

Trace mineral solution

10.0mL

Vitamin solution

10.0mL

pH 6.0 ± 0.2 at 25°C

Trace Mineral Solution:

Composition per liter:

CoCl2·6H2O

0.2g

FeSO4·7H2O

0.13g

ZnCl2·2H2O

0.1g

MnCl2·4H2O

0.1g

CaCl2·2H2O

20.0mg

Na2SeO3

20.0mg

Na2WO4·2H2O

20.0mg

NaMoO4·2H2O

1.0mg

H3BO3

0.5mg

CuSO4·5H2O

0.4mg

KI

0.1mg

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

Vitamin Solution:

Composition per liter:

Pyridoxine·HCl

10.0mg

Thiamine·HCl

5.0mg

Riboflavin

5.0mg

Nicotinic acid

5.0mg Calcium pantothenate

5.0mg

p-Aminobenzoic acid

5.0mg

Thioctic acid

5.0mg

Biotin

2.0mg

Folic acid

2.0mg

Cyanocobalamin

0.1mg

Preparation of Vitamin Solution: Add components to distilled/ deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except vitamin solution, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Gently heat and bring to boiling. Adjust pH to 6.0. Auto-clave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Aseptically add sterile vitamin solution. Mix thoroughly. Aseptically distribute into sterile tubes or flasks.

Use: For the cultivation and maintenance of Bacillus species that can degrade pullulan.

Bacillus racemilacticus Agar

Composition per liter:

Agar

15.0g

CaCO3

5.0g

Glucose

5.0g

Peptone

5.0g

Yeast extract

5.0g

pH 6.8 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation and maintenance of Bacillus kaustophilus and Bacillus racemilacticus.

Bacillus racemilacticus Agar

Composition per liter:

Agar

15.0g

CaCO3

5.0g

Glucose

5.0g

Peptone

5.0g

Yeast extract

5.0g

pH 6.8 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling. Adjust pH to 6.8. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation and maintenance of Bacillus racemilacticus, Bacillus coagulans, Bacillus laevolacticus, and other Bacillus species.

Bacillus racemilacticus Agar

(YEPG with 0.5% CaCO3)

Composition per liter:

Agar

15.0g

CaCO3

5.0g

Glucose

5.0g

Peptone

5.0g

Yeast extract

5.0g

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation and maintenance of Bacillus racemilacticus and other Bacillus species.

Bacillus schlegelii Agar

(LMG Medium 85)

Composition per liter:

Agar

30.0g

Na2HPO4·12 H2O

9.0g

KH2PO4

1.5g

Sodium pyruvate

1.5g

NH4Cl

1.0g

MgSO4·7H2O

0.2g

MnSO4·H2O

10.0mg

CaCl2·2H2O

10.0mg

Ferric ammonium citrate

5.0mg

Trace elements solution

3.0mL

pH 7.1 ± 0.2 at 25°C

Trace Elements Solution:

Composition per liter:

H3BO3

0.3g

CoCl2·6H2O

0.2g

ZnSO4·7H2O

0.1g

Na2MoO4·2H2O

30.0mg

MnCl2·4H2O

30.0mg

NiCl2·6H2O

20.0mg

CuCl2·2H2O

10.0mg

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

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation of Bacillus schlegelii.

Bacillus schlegelii Agar

Composition per liter:

Noble agar

30.0g

Na2HPO4·2H2O

4.5g

KH2PO4

1.5g

NH4Cl

1.0g

MgSO4·7H2O

0.2g

MnSO4·H2O

0.01g

CaCl2·2H2O

0.01g

Ferric ammonium citrate

5.0mg

Agar solution

200.0mL

Pyruvate solution

100.0mL

Vrace elements solution SL-6

3.0mL

pH 7.1 ± 0.2 at 25°C

Agar Solution:

Composition per 200.0mL:

Noble agar

30.0g

Preparation of Agar Solution: Add agar to distilled/deionized water and bring volume to 200.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C.

Pyruvate Solution:

Composition per 100.0mL:

Sodium pyruvate

1.5g

Preparation of Pyruvate Solution: Add sodium pyruvate to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Warm to 45°–50°C.

Trace Elements Solution SL-6 :

Composition per liter:

H3BO3

0.3g

CoCl2·6H2O

0.2g

ZnSO4·7H2O

0.1g

MnCl2·4H2O

0.03g

Na2MoO4·H2O

0.03g

NiCl2·6H2O

0.02g

CuCl2·2H2O

0.01g

Preparation of Trace Elements Solution SL-6 : Add components to distilled/deionized water and bring volume to 1.0L. Adjust pH to 3.4.

Preparation of Medium: Add components, except sodium pyruvate solution and agar solution, to distilled/deionized water and bring volume to 700.0mL. Mix thoroughly. Gently heat and bring to boiling. Adjust pH to 7.1. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Add sodium pyruvate solution and agar solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation and maintenance of Bacillus schlegelii.

Bacillus schlegelii Broth

Composition per liter:

Na2HPO4·2H2O

4.5g

KH2PO4

1.5g

NH4Cl

1.0g

MgSO4·7H2O

0.2g

MnSO4·H2O

0.01g

CaCl2·2H2O

0.01g

Ferric ammonium citrate

5.0mg

Pyruvate solution

100.0mL

SL-6 trace elements

3.0mL

pH 7.1 ± 0.2 at 25°C

Pyruvate Solution:

Composition per 100.0mL:

Sodium pyruvate

1.5g

Preparation of Pyruvate Solution: Add sodium pyruvate to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

Trace Elements Solution SL-6 :

Composition per liter:

H3BO3

0.3g

CoCl2·6H2O

0.2g

ZnSO4·7H2O

0.1g

MnCl2·4H2O

0.03g

Na2MoO4·H2O

0.03g

NiCl2·6H2O

0.02g

CuCl2·2H2O

0.01g

Preparation of Trace Elements Solution SL-6 : Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 3.4.

Preparation of Medium: Add components, except sodium pyruvate solution, to distilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Gently heat and bring to boiling. Adjust pH to 7.1. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Aseptically add sodium pyruvate solution. Aseptically distribute into sterile tubes or flasks.

Use: For the cultivation and maintenance of Bacillus schlegelii.

Bacillus schlegelii Chemolithotrophic Growth Medium

(DSMZ Medium 261)

Composition per liter:

Na2HPO4·2H2O

4.5g

KH2PO4

1.5g

NH4Cl

1.0g

MgSO4·7H2O

0.2g

MnSO4·2H2O

0.01g

CaCl2·2H2O

0.01g

Ferric ammonium citrate

5.0mg

Trace elements solution SL-6

3.0mL

pH 7.1 ± 0.2 at 25°C

Trace Elements Solution SL-6:

Composition per liter:

MnCl2·4H2O

0.5g

H3BO3

0.3g

CoCl2·6H2O

0.2g

ZnSO4·7H2O

0.1g

Na2MoO4·2H2O

0.03g

NiCl2·6H2O

0.02g

CuCl2·2H2O

0.01g

Preparation of Trace Elements Solution SL-6: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.1. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure– 121°C.

Use: For the chemolithotrophic cultivation of Bacillus schlegelii. Incubation is at 65°C under an atmosphere of 5% O2, 10% CO2, 45% H2.

Bacillus schlegelii Heterotrophic Growth Medium

(DSMZ Medium 260)

Composition per liter:

Na2HPO4·2H2O

4.5g

Na-pyruvate

1.5g

KH2PO4

1.5g

NH4Cl

1.0g

MgSO4·7H2O

0.2g

MnSO4·2H2O

0.01g

CaCl2·2H2O

0.01g

Ferric ammonium citrate

5.0mg

Trace elements solution SL-6

3.0mL

pH 7.1 ± 0.2 at 25°C

Trace Elements Solution SL-6:

Composition per liter:

MnCl2·4H2O

0.5g

H3BO3

0.3g

CoCl2·6H2O

0.2g

ZnSO4·7H2O

0.1g

Na2MoO4·2H2O

0.03g

NiCl2·6H2O

0.02g

CuCl2·2H2O

0.01g

Preparation of Trace Elements Solution SL-6: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.1. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure– 121°C.

Use: For the heterotrophic cultivation of Bacillus schlegelii. Incubation is at 65°C.

Bacillus schlegelii Medium

(LMG Medium 85)

Composition per liter:

Na2HPO4·12 H2O

9.0g

KH2PO4

1.5g

Sodium pyruvate

1.5g

NH4Cl

1.0g

MgSO4·7H2O

0.2g

MnSO4·H2O

10.0mg

CaCl2·2H2O

10.0mg

Ferric ammonium citrate

5.0mg

Trace elements solution

3.0mL

pH 7.1 ± 0.2 at 25°C

Trace Elements Solution:

Composition per liter:

H3BO3

0.3g

CoCl2·6H2O

0.2g

ZnSO4·7H2O

0.1g

Na2MoO4·2H2O

30.0mg

MnCl2·4H2O

30.0mg

NiCl2·6H2O

20.0mg

CuCl2·2H2O

10.0mg

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

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute in 30–50 mL amounts in Erlenmeyer flasks. Autoclave for 15 min at 15 psi pressure–121°C. Incubate without agitation.

Use: For the cultivation of Bacillus schlegelii.

Bacillus selenitireducens Medium

(DSMZ Medium 968)

Composition per liter:

NaCl

90.0g

Na2CO3

10.6g

NaHCO3

4.2g

Na-lactate

1.70g

NaNO3

1.25g

Yeast extract

0.2g

K2HPO4

0.15g

(NH4)SO4

0.1g

KH2PO4

0.08g

MgSO4

25.0mg

Resazurin

0.5mg

Cysteine solution

10.0mL

Na2S·9H2O solution

10.0mL

Trace elements solution SL-10

1.0mL

Selenite tungstate solution

1.0mL

pH 9.8 ± 0.2 at 25°C

Cysteine Solution:

Composition per 10.0mL:

L-Cysteine·HCl·H2O

0.25g

Preparation of Cysteine Solution: Add L-cysteine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N2. Autoclave for 15 min at 15 psi pressure– 121°C.

Na2S·9H2O Solution:

Composition per 10.0mL:

Na2S·9H2O

0.25g

Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to distilled/deionized water and bring volume to 10.0mL. Sparge with N2. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.

Trace Elements Solution SL-10:

Composition per liter:

FeCl2·4H2O

1.5g

CoCl2·6H2O

190.0mg

MnCl2·4H2O

100.0mg

ZnCl2

70.0mg

Na2MoO4·2H2O

36.0mg

NiCl2·6H2O

24.0mg

H3BO3

6.0mg

CuCl2·2H2O

2.0mg

HCl (25% solution)

10.0mL

Preparation of Trace Elements Solution SL-10: Add FeCl2·4H2O to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized water and bring volume to 1.0L. Add remaining components. Mix thoroughly. Sparge with 80% N2 + 20% CO2. Autoclave for 15 min at 15 psi pressure–121°C.

Selenite Tungstate Solution

Composition per liter:

NaOH

0.5g

Na2WO4·2H2O

4.0mg

Na2SeO3·5H2O

3.0mg

Preparation of Selenite Tungstate Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N2. Filter sterilize.

Preparation of Medium: Add components, except NaHCO3, Na2CO3, cysteine solution, and Na2S·9H2O solution, to distilled/deionized water and bring volume to 980.0mL. Mix thoroughly. Gently heat and bring to boiling. Boil for 3 min. Cool to room temperature while sparging with 80% N2 gas. Add solid NaHCO3 and Na2CO3. Adjust pH to 9.8. Distribute to anaerobe tubes or bottles. Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature. Aseptically and anaerobically add per liter 10.0mL sterile cysteine solution and 10.0mL sterile Na2S·9H2O solution. Mix thoroughly.

Use: For the cultivation of Bacillus selenitireducens and Bacillus arseniciselenatis.

Bacillus stearothermophilus Broth

Composition per liter:

Pancreatic digest of casein

10.0g

Yeast extract

5.0g

K2HPO4

2.0g

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the cultivation of Bacillus stearothermophilus.

Bacillus stearothermophilus Defined Broth

Composition per 100.0mL:

Mineral salts solution

10.0mL

Potassium phosphate buffer

5.0mL

L-Glutamate·HCl (1% solution)

4.0mL

L-Leucine (1% solution)

1.64mL

L-Lysine·HCl (1% solution)

1.4mL

L-Serine (1% solution)

1.4mL

L-Aspartate (1% solution)

1.3mL

L-Valine (1% solution)

1.26mL

Biotin (0.01% solution)

1.0mL

Glucose (20% solution)

1.0mL

L-Isoleucine (1% solution)

1.0mL

L-Proline (1% solution)

1.0mL

Nicotinic acid (0.01% solution)

1.0mL

Thiamine·HCl (0.01% solution)

1.0mL

L-Phenylalanine (1% solution)

0.86mL

L-Alanine (1% solution)

0.84mL

L-Threonine (1% solution)

0.84mL

L-Arginine·HCl (1% solution)

0.64mL

L-Tyrosine (1% solution)

0.56mL

L-Methionine (1% solution)

0.52mL

Glycine (1% solution)

0.5mL

L-Asparagine·H2O (1% solution)

0.5mL

L-Cystine (1% solution)

0.5mL

L-Glutamine (1% solution)

0.5mL

L-Histidine·HCl·H2O (1% solution)

0.42mL

L-Tryptophan (1% solution)

0.3mL

CaCl2 (5% solution)

0.01mL

FeCl3·6H2O (0.05% solution)

0.01mL

MnCl2 (10mm solution)

0.01mL

ZnSO4·7H2O (5% solution)

0.01mL

pH 7.3 ± 0.2 at 25°C

Mineral Salts Solution:

Composition per liter:

NaCl

10.0g

NH4Cl

10.0g

MgSO4

4.0g

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

Potassium Phosphate Buffer:

Composition per 500.0mL:

K2HPO4

125.0g

KH2PO4

30.0g

Preparation of Potassium Phosphate Buffer: Add components to distilled/deionized water and bring volume to 500.0mL. Mix thoroughly.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

Use: For the cultivation of Bacillus stearothermophilus in a chemically defined medium.

Bacillus stearothermophilus Sporulation Broth

Composition per liter:

Agar

20.0g

Pancreatic digest of gelatin

5.0g

Yeast extract

4.0g

Beef extract

3.0g

MnCl2·4H2O

10.0μg

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation and sporulation of Bacillus stearothermophilus.

Bacillus thermoalcalophilus Medium

Composition per liter:

Yeast extract

10.0g

Sodium acetate

3.0g

KCl

1.8g

Na2SO4

0.4g

K2HPO4

0.3g

KH2PO4

0.3g

MgSO4

0.2g

FeSO4

0.01g

pH 8.2 ± 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 8.2. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure– 121°C.

Use: For the cultivation of Bacillus thermoalcalophilus.

Bacillus thermoantarcticus Medium

Composition per liter:

Yeast extract

6.0g

NaCl

3.0g

Soil extract

500.0mL

pH 5.6–5.8 at 25°C

Soil Extract:

Composition per liter:

Garden soil, air dried

400.0g

Preparation of Soil Extract: Pass 400.0g of air-dried garden soil through a coarse sieve. Add soil to 960.0mL of tap water. Mix thoroughly. Autoclave for 60 min at 15 psi pressure–121°C. Cool to room temperature. Allow residue to settle. Decant supernatant solution. Filter through Whatman filter paper. Distribute into bottles in 200.0mL volumes. Autoclave for 15 min at 15 psi pressure–121°C. Store at room temperature until clear.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 5.6–5.8. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure– 121°C.

Use: For the cultivation of Bacillus thermoantarcticus.

Bacillus thermoglucosidasius Agar

Composition per liter:

Agar

30.0g

Starch

10.0g

Peptone

5.0g

Beef extract

3.0g

K2HPO4

3.0g

Yeast extract

3.0g

pH 7.0 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.0. Gently heat and bring to boiling. Distribute into tubes or flasks. Auto-clave for 20 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation and maintenance of Bacillus thermoglucosidasius.

Bacillus thermoglucosidasius Agar

Composition per liter:

Agar

30.0g

Soluble starch

10.0g

Peptone

5.0g

Beef extract

3.0g

KH2PO4

3.0g

Yeast extract

3.0g

pH 7.0 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling. Adjust pH to 7.0. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation and maintenance of Bacillus thermoglucosidasius.

Bacillus thermoleovorans Medium

Composition per liter:

n-Heptadecane

1.0g

(NH4)2HPO4

1.0g

Yeast extract

1.0g

KCl

0.2g

MgSO4·7H2O

0.2g

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 Bacillus thermoleovorans.

Bacillus thuringiensis Medium

Composition per liter:

Glucose

3.0g

(NH4)2SO4

2.0g

Yeast extract

2.0g

K2HPO4·3H2O

0.5g

MgSO4·7H2O

0.2g

CaCl2·2H2O

0.08g

MnSO4·4H2O

0.05g

pH 7.3 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.3. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure– 121°C.

Use: For the cultivation of Bacillus thuringiensis.

Bacillus tusciae Medium

Composition per liter:

Na2HPO4·2H2O

2.9g

KH2PO4

2.3g

NH4Cl

1.0g

MgSO4·7H2O

0.5g

NaHCO3

0.5g

Fe(NH4) citrate

0.05g

CaCl2·2H2O

0.01g

MnSO4·H2O

0.01g

Ferric ammonium citrate solution

20.0mL

Trace elements solution SL-6

5.0mL

pH 4.0 ± 0.2 at 25°C

Ferric Ammonium Citrate Solution:

Composition per 20.0mL:

Ferric ammonium citrate

0.05g

Preparation of Ferric Ammonium Citrate Solution: Add ferric ammonium citrate to distilled/deionized water and bring volume to 20.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure– 121°C.

Trace Elements Solution SL-6:

Composition per liter:

MnCl2·4H2O

0.5g

H3BO3

0.3g

CoCl2·6H2O

0.2g

ZnSO4·7H2O

0.1g

Na2MoO4·2H2O

0.03g

NiCl2·6H2O

0.02g

CuCl2·2H2O

0.01g

Preparation of Trace Elements Solution SL-6: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C.

Preparation of Medium: Add components, except ferric ammonium citrate solution and trace elements solution SL-6, to distilled/deionized water and bring volume to 975.0mL. Mix thoroughly. Gently heat and bring to boiling. Adjust pH to 4.0. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Aseptically add 20.0mL of sterile ferric ammonium citrate solution and 5.0mL of sterile trace elements solution SL-6. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. For chemolithotropic growth, incubate the culture under 2% O2 + 10% CO2 + 60% H2 + 28% N2.

Use: For the chemolithotrophic growth of Bacillus tusciae.

Bacillus tusciae Medium

Composition per liter:

Agar

15.0g

Na2HPO4·2H2O

2.9g

KH2PO4

2.3g

NH4Cl

1.0g

MgSO4·7H2O

0.5g

NaHCO3

0.5g

Fe(NH4) citrate

0.05g

CaCl2·2H2O

0.01g

MnSO4·H2O

0.01g

Ferric ammonium citrate solution

20.0mL

Carbon source

10.0mL

Trace elements solution SL-6

5.0mL

pH 4.0 ± 0.2 at 25°C

Ferric Ammonium Citrate Solution:

Composition per 20.0mL:

Ferric ammonium citrate

0.05g

Preparation of Ferric Ammonium Citrate Solution: Add ferric ammonium citrate to distilled/deionized water and bring volume to 20.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure– 121°C.

Carbon Source:

Composition per 10.0mL:

Carbohydrate

2.0g

Organic acid (alternate)

1.0g

Preparation of Carbon Source: Add either carbohydrate or organic acid to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Trace Elements Solution SL-6:

Composition per liter:

MnCl2·4H2O

0.5g

H3BO3

0.3g

CoCl2·6H2O

0.2g

ZnSO4·7H2O

0.1g

Na2MoO4·2H2O

0.03g

NiCl2·6H2O

0.02g

CuCl2·2H2O

0.01g

Preparation of Trace Elements Solution SL-6: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C.

Preparation of Medium: Add components, except ferric ammonium citrate solution, trace elements solution SL-6, and carbon source, to distilled/deionized water and bring volume to 965.0mL. Mix thoroughly. Gently heat and bring to boiling. Adjust pH to 4.0. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Aseptically add 20.0mL of sterile ferric ammonium citrate solution, 10.0mL of sterile carbon source, and 5.0mL of sterile trace elements solution SL-6. Mix thoroughly. Aseptically distribute into sterile tubes or flasks.

Use: For the heterotrophic growth of Bacillus tusciae.

Bacillus Xylose Salts

Composition per liter:

Yeast extract

5.0g

Xylose

5.0g

NaCl

1.0g

NH4Cl

1.0g

KH2PO4

0.5g

MgSO4·7H2O

0.5g

CaCl2·2H2O

0.05g

Trace mineral solution

10.0mL

Vitamin solution

10.0mL

pH 4.0 ± 0.2 at 25°C

Trace Mineral Solution:

Composition per liter:

CoCl2·6H2O

0.2g

FeSO4·7H2O

0.13g

ZnCl2·2H2O

0.1g

MnCl2·4H2O

0.1g

CaCl2·2H2O

20.0mg

Na2SeO3

20.0mg

Na2WO4·2H2O

20.0mg

NaMoO4·2H2O

1.0mg

H3BO3

0.5mg

CuSO4·5H2O

0.4mg

KI

0.1mg

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

Vitamin Solution:

Composition per liter:

Pyridoxine·HCl

10.0mg

Thiamine·HCl

5.0mg

Riboflavin

5.0mg

Nicotinic acid

5.0mg

Calcium pantothenate

5.0mg

p-Aminobenzoic acid

5.0mg

Thioctic acid

5.0mg

Biotin

2.0mg

Folic acid

2.0mg

Cyanocobalamin

0.1mg

Preparation of Vitamin Solution: Add components to distilled/ deionized water and bring volume to 1.0L. 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. Adjust pH of medium to 4.0. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the cultivation and maintenance of Bacillus species that can utilize xylose as a carbon source.

Bacterial Cell Agar

(BCA)

Composition per liter:

Tryptose

17.36g

Agar

15.0g

NaCl

8.68g

Beef extract

5,2g

Yeast extract

1.7g

pH 7.3 ± 0.2 at 25°C

Preparation of Medium: Add components, except agar, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 30°C. Inoculate with a culture of Aeromonas hydrophila. Incubate with shaking at 30°C for 72 hr. Centrifuge culture in 40.0mL volumes at 10,000 × g for 10 min. Wash the cells four times in sterile 0.85% saline. Resuspend the cell pellet in 25.0mL of distilled/deionized water. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. In a separate flask, add 15.0g of agar to 1.0L of distilled/deionized water. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically combine 25.0mL of washed cells and 250.0mL of cooled, sterile agar solution. Mix thoroughly. Pour into sterile Petri dishes.

Use: For the cultivation of freshwater Myxobacterium species.

