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DSMZ

DSMZ 1494 . . OMIZ-PAT Medium (modified)

This medium was extracted automatically and has not been curated by experts, yet.

Culture medium recipe

Vol.
×
Medium for Treponema medium subsp. bovis DSM 18689

This strain uses the standard medium without modifications.

Growth conditions: 7-14 days anaerobic 37 °C pH 7.00

Main sol. 1494 #

ml
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Distilled water 100 ml - -
Solution A 10 ml - -
Solution B 10 ml - -
Solution C 10 ml - -
Solution D 10 ml - -
Solution E 10 ml - -
Solution F 10 ml - -
Solution G 10 ml - -
Solution H 200 µl - -
Solution I 8 µl - -
Solution J 2 µl - -
Solution K 40 µl - -
Solution L 2 ml - -
Trace elements I
(pre-dilute 1:1000; use) 		2 ml - -
Trace elements II
(pre-dilute 1:1000; use) 		2 ml - -
Trace elements III
(pre-dilute 1:1000; use) 		200 µl - -
Distilled water
(add up to 200 ml) 		200 ml - -
Supplement 1 200 µl - -
Supplement 2 2 ml - -
Supplement 3 2 ml - -
Supplement 4 200 µl - -
Supplement 5 2 ml - -
1 Add 10 ml rabbit serum, flush the medium with N2CO2, adjust the pH to 6.9 and filter-sterilize the completed medium into tubes.
Rabbit serum 10 ml - -
2 This recipe describes the preparation of 200 ml medium, which can be used for about 4 weeks when kept at 4°C.

Solution A #

100
Compound Amount Unit Conc. [g/L] Conc. [mM]
L-Alanine 90 mg 0.9 10.102
L-Arginine 350 mg 3.5 20.092
L-Asparagine 300 mg 3 22.707
L-Aspartic acid 236 mg 2.36 17.731
L-Cysteine HCl 700 mg 7 44.411
L-Glutamine 1360 mg 13.6 93.061
L-Glutamic acid 600 mg 6 40.78
Glycine 150 mg 1.5 19.981
L-Histidine 1240 mg 12.4 79.923
L-Isoleucine 262 mg 2.62 19.974
L-Leucine 262 mg 2.62 19.974
L-Lysine HCl 364 mg 3.64 -
L-Methionine 300 mg 3 20.106
L-Ornithine HCl 340 mg 3.4 -
L-Phenylalanine 330 mg 3.3 19.977
L-Proline 230 mg 2.3 19.977
L-Serine 1050 mg 10.5 99.914
L-Threonine 240 mg 2.4 20.148
L-Tryptophan 204 mg 2.04 9.989
L-Tyrosine 180 mg 1.8 9.934
L-Valine 234 mg 2.34 19.974
Distilled water 100 ml - -
1 Prepare Solutions A, flush with nitrogen to make them anoxic, filter-sterilize and store at -20°C

Solution B #

100
Compound Amount Unit Conc. [g/L] Conc. [mM]
KCl 1936 mg 19.36 259.691
MgSO4 x 7 H2O 500 mg 5 20.286
NaH2PO4 x H2O 280 mg 2.8 23.338
NH4Cl 3200 mg 32 598.243
Citric acid , trisodium salt 400 mg 4 20.82
Distilled water 100 ml - -
1 Prepare Solutions B, flush with nitrogen to make them anoxic, filter-sterilize and store at -20°C

