Clostridium botulinum Isolation Agar (CBI Agar)

[所属分类:培养基配方] [发布时间:2020-5-8] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Clostridium botulinum Isolation Agar
(CBI Agar)

培养基配方
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 components to dis-
tilled/deionized water and bring volume 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 thorough-
ly.
Cycloserine Solution:
Composition per 100.0mL:
Cycloserine.................................................................1.0g
Preparation of Cycloserine Solution: Add cycloserine to dis-
tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.
Filter sterilize.
Sulfamethoxazole Solution:
Composition per 100.0mL:
Sulfamethoxazole............................................................1.9g
Preparation of Sulfamethoxazole Solution: Add sulfamethox-
azole to distilled/deionized water and bring volume to 50.0mL. Add suffi-
cient 10% NaOH to dissolve. Bring volume to 100.0mL with distilled/
deionized water. Mix thoroughly. Filter sterilize.
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 sufficient 0.05N HCl to dissolve. Bring volume to 100.0mL
with distilled/deionized water. Mix thoroughly. 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.
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.
Preparation of Medium:  Aseptically add warmed, sterile egg yolk
emulsion, 50%, and sterile cycloserine solution, sterile sulfamethox-
azole solution, and sterile trimethoprim solution to cooled, sterile 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 from fecal speci-
mens associated with foodborne and infant botulism. Clostridium bot-
ulinum 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 colonies with no opaque zone.