Clostridium botulinum Isolation HiVeg 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 HiVeg Agar

培养基配方
Composition per liter:
Plant hydrolysate............................................................40.0g
Agar ........................................................................20.0g
Na2HPO4 ......................................................................5.0g
Yeast extract.................................................................5.0g
Glucose ......................................................................2.0g
NaCl..........................................................................2.0g
MgSO4 .......................................................................0.01g
Egg yolk emulsion, 50%......................................................100.0mL
Cycloserine solution ........................................................25.0mL
Sulfamethoxazole solution.....................................................4.0mL
Trimethoprim solution ........................................................4.0mL
pH 7.4 ± 0.2 at 25°C
Source: This medium, without egg yolk emulsion, cycloserine solu-
tion, sulfmethoxazole solution, and trimethoprim solution, is available
as a premixed powder (C. botulinum Isolation HiVeg Agar Base) from
HiMedia.
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:  Add components, except egg yolk emul-
sion, cycloserine solution, sulfmethoxazole solution, and trimethoprim
solution, to distilled/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. Aseptically add warmed, sterile
egg yolk emulsion, 50%, and sterile cycloserine solution, sterile sul-
famethoxazole solution, and sterile trimethoprim solution. Mix thor-
oughly. 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.