Bacterium Medium

Composition per liter:

Agar

20.0g

Peptone

6.0g

Yeast extract

3.0g

Beef extract

1.5g

Glucose

1.0g

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.

Use: An archaic medium used for the cultivation and growth of bacteria originally classified in the genus Bacterium but now classified in the genera Brevibacterium and Kurthia.

Bacteroides Bile Esculin Agar

(BBE Agar)

Composition per liter:

Oxgall

20.0g

Pancreatic digest of casein

15.0g

Agar

15.0g

Papaic digest of soybean meal

5.0g

NaCl

5.0g

Esculin

1.0g

Ferric ammonium citrate

0.5g

Gentamicin solution

2.5mL

Hemin solution

2.5mL

Vitamin K1 solution

1.0mL

pH 7.0 ± 0.2 at 25°C

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

Gentamicin Solution:

Composition per 10.0mL:

Gentamicin

0.4mg

Preparation of Gentamicin Solution: Add gentamicin to 10.0mL of distilled/deionized water. Mix thoroughly. Filter sterilize.

Hemin Solution:

Composition per 100.0mL:

Hemin

0.5g

NaOH (1N solution)

10.0mL

Preparation of Hemin Solution: Add components to 100.0mL of distilled/deionized water. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C.

Vitamin K1 Solution:

Composition per 100.0mL:

Vitamin K1

1.0g

Ethanol

99.0mL

Preparation of Vitamin K1 Solution: Add vitamin K1 to 99.0mL of absolute ethanol. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except hemin solution, gentamicin solution, and vitamin K1 solution, to distilled/deionized water and bring volume to 994.0mL. 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 2.5mL of sterile hemin solution, 2.5mL of sterile gentamicin solution, and 1.0mL of sterile vitamin K1 solution.

Use: For the selection and presumptive identification of the Bacteri-odes fragilis group. For the differentiation of Bacteroides species based on the hydrolysis of esculin and presence of catalase. After incubation for 48 hr, bacteria of the Bacteroides fragilis group appear as gray, circular, raised colonies larger than 1.0mm. Esculin hydrolysis is indicated by the presence of a blackened zone around the colonies.

Bacteroides cellulosolvens Medium

Composition per liter:

Cellobiose or cellulose

5.0g

NaHCO3

2.0g

NH4Cl

0.68g

K2HPO4

0.3g

L-Cysteine·HCl·H2O

0.25g

Na2S·9H2O

0.25g

KH2PO4

0.18g

(NH4)2SO4

0.15g

MgSO4·7H2O

0.12g

CaCl2·2H2O

0.06g

FeSO4·7H2O

0.02g

Resazurin

1.0mg

Trace elements solution

10.0mL

Vitamin solution

10.0mL

pH 7.0 ± 0.2 at 25°C

Trace Elements Solution:

Composition per liter:

MgSO4·7H2O

3.0g

Nitrilotriacetic acid

1.5 g

CaCl2·2H2O

.1.0g

NaCl

1.0g

MnSO4·2H2O

0.5 g

CoSO4·7H2O

0.18 g

ZnSO4·7H2O

0.18 g

FeSO4·7H2O

0.1g

NiCl2·6H2O

0.025 g

KAI(SO4)2·12H2O

0.02g

CuSO4·5H2O

0.01g

H3BO3

0.01g

Na2MoO4·2H2O

0.01g

Na2SeO3·5H2O

0.3 mg

Preparation of Trace Elements Solution: Add nitrilotriacetic acid to approximately 500.0mL distilled/deionized water. Dissolve by adding KOH and adjust pH to 6.5. Add remaining components. Bring volume to 1.0L with additional distilled/deionized water. Adjust pH to 7.0 with KOH.

Vitamin Solution:

Composition per liter:

Pyridoxine·HCl

10.0mg

Calcium DL-pantothenate

5.0mg

Lipoic acid

5.0mg

Nicotinic acid

5.0mg

p-Aminobenzoic acid

5.0mg

Riboflavin

5.0mg

Thiamine·HCl

5.0mg

Biotin

2.0mg

Folic acid

2.0mg

Vitamin B12

0.1mg

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

Cellobiose Solution:

Composition per 50.0mL:

D-Cellobiose (or cellulose)

5.0g

Preparation of Cellobiose Solution: Add cellobiose (or cellulose) to distilled/deionized water and bring volume to 50.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except cellobiose solution, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically add 50.0mL of sterile cellobiose (or cellulose) solution. Mix thoroughly. Aseptically distribute into sterile tubes or flasks.

Use: For the cultivation and maintenance of Bacteroides cellosolvens.

Bacteroides HiVeg Agar Base

with Selective Supplement

(BBE)

Composition per liter:

Plant hydrolysate

25.0g

Agar

15.0g

Papaic digest of soybean meal

10.0g

NaCl

5.0g

Synthetic detergent No. II

2.0g

Esculin

1.0g

Ferric ammonium citrate

0.5g

Fe4(P2O7)3·H2O

0.01g

Vitamin K1

0.01g

Selective supplement solution

10.0mL

pH 7.2 ± 0.2 at 25°C

Source: This medium, without selective supplement, is available as a premixed powder from HiMedia.

Selective Supplement Solution:

Composition per 10.0mL:

Gentamicin

0.1mg

Preparation of Selective Supplement Solution: Add gentamicin to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except selective supplement, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Heat with frequent agitation and boil for 1 min to completely dissolve. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Add 10.0mL of sterile selective supplement. Mix thoroughly. Pour into sterile Petri dishes or leave in tubes.

Use: For the selection and presumptive identification of the Bacteri-odes fragilis group. For the differentiation of Bacteroides species based on the hydrolysis of esculin and presence of catalase. After incubation for 48 hr, bacteria of the Bacteroides fragilis group appear as gray, circular, raised colonies larger than 1.0mm. Esculin hydrolysis is indicated by the presence of a blackened zone around the colonies.

Bacteroides Medium

Composition per liter:

Pancreatic digest of casein

27.0g

Yeast extract

3.0g

K2HPO4

2.5g

K2CO3

2.0g

NaCl

2.0g

Hemin solution

10.0mL

Vitamin K1 solution

0.2mL

Hemin Solution:

Composition per 100.0mL:

Hemin

1.0g

NaOH (1N solution)

20.0mL

Preparation of Hemin Solution: Add hemin to 20.0mL of 1N NaOH solution. Mix thoroughly. Bring volume to 100.0mL with distilled/deionized water.

Vitamin K1 Solution:

Composition per 100.0mL:

Vitamin K1

1.0g

Ethanol

99.0mL

Preparation of Vitamin K1 Solution: Add vitamin K1 to 99.0mL of absolute ethanol. Mix thoroughly.

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 Bacteroides asaccharolyticus and Bacteroides melaninogenicus.

Bacteroides nodosus Agar

Composition per liter:

Agar

14.0g

Liver hydrolysate

10.0g

Proteose peptone No. 3

10.0g

Trypsin 1:250

10.0g

NaCl

5.0g

Yeast extract

2.0g

L-Cysteine·HCl solution

2.5mL

pH 7.4 ± 0.2 at 25°C

L-Cysteine·HCl Solution:

Composition per 10.0mL:

L-Cysteine HCl

1.0g

Preparation of L-Cysteine·HCl Solution: Dissolve 1.0g of L-cysteine·HCl in distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Warm to 50°C.

Preparation of Medium: Add components, except agar and L-cysteine·HCl solution, to distilled/deionized water and bring volume to 997.5mL. Mix thoroughly. Adjust pH to 8.5. Gently heat and bring to boiling. Boil for 5 min. Filter and allow to cool to 25°C. Adjust pH to 7.4. Add 14.0g of agar. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Aseptically add 2.5mL of sterile L-cysteine·HCl solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation and maintenance of Bacteroides nodosus.

Bacteroides vulgatus Medium

(LMG Medium 204)

Composition per liter:

Special peptone

23.0g

Agar

15.0g

Glucose

5.0g

NaCl

5.0g

Soluble starch

1.0g

Cysteine hydrochloride

0.3g

Horse blood, sterile defibrinated

50.0mL

pH 7.1 ± 0.2 at 25°C

Preparation of Medium: Add components, except horse blood, to 950.0mL 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 45°–50°C. Aseptically add 50.0mL sterile horse blood. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation and maintenance of Bacteroides vulgatus.

BAF Agar

Composition per liter:

Glucose

30.0g

Agar

15.0g

Peptone

2.0g

KH2PO4

0.5g

MgSO4·7H2O

0.5g

Yeast extract

0.2g

CaCl2·2H2O

100.0mg

FeCl3·6H2O

10.0mg

MnSO4

5.0mg

ZnSO4·7H2O

1.0mg

Folic acid

100.0μg

Inositol

50.0μg

Thiamine·HCl

50.0μg

Biotin

1.0μg

pH 5.8 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling. Adjust pH to 5.8. 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 a wide variety of bacteria.

BAGG Broth

(Buffered Azide Glucose Glycerol Broth)

Composition per liter:

Pancreatic digest of casein

10.0g

Peptic digest of animal tissue

10.0g

Glucose

5.0g

NaCl

5.0g

K2HPO4

4.0g

KH2PO4

1.5g

NaN3

0.5g

Bromcresol Purple

0.015g

Glycerol

5.0mL

pH 6.9 ± 0.2 at 25°C

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

Caution: Sodium azide is toxic. Azides also react with metals and disposal must be highly diluted.

Preparation of Medium: Add 5.0mL of glycerol to 900.0mL of distilled/deionized water. Add remaining components and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes in 10.0mL volumes. Autoclave for 15 min at 10 psi pressure–116°C.

Use: For the cultivation of fecal streptococci from a variety of clinical and nonclinical specimens. It is recommended for qualitative presumptive and confirmatory tests for fecal streptococci.

BAGG Broth Base with Glycerol

(Buffered Azide Glucose Glycerol Broth Base)

Composition per liter:

Tryptose

20.0g

Glucose

5.0g

NaCl

5.0g

K2HPO4

4.0g

KH2PO4

1.5g

NaN3

0.5g

Bromcresol Purple

0.015g

Glycerol

5.0mL

pH 6.9 ± 0.2 at 25°C

Source: This medium without glycerol is available as a premixed powder from HiMedia.

Caution: Sodium azide is toxic. Azides also react with metals and disposal must be highly diluted.

Preparation of Medium: Add 5.0mL of glycerol to 900.0mL of distilled/deionized water. Add remaining components and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes in 10.0mL volumes. Autoclave for 15 min at 10 psi pressure–115°C.

Use: For the cultivation of fecal streptococci from a variety of clinical and nonclinical specimens. It is recommended for qualitative presumptive and confirmatory tests for fecal streptococci.

BAGG HiVeg Broth Base with Glycerol

(Buffered Azide Glucose Glycerol HiVeg Broth Base)

Composition per liter:

Plant hydrolysate No. 1

20.0g

Glucose

5.0g

NaCl

5.0g

K2HPO4

4.0g

KH2PO4

1.5g

NaN3

0.5g

Bromcresol Purple

0.015g

Glycerol

5.0mL

pH 6.9 ± 0.2 at 25°C

Source: This medium without glycerol is available as a premixed powder from HiMedia.

Caution: Sodium azide is toxic. Azides also react with metals and disposal must be highly diluted.

Preparation of Medium: Add 5.0mL of glycerol to 900.0mL of distilled/deionized water. Add remaining components and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes in 10.0mL volumes. Autoclave for 15 min at 10 psi pressure–115°C.

Use: For the cultivation of fecal streptococci from a variety of clinical and nonclinical specimens. It is recommended for qualitative presumptive and confirmatory tests for fecal streptococci.

Baird-Parker Agar

Composition per liter:

Agar

17.0g

Glycine

12.0g

Sodium pyruvate

10.0g

Pancreatic digest of casein

10.0g

Beef extract

5.0g

LiCl

5.0g

Yeast extract

1.0g

pH 7.0 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Oxoid Unipath and BD Diagnostic Systems.

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 45°–50°C. Pour into sterile Petri dishes.

Use: Used as a base for the preparation of egg-tellurite-glycine-pyruvate agar for the selective isolation and enumeration of coagulase-positive staphylococci from food, skin, soil, air, and other materials.

Baird-Parker Agar

Composition per liter:

Agar

17.0g

Glycine

12.0g

Sodium pyruvate

10.0g

Pancreatic digest of casein

10.0g

Beef extract

5.0g

LiCl

5.0g

Yeast extract

1.0g

Sulfamethazine solution

10.0mL

pH 7.0 ± 0.2 at 25°C

Sulfamethazine Solution:

Composition per 10.0mL:

Sulfamethazine

0.05g

Preparation of Sulfamethazine Solution: Add sulfamethazine to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except sulfamethazine solution, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add sterile sulfamethazine solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: Used as a base for the preparation of egg-tellurite-glycine-pyruvate agar for the selective isolation and enumeration of coagulase-positive staphylococci from food, skin, soil, air, and other materials.

Baird-Parker Agar Base with Egg Yolk Tellurite Enrichment

Composition per liter:

Agar

20.0g

Glycine

12.0g

Casein enzymatic hydrolysate

10.0g

Sodium pyruvate

10.0g

Plant extract

5.0g

LiCl

5.0g

Yeast extract

1.0g

Egg yolk tellurite enrichment

50.0mL

pH 7.0 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Hi-Media.

Caution: Lithium chloride is harmful. Avoid bodily contact and inhalation of vapors. On contact with skin wash with plenty of water immediately.

Egg Yolk Tellurite Enrichment:

Composition per 100.0mL:

Chicken egg yolks

10

K2TeO3

0.15g

NaCl (0.9% solution)

50.0mL

Preparation of Egg Yolk Tellurite Enrichment: Soak eggs with 1:100 dilution of saturated mercuric chloride solution for 1 min. Crack 11 eggs and separate yolks from whites. Mix egg yolks. Measure 30.0mL of egg yolk emulsion and add to 70.0mL of 0.9% NaCl solution. Mix thoroughly. Add 0.15g K2TeO3. Filter sterilize. Warm to 45°– 50°C.

Caution: Potassium tellurite is toxic.

Preparation of Medium: Add components, except egg yolk tellu-rite enrichment, to distilled/deionized water and bring volume to 950.0mL. 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 50 mL of egg yolk tellurite enrichment. Mix well. Pour into sterile Petri dishes or sterile tubes.

Use: For the selective isolation and enumeration of coagulase-positive staphylococci.

Baird-Parker Agar Base, HiVeg

with Egg Yolk Tellurite Enrichment

Composition per liter:

Agar

20.0g

Glycine

12.0g

Plant hydrolysate

10.0g

Sodium pyruvate

10.0g

Plant extract

5.0g

LiCl

5.0g

Yeast extract

1.0g

Egg yolk tellurite enrichment

50.0mL

pH 7.0 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Hi-Media.

Caution: Lithium chloride is harmful. Avoid bodily contact and inhalation of vapors. On contact with skin wash with plenty of water immediately.

Egg Yolk Tellurite Enrichment:

Composition per 100.0mL:

Chicken egg yolks

10

K2TeO3

0.15g

NaCl (0.9% solution)

50.0mL

Preparation of Egg Yolk Tellurite Enrichment: Soak eggs with 1:100 dilution of saturated mercuric chloride solution for 1 min. Crack 11 eggs and separate yolks from whites. Mix egg yolks. Measure 30.0mL of egg yolk emulsion and add to 70.0mL of 0.9% NaCl solution. Mix thoroughly. Add 0.15g K2TeO3. Filter sterilize. Warm to 45°– 50°C.

Caution: Potassium tellurite is toxic.

Preparation of Medium: Add components, except egg yolk tellu-rite enrichment, to distilled/deionized water and bring volume to 950.0mL. 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 50 mL of egg yolk tellurite enrichment. Mix well. Pour into sterile Petri dishes or sterile tubes.

Use: For the selective isolation and enumeration of coagulase-positive staphylococci.

Baird-Parker Agar, Supplemented

Composition per liter:

Agar

17.0g

Glycine

12.0g

Sodium pyruvate

10.0g

Pancreatic digest of casein

10.0g

Beef extract

5.0g

LiCl

5.0g

Yeast extract

1.0g

RPF supplement

100.0mL

pH 7.0 ± 0.2 at 25°C

RPF Supplement:

Composition per 100.0mL:

Bovine fibrinogen

3.75g

Trypsin inhibitor

25.0mg

K2TeO3

25.0mg

Rabbit plasma

25.0mL

Caution: Potassium tellurite is toxic.

Preparation of RPF Supplement: Add components to distilled/ deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except RPF supplement, to distilled/deionized water and bring volume 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 filter-sterilized RPF supplement. Mix thoroughly but gently. Pour into sterile Petri dishes.

Use: For the selective isolation and enumeration of coagulase-positive staphylococci from food, skin, soil, air, and other materials. For the differentiation and identification of staphylococci on the basis of their ability to coagulate plasma. Colonies surrounded by an opaque zone of coagulated plasma are diagnostic for Staphylococcus aureus.

Baird-Parker Egg Yolk Agar

(ISO)

Composition per 1050.0mL:

Agar

20.0g

L-Glycine

12.0g

Pancreatic digest of casein

10.0g

Sodium pyruvate

10.0g

Meat extract

5.0g

LiCl

5.0g

Yeast extract

1.0g

Egg yolk tellurite enrichment

50.0mL

pH 7.2 ± 0.2 at 25°C

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

Caution: Lithium chloride is harmful. Avoid bodily contact and inhalation of vapors. On contact with skin wash with plenty of water immediately.

Egg Yolk Tellurite Enrichment:

Composition per 100.0mL:

Chicken egg yolks

10

K2TeO3

0.15g

NaCl (0.9% solution)

50.0mL

Preparation of Egg Yolk Tellurite Enrichment: Soak eggs with 1:100 dilution of saturated mercuric chloride solution for 1 min. Crack 11 eggs and separate yolks from whites. Mix egg yolks. Measure 30.0mL of egg yolk emulsion and add to 70.0mL of 0.9% NaCl solu-tion. Mix thoroughly. Add 0.15g K2TeO3. Filter sterilize. Warm to 45°– 50°C.

Caution: Potassium tellurite is toxic.

Preparation of Medium: Add components, except egg yolk tellu-rite enrichment, to distilled/deionized water and bring volume to 950.0mL. 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 50 mL of egg yolk tellurite enrichment. Mix well. Pour into sterile Petri dishes or sterile tubes.

Use: For the selective isolation and enumeration of coagulase-positive staphylococci. A selective medium for the isolation and enumeration of coagulase-positive staphylococci from food, with formulation conforming to that recommended in ISO 6888-1:1999.

Baird-Parker Medium

(BAM M17)

Composition per liter:

Agar

20.0g

Glycine

12.0g

Sodium pyruvate

10.0g

Pancreatic digest of casein

10.0g

Beef extract

5.0g

LiCl·6H2O

5.0g

Yeast extract

1.0g

Egg yolk tellurite enrichment

50.0mL

pH 7.0 ± 0.2 at 25°C

Egg Yolk Tellurite Enrichment:

Composition per 100.0mL:

Chicken egg yolks

10

K2TeO3

0.15g

NaCl (0.9% solution)

50.0mL

Preparation of Egg Yolk Tellurite Enrichment: Soak eggs with 1:100 dilution of saturated mercuric chloride solution for 1 min. Crack 11 eggs and separate yolks from whites. Mix egg yolks. Measure 30.0mL of egg yolk emulsion and add to 70.0mL of 0.9% NaCl solution. Mix thoroughly. Add 0.15g K2TeO3. Filter sterilize. Warm to 45°– 50°C.

Caution: Potassium tellurite is toxic.

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

Preparation of Medium: Add components, except EY tellurite enrichment, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 48°–50°C. Aseptically add 50.0mL of sterile EY tellurite enrichment. Mix thoroughly. Pour into sterile Petri dishes. The medium must be densely opaque. Dry plates before use. Plates can be stored for up to 5 days at 20–25°C before use.

Use: For the selective isolation and enumeration of coagulase-positive staphylococci from foods.

Baird-Parker Medium

(BAM M17)

Composition per liter:

Agar

20.0g

Glycine

12.0g

Sodium pyruvate

10.0g

Pancreatic digest of casein

10.0g Beef extract

5.0g

LiCl·6H2O

5.0g

Yeast extract

1.0g

Egg yolk tellurite enrichment

50.0mL

pH 7.0 ± 0.2 at 25°C

Egg Yolk Tellurite Enrichment:

Composition per 100.0mL:

Chicken egg yolks

10

K2TeO3

0.15g

NaCl (0.9% solution)

50.0mL

Preparation of Egg Yolk Tellurite Enrichment: Soak eggs with 1:100 dilution of saturated mercuric chloride solution for 1 min. Crack 11 eggs and separate yolks from whites. Mix egg yolks. Measure 30.0mL of egg yolk emulsion and add to 70.0mL of 0.9% NaCl solution. Mix thoroughly. Add 0.15g K2TeO3. Filter sterilize. Warm to 45°– 50°C.

Caution: Potassium tellurite is toxic.

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

Preparation of Medium: Add components, except egg yolk tellu-rite enrichment, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 48°–50°C. Aseptically add 50.0mL of sterile egg yolk tellurite enrichment. Mix thoroughly. Pour into sterile Petri dishes. The medium must be densely opqaue. Dry plates before use. Plates can be stored for up to 5 days at 20–25°C before use.

Use: For the selective isolation and enumeration of coagulase-positive staphylococci from foods.

Balamuth Medium

Composition per 200.0mL:

Dehydrated egg yolk

36.0g

Dried liver concentrate

1.0g

Rice starch

0.2g

Potassium phosphate buffer, pH 7.5

125.0mL

NaCl solution

125.0mL

pH 7.3 ± 0.2 at 25°C

NaCl Solution

Composition per 200.0mL:

NaCl

1.6g

Preparation of NaCl Solution: Add NaCl to distilled/deionized water and bring volume to 200.0mL. Mix thoroughly.

Potassium Phosphate Buffer, 0.067 M

Composition per 200.0mL:

K2HPO4 (1M solution)

8.6mL

KH2PO4 (1M solution)

4.66mL

Preparation of Potassium Phosphate Buffer: Combine the K2HPO4 and KH2PO4 solutions. Bring volume to 200.0mL with distilled/deionized water. Adjust pH to 7.5.

Preparation of Medium: Add dehydrated egg yolk to 36.0mL of distilled/deionized water. Add 125.0mL of 0.8% NaCl. Mix thoroughly in a blender. Heat in a covered, double boiler until infusion reaches 80°C and maintain at this temperature for 20 min. Add 20.0mL of distilled/deionized H2O. Filter through a layer of cheesecloth. To 90– 100.0mL of filtrate add 0.8% NaCl solution to bring volume to 125.0mL. Autoclave for 20 min at 15 psi pressure–121°C. Cool to 4°C. Filter. To filtrate, add an equal volume of 0.067M potassium phosphate buffer, pH 7.5. Add 1.0g of dried liver concentrate. Mix thoroughly. Distribute into tubes or flasks in 10.0mL volumes. Autoclave for 20 min at 15 psi pressure–121°C. Prior to inoculation, add 0.01g of rice starch to each tube.

