Preparation of Medium: Add components, except CaCO3, Na2CO3 solution, and vitamin K1 solution, to distilled/deionized water and bring volume to 990.0mL.. Preparation of Medium: Add compo
Trang 1Thioglycolate Medium, USP 1775
Thioglycolate Medium without Indicator
Compositionper liter:
Pancreatic digest of casein 17.0g
Glucose 6.0g
Papaic digest of soybean meal 3.0g
NaCl 2.5g
Agar 0.7g
Sodium thioglycolate 0.5g
L-Cystine 0.25g
Na2SO3 0.1g
pH 7.0 ± 0.2 at 25°C
Source: This medium is available as a premixed powder from Oxoid
Unipath
Preparation of Medium: Add components to distilled/deionized
water and bring volume to 1.0L Mix thoroughly Distribute into tubes
or flasks Autoclave for 15 min at 15 psi pressure–121°C Prepare
freshly or boil and cool the medium just before use
Use: For the growth of aerobic and anaerobic microorganisms in
diag-nostic bacteriology
Thioglycolate Medium without Indicator-135C
Compositionper liter:
Pancreatic digest of casein 17.0g
Glucose 6.0g
Papaic digest of soybean meal 3.0g
NaCl 2.5g
Agar 0.7g
Sodium thioglycolate 0.5g
Na2SO3 0.1g
L-Cystine 0.25g
pH 7.0 ± 0.2 at 25°C
Source: This medium is available as a premixed powder from BD
Di-agnostic Systems
Preparation of Medium: Add components to distilled/deionized
water and bring volume to 1.0L Mix thoroughly Gently heat while
stirring and bring to boiling Distribute into tubes or flasks, filling them
half full For maintenance of cultures, a small quantity of CaCO3 may
be added to tubes before adding medium Autoclave for 15 min at 13
psi pressure–118°C Prepare freshly or boil and cool the medium just
before use Store prepared medium at 2°–8°C in the dark
Use: For the isolation and cultivation of a wide variety of
microorgan-isms, particularly obligate anaerobes, from clinical specimens and
other materials
Thioglycolate Medium without Indicator with Hemin
(Thioglycolate Medium, Supplemented)
Compositionper liter:
Pancreatic digest of casein 17.0g
CaCO3, chips or powder 10.0g
Glucose 6.0g
Papaic digest of soybean meal 3.0g
NaCl 2.5g
Agar 0.7g
Sodium thioglycolate 0.5g
L-Cystine 0.25g
Na2SO3 0.1g
Hemin 5.0mg
Na2CO3 solution 10.0mL Vitamin K1 solution 10.0mL
pH 7.2 ± 0.2 at 25°C
Na 2 CO 3 Solution:
Compositionper 10.0mL:
Na2CO3 1.0g
Preparation of Na 2 CO 3 Solution: Add Na2CO3 to distilled/de-ionized water and bring volume to 10.0mL Mix thoroughly Filter ster-ilize
Vitamin K 1 Solution:
Compositionper 100.0mL:
Vitamin K1 1.0g Ethanol, absolute 99.0mL
Preparation of Vitamin K 1 Solution: Add vitamin K1 to 99.0mL
of absolute ethanol Mix thoroughly
Preparation of Medium: Add components, except CaCO3, Na2CO3 solution, and vitamin K1 solution, to distilled/deionized water and bring volume to 990.0mL Mix thoroughly Gently heat and bring to boiling Add 0.1g of CaCO3 chips or powder to each of 100 test tubes Distribute broth into the same tubes in 10.0mL volumes Autoclave for
15 min at 15 psi pressure–121°C Cool to 45°–50°C Aseptically add 0.1mL of sterile Na2CO3 solution and 0.1mL of sterile vitamin K1 so-lution to each tube Mix thoroughly
Use: For the cultivation of a wide variety of obligate anaerobes
Thioglycolate Medium, Supplemented
See: Thioglycolate Medium without Indicator with Hemin
Thioglycolate Medium, USP
Compositionper liter:
Pancreatic digest of casein 15.0g Glucose 5.5g Yeast extract 5.0g NaCl 2.5g Agar 0.5g
L-cystine 0.5g Sodium thioglycolate 0.5g Resazurin 1.0mg
pH 7.1 ± 0.2 at 25°C
Source: This medium is available as a premixed powder from Oxoid Unipath
Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Gently heat and bring
to boiling Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C
Use: For the cultivation of both aerobic and anaerobic organisms in the performance of sterility tests
Thioglycolate Medium with Vitamin K 1 and Hemin
See: Thioglycolate Medium, Enriched Thioglycolate Peptone Glucose Yeast Extract Medium
Thioglycolate Potato Liver Medium
Trang 21776 Thiol Broth
Thiol Broth
Composition per liter:
Proteose peptone, No 3 10.0g
Thiol complex 8.0g
Yeast extract 5.0g
NaCl 5.0g
Glucose 1.0g
p-Aminobenzoic acid 0.05g
pH 7.1 ± 0.2 at 25°C
Source: This medium is available as a premixed powder from BD
Di-agnostic Systems
Preparation of Medium: Add components to distilled/deionized
water and bring volume to 1.0L Mix thoroughly Gently heat and bring
to boiling Distribute into tubes or flasks For neutralization of
penicil-lin, distribute medium into tubes to a depth of 60.0mm For
neutraliza-tion of streptomycin, distribute medium into tubes in shallow layers
Autoclave for 15 min at 15 psi pressure–121°C
Use: For the cultivation of bacteria from body fluids and other
mate-rials containing penicillin, streptomycin, or sulfonamides Also used
for the cultivation and maintenance of Bifidobacterium species.
