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Table T13. Compositions of culture media and incubation temperatures for shipboard microbiological studies, Expedition 301.

Culture medium/Components Concentration/
Amount		 Basement
saline		 Gas phase pH Temperature (°C)
Gas Mix Pressure
Methanogen medium 1 (Me1):
MJ H2/CO2 80/20 350 kPa 6.5 20, 37, 55, 70
Acetate 0.1%
NaHCO3 0.1%
Na2S·9H2O 0.05%
Cysteine-HCl 0.05%
Vitamin solution
Resazurin 0.0001%
 
Thermococcales (Tc):
MJ N2
200 kPa 7.0 37, 55, 70, 85
Yeast extract 0.2%
Tryptone 0.2%
Glucose 0.02%
S° 3%
Resazurin 0.0001%
Na2S·9H2O 0.05%
 
Archaeoglobales (Ag):
MJ H2/CO2 80/20 350 kPa 6.5 20, 37, 55, 85
Yeast extract 0.1%
Acetate 0.1%
NaHCO3 0.1%
Na2SO4 0.2%
Na2S·9H2O 0.05%
Vitamin solution
Resazurin 0.0001%
 
Epsilon proteobacteria (Ep):
MJ H2/CO2 80/20 350 kPa 6.5 20, 37, 55, 70
NaNO3 0.1%
Na2S2O3·5H2O 0.1%
S° 3%
NaHCO3 0.1%
FeOOH 100 mM
 
Oligotrophs (Og):
MJ-NP Atmospheric air
100 kPa 7.0 20
Acetate 0.5 mM
Casamino acid 0.005%
Ascorbic acid 0.5 mM
Glucose 2 mM
(NH4)3PO4 0.4 mM
Agar 3%
 
Fermenters (Fm):
MJ N2/CO2/H2 90/5/5 100 kPa 7.0 20
Yeast extract 0.2%
Tryptone 0.2%
Glucose 0.02%
S° 3%
Resazurin 0.0001%
Agar 3%
Monomers (mono):
MM N2/CO2/H2 90/5/5 100 kPa 7.2–7.4 Near in situ temperature
Amino acids mixture 0.01 M
Short chain fatty acids 0.01 M
Organic acid mixture 0.01 M
n-alcohols 0.01 M
Glycerol 0.01 M
Glucose 0.01 M
 
Ammonifex (Amfex):
S-1 H2/CO2 80/20 350 kPa 7.3 Near in situ temperature
NO3– 10 mM
 
Methanogen medium 2 (Me2):






1) Acetate or methanol 10 mM/0.4% S-1 N2/CO2/H2 90/5/5 100 kPa 7.3 Near in situ temperature
2) No additional substrates
H2/CO2 80/20 350 kPa
 
Sulfur (S°) reducers (S-red):






S° 0.5%–1% S-1 N2/CO2 80/20 100 kPa 7.3 Near in situ temperature
Substrates: acetate, PYG, or pectin
 
Denitrifiers (Denit):






CH3COONa·3H2O and Na2SO4 + 0.14 g/0.02 g S-2


7.0 Near in situ temperature
1) NO3– + FeSO4 or FeS 5 mM/10 mM N2/CO2/H2 90/5/5 100 kPa
2) NO3– + 10 mM N2/CO2/H2 90/5/5 100 kPa
Lactate (10 mM) 10 mM
Fumarate (10 mM) 10 mM
Formate (10 mM) 10 mM
 
Iron reducers (IR):






CH3COONa·3H2O, C6H5O7Fe, and Na2SO4 + 0.14 g/30mM/0.02 g S-2


7.0 Near in situ temperature
1) Acetate or lactate 10 mM/10 mM N2/CO2/H2 90/5/5 100 kPa
2) No additional substrates
N2/CO2 80/20 100 kPa
 
Sulfate reducers (SR):






4 mL/L Na2S·9H2O and Na2SO4 + 0.5 M/3.0 g S-2


7.0 Near in situ temperature
1) Lactate and/or acetate 10 mM/10 mM N2/CO2/H2 90/5/5 100 kPa
2) No additional substrates
N2/CO2 80/20 100 kPa

Notes: Compositions of solutions are listed in the following tables: vitamin solution (see Table T14), MJ synthetic seawater (see Table T15); MJ-NP = MJ without nitrogen and phosphate sources; marine medium salts (MM) (see Table T16); S-1 (see Table T17); S-2 (see Table T18). Amino acids mixture = alanine, arginine, asparagine, asparagic acid, cystine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophane, tyrosine, and valine. Short-chain fatty acids (as sodium salts) = formate, acetate, propionate, butyrate, valerate, and capronate. Organic acid mixture = malate, fumarate, succinate, and lactate. n-alcohols = methanol, ethanol, propanol, and butanol. PYG = peptone (0.2%) + yeast extract (0.2%) + glucose (0.05%). See Table T19 for a description of the trace mineral solution added to culture media, Table T20 for a description of the trace element solution used in culture media, Table T21 for a description of the vitamin solution used in S-1 saline solutions and Table T22 for a description of the vitamin solution used in basement saline S-2 solutions.

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