Use: For the cultivation and maintenance of Entamoeba histolytica and other intestinal protozoa.

BAM Agar

(ATCC Medium 1655)

Composition per liter:

Agar

30.0g

Glucose

5.0g

KH2PO4

3.0g

Yeast extract

1.0g

MgSO4·7H2O

0.5g

CaCl2·2H2O

0.25g

(NH4)2SO4

0.2g

Trace elements

1.0mL

pH 4.0 ± 0.2 at 25°C

Trace Elements:

Composition per liter:

CaCl2·2H2O

0.66g

Na2MoO4·2H2O

0.3g

ZnSO4·7H2O

0.18g

CoCl2·6H2O

0.18g

CuSO4·5H2O

0.16g

MnSO4·4H2O

0.15g

H3BO3

0.1g

Preparation of Trace Elements: Add components to 1.0L of distilled/deionized water. Mix thoroughly.

Preparation of Medium: Add components, except agar, to distilled/deionized water and bring volume to 800.0mL. Mix thoroughly. Gently heat and bring to boiling. Adjust medium to pH 4.0 with H2SO4. Add agar to 200.0mL of distilled/deionized water. Autoclave agar separately to avoid acid hydrolysis. Autoclave for 15 min at 15 psi pressure–121°C. Mix the two solutions together. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation and maintenance of Bacillus acidoterrestris.

BAM Broth

Composition per liter:

Glucose

5.0g

KH2PO4

3.0g

Yeast extract

1.0g

MgSO4·7H2O

0.5g

CaCl2·2H2O

0.25g

(NH4)2SO4

0.2g

Trace elements

1.0mL

pH 4.0 ± 0.2 at 25°C

Trace Elements:

Composition per liter:

CaCl2·2H2O

0.66g

Na2MoO4·2H2O

0.3g

ZnSO4·7H2O

0.18g

CoCl2·6H2O

0.18g

CuSO4·5H2O

0.16g MnSO4·4H2O

0.15g

H3BO3

0.1g

Preparation of Trace Elements: Add components to 1.0L of distilled/deionized water. 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. Adjust medium to pH 4.0 with H2SO4. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the cultivation and maintenance of Bacillus acidoterrestris.

BAM SM Agar

(ATCC Medium 1656)

Composition per liter:

Agar

20.0g

Yeast extract

6.0g

Glucose

5.0g

KH2PO4

3.0g

MgSO4·7H2O

0.5g

CaCl2·2H2O

0.25g

(NH4)2SO4

0.2g

Trace elements

1.0mL

pH 4.0 ± 0.2 at 25°C

Trace Elements:

Composition per liter:

CaCl2·2H2O

0.66g

Na2MoO4·2H2O

0.3g

ZnSO4·7H2O

0.18g

CoCl2·6H2O

0.18g

CuSO4·5H2O

0.16g

MnSO4·4H2O

0.15g

H3BO3

0.1g

Preparation of Trace Elements: Add components to 1.0L of distilled/deionized water. Mix thoroughly.

Preparation of Medium: Add components, except agar, to distilled/deionized water and bring volume to 800.0mL. Mix thoroughly. Gently heat and bring to boiling. Adjust medium to pH 4.0 with H2SO4. Add agar to 200.0mL of distilled/deionized water. Autoclave agar separately to avoid acid hydrolysis. Autoclave for 15 min at 15 psi pressure–121°C. Mix the two solutions together. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation and maintenance of Bacillus cycloheptanicus.

BAM SM Agar, Modified

Composition per liter:

Agar

30.0g

Glucose

5.0g

KH2PO4

3.0g

Yeast extract

1.0g

MgSO4·7H2O

0.5g

CaCl2·2H2O

0.25g

(NH4)2SO4

0.2g

Trace elements

1.0mL

pH 4.0 ± 0.2 at 25°C

Trace Elements:

Composition per liter:

CaCl2·2H2O

0.66g

Na2MoO4·2H2O

0.30g

ZnSO4·7H2O

0.18g

CoCl2·6H2O

0.18g

CuSO4·5H2O

0.16g

MnSO4·4H2O

0.15g

H3BO3

0.10g

Preparation of Trace Elements: Add components to 1.0L of distilled/deionized water. Mix thoroughly.

Preparation of Medium: Add components, except agar, to distilled/deionized water and bring volume to 800.0mL. Mix thoroughly. Gently heat and bring to boiling. Adjust medium to pH 4.0 with H2SO4. Add agar to 200.0mL of distilled/deionized water. Autoclave agar separately to avoid acid hydrolysis. Autoclave for 15 min at 15 psi pressure–121°C. Mix the two solutions together. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation and maintenance of Bacillus cycloheptanicus.

BAM SM Broth

Composition per liter:

Yeast extract

6.0g

Glucose

5.0g

KH2PO4

3.0g

MgSO4·7H2O

0.5g

CaCl2·2H2O

0.25g

(NH4)2SO4

0.2g

Trace elements

1.0mL

pH 4.0 ± 0.2 at 25°C

Trace Elements:

Composition per liter:

CaCl2·2H2O

0.66g

Na2MoO4·2H2O

0.3g

ZnSO4·7H2O

0.18g

CoCl2·6H2O

0.18g

CuSO4·5H2O

0.16g

MnSO4·4H2O

0.15g

H3BO3

0.1g

Preparation of Trace Elements: Add components to 1.0L of distilled/deionized water. 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. Adjust medium to pH 4.0 with H2SO4. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the cultivation and maintenance of Bacillus cycloheptanicus.

Bandoni’s MYP Medium

Composition per liter:

Agar

15.0g

Malt extract

7.0g

Papaic digest of soybean meal

1.0g

Yeast extract

0.5g

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 or leave in tubes.

Use: For the cultivation and maintenance of Coleosporium tussilaginis and Cystofilobasidium capitatum.

Basal Medium

(DSMZ Medium 1001)

Composition per liter:

NaHCO3

2.5g

NH4Cl

0.25g

NaH2PO4

0.6g

KCl

1.0g

Iron nitrilotriacetic acid solution

20.0mL

Vitamin mix

10.0mL

Mineral mix

10.0mL

Sodium acetate solution

10.0mL

pH 6.9 ± 0.2 at 25°C

Mineral Mix:

Composition per liter:

MgSO4·7H2O

3.0g

Nitrilotriacetic acid

1.5g

NaCl

1.0g

MnSO4·2H2O

0.5g

ZnCl2

0.13g

CoCl2·6H2O

0.1g

CaCl2·2H2O

0.1g

FeSO4·7H2O

0.1g

Na2MoO4·4H2O

0.025g

NaWO4·2H2O

0.025g

NiCl2·6H2O

0.024g

CuSO4·5H2O

0.01g

KAl(SO4)2·12H2O

0.01g

H3BO3

0.01g

Preparation of Mineral Mix: Add nitrilotriacetic acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH to 6.5 with KOH. Add remaining components. Add distilled/deionized water to 1.0L. Mix thoroughly.

Vitamin Mix:

Composition per liter:

Pyridoxine-HCl

10.0mg

Thiamine-HCl·2H2O

5.0mg

Riboflavin

5.0mg

Nicotinic acid

5.0mg

D-Ca-pantothenate

5.0mg

p-Aminobenzoic acid

5.0mg

Thioctic acid

5.0mg

Biotin

2.0mg

Folic acid

2.0mg

Vitamin B12

0.1mg

Preparation of Vitamin Mix: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 80% H2 + 20% CO2. Filter sterilize.

Sodium Acetate Solution:

Composition per 100.0mL:

Sodium acetate

13.6g

Preparation of Sodium Acetate Solution: Add sodium acetate to distilled/deionized water and bring volume to 80.0mL with distilled/ deionized water. Mix thoroughly. Bring volume to 100.0mL with distilled/deionized water. Sparge with 100% N2 for 45 min. Seal in bottle. Autoclave for 15 min at 15 psi pressure–121°C.

Iron Nitriloacetic Acid Solution:

Composition per 100.0mL:

FeCl3·6H2O

13.5g

Sodium nitrilotriacetic acid (NTA)

12.8g

NaHCO3

8.2g

Preparation of Iron Nitriloacetic Acid: Add NaHCO3 to distilled/deionized water and bring volume to 70.0mL with distilled/deionized water. Mix thoroughly. Add NTA. Mix thoroughly. Add FeCl3·6H2O. Adjust pH to 6.5 using 10N NaOH. Bring volume to 100.0mL with distilled/deionized water. Stir for about 15 minutes to allow components to go into solution. Sparge with 100% N2 for 45 min. Filter sterilize. Aseptically and anoxically dispense into sterile serum bottles.

Preparation of Medium: Add components, except iron nitriloacetic acid solution and sodium acetate solution, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Bubble the medium with 80:20 N2:CO2 (final pH should be 6.8 to 7.0). Approximately 10.0mL of media (anaerobic culture tube) should be gassed for 5 min in the aqueous phase (bubbled) and the headspace gassed for 1 min e prior to sealing the container. Autoclave for 15 min at 15 psi pressure– 121°C. Add electron donor (acetate-final conc. of 10mM-) and electron acceptor (Fe(III)NTA (final conc. of 10mM), from sterile, anaerobic stock solutions using a sterile syringe and needle flushed with anaerobic gas. This medium should not be exposed to direct sunlight!

Use: For the cultivation of a Rhodoferax ferrireducens.

Basal Mineral Medium

Composition per liter:

NH4Cl

0.8g

K2HPO4

0.7g

MgSO4·7H2O

0.01g

Disodium EDTA

9.2mg

FeSO4·7H2O

7.0mg

CaSO4·2H2O

2.0mg

H3BO3

0.1mg

ZnSO4·7H2O

0.1mg

MnSO4·4H2O

0.02mg

Co(NO3)2

0.01mg

NaMoO4·2H2O

0.01mg

CuSO4·5H2O

0.5μg

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize.

Use: For the cultivation of Beggiatoa species.

Basal Synthetic Medium

Composition per liter:

L-Glutamic acid

20.0g

(NH4)2SO4

4.0g

K2HPO4

1.88g

KH2PO4

0.57g

MgSO4·7H2O

0.2g

Salt solution

10.0mL

Salt Solution:

Composition per liter:

FeCl3·6H2O

0.6g

MnCl2·4H2O

0.6g

ZnCl2

0.6g CuSO4·5H2O

0.6g

CaCl2·2H2O

0.6g

NaCl

0.6g

Preparation of Salt Solution: Add components to 1.0L of distilled/deionized water. 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. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the cultivation and maintenance of Acinetobacter lwoffii.

Basal Thermophile Medium

Composition per liter:

Solution 1

850.0mL

Solution 2

100.0mL

Solution 3

50.0mL

Solution 1:

Composition per 850.0mL:

Pancreatic digest of casein

10.0g

K2HPO4

1.5g

NH4Cl

0.9g

KH2PO4

0.75g

MgCl2·6H2O

0.2g

Trace elements solution

9.0mL

Wolfe’s vitamin solution

5.0mL

Resazurin (0.2% solution)

1.0mL

FeSO4·7H2O (10% solution)

0.03mL

Preparation of Solution 1: Add components to distilled/deionized water and bring volume to 850.0mL. Mix thoroughly. Autoclave for 45 min at 15 psi pressure–121°C. Cool to 45°–50°C.

Solution 2:

Composition per 100.0mL:

Yeast extract

3.0g

Preparation of Solution 2: Add yeast extract to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 45 min at 15 psi pressure–121°C. Cool to 45°–50°C.

Solution 3:

Composition per 50.0mL:

Glucose

5.0g

Preparation of Solution 3: Add glucose to distilled/deionized water and bring volume to 50.0mL. Mix thoroughly. Autoclave for 45 min at 15 psi pressure–121°C. Cool to 45°–50°C.

Trace Elements Solution:

Composition per liter:

Nitrilotriacetic acid

12.5g

NaCl

1.0g

FeCl3·4H2O

0.2g

MnCl2·4H2O

0.1g

CaCl2·2H2O

0.1g

ZnCl2

0.1g

CuCl2

0.02g

Na2SeO3

0.02g

CoCl2·6H2O

0.017g

H3BO3

0.01g

Na2MoO4·2H2O

0.01g

Preparation of Trace Elements Solution: Add nitrilotriacetic acid to 100.0mL of distilled/deionized water. Adjust pH to 6.5 with KOH. Add remaining components and bring volume to 1.0L. Mix thoroughly.

Wolfe’s Vitamin Solution:

Composition per liter:

Pyridoxine·HCl

10.0mg

Thiamine·HCl

5.0mg

Riboflavin

5.0mg

Nicotinic acid

5.0mg

Calcium pantothenate

5.0mg

p-Aminobenzoic acid

5.0mg

Thioctic acid

5.0mg

Biotin

2.0mg

Folic acid

2.0mg

Cyanocobalamin

0.1mg

Preparation of Wolfe’s Vitamin Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

Na2S·9H2O Solution:

Composition per 100.0mL:

Na2S·9H2O

10.0g

Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C.

Preparation of Medium: Aseptically combine solution 1, solution 2, and solution 3 under 100% N2. Distribute into tubes in 10.0mL volumes under 100% N2. Immediately prior to inoculation, aseptically add 0.1mL of sterile Na2S·9H2O solution to each tube.

Use: For the cultivation and maintenance of Clostridium species, Fervidobacterium nodosum, and Thermoanaerobium brockii.

Base Agar

See: Antibiotic Medium 2

Base Agar with Low pH

See: Antibiotic Medium 8

Base Cholesterol Medium

Composition per liter:

Casitone

10.0g

Yeast extract

10.0g

Cholesterol, ash free

2.0g

CaCl2

1.0g

Lecithin, type IV

1.0g

Sodium thioglycolate

0.5g

Resazurin

1.0mg

Preparation of Medium: Prepare and dispense medium under 100% N2. Add cholesterol and lecithin to distilled/deionized water and bring volume to 200.0mL of water. Mix thoroughly. Sparge with 100% N2 for 10 min. Add other components to distilled/deionized water and bring volume to 800.0mL of water. Mix thoroughly. Combine the two solutions. Adjust pH to 7.5 with KOH. Gently heat and bring to boiling. Continue boiling while sparging with 100% N2 until the resazurin turns colorless. Cool under 100% N2. Distribute into tubes or flasks. Auto-clave for 15 min at 15 psi pressure–121°C. Mix well after autoclaving.

Use: For the cultivation of Eubacterium coprostanoligenes.

Base Layer Agar with Nutrient Overlay Agar

Composition per 2.5L: Fat substrate

50.0g

Nutrient agar

1.5L

Basal medium

1.0L

Fat Substrate:

Composition:

Fat

50.0g

Preparation of Fat Substrate: Tributyrin, corn oil, soybean oil, any cooking oil, lard, tallow, or triglycerides that do not contain anti-oxidants or other inhibitory substances may be used. Remove free fatty acids in the fat substrate by dissolving 50.0g of fat substrate in 500.0mL of petroleum ether. Pass the solution through an activated alumina column. Remove the petroleum ether by evaporation on a steam table under 100% N2. Autoclave for 30 min at 15 psi pressure–121°C. Cool to 50°C.

Nutrient Agar:

Composition per liter:

Agar

15.0g

Pancreatic digest of gelatin

5.0g

Beef extract

3.0g

Preparation of Nutrient Agar: 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. Cool to 45°–50°C.

Source: The medium is available as a premixed powder from BD Diagnostic Systems.

Basal Medium:

Composition per liter:

Agar

15.0g

Victoria Blue B solution

200.0mL

Preparation of Basal Medium: Add agar to 800.0mL of distilled/ deionized water. If tributyrin is used as the fat substrate, add agar to 1.0L of distilled/deionized water. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. If tributyrin is not used as the fat substrate, aseptically add 200.0mL of Victoria Blue B solution. Mix thoroughly.

Victoria Blue B Solution:

Composition per 200.0mL:

Victoria Blue B

0.12g

Preparation of Victoria Blue B Solution: Add the Victoria Blue B to 200.0mL of distilled/deionized water. Mix thoroughly. Filter sterilize. Warm to 50°C.

Preparation of Medium: Aseptically combine 1.0L of sterile basal medium with 50.0g of sterile fat substrate in a warm, sterile blender container. Blend for 1 min until homogenized. Rapidly pour into sterile Petri dishes in 7.0mL volumes. Dry the surface of the plates by partially opening the lids in a laminar flow hood for 15 min. Add dilution of food samples to be tested. When the inoculum is dry, pour nutrient agar as an overlay onto each plate. Use 10–12mL of nutrient agar per plate.

Use: For the isolation, cultivation, and identification of lipolytic microorganisms from food.

Basic Cultivation Medium

Composition per liter:

Yeast extract

10.0g

Glucose

5.0g

(NH4)2PO4

1.5g

K2HPO4

1.0g

MgSO4·7H2O

0.2g

Fe2(SO4)3·5H2O

0.01g

ZnSO4·7H2O

0.002g

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

Use: For the cultivation of a wide variety of microorganisms.

Basic Mineral Medium

Composition per liter:

NH4NO3

2.5g

Na2HPO4·2H2O

1.0g

MgSO4·7H2O

0.5g

Fe(SO4)3·5H2O

0.01g

Co(NO3)2·6H2O

0.005g

CaCl2·2H2O

1.0mg

KH2PO4

0.5mg

MnSO4·2H2O

0.1mg

(NH4)6Mo7O24·4H2O

0.1mg

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: To supply the mineral nutrients necessary for the cultivation of a wide variety of microorganisms. Various carbon sources can be added as sterilized solutions for testing carbon utilization capabilities.

BBE Agar

See: Bacteroides Bile Esculin Agar

BBGS Agar

See: Bile Salts Brilliant Green Starch Agar

BC Medium

(Medium for Acetivibrio cellulolyticus)

Composition per liter:

Cellulose powder

3.0g

NaHCO3

2.0g

Mineral solution 1

75.0mL

Mineral solution 2

75.0mL

Cysteine-sulfide reducing solution

12.8mL

FeSO4·7H2O solution

10.0mL

Vitamin mixture

10.0mL

Wolfe’s mineral solution

10.0mL

Resazurin (0.1% solution)

1.0mL

pH 7.6 ± 0.2 at 25°C

Caution: This medium contains sodium sulfide and may produce toxic H2S gas. Prepare in a chemical fume hood.

Mineral Solution 1:

Composition per liter:

K2HPO4

3.9g

Preparation of Mineral Solution 1: Add K2HPO4 to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

Mineral Solution 2:

Composition per liter:

NH4Cl

12.0g

Na2SO4

2.5g KH2PO4

2.4g

MgSO4·7H2O

1.2g

CaCl2·2H2O

0.8g

Preparation of Mineral Solution 2: Add components to distilled/ deionized water and bring volume to 1.0L. Mix thoroughly.

FeSO4·7H2O Solution:

Composition per 100.0mL:

FeSO4·7H2O

0.2g

Preparation of FeSO4·7H2O Solution: Dissolve FeSO4·7H2O in 100.0mL of distilled/deionized water. Add three drops of concentrated HCl. Mix thoroughly.

Vitamin Mixture:

Composition per liter:

Pyridoxine·HCl

10.0mg

Thiamine·HCl

5.0mg

Cyanocobalamin

5.0mg

Lipoic acid (thioctic acid)

5.0mg

Biotin

2.0mg

p-Aminobenzoic acid

0.5mg

Preparation of Vitamin Mixture: Add components to distilled/ deionized water and bring volume to 1.0L. Store below −20°C.

Wolfe’s Mineral Solution:

Composition per liter MgSO4·7H2O

3.0g

Nitriloacetic acid

1.5g

MnSO4·H2O

0.5g

NaCl

1.0g

FeSO4 ·7H2O

0.1g

CoCl2·6H2O

0.1g

CaCl2

0.1g

ZnSO4·7H2O

0.1g

CuSO4·5H2O

0.01g

AlK(SO4)2·12H2O

0.01g

H3BO3

0.01g

Na2MoO4·2H2O

0.01g

Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic acid to 500.0mL of distilled/deionized water and adjust to pH 6.5 with KOH to dissolve. Bring volume to 1.0L with distilled/deionized water. Add remaining components one at a time.

Cysteine-Sulfide Reducing Solution:

Composition per 200.0mL:

L-Cysteine·HCl·H2O

2.5g

Na2S·9H2O

2.5g

Preparation of Cysteine-Sulfide Reducing Solution: Add Lcysteine·HCl·H2O to 50.0mL of distilled/deionized water. Quickly adjust pH to 10 with fresh 3N NaOH and flush under 100% N2. Add Na2S·9H2O. Bring volume to 200.0mL with distilled/deionized water. Boil under 100% N2. Transfer anaerobically to tubes or flasks and stopper. Autoclave for 15 min at 15 psi pressure–121°C.

Preparation of Medium: Add cellulose and NaHCO3 to distilled/deionized water and bring volume to 800.0mL. Add all other components except cysteine-sulfide reducing solution. Heat and boil under 90% N2 + 10% CO2. Cool and continue flushing under 90% N2 + 10% CO2. The pH should be 7.6 at room temperature; do not adjust. Add 8.0mL of cysteinesulfide reducing solution. Add 4.8mL more of cysteine-sulfide reducing solution. Distribute anaerobically into tubes in 7.0mL volumes and cap.

Use: For the cultivation and maintenance of Acetivibrio cellulolyticus, Acetivibrio cellulosolvens, Bacteroides cellulosolvens, and other cellulose-degrading microorganisms.

BC Motility Medium

( Bacillus cereus Motility Medium)

Composition per liter:

Pancreatic digest of casein

10.0g

Glucose

5.0g

Agar

3.0g

Na2HPO4

2.5g

Yeast extract

2.5g

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 in 2.0mL volumes. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the cultivation and observation of motility of Bacillus cereus.

BC Motility Test HiVeg Medium

Composition per liter:

Plant hydrolysate

10.0g

Glucose

5.0g

Agar

3.0g

Na2HPO4

2.5g

Yeast extract

2.5g

pH 7.4 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Hi-Media.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes in 2.0mL volumes. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the cultivation and observation of motility of Bacillus cereus.

BCA

See: Bacterial Cell Agar

BCG Glucose Agar

(Snyder Test Agar)

Composition per liter:

Agar

20.0g

Glucose

20.0g

Peptic digest of animal tissue

20.0g

NaCl

5.0g

Bromcresol Green

0.02

pH 4.8 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Hi-Media.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes in 10.0mL volumes. Autoclave for 15 min at 15 psi pressure–121°C. Do not overheat. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation and enumeration of lactobacilli in saliva and indication of dental caries activity.

