1845 . Nitrososphaera gargensis medium

Equipment needed: Autoclave Filter

Final pH: 8.5

Description: Medium originally described for the cultivation of Nitrososphaera gargensis (Krümmel and Harms, 1982; Hatzenpichler et al., 2008) and modified for the cultivation of Nitrosotenuis aquarius and Nitrosocosmicus hydrocola.

Select a strain (optional):

	Compound	Amount	Unit
	Basal salt stock solution (20x)	50.0	ml
	Trace element solution (Nitrososphaera)	1.0	ml
	FeNaEDTA solution (7.5 mM)	1.0	ml
	Selenite-tungstate solution	1.0	ml
	NH4Cl (1 M)	0.5	ml
1 Add 50 mL of 20x Basal salt stock solution and 4 g CaCO₃ to 950 mL sterile double distilled water and mix all just before use.
2 Autoclave at 121 °C for 20 min.
3 Cool down to room temperature.
4 Add aseptic per liter of 1x basalt salt solution: 1 mL FeNaEDTA (7.5 mM), 1 mL trace elements solution, 1 mL Selenite-tungstate solution, 0.5 mL NH₄Cl solution.
5 Adjust pH to 8.5 with NaOH.

Basal salt stock solution (20x) #

	Compound	Amount	Unit
	KH₂PO₄	1.00	g
	KCl	1.50	g
	Mg(SO₄) x 7 H₂O	7.00	g
	NaCl	11.68	g
1 Prepare with double distilled water.
2 Autoclave at 121 °C for 20 min.
3 Cool down to room temperature.
4 Store in the dark at 4 °C.

Trace element solution (Nitrososphaera) #

1000

	Compound	Amount	Unit
	HCl (12.5M)	8	mL
	H₃BO₃	30	mg
	MnCl₂ x 4 H₂O	100	mg
	CoCl₂ x 6 H₂O	190	mg
	NiCl₂ x 6 H₂O	24	mg
	CuCl₂ x 2 H₂O	2	mg
	ZnSO₄ x 7 H₂O	144	mg
	Na₂MoO₄ x 2 H₂O	36	mg
	Double distilled water	1000	ml
1 This solution contains no iron! Sterilise by autoclaving or filtration (0.2 µm) and store in the dark at 4 °C.

FeNaEDTA solution (7.5 mM) #

1000

	Compound	Amount	Unit
	FeNa-EDTA	2753	mg
	Double distilled water	1000	ml
1 Sterilise by filtration (0.2 µm) and store in the dark at 4 °C.

Selenite-tungstate solution #

1000

Compound	Amount	Unit
NaOH	0.5	g
Na₂SeO₃ x 5 H₂O	3.0	mg
Na₂WO₄ x 2 H₂O	4.0	mg
Distilled water	1000.0	ml

NH4Cl (1 M) #

	Compound	Amount	Unit
	NH₄Cl	2.67	g
	Double distilled water	50.00	ml
1 Dissolve 2.67 g ammonium chloride in 50 mL Double distilled water.
2 Filtrate (0.2 µm).

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1845 . Nitrososphaera gargensis medium

Basal salt stock solution (20x) #

Trace element solution (Nitrososphaera) #

FeNaEDTA solution (7.5 mM) #

Selenite-tungstate solution #

NH4Cl (1 M) #

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