Overview
- External ontology
- mediadive.medium:322 (source link unavailable) — METHANOSPHAERA MEDIUM
- Applications
-
- Microbial cultivation
- pH range
- {'min': 6.7, 'max': 6.9}
- Notes
- Source: DSMZ | Link: https://www.dsmz.de/microorganisms/medium/pdf/DSMZ_Medium322.pdf
Composition
flowchart LR
CultureMech_001424["methanosphaera_medium"]
ing0_Trypticase_peptone["Trypticase peptone"]
CultureMech_001424 --> ing0_Trypticase_peptone
ing1_Yeast_extract["Yeast extract"]
CultureMech_001424 --> ing1_Yeast_extract
ing2_Na_acetate["Na-acetate"]
CultureMech_001424 --> ing2_Na_acetate
CHEBI_32954["CHEBI:32954"]
ing2_Na_acetate -.-> CHEBI_32954
ing3_Na_formate["Na-formate"]
CultureMech_001424 --> ing3_Na_formate
CHEBI_62965["CHEBI:62965"]
ing3_Na_formate -.-> CHEBI_62965
ing4_Na2SeO4["Na2SeO4"]
CultureMech_001424 --> ing4_Na2SeO4
CHEBI_77775["CHEBI:77775"]
ing4_Na2SeO4 -.-> CHEBI_77775
ing5_NiCl2_x_6_H2O["NiCl2 x 6 H2O"]
CultureMech_001424 --> ing5_NiCl2_x_6_H2O
CHEBI_34887["CHEBI:34887"]
ing5_NiCl2_x_6_H2O -.-> CHEBI_34887
ing6_K2HPO4["K2HPO4"]
CultureMech_001424 --> ing6_K2HPO4
CHEBI_131527["CHEBI:131527"]
ing6_K2HPO4 -.-> CHEBI_131527
ing7_KH2PO4["KH2PO4"]
CultureMech_001424 --> ing7_KH2PO4
CHEBI_63036["CHEBI:63036"]
ing7_KH2PO4 -.-> CHEBI_63036
ing8__NH4_2SO4["(NH4)2SO4"]
CultureMech_001424 --> ing8__NH4_2SO4
CHEBI_62946["CHEBI:62946"]
ing8__NH4_2SO4 -.-> CHEBI_62946
ing9_NH4Cl["NH4Cl"]
CultureMech_001424 --> ing9_NH4Cl
CHEBI_31206["CHEBI:31206"]
ing9_NH4Cl -.-> CHEBI_31206
ing10_NaCl["NaCl"]
CultureMech_001424 --> ing10_NaCl
CHEBI_26710["CHEBI:26710"]
ing10_NaCl -.-> CHEBI_26710
ing11_MgSO4_x_7_H2O["MgSO4 x 7 H2O"]
CultureMech_001424 --> ing11_MgSO4_x_7_H2O
CHEBI_31795["CHEBI:31795"]
ing11_MgSO4_x_7_H2O -.-> CHEBI_31795
ing12_CaCl2_x_2_H2O["CaCl2 x 2 H2O"]
CultureMech_001424 --> ing12_CaCl2_x_2_H2O
CHEBI_86158["CHEBI:86158"]
ing12_CaCl2_x_2_H2O -.-> CHEBI_86158
ing13_Sodium_resazurin["Sodium resazurin"]
CultureMech_001424 --> ing13_Sodium_resazurin
CHEBI_8806["CHEBI:8806"]
ing13_Sodium_resazurin -.-> CHEBI_8806
ing14_NaHCO3["NaHCO3"]
CultureMech_001424 --> ing14_NaHCO3
CHEBI_32139["CHEBI:32139"]
ing14_NaHCO3 -.-> CHEBI_32139
ing15_Methanol["Methanol"]
CultureMech_001424 --> ing15_Methanol
CHEBI_17790["CHEBI:17790"]
ing15_Methanol -.-> CHEBI_17790
ing16_DL_Dithiothreitol["DL-Dithiothreitol"]
CultureMech_001424 --> ing16_DL_Dithiothreitol
CHEBI_18320["CHEBI:18320"]
ing16_DL_Dithiothreitol -.-> CHEBI_18320
ing17_Nitrilotriacetic_acid["Nitrilotriacetic acid"]
CultureMech_001424 --> ing17_Nitrilotriacetic_acid
CHEBI_44557["CHEBI:44557"]
ing17_Nitrilotriacetic_acid -.-> CHEBI_44557
ing18_MnSO4_x_H2O["MnSO4 x H2O"]
CultureMech_001424 --> ing18_MnSO4_x_H2O
CHEBI_86364["CHEBI:86364"]
ing18_MnSO4_x_H2O -.-> CHEBI_86364
ing19_FeSO4_x_7_H2O["FeSO4 x 7 H2O"]
CultureMech_001424 --> ing19_FeSO4_x_7_H2O
