CCVC Medium (Cephalothin Cycloheximide Vancomycin Colistin Medium

[所属分类:培养基配方] [发布时间:2020-4-10] [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

CCVC Medium
(Cephalothin Cycloheximide
Vancomycin Colistin Medium

培养基配方
Composition per liter:
BCYE-alpha base...........................................990.0mL
Antibiotic supplement solution.............................10.0mL
pH 6.9 ± 0.2 at 25°C
Source:  This medium is available as a premixed powder from BD Di-
agnostic Systems.
BCYE-Alpha Base:
Composition per liter:
Agar........................................................15.0g
Yeast extract...............................................10.0g
ACES buffer (2-[(2-amino-2-oxoethyl)-
amino]-ethane sulfonic acid)................................10.0g
Charcoal, activated..........................................2.0g
α-Ketoglutarate.............................................1.0g
Fe4(P2O7)3·9H2O............................................0.25g
L -Cysteine·HCl·H2O solution ............................10.0mL
L -Cysteine·HCl·H2O Solution:
Composition per 10.0mL:
L- Cysteine·HCl·H2O........................................0.4g
Preparation of  L -Cysteine·HCl·H2O Solution: Add  L -cystei-
ne·HCl·H2O to distilled/deionized water and bring volume to 10.0mL.
Mix thoroughly. Filter sterilize.
Preparation of BCYE-Alpha Base: Add components, except  L -
cysteine·HCl·H2O solution, to distilled/deionized water and bring vol-
ume to 990.0mL. Mix thoroughly. Adjust medium to pH 6.9 with 1N
KOH. Heat gently and bring to boiling for 1 min. Autoclave for 15 min
at 15 psi pressure–121°C. Cool to 50°–55°C. Add 4.0mL of  L-
cysteine·HCl·H2O solution. Mix thoroughly.
Antibiotic Supplement Solution:
Composition  per 10.0mL:
Cycloheximide...............................................80.0mg
Colistin....................................................16.0mg
Cephalothin..................................................4.0mg
Vancomycin ..................................................0.5mg
Preparation of Antibiotic Supplement Solution: Add compo-
nents to 10.0mL of distilled/deionized water. Filter sterilize.
Caution:  Cycloheximide is toxic. Avoid skin contact or aerosol for-
mation and inhalation.
Preparation of Medium: To cooled BCYE-alpha base, add
10.0mL sterile antibiotic supplement. Mix thoroughly. Adjust pH to
6.9 with sterile 1N KOH. Pour into sterile Petri dishes with constant
agitation to keep charcoal in suspension.
Use: For the selective isolation and cultivation of Legionella species
from environmental samples.