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Main sol. 78

Internal Solution-ID: S123

This solution is used in 5 mediaand was first defined in medium 78.

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Ground beef
(fat free)
500.0 g 500 -
Distilled water 1000.0 ml - -
NaOH
(1 N)
25.0 ml - -
1 Use lean beef or horse meat. Remove fat and connective tissue before grinding. Mix meat, water and NaOH, then boil for 15 min with stirring. Cool to room temperature, skim fat off surface, and filter, retaining both meat particles and filtrate. To the filtrate add water to a final volume of 1000 ml, and then add:
Casitone 30.0 g 30 -
Yeast extract 5.0 g 5 -
K2HPO4 5.0 g 5 28.707
Resazurin 1.0 mg 1.0e-3 0.004
2 To make medium anoxic bring it to a boil, cool under 100% N2 gas atmosphere, add 0.5 g/l L-cysteine hydrochloride and adjust pH to 7.0. Dispense under 100% N2 gas atmosphere by filling 7 ml medium into anoxic Hungate-type tubes (for strains demanding meat particles put those first into the tube (use 1 part meat particles to 4 or 5 parts fluid)). Autoclave at 121°C for 30 min. For agar slants use 15.0 g agar per 1000.0 ml medium.
L-Cysteine HCl 0.5 g 0.5 3.172
Agar , for solid medium 15.0 g 15 44.599
3 In some cases (as indicated in the catalogue) the addition of Haemin and Vitamin K1 or Vitamin K3 is necessary. Add to 1000 ml of medium after autoclaving:
Haemin solution
optional
10.0 ml - -
Vitamin K1 solution , alternative
optional
10.0 ml - -
Vitamin K3 solution , alternative
optional
10.0 ml - -
78a CHOPPED MEAT MEDIUM FOR TREPONEMA SP.
78 CHOPPED MEAT MEDIUM
78d MEDIUM FOR TREPONEMA PARVUM
209 EUBACTERIUM LENTUM MEDIUM
413 BTU MEDIUM