rumen_fluid_medium (CultureMech:010214)

Overview

External ontology

TOGO:M802 — Rumen Fluid Medium

Applications

Microbial cultivation

pH

6.5

Notes

TOGO M802 preserves JCM Medium 773 as Rumen Fluid Medium after the JCM 773 page became unavailable. The JCM formula contains 660.0 ml distilled water, 0.13 g MgSO4 x 7H2O, 0.6 g NaCl, 8.0 mg CaCl2 x 2H2O, 0.3 g KH2PO4, 0.3 g K2HPO4, 1.0 mg resazurin, 0.5 g Na2S x 9H2O, 2.0 mg FeSO4 x 7H2O, 2.0 g NaHCO3, 0.3 g (NH4)2SO4, 1.0 g Yeast extract from BD-Difco, 1.0 g Trypticase peptone from BD-BBL, 0.5 g L-Cysteine HCl H2O, 300.0 ml clarified rumen fluid from JCM Medium 266, 10.0 ml Trace minerals from JCM Medium 151, 10.0 ml Trace vitamins from JCM Medium 197, and H2-CO2/N2 anaerobic gas handling. JCM Medium 773 omits the JCM Medium 266 fatty acid mixture and adjusts the medium to pH 6.5.

Composition

flowchart LR
    CultureMech_010214["rumen_fluid_medium"]
    ing0_Distilled_water["Distilled water"]
    CultureMech_010214 --> ing0_Distilled_water
    CHEBI_15377["CHEBI:15377"]
    ing0_Distilled_water -.-> CHEBI_15377
    ing1_MgSO4_x_7H2O["MgSO4 x 7H2O"]
    CultureMech_010214 --> ing1_MgSO4_x_7H2O
    CHEBI_31795["CHEBI:31795"]
    ing1_MgSO4_x_7H2O -.-> CHEBI_31795
    ing2_NaCl["NaCl"]
    CultureMech_010214 --> ing2_NaCl
    CHEBI_26710["CHEBI:26710"]
    ing2_NaCl -.-> CHEBI_26710
    ing3_CaCl2_x_2H2O["CaCl2 x 2H2O"]
    CultureMech_010214 --> ing3_CaCl2_x_2H2O
    CHEBI_86158["CHEBI:86158"]
    ing3_CaCl2_x_2H2O -.-> CHEBI_86158
    ing4_KH2PO4["KH2PO4"]
    CultureMech_010214 --> ing4_KH2PO4
    CHEBI_63036["CHEBI:63036"]
    ing4_KH2PO4 -.-> CHEBI_63036
    ing5_K2HPO4["K2HPO4"]
    CultureMech_010214 --> ing5_K2HPO4
    CHEBI_131527["CHEBI:131527"]
    ing5_K2HPO4 -.-> CHEBI_131527
    ing6_Resazurin["Resazurin"]
    CultureMech_010214 --> ing6_Resazurin
    CHEBI_8806["CHEBI:8806"]
    ing6_Resazurin -.-> CHEBI_8806
    ing7_Na2S_x_9H2O["Na2S x 9H2O"]
    CultureMech_010214 --> ing7_Na2S_x_9H2O
    CHEBI_76209["CHEBI:76209"]
    ing7_Na2S_x_9H2O -.-> CHEBI_76209
    ing8_FeSO4_x_7H2O["FeSO4 x 7H2O"]
    CultureMech_010214 --> ing8_FeSO4_x_7H2O
    CHEBI_75836["CHEBI:75836"]
    ing8_FeSO4_x_7H2O -.-> CHEBI_75836
    ing9_NaHCO3["NaHCO3"]
    CultureMech_010214 --> ing9_NaHCO3
    CHEBI_32139["CHEBI:32139"]
    ing9_NaHCO3 -.-> CHEBI_32139
    ing10__NH4_2SO4["(NH4)2SO4"]
    CultureMech_010214 --> ing10__NH4_2SO4
    CHEBI_62946["CHEBI:62946"]
    ing10__NH4_2SO4 -.-> CHEBI_62946
    ing11_Yeast_extract__BD_Difco_["Yeast extract (BD-Difco)"]
    CultureMech_010214 --> ing11_Yeast_extract__BD_Difco_
    ing12_Trypticase_peptone__BD_BBL_["Trypticase peptone (BD-BBL)"]
    CultureMech_010214 --> ing12_Trypticase_peptone__BD_BBL_
    ing13_L_Cysteine_HCl_H2O["L-Cysteine HCl H2O"]
    CultureMech_010214 --> ing13_L_Cysteine_HCl_H2O
    CHEBI_91248["CHEBI:91248"]
    ing13_L_Cysteine_HCl_H2O -.-> CHEBI_91248
    ing14_Carbon_dioxide_gas["Carbon dioxide gas"]
    CultureMech_010214 --> ing14_Carbon_dioxide_gas
    CHEBI_16526["CHEBI:16526"]
    ing14_Carbon_dioxide_gas -.-> CHEBI_16526
    ing15_N2["N2"]
    CultureMech_010214 --> ing15_N2
    CHEBI_17997["CHEBI:17997"]
    ing15_N2 -.-> CHEBI_17997
    ing16_Hydrogen_gas["Hydrogen gas"]
    CultureMech_010214 --> ing16_Hydrogen_gas
    CHEBI_18276["CHEBI:18276"]
    ing16_Hydrogen_gas -.-> CHEBI_18276