Thiohalophilus Medium
(DSMZ Medium 1058)
Composition per liter:
NaCl 120.0g
K2HPO4 1.5g
NH4Cl 0.5g
Bicarbonate solution 10.0mL
Calcium chloride solution 10.0mL
Magnesium chloride solution 10.0mL
Seven vitamin solution 1.0mL
Trace elements solution SL-10 with EDTA 1.0mL
Carbonate solution variable
pH 7.6 ± 0.2 at 25°C
Thiosulfate Solution:
Composition per10.0mL:
Na2S2O3·5H2O 5.0g
Preparation of Thiosulfate Solution: Add components to
dis-tilled/deionized water and bring volume to 10.0mL Mix thoroughly
Sparge with 100% N2 Filter sterilize
Bicarbonate Solution :
Compositionper 10.0mL:
NaHCO3 5.0g
Preparation of Bicarbonate Solution: Add NaHCO3 to distilled/
deionized water and bring volume to 10.0mL Mix thoroughly Sparge
with a gas mixture of 80% N2 + 20% CO2 Filter sterilize
Carbonate Solution :
Compositionper 10.0mL:
Na2CO3 2.0g
Preparation of Carbonate Solution: Add Na2CO3 to
distilled/de-ionized water and bring volume to 10.0mL Mix thoroughly Sparge
with a gas mixture of 80% N2 + 20% CO2 Filter sterilize
Magnesium Chloride Solution:
Compositionper 10.0mL:
MgCl2·6H2O 0.4g
Preparation of Magnesium Chloride Solution: Add 0.4g of
MgCl2·6H2O to distilled/deionized water and bring volume to 10.0mL
Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Cool
to room temperature Sparge with 100% N2
Calcium Chloride Solution:
Compositionper 10.0mL:
CaCl2·2H2O 0.05g
Preparation of Calcium Chloride Solution: Add components to distilled/deionized water and bring volume to 10.0mL Mix
thorough-ly Autoclave for 15 min at 15 psi pressure–121°C Cool to room tem-perature Sparge with 100% N2 gas
Seven Vitamin Solution:
Compositionper liter:
Pyridoxine·HCl 300.0mg Nicotinic acid 200.0mg Thiamine·HCl 200.0mg Calcium DL-pantothenate 100.0mg Cyanocobalamine 100.0mg
p-Aminobenzoic acid 80.0mg
D(+)-Biotin 20.0mg
Preparation of Seven Vitamin Solution: Add components to dis-tilled/deionized water and bring volume to 1.0L Mix thoroughly Filter sterilize Sparge with 100% N2
Trace Elements Solution SL-10 with EDTA:
Compositionper liter:
FeCl2·4H2O 1.5g
Na2-EDTA 0.5g CoCl2·6H2O 190.0mg MnCl2·4H2O 100.0mg ZnCl2 70.0mg
Na2MoO4·2H2O 36.0mg NiCl2·6H2O 24.0mg
H3BO3 6.0mg CuCl2·2H2O 2.0mg HCl (25% solution) 10.0mL
Preparation of Trace Elements Solution SL-10 with EDTA:
Add FeCl2·4H2O to 10.0mL of HCl solution Mix thoroughly Add dis-tilled/deionized water and bring volume to 1.0L Add remaining com-ponents Mix thoroughly Sparge with 100% N2 Autoclave for 15 min
at 15 psi pressure–121°C Adjust pH to 7.0
Preparation of Medium: Add components, except magnesium chloride, thiosulfate, bicarbonate, and vitamin solutions, to distilled/ deionized water and bring volume to 970.0mL Mix thoroughly Sparge with 80% N2 + 20% CO2 gas mixture for at least 30 min to remove dis-solved oxygen and to saturate the solution with CO2 Dispense into an-aerobic culture vessels (e.g., Balch tubes) to 1/2 volume under air atmosphere Close vials with butyl rubber septa to prevent free ex-change of oxygen with the external atmosphere Autoclave for 15 min
at 15 psi pressure–121°C Cool to room temperature Aseptically add magnesium chloride, thiosulfate, bicarbonate, and vitamin solutions Mix thoroughly Adjust pH to 7.5–7.8 using a sterile solution of sodium carbonate (5% w/v)
Use: For the cultivation of Thiohalomonas denitrificans,
Thiohalo-philus thiocyanatoxydans, and Thiomicrospira halophila
Thiohalophilus Medium
(DSMZ Medium 1058)
Composition per liter:
NaCl 233.0g
K2HPO4 1.5g
Trang 3Thiohalophilus Medium 1777
NH4Cl 0.5g
Bicarbonate solution 10.0mL
Calcium chloride solution 10.0mL
Magnesium chloride solution 10.0mL
Seven vitamins solution 1.0mL
Trace elements solution SL-10 with EDTA 1.0mL
Carbonate solution variable
pH 7.2 ± 0.2 at 25°C
Thiosulfate Solution:
Composition per10.0mL:
Na2S2O3·5H2O 5.0g
Preparation of Thiosulfate Solution: Add Na2S2O3·5H2O to
dis-tilled/deionized water and bring volume to 10.0mL Mix thoroughly
Sparge with 100% N2 Filter sterilize