Solution C #

100
Compound Amount Unit Conc. [g/L] Conc. [mM]
Calcium D-(+)-pantothenate 10 mg 0.1 -
Choline chloride 100 mg 1 7.162
Myo-inositol 100 mg 1 5.551
Thiamine HCl 10 mg 0.1 0.296
Thiamin pyrophosphate 10 mg 0.1 0.236
Coenzym A , sodium salt 2 mg 0.02 0.026
Flavin adenine dinucleotide
(FAD) 		2 mg 0.02 0.025
ß-nicotinamide adenine dinucleotide
(NAD) 		2 mg 0.02 -
2-Mercaptoethanesulfonic acid , sodium salt
(MES) 		20 mg 0.2 1.406
N-acetylmuramic acid 50 mg 0.5 1.705
N-Acetylglucosamine 400 mg 4 18.082
D-glucuronic acid , sodium salt 1600 mg 16 82.415
D-galacturonic acid 1600 mg 16 82.415
Pyruvic acid , sodium salt 1100 mg 11 124.915
Fumaric acid , disodium salt 1000 mg 10 86.155
Formic acid , sodium salt 600 mg 6 130.363
Distilled water 100 ml - -
1 Prepare Solutions C, flush with nitrogen to make them anoxic, filter-sterilize and store at -20°C

Solution D #

100
Compound Amount Unit Conc. [g/L] Conc. [mM]
Pyridoxal hydrochloride 10.0 mg 0.1 0.491
Pyridoxal phosphate 10.0 mg 0.1 0.405
D , L-lactic acid, sodium salt 1120.0 mg 11.2 97.318
Hypoxanthine 2.8 mg 0.028 0.206
Uracil 2.2 mg 0.022 0.196
Thymidine 4.8 mg 0.048 0.198
N-(2-acetamido)-2-aminoethanesulfonic acid 3.6 mg 0.036 0.198
DL-carnitine 400.0 mg 4 24.814
Putrescine 2 HCl 10.0 mg 0.1 -
Phenol red , sodium salt 20.0 mg 0.2 0.564
Distilled water 100.0 ml - -
1 Prepare Solutions D, flush with nitrogen to make them anoxic, filter-sterilize and store at -20°C

Solution E #

100
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
D-Glucose 4.0 g 40 222.025
D-Fructose 1.6 g 16 88.81
D-Maltose 1.6 g 16 46.743
D-Mannitol 1.6 g 16 87.83
D-Mannose 4.0 g 40 222.03
D-Arabinose 4.0 g 40 266.436
L-Fucose 4.0 g 40 243.665
D-Trehalose 4.0 g 40 116.858
D-Sucrose 4.0 g 40 116.857
L-Rhamnose 4.0 g 40 243.665
Distilled water 100.0 ml - -
1 Prepare Solutions E, flush with nitrogen to make them anoxic, filter-sterilize and store at -20°C

Solution F #

100
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
CaCl2 x H2O 300 mg 3 27.031
Distilled water 100 ml - -
1 Prepare Solution F, flush with nitrogen to make it anoxic, filter-sterilize and store at 4°C

Solution G #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
Asialofetuin 1 mg/ml - -
1 Prepare Solutions G, flush with nitrogen to make them anoxic, filter-sterilize and store at -20°C

Solution H #

100
Compound Amount Unit Conc. [g/L] Conc. [mM]
Vitamine B12 5 mg 0.05 0.037
D-(+)-biotin 5 mg 0.05 0.205
Folic acid 5 mg 0.05 0.113
Folinic acid , calcium salt 10 mg 0.1 0.211
Nicotinamide 5 mg 0.05 0.409
Nicotinic acid 10 mg 0.1 0.812
Riboflavine 1 mg 0.01 0.027
Distilled water 100 ml - -
1 Prepare Solutions H, flush each with nitrogen to make them anoxic, filter-sterilize and store at -20°C

Solution I #

Compound Amount Unit Conc. [g/L] Conc. [mM]
Isobutyric acid 10 µl - -
2-Methylbutyric acid 10 µl - -
Valeric acid 10 µl - -
Isovaleric acid 10 µl - -
1 Prepare Solution I and store at -20°C

Solution J #

1
Compound Amount Unit Conc. [g/L] Conc. [mM]
DL-alpha-lipoic acid 1.0 mg/ml - -
2-Mercaptoethanol 0.1 ml - -
Ethanol 0.9 ml - -
1 D,L-alpha-lipoic acid (1 mg/ml); in 2-mercaptoethanol-ethanol (1:10); store at -20°C