Ingredients (17)

#	Ingredient	Concentration	Ontology	Notes
1	Distilled water H2O 18.015 g/mol	660.0 ML_PER_L	CHEBI:15377 water	JCM Medium 773 / TOGO M802 lists 660.0 ml/L Distilled water.
2	MgSO4 x 7H2O 7H2O.Mg.O4S 246.473 g/mol	0.13 G_PER_L	CHEBI:31795 magnesium sulfate heptahydrate	JCM Medium 773 / TOGO M802 lists 0.13 g/L MgSO4 x 7H2O.
3	NaCl Cl.Na 58.443 g/mol	0.6 G_PER_L	CHEBI:26710 sodium chloride	JCM Medium 773 / TOGO M802 lists 0.6 g/L NaCl.
4	CaCl2 x 2H2O Ca.2Cl.2H2O 147.014 g/mol	8.0 MG_PER_L	CHEBI:86158 calcium chloride dihydrate	JCM Medium 773 / TOGO M802 lists 8.0 mg/L CaCl2 x 2H2O.
5	KH2PO4 H2O4P.K 136.084 g/mol	0.3 G_PER_L	CHEBI:63036 potassium dihydrogen phosphate	JCM Medium 773 / TOGO M802 lists 0.3 g/L KH2PO4.
6	K2HPO4 HO4P.2K 174.174 g/mol	0.3 G_PER_L	CHEBI:131527 dipotassium hydrogen phosphate	JCM Medium 773 / TOGO M802 lists 0.3 g/L K2HPO4.
7	Resazurin C12H7NO4 229.191 g/mol	1.0 MG_PER_L	CHEBI:8806 Resazurin	JCM Medium 773 / TOGO M802 lists 1.0 mg/L Resazurin.
8	Na2S x 9H2O 9H2O.2Na.S 240.182 g/mol	0.5 G_PER_L	CHEBI:76209 sodium sulfide nonahydrate	JCM Medium 773 / TOGO M802 lists 0.5 g/L Na2S x 9H2O.
9	FeSO4 x 7H2O Fe.7H2O.O4S 278.013 g/mol	2.0 MG_PER_L	CHEBI:75836 iron(2+) sulfate heptahydrate	JCM Medium 773 / TOGO M802 lists 2.0 mg/L FeSO4 x 7H2O.
10	NaHCO3 CHO3.Na 84.006 g/mol	2.0 G_PER_L	CHEBI:32139 sodium hydrogencarbonate	JCM Medium 773 / TOGO M802 lists 2.0 g/L NaHCO3.
11	(NH4)2SO4 2H4N.O4S 132.141 g/mol	0.3 G_PER_L	CHEBI:62946 ammonium sulfate	JCM Medium 773 / TOGO M802 lists 0.3 g/L (NH4)2SO4.
12	Yeast extract (BD-Difco)	1.0 G_PER_L	FOODON:03315426 yeast extract	JCM Medium 773 / TOGO M802 lists 1.0 g/L Yeast extract (BD-Difco).
13	Trypticase peptone (BD-BBL)	1.0 G_PER_L		JCM Medium 773 / TOGO M802 lists 1.0 g/L Trypticase peptone (BD-BBL).
14	L-Cysteine HCl H2O C3H7NO2S.H2O.HCl 175.637 g/mol	0.5 G_PER_L	CHEBI:91248 L-cysteine hydrochloride hydrate	JCM Medium 773 / TOGO M802 lists 0.5 g/L L-Cysteine HCl H2O.
15	Carbon dioxide gas CO2 44.009 g/mol	variable VARIABLE	CHEBI:16526 carbon dioxide	JCM Medium 773 / TOGO M802 uses Carbon dioxide gas as a gas-phase component.
16	N2 N2 28.014 g/mol	variable VARIABLE	CHEBI:17997 dinitrogen	JCM Medium 773 / TOGO M802 uses N2 as a gas-phase component.
17	Hydrogen gas H2 2.016 g/mol	variable VARIABLE	CHEBI:18276 dihydrogen	JCM Medium 773 / TOGO M802 uses Hydrogen gas as a gas-phase component.

