Medium for Sulfate Reducers (ATCC Medium 1282)

[所属分类:培养基配方] [发布时间:2021-5-20] [发布人:网站管理员2] [阅读次数:] [返回]

Medium for Sulfate Reducers
(ATCC Medium 1282)

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per 1050.0mL:

Modified Baar’s medium
for sulfate reducers........................................................1020.0mL
Organic acid solution...............................................................10.0mL
Wolfe’s vitamin solution..........................................................10.0mL
Wolfe’s mineral solution..........................................................10.0mL
pH 7.5 ± 0.2 at 25°C
Modified Baar’s Medium for Sulfate Reducers:
Composition per 1020.0mL:
Component I ..........................................................................400.0mL
Component III........................................................................400.0mL
Component II.........................................................................200.0mL
Fe(NH 4 ) 2 (SO 4 ) 2 (5% solution).................................................20.0mL
Component I:
Composition per 400.0mL:
Sodium citrate...............................................................................5.0g
MgSO 4 ..........................................................................................2.0g
CaSO 4 ...........................................................................................1.0g
NH 4 Cl ...........................................................................................1.0g
Preparation of Component I: Add components to distilled/deion-
ized water and bring volume to 400.0mL. Mix thoroughly. Adjust pH
to 7.5. Autoclave for 15 min at 15 psi pressure–121°C.
Component II:
Composition per 200.0mL:
K 2 HPO 4 .........................................................................................0.5g
Preparation of Component II: Add K 2 HPO 4 to distilled/deion-
ized water and bring volume to 200.0mL. Mix thoroughly. Adjust pH
to 7.5. Autoclave for 15 min at 15 psi pressure–121°C.
Component III:
Composition per 400.0mL:
Sodium lactate...............................................................................3.5g
Yeast extract..................................................................................1.0g
Preparation of Component III: Add components to distilled/de-
ionized water and bring volume to 400.0mL. Mix thoroughly. Adjust
pH to 7.5. Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Modified Baar’s Medium for Sulfate Reduc-
ers: Aseptically combine the three sterile solutions, except the
Fe(NH 4 ) 2 (SO 4 ) 2 solution. Mix thoroughly. Distribute 5.0mL volumes
into tubes under 97% N 2 + 3% H 2 . Add medium to tubes while still
warm to exclude as much O 2 as possible. Prepare a 5% solution of fer-
rous ammonium sulfate, Fe(NH 4 ) 2 (SO 4 ) 2 . Sterilize by filtration. Add
0.2mL of sterile Fe(NH 4 ) 2 (SO 4 ) 2 solution to 10.0mL of medium imme-
diately prior to inoculation.
Organic Acid Solution:
Composition per 100.0mL:
Butyric acid..............................................................................5.18mL
Caproic acid...............................................................................2.4mL
Octanoic acid ...........................................................................1.25mL
Preparation of Organic Acid Solution: Add components to dis-
tilled/deionized water and bring volume to 75.0mL. Adjust pH to 7.0
with 5N NaOH. Bring volume to 100.0mL with distilled/deionized wa-
ter. Filter sterilize.
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
Thiamine·HCl ............................................................................5.0mg
Riboflavin ..................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
Calcium pantothenate ................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Thioctic acid ..............................................................................5.0mg
Biotin .........................................................................................2.0mg
Folic acid....................................................................................2.0mg
Cyanocobalamin .....................................................................100.0μg
Preparation of Wolfe’s Vitamin Solution: Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Filter sterilize.
Wolfe’s Mineral Solution:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·H 2 O..................................................................................0.5g
FeSO 4 ·7H 2 O..................................................................................0.1g
CoCl 2 ·6H 2 O..................................................................................0.1g
CaCl 2 .............................................................................................0.1g
ZnSO 4 ·7H 2 O.................................................................................0.1g
CuSO 4 ·5H 2 O...............................................................................0.01g
AlK(SO 4 ) 2 ·12H 2 O.......................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O..........................................................................0.01g
Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 6.5 with KOH. Add remaining components. Add distilled/deionized
water to 1.0L. Filter sterilize.
Preparation of Medium:  To each test tube containing 10.0mL of
modified Baar’s medium for sulfate reducers, aseptically add 0.1mL of
sterile organic acid solution, 0.1mL of sterile Wolfe’s vitamin solution,
and 0.1mL of sterile Wolfe’s mineral solution immediately prior to in-
oculation.
Use:  For the cultivation and maintenance of Desulfotomaculum ther-
mobenzoicum and Desulfovibrio sapovorans.