CHEBI_75836["CHEBI:75836"]
ing19_FeSO4_x_7_H2O -.-> CHEBI_75836
ing20_CoSO4_x_7_H2O["CoSO4 x 7 H2O"]
CultureMech_001424 --> ing20_CoSO4_x_7_H2O
CHEBI_91244["CHEBI:91244"]
ing20_CoSO4_x_7_H2O -.-> CHEBI_91244
ing21_ZnSO4_x_7_H2O["ZnSO4 x 7 H2O"]
CultureMech_001424 --> ing21_ZnSO4_x_7_H2O
CHEBI_32312["CHEBI:32312"]
ing21_ZnSO4_x_7_H2O -.-> CHEBI_32312
ing22_CuSO4_x_5_H2O["CuSO4 x 5 H2O"]
CultureMech_001424 --> ing22_CuSO4_x_5_H2O
CHEBI_31440["CHEBI:31440"]
ing22_CuSO4_x_5_H2O -.-> CHEBI_31440
ing23_AlK_SO4_2_x_12_H2O["AlK(SO4)2 x 12 H2O"]
CultureMech_001424 --> ing23_AlK_SO4_2_x_12_H2O
CHEBI_86465["CHEBI:86465"]
ing23_AlK_SO4_2_x_12_H2O -.-> CHEBI_86465
ing24_H3BO3["H3BO3"]
CultureMech_001424 --> ing24_H3BO3
CHEBI_33118["CHEBI:33118"]
ing24_H3BO3 -.-> CHEBI_33118
ing25_Na2MoO4_x_2_H2O["Na2MoO4 x 2 H2O"]
CultureMech_001424 --> ing25_Na2MoO4_x_2_H2O
CHEBI_75213["CHEBI:75213"]
ing25_Na2MoO4_x_2_H2O -.-> CHEBI_75213
ing26_Na2SeO3_x_5_H2O["Na2SeO3 x 5 H2O"]
CultureMech_001424 --> ing26_Na2SeO3_x_5_H2O
CHEBI_131361["CHEBI:131361"]
ing26_Na2SeO3_x_5_H2O -.-> CHEBI_131361
ing27_Na2WO4_x_2_H2O["Na2WO4 x 2 H2O"]
CultureMech_001424 --> ing27_Na2WO4_x_2_H2O
CHEBI_63939["CHEBI:63939"]
ing27_Na2WO4_x_2_H2O -.-> CHEBI_63939
ing28_Biotin["Biotin"]
CultureMech_001424 --> ing28_Biotin
CHEBI_15956["CHEBI:15956"]
ing28_Biotin -.-> CHEBI_15956
ing29_Folic_acid["Folic acid"]
CultureMech_001424 --> ing29_Folic_acid
CHEBI_27470["CHEBI:27470"]
ing29_Folic_acid -.-> CHEBI_27470
CultureMech_001424 --> more["...9 more"]
Ingredients (39)
| # | Ingredient | Concentration | Ontology | Notes |
|---|---|---|---|---|
| 1 | Trypticase peptone | 1.94553 G_PER_L | ||
| 2 | Yeast extract | 1.94553 G_PER_L | ||
| 3 |
Na-acetate
C2H3O2.Na 82.034 g/mol
|
0.486381 G_PER_L |
CHEBI:32954
sodium acetate
|
|
| 4 |
Na-formate
CHO2.Na 68.007 g/mol
|
0.486381 G_PER_L |
CHEBI:62965
sodium formate
|
|
| 5 |
Na2SeO4
2Na.O4Se 188.936 g/mol
|
0.00184825 G_PER_L |
CHEBI:77775
sodium selenate
|
|
| 6 |
NiCl2 x 6 H2O
Cl2Ni 129.599 g/mol
|
0.030680934 G_PER_L |
CHEBI:34887
nickel dichloride
|
[Merged 2 duplicates: 0.000680934, 0.03] |
| 7 |
K2HPO4
HO4P.2K 174.174 g/mol
|
0.583658 G_PER_L |
CHEBI:131527
dipotassium hydrogen phosphate
|
|
| 8 |
KH2PO4
H2O4P.K 136.084 g/mol
|
2.72374 G_PER_L |
CHEBI:63036
potassium dihydrogen phosphate
|
|
| 9 |
(NH4)2SO4
2H4N.O4S 132.141 g/mol
|
0.291829 G_PER_L |
CHEBI:62946
ammonium sulfate
|
|
| 10 |
NH4Cl
Cl.H4N 53.492 g/mol
|
0.972763 G_PER_L |
CHEBI:31206
ammonium chloride
|
|
| 11 |
NaCl
Cl.Na 58.443 g/mol
|
1.583658 G_PER_L |
CHEBI:26710
sodium chloride
|
[Merged 2 duplicates: 0.583658, 1.0] |
| 12 |
MgSO4 x 7 H2O
7H2O.Mg.O4S 246.473 g/mol
|
3.145914 G_PER_L |
CHEBI:31795
magnesium sulfate heptahydrate
|
[Merged 2 duplicates: 0.145914, 3.0] |
| 13 |
CaCl2 x 2 H2O
Ca.2Cl.2H2O 147.014 g/mol
|
0.177821 G_PER_L |
CHEBI:86158