Preparation

Mix all ingredients except NaHCO3, L-Cysteine HCl H2O, and Na2S x 9H2O, then adjust the pH to 6.5. Bring the medium to a boil for 5-10 seconds, cool under H2-CO2 (80:20, v/v), add NaHCO3, dispense under H2-CO2, seal, and autoclave. Separately autoclave cysteine HCl and Na2S x 9H2O as 5% solutions under N2. Before inoculation, aseptically and anaerobically add the sterile cysteine and sulfide solutions, then pressurize the inoculated bottles to 200 kPa H2-CO2 (80:20, v/v).
Prepare clarified rumen fluid by preheating rumen content at 120 C for 15 minutes and using the supernatant after centrifugation at 25,000 x g for 15 minutes.
Use JCM Medium 266 without its fatty acid mixture.

Solutions

Rumen fluid, clarified (JCM Medium 266)

Dose: 300.0 ML_PER_L

JCM Medium 773 adds 300.0 ml/L clarified rumen fluid from JCM Medium 266.

Trace minerals (JCM Medium 151)

Dose: 10.0 ML_PER_L

JCM Medium 773 adds 10.0 ml/L Trace minerals from JCM Medium 151.

Trace vitamins (JCM Medium 197)

Dose: 10.0 ML_PER_L

JCM Medium 773 adds 10.0 ml/L Trace vitamins from JCM Medium 197.

Target organisms

No target organisms are recorded.

Provenance

Timestamp	Curator	Action	Notes
2026-01-27T06:35:55.744307Z	togo-import	Imported from TOGO Medium	Source: JCM, ID: M802
2026-01-28T19:48:08.669010Z	chebi-enrichment	Added CHEBI ontology terms to ingredients	Enriched using MicrobeMediaParam and MediaDive chemical mappings
2026-02-03T00:00:00+00:00	schema-defaulter-v1.0	Applied schema defaults and normalizations	Added default concentration for ingredient: Carbon dioxide gas; Added default concentration for ingredient: N2; Added default concentration for ingredient: Hydrogen gas
2026-03-09T08:04:13.982910+00:00	solution-migrator-v1.0	Migrated solutions from ingredients to solutions array	Moved 3 solution(s) from ingredients
2026-03-13T06:20:12.164899Z	culturemech-id-assigner-v1.0	Assigned CultureMech ID	Assigned stable identifier: CultureMech:010214
2026-03-14T00:00:00+00:00	schema-defaulter-v1.0	Applied schema defaults and normalizations	Added default solution name; Added default solution name; Added default solution name
2026-03-17T06:29:43.000000Z	mediaingredientmech-enrichment-v1.0	Added MediaIngredientMech links	Linked ingredients to MediaIngredientMech identifiers
2026-03-17T19:46:31.106401+00:00	normalize_media_names_to_snake_case.py	NAME_NORMALIZED	Normalized name from "Rumen Fluid Medium" to "rumen_fluid_medium" for consistent matching
2026-05-16T09:03:21.005052+00:00	migrate_legacy_fields.py	MIGRATED_LEGACY_FIELDS	curation_history.date->timestamp=2
2026-06-05T07:40:44.590023+00:00	mim-legacy-to-chebi-migration-v1.0	Refreshed legacy MediaIngredientMech links to CHEBI keying	Replaced 7 legacy mediaingredientmech_term link(s) with mediaingredientmech_chebi_term (id-safe: ingredient's own CHEBI + label). MIM deprecated the MediaIngredientMech:NNNNNN id scheme.
2026-06-10T06:57:33.124485+00:00	mgso4-heptahydrate-term-fix-v1.0	Corrected MgSO4·7H2O primary term grounding to heptahydrate	Re-grounded 1 heptahydrate-labelled ingredient(s) from CHEBI:32599 (generic magnesium sulfate) to CHEBI:31795 (magnesium sulfate heptahydrate). Generic/hexahydrate labels untouched.
2026-06-10T07:46:51.684521+00:00	primary-term-label-canonicalization-v1.0	Canonicalized primary term label to OBO canonical (OAK)	3 primary term(s); ingredient name resolved by OAK sqlite:obo:chebi.
2026-09-12T00:00:00-07:00	repair_jcm_j773_togo_m802_rumen_score15.py	RESOLVED_JCM_J773_TOGO_M802_RUMEN_SCORE15	Repaired the JCM Medium 773 formula from TOGO M802, corrected TOGO g/L import artifacts for water, trace milligram salts, and stock additions, grounded all direct defined chemicals and gases, added the pH 6.5 instruction, and preserved the clarified rumen fluid, trace-mineral, and trace-vitamin external boundaries.