BCG Glucose HiVeg Agar

(Snyder Test HiVeg Agar)

Composition per liter:

Agar

20.0g

Glucose

20.0g

Plant peptone

20.0g

NaCl

5.0g

Bromcresol Green

0.02

pH 4.8 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Hi-Media.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes in 10.0mL volumes. Autoclave for 15 min at 15 psi pressure–121°C. Do not overheat. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation and enumeration of lactobacilli in saliva and indication of dental caries activity.

BCM

See: Bacillus cereus Medium

BCM Bacillus cereus Group Plating Medium

Composition per liter: Proprietary

Source: This medium is available from Biosynth International, Inc.

Use: For detection of Bacillus cereus in food. The medium contains 5-bromo-4-chloro-3-indoxyl-myoinositol-1-phosphate as a chromogenic substrate, which changes from colorless to turquoise upon enzymatic cleavage. B. cereus, B. mycoides, B. thuringiensis, and B. weihenstephanensis secrete phosphatidylinositol phospholipase C and so grow as turquoise colonies with species-specific morphologies.

BCM O157:H7(+) Plating Medium

Composition per liter: Proprietary

Source: This medium is available from Biosynth International, Inc.

Use: For detection of this highly pathogenic EHEC serovar BCM O157:H7(+).

BCP Azide Broth

(Bromcresol Purple Azide Broth)

Composition per liter:

Casein peptone

10.0g

Yeast extract

10.0g

D-Glucose

5.0g

NaCl

5.0g

K2HPO4

2.7g

KH2PO4

2.7g

NaN3

0.5g

Bromcresol Purple

0.032g

pH 6.9 ± 0.2 at 25°C

Caution: Sodium azide is toxic. Azides also react with metals and disposal must be highly diluted.

Preparation of Medium: Add components to distilled/deionized water to 1.0L. Mix thoroughly. Gently heat to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For use in the confirmation test for the presence of fecal streptococci in water and wastewater.

BCP Broth

See : Bromcresol Purple Dextrose Broth

BCP D Agar

(Bromcresol Purple Deoxycholate Agar)

Composition per liter:

Agar

25.0g

Lactose

10.0g

Sucrose

10.0g

Pancreatic digest of casein

7.5g

Thiopeptone

7.5g

NaCl

5.0g

Yeast extract

2.0g

Sodium citrate

2.0g

Sodium deoxycholate

1.0g

Bromcresol Purple

0.02g

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. Pour into sterile Petri dishes without sterilization. Do not autoclave. Use the same day.

Use: For the isolation, cultivation, and differentiation of Gram-negative enteric bacilli from clinical and nonclinical specimens. For the isolation, cultivation, and identification of microorganisms from fecal specimens. For the isolation and cultivation of Salmonella, Shigella, and other nonlactose- and nonsucrose-fermenting microorganisms. Nonlactose/nonsucrose fermenting microorganisms appear as colorless or blue colonies. Lactose/ sucrose-fermenting microorganisms, such as coliform bacteria, appear as yellow-opaque white colonies surrounded by a zone of precipitated deoxycholate.

BCP DCLS Agar

(Bromcresol Purple Deoxycholate

Citrate Lactose Sucrose Agar)

Composition per liter:

Agar

14.0g

Sodium citrate

10.0g

Lactose

7.5g

Sucrose

7.5g

Pancreatic digest of casein

7.5g

Peptone

7.5g

NaCl

5.0g

Na2S2O3·5H2O

5.0g

Yeast extract

3.0g

Meat extract

3.0g

Sodium deoxycholate

2.5g

Bromcresol Purple

0.02g

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. Pour into sterile Petri dishes without sterilization. Do not autoclave. Use the same day.

Use: For the differential isolation of Gram-negative enteric bacilli from clinical and nonclinical specimens. For the isolation and identification of microorganisms from fecal specimens. For the isolation of Salmonella, Shigella, and other nonlactose- and nonsucrose-fermenting microorganisms. Nonlactose/nonsucrose-fermenting microorganisms appear as colorless or blue colonies. Lactose/sucrose-fermenting microorganisms, such as coliform bacteria, appear as yellow-opaque white colonies surrounded by a zone of precipitated deoxycholate.

BCP MS G Agar

See: Bromocresol Purple Milk Solids Glucose Agar

BCYE Agar with Cysteine

(BCYE Alpha Base)

(Buffered Charcoal Yeast Extract Agar)

Composition per liter:

Agar

15.0g

Yeast extract

10.0g

ACES buffer (2-[(2-amino-2-oxoethyl)-amino]-ethane sulfonic acid)

10.0g

Charcoal, activated

2.0g

α-Ketoglutarate

1.0g

L-Cysteine·HCl·H2O

0.4g

Fe4(P2O7)3·9H2O

0.25g

L-Cysteine solution

4.0mL

pH 6.9 ± 0.2 at 25°C

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

L-Cysteine Solution:

Composition per 10.0mL:

L-cysteine·HCl·H2O

0.4g

Preparation of L-Cysteine Solution: Add L-cysteine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except L-cysteine solution, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust medium to pH 6.9 with 1N KOH. Heat gently and bring to boil for 1 min. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Aseptically add 4.0mL of L-cysteine solution. Mix thoroughly. Pour into sterile Petri dishes with constant agitation to keep charcoal in suspension.

Use: For the isolation, cultivation, and maintenance of Legionella pneumophila and other Legionella species from environmental and clinical specimens.

BCYE Differential Agar

(Buffered Charcoal Yeast Extract Differential Agar)

Composition per liter:

Agar

15.0g

Yeast extract

10.0g

ACES buffer (2-[(2-amino-2-oxoethyl)-amino]-ethane sulfonic acid)

10.0g

Charcoal, activated

2.0g

α-Ketoglutarate

1.0g

L-Cysteine·HCl·H2O

0.4g

Fe4(P2O7)3·9H2O

0.25g

Bromcresol Purple

0.01g

Bromthymol Blue

0.01g

pH 6.9 ± 0.2 at 25°C

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

Preparation of Medium: Add components, except L-cysteine·HCl·H2O, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust medium to pH 6.9 with 1N KOH. Heat gently and bring to boiling for 1 min. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Add 4.0mL of a 10% solution of L-cysteine·HCl·H2O that has been filter sterilized. Mix thoroughly. Pour into sterile Petri dishes with constant agitation to keep charcoal in suspension.

Use: For the isolation, cultivation, and maintenance of Legionella pneumophila and other Legionella species from environmental and clinical specimens. For the presumptive differential identification of Legionella species based on colony color and morphology. Legionella pneumophila appears as light blue/green colonies. Legionella micdadei appears as blue/gray or dark blue colonies.

BCYE Medium, Diphasic Blood Culture

(Buffered Charcoal Yeast Extract Medium, Diphasic Blood Culture)

Composition per liter:

Agar phase

1.0L

Broth phase

1.0L

pH 6.9 ± 0.2 at 25°C

Agar Phase:

Composition per liter:

Agar

20.0g

ACES buffer (2-[(2-amino-2-oxoethyl)-amino]-ethane sulfonic acid)

10.0g

Yeast extract

10.0g

Charcoal, activated, acid washed

4.0g

KOH

2.8g

α-Ketoglutarate

1.0g

L-Cysteine·HCl·H2O solution

10.0mL

Fe4(P2O7)3·9H2O solution

10.0mL

L-Cysteine·HCl·H2O Solution:

Composition per 10.0mL:

L-Cysteine·HCl·H2O

0.4g

Preparation of L-Cysteine·HCl·H2O Solution: Add L-cysteine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Fe4(P2O7)3·9H2O Solution:

Composition per 10.0mL:

Fe4(P2O7)3·9H2O

0.25g

Preparation of Fe4(P2O7)3·9H2O Solution: Add Fe4(P2O7)3·9H2O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Agar Phase: Add components, except L-cysteine·HCl·H2O solution and Fe4(P2O7)3 solution, to distilled/deionized water and bring volume to 980.0mL. Mix thoroughly. Adjust medium to pH 6.9 with 1N KOH. Heat gently and bring to boiling for 1 min. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°– 55°C. Aseptically add the L-cysteine·HCl·H2O solution and Fe4(P2O7)3·9H2O solution. Mix thoroughly.

Broth Phase:

Composition per liter:

ACES buffer (2-[(2-amino-2-oxoethyl)-amino]-ethane sulfonic acid)

10.0g

Yeast extract

10.0g

Charcoal, activated, acid washed

4.0g

KOH

2.4g

α-Ketoglutarate

1.0g

Sodium polyaneolsulfonate

0.3g

L-Cysteine·HCl·H2O solution

10.0mL

Fe4(P2O7)3·9H2O solution

10.0mL

L-Cysteine·HCl·H2O Solution:

Composition per 10.0mL:

L-Cysteine·HCl·H2O

0.4g

Preparation of L-Cysteine·HCl·H2O Solution: Add L-cysteine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Fe4(P2O7)3·9H2O Solution:

Composition per 10.0mL:

Fe4(P2O7)3·9H2O

0.25g

Preparation of Fe4(P2O7)3·9H2O Solution: Add Fe4(P2O7)3·9H2O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Broth Phase: Add components, except L-cysteine·HCl·H2O solution and Fe4(P2O7)3 solution, to distilled/deionized water and bring volume to 980.0mL. Mix thoroughly. Adjust medium to pH 6.9 with 1N KOH. Heat gently and bring to boiling for 1 min. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50–55°C. Aseptically add the cysteine·HCl·H2O solution and Fe4(P2O7)3·9H2O solution. Mix thoroughly.

Preparation of Medium: Aseptically distribute cooled sterile agar phase into sterile blood culture bottles in 100.0mL volumes. Allow bottles to cool in a slanted position. Aseptically add 50.0mL of sterile broth phase to each blood culture bottle.

Use: For the isolation and cultivation of Legionella pneumophila and other Legionella species from blood samples.

BCYE Selective Agar with CCVC

(Buffered Charcoal Yeast Extract

Selective Agar with Cephalothin,

Colistin, Vancomycin, and Cycloheximide)

Composition per 1014.0mL:

Agar

15.0g

Yeast extract

10.0g

ACES buffer (2-[(2-amino-2-oxoethyl)-amino]-ethane sulfonic acid)

10.0g

Charcoal, activated

2.0g

α-Ketoglutarate

1.0g

Fe4(P2O7)3·9H2O

0.25g

Antibiotic solution

10.0mL

Cysteine·HCl·H2O solution

4.0mL

pH 6.9 ± 0.2 at 25°C

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

L-Cysteine·HCl·H2O Solution:

Composition per 10.0mL:

L-Cysteine·HCl·H2O

1.0g

Preparation of L-Cysteine·HCl·H2O Solution: Add Lcysteine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Antibiotic Solution:

Composition per 10.0mL:

Cycloheximide

80.0mg

Colistin

16.0mg

Cephalothin

4.0mg

Vancomycin

0.5mg

Preparation of Antibiotic Solution: Add components to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Caution: Cycloheximide is toxic. Avoid skin contact or aerosol formation and inhalation.

Preparation of Medium: Add components, except L-cysteine and antibiotic solutions, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust medium to pH 6.9 with 1N KOH. Heat gently and bring to boil for 1 min. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Add 4.0mL of L-cysteine·HCl·H2O solution and 10.0mL of sterile antibiotic solution. Mix thoroughly. Pour into sterile Petri dishes with constant agitation to keep charcoal in suspension.

Use: For the isolation, cultivation, and maintenance of Legionella pneumophila and other Legionella species from environmental and clinical specimens. For the selective recovery of Legionella pneumophila while reducing contaminating microorganisms from environmental water samples.

BCYE Selective Agar with GPVA

(Buffered Charcoal Yeast Extract Selective Agar with

Glycine, Polymyxin B, Vancomycin, and Anisomycin)

Composition per 1014.0mL:

Agar

15.0g

Yeast extract

10.0g

ACES buffer (2-[(2-amino-2-oxoethyl)-amino]-ethane sulfonic acid)

10.0g

Charcoal, activated

2.0g

α-Ketoglutarate

1.0g

Fe4(P2O7)3·9H2O

0.25g

Antibiotic solution

10.0mL

L-Cysteine·HCl·H2O solution

4.0mL

pH 6.9 ± 0.2 at 25°C

L-Cysteine·HCl·H2O Solution:

Composition per 10.0mL:

L-Cysteine·HCl·H2O

1.0g

Preparation of L-Cysteine·HCl·H2O Solution: Add Lcysteine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Antibiotic Solution:

Composition per 10.0mL:

Glycine

3.0g

Anisomycin

0.08g

Vancomycin

5.0mg

Polymyxin B

100,000U

Preparation of Antibiotic Solution: Add components to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except L-cysteine·HCl·H2O solution and antibiotic solution, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust medium to pH 6.9 with 1N KOH. Heat gently and bring to boil for 1 min. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Add 4.0mL of Lcysteine·HCl·H2O solution and 10.0mL of sterile antibiotic solution. Mix thoroughly. Pour into sterile Petri dishes with constant agitation to keep charcoal in suspension.

Use: For the isolation, cultivation, and maintenance of Legionella pneumophila and other Legionella species from environmental and clinical specimens. For the selective recovery of Legionella pneumophila while reducing contaminating microorganisms from potable water samples.

BCYE Selective Agar with GVPC

(Buffered Charcoal Yeast Extract Selective Agar

with Glycine, Vancomycin, Polymyxin B, and Cycloheximide)

Composition per 1014.0mL:

Agar

15.0g

Yeast extract

10.0g

ACES buffer (2-[(2-amino-2-oxoethyl)-amino]-ethane sulfonic acid)

10.0g

Charcoal, activated

2.0g

α-Ketoglutarate

1.0g

Fe4(P2O7)3·9H2O

0.25g

Antibiotic solution

10.0mL

L-Cysteine·HCl·H2O solution

4.0mL

pH 6.9 ± 0.2 at 25°C

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

L-Cysteine·HCl·H2O Solution:

Composition per 10.0mL:

L-Cysteine·HCl·H2O

1.0g

Preparation of L-Cysteine·HCl·H2O Solution: Add Lcysteine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Antibiotic Solution:

Composition per 10.0mL:

Glycine

3.0g

Cycloheximide

0.08g

Vancomycin

1.0mg

Polymyxin B

79,200U

Preparation of Antibiotic Solution: Add components to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Caution: Cycloheximide is toxic. Avoid skin contact or aerosol formation and inhalation.

Preparation of Medium: Add components, except L-cysteine·HCl·H2O solution and antibiotic solution, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust medium to pH 6.9 with 1N KOH. Heat gently and bring to boil for 1 min. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Add 4.0mL of L-cysteine·HCl·H2O solution and 10.0mL of sterile antibiotic solution. Mix thoroughly. Pour into sterile Petri dishes with constant agitation to keep charcoal in suspension.

Use: For the isolation, cultivation, and maintenance of Legionella pneumophila and other Legionella species from environmental and clinical specimens. For the selective recovery of Legionella pneumo- phila while reducing contaminating microorganisms from potable water samples.

BCYE Selective Agar with PAC

(Buffered Charcoal Yeast Extract Selective Agar

with Polymyxin B, Anisomycin, and Cefamandole)

Composition per 1014.0mL:

Agar

15.0g

Yeast extract

10.0g

ACES buffer (2-[(2-amino-2-oxoethyl)-amino]-ethane sulfonic acid)

10.0g

Charcoal, activated

2.0g

α-Ketoglutarate

1.0g

Fe4(P2O7)3·9H2O

0.25g

Antibiotic solution

10.0mL

L-Cysteine·HCl·H2O solution

4.0mL

pH 6.9 ± 0.2 at 25°C

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

L-Cysteine·HCl·H2O Solution:

Composition per 10.0mL:

L-Cysteine·HCl·H2O

1.0g

Preparation of L-Cysteine·HCl·H2O Solution: Add Lcysteine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Antibiotic Solution:

Composition per 10.0mL:

Polymyxin B

80,000 U

Anisomycin

80.0mg

Cefamandole

2.0mg

Preparation of Antibiotic Solution: Add components to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except L-cysteine·HCl·H2O solution and antibiotic solution, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust medium to pH 6.9 with 1N KOH. Heat gently and bring to boiling for 1 min. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Add 4.0mL of L-cysteine·HCl·H2O solution and 10.0mL of sterile antibiotic solution. Mix thoroughly. Pour into sterile Petri dishes with constant agitation to keep charcoal in suspension.

Use: For the isolation, cultivation, and maintenance of Legionella pneumophila and other Legionella species from environmental and clinical specimens. For the selective recovery of Legionella pneumophila while reducing contaminating microorganisms from potable water samples.

BCYE Selective Agar with PAV

(Buffered Charcoal Yeast Extract Selective Agar with

Polymyxin B, Anisomicin, and Vancomycin)

(Wadowsky–Yee Medium)

Composition per 1014.0mL:

Agar

15.0g

Yeast extract

10.0g

ACES buffer (2-[(2-amino-2-oxoethyl)-amino]-ethane sulfonic acid)

10.0g

Charcoal, activated

2.0g

α-Ketoglutarate

1.0g

Fe4(P2O7)3·9H2O

0.25g

Antibiotic solution

10.0mL

L-Cysteine·HCl·H2O solution

4.0mL

pH 6.9 ± 0.2 at 25°C

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

L-Cysteine·HCl·H2O Solution:

Composition per 10.0mL:

L-Cysteine·HCl·H2O

1.0g

Preparation of L-Cysteine·HCl·H2O Solution: Add Lcysteine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Antibiotic Solution:

Composition per 10.0mL:

Anisomycin

80.0mg

Vancomycin

0.5mg

Polymyxin B

40,000 U

Preparation of Antibiotic Solution: Add components to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except L-cysteine and antibiotic solution, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust medium to pH 6.9 with 1N KOH. Heat gently and bring to boil for 1 min. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Add 4.0mL of L-cysteine·HCl·H2O solution and 10.0mL of sterile antibiotic solution. Mix thoroughly. Pour into sterile Petri dishes with constant agitation to keep charcoal in suspension.

Use: For the isolation, cultivation, and maintenance of Legionella pneumophila and other Legionella species from environmental and clinical specimens. For the selective recovery of Legionella pneumophila while reducing contaminating microorganisms from potable water samples.

BCYEα Agar, Modified

See: Legionella Agar Base

BCYEα with Alb

(Buffered Charcoal Yeast Extract Agar with Albumin)

Composition per liter:

Agar

15.0g

Yeast extract

10.0g

ACES buffer (2-[(2-amino-2-oxoethyl)-amino]-ethane sulfonic acid)

10.0g

Charcoal, activated

2.0g

α-Ketoglutarate

1.0g

Bovine serum albumin solution

10.0mL

L-Cysteine·HCl·H2O solution

10.0mL

Fe4(P2O7)3·9H2O solution

10.0mL

pH 6.9 ± 0.2 at 25°C

Bovine Serum Albumin Solution:

Composition per 10.0mL:

Bovine serum albumin

0.1g

Preparation of Bovine Serum Albumin Solution: Add bovine serum albumin to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

L-Cysteine·HCl·H2O Solution:

Composition per 10.0mL:

L-Cysteine·HCl·H2O

0.4g

Preparation of L-Cysteine·HCl·H2O Solution: Add L-cysteine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Fe4(P2O7)3·9H2O Solution:

Composition per 10.0mL:

Fe4(P2O7)3·9H2O

0.25g

Preparation of Fe4(P2O7)3·9H2O Solution: Add Fe4(P2O7)3·9H2O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components—except Fe4(P2O7)3·9H2O solution, L-cysteine·HCl·H2O solution, and bovine serum albumin solution—to distilled/deionized water and bring volume to 970.0mL. Mix thoroughly. Adjust medium to pH 6.9 with 1N KOH. Heat gently and bring to boiling for 1 min. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Aseptically add 10.0mL of sterile bovine serum albumin solution, the Fe4(P2O7)3·9H2O solution, and the L-cysteine·HCl·H2O solution. Mix thoroughly. Pour into sterile Petri dishes with constant agitation to keep charcoal in suspension.

Use: For the isolation, cultivation, and maintenance of Legionella pneumophila and other Legionella species from environmental and clinical specimens.

BCYEα without L-Cysteine

(Buffered Charcoal Yeast Extract Agar

without L-Cysteine)

Composition per liter:

Agar

15.0g

Yeast extract

10.0g

ACES buffer (2-[(2-amino-2-oxoethyl)-amino]-ethane sulfonic acid)

10.0g

Charcoal, activated

2.0g

α-Ketoglutarate

1.0g

Fe4(P2O7)3·9H2O solution

10.0mL

pH 6.9 ± 0.2 at 25°C

Fe4(P2O7)3·9H2O Solution:

Composition per 10.0mL:

Fe4(P2O7)3·9H2O

0.25g

Preparation of Fe4(P2O7)3·9H2O Solution: Add Fe4(P2O7)3·9H2O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except Fe4(P2O7)3·9H2O solution, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Adjust medium to pH 6.9 with 1N KOH. Heat gently and bring to boiling for 1 min. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Aseptically add 10.0mL of sterile Fe4(P2O7)3·9H2O solution. Mix thoroughly. Pour into sterile Petri dishes with constant agitation to keep charcoal in suspension.

Use: For the isolation, cultivation, and maintenance of Legionella pneumophila and other Legionella species from environmental and clinical specimens.

BCYT

See: Methanosarcina Medium

Bdellovibrio Medium

Composition per Petri dish:

Base layer agar

10.0mL

Semisolid agar

10.0mL

Host medium

1.0mL

Host Medium:

Composition per liter:

Yeast extract

3.0g

Peptone

0.6g

Preparation of Host Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.2. Distribute into tubes in 10.0mL volumes. Autoclave for 15 min at 15 psi pressure–121°C.

Base Layer Agar:

Composition per liter:

Agar

19.0g

Yeast extract

3.0g

Peptone

0.6g

Preparation of Base Layer Agar: 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 in 10.0mL volumes. Autoclave for 15 min at 15 psi pressure–121°C.

Semisolid Agar:

Composition per liter:

Agar

6.0g

Yeast extract

3.0g

Peptone

0.6g

Preparation of Semisolid Agar: 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 in 10.0mL volumes. Autoclave for 15 min at 15 psi pressure–121°C.

Preparation of Medium: Inoculate appropriate bacterial host into 10.0mL of host medium. Hosts include Erwinia amylovora, Escherichia coli, Serratia marcescens, or Pseudomonas putida. Incubate host culture for 24–48 hr at 30°C. Melt the base layer agar and semisolid agar. Pour the base layer agar into a sterile Petri dish. Allow base layer agar to solidify. Cool the semisolid agar to 40°–45°C. Add 1.0mL of the previously grown host culture. Mix thoroughly. Pour over the solidified base layer agar.

Use: For the cultivation of Bdellovibrio bacteriovorus and Bdellovibrio starrii.

B.D.G. Broth, Hajna

Composition per liter:

Tryptose

20.0g

Glucose

5.0g

NaCl

5.0g

K2HPO4

4.0g

KH2PO4

1.5g

Sodium deoxycholate

0.1g

pH 7.0 ± 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 selective enrichment and cultivation of enteric bacilli from food and in treated drinking water.

