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SFP Agar (Shahidi-Ferguson Perfringens Agar)

[所属分类:培养基配方] [发布时间:2021-12-16] [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 Diagnostic Systems and Oxoid.

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.

Measure 50.0mL of egg yolk emulsion and add to

50.0mL of 0.9% NaCl solution. Mix thoroughly. Fil

ter 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 com

ponents to distilled/deionized water and bring vol

ume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Basal Layer: Add components—

except egg yolk emulsion, 50%, and antibiotic inhib

itor—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 antibiotic 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 compo

nents—except antibiotic inhibitor—to distilled/

deionized water and bring volume to 1.0L. Mix thor

oughly. Gently heat and bring to boiling. Autoclave

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

50°C. Aseptically add sterile antibiotic inhibitor. Mix

thoroughly.

Preparation of Medium: Prepare and dispense

basal layer into sterile Petri dishes in 10.0mL vol

umes. Incubate overnight to dry plates and test for

sterility. Inoculate plates using 0.1mL volume.

Spread inoculum 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 Clostridi

um perfringens from foods. Clostridium perfringens

appears as black colonies surrounded by a precipi

tate.

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SFP Agar (Shahidi-Ferguson Perfringens Agar)