Bicarbonate Solution :
Compositionper 10.0mL:
NaHCO3 5.0g
Preparation of Bicarbonate Solution: Add NaHCO3 to distilled/
deionized water and bring volume to 10.0mL Mix thoroughly Sparge
with a gas mixture of 80% N2 + 20% CO2 Filter sterilize
Carbonate Solution :
Compositionper 10.0mL:
Na2CO3 2.0g
Preparation of Carbonate Solution: Add Na2CO3 to
distilled/de-ionized water and bring volume to 10.0mL Mix thoroughly Sparge
with a gas mixture of 80% N2 + 20% CO2 Filter sterilize
Magnesium Chloride Solution:
Compositionper 10.0mL:
MgCl2·6H2O 0.4g
Preparation of Magnesium Chloride Solution: Add 0.4g of
MgCl2·6H2O to distilled/deionized water and bring volume to 10.0mL
Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Cool
to room temperature Sparge with 100% N2
Calcium Chloride Solution:
Compositionper 10.0mL:
CaCl2·2H2O 0.05g
Preparation of Calcium Chloride Solution: Add CaCl2·2H2O
to distilled/deionized water and bring volume to 10.0mL Mix
thor-oughly Autoclave for 15 min at 15 psi pressure–121°C Cool to room
temperature Sparge with 100% N2
Seven Vitamin Solution:
Compositionper liter:
Pyridoxine·HCl 300.0mg
Nicotinic acid 200.0mg
Thiamine·HCl 200.0mg
Calcium DL-pantothenate 100.0mg
Cyanocobalamine 100.0mg
p-Aminobenzoic acid 80.0mg
D(+)-Biotin 20.0mg
Preparation of Seven Vitamin Solution: Add components to
dis-tilled/deionized water and bring volume to 1.0L Mix thoroughly Filter
sterilize Sparge with 100% N2
Trace Elements Solution SL-10 with EDTA:
Compositionper liter:
FeCl2·4H2O 1.5g
Na2-EDTA 0.5g
CoCl2·6H2O 190.0mg
MnCl2·4H2O 100.0mg ZnCl2 70.0mg
Na2MoO4·2H2O 36.0mg NiCl2·6H2O 24.0mg
H3BO3 6.0mg CuCl2·2H2O 2.0mg HCl (25% solution) 10.0mL
Preparation of Trace Elements Solution SL-10 with EDTA:
Add FeCl2·4H2O to 10.0mL of HCl solution Mix thoroughly Add dis-tilled/deionized water and bring volume to 1.0L Add remaining com-ponents Mix thoroughly Sparge with 100% N2 Autoclave for 15 min
at 15 psi pressure–121°C Adjust pH to 7.0
Preparation of Medium: Add components, except magnesium chloride, thiosulfate, bicarbonate, and vitamin solutions, to distilled/ deionized water and bring volume to 970.0mL Mix thoroughly Sparge with 80% N2 + 20% CO2 gas mixture for at least 30 min to remove dis-solved oxygen and to saturate the solution with CO2 Dispense into an-aerobic culture vessels (e.g., Balch tubes) to 1/2 volume under air atmosphere Close vials with butyl rubber septa to prevent free ex-change of oxygen with the external atmosphere Autoclave for 15 min
at 15 psi pressure–121°C Cool to room temperature Aseptically add magnesium chloride, thiosulfate, bicarbonate, and vitamin solutions Mix thoroughly Adjust pH to 7.2 using a sterile solution of sodium car-bonate (5% w/v)
Use: For the cultivation of Thiomicrospira halophila DSM 15071
Thiohalophilus Medium
(DSMZ Medium 1058)
Composition per liter:
NaCl 180.0g
K2HPO4 1.5g KNO3 1.0g
NH4Cl 0.5g Bicarbonate solution 10.0mL Calcium chloride solution 10.0mL Magnesium chloride solution 10.0mL Seven vitamin solution 1.0mL Trace elements solution SL-10 with EDTA 1.0mL Carbonate solution Variable
pH 7.6 ± 0.2 at 25°C
Thiosulfate Solution:
Composition per10.0mL:
Na2S2O3·5H2O 5.0g
Preparation of Thiosulfate Solution: Add Na2S2O3·5H2O to dis-tilled/deionized water and bring volume to 10.0mL Mix thoroughly Sparge with 100% N2 Filter sterilize
Bicarbonate Solution :
Compositionper 10.0mL:
NaHCO3 5.0g
Preparation of Bicarbonate Solution: Add NaHCO3 to distilled/ deionized water and bring volume to 10.0mL Mix thoroughly Sparge with a gas mixture of 80% N2 + 20% CO2 Filter sterilize
Carbonate Solution :
Compositionper 10.0mL:
Na2CO3 2.0g
Preparation of Carbonate Solution: Add Na2CO3 to distilled/de-ionized water and bring volume to 10.0mL Mix thoroughly Sparge with a gas mixture of 80% N2 + 20% CO2 Filter sterilize
Trang 41778 Thiohalophilus Medium
Magnesium Chloride Solution:
Compositionper 10.0mL:
MgCl2·6H2O 0.4g
Preparation of Magnesium Chloride Solution: Add 0.4g of
