Chopped Meat Medium with Formate and Fumarate

[所属分类:培养基配方] [发布时间:2020-4-24] [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

Chopped Meat Medium with Formate and Fumarate

培养基配方
Composition per 1230.0mL:
Peptone .................................................................30.0g
K2HPO4 ...................................................................5.0g
Yeast extract.............................................................5.0g
L- Cysteine·HCl·H2O ....................................................0.5g
Chopped meat extract filtrate.............................................1.0L
Chopped meat extract solids............................................200.0mL
Resazurin (0.025% solution) .............................................4.0mL
Formate-fumarate solution...............................................0.05mL
pH 7.0 ± 0.2 at 25°C
Chopped Meat Extract:
Composition per liter:
Beef or horse meat.......................................................500.0g
NaOH (1N solution).......................................................25.0mL
Preparation of Chopped Meat Extract: Use lean beef or horse
meat. Remove fat and connective tissue. Grind. Add meat and NaOH
to distilled/deionized water and bring volume to 1.0L. Gently heat and
bring to boiling while stirring. Cool to 25°C. Remove fat from surface.
Filter. Reserve ground meat particles and filtrate. Add distilled/deion-
ized water to filtrate and bring volume to 1.0L.
Formate-Fumarate Solution:
Composition per 100.0mL:
Sodium formate.............................................................6.0g
Fumaric acid ..............................................................6.0g
Preparation of Formate-Fumarate Solution: Add components
to distilled/deionized water and bring volume to 100.0mL. Adjust pH
to 7.0. Filter sterilize.
Preparation of Medium: To 1.0L of chopped meat extract filtrate,
add the remaining components, except the  L- cysteine·HCl·H2O, formate-
fumarate solution, and chopped meat solids. Mix thoroughly. Gently heat
to boiling. Cool to room temperature. Add the  L- cysteine·HCl·H2O. Ad-
just pH to 7.0. Distribute 1 part chopped meat solids (by volume) and 5
parts of liquid (by volume) into tubes under O2 -free 97% N2 + 3% H2 .
Cap with rubber stoppers and place tubes in a press. Autoclave for 15
min at 15 psi pressure–121°C with fast exhaust. Prior to inoculation, add
0.05mL of formate-fumarate solution to each tube containing approxi-
mately 6.5mL of chopped meat medium.
Use:  For the cultivation and maintenance of Bacteroides ureolyticus
and Wolinella species.