Solution K #

1
Compound Amount Unit Conc. [g/L] Conc. [mM]
Hemin 0.2 mM 0.13 0.2
NaOH 10.0 mM 0.4 10
1 Hemin (0.2 mM) in NaOH (10 mM)

Solution L #

140
Equipment needed: Filter Centrifuge
Compound Amount Unit Conc. [g/L] Conc. [mM]
Yeast extract
(fresh) 		42 g 300 -
Distilled water 140 ml - -
1 Homogenize 42 g fresh baker yeast in 140 ml distilled water, incubate for 24h at 56°C and centrifuge to harvest the supernatant. Adjust pH to 7 with NaOH, make anoxic, filter-sterilize and store at -80°C

Trace elements I #

100
Compound Amount Unit Conc. [g/L] Conc. [mM]
CuSO4 7.89 mg 0.079 0.494
MnSO4 x H2O 169.00 mg 1.69 9.999
ZnSO4 x 7 H2O 2.87 g 28.7 99.805
HCl
(10 mM) 		100.00 ml 0.036 1
1 Prepare Trace elements I, flush with nitrogen to make them anoxic, filter-sterilize and store at 4°C

Trace elements II #

100
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2SeO3 173 mg 1.73 10.004
HCl
(10mM) 		100 ml 0.036 1
1 Prepare Trace elements II, flush with nitrogen to make them anoxic, filter-sterilize and store at 4°C

Trace elements III #

100
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
NiSO4 x 6 H2O 13.1 mg 0.131 0.846
SnCl2 x 2 H2O 11.8 mg 0.118 0.523
NaVO3 61.0 mg 0.61 5.003
(NH4)6Mo7O24 x 4 H2O 1240.0 mg 12.4 -
HCl
(10 mM) 		100.0 ml 0.036 1
1 Prepare Trace elements III, flush with nitrogen to make them anoxic, filter-sterilize and store at 4°C

Supplement 1 #

1
Compound Amount Unit Conc. [g/L] Conc. [mM]
Fe SO4 x 7 H2O 10 mM 2.78 10
HCl 10 mM 0.365 10
1 FeSO4 x 7H2O (10 mM) in HCl (10 mM)
2 Prepare Supplements 1, flush with nitrogen to make them anoxic, filter-sterilize and store at -20°C

Supplement 2 #

2
Compound Amount Unit Conc. [g/L] Conc. [mM]
Ascorbic acid 10 %(w/v) 100 567.795
Distilled water 2 ml - -
1 Ascorbic acid (10%; w/v); in distilled water, neutralize with NaOH
2 Prepare Supplements 2, flush with nitrogen to make them anoxic, filter-sterilize and store at -20°C

Supplement 3 #

2
Compound Amount Unit Conc. [g/L] Conc. [mM]
Glutathione 0.5 M 153.66 500
Distilled water 2.0 ml - -
1 Glutathion (0.5 M); in distilled water, neutralize with NaOH
2 Prepare Supplements 3, flush with nitrogen to make them anoxic, filter-sterilize and store at -20°C

Supplement 4 #

1
Compound Amount Unit Conc. [g/L] Conc. [mM]
Cholesterol 1 mg/ml - -
Ethanol
(100 %) 		1 ml 1 21.706
1 Cholesterol (1 mg/ml); in Ethanol (100%); store at -20°C

Supplement 5 #

2
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaHCO3 10 %(w/v) 100 1190.38
Distilled water 2 ml - -
1 NaHCO3 (10%; w/v) in distilled water, store at 4°C

Metadata

Last modified: 24.02.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 6.9
Equipment needed: Centrifuge, Filter
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Associated strains

All strains for this medium: 17
Strains with modifications: 3
Treponema medium subsp. bovis DSM 18689

No modifications necessary for this strain.

Molecular composition

Compound Conc. [g/L] Conc. [mM]
(NH4)6Mo7O24 x 4 H2O 12.4 -
Yeast extract 3 -
Fe SO4 x 7 H2O 2.78 10
D-Arabinose 2 13.322
L-Rhamnose 2 12.183
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Cultivation metadata from 17 strains BacDive

phyla 4 17 strains
oxygen anaerobe 9 strains
temperature mesophilic 13 strains