MgCl2·6H2O to distilled/deionized water and bring volume to 10.0mL
Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Cool
to room temperature Sparge with 100% N2
Calcium Chloride Solution:
Compositionper 10.0mL:
CaCl2·2H2O 0.05g
Preparation of Calcium Chloride Solution: Add CaCl2·2H2O
to distilled/deionized water and bring volume to 10.0mL Mix
thor-oughly Autoclave for 15 min at 15 psi pressure–121°C Cool to room
temperature Sparge with 100% N2
Seven Vitamin Solution:
Compositionper liter:
Pyridoxine·HCl 300.0mg
Nicotinic acid 200.0mg
Thiamine·HCl 200.0mg
Calcium DL-pantothenate 100.0mg
Cyanocobalamine 100.0mg
p-Aminobenzoic acid 80.0mg
D(+)-Biotin 20.0mg
Preparation of Seven Vitamin Solution: Add components to
dis-tilled/deionized water and bring volume to 1.0L Mix thoroughly Filter
sterilize Sparge with 100% N2
Trace Elements Solution SL-10 with EDTA:
Compositionper liter:
FeCl2·4H2O 1.5g
Na2-EDTA 0.5g
CoCl2·6H2O 190.0mg
MnCl2·4H2O 100.0mg
ZnCl2 70.0mg
Na2MoO4·2H2O 36.0mg
NiCl2·6H2O 24.0mg
H3BO3 6.0mg
CuCl2·2H2O 2.0mg
HCl (25% solution) 10.0mL
Preparation of Trace Elements Solution SL-10 with EDTA:
Add FeCl2·4H2O to 10.0mL of HCl solution Mix thoroughly Add
dis-tilled/deionized water and bring volume to 1.0L Add remaining
com-ponents Mix thoroughly Sparge with 100% N2 Autoclave for 15 min
at 15 psi pressure–121°C Adjust pH to 7.0
Preparation of Medium: Add components, except magnesium
chloride, thiosulfate, bicarbonate, and vitamin solutions, to distilled/
deionized water and bring volume to 970.0mL Mix thoroughly Sparge
with 80% N2 + 20% CO2 gas mixture for at least 30 min to remove
dis-solved oxygen and to saturate the solution with CO2 Dispense into
an-aerobic culture vessels (e.g., Balch tubes) to 1/2 volume under air
atmosphere Close vials with butyl rubber septa to prevent free
ex-change of oxygen with the external atmosphere Autoclave for 15 min
at 15 psi pressure–121°C Cool to room temperature Aseptically add
magnesium chloride, thiosulfate, bicarbonate, and vitamin solutions
Mix thoroughly Adjust pH to 7.5–7.8 using a sterile solution of sodium
carbonate (5% w/v)
Use: For the cultivation of Thiohalomonas denitrificans.
Thiohalophilus Medium
(DSMZ Medium 1058)
Composition per liter:
NaCl 180.0g
K2HPO4 1.5g KNO3 2.0g
NH4Cl 0.5g Bicarbonate solution 10.0mL Calcium chloride solution 10.0mL Magnesium chloride solution 10.0mL Seven vitamin solution 1.0mL Trace elements solution SL-10 with EDTA 1.0mL Carbonate solution Variable
pH 7.6 ± 0.2 at 25°C
Thiosulfate Solution:
Composition per10.0mL:
Na2S2O3·5H2O 5.0g
Preparation of Thiosulfate Solution: Add Na2S2O3·5H2O to dis-tilled/deionized water and bring volume to 10.0mL Mix thoroughly Sparge with 100% N2 Filter sterilize
Bicarbonate Solution :
Compositionper 10.0mL:
NaHCO3 5.0g
Preparation of Bicarbonate Solution: Add NaHCO3 to distilled/ deionized water and bring volume to 10.0mL Mix thoroughly Sparge with a gas mixture of 80% N2 + 20% CO2 Filter sterilize
Carbonate Solution :
Compositionper 10.0mL:
Na2CO3 2.0g
Preparation of Carbonate Solution: Add Na2CO3 to distilled/de-ionized water and bring volume to 10.0mL Mix thoroughly Sparge with a gas mixture of 80% N2 + 20% CO2 Filter sterilize
Magnesium Chloride Solution:
Compositionper 10.0mL:
MgCl2·6H2O 0.4g
Preparation of Magnesium Chloride Solution: Add 0.4g of MgCl2·6H2O to distilled/deionized water and bring volume to 10.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Cool
to room temperature Sparge with 100% N2
Calcium Chloride Solution:
Compositionper 10.0mL:
CaCl2·2H2O 0.05g
Preparation of Calcium Chloride Solution: Add CaCl2·2H2O
to distilled/deionized water and bring volume to 10.0mL Mix thor-oughly Autoclave for 15 min at 15 psi pressure–121°C Cool to room temperature Sparge with 100% N2
Seven Vitamin Solution:
Compositionper liter:
Pyridoxine·HCl 300.0mg Nicotinic acid 200.0mg Thiamine·HCl 200.0mg Calcium DL-pantothenate 100.0mg Cyanocobalamine 100.0mg
p-Aminobenzoic acid 80.0mg
D(+)-Biotin 20.0mg