Bean Agar

Composition per liter:

Dry white beans

250.0g

Agar

20.0g

Preparation of Medium: Soak beans in 500.0mL of distilled/deionized water for 12 hr. Autoclave for 20 min at 15 psi pressure– 121°C. Filter broth through cotton. Bring volume of filtrate to 1.0L with distilled/deionized water. Add 20.0g of agar to the filtrate. 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 Arthroderma melis and Rhynchosporium secalis.

Beef Extract Agar

Composition per liter:

Agar

15.0g

Peptone

5.0g

Beef extract

3.0g

pH 7.4 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Heat gently and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into Petri dishes or leave in tubes.

Use: For the cultivation and maintenance of a wide variety of micro-organisms. Recommended for the culture of microorganisms from milk and water.

Beef Extract Agar

(ATCC Medium 225)

Composition per liter:

Agar

25.0g

Beef extract

10.0g

Peptone

10.0g

NaCl

5.0g

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Heat gently and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into Petri dishes or leave in tubes.

Use: For the cultivation and maintenance of a wide variety of micro-organisms, including Alcaligenes species, Pseudomonas aeruginosa, and Bacillus sphaericus.

Beef Extract Agar, HiVeg

Composition per liter:

Agar

15.0g

Plant peptone

10.0g

NaCl

5.0g

Plant extract

3.0g

pH 7.6 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Hi-Media.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Heat gently and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into Petri dishes or leave in tubes.

Use: For the cultivation and maintenance of a wide variety of micro-organisms, including Alcaligenes species, Pseudomonas aeruginosa, and Bacillus sphaericus.

Beef Extract Broth

Composition per liter:

Peptone

5.0g

Beef extract

3.0g

pH 7.4 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Heat gently and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the cultivation and maintenance of a wide variety of micro-organisms. Recommended for the culture of microorganisms from milk and water.

Beef Extract Broth

(ATCC Medium 225)

Composition per liter:

Beef extract

10.0g

Peptone

10.0g

NaCl

5.0g

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Heat gently and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the cultivation of a wide variety of microorganisms, including Alcaligenes species, Pseudomonas aeruginosa, and Bacillus sphaericus.

Beef Extract Broth, HiVeg

Composition per liter:

Plant peptone

10.0g

NaCl

5.0g

Plant extract

3.0g

pH 7.2 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Hi-Media.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Heat gently and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the cultivation and maintenance of a wide variety of micro-organisms. Recommended for the culture of microorganisms from milk and water.

Beef Extract Peptone Serum Medium

Composition per liter:

Agar

25.0g

Beef extract

10.0g

Peptone

10.0g

NaCl

1.0g

Bovine serum

50.0mL

pH 8.5 ± 0.2 at 25°C

Preparation of Medium: Add components, except bovine serum, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Adjust pH to 8.5. Heat gently and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Aseptically add 50.0mL of sterile bovine serum. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation and maintenance of Serratia marcescens.

Beef Extract V

Composition per liter:

Beef extract

24.0g

pH 9.0 at 25°C

Preparation of Medium: Add component to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 9.0 with NaOH. Autoclave for 15 min at 15 psi pressure—118°–121°C.

Use: For use in the elution of viruses that have been adsorbed onto filters during the filtration of water and wastewater samples.

Beef Extract with Sodium Chloride

Composition per liter:

Beef extract

10.0g

NaCl

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 Bacillus megaterium.

Beef Infusion Agar

Composition per liter:

Ground defatted beef

453.6g

Agar

20.0g

Peptone

10.0g

NaCl

5.0g

pH 7.6 ± 0.2 at 25°C

Preparation of Medium: Add ground beef to 1.0L of distilled/deionized water. Let stand overnight at 4°C. Gently heat and bring to 80°–90°C for 60 min. Let stand for 2 hr. Filter through muslin. To filtrate, add pep-tone and salt. Mix thoroughly. Adjust pH to 7.6 with 4% NaOH. Filter through Whatman #1 filter paper. Bring volume of filtrate to 1.0L. Add agar. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation of a variety of microorganisms.

Beef Infusion Broth

Composition per liter:

Ground beef, defatted

453.6g

Peptone

10.0g

NaCl

5.0g

pH 7.6 ± 0.2 at 25°C

Preparation of Medium: Add ground beef to 1.0L of distilled/deionized water. Let stand overnight at 4°C. Gently heat and bring to 80°–90°C for 60 min. Let stand for 2 hr. Filter through muslin. To filtrate add peptone and salt. Mix thoroughly. Adjust pH to 7.6 with 4% NaOH. Filter through Whatman #1 filter paper. Bring volume of filtrate to 1.0L. Add agar. 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 a variety of microorganisms.

Beef Liver Medium for Anaerobes

Composition per liter:

Beef liver, minced

500.0g

Peptone

10.0g

K2HPO4

1.0g

pH 8.0 ± 0.2 at 25°C

Preparation of Medium: Add beef liver to 1.0L of tap water. Soak for 12–24 hr at 4°C. Skim fat off top. Autoclave for 10 min at 15 psi pressure–121°C. Filter through cheesecloth. Save meat. To filtrate, add pep-tone and K2HPO4. Adjust pH to 8.0. Filter through paper. Add tap water and bring volume to 1.0L. Add a small amount of CaCO3 to a flask or test tube. Add 0.5 inch of reserved liver. Cover meat with 2 inches of broth. Cap tubes and autoclave for 15 min at 15 psi pressure–121°C.

Use: For the cultivation and maintenance of a variety of Clostridium species.

Beggiatoa Agar

Composition per 1010.0mL:

Agar

10.0g

Sodium acetate

0.5g

Pancreatic digest of gelatin

.0.31g

Beef extract

0.19g

NH4Cl

0.45mg

MgSO4·7H2O

0.2mg

K2HPO4

0.1mg

CaSO4 (saturated solution)

20.0mL

Catalase solution

10.0mL

Trace elements solution

5.0mL

pH 7.4 ± 0.2 at 25°C

Catalase Solution:

Composition per 10.0mL:

Catalase

15,000–35,000U

Preparation of Catalase Solution: Add catalase to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Trace Elements Solution:

Composition per liter:

FeSO4·7H2O

0.7g

EDTA

0.2g

ZnSO4·7H2O

0.01g

MnSO4·4H2O

0.002g

H3BO3

10.0mg

CO(NO3)2

1.0mg

Na2MoO4·2H2O

1.0mg

CuSO4·5H2O

5.0μg

Preparation of Trace Elements Solution: Add FeSO4·7H2O to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized water and bring volume to 1.0L. Add remaining components. Mix thoroughly.

Preparation of Medium: Add components, except catalase solution, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Adjust pH to 7.4. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically add 10.0mL of sterile catalase solution (freshly prepared). Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation and maintenance of Beggiatoa alba.

Beggiatoa and Thiothrix Medium

Composition per liter:

CaSO4·2H2O (saturated solution)

20.0mL

NH4Cl (4% solution)

5.0mL

Trace elements

5.0mL

K2HPO4 (1% solution)

1.0mL

MgSO4·7H2O (1% solution)

1.0mL

Trace Elements:

Composition per liter:

EDTA solution

20.0mL

Co(NO3)2 (0.01% solution)

10.0mL

CuSO4·5H2O (0.00005% solution)

10.0mL

H3BO3 (0.1% solution)

10.0mL

MnSO4·4H2O (0.02% solution)

10.0mL

Na2MoO4·2H2O (0.01% solution)

10.0mL

ZnSO4·7H2O (0.1% solution)

10.0mL

Preparation of Trace Elements: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

EDTA Solution:

Composition per 100.0mL:

FeSO4

7.0g

EDTA

2.0g

HCl, concentrated

1.0mL

Preparation of EDTA Solution: Add EDTA and FeSO4 to concentrated HCl. Mix thoroughly. Carefully add to distilled/deionized water and bring volume to 100.0mL.

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 Beggiatoa species and myxotrophic Thiothrix species.

Beggiatoa Broth

Composition per 1010.0mL:

Sodium acetate

0.5g

Pancreatic digest of gelatin

.0.31g

Beef extract

0.19g

NH4Cl

0.45mg

MgSO4·7H2O

0.2mg

K2HPO4

0.1mg

CaSO4 (saturated solution)

20.0mL

Catalase solution

10.0mL

Trace elements solution

5.0mL

pH 7.4 ± 0.2 at 25°C

Catalase Solution:

Composition per 10.0mL:

Catalase

15,000–35,000U

Preparation of Catalase Solution: Add catalase to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Trace Elements Solution:

Composition per liter:

FeSO4·7H2O

0.7g

EDTA

0.2g

ZnSO4·7H2O

0.01g

MnSO4·4H2O

0.002g

H3BO3

1.0mg

Na2MoO4·2H2O

1.0mg

CuSO4·5H2O

5.0μg

Preparation of Trace Elements Solution: Add FeSO4·7H2O to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized water and bring volume to 1.0L. Add remaining components. Mix thoroughly.

Preparation of Medium: Add components, except catalase solution, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.4. Autoclave for 15 min at 15 psi pressure– 121°C. Aseptically add 10.0mL of sterile catalase solution (freshly prepared). Mix thoroughly. Aseptically distribute into sterile tubes or flasks.

Use: For the cultivation of Beggiatoa alba.

Beggiatoa Medium

(ATCC Medium 138)

Composition per liter:

Yeast extract

2.0g

Agar

2.0g

Sodium acetate

0.5g

CaCl2

0.1g

Catalase

10,000U

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components, except catalase, to tap water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add 10,000 units of sterile catalase.

Use: For the cultivation and maintenance of Beggiatoa alba and Vitreoscilla species.

Beggiatoa Medium

(ATCC Medium 1193)

Composition per liter:

Sodium sulfide

0.5g

Sodium acetate

0.01g

Yeast extract

0.01g

Nutrient broth

0.01g

pH 7.5 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Distribute into tubes or flasks.

Use: For the cultivation of Beggiatoa alba.

Beijerinckia Agar

Composition per liter:

Agar

15.0g

K2HPO4

0.8g

KH2PO4

0.2g

MgSO4·7H2O

0.1g

FeSO4·7H2O

20.0mg

Na2MoO4·2H2O

5.0mg

ZnSO4·6H2O

5.0mg

CuSO4·6H2O

4.0mg

MnSO4·6H2O

2.0mg

Glucose solution

50.0mL

pH 6.5 ± 0.2 at 25°C

Glucose Solution:

Composition per 50.0mL:

D-Glucose

10.0g

Preparation of Glucose Solution: Add glucose to distilled/deionized 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 glucose solution, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically add 10.0mL of sterile glucose solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes

Use: For the cultivation and maintenance of Beijerinckia derxii, Beijerinckia fluminensis, Beijerinckia indica, Beijerinckia mobilis, Beijerinckia species, and Clostridium barkeri.

Beijerinckia Medium

Composition per liter:

Glucose

20.0g

KH2PO4

1.0g

MgSO4·7H2O

0.5g

pH 5.0 ± 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 Beijerinckia species.

Beijerinckia Medium

Composition per liter:

Glucose

20.0g

K2HPO4

0.8g

MgSO4·7H2O

0.5g

KH2PO4

0.2g

CaCl2

0.05g

FeCl3·6H2O

0.025g

Na2MoO4·2H2O

5.0mg

pH 6.9 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the isolation and cultivation of Beijerinckia species.

Beijerinckia Medium

Composition per liter:

Sucrose

20.0g

Agar

15.0g

KH2PO4

0.8g

MgSO4·7H2O

0.5g

K2HPO4

0.2g

FeCl3·6H2O

0.1g

Na2MoO4·2H2O

5.0mg

pH 6.5 ± 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 or flasks. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the isolation and cultivation of Beijerinckia species.

Beijerinckia Medium, Modified

Composition per liter:

Agar

15.0g

Glucose

10.0g

K2HPO4

0.8g

KH2PO4

0.2g

MgSO4·7H2O

0.1g

FeSO4·7H2O

20.0mg

MnSO4·H2O

1.3mg

ZnSO4·7H2O

5.0mg

CuSO4·5H2O

4.0mg

Na2MoO4·2H2O

5.0mg

pH 6.5 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.

Use: For the isolation and cultivation of Beijerinckia derxii, Beijerinckia fluminensis, Beijerinckia indica, and Beijerinckia mobilis.

Beijerinck’s Thiobacillus Medium

Composition per liter:

Noble agar

20.0g

Na2HPO4

0.2g

MgCl2

0.1g

NH4Cl

0.1g

Na2S2O3 solution

100.0mL

NaHCO3 solution

10.0mL

pH 7.0–7.2 at 25°C

Na2S2O3 Solution:

Composition per 100.0mL:

Na2S2O3

5.0g

Preparation of Na2S2O3 Solution: Add Na2S2O3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

NaHCO3 Solution:

Composition per 10.0mL:

NaHCO3

1.0g

Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except Na2S2O3 solution and NaHCO3 solution, to distilled/deionized water and bring volume to 890.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically add 100.0mL of sterile Na2S2O3 solution and 10.0mL of sterile NaHCO3 solution. Mix thoroughly. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation and maintenance of Thiobacillus thermophilica.

Bennett’s Agar

Composition per liter:

Agar

15.0g

Glucose

10.0g

N-Z amine, type A

2.0g

Beef extract

1.0g

Yeast extract

1.0g

pH 7.3 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.3. Gently heat and bring to boiling. Distribute into tubes or flasks. Auto-clave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation and maintenance of Actinomadura umbrina, Micromonospora purpurea, Microtetraspora helvata, Nocardia salmonicolor, and Streptomyces species.

Bennett’s Agar with Maltose

Composition per liter:

Agar

15.0g

Maltose, technical

10.0g

N-Z amine, type A

2.0g

Beef extract

1.0g

Yeast extract

1.0g

pH 7.3 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.3. Gently heat and bring to boiling. Distribute into tubes or flasks. Auto-clave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation and maintenance of Streptomyces species.

Bennett’s Agar with Sucrose

Composition per liter:

Agar

15.0g

Sucrose

10.0g

N-Z amine, type A

2.0g

Beef extract

1.0g

Yeast extract

1.0g

pH 7.3 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.3. Gently heat and bring to boiling. Distribute into tubes or flasks. Auto-clave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation and maintenance of Actinomadura madurae, Excellospora viridilutea, Geodermatophilus obscurus, Intrasporangium calvum, Kibdelosporangium aridum, Microbispora thermodiastatica, Micromonospora coerulea, Micromonospora echinospora, Micromonospora purpureochromogenes, Micromonospora rosaria, Microtetraspora flexuosa, Promicromonospora enterophila, Saccharomonospora glauca, Streptomyces cacaoi, Thermoactinomyces dichotomicus, Thermoactinomyces glaucus, and Thermomonospora chromogena.

Bennet’s HiVeg Agar

Composition per liter:

Agar

15.0g

Glucose

10.0g

Plant hydrolysate

2.0g

Plant extract

1.0g

Yeast extract

1.0g

pH 7.3 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Hi-Media.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.3. Gently heat and bring to boiling. Distribute into tubes or flasks. Auto-clave for 10 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation and maintenance of Actinomadura umbrina, Micromonospora purpurea, Microtetraspora helvata, Nocardia salmonicolor, and Streptomyces species.

Bennett’s Medium

Composition per liter:

Agar

15.0g

Glucose

10.0g

Pancreatic digest of casein

2.0g

Yeast extract

1.0g

Beef extract

1.0g

pH 7.0 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Heat gently 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 soil microorganisms, such as Streptomyces species, Nocardia species, Flexibacter species, Micromonospora species, and others.

Bennett’s Modified Agar Medium

Composition per liter:

Meer agar (washed agar)

20.0g

Dextrin

10.0g

Pancreatic digest of casein

2.0g

Yeast extract

1.0g

Beef extract

1.0g

CoCl2·6H2O

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. Heat gently 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 species.

Benzene Sulfonate Medium

Composition per liter:

Agar

15.0g

Sodium benzene sulfonate

1.0g

(NH4)2SO4

1.0g

K2HPO4

0.7g

KH2PO4

0.3g

MgSO4·7H2O

0.2g

CaCl2

10.0mg

FeSO4·7H2O

5.0mg

ZnSO4·7H2O

70.0μg

CuSO4

50.0μg

H3BO3

10.0μg

MoO3·2H2O

10.0μg

MnSO4·5H2O

2.0μg

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Heat gently to boil ing. 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 Comamonas testosteroni.

Benzoate Medium

Composition per liter:

Noble agar

20.0g

NaCl

5.0g

(NH4)2HPO4

3.0g

Sodium benzoate

3.0g

KH2PO4

1.2g

Yeast extract

0.5g

MgSO4·7H2O

0.2g

Benzoate solution

25.0mL

Benzoate Solution:

Composition per 25.0mL:

Sodium benzoate

3.0g

Preparation of Benzoate Solution: Add sodium benzoate to distilled/deionized water and bring volume to 25.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components except benzoate solution to distilled/deionized water and bring volume to 975.0mL. Mix thoroughly. Heat gently to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add 25.0mL sterile benzoate solution. Mix thoroughly and pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation of Pseudomonas putida and other microorganisms which can utilize benzoate as a carbon source.

Benzoate Medium II

Composition per 1.5L: Noble agar

30.0g

(NH4)2HPO4

3.0g

NaCl

1.67g

KH2PO4

1.2g

Yeast extract

0.5g

MgSO4·7H2O

0.2g

FeSO4·7H2O

0.1g

Benzoate solution

25.0mL

Benzoate Solution:

Composition per 25.0mL:

Sodium benzoate

1.0g

Preparation of Benzoate Solution: Add sodium benzoate to distilled/deionized water and bring volume to 25.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except agar and sodium benzoate, to distilled/deionized water and bring volume to 600.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. In a separate flask, add agar to distilled/deionized water and bring volume to 375.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically combine the two autoclave-sterilized solutions. Mix thoroughly. Aseptically add the sterile benzoate solution. Mix thoroughly. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation of Pseudomonas putida and other microorganisms that can utilize benzoate as a carbon source.

Benzoate Minimal Salts Medium

Composition per liter:

K2HPO4

10.0g

NaNH4HPO4·4H2O

3.5g

MgSO4·7H2O

0.2g

Citric acid, anhydrous

0.2g

Benzoate solution

25.0mL

pH 7.0 ± 0.2 at 25°C

Benzoate Solution:

Composition per 25.0mL:

Sodium benzoate

2.5g

Preparation of Benzoate Solution: Add sodium benzoate to distilled/deionized water and bring volume to 25.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 975.0mL. Mix thoroughly. Adjust pH to 7.0. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°C. Aseptically add 25.0mL of sterile benzoate solution. Mix thoroughly. Aseptically distribute into sterile tubes or flasks.

Use: For the cultivation of microorganisms that can utilize benzoate as a carbon source.

Benzoate Nitrate Salts Medium

(BNS)

Composition per liter:

Solution A

700.0mL

Solution B

300.0mL

pH 8.2 ± 0.2 at 25°C

Solution A:

Composition per 700.0mL:

KNO3

2.0g

Sodium benzoate

1.0g

NH4Cl

0.3g

Phosphate buffer solution

200.0mL

Phosphate Buffer Solution:

Composition per 200.0mL:

K2HPO4

5.12g

KH2PO4

1.5g

Preparation of Phosphate Buffer: Add components to distilled/deionized water and bring volume to 200.0mL. Mix thoroughly. Adjust pH to 9.0 with KOH.

Preparation of Solution A: Add components to distilled/deionized water and bring volume to 700.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature.

Solution B:

Composition per 300.0mL:

MgSO4·7H2O

0.2g

CaCl2

10.0mg

Trace metals solution

1.0mL

Preparation of Solution B: Add components to distilled/deionized water and bring volume to 300.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature.

Trace Metals Solution:

Composition per 300.0mL:

MnSO4·H2O

50.0mg

ZnSO4·7H2O

50.0mg

Co(NO3)2·6H2O

10.0mg

CuSO4

10.0mg

Na2B4O7·10H2O

10.0mg

Na2MoO4·2H2O

0.2mg

Ferric EDTA solution

10.0mL

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

Ferric EDTA Solution:

Composition per 550.0mL:

EDTA

17.9g

FeSO4·7H2O

13.7g

KOH

3.23g

Preparation of Ferric EDTA Solution: Add EDTA and KOH to distilled/deionized water and bring volume to 186.0mL. Mix thoroughly. In a separate flask, add FeSO4·7H2O to distilled/deionized water and bring volume to 364.0mL. Mix thoroughly. Combine the two solutions. Sparge with air overnight to oxidize the Fe2+ to Fe3+. Store in the dark.

Preparation of Medium: Aseptically combine 700.0mL of sterile solution A with 300.0mL of sterile solution B. Adjust pH to 8.2. Aseptically distribute into sterile screw-capped tubes. Fill tubes completely.

Use: For the cultivation of Alcaligenes xylosoxydans.

Betabacterium Medium

Composition per liter:

Pancreatic digest of casein

10.0g

Agar

10.0g

Yeast extract

5.0g

Glucose

5.0g

K2HPO4

2.0g

Liver extract

100.0mL

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add 1 pound of finely ground beef liver to 2.0L of distilled/deionized water. Autoclave for 2.5–3 hr at 15 psi pressure–121°C under flowing steam. The liquid should become fluorescent yellow. Filter through sterile cheesecloth. Save solids and dry at 50°C. Add a few pieces of the dried liver to sterile test tubes or flasks. Prepare basal medium by adding components to distilled/deionized water and bring volume to 1.0L. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically add sterile basal medium to each test tube or flask containing liver. Commercial liver extract may be used at a concentration of 0.1%.

Use: For the growth and maintenance of Lactobacillus species. Beta-bacterium is an archaic name that was used to describe several bacteria as a subgenus of the Lactobacillus group.

BG Sulfa Agar

(Brilliant Green Sulfapyridine Agar)

Composition per liter:

Agar

20.0g

Proteose peptone No. 3

10.0g

Lactose

10.0g

Sucrose

10.0g

NaCl

5.0g

Yeast extract

3.0g

Sodium sulfapyridine

1.0g

Brilliant Green

0.125g

pH 6.9 ± 0.2 at 25°C

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

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Heat gently to boiling. Distribute into tubes or flasks. Autoclave for no longer than 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes if desired.

Use: For the selective isolation of Salmonella species other than Salmonella typhi from food, dairy products, eggs and egg products, and feed. Salmonella appear as red, pink, or white colonies surrounded by zones of bright red.

BG Sulfa HiVeg Agar

(Brilliant Green Sulfa HiVeg Agar)

Composition per liter:

Agar

20.0g

Plant peptone No. 3

10.0g

Lactose

10.0g

Sucrose

10.0g

NaCl

5.0g

Yeast extract

3.0g

Sodium sulphapyridine

1.0g

Phenol Red

0.08g

Brilliant Green

12.5mg

pH 6.9 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Hi-Media.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Heat gently to boiling. Distribute into tubes or flasks. Autoclave for no longer than 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes if desired.

