Table T4. Media used for cultivation of psychrophiles and mesophiles.
Medium
Components
Concentration
Comments
Mono: monomer Amino acids: alanine, arginine, asparagine, asparagic acid, cystine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophane, tyrosine, and valine 0.01 M Prepare in MM

Short chain fatty acids (sodium salts): formate, acetate, propionate, butyrate, valerate, and capronate 0.01 M

Other organic acids: malate, fumarate, succinate, and lactate 0.01 M

n-alcohols: methanol, ethanol, propanol, and butanol 0.01 M

Glycerol 0.01 M

Glucose 0.01 M




Poly: polymer Chitin, xylane, cellulose, and peptone 0.05% Prepare in MM
Aro: aromatic compounds and long-chain fatty acids Aromatic compounds (sodium salts): salicylate, 3-OH-benzoate,3-OH-benzaldehyde, vanillate, vanillin, p-coumarate, ferulate, sinapinate, syringate, and syringaldehyde 0.05 M Prepare in MM

Fatty acids (sodium salts): C14-C20 0.05 M
Lac: lactate Sodium lactate 5.0 M Prepare in MM




No SO4: sulfate-free Ti(III) citrate 0.5 M Prepare in MM (omit Na2SO4, Na2S, and FeCl2)
Sed: sediment extract N-[2-hydroxyethyl]piperazine-N´-[2-ethane-sulfonic acid] buffer 2.38 g/L Prepare in MM (omit NaHCO3, vitamins, Na2S, and FeCl2) and adjust pH to 7.2-7.4. Use this solution to extract sediment at 80°C (see "Enrichments near In Situ Temperatures" in "Microbiology").

To sediment extract, after heating add the following:


Vitamins, Na2S, and FeCl2


NaHCO3 0.2 g/L
A-bas: autotrophic basalt NaCl 24 g/L Dissolve salts in deionized water, and autoclave 20 min at 121°C

MgCl2·6H2O 10 g/L

KCl 0.5 g/L

Na2SO4 4 g/L

NH4Cl 0.25 g/L

CaCl2 1.5 g/L

KH2PO4 0.2 g/L
AmOx: ammonium oxidizing medium NH4Cl 0.64 g/L Dissolve 10 mg FeCl3·6H2O in 0.3 mL 1-N HCl, add 10 mL deionized water, and then add and dissolve 60 mg EDTA.*
KH2PO4 0.1 g/L

MgCl2·6H2O 0.2 g/L

CaCO3 2.0 g/L

FeCl3·6H2O solution 1 mL/L
NiOx: nitrite-oxidizing medium NaNO2 0.76 g/L NaNO2 is used instead of NH4Cl; the preparation is the same as for AmOx.*
NH4Cl 0.64 g/L

KH2PO4 0.1 g/L

MgCl2·6H2O 0.2 g/L

CaCO3 2.0 g/L

FeCl3·6H2O solution 1 mL/L
Methylo: methylotrophs Agar 15 g/L Dissolve chemicals in 1 L filtered seawater and autoclave for 20 min at 121°C. After cooling to ~60°C, add 5 mL of sterile-filtered methanol. Pour on agar plates; store plate, wrapped in parafilm, at 4°C to minimize evaporation. Plates are inoculated with 0.1-mL aliquots from sample dilution series made using sterile-filtered seawater.

NH4Cl 0.4 g/L

KH2PO4 0.6 g/L

Na2HPO4·7H2O 1.8 g/L

CaCl2 30 mg/L

FeCl3·6H2O 1 mg/L

Notes: MM = marine medium salts solution (see Table T3). EDTA = disodium ethylenediaminetetraacetate. * = for AmOx and NiOx the chemicals were dissolved in 1 L filtered seawater and the pH was adjusted with HCl or NaOH. The solution was sterilized by autoclaving for 20 min at 121°C. After cooling, 1 mL of sterile-filtered iron(III) solution was added. Sterile tubes with 9 mL medium each were inoculated with 1 mL of a nitrite-oxidizing medium (NiOx).