Trang 5Thiomicrospira denitrificans Agar 1779
Preparation of Seven Vitamin Solution: Add components to
dis-tilled/deionized water and bring volume to 1.0L Mix thoroughly Filter
sterilize Sparge with 100% N2
Trace Elements Solution SL-10 with EDTA:
Compositionper liter:
FeCl2·4H2O 1.5g
Na2-EDTA 0.5g
CoCl2·6H2O 190.0mg
MnCl2·4H2O 100.0mg
ZnCl2 70.0mg
Na2MoO4·2H2O 36.0mg
NiCl2·6H2O 24.0mg
H3BO3 6.0mg
CuCl2·2H2O 2.0mg
HCl (25% solution) 10.0mL
Preparation of Trace Elements Solution SL-10 with EDTA:
Add FeCl2·4H2O to 10.0mL of HCl solution Mix thoroughly Add
dis-tilled/deionized water and bring volume to 1.0L Add remaining
com-ponents Mix thoroughly Sparge with 100% N2 Autoclave for 15 min
at 15 psi pressure–121°C Adjust pH to 7.0
Preparation of Medium: Add components, except magnesium
chloride, thiosulfate, bicarbonate, and vitamin solutions, to distilled/
deionized water and bring volume to 970.0mL Mix thoroughly Sparge
with 80% N2 + 20% CO2 gas mixture for at least 30 min to remove
dis-solved oxygen and to saturate the solution with CO2 Dispense into
an-aerobic culture vessels (e.g., Balch tubes) to 1/2 volume under air
atmosphere Close vials with butyl rubber septa to prevent free
ex-change of oxygen with the external atmosphere Autoclave for 15 min
at 15 psi pressure–121°C Cool to room temperature Aseptically add
magnesium chloride, thiosulfate, bicarbonate, and vitamin solutions
Mix thoroughly Adjust pH to 7.5–7.8 using a sterile solution of sodium
carbonate (5% w/v)
Use: For the cultivation of Thiohalomonas nitratireducens
Thiol HiVeg Broth
Compositionper liter:
Plant peptone No 3 10.0g
Thiol compound 8.0g
NaCl 5.0g
Yeast extract 5.0g
Glucose 1.0g
p-Aminobenzoic acid (PABA) 0.05g
pH 7.1 ± 0.2 at 25°C
Source: This medium is available as a premixed powder from
Hi-Media
Preparation of Medium: Add components to distilled/deionized
water and bring volume to 1.0L Mix thoroughly Gently heat and bring
to boiling Distribute into tubes or flasks For neutralization of
penicil-lin, distribute medium into tubes to a depth of 60.0mm For
neutraliza-tion of streptomycin, distribute medium into tubes in shallow layers
Autoclave for 15 min at 15 psi pressure–121°C
Use: For the cultivation of bacteria from body fluids and other
mate-rials containing penicillin, streptomycin, or sulfonamides Also used
for the cultivation and maintenance of Bifidobacterium species.
Thiol HiVeg Medium
Compositionper liter:
Plant peptone No 3 10.0g
Thiol compound 8.0g
Yeast extract 5.0g NaCl 5.0g Agar 1.0g Glucose 1.0g
p-Aminobenzoic acid (PABA) 0.05g
pH 7.1 ± 0.2 at 25°C
Source: This medium is available as a premixed powder from Hi-Media
Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Gently heat and bring
to boiling Distribute into tubes or flasks For neutralization of penicil-lin, distribute medium into tubes to a depth of 60 mm For neutraliza-tion of streptomycin, distribute medium into tubes in shallow layers Autoclave for 15 min at 15 psi pressure–121°C
Use: For the cultivation of bacteria from body fluids and other mate-rials containing penicillin, streptomycin, or sulfonamides Also used
for the cultivation and maintenance of Bifidobacterium species.
Thiol Medium
Composition per liter:
Proteose peptone No 3 10.0g Thiol complex 8.0g Yeast extract 5.0g NaCl 5.0g Glucose 1.0g Agar 1.0g
p-Aminobenzoic acid 0.05g
pH 7.1 ± 0.2 at 25°C
Source: This medium is available as a premixed powder from BD Di-agnostic Systems
Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Gently heat and bring
to boiling Distribute into tubes or flasks For neutralization of penicil-lin, distribute medium into tubes to a depth of 60mm For neutraliza-tion of streptomycin, distribute medium into tubes in shallow layers Autoclave for 15 min at 15 psi pressure–121°C
Use: For the cultivation of bacteria from body fluids and other mate-rials containing penicillin, streptomycin, or sulfonamides Also used
for the cultivation and maintenance of Bifidobacterium species.