Use: For the selective isolation of Salmonella species other than Salmonella typhi from food, dairy products, eggs and egg products, and feed. Salmonella appear as red, pink, or white colonies surrounded by zones of bright red.

BG 11 Agar

(Medium BG 11 for Cyanobacteria)

Composition per liter:

Agar

10.0g

NaNO3

1.5g

MgSO4·7H2O

0.075g

K2HPO4

0.04g

CaCl2·2H2O

0.036g

Na2CO3

0.02g

Citric acid

6.0mg

Ferric ammonium citrate

6.0mg

Disodium EDTA

1.0mg

Trace metal mix A5

1.0mL

pH 7.1 ± 0.2 at 25°C

Trace Metal Mix A5:

Composition per liter:

H3BO3

2.86g

MnCl2·4H2O

1.81g

Na2MoO4·2H2O

0.39g

ZnSO4·7H2O

0.222g

CuSO4·5H2O

0.079g

Co(NO3)2·6H2O

0.049g

Preparation of Trace Metal Mix A5: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Heat gently to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. For solid medium, pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation and maintenance of a variety of cyanobacteria, including Anabaena species, Calothrix species, Chaemisiphon species, Chorogloeopsis species, Chroococcidiopsis species, Cylindrospermum species, Dermocarpa species, Fischerella species, Gloebacter species, Gloeocapsa species, Gloeothece species, Nostoc species, Oscillatoria species, Phormidium species, Pleurocapsa species, Pseudanabaena species, Scytonema species, Spirulina species, Synechococcus species, and Synechocystis species.

BG 11 Marine Agar

(Medium BG 11 for Marine Cyanobacteria)

Composition per liter:

Agar

10.0g

NaCl

10.0g

NaNO3

1.5g

MgSO4·7H2O

0.075g

K2HPO4

0.04g

CaCl2·2H2O

0.036g

Na2CO3

0.02g

Citric acid

6.0mg

Ferric ammonium citrate

6.0mg

EDTA disodium salt

1.0mg

Vitamin B12 solution

100.0mL

Trace metal mix A5

1.0mL

pH 7.1 ± 0.2 at 25°C

Trace Metal Mix A5:

Composition per liter:

H3BO3

2.86g

MnCl2·4H2O

1.81g

Na2MoO4·2H2O

0.39g

ZnSO4·7H2O

0.222g

CuSO4·5H2O

0.079g

Co(NO3)2·6H2O

0.049g

Preparation of Trace Metal Mix A5: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

Vitamin B12 Solution:

Composition per 100.0mL:

Vitamin B12

1.0μg

Preparation of Vitamin B12 Solution: Add vitamin B12 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except vitamin B12 solution, to distilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Heat gently to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically add 100.0mL of sterile vitamin B12 solution. Mix thoroughly. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation and maintenance of Synechococcus species. For the isolation of cyanobacteria from freshwater habitats.

BG 11 Marine Broth

(Medium BG 11 for Marine Cyanobacteria)

Composition per liter:

NaCl

10.0g

NaNO3

1.5g

MgSO4·7H2O

0.075g

K2HPO4

0.04g

CaCl2·2H2O

0.036g

Na2CO3

0.02g

Citric acid

6.0mg

Ferric ammonium citrate

6.0mg

EDTA disodium salt

1.0mg

Vitamin B12 solution

100.0mL

Trace metal mix A5

1.0mL

pH 7.1 ± 0.2 at 25°C

Trace Metal Mix A5:

Composition per liter:

H3BO3

2.86g

MnCl2·4H2O

1.81g

Na2MoO4·2H2O

0.39g

ZnSO4·7H2O

0.222g

CuSO4·5H2O

0.079g

Co(NO3)2·6H2O

0.049g

Preparation of Trace Metal Mix A5: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

Vitamin B12 Solution:

Composition per 100.0mL:

Vitamin B12

1.0μg

Preparation of Vitamin B12 Solution: Add vitamin B12 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except vitamin B12 solution, to distilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Heat gently to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically add 100.0mL of sterile vitamin B12 solution. Mix thoroughly. Distribute into sterile tubes or flasks.

Use: For the cultivation and maintenance of Synechococcus species. For the isolation of cyanobacteria from freshwater habitats.

BG 11 Medium

(Medium BG 11 for Cyanobacteria)

Composition per liter:

Agar

10.0g

NaNO3

1.5g

MgSO4·7H2O

0.075g

K2HPO4

0.04g

CaCl2·2H2O

0.036g

Na2CO3

0.02g

Citric acid

6.0mg

Ferric ammonium citrate

6.0mg

EDTA disodium salt

1.0mg

Trace metal mix A5

1.0mL

pH 7.1 ± 0.2 at 25°C

Trace Metal Mix A5:

Composition per liter:

H3BO3

2.86g

MnCl2·4H2O

1.81g

Na2MoO4·2H2O

0.39g

ZnSO4·7H2O

0.222g

CuSO4·5H2O

0.079g

Co(NO3)2·6H2O

0.049g

Preparation of Trace Metal Mix A5: Add components to distilled/deionized water and bring volume to 1.0L. 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. 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 Anabaena species, Calothrix species, Chaemisiphon species, Chorogloeopsis species, Chroococcidiopsis species, Crinalium epipsammum, Cylindrospermum species, Dermocarpa species, Fischerella species, Gloebacter violaceus, Gloeocapsa species, Gloeothece species, Hapalosiphon fontinalis, Nostoc species, Oscillatoria species, Phormidium species, Pleurocapsa species, Pseudanabaena species, Scytonema species, Spirulina species, Synechococcus species, Synechocystis species, and Tolypothrix tenuis.

BG 11 Uracil Agar

Composition per liter:

Agar

10.0g

Uracil

2.8g

NaNO3

1.5g

MgSO4·7H2O

0.075g

K2HPO4

0.04g

CaCl2·2H2O

0.036g

Na2CO3

0.02g

Citric acid

6.0mg

Ferric ammonium citrate

6.0mg

EDTA disodium salt

1.0mg

Trace metal mix A5

1.0mL

pH 7.1 ± 0.2 at 25°C

Trace Metal Mix A5:

Composition per liter:

H3BO3

2.86g

MnCl2·4H2O

1.81g

Na2MoO4·2H2O

0.39g

ZnSO4·7H2O

0.222g

CuSO4·5H2O

0.079g

Co(NO3)2·6H2O

0.049g

Preparation of Trace Metal Mix A5: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Heat gently to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes.

Use: For the cultivation and maintenance of Anabaena variabilis.

BG 11 Uracil Broth

Composition per liter:

Uracil

2.8g

NaNO3

1.5g

MgSO4·7H2O

0.075g

K2HPO4

0.04g

CaCl2·2H2O

0.036g

Na2CO3

0.02g

Citric acid

6.0mg

Ferric ammonium citrate

6.0mg

EDTA disodium salt

1.0mg

Trace metal mix A5

1.0mL

pH 7.1 ± 0.2 at 25°C

Trace Metal Mix A5:

Composition per liter:

H3BO3

2.86g

MnCl2·4H2O

1.81g

Na2MoO4·2H2O

0.39g

ZnSO4·7H2O

0.222g

CuSO4·5H2O

0.079g

Co(NO3)2·6H2O

0.049g

Preparation of Trace Metal Mix A5: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Heat gently to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the cultivation and maintenance of Anabena variabilis.

BHI

See : Brain Heart Infusion

BHI Agar

See: Brain Heart Infusion Agar

BHI Broth

See: Brain Heart Infusion Broth

BHI Glucose Medium

Composition per liter:

Agar

12.0g

Pancreatic digest of gelatin

7.25g

Glucose

6.5g

Brain heart, solids from infusion

3.0g

Peptic digest of animal tissue

3.0g

NaCl

2.5g

Na2HPO4

1.25g

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

Use: For the cultivation and maintenance of Actinomadura pelletieri, Actinoplanes missouriensis, Actinoplanes philippinensis, Agromyces ramosus, Corynebacterium minutissimum, Dermatophilus congolensis, Intrasporangium calvum, Mycobacterium diernhoferi, Mycobacterium species, Nocardia asteroides, Nocardia brevicatena, Nocardia calcarea, Nocardia otitidiscaviarum, Pseudonocardia thermophila, Saccharopolyspora rectivirgula, Streptococcus iniae, and Streptococcus pyo-genes.

BHI with Glucose (DSMZ Medium 215b)

Composition per liter:

Pancreatic digest of gelatin

14.5g

Glucose

8.0g

Brain heart, solids from infusion

6.0g

Peptic digest of animal tissue

6.0g

NaCl

5.0g

Na2HPO4

2.5g

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

Use: For the cultivation of Corynebacterium spp., Streptomyces flocculus, Mycobacterium spp., Nocardia spp., Rhodococcus spp., Dermatophilus congolensis, and Gordonia amicalis.

BHI with Glycerol and Reducing Agents

(DSMZ Medium 215c)

Composition per liter:

Pancreatic digest of gelatin

14.5g

Brain heart, solids from infusion

6.0g

Peptic digest of animal tissue

6.0g

NaCl

5.0g

Glucose

3.0g

Na2HPO4

2.5g

Glycerol solution

10.0mL

L-Cysteine·HCl–Na2S solution

10.0mL

pH 7.4 ± 0.2 at 25°C

Glycerol Solution:

Composition per 100.0mL:

Glycerol

87.0g

Preparation of Glycerol Solution: Add glycerol to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly.

L-Cysteine·HCl–Na2S Solution:

Composition per 100.0mL:

L-Cysteine·HCl

2.5g

Na2S·9H2O

2.5g

Preparation of L-Cysteine·HCl–Na2S Solution: Add L-cysteine·HCl to distilled/deionized water and bring volume to 80.0mL. Mix thoroughly. Adjust pH to 11 with NaOH. Add Na2S·9H2O. Mix thoroughly. Bring volume to 100.0mL with distilled/deionized water. Gently heat and bring to boiling under 100% N2. Cool to 25°C under 100% N2. Autoclave for 15 min at 15 psi pressure–121°C.

Preparation of Medium: Add components, except L-cysteine·HCl– Na2S solution, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Gently heat and bring to boiling under 100% N2. Cool to 25°C under 100% N2. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically under 100% N2 add 10.0mL of L-cysteine·HCl–Na2S solution. Mix thoroughly. Aseptically under 100% N2 distribute to tubes. Alternately distribute 10.0mL amounts of the medium without L-cysteine·HCl–Na2S solution to tubes prior to autoclaving. Auto-clave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add 1.0mL of L-cysteine·HCl–Na2S solution to each tube.

Use: For the cultivation of Clostridium sp.

BHI Medium

(DSMZ Medium 215)

Composition per liter:

Pancreatic digest of gelatin

14.5g

Brain heart, solids from infusion

6.0g

Peptic digest of animal tissue

6.0g

NaCl

5.0g

Na2HPO4

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 15 min at 15 psi pressure–121°C.

Use: For the cultivation of Enterococcus hirae, Lodobacter fluviatilis, Yersinia spp., Tatumella ptyseos, Mycobacterium vanbaalenii, Oligella urethralis=Moraxella urethralis, Moraxella (Branhamella) catarrhalis, Campylobacter sputorum, Helcococcus kunzii, Bacillus sporothermodurans, Haemophilus actinomycetemcomitans, Escherichia coli, Pelczaria aurantia, Bacillus spp., Comamonas nitrativorans, Salmonella bongori (Salmonella choleraesuis subsp. bongori), Sphingomonas sanguinis (Sphingomonas sanguis), Arsenophonus nasoniae, Streptococcus orisratti, Listeria spp., Jonesia denitrificans=Listeria denitrificans, Propionibacterium propionicus=Arachnia propionica, Corynebacterium spp., and Nocardiopsis tropica.

BHI/1 Medium

Composition per liter:

Pancreatic digest of gelatin

14.5g

Casein hydrolysate

10.0g

Glucose

8.0g

Brain heart, solids from infusion

6.0g

Peptic digest of animal tissue

6.0g

NaCl

5.0g

Na2HPO4

2.5g

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

Use: For the cultivation and maintenance of Actinomyces israelii and Propionibacterium propionicus.

BHI/2 Medium

Composition per liter:

Pancreatic digest of gelatin

14.5g

Casein hydrolysate

10.0g

Glucose

8.0g

Brain heart, solids from infusion

6.0g

Peptic digest of animal tissue

6.0g

NaCl

5.0g

Yeast extract

5.0g

Na2HPO4

2.5g

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

Use: For the cultivation and maintenance of Actinomyces georgiae, Actinomyces gerencseriae, Actinomyces naeslundii, Actinomyces odontolyticus, and Actinomyces viscosus.

BHI/3 Medium

Composition per liter:

Pancreatic digest of gelatin

14.5g

Casein hydrolysate

10.0g

Brain heart, solids from infusion

6.0g

Peptic digest of animal tissue

6.0g

Starch

5.0g

NaCl

5.0g

Glucose

3.0g

Na2HPO4

2.5g

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

Use: For the cultivation and maintenance of Actinomyces bovis, Actinomyces gerencseriae, Actinomyces naeslundii, Actinomyces odontolyticus, Actinomyces viscosus, and Streptomyces species.

BHI with Serum and Glucose

See: Brain Heart Infusion with Serum and Glucose

BHIS

See: Brain Heart Infusion, Supplemented

BHIV Agar, 1/10

See: Brain Heart Infusion Agar, 1/10 with Vitamins

BHIY Media

Composition per liter:

Beef heart, infusion from

250.0g

Calf brains, infusion from

200.0g

Yeast extract

20.0g

Agar

15.0g

Proteose peptone

10.0g

Sodium phosphate

2.5g

Dextrose

0.2g

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 Streptococcus bovis and Streptococcus equinus.

Bicarbonate Agar

Composition per 100.0mL:

Soybean-casein digest agar

90.0mL

Sodium bicarbonate solution

10.0mL

pH 7.3 ± 0.2 at 25°C

Soybean-Casein Digest Agar :

Composition per liter:

Agar

15.0g

Pancreatic digest of casein

15.0g

Papaic digest of soybean meal

5.0g

NaCl

5.0g

Preparation of Soybean-Casein Digest Agar: 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.

Sodium Bicarbonate Solution:

Composition per 10.0mL:

NaHCO3

0.7g

Preparation of Sodium Bicarbonate Solution: Add NaHCO3 to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Use freshly prepared solution.

Preparation of Medium: To 90.0mL of cooled, sterile soybean-casein digest agar, aseptically add 10.0mL of sterile sodium bicarbonate solution. Mix thoroughly. Pour into sterile Petri dishes.

Use: For the cultivation of Vibrio species from foods.

Bifidobacterium Medium

Composition per liter:

Glucose

20.0g

Pancreatic digest of casein

20.0g

Yeast extract

10.0g

Peptone

10.0g

Tomato juice

333.0mL

Tween™ 80

2.0mL

pH 6.8 ± 0.2 at 25°C

Preparation of Medium: Combine 333.0mL of tomato juice with 666.0mL of distilled/deionized water. Bring to boiling. Filter through paper. Add remaining components to filtrate. Mix thoroughly. Bring volume to 1.0L with distilled/deionized water. Distribute into tubes or flasks. Autoclave for 30 min at 15 psi pressure–110°C.

Use: For the cultivation of Bifidobacterium infantis.

Bifidobacterium Agar

Composition per liter:

Peptone, special

23.0g

Agar

15.0g

Glucose

5.0g

NaCl

5.0g

Starch, soluble

1.0g

L-Cysteine hydrochloride

0.3g

pH 6.8 ± 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. Distribute into tubes or flasks. Auto-clave for 15 min at 15 psi pressure–121°C. Pour into Petri dishes or leave in tubes.

Use: For the cultivation of Bifidobacterium spp.

Bifidobacterium Broth

Composition per liter:

Glucose

20.0g

Casein enzymatic hydrolysate

20.0g

Tomato juice, solids

16.65g

Peptic digest of animal tissue

10.0g

Yeast extract

10.0g

Tween™ 80

2.0mL

pH 6.8 ± 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. Distribute into tubes or flasks. Auto-clave for 15 min at 15 psi pressure–121°C.

Use: For the cultivation of Bifidobacterium infantis.

Bifidobacterium Medium

Composition per liter:

Tryptic digest of casein

10.0g

Glucose

10.0g

Beef extract

5.0g

Yeast extract

5.0g

K2HPO4

3.0g

Tween™ 80

1.0mL

Sodium ascorbate solution

25.0mL

L-Cysteine·HCl solution

25.0mL

pH 6.8 ± 0.2 at 25°C

Sodium Ascorbate Solution:

Composition per 25.0mL:

Sodium ascorbate

10.0g

Preparation of Sodium Ascorbate Solution: Add sodium ascorbate to distilled/deionized water and bring volume to 25.0mL. Mix thoroughly. Filter sterilize.

L-Cysteine·HCl Solution:

Composition per 25.0mL:

L-Cysteine·HCl

0.5g

Preparation of L-Cysteine·HCl Solution: Add L-cysteine·HCl to distilled/deionized water and bring volume to 25.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except sodium ascorbate solution and L-cysteine·HCl solution, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically add 25.0mL of sterile sodium ascorbate solution and 25.0mL of sterile L-cysteine·HCl solution. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Medium that has not been freshly prepared should be heated in a steamer for 10 min prior to the addition of ascorbate and L-cysteine.

Use: For the cultivation and maintenance of Bifidobacterium adolescentis, Bifidobacterium angulatum, Bifidobacterium animalis, Bifidobacterium asteroides, Bifidobacterium bifidum, Bifidobacterium boum, Bifidobacterium breve, Bifidobacterium catenulatum, Bifidobacterium choerinum, Bifidobacterium coryneforme, Bifidobacterium cuniculi, Bifidobacterium dentium, Bifidobacterium gallicum, Bifidobacterium indicum, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium magnum, Bifidobacterium merycicum, Bifidobacterium minimum, Bifidobacterium pseudocatenulatum, Bifidobacterium pseudo-longum, Bifidobacterium pullorum, Bifidobacterium ruminantium, Bifidobacterium saeculare, Bifidobacterium subtile, Bifidobacterium suis, and Bifidobacterium thermophilum.

Bifidobacterium Medium

Composition per liter:

Special peptone

23.0g

Agar

15.0g

NaCl

5.0g

Glucose

5.0g

Starch, soluble

1.0g

L-Cysteine·HCl

0.3g

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 numerous Bifidobacterium species.

BiGGY Agar

(Bismuth Sulfite Glucose

Glycerin Yeast Extract Agar)

(Nickerson Medium)

Composition per liter:

Agar

16.0g

Glucose

10.0g

Glycine

10.0g

Bismuth ammonium citrate

5.0g

Na2SO3

3.0g

Yeast extract

1.0g

pH 6.8 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Oxoid Unipath and BD Diagnostic Systems.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly and heat with frequent agitation until boiling. Distribute into tubes or flasks. Do not autoclave. Cool to approximately 45°–50°C. If desired, add 2mg/L of neomycin sulfate. Swirl to disperse the insoluble material and pour into sterile Petri dishes.

Use: For the detection, isolation, and presumptive identification of Candida species. Addition of neomycin helps inhibit bacterial species. Candida albicans appears as brown to black colonies with no pigment diffusion and no sheen. Candida tropicalis appears as dark brown colonies with black centers, black pigment diffusion, and a sheen. Candida krusei appears as shiny, wrinkled, brown to black colonies with yellow pigment diffusion. Candida pseudotropicalis appears as flat, shiny red to brown colonies with no pigment diffusion. Candida parakrusei appears as flat, shiny, wrinkled, dark reddish-brown colonies with light reddish-brown peripheries and a yellow fringe. Candida stellatoidea appears as flat dark brown colonies with a light fringe.

Bile Broth Base, HiVeg with Streptokinase

Composition per liter:

Plant peptone

20.0g

NaCl

5.0g

Synthetic detergent No. V

5.0g

Streptokinase solution

1.0mL

pH 7.1 ± 0.2 at 25°C

Source: This medium, without streptokinase solution, is available as a premixed powder from HiMedia.

Streptokinase Solution:

Composition per 1.0mL:

Streptokinase

100,000 units

Streptokinase Solution: Add streptokinase to distilled/deionized water and bring volume to 1.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Heat gently and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 40°C. Aseptically add 1.0mL of streptokinase solution. Mix thoroughly. If desired carbohydrate may also be added to this medium prior to sterilization.

Use: For the culture of blood clots from patients with suspected enteric fever.

Bile Broth Base with Streptokinase

Composition per liter:

Peptone

20.0g

NaCl

5.0g

Synthetic detergent No. V

5.0g

Streptokinase solution

1.0mL

pH 7.1 ± 0.2 at 25°C

Source: This medium, without streptokinase solution, is available as a premixed powder from HiMedia.

Streptokinase Solution:

Composition per 1.0mL:

Streptokinase

100,000 units

Streptokinase Solution: Add streptokinase to distilled/deionized water and bring volume to 1.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Heat gently and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 40°C. Aseptically add 1.0mL of streptokinase solution. Mix thoroughly. If desired carbohydrate may also be added to this medium prior to sterilization.

Use: For the culture of blood clots from patients with suspected enteric fever.

Bile Esculin Agar

Composition per liter:

Oxgall

20.0g

Agar

15.0g

Pancreatic digest of gelatin

5.0g

Beef extract

3.0g

Esculin

1.0g

Ferric citrate

0.5g

Horse serum

50.0mL

pH 6.8 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Oxoid Unipath and BD Diagnostic Systems.

Preparation of Medium: Add components, except horse serum, to distilled/deionized water and bring volume to 950.0L. Mix thoroughly and heat with frequent agitation until boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add 50.0mL of filter sterilized horse serum. Distribute into sterile Petri dishes or test tubes. Cool tubes in a slanted position.

Use: For differentiation between group D streptococci and nongroup D streptococci. To differentiate members of the Enterobacteriaceae, particularly Klebsiella, Enterobacter, and Serratia, from other enteric bacteria. To differentiate Listeria monocytogenes. Bile tolerance and esculin hydrolysis (seen as a dark brown to black complex) are presumptive for enterococci (group D streptococci).

Bile Esculin Agar

Composition per liter:

Esculin

1.0g

Bile esculin agar base

1.0L

pH 6.6 ± 0.2 at 25°C

Bile Esculin Agar Base:

Composition per liter:

Oxgall

40.0g

Agar

15.0g

Peptone

5.0g

Beef extract

3.0g

Ferric citrate

0.5g

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

Preparation of Bile Esculin Agar Base: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

Preparation of Medium: Add desired amount of esculin—typically 1.0g—to bile esculin agar base. Mix thoroughly and heat with frequent agitation until boiling. Autoclave for 15 min at 15 psi pressure– 121°C. Cool to 45°–50°C. Distribute into sterile Petri dishes or test tubes. Cool tubes in a slanted position.