calcium chloride dihydrate
|
[Merged 2 duplicates: 0.077821, 0.1] |
| 14 |
Sodium resazurin
C12H7NO4 229.191 g/mol
|
0.000486381 G_PER_L |
CHEBI:8806
Resazurin
|
|
| 15 |
NaHCO3
CHO3.Na 84.006 g/mol
|
3.89105 G_PER_L |
CHEBI:32139
sodium hydrogencarbonate
|
|
| 16 |
Methanol
CH4O 32.042 g/mol
|
7.70428 G_PER_L |
CHEBI:17790
methanol
|
|
| 17 |
DL-Dithiothreitol
C4H10O2S2 154.256 g/mol
|
0.486381 G_PER_L |
CHEBI:18320
1,4-dithiothreitol
|
|
| 18 |
Nitrilotriacetic acid
C6H9NO6 191.139 g/mol
|
1.5 G_PER_L |
CHEBI:44557
nitrilotriacetic acid
|
|
| 19 |
MnSO4 x H2O
H2O.Mn.O4S 169.016 g/mol
|
0.5 G_PER_L |
CHEBI:86364
manganese(II) sulfate monohydrate
|
|
| 20 |
FeSO4 x 7 H2O
Fe.7H2O.O4S 278.013 g/mol
|
1.1 G_PER_L |
CHEBI:75836
iron(2+) sulfate heptahydrate
|
[Merged 2 duplicates: 0.1, 1.0] |
| 21 |
CoSO4 x 7 H2O
Co.7H2O.O4S 281.101 g/mol
|
0.18 G_PER_L |
CHEBI:91244
cobalt(2+) sulfate heptahydrate
|
|
| 22 |
ZnSO4 x 7 H2O
7H2O.O4S.Zn 287.558 g/mol
|
0.18 G_PER_L |
CHEBI:32312
zinc sulfate heptahydrate
|
|
| 23 |
CuSO4 x 5 H2O
Cu.5H2O.O4S 249.684 g/mol
|
0.01 G_PER_L |
CHEBI:31440
copper(II) sulfate pentahydrate
|
|
| 24 |
AlK(SO4)2 x 12 H2O
Al.12H2O.K.2O4S 474.386 g/mol
|
0.02 G_PER_L |
CHEBI:86465
potassium aluminium sulfate dodecahydrate
|
|
| 25 |
H3BO3
H3BO3 61.833 g/mol
|
0.01 G_PER_L |
CHEBI:33118
boric acid
|
|
| 26 |
Na2MoO4 x 2 H2O
2H2O.MoO4.2Na 241.946 g/mol
|
0.01 G_PER_L |
CHEBI:75213
sodium molybdate dihydrate
|
|
| 27 |
Na2SeO3 x 5 H2O
5H2O.2Na.O3Se 263.012 g/mol
|
0.0003 G_PER_L |
CHEBI:131361
disodium selenite pentahydrate
|
|
| 28 |
Na2WO4 x 2 H2O
2H2O.2Na.O4W 329.846 g/mol
|
0.0004 G_PER_L |
CHEBI:63939
sodium tungstate dihydrate
|
|
| 29 |
Biotin
C10H16N2O3S 244.316 g/mol
|
0.02 G_PER_L |
CHEBI:15956
biotin
|
|
| 30 |
Folic acid
C19H19N7O6 441.404 g/mol
|
0.02 G_PER_L |
CHEBI:27470
folic acid
|
|
| 31 |
Pyridoxine hydrochloride
C8H11NO3.HCl 205.641 g/mol
|
0.1 G_PER_L |
CHEBI:30961
pyridoxine hydrochloride
|
|
| 32 |
Thiamine HCl
C12H17N4OS.Cl.HCl 337.276 g/mol
|
0.05 G_PER_L |
CHEBI:49105
thiamine hydrochloride
|
|
| 33 |
Riboflavin
C17H20N4O6 376.369 g/mol
|
0.05 G_PER_L |
CHEBI:17015
riboflavin
|
|
| 34 |
Nicotinic acid
C6H5NO2 123.111 g/mol
|
0.05 G_PER_L |
CHEBI:15940
nicotinic acid
|
|
| 35 |
Calcium D-(+)-pantothenate
2C9H16NO5.Ca 476.536 g/mol
|
0.05 G_PER_L |
CHEBI:31345
Calcium pantothenate
|
|
| 36 | Vitamin B12 | 0.001 G_PER_L |
CHEBI:176843
vitamin B12
|
|
| 37 |
p-Aminobenzoic acid
C7H7NO2 137.138 g/mol
|
0.05 G_PER_L |
CHEBI:30753
4-aminobenzoic acid
|
|
| 38 |
(DL)-alpha-Lipoic acid
C8H14O2S2 206.332 g/mol
|
0.05 G_PER_L |
CHEBI:16494
lipoic acid
|
|
| 39 |
H2SO4
H2O4S 98.079 g/mol
|
1000 G_PER_L |
CHEBI:26836
sulfuric acid
|
Preparation