Thiomicrospira denitrificans Agar
Composition per 1001.0mL:
Solution A 940.0mL Solution B 40.0mL Solution C 20.0mL Solution D 1.0mL
pH 7.0 ± 0.2 at 25°C
Solution A:
Compositionper 940.0mL:
Agar 15.0g
KH2PO4 2.0g KNO3 2.0g
NH4Cl 1.0g MgSO4·7H2O 0.8g Trace elements solution SL-4 2.0mL
Preparation of Solution A: Add components to distilled/deionized water and bring volume to 940.0mL Mix thoroughly Gently heat and
Trang 61780 Thiomicrospira denitrificans Broth
bring to boiling Adjust pH to 7.0 with NaOH Autoclave for 15 min at
15 psi pressure–121°C Cool to 45°–50°C
Trace Elements Solution SL-4:
Composition per liter:
EDTA 0.5g
FeSO4·7H2O 0.2g
Trace elements solution SL-6 100.0
Preparation of Trace Elements Solution SL-4: Add components
to distilled/deionized water and bring volume to 1.0L Mix thoroughly
Trace Elements Solution SL-6:
Compositionper liter:
H3BO3 0.3g
CoCl2·6H2O 0.2g
ZnSO4·7H2O 0.1g
MnCl2·4H2O 0.03g
Na2MoO4·H2O 0.03g
NiCl2·6H2O 0.02g
CuCl2··2H2O 0.01g
Preparation of Trace Elements Solution SL-6: Add components
to distilled/deionized water and bring volume to 1.0L Mix thoroughly
Adjust pH to 3.4
Solution B:
Composition per 40.0mL:
Na2S2O3·5H2O 5.0g
Preparation of Solution B: Add Na2S2O3·5H2O to
distilled/deion-ized water and bring volume to 40.0mL Mix thoroughly Autoclave for
15 min at 15 psi pressure–121°C Cool to 45°–50°C
Solution C:
Composition per 20.0mL:
NaHCO3 1.0g
Preparation of Solution C: Add NaHCO3 to distilled/deionized
water and bring volume to 20.0mL Mix thoroughly Filter sterilize
Solution D:
Composition per liter:
FeSO4·7H2O 2.0mg
H2SO4 (0.1N solution) 1.0mL
Preparation of Solution D: Add FeSO4·7H2O to 1.0mL of 0.1N
H2SO4 solution Mix thoroughly Autoclave for 15 min at 15 psi
pres-sure–121°C Cool to 45°–50°C
Preparation of Medium: Aseptically add 40.0mL of sterile
solu-tion B, 20.0mL of sterile solusolu-tion C, and 1.0mL of sterile solusolu-tion D to
940.0mL of sterile solution A Mix thoroughly Aseptically and
anaer-obically distribute into sterile tubes under 100% N2
Use: For the cultivation and maintenance of Thiomicrospira
denitrifi-cans.
Thiomicrospira denitrificans Broth
Composition per 1001.0mL:
Solution A 940.0mL
Solution B 40.0mL
Solution C 20.0mL
Solution D 1.0mL
pH 7.0 ± 0.2 at 25°C
Solution A:
Compositionper 940.0mL:
KH2PO4 2.0g KNO3 2.0g
NH4Cl 1.0g MgSO4·7H2O 0.8g Trace elements solution SL-4 2.0mL
Preparation of Solution A: Add components to distilled/deionized water and bring volume to 940.0mL Mix thoroughly Adjust pH to 7.0 with NaOH Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C
Solution B:
Composition per 40.0mL:
Na2S2O3·5H2O 5.0g
Preparation of Solution B: Add Na2S2O3·5H2O to distilled/deion-ized water and bring volume to 40.0mL Mix thoroughly Autoclave for
15 min at 15 psi pressure–121°C Cool to 45°–50°C
Solution C:
Composition per 20.0mL:
NaHCO3 1.0g
Preparation of Solution C: Add NaHCO3 to distilled/deionized water and bring volume to 20.0mL Mix thoroughly Filter sterilize
Solution D:
Composition per liter:
FeSO4·7H2O 2.0mg
H2SO4 (0.1N solution) 1.0mL
Preparation of Solution D: Add FeSO4·7H2O to 1.0mL of 0.1N
H2SO4 solution Mix thoroughly Autoclave for 15 min at 15 psi pres-sure–121°C Cool to 45°–50°C
Trace Elements Solution SL-4:
Composition per liter:
EDTA 0.5g FeSO4·7H2O 0.2g Trace elements solution SL-6 100.0mL
Preparation of Trace Elements Solution SL-4: Add components
to distilled/deionized water and bring volume to 1.0L Mix thoroughly
Trace Elements Solution SL-6:
Compositionper liter:
H3BO3 0.3g CoCl2·6H2O 0.2g ZnSO4·7H2O 0.1g MnCl2·4H2O 0.03g
Na2MoO4·H2O 0.03g NiCl2·6H2O 0.02g CuCl2·2H2O 0.01g
Preparation of Trace Elements Solution SL-6: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Ad-just pH to 3.4
Preparation of Medium: Aseptically add 40.0mL of sterile solution
B, 20.0mL of sterile solution C, and 1.0mL of sterile solution D to 940.0mL of sterile solution A Mix thoroughly Aseptically and anaero-bically distribute into sterile tubes under 100% N2
Use: For the cultivation and maintenance of Thiomicrospira
denitrifi-cans.
