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Clostridium botulinum Isolation Agar (CBI Agar)

[所属分类:培养基配方] [发布时间:2021-11-12] [发布人:网站管理员2] [阅读次数:] [返回]

Clostridium botulinum Isolation Agar

(CBI Agar)

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

Composition per 1033.0mL:

Egg yolk agar base...................................... 900.0mL

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

Cycloserine solution ..................................... 25.0mL

Sulfamethoxazole solution.............................. 4.0mL

Trimethoprim solution .................................... 4.0mL

pH7.4 ± 0.2 at 25°C

Egg Yolk Agar Base:

Composition per 900.0mL:

Pancreatic digest of casein................................40.0g

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

Na2HPO4.............................................................5.0g

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

Glucose ...............................................................2.0g

NaCl....................................................................2.0g

MgSO4·7H2O solution .................................... 0.2mL

Preparation of Egg Yolk Agar Base: Add com

ponents to distilled/deionized water and bring vol

ume to 900.0mL. Mix thoroughly. Gently heat to

boiling. Autoclave for 15 min at 15 psi pressure–

121°C. Cool to 45°–50°C.

MgSO4·7H2O Solution:

Composition per 100.0mL:

MgSO4·7H2O......................................................5.0g

Preparation of MgSO4·7H2O Solution: Add

MgSO4·7H2O to distilled/deionized water and bring

volume to 100.0mL. Mix thoroughly.

Cycloserine Solution:

Composition per 100.0mL:

Cycloserine .........................................................1.0g

Preparation of Cycloserine Solution: Add cy

closerine to distilled/deionized water and bring vol

ume to 100.0mL. Mix thoroughly. Filter sterilize.

Sulfamethoxazole Solution:

Composition per 100.0mL:

Sulfamethoxazole ...............................................1.9g

Preparation of Sulfamethoxazole Solution:

Add sulfamethoxazole to distilled/deionized water

and bring volume to 50.0mL. Add sufficient 10%

NaOH to dissolve. Bring volume to 100.0mL with

distilled/deionized water. Mix thoroughly. Filter ster

ilize.

Trimethoprim Solution:

Composition per 100.0mL:

Trimethoprim......................................................0.1g

Preparation of Trimethoprim Solution: Add

trimethoprim to distilled/deionized water and bring

volume to 50.0mL. Gently heat to 55°C. Add suffi

cient 0.05N HCl to dissolve. Bring volume to

100.0mL with distilled/deionized water. Mix thor

oughly. Filter sterilize.

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.

Preparation of Medium: Aseptically add

warmed, sterile egg yolk emulsion, 50%, and sterile

cycloserine solution, sterile sulfamethoxazole solu

tion, and sterile trimethoprim solution to cooled, ster

ile egg yolk agar base. Mix thoroughly. Pour into

sterile Petri dishes.

Use: For isolation, cultivation, and differentiation

based on lipase activity of Clostridium botulinum

types A, B, and F. Clostridium botulinum types A, B,

and F appear as raised colonies surrounded by an

opaque zone. Other Clostridium species and

Clostridium botulinum type G appear as pinpoint col

onies with no opaque zone.

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Clostridium botulinum Isolation Agar (CBI Agar)