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SFP Agar (Shahidi-Ferguson Perfringens Agar)

[所属分类:培养基配方] [发布时间:2021-8-16] [发布人:网站管理员2] [阅读次数:] [返回]

SFP Agar

(Shahidi-Ferguson Perfringens Agar)

山东拓普生物工程有限公司培养基配方 http://www.topbiol.com

Composition per 2020.0mL:

Basal layer ...........................................................................1010.0mL

Cover layer...........................................................................1010.0mL

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

agnostic Systems and Oxoid Unipath.

Basal Layer:

Composition per 1010.0mL:

Agar ............................................................................................ 20.0g

Tryptose ...................................................................................... 15.0g

Papaic digest of soybean meal...................................................... 5.0g

Yeast extract.................................................................................. 5.0g

Ferric ammonium citrate............................................................... 1.0g

NaHSO3 ........................................................................................ 1.0g

Egg yolk emulsion, 50%........................................................100.0mL

Antibiotic inhibitor ..................................................................10.0mL

pH 7.6 ± 0.2 at 25°C

Egg Yolk Emulsion, 50%:

Composition per 100.0mL:

Chicken egg yolks............................................................................ 11

Whole chicken egg ............................................................................ 1

NaCl (0.9% solution)...............................................................50.0mL

Preparation of Egg Yolk Emulsion, 50%: Soak eggs with 1:100

dilution of saturated mercuric chloride solution for 1 min. Crack eggs

and separate yolks from whites. Mix egg yolks with 1 chicken egg.

Beat to form emulsion. Measure 50.0mL of egg yolk emulsion and add

to 50.0mL of 0.9% NaCl solution. Mix thoroughly. Filter sterilize.

Warm to 45°–50°C.

Antibiotic Inhibitor:

Composition per 10.0mL:

Kanamycin................................................................................ 0.012g

Polymyxin B sulfate .............................................................. 30,000U

Preparation of Antibiotic Inhibitor: Add components to dis

tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.

Filter sterilize.

Preparation of Basal Layer: Add components—except egg yolk

emulsion, 50%, and antibiotic inhibitor—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 egg yolk emulsion, 50%, and antibi

otic inhibitor. Mix thoroughly. Pour into sterile Petri dishes in 10.0mL

volumes.

Cover Layer:

Composition per 1010.0mL:

Agar ............................................................................................ 20.0g

Tryptose ...................................................................................... 15.0g

Papaic digest of soybean meal...................................................... 5.0g

Yeast extract.................................................................................. 5.0g

Ferric ammonium citrate............................................................... 1.0g

NaHSO3 ........................................................................................ 1.0g

Antibiotic inhibitor ..................................................................10.0mL

pH 7.6 ± 0.2 at 25°C

Preparation of Cover Layer: Add components—except antibiotic

inhibitor—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 sterile anti

biotic inhibitor. Mix thoroughly.

Preparation of Medium: Prepare and dispense basal layer into

sterile Petri dishes in 10.0mL volumes. Incubate overnight to dry plates

and test for sterility. Inoculate plates using 0.1mL volume. Spread in

oculum over suface of agar. Aseptically add 10.0mL of cover layer to

each plate. Incubate at 37°C under 90% N2 + 10% CO2.

Use: For the isolation and enumeration of Clostridium perfringens

from foods. Clostridium perfringens appears as black colonies sur

rounded by a precipitate.

S.F.P. HiVeg Agar Base with Egg Yolk and Antibiotics

Composition per liter:

Basal layer..............................................................................500.0mL

Cover layer.............................................................................500.0mL

Basal Layer:

Composition per liter:

Agar ............................................................................................ 20.0g

Plant hydrolysate No. 1............................................................... 15.0g

Yeast extract.................................................................................. 5.0g

Papaic digest of soybean meal ...................................................... 5.0g

NaHSO3 ........................................................................................ 1.0g

Ferric ammonium citrate............................................................... 1.0g

Egg yolk emulsion, 50%........................................................100.0mL

Antibiotic inhibitor ..................................................................10.0mL

pH 7.6 ± 0.2 at 25°C

Source: This medium, without egg yolk emulsion and antibiotic in

hibito, is available as a premixed powder from HiMedia.

Egg Yolk Emulsion, 50%:

Composition per 100.0mL:

Chicken egg yolks............................................................................ 11

Whole chicken egg............................................................................. 1

NaCl (0.9% solution) ...............................................................50.0mL

Preparation of Egg Yolk Emulsion, 50%: Soak eggs with 1:100

dilution of saturated mercuric chloride solution for 1 min. Crack eggs

and separate yolks from whites. Mix egg yolks with 1 chicken egg.

Beat to form emulsion. Measure 50.0mL of egg yolk emulsion and add

to 50.0mL of 0.9% NaCl solution. Mix thoroughly. Filter sterilize.

Warm to 45°–50°C.

Antibiotic Inhibitor:

Composition per 10.0mL:

Kanamycin ................................................................................ 0.012g

Polymyxin B sulfate............................................................... 30,000U

Preparation of Antibiotic Inhibitor: Add components to dis

tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.

Filter sterilize.

Preparation of Basal Layer: Add components—except egg yolk

emulsion, 50%, and antibiotic inhibitor—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 egg yolk emulsion, 50%, and antibi

otic inhibitor. Mix thoroughly. Pour into sterile Petri dishes in 10.0mL

volumes.

Cover Layer:

Composition per 1010.0mL:

Agar ............................................................................................ 20.0g

Tryptose ...................................................................................... 15.0g

Papaic digest of soybean meal ...................................................... 5.0g

Yeast extract.................................................................................. 5.0g

Ferric ammonium citrate............................................................... 1.0g

NaHSO3 ........................................................................................ 1.0g

Antibiotic inhibitor ..................................................................10.0mL

pH 7.6 ± 0.2 at 25°C

Antibiotic Inhibitor:

Composition per 10.0mL:

Kanamycin................................................................................ 0.012g

Polymyxin B sulfate .............................................................. 30,000U

Preparation of Antibiotic Inhibitor: Add components to dis

tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.

Filter sterilize.

Preparation of Cover Layer: Add components—except antibiotic

inhibitor—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 sterile anti

biotic inhibitor. Mix thoroughly.

Preparation of Medium: Prepare and dispense basal layer into

sterile Petri dishes in 10.0mL volumes. Incubate overnight to dry plates

and test for sterility. Inoculate plates using 0.1mL volume. Spread in

oculum over suface of agar. Aseptically add 10.0mL of cover layer to

each plate. Incubate at 37°C under 90% N2 + 10% CO2.

Use: For the isolation and enumeration of Clostridium perfringens

from foods. Clostridium perfringens appears as black colonies sur

rounded by a precipitate.

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SFP Agar (Shahidi-Ferguson Perfringens Agar)