Trang 7Thiomicrospira Medium 1781
Thiomicrospira denitrificans Medium
Compositionper liter:
Part I 500.0mL
Part II 500.0mL
pH 7.0–8.0 at 25°C
Part I:
Compositionper liter:
NaCl 20.0g
KNO3 4.0g
(NH4)2SO4 2.0g
MgSO4·7H2O 1.5g
K2HPO4 0.6g
KH2PO4 0.4g
FeSO4 solution 2.0mL
Trace metals solution 2.0mL
HCl, concentrated 1.0mL
Preparation of Part I: Add components to distilled/deionized
wa-ter and bring volume to 1.0L Mix thoroughly Autoclave for 15 min at
15 psi pressure–121°C Cool to 25°C
FeSO 4 Solution:
Compositionper 100.0mL:
FeSO4·7H2O 0.5g
HCl (1N solution) 100.0mL
Preparation of FeSO 4 Solution: Combine the FeSO4·7H2O and
100.0mL of HCl solution Mix thoroughly
Trace Metals Solution:
Compositionper liter:
Disodium EDTA 50.0g
NaOH 11.0g
CaCl2·2H2O 7.34g
MnCl2·2H2O 2.5g
ZnSO4·7H2O 2.2g
CoCl2·6H2O 0.5g
(NH4)6Mo7O24·4H2O 0.5g
CuSO4·5H2O 0.2g
Preparation of Trace Metals Solution: Add EDTA to distilled/
deionized water and bring volume to 500.0mL Mix thoroughly Adjust
pH to 6.0 with NaOH Add remaining components, one by one
Main-tain the pH at 6.0 After dissolution of all the salts, adjust the pH to 4.0
with HCl Store at 4°C
Part II:
Compositionper liter:
Na2S2O3·5H2O 10.0g
NaHCO3 3.0g
NaOH 0.05g
Preparation of Part II: Add components to distilled/deionized
wa-ter and bring volume to 1.0L Mix thoroughly Autoclave for 15 min at
15 psi pressure–121°C Cool to 25°C
Preparation of Medium: Aseptically combine 500.0mL of sterile
part I and 500.0mL of sterile part II Mix thoroughly Aseptically
dis-tribute into sterile tubes or flasks
Use: For the isolation and cultivation of Thiomicrospira denitrificans.
Thiomicrospira Medium
(ATCC Medium 1036)
Compositionper liter:
NaCl 25.0g
Na2S2O3·5H2O 8.0g
MgSO4·7H2O 1.5g (NH4)2SO4 1.0g
K2HPO4 0.5g CaCl2 0.3g Vitamin B12 15.0μg Vishniac and Santer trace metals 0.2mL Bromcresol Purple (0.05% solution) 0.1mL
pH 7.2 ± 0.2 at 25°C
Vishniac and Santer Trace Metals:
Compositionper liter:
Ethylenediamine tetraacetic acid (EDTA) 50.0g ZnSO4·7H2O 22.0g CaCl2 5.54g MnCl2·4H2O 5.06g FeSO4·7H2O 4.99g CoCl2·6H2O 1.61g CuSO4·5H2O 1.57g (NH4)6Mo7O24·4H2O 1.1g
Preparation of Vishniac and Santer Trace Metals: Add com-ponents to distilled/deionized water and bring volume to 1.0L Adjust
pH to 6.0 with KOH Mix thoroughly
Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.2 Fil-ter sFil-terilize Aseptically distribute into sFil-terile tubes or flasks
Use: For the cultivation and maintenance of Thiomicrospira species.
Thiomicrospira Medium
(ATCC Medium 1422)
Compositionper liter:
NaCl 25.1g Tris·HCl 3.07g
Na2S2O3·5H2O 2.48g MgSO4·7H2O 1.5g (NH4)2SO4 1.0g
KH2PO4 0.42g CaCl2·2H2O 0.29g NaHCO3 0.2g Phenol Red (0.5% solution) 1.0mL Vishniac and Santer trace metals 0.2mL
pH 7.5 ± 0.2 at 25°C
Vishniac and Santer Trace Metals:
Compositionper liter:
Ethylenediamine tetraacetic acid (EDTA) 50.0g ZnSO4·7H2O 22.0g CaCl2 5.54g MnCl2·4H2O 5.06g FeSO4·7H2O 4.99g CoCl2·6H2O 1.61g CuSO4·5H2O 1.57g (NH4)6Mo7O24·4H2O 1.1g
Preparation of Vishniac and Santer Trace Metals: Add com-ponents to distilled/deionized water and bring volume to 1.0L Adjust
pH to 6.0 with KOH Mix thoroughly
Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.5 Fil-ter sFil-terilize Aseptically distribute into sFil-terile tubes or flasks
Use: For the cultivation and maintenance of Thiomicrospira species.
Trang 81782 Thiomicrospira pelophila Medium
Thiomicrospira pelophila Medium
Compositionper liter:
NaCl 25.0g
Agar 10.0g
Na2S2O3·5H2O 5.0–8.0g
MgSO4·7H2O 1.5g
(NH4)2SO4 1.0g
K2HPO4 0.5g
CaCl2 0.3g
Vitamin B12 0.15mg
Trace metals solution 0.2mL
Trace Metals Solution:
Compositionper liter:
Disodium EDTA 50.0g
NaOH 11.0g
CaCl2·2H2O 7.34g
FeSO4·7H2O 5.0g
MnCl2·2H2O 2.5g
ZnSO4·7H2O 2.2g
CoCl2·6H2O 0.5g
(NH4)6Mo7O24·4H2O 0.5g
CuSO4·5H2O 0.2g
Preparation of Trace Metals Solution: Add EDTA to distilled/
deionized water and bring volume to 500.0mL Mix thoroughly Adjust
pH to 6.0 with NaOH Add remaining components, one by one
Main-tain the pH at 6.0 After dissolution of all the salts, adjust the pH to 4.0
with HCl Store at 4°C
Preparation of Medium: Add components to distilled/deionized
water and bring volume to 1.0L Mix thoroughly Gently heat and bring
to boiling Distribute into tubes or flasks Autoclave for 15 min at 15
psi pressure–121°C Pour into sterile Petri dishes or leave in tubes
Use: For the cultivation of Thiomicrospira pelophila.
