Table 3. Preparation of optimized culture medium for C. bombi.

FP Stock solution(1)
NaCl	2.8 g
KCL	0.4 g
NaH2PO4×H2O	10.0 g
Tryptose Broth (Difco™)	10.0 g
Liver Infusion Broth (Difco™)	2.0 g
H2O bidest: fill to total volume of 1000 ml; set to pH of 5.8
10× FB stock solution(2)
D(-)Fructose	1.8 g
L-Proline	0.289 g
Vitamine B1 hydrochloride	0.0067 g
Folic acid	0.0044 g
FP stock solution to fill to final volume of 100 ml
Heat inactivated fetal bovine serum (hiFBS)(3)
Haemin stock (2 mg/ml)(4)
Haemin-chloride	100.0 mg
NaOH (1 M)	1.25 ml
H2O bidest	23.75 ml
Dissolve by heating to boiling point, cool down and add H2O to final volume of 50 ml
Full medium
FP Stock solution	36.0 ml
10× FB stock solution,	4.0 ml
hiFBS	4.5 ml
Haemin	44.5 µl

⁽¹⁾autoclave and store at 4°C.

⁽²⁾aliquot to 5.0 ml and store at −20°C.

⁽³⁾aliquot to 5.0 ml and store at −20°C.

⁽⁴⁾aliquot to 50 µl.