Table T7. Media used for cultivations at 60°C.

Table T7. Media used for cultivations at 60°C.


Medium	Components	Concentration	Comments
Ferm: heterotrophic fermenter	Na₂S	2 mM	Prepare in MSSM under N₂ headspace
	Cystein HCl	2 mM	MPNs pH^60°C = 7.95
	A) glucose, fructose, and mannose	0.2%, wt/vol each	Enrichments pH^60°C = 8.8
	B) xylose and ribose	0.2%, wt/vol each
	C) xylose and ribose + 25 mM NaS₂O₃^2-	0.2%, wt/vol each
H₂/HCO₃^-: acetogenic/methanogenic	Na₂CO₃	15 mM (total 25-30 mM with near-equal amount from sea salt)	Prepare in MSSM under 100% H₂headspace
		15 mM (total 25-30 mM with near-equal amount from sea salt)	MPNs pH^60°C = 7.95 (8.0)
			Enrichments pH^60°C = 8.8
H₂/HCO₃^-/FeIII: iron(III) reducer	Na₂CO₃	15 mM (total 25-30 mM with near-equal amount from sea salt)	Prepare in MSSM under 100% H₂headspace
		15 mM (total 25-30 mM with near-equal amount from sea salt)	MPNs pH^60°C = 7.8
	Fe(III)	90 mM	Enrichments pH^60°C = 8.3

H₂/HCO₃^-/MnIV: manganese(IV) reducer	Na₂CO₃	15 mM (total 25-30 mM with near-equal amount from sea salt)	Prepare in MSSM under 100% H₂headspace
		15 mM (total 25-30 mM with near-equal amount from sea salt)	MPNs pH^60°C = 7.8 (Site 1231 only)
	Mn(IV)(HO,O)_x	15 mM	Enrichments pH^60°C = 7.8 (selected sites)
C-18 lipo: lipolytic microorganism	Cystein HCl	0.15 g	Prepare in MSSM under N₂ headspace
	Yeast extract (total of 0.2%)	0.15%	MPNs pH^60°C = 7.8
	Na₂SO₄	10 mM	Enrichments pH^60°C = 9
	Olive oil	20 ml
SRB: heterotrophic sulfate reducer	Na₂SO₄	20 mM	Prepare in MSSM under N₂ headspace
	Cystein HCl	2 mM	Enumerations pH^60°C = 8.0
	A) Lactate	25 mM	Enrichments pH^60°C = 8.6
	Acetate	25 mM
	Propionate	5 M
	Butyrate	5 mM
	B) Benzoate (only pH = 8)	10 mM	Enumerations pH^60°C = 8.0
Chlor: chlorate reducer	Na₂CO₃ (total bicarbonate/carbonate to be determined later)	15 mM (total 25 mM + equal unknown amount from the salt)	Prepare in MSSM under 100% H₂headspace
	Na chlorate	5 mM
methylamine: methylamine-utilizing methanogens	Na₂S	2 mM	Prepare in MSSM under 100% N₂headspace
methylamine: methylamine-utilizing methanogens	Cystein HCl	2 mM	MPNs pH^25°C = 8.4
	Methylamine	2.5 mM	Enrichments pH^25°C = 8.4
	Trimethylamine	2.5 mM	Site 1229 only; use at 60° and 25°C
CH₄-ace: acetate-utilizing methanogens	Na₂S	2 mM	Prepare in MSSM under 100% N₂headspace
	Cystein HCl	2 mM	MPNs pH^25°C = 8.3
	Acetate	5 mM	Enrichments pH^25°C = 8.3
			Site 1229 only; use at 60° and 25°C

Notes: MSSM = marine sea-salt media (see Table T8). * = Freier et al., 1988. pH^25°C = pH meter was calibrated with the electrode and buffer equilibrated at room temperature. No correction for the sea salt concentration was made. pH^60°C = pH meter, with temperature probe connected, was calibrated with the electrode and buffer equilibrated for 30 min at 60°C. Buffer values were temperature corrected. No correction was made for the sea salt.