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SFP Agar (Shahidi-Ferguson Perfringens Agar)

[所屬分類:培養基配方] [發布時間:2021-12-16] [發布人:網站管理員2] [閱讀次數:] [返回]
SFP Agar
(Shahidi-Ferguson Perfringens Agar)

山東拓普生物工程有限公司 培養基配方 //ljxinglong.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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