Enrichment Broth, pH 7.3 with Pyruvate

[所属分类:培养基配方] [发布时间:2020-7-7] [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

Enrichment Broth, pH 7.3 with Pyruvate

培养基配方
Composition per liter:
Pancreatic digest of casein..................................................17.0g
NaCl..........................................................................5.0g
Papaic digest of soybean meal.................................................3.0g
Glucose ......................................................................2.5g
K2HPO4 .......................................................................2.5g
Yeast extract.................................................................6.0g
Pyruvate solution...........................................................11.1mL
Nalidixic acid solution......................................................8.0mL
Cycloheximide solution.......................................................5.1mL
Acriflavin·HCl solution ....................................................2.0mL
pH 7.3 ± 0.2 at 25°C
Pyruvate Solution:
Composition per 20.0mL:
Sodium pyruvate...............................................................2.0g
Preparation of Pyruvate Solution: Add sodium pyruvate to dis-
tilled/deionized water and bring volume to 20.0mL. Mix thoroughly.
Filter sterilize.
Cycloheximide Solution:
Composition per 10.0mL:
Cycloheximide.................................................................0.1g
Ethanol, absolute............................................................4.0mL
Preparation of Cycloheximide Solution: Add components to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Filter sterilize.
Caution:  Cycloheximide is toxic. Avoid skin contact or aerosol for-
mation and inhalation.
Nalidixic Acid Solution:
Composition per 10.0mL:
Nalidixic acid...............................................................0.05g
Preparation of Nalidixic Acid Solution: Add nalidixic acid to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Filter sterilize.
Acriflavin·HCl Solution:
Composition per 10.0mL:
Acriflavin·HCl..............................................................0.05g
Preparation of Acriflavin·HCl Solution: Add acriflavin·HCl to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Filter sterilize.
Preparation of Medium: Add components—except pyruvate solu-
tion, cycloheximide solution, nalidixic acid solution, and acriflavin·HCl
solution—to distilled/deionized water and bring volume to 973.8mL.
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 11.1mL
of sterile pyruvate solution. Mix thoroughly. Inoculate medium and in-
cubate at 30°C for 6 hr. Aseptically add 5.1mL of sterile cycloheximide
solution, 8.0mL of sterile nalidixic acid solution, and 2.0mL of sterile
acriflavin·HCl solution. Mix thoroughly.
Use: For the isolation, cultivation, and enrichment of Listeria species
from milk and dairy products.