# 909: TREPONEMA DENTICOLA MEDIUM


## Main sol. 909

1000	ml	**Solution A**	
45	ml	**Solution B**	

1. Add 0.45 ml Sol. B to 10 ml Sol. A. Inoculate and pressurize the tubes with H2 to 0.5 bar.


## Solution A

12.5	g	Brain heart infusion broth	
10	g	Trypticase (BBL 11921)	
2.5	g	Yeast extract	
0.5	g	Na-thioglycolate	
1	g	L-Cysteine HCl	
0.25	g	L-Asparagine	
2	g	Glucose	
3	g	Agar	
1000	ml	Distilled water	

1. Boil for 5 min, cool to 50°C while gassing with 80% N2 + 20% CO2. Distribute to anaerobic tubes under the same atmosphere and autoclave.


## Solution B

6	mg	Thiamine pyrophosphate	
2	g	NaHCO3	
25	ml	Distilled water	
20	ml	**Volatile fatty acid solution**	
20	ml	Rabbit serum	

1. Sterilize by filtration, add 20 ml rabbit serum, gas with 80% N2 + 20% CO2.


## Volatile fatty acid solution

100	ml	KOH (0.1 N)	
0.5	ml	Isobutyric acid	
0.5	ml	DL-2-Methylbutyric acid	
0.5	ml	Isovaleric acid	
0.5	ml	Valeric acid