Use: For the isolation and presumptive identification of group D streptococci.

Bile Esculin Agar

(BAM M18)

Composition per liter:

Oxgall

40.0g

Agar

15.0g

Pancreatic digest of gelatin

5.0g

Beef extract

3.0g

Esculin

1.0g

Ferric citrate

0.5g

pH 6.6 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly and heat with frequent agitation until boiling. Autoclave for 15 min at 15 psi pressure–121°C. Distribute into sterile Petri dishes or test tubes. Cool tubes in a slanted position.

Use: For differentiation between group D streptococci and nongroup D streptococci. To differentiate members of the Enterobacteriaceae, particularly Klebsiella, Enterobacter, and Serratia, from other enteric bacteria. To differentiate Listeria monocytogenes. Bile tolerance and esculin hydrolysis (seen as a dark brown to black complex) are presumptive for enterococci (group D streptococci).

Bile Esculin Agar

Composition per liter:

Bile salts

40.0g

Agar

15.0g

Pancreatic digest of animal tissue

5.0g

Beef extract

3.0g

Esculin

1.0g

Ferric citrate

0.5g

pH 6.6 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly and heat with frequent agitation until boiling. Autoclave for 15 min at 15 psi pressure–121°C. Distribute into sterile Petri dishes or test tubes.

Use: For the isolation and identification of Yersinia enterocolitica.

Bile Esculin Agar, HiVeg

Composition per liter:

Plant peptone

25.0g

Agar

15.0g

Plant hydrolysate

15.0g

Plant extract

6.0g

Synthetic detergent No. II

2.0g

Esculin

1.0g

Ferric citrate

0.5g

pH 6.6 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Hi-Media.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly and heat with frequent agitation until boiling. Autoclave for 15 min at 15 psi pressure–121°C. Distribute into sterile Petri dishes or test tubes. Cool tubes in a slanted position.

Use: For the isolation and presumptive identification of group D streptococci.

Bile Esculin Agar with Kanamycin

Composition per liter:

Oxgall

20.0g

Agar

15.0g

Beef extract

3.0g

Esculin

1.0g

Ferric citrate

0.5g

Hemin

10.0mg

Vitamin K1

10.0mg

Horse serum

50.0mL

Kanamycin solution

10.0mL

pH 7.1 ± 0.2 at 25°C

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

Kanamycin Solution:

Composition per 10.0mL:

Kanamycin

1.0g

Preparation of Kanamycin Solution: Add kanamycin to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly and heat with frequent agitation until boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add 50.0mL of 5% filter-sterilized horse serum and 10.0mL of sterile kanamycin solution. Distribute into test tubes or flasks. Cool tubes in a slanted position.

Use: For the selective isolation and/or presumptive identification of bacteria of the Bacteroides fragilis group from specimens containing mixed flora. Examine colonies with a long-wavelength UV light. Pigmented colonies of the Bacteroides group will fluoresce red-orange. Growth on this medium with blackening of the medium is presumptive for Bacteroides fragilis.

Bile Esculin Azide Agar

Composition per liter:

Pancreatic digest of casein

17.0g

Agar

15.0g

Oxgall

10.0g

NaCl

5.0g

Yeast extract

5.0g

Proteose peptone No. 3

3.0g

Esculin

1.0g

Ferric ammonium citrate

0.5g

NaN3

0.15g

pH 7.1 ± 0.2 at 25°C

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

Caution: Sodium azide is toxic. Azides also react with metals and disposal must be highly diluted.

Preparation: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly and heat with frequent agitation until boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Pour into sterile Petri dishes or leave in tubes. Cool tubes in a slanted position.

Use: For the isolation and presumptive identification of group D streptococci.

Bile Esculin Azide HiVeg Agar

Composition per liter:

Plant hydrolysate

20.0g

Agar

15.0g

Plant extract

5.0g

Plant peptone No. 3

5.0g

NaCl

5.0g

Synthetic detergent No. II

5.0g

Esculin

1.0g

Ferric ammonium citrate

0.5g

NaN3

0.15g

pH 7.1 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Hi-Media.

Caution: Sodium azide is toxic. Azides also react with metals and disposal must be highly diluted.

Preparation: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly and heat with frequent agitation until boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Pour into sterile Petri dishes or leave in tubes. Cool tubes in a slanted position.

Use: For the isolation and presumptive identification of group D streptococci.

Bile Esculin HiVeg Agar Base with Esculin

Composition per liter:

Plant peptone

22.0g

Agar

15.0g

Plant hydrolysate

15.0g

Plant extract

6.0g

Synthetic detergent No. II

5.0g

Ferric citrate

0.5g

Esculin solution

4.0mL

pH 6.8 ± 0.2 at 25°C

Source: This medium, without esculin, is available as a premixed powder from HiMedia.

Esculin Solution:

Composition per 4.0mL:

Esculin

1.0g

Esculin Solution: Add esculin to distilled/deionized water and bring volume to 4.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except esculin solution, 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. Aseptically add 4.0mL of sterile escu-lin solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile test tubes.

Use: For the isolation and presumptive identification of group D streptococci.

Bile Esculin HiVeg Agar with Kanamycin

Composition per liter:

Plant peptone no. 2

17.0g

Agar

15.0g

Plant extract

6.0g

Synthetic detergent

5.0g

Esculin

1.0g

Ferric citrate

0.5g

Kanamycin

0.1g

Fe4(P2O7)3·H2O

0.01g

Vitamin K1

0.01g

pH 7.1 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Hi-Media.

Preparation: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly and heat with frequent agitation until boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Pour into sterile Petri dishes or leave in tubes. Cool tubes in a slanted position.

Use: For the selective isolation and/or presumptive identification of bacteria of the Bacteroides fragilis group from specimens containing mixed flora. Examine colonies with a long-wavelength UV light. Pigmented colonies of the Bacteroides group will fluoresce red-orange. Growth on this medium with blackening of the medium is presumptive for Bacteroides fragilis.

Bile Oxalate Sorbose Broth

(BOS Broth)

Composition per liter:

Na2HPO4

9.14g

Sodium oxalate

5.0g

Bile salts

2.0g

NaCl

1.0g

CaCl2·2H2O

0.01g

MgSO4·7H2O

0.01g

Asparagine solution

100.0mL

Methionine solution

100.0mL

Sorbose solution

100.0mL

Yeast extract solution

10.0mL

Sodium pyruvate solution

10.0mL

Metanil Yellow solution

10.0mL

Sodium nitrofurantoin solution

10.0mL

Irgasan® solution

1.0mL

pH 7.6 ± 0.2 at 25°C

Asparagine Solution:

Composition per 100.0mL:

Asparagine

1.0g

Preparation of Asparagine Solution: Add asparagine to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

Methionine Solution:

Composition per 100.0mL:

Methionine

1.0g

Preparation of Methionine Solution: Add methionine to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

Sorbose Solution:

Composition per 100.0mL:

Sorbose

10.0g

Preparation of Sorbose Solution: Add sorbose to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

Yeast Extract Solution:

Composition per 10.0mL:

Yeast extract

0.025g

Preparation of Yeast Extract Solution: Add yeast extract to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Sodium Pyruvate Solution:

Composition per 10.0mL:

Sodium pyruvate

0.05g

Preparation of Sodium Pyruvate Solution: Add sodium pyruvate to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Metanil Yellow Solution:

Composition per 10.0mL:

Metanil Yellow

0.025g

Preparation of Metanil Yellow Solution: Add Metanil Yellow to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Sodium Nitrofurantoin Solution:

Composition per 10.0mL:

Sodium nitrofurantoin

0.01g

Preparation of Sodium Nitrofurantoin Solution: Add sodium nitrofurantoin to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Irgasan® Solution:

Composition per 10.0mL:

Irgasan

0.04g

Ethanol (95% solution)

10.0mL

Preparation of Irgasan Solution: Add Irgasan to 10.0mL of ethanol. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except asparagine solution, methionine solution, sorbose solution, yeast extract solution, sodium pyruvate solution, Metanil Yellow solution, sodium nitrofurantoin solution, and Irgasan solution, to distilled/deionized water and bring volume to 659.0mL. 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 asparagine solution, 100.0mL of sterile methionine solution, 100.0mL of sterile sorbose solution, 10.0mL of sterile yeast extract solution, 10.0mL of sterile sodium pyruvate solution, 10.0mL of sterile Metanil Yellow solution, 10.0mL of sterile sodium nitrofurantoin solution, and 1.0mL of sterile Irgasan solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the isolation and cultivation of Yersinia enterocolitica from foods.

Bile Peptone Transport Medium

Composition per liter:

Casein enzymatic hydrolysate

10.0g

NaCl

10.0g

Sodium taurocholate

5.0g

pH 8.5 ± 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. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the transport and preservation of Vibrio cholerae.

Bile Salt Agar with Streptokinase

Composition per liter:

Agar

18.0g

Peptone

10.0g

Meat extract

10.0g

NaCl

5.0g

Sodium taurocholate

5.0g

Streptokinase solution

1.0mL

pH 8.2 ± 0.2 at 25°C

Source: This medium, without streptokinase solution, is available as a premixed powder from HiMedia.

Streptokinase Solution:

Composition per 1.0mL:

Streptokinase

100,000 U

Streptokinase Solution: Add streptokinase to distilled/deionized water and bring volume to 1.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Heat gently and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 40°C. Aseptically add 1.0mL of streptokinase solution. Mix thoroughly. If desired carbohydrate may also be added to this medium prior to sterilization.

Use: For the isolation and cultivation of bile tolerant enteric bacilli.

Bile Salts Brilliant Green Starch Agar

(BBGS Agar)

Composition per liter:

Agar

15.0g

Soluble starch

10.0g

Proteose peptone

10.0g

Beef extract

5.0g

Bile salts

5.0g

Brilliant Green (0.05% solution)

1.0mL

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

Use: For the isolation and cultivation of Aeromonas hydrophila from foods.

Bile Salts Gelatin Agar

Composition per 100.0mL:

Gelatin

3.0g

Agar

1.5g

Pancreatic digest of casein

1.0g

NaCl

1.0g

Sodium taurocholate

0.5g

Na2CO3

0.1g

Water

100.0mL

pH 8.5 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation of Vibrio cholerae.

BIN Medium

Composition per liter:

Beef heart, infusion from

250.0g

Calf brains, infusion from

200.0g

Agar

15.0g

Proteose peptone

10.0g

NaCl

5.0g

Na2HPO4

2.5g

Glucose

2.0g

Irgasan solution

4.0mL

Crystal Violet solution

1.0mL

Sodium cholate solution

1.0mL

Sodium deoxycholate solution

1.0mL

Nystatin solution

1.0mL

pH 7.4 ± 0.2 at 25°C

Sodium Cholate Solution:

Composition per 100.0mL:

Sodium cholate

5.0g

Preparation of Sodium Cholate Solution: Add sodium cholate to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Gently heat while stirring and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.

Sodium Deoxycholate Solution:

Composition per 100.0mL:

Sodium deoxycholate

5.0g

Preparation of Sodium Deoxycholate Solution: Add sodium deoxycholate to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Gently heat while stirring and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool 25°C.

Irgasan Solution:

Composition per 50.0mL:

Irgasan DP300

10.0mg

Ethanol, 90%

50.0mL

Preparation of Irgasan Solution: Add irgasan to 90% ethanol and bring volume to 50.0mL. Mix thoroughly.

Crystal Violet Solution:

Composition per 10.0mL:

Crystal Violet

10.0mg

Preparation of Crystal Violet Solution: Add Crystal Violet to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Gently heat while stirring and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.

Nystatin Solution:

Composition per 10.0mL:

Nystatin

2.5g

Preparation of Nystatin Solution: Add nystatin to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except irgasan solution, Crystal Violet solution, sodium cholate solution, sodium deoxycholate solution, and nystatin solution, to distilled/deionized water and bring volume to 992.0mL. Mix thoroughly. Gently heat while stirring and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 85°C. Aseptially add 4.0mL irgasam solution. Mix thoroughly to volatilize the ethanol. Cool to 50°C. Aseptially add 1.0mL each of Crystal Violet solution, sodium cholate solution, sodium deoxycholate solution, and nystatin solution. Mix thoroughly. Pour into sterile Petri dishes.

Use: For the efficient detection of Yersinia pestis from clinical and other specimens. The formulation of this medium is based on brain heart infusion agar, to which the selective agents irgasan, cholate salts, Crystal Violet, and nystatin are introduced to enhance efficiency of recovery of Y. pestis.

Biosynth Chromogenic Medium

for Listeria monocytogenes

(BCM for Listeria monocytogenes)

(BAM M17a)

Composition per liter:

Proprietary

Source: This medium is available from Biosynth International, Inc.

Use: To differentiate Listeria monocytogenes and L. ivanovii from other Listeria spp. Supplements render the medium selective. Differential activity for all Listeria species is based upon a chromogenic substrate included in the medium. This is a complete test system with a fluorogenic selective enrichment broth and a chromogenic plating medium both detecting the virulence factor phosphatidylinositol specific phospholipase C (PI-PLC). The medium contains a substrate for phosphatidylinositol-specific phospholipase C (PlcA) enzymes. The selective enrichment broth is fluorogenic. The plating medium for rapid detection and enumeration of pathogenic Listeria combines cleavage of the chromogenic PI-PLC substrate with the additional production of a white precipitate surrounding the target colonies.

Biotin Assay Medium

Composition per liter:

Glucose

40.0g

Sodium acetate

20.0g

Vitamin assay casamino acids

12.0g

K2HPO4

1.0g

KH2PO4

1.0g

MgSO4·7H2O

0.4g

DL-Tryptophane

0.2g

L-Cystine

0.2g

Adenine sulfate

0.02g

FeSO4

0.02g

Guanine·HCl

0.02g

MgSO4·7H2O

0.02g

NaCl

0.02g

Uracil

0.02g

Calcium pantothenate

2.0mg

Niacin

2.0mg

Pyridoxine·HCl

2.0mg

Riboflavin

2.0mg

Thiamine·HCl

2.0mg

p-Aminobenzoic acid

0.2mg

pH 6.7 ± 0.2 at 25°C

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

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 tubes in 5.0mL volumes. Add standard solution or test solutions to each tube. Adjust the volume of each tube to 10.0mL with distilled/deionized water. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For use in the microbiological assay of biotin using Lactobacillus plantarum as the test microorganism.

Biphasic Medium for Neisseria

Composition per liter:

Glucose starch agar

1.0L

Glucose starch broth

1.0L

pH 7.3 ± 0.2 at 25°C

Glucose Starch Agar:

Composition per liter:

Agar

20.0g

Gelatin

20.0g

Proteose peptone No. 3

15.0g

Soluble starch

10.0g

NaCl

5.0g

Glucose

2.0g

Na2HPO4

3.0g

Preparation of Glucose Starch Agar: 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.

Glucose Starch Broth:

Composition per liter:

Gelatin

20.0g

Proteose peptone No. 3

15.0g

Soluble starch

10.0g

NaCl

5.0g

Glucose

2.0g

Na2HPO4

3.0g

Preparation of Glucose Starch Broth: 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 25°C.

Preparation of Medium: Aseptically distibute glucose starch agar into flasks in 100–125mL volumes. Allow agar to solidify. Overlay agar with 25.0mL of sterile glucose starch broth.

Use: For selective isolation and cultivation of Neisseria species.

Biphenyl Agar

(DSMZ Medium 457d)

Composition per liter:

Agar

15.0g

Na2HPO4

2.44g

KH2PO4

1.52g

(NH4)2SO4

0.5g

Biphenyl

0.25g

MgSO4·7H2O

0.2g

CaCl2·2H2O

0.05g

Trace elements solution SL-4

10.0mL

pH 6.9 ± 0.2 at 25°C

Trace Elements Solution SL-4:

Composition per liter:

EDTA

0.5g

FeSO4·7H2O

0.2g

Trace elements solution SL-6

100.0mL

Trace Elements Solution SL-6:

Composition per liter:

H3BO3

0.3g

CoCl2·6H2O

0.2g

ZnSO4·7H2O

0.1g

MnCl2·4H2O

0.03g

Na2MoO4·H2O

0.03g

NiCl2·6H2O

0.02g

CuCl·2H2O

0.01g

Preparation of Trace Elements Solution SL-6: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 3.4.

Preparation of Trace Elements Solution SL-4: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

Biphenyl Solution:

Composition per liter:

Biphenyl

10.0g

Preparation of Biphenyl Solution: Add biphenyl to 1.0L ethanol. Mix thoroughly. Filter sterilize using a cellulose filter membrane.

Preparation of Medium: Add components, except biphenyl solution, to 1.0L distilled/deionized water. Adjust pH to 6.9. Heat and gently bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Add an aliquot of the biphenyl solution to the lid of a sterile Petri dish so that the final concentration will be approximately 0.25g/L biphenyl, and let the ethanol evaporate so that the crystals of biphenyl coat the lid of the Petri dish. Aseptically add sterile agar medium to the crystal-layered Petri dish.

Use: For the cultivation of biphenyl-utilizing bacteria.

Bird Seed Agar

( Guizotia abyssinica Creatinine Agar)

(Niger Seed Agar)/(Staib Agar)

Composition per liter:

Agar

15.0g

Glucose

15.0g

Creatinine

5.0g

KH2PO4

3.0g

Biphenyl

1.0g

Chloramphenicol

0.5g

Guizotia abyssinica seed (niger seed) extract

1000.0mL

pH 6.7 ± 0.2 at 25°C

Preparation of Medium: Prepare seed extract by grinding 50.0g of Guizotia abyssinica seed in 1.0L of distilled/deionized water. Boil for 30 min. Filter through cheesecloth and filter paper. Add remaining components to seed filtrate. Mix thoroughly and heat with frequent agitation until boiling. Distribute into flasks or tubes. Autoclave for 25 min at 15 psi pressure–110°C.

Use: For the selective isolation and differentiation of Cryptococcus neoformans from other Cryptococcus species and other yeasts.

Bismuth Sulfite Agar

Composition per liter:

Agar

20.0g

Bi2(SO3)3

8.0g

Pancreatic digest of casein

5.0g

Peptic digest of animal tissue

5.0g

Beef extract

5.0g

Glucose

5.0g

Na2HPO4

4.0g

FeSO4·7H2O

0.3g

pH 7.5 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Oxoid Unipath and BD Diagnostic Systems.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly and heat with frequent agitation until boiling. Boil for 1 min. Do not autoclave. Cool to 45°– 50°C. Pour into sterile Petri dishes while gently shaking flask to disperse precipitate. Use plates the same day as prepared.

Use: For the selective isolation and identification of Salmonella typhi and other enteric bacilli. Salmonella typhi appears as flat, black, “rabbit-eye” colonies surrounded by a zone of black with a metallic sheen.

Bismuth Sulfite Agar

Composition per liter:

Agar

20.0g

Peptic digest of animal tissue

10.0g

Bismuth sulfite indicator

8.0g

Glucose

5.0g

Beef extract

5.0g

Na2HPO4

4.0g

FeSO4

0.3g

Brilliant Green

0.025g

pH 7.7 ± 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 and heat with frequent agitation until boiling. Boil for 1 min. Do not autoclave. Cool to 45°– 50°C. Mix thoroughly. The sensitivity of the medium depends largely upon uniform dispersion of precipitated bismuth sulfite in the final gel which should be dispersed before pouring the plates. Pour into sterile Petri dishes while gently shaking flask to disperse precipitate. Use plates the same day as prepared.

Use: For the selective isolation and identification of Salmonella typhi and other enteric bacilli. Salmonella typhi appears as flat, black, “rabbit-eye” colonies surrounded by a zone of black with a metallic sheen.

Bismuth Sulfite Agar, HiVeg

Composition per liter:

Agar

20.0g

Plant peptone

10.0g

Bismuth sulfite indicator

8.0g

Glucose

5.0g

Plant extract

5.0g

Na2HPO4

4.0g

FeSO4

0.3g

Brilliant Green

0.025g

pH 7.7 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly and heat with frequent agitation until boiling. Boil for 1 min. Do not autoclave. Cool to 45°– 50°C. Mix thoroughly. The sensitivity of the medium depends largely upon uniform dispersion of precipitated bismuth sulfite in the final gel which should be dispersed before pouring the plates. Pour into sterile Petri dishes while gently shaking flask to disperse precipitate. Use plates the same day as prepared.

Use: For the selective isolation and identification of Salmonella typhi and other enteric bacilli. Salmonella typhi appears as flat, black, “rabbit-eye” colonies surrounded by a zone of black with a metallic sheen.

Bismuth Sulfite Agar, Modified

Composition per liter:

Agar

12.7g

Bismuth sulfite indicator

8.0g

Glucose

5.0g

Beef extract

5.0g

Peptic digest of animal tissue

5.0g

Na2HPO4

4.0g

FeSO4

0.3g

Brilliant Green

0.016

pH 7.5 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Hi-Media.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly and heat with frequent agitation until boiling. Boil for 1 min. Do not autoclave. Cool to 45°– 50°C. Mix thoroughly. The sensitivity of the medium depends largely upon uniform dispersion of precipitated bismuth sulfite in the final gel which should be dispersed before pouring the plates. Pour into sterile Petri dishes while gently shaking flask to disperse precipitate. Use plates the same day as prepared.

Use: For the selective isolation and identification of Salmonella typhi and other enteric bacilli. Salmonella typhi appears as flat, black, “rabbit-eye” colonies surrounded by a zone of black with a metallic sheen.

Bismuth Sulfite Agar, Modified, HiVeg

Composition per liter:

Agar

12.7g

Bismuth sulfite indicator

8.0g

Glucose

5.0g

Plant extract

5.0g

Plant peptone

5.0g

Na2HPO4

4.0g

FeSO4

0.3g

Brilliant Green

0.016

pH 7.5 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Hi-Media.

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

agitation until boiling. Boil for 1 min. Do not autoclave. Cool to 45°– 50°C. Mix thoroughly. The sensitivity of the medium depends largely upon uniform dispersion of precipitated bismuth sulfite in the final gel which should be dispersed before pouring the plates. Pour into sterile Petri dishes while gently shaking flask to disperse precipitate. Use plates the same day as prepared.

Use: For the selective isolation and identification of Salmonella typhi and other enteric bacilli. Salmonella typhi appears as flat, black, “rabbit-eye” colonies surrounded by a zone of black with a metallic sheen.

Bismuth Sulfite Agar Wilson and Blair

(BAM 19)

Composition per liter:

Agar

20.0g

Pancreatic digest of casein

10.0g

Bi2(SO3)3

8.0g

Beef extract

5.0g

Glucose

5.0g

Na2HPO4

4.0g

FeSO4·7H2O

0.3g

Brilliant Green

0.025g

pH 7.7 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly and heat with frequent agitation until boiling. Boil for 1 min. Do not autoclave. Cool to 45°– 50°C. Pour into sterile Petri dishes while gently shaking flask to disperse precipitate. Let plates dry for about 2h with lids partially removed. Use plates the within one day of preparation; medium loses selectivity after 48h.

