Chopped Meat Medium for Treponema spp. (DSMZ Medium 78a)

[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 for Treponema spp.
(DSMZ Medium 78a)

培养基配方
Composition per 1055.0mL:
Ground meat, fat free.........................................................500.0g
Pancreatic digest of casein....................................................30.0g
K2HPO4 .........................................................................5.0g
Yeast extract...................................................................5.0g
L -Cysteine·HCl................................................................0.5g
Resazurin .....................................................................1.0mg
Amino acid solution...........................................................50.0mL
NaOH (1N solution)............................................................25.0mL
Vitamin solution...............................................................5.0mL
pH 7.0 ± 0.2 at 25°C
Amino Acid Solution:
Composition per liter:
L -Glutamine....................................................................0.7g
L -Histidine....................................................................0.6g
L -Serine.......................................................................0.5g
Preparation of Amino Acid Solution:  Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Vitamin Solution:
Composition per liter:
Vitamin B 12 .................................................................50.0mg
Pantothenic acid..............................................................50.0mg
Riboflavin....................................................................50.0mg
α-Lipoic acid................................................................50.0mg
p-Aminobenzoic acid...........................................................50.0mg
Thiamine-HCl·2H2O............................................................50.0mg
Nicotinic acid................................................................25.0mg
Nicotine amide................................................................25.0mg
Biotin........................................................................20.0mg
Folic acid ...................................................................20.0mg
Pyridoxamine-HCl..............................................................10.0mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Filter ster-
ilize.
Preparation of Medium: Use lean beef or horse meat. Remove fat
and connective tissue. Grind finely. Add ground meat and 25.0mL of
NaOH solution to distilled/deionized water and bring volume to
1025.0mL. Gently heat and bring to boiling. Continue boiling for 15
min without stirring. Cool to room temperature. Remove fat from sur-
face. Filter and retain both meat particles and filtrate. Adjust volume of
filtrate to 1.0L with distilled/deionized water. Add pancreatic digest of
casein, K2HPO4 , yeast extract, and resazurin. Gently heat and bring to
boiling. Boil for 1–2 min. Add  L -cysteine·HCl. Mix thoroughly. Auto-
clave for 30 min at 15 psi pressure–121°C. Aseptically add amino acid
and vitamin solutions. Mix thoroughly. Aseptically distribute 7.0mL
amounts into tubes that contain meat particles (1 part meat particles to
5 parts fluid).
Use: For the cultivation of Treponema brennaborense.