Christensenella sp

Grow in recommended media (preparation information listed below), at 35°C, in oxygen free nitrogen, and incubate for 5 to 7 days.

The basal medium used consisted of (1 Litter of distilled water): 0.5 g K₂HPO₄, 0.5 g KH₂PO₄, 0.5 g NH₄Cl, 0.4 g KCl, 0.05 g CaCl₂ . 2H₂O, 0.3 g MgCl₂ . 6H₂O, 0.4 g NaCl, 0.25 g HCl-Cysteine, 1 ml trace element mineral solution (composition below) and 1 ml resazurin 0.1% (w/v). 30g of Fastidious Anaerobic Broth and 2g of Tryptone were added to the medium and the pH is adjusted to 6.5.
The medium then was heated to boiling and until the medium turn colorless. The medium is t then cooled at room temperature under a stream of O₂-free nitrogen. The medium is finally sterilized by autoclaving for 20 min at 121 ℃.

After autoclaving, solutions of NaHCO₃ (10%) and Na₂S.9H₂O (2%; pH adjusted) were autoclaved separately and added to the medium (20 mL each) at room temperature. The medium was also supplemented with 1 ml of a filter sterilized (0.22 μm) vitamin solution (composition below). The final pH after adding solutions should be to pH 7.0±0.1.

Element mineral solution composition (Quantity for 1000 ml)
HCl (37%): 6.7 ml
FeCl₂. 4H₂O*: 1500 mg
H₃BO₃ 3: 6 mg
MnCl₂. 4H2O: 100 mg
CoCl₂. 6H2O: 190 mg
ZnCl₂: 70 mg
NiCl₂. 6H2O: 24 mg
CuCl₂. 2H2O: 2 mg
Na₂MoO₄: 36 mg

Vitamin solution composition (Quantity for 100 ml)
Para-amino-benzoic acid: 25 mg
D-biotin: 10 mg
Cyanocobalamin: 0.5 mg
Thiamine-HCl: 25 mg
Riboflavin: 25 mg
Pyridoxal-HCl: 50 mg
DL-pantothenate de calcium: 25 mg
Nicotinic acid (niacine): 25 mg
Folic acid: 10 mg
Lipoïc acid: 25 mg

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