Chopped Meat Medium, Modified with Formate and Fumarate

[所属分类:培养基配方] [发布时间:2020-4-26] [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,
Modified with Formate and Fumarate

培养基配方
Composition per 1230.0mL:
Pancreatic digest of casein......................................................30.0g
Peptone..........................................................................30.0g
Agar ............................................................................20.0g
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
Hemin solution..................................................................10.0mL
Resazurin (0.025% solution)......................................................4.0mL
Formate-fumarate solution.......................................................0.25mL
Vitamin K 1 solution ............................................................0.2mL
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.
Hemin Solution:
Composition per 100.0mL:
Hemin............................................................................0.05g
NaOH (1N solution)...............................................................1.0mL
Preparation of Hemin Solution: Add components to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly.
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.
Vitamin K 1 Solution:
Composition per 30.0mL:
Ethanol (95% solution)............................................................30.0mL
Vitamin K 1 ......................................................................0.15mL
Preparation of Vitamin K 1 Solution: Mix components. Store so-
lution protected from light at 5°C. Discard after one month.
Preparation of Medium: To 1.0L of chopped meat extract filtrate,
add the remaining components, except the  L- cysteine·HCl·H2O, hemin
solution, vitamin K 1 solution, formate-fumarate solution, and chopped
meat solids. Mix thoroughly. Gently heat to boiling. Cool to room tem-
perature. Add the  L- cysteine·HCl·H2O, hemin solution, and vitamin K 1
solution. Adjust 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 0.25mL of formate-fumarate solution to each tube
containing approximately 5.0mL of chopped meat medium, modified.
Use:  For the cultivation and maintenance of Bacteroides gracilis,
Bacteroides ureolyticus, Campylobacter mucosalis, and Wolinella suc-
cinogenes.