- Dissolve ingredients except bicarbonate, methanol, and dithiothreitol (DTT), then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve bicarbonate, adjust pH of medium to 6.8 - 7.0, then dispense medium under 80% H2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Add methanol (50% v/v solution), vitamins, and DTT from sterile anoxic stock solutions prepared under 100% N2 gas. Vitamins and DTT should be sterilized by filtration. Prior to use check pH of complete medium and adjust to 6.7 - 6.9, if necessary.
- After inoculation add sterile 80% H2 and 20% CO2 gas mixture to 1 bar overpressure.
- Rumen fluid from cow or sheep (obtained from fistulated animals or abattoir refuse) is filtered through muslin, autoclaved at 121°C for 15 min and then centrifuged at 27,000 g for 20 min. The supernatant is made anoxic by sparging with 100% N2 gas for 15 min, dispensed under same gas atmosphere into anoxic serum vials to 30% of volume and then stored frozen at -20°C.
- First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH.
- The ferrous sulfate solution is not stable and should be freshly prepared.
Target organisms
No target organisms are recorded.
Provenance
| Timestamp | Curator | Action | Notes |
|---|---|---|---|
| 2026-01-26T17:53:10.791668Z | mediadive-import | Imported from MediaDive | Source: DSMZ, ID: 322 |
| 2026-02-06T05:10:54.086127Z | invalid-chebi-removal | Removed 1 invalid CHEBI ID(s) | Removed CHEBI IDs with invalid format (7+ digits) from MicrobeMediaParam source data |
| 2026-03-13T03:09:42.393207Z | data-quality-cleanup-v1.0 | Data quality fixes | Merged 5 duplicate ingredient(s) |
| 2026-03-13T06:19:04.031120Z | culturemech-id-assigner-v1.0 | Assigned CultureMech ID | Assigned stable identifier: CultureMech:001424 |
| 2026-03-17T06:30:54.000000Z | mediaingredientmech-enrichment-v1.0 | Added MediaIngredientMech links | Linked ingredients to MediaIngredientMech identifiers |
| 2026-03-17T19:45:16.013950+00:00 | normalize_media_names_to_snake_case.py | NAME_NORMALIZED | Normalized name from "METHANOSPHAERA MEDIUM" to "methanosphaera_medium" for consistent matching |
| 2026-05-20T06:01:56.454819+00:00 | migrate_ph_range.py | MIGRATED_PH_RANGE | ph_range '6.7-6.9' -> {'min': 6.7, 'max': 6.9} |
| 2026-06-05T07:41:32.618637+00:00 | mim-legacy-to-chebi-migration-v1.0 | Refreshed legacy MediaIngredientMech links to CHEBI keying | Replaced 34 legacy mediaingredientmech_term link(s) with mediaingredientmech_chebi_term (id-safe: ingredient's own CHEBI + label). MIM deprecated the MediaIngredientMech:NNNNNN id scheme. |
| 2026-06-10T07:37:35.487537+00:00 | primary-term-regrounding-v1.0 | Re-grounded wrong primary term id (OAK-verified) | 1 primary term(s); ingredient name resolved by OAK sqlite:obo:chebi. |
| 2026-06-10T07:47:07.374608+00:00 | primary-term-label-canonicalization-v1.0 | Canonicalized primary term label to OBO canonical (OAK) | 8 primary term(s); ingredient name resolved by OAK sqlite:obo:chebi. |