Use: For the selective isolation and identification of Salmonella typhi and other enteric bacilli. Salmonella typhi appears as flat, black, “rabbit-eye” colonies surrounded by a zone of black with a metallic sheen.

Bismuth Sulfite Broth

(m-Bismuth Sulfite Broth)

Composition per liter:

Bi2(SO3)3

16.0g

Pancreatic digest of casein

10.0g

Peptic digest of animal tissue

10.0g

Beef extract

10.0g

Glucose

10.0g

Na2HPO4

8.0g

FeSO4·7H2O

0.6g

pH 7.7 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly and heat with frequent agitation until boiling. Boil for 1 min. Do not autoclave. Cool to 45°– 50°C. Mix to disperse the precipitate and aseptically distribute into sterile tubes or flasks. Use 2.0–2.2mL of medium for each membrane filter.

Use: For the selective isolation of Salmonella typhi and other enteric bacilli and for the detection of Salmonella by the membrane filter method.

Bismuth Sulfite Glucose

Glycerin Yeast Extract Agar

See: BiGGY Agar

BL Agar

(Glucose Blood Liver Agar)

Composition per liter:

Agar

15.0g

Glucose

10.0g

Proteose peptone No. 3

10.0g

Pancreatic digest of casein

5.0g

Yeast extract

5.0g

Meat extract

3.0g

Phytone™

3.0g

Tween™ 80

1.0g

Soluble starch

0.5g

Liver extract

150.0mL

Horse blood

50.0mL

L-Cysteine·HCl solution

10.0mL

Solution A

10.0mL

Solution B

5.0mL

pH 7.2 ± 0.2 at 25°C

Liver Extract:

Composition per 170.0mL:

Liver powder

10.0g

Preparation of Liver Extract: Add 10.0g of liver powder to 170mL of distilled/deionized water. Gently heat to 60°C. Maintain at 50°–60°C for 1 hr. Gently bring to boiling. Boil for 5 min. Adjust pH to 7.2. Filter through Whatman #2 filter paper.

L-Cysteine·HCl Solution:

Composition per 10.0mL:

L-Cysteine·HCl

0.5g

Preparation of L-Cysteine·HCl Solution: Dissolve 0.5g of L-cysteine·HCl in distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Warm to 50°C.

Solution A:

Composition per 100.0mL:

K2HPO4

10.0g

KH2PO4

10.0g

Preparation of Solution A: Add components to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C.

Solution B:

Composition per 100.0mL:

MgSO4·7H2O

4.0g

NaCl

0.2g

FeSO4·7H2O

0.2g

MnSO4·H2O

0.2g

Preparation of Solution B: Add components to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C.

Preparation of Medium: Add components, except liver extract, horse blood, L-cysteine·HCl solution, solution A, and solution B, to distilled/deionized water and bring volume to 775.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Aseptically add 150.0mL of sterile liver extract, 50.0mL of sterile horse blood, 10.0mL of sterile L-cysteine·HCl solution, 10.0 mL of sterile solution A, and 5.0mL of sterile solution B. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation and maintenance of Atopobium minutum, Bacteroides distasonis, Bacteroides ovatus, Bacteroides thetaiotaomicron, Bacteroides uniformis, Bacteroides vulgatus, numerous Bifido-bacterium species, Campylobacter divergens, Carnobacterium piscicola, numerous Clostridium species, numerous Lactobacillus species, Lactococcus lactis, Leuconostoc lactis, Leuconostoc mesenteroides, and Propionibacterium thoenii.

Blaser’s Agar

See: Campylobacter Selective Medium, Blaser-Wang

Blaser’s Campylobacter Agar

See: Campylobacter Agar, Blaser’s

Blaser-Wang Campylobacter Medium

See : Blaser-Wang

Blaser-Wang Campylobacter Medium

See : Campylobacter Selective Medium, Blaser-Wang

Blastobacter denitrificans Agar

(LMG Medium 157)

Composition per liter:

Agar

15.0g

Tryptone

2.0g

Lab Lemco beef extract

0.5g

Yeast extract

0.5g

Sodium acetate

0.2g

Glucose solution

10.0mL

pH 7.3 ± 0.2 at 25°C

Glucose Solution:

Composition per 10.0mL:

Glucose

2.5g

Preparation of Glucose Solution: Add glucose to 10.0mL of distilled/deionized water. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except glucose 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 45°–50°C. Aseptically add 10.0mL glucose solution. Mix thoroughly. Aseptically pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation of Blastobacter denitrificans.

Blastobacter denitrificans Agar

Composition per liter:

Agar

15.0g

Pancreatic digest of casein

2.0g

Beef extract

0.5g

Yeast extract

0.5g

Sodium acetate

0.2g

Glucose solution

10.0mL

pH 7.2 ± 0.2 at 25°C

Glucose Solution:

Composition per 10.0mL:

Glucose

2.5g

Preparation of Glucose Solution: Add glucose to distilled/deionized water and bring volume to 10.0L. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except glucose solution, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Aseptically add 10.0mL of sterile glucose solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation of Blastobacter denitrificans.

Blastobacter Enrichment Medium

Composition per liter:

Agar

18.0g

Peptone

0.5g

MgSO4·7H2O

0.13g

KH2PO4·3H2O

0.13g

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components to tap 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 enrichment and cultivation of Blastobacter species.

Blastobacter Medium

Composition per liter:

Agar

15.0g

Peptone

10.0g

Yeast extract

10.0g

NaCl

5.0g

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly and heat with frequent agitation until boiling. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the cultivation and maintenance of Blastobacter natatorius and other Blastobacter species.

Blastococcus aggregatus Medium

Composition per 1001.0mL:

Tryptone

2.0g

Yeast extract

2.0g

Tris(hydroxymethyl)amino methane·HCl buffer

1.0g

KNO3

0.5g

Sodium glycerophosphate

0.1g

Artificial seawater

1.0L

Trace elements solution

1.0mL

pH 7.0 ± 0.2 at 25°C

Artificial Seawater:

Composition per liter:

Commercially available marine aquarium salts mixture

variable

Preparation of Artificial Seawater: Add commercially available marine aquarium salts mixture. Prepare according to manufacturer’s recommendations. Mix thoroughly.

Trace Elements Solution:

Composition per liter:

H3BO3

2.85g

MnCl2·4H2O

1.8g

Sodium tartrate

1.77g

FeSO4

1.36g

CoCl2·6H2O

40.4mg

CuCl2·2H2O

26.9mg

Na2MoO4·2H2O

25.2mg

ZnCl2

20.8mg

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

Preparation of Medium: Combine components. 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 Blastococcus aggregatus.

Blastocystis Egg Medium

Composition per 1300.0mL:

Homogenized whole egg

783.0mL

Stone’s modification of Locke’s solution

217.0mL

Horse serum, heat inactivated

300.0mL

Homogenized Whole Egg:

Composition per liter:

Whole eggs

18–24

Preparation of Homogenized Whole Egg: Use fresh fertile eggs, less than 1 week old. Scrub the shells with soap. Let stand in a soap solution for 30 min. Rinse in running water. Soak eggs in 70% ethanol for 15 min. Break the eggs into a sterile container. Homogenize by shaking. Filter through four layers of sterile cheesecloth into a sterile graduated cylinder. Measure out 1.0L.

Stone’s Modification of Locke’s Solution:

Composition per liter:

NaCl

8.0g

Na2HPO4

2.0g

NaHCO3

0.4g

KH2PO4

0.3g

CaCl2

0.2g

KCl

0.2g

MgCl2·6H2O

0.01g

Preparation of Stone’s Modification of Locke’s Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

Preparation of Medium: Distribute homogenized whole egg in 4.0mL volumes into 16 × 125mm screw-capped test tubes. Place tubes in a slanted position. Inspissate at 80°C (moist heat) for 10 min. Allow to cool. Add 4.5mL of Stone’s modification of Locke’s solution to the surface of the solidified egg in each tube. Close tubes with a rubber stopper. Place tubes in a press. Autoclave for 15 min at 15 psi pressure– 121°C. Cool to room temperature. Aseptically replace rubber stoppers with sterile screw caps. Prior to use, aseptically add 1.5mL of heat-inactivated sterile horse serum to each tube.

Use: For the cultivation of Anophryoides species, Blastocystis hominis, other Blastocystis species, Endolimax nana, and Metanophrys species.

BLE HiVeg Broth Base

with Listeria Selective Supplement

(Buffered Listeria Enrichment HiVeg Broth Base

with Listeria Selective Supplement)

Composition per liter:

Plant hydrolysate

17.0g

Na2HPO4, anhydrous

9.6g

Yeast extract

6.0g

NaCl

5.0g

Papaic digest of soybean meal

3.0g

KH2PO4

2.5g

Glucose

2.5g

Sodium pyruvate

1.0g

Listeria selective supplement

5.0mL

pH 7.3 ± 0.2 at 25°C

Listeria Selective Supplement:

Composition per 5.0mL:

Cycloheximide

50.0mg

Nalidixic acid

40.0mg

Acriflavin hydrochloride

15.0mg

Preparation of Listeria Selective Supplement: Add components to distilled/deionized water and bring volume to 5.0mL. Mix thoroughly. Filter sterilize.

Caution: Cycloheximide is toxic. Avoid skin contact or aerosol formation and inhalation.

Source: This medium, without Listeria selective supplement, is available as a premixed powder from HiMedia.

Preparation of Medium: Add components, except Listeria selective supplement, 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 45°–50°C. Aseptically add 50.0mL of sterile Listeria selective supplement. Mix thoroughly.

Use: For the enrichment and isolation of Listeria monocytogenes.

Blood Agar

Composition per liter:

Agar

15.0g

Pancreatic digest of casein

15.0g

Papaic digest of soybean meal

5.0g

NaCl

5.0g

Sheep blood, defibrinated

50.0mL

pH 7.6 ± 0.2 at 25°C

Preparation of Medium: Add components, except sheep blood, to distilled/deionized water and bring volume to 950.0mL. 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 50.0mL of sterile sheep blood. Mix thoroughly. Pour into sterile Petri dishes in 20.0mL volumes.

Use: For the cultivation of fastidious microorganisms.

Blood Agar Base

Composition per liter:

Agar

15.0g

Beef extract

10.0g

Peptone

10.0g

NaCl

5.0g

Sheep blood, defibrinated

50.0mL

pH 7.3 ± 0.2 at 25°C

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

Preparation of Medium: Add components, except sheep blood, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Heat with frequent agitation and boil for 1 min to completely dissolve. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°– 50°C. Aseptically add 50.0mL of sterile, defibrinated sheep blood. Mix thoroughly and pour into sterile Petri dishes.

Use: For the isolation, cultivation, and detection of hemolytic activity of streptococci and other fastidious microorganisms.

Blood Agar Base

(ATCC Medium 368)

Composition per liter:

Beef heart, infusion from

500.0g

Agar

15.0g

Tryptose

10.0g

NaCl

5.0g

pH 6.8 ± 0.2 at 25°C

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

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Heat with frequent agitation and boil for 1 min to completely dissolve. Autoclave for 15 min at 15 psi pressure–121°C. Cool the basal medium to 45°–50°C. Aseptically add sterile, defibrinated blood to a final concentration of 5%. Mix thoroughly and pour into sterile Petri dishes.

Use: For the isolation, cultivation, and detection of hemolytic activity of staphylococci, streptococci, and other fastidious microorganisms.

Blood Agar Base

(BAM M20a)

Composition per liter:

Beef heart, infusion from

500.0g

Agar

15.0g

Tryptose

10.0g

NaCl

5.0g

Sheep blood, defibrinated

50.0mL

pH 6.8 ± 0.2 at 25°C

Preparation of Medium: Add components, except sheep blood, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Heat with frequent agitation and boil for 1 min to completely dissolve. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°– 50°C. Aseptically add 50.0mL of sterile, defibrinated sheep blood. Mix thoroughly and pour into sterile Petri dishes.

Use: For the isolation, cultivation, and detection of hemolytic activity of staphylococci, streptococci, and other fastidious microorganisms.

Blood Agar Base

(Infusion Agar)

Composition per liter:

Agar

15.0g

Pancreatic digest of casein

13.0g

NaCl

5.0g

Yeast extract

5.0g

Heart muscle, solids from infusion

2.0g

Sheep blood, defibrinated

50.0mL

pH 7.3 ± 0.2 at 25°C

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

Preparation of Medium: Add components, except sheep blood, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Heat with frequent agitation and boil for 1 min to completely dis-solve. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°– 50°C. Aseptically add 50.0mL of sterile, defibrinated sheep blood. Mix thoroughly and pour into sterile Petri dishes.

Use: For the isolation, cultivation, and detection of hemolytic activity of streptococci and other fastidious microorganisms.

Blood Agar Base

(Infusion Agar)

(FDA Medium M21)

Composition per liter:

Heart muscle, infusion from

375.0g

Agar

15.0g

Thiotone

10.0g

NaCl

5.0g

pH 7.3 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 20 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.

Use: For the cultivation of a variety of microorganisms. For the preparation of blood agar by the addition of sterile blood.

Blood Agar Base with Blood

Composition per liter:

Agar

15.0g

Beef extract

10.0g

Tryptose

10.0g

NaCl

5.0g

Sheep blood, defibrinated

50.0mL

pH 7.3 ± 0.2 at 25°C

Source: This medium without blood is available as a premixed powder from HiMedia.

Preparation of Medium: Add components, except sheep blood, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Heat with frequent agitation and boil for 1 min to completely dissolve. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°– 50°C. Aseptically add 50.0mL of sterile, defibrinated sheep blood. Mix thoroughly and pour into sterile Petri dishes.

Use: For the isolation, cultivation, and detection of hemolytic activity of streptococci and other fastidious microorganisms.

Blood Agar Base, HiVeg with Blood

Composition per liter:

Agar

15.0g

Plant hydrolysate No. 1

10.0g

Plant infusion

10.0g

NaCl

5.0g

Sheep blood, defibrinated

50.0mL

pH 7.3 ± 0.2 at 25°C

Source: This medium without blood is available as a premixed powder from HiMedia.

Preparation of Medium: Add components, except sheep blood, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Heat with frequent agitation and boil for 1 min to completely dissolve. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°– 50°C. Aseptically add 50.0mL of sterile, defibrinated sheep blood. Mix thoroughly and pour into sterile Petri dishes.

Use: For the isolation, cultivation, and detection of hemolytic activity of streptococci and other fastidious microorganisms.

Blood Agar Base, Sheep

Composition per liter:

Pancreatic digest of casein

14.0g

Agar

12.5g

NaCl

5.0g

Peptone

4.5g

Yeast extract

4.5g

Sheep blood, defibrinated

70.0mL

ph 7.3 ± 0.2 at 25°C

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

Preparation: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool the basal medium to 45°–50°C. Aseptically add 70.0mL of sterile, defibrinated sheep blood. Pour into sterile Petri dishes.

Use: For giving improved hemolytic reactions with sheep blood.

Blood Agar Base with Low pH, HiVeg with Blood

Composition per liter:

Agar

15.0g

Plant hydrolysate No. 1

10.0g

Plant infusion

10.0g

NaCl

5.0g

Sheep blood, defibrinated

50.0mL

pH 6. 8 ± 0.2 at 25°C

Source: This medium without blood is available as a premixed powder from HiMedia.

Preparation of Medium: Add components, except sheep blood, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Heat with frequent agitation and boil for 1 min to completely dissolve. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°– 50°C. Aseptically add 50.0mL of sterile, defibrinated sheep blood. Mix thoroughly and pour into sterile Petri dishes.

Use: For the isolation and growth of a wide variety of microorganisms. For the detection of the hemolytic reactions of streptococci and other fastidious microorganisms. The slightly acid pH of this medium enhances distinct hemolytic reactions.

Blood Agar Base with Peptone

Composition per liter:

Agar

15.0g

Beef extract

10.0g

Peptone

10.0g

NaCl

5.0g

pH 7.3 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. 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 use as a base to which blood can be added; for the isolation, cultivation, and detection of hemolytic activity of streptococci and other fastidious microorganisms.

Blood Agar Base with 2.5% Sodium Chloride

Composition per liter:

Beef heart, infusion from

500.0g

NaCl

30.0g

Agar

15.0g

Tryptose

10.0g

pH 6.8 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Heat with frequent agitation and boil for 1 min to completely dissolve. Autoclave for 15 min at 15 psi pressure–121°C. Cool the basal medium to 45°–50°C. Aseptically add sterile, defibrinated blood to a final concentration of 5%. Mix thoroughly and pour into sterile Petri dishes.

Use: For the cultivation of Paracoccus halodenitrificans.

Blood Agar Base with 3.5% Sodium Chloride

Composition per liter:

Beef heart, infusion from

500.0g

NaCl

40.0g

Agar

15.0g

Tryptose

10.0g

pH 6.8 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Heat with frequent agitation and boil for 1 min to completely dissolve. Autoclave for 15 min at 15 psi pressure–121°C. Cool the basal medium to 45°–50°C. Aseptically add sterile, defibrinated blood to a final concentration of 5%. Mix thoroughly and pour into sterile Petri dishes.

Use: For the cultivation of Vibrio costicola.

Blood Agar Base with Special Peptone

Composition per liter:

Agar

15.0g

Beef extract

10.0g

Special peptone

10.0g

NaCl

5.0g

Sheep blood, defibrinated

50.0mL

pH 7.3 ± 0.2 at 25°C

Source: Special peptone (L72) is available from Oxoid Unipath.

Preparation of Medium: Add components, except sheep blood, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Heat with frequent agitation and boil for 1 min to completely dissolve. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°– 50°C. Aseptically add 50.0mL of sterile, defibrinated sheep blood. Mix thoroughly and pour into sterile Petri dishes.

Use: For the isolation, cultivation, and detection of hemolytic activity of streptococci and other fastidious microorganisms.

Blood Agar Base No. 2

(BAM M22)

Composition per 1004.0mL:

Agar

12.0g

Proteose peptone

15.0g

NaCl

5.0g

Yeast extract

5.0g

Liver digest

2.5g

Horse blood, defibrinated

50.0mL

FBP solution

4.0mL

pH 7.4 ± 0.2 at 25°C

FBP Solution:

Composition per 30.0mL:

FeSO4

0.25g

NaHSO3

0.25g

Sodium pyruvate

0.25g

Preparation of FBP Solution: Add components to distilled/deionized water and bring volume to 30.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except horse blood and FBP solution, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 48°C. Aseptically add 50.0mL of sterile horse blood. Mix thoroughly. Aseptically add 4.0mL sterile FBP solution. Mix thoroughly. Pour into sterile Petri dishes in 20.0mL volumes.

Use: For the cultivation of Brucella spp. and other fastidious bacteria.

Blood Agar Base No. 2, HiVeg with Blood

Composition per liter:

Agar

15.0g

Plant peptone No. 3

15.0g

NaCl

5.0g

Yeast extract

5.0g

Plant extract No. 2

2.5g

Horse blood, defibrinated

70.0mL

Selective supplement

2 vials

pH 7.4 ± 0.2 at 25°C

Source: This medium without blood or supplement is available as a premixed powder from HiMedia.

Preparation of Medium: Add components, except horse blood and selective supplement, to distilled/deionized water and bring volume to 930.0mL. 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 70.0mL of sterile horse blood. Mix thoroughly. Aseptically add 2 vials of rehydrated selective supplement. For Brucella spp. use Brucella selective supplement. For Campylobacter spp. use Campylobacter supplement-I (Blaser-Wang), or Campylobacter Supplement II (Butzler), or Campylobacter Supplement III (Skirrow), or Campylobacter Growth Supplement. For streptococci use Strepto supplement. Mix thoroughly. Pour into sterile Petri dishes in 20.0mL volumes.

Use: For the cultivation of Brucella spp., Campylobacter spp., Streptococcus spp., and other fastidious bacteria.

Blood Agar Base No. 2 with 1.2% Agar, HiVeg™

Composition per liter:

Plant peptone No. 3

15.0g

Agar

12.0g

NaCl

5.0g

Yeast extract

5.0g

Plant extract No. 2

2.5g

Horse blood, defibrinated

70.0mL

pH 7.4 ± 0.2 at 25°C

Source: This medium without blood is available as a premixed powder from HiMedia.

Preparation of Medium: Add components, except horse blood and selective supplement, to distilled/deionized water and bring volume to 930.0mL. 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 70.0mL of sterile horse blood. Mix thoroughly. Aseptically add 2 vials of rehydrated selective supplement. For Brucella spp. use Brucella selective supplement. For Campylobacter spp. use Campylobacter supplement-I (Blaser-Wang), or Campylobacter Supplement II (Butzler), or Campylobacter Supplement III (Skirrow), or Campylobacter Growth Supplement. For streptococci use Strepto supplement. Mix thoroughly. Pour into sterile Petri dishes in 20.0mL volumes.

Use: For the cultivation of Brucella spp., Campylobacter spp., Streptococcus spp., and other fastidious bacteria.

Blood Agar, Diphasic

Composition per 800.0mL:

Lean beef, desiccated

25.0g

Agar

10.0g

Neopeptone

10.0g

NaCl

2.5g

Locke solution

200.0mL

Rabbit blood, defibrinated

100.0mL

pH 7.2–7.4 at 25°C

Locke Solution:

Composition per liter:

NaCl

8.0g

Glucose

2.5g

KH2PO4

0.3g

KCl

0.2g

CaCl2·2H2O

0.2g

Preparation of Locke Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add beef to 500.0mL of distilled/deionized water. Let stand for 60 min. Gently heat and bring to 80°C for 5 min. Filter through Whatman #1 filter paper. To filtrate, add remaining components, except Locke solution and rabbit blood. Mix thoroughly. Adjust pH to 7.2–7.4 with NaOH. Autoclave for 20 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add sterile rabbit blood. Mix thoroughly. Aseptically distribute into sterile tubes in 5.0mL volumes. Allow tubes to cool in a slanted position. Immediately prior to inoculation, overlay agar in each tube with 2.0mL of sterile Locke solution.

Use: For the cultivation of Trypanosoma species and Leishmania species.

Blood Agar, Diphasic Base Medium

Composition per 750.0mL:

Beef

25.0g

Agar

10.0g

Neopeptone

10.0g

NaCl

2.5g

pH 7.2–7.4 at 25°C

Preparation of Medium: Trim beef to remove fat. Add 25.0g of lean beef to 250.0mL of distilled/deionized water. Gently heat and bring to boiling. Boil for 2–3 min. Filter through Whatman #2 filter paper. Add agar, neopeptone, and NaCl to filtrate. Bring volume to 750.0mL with distilled/deionized water. Mix thoroughly. Adjust pH to 7.2–7.4. Gently heat a