Thiomicrospira pelophila Medium
Compositionper 1000.2mL:
NaCl 25.0g
MgSO4·7H2O 1.5g
(NH4)2SO4 1.0g
CaCl2·2H2O 0.42g
Bromthymol Blue 4.0mg
K2HPO4 solution 100.0mL
Na2S2O3·5H2O solution 100.0mL
Vitamin B12 solution 10.0mL
Trace elements solution 0.2mL
Na2CO3 solution variable
pH 7.2 ± 0.2 at 25°C
K 2 HPO 4 Solution:
Compositionper 100.0mL:
K2HPO4 0.5g
Preparation of K 2 HPO 4 Solution: Add K2HPO4 to
distilled/de-ionized water and bring volume to 100.0mL Mix thoroughly
Auto-clave for 15 min at 15 psi pressure–121°C
Na 2 S 2 O 3 Solution:
Compositionper 10.0mL:
Na2S2O3 ·5H2O 5.0g
Preparation of Na 2 S 2 O 3 Solution: Add Na2S2O3·5H2O to
dis-tilled/deionized water and bring volume to 10.0mL Mix thoroughly
Autoclave for 15 min at 15 psi pressure–121°C
Vitamin B 12 Solution:
Compositionper 10.0mL:
Vitamin B12 15.0mg
Preparation of Vitamin B 12 Solution: Add vitamin B12 to dis-tilled/deionized water and bring volume to 10.0mL Mix thoroughly Filter sterilize
Trace Elements Solution:
Compositionper liter:
Disodium EDTA 50.0g ZnSO4·7H2O 22.0g CaCl2·2H2O 5.54g MnCl2·4H2O 5.06g FeSO4·7H2O 5.0g CoCl2·6H2O 1.61g CuSO4·5H2O 1.57g (NH4)6Mo7O24·4H2O 1.1g
Preparation of Trace Elements Solution : Add components to
distilled/deionized water and bring volume to 1.0L Mix thoroughly
Na 2 CO 3 Solution:
Compositionper 100.0mL:
Na2CO3 0.4g
Preparation of Na 2 CO 3 Solution: Add Na2CO3 to distilled/de-ionized water and bring volume to 100.0mL Mix thoroughly Auto-clave for 15 min at 15 psi pressure–121°C
Preparation of Medium: Add components, except K2HPO4 solu-tion, Na2S2O3·5H2O solution, vitamin B12 solution, and Na2CO3 solu-tion, to distilled/deionized water and bring volume to 790.0mL Mix thoroughly Adjust pH to 7.2 Autoclave for 15 min at 15 psi pressure– 121°C Aseptically add 100.0mL of sterile K2HPO4 solution, 100.0mL
of sterile Na2S2O3·5H2O solution, and 10.0mL of sterile vitamin B12 solution Mix thoroughly Aseptically adjust pH to 7.2 with the appro-priate volume of sterile Na2CO3 solution Aseptically distribute into sterile tubes or flasks
Use: For the cultivation and maintenance of Thiomicrospira
pelo-phila.
Thiomonas delicata Medium
(DSMZ Medium 1037)
Composition per liter:
Na2S2O3·5H2O 5.0g
Na2HPO4 4.5g
K2HPO4 1.5g Yeast extract 1.0g Na-aspartate 1.0g
NH4Cl 0.3g MgSO4·7H2O 0.1g
pH 6.0 ± 0.2 at 25°C
Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 6.0 Dispense into culture vessels Autoclave for 15 min at 15 psi pressure– 121°C
Use: For the cultivation of Thiomonas delicata.
Thiomonas delicata Medium
(DSMZ Medium 1037)
Composition per liter:
Na2S2O3·5H2O 5.0g
Na2HPO4 4.5g
Trang 9Thiorhodococcus Medium 1783
K2HPO4 1.5g
Yeast extract 1.0g
NH4Cl 0.3g
MgSO4·7H2O 0.1g
pH 6.0 ± 0.2 at 25°C
Preparation of Medium: Add components to distilled/deionized
water and bring volume to 1.0L Mix thoroughly Adjust pH to 6.0
Dispense into culture vessels Autoclave for 15 min at 15 psi pressure–
121°C
Use: For the cultivation of Thiomonas perometabolis.
Thiophaeococcus mangrovi Medium
(DSMZ Medium 1162)
Composition per liter:
NaCl 20.0g
Sodium pyruvate 3.0g
MgSO4·7H2O 2.0g
NH4Cl 0.64g
K2HPO4 0.5g
Yeast extract 0.4g
CaCl2·2H2O 0.15g
Bicarbonate solution 10.0mL
Sulfide solution 10.0mL
Thiosulfate solution 10.0mL
Trace element solution SL-8 1.0mL
Vitamin solution 1.0mL
pH 7.5 ± 0.2 at 25°C
Sulfide Solution :
Compositionper 10.0mL:
Na2S·9H2O 1.0g
Preparation of Sulfide Solution: Add Na2S·9H2O to
distilled/de-ionized water and bring volume to 10.0mL Mix thoroughly Sparge
with 100% N2 Filter sterilize
Thiosulfate Solution:
Composition per10.0mL:
Na2S2O3·5H2O 1.5g
Preparation of Thiosulfate Solution: Add Na2S2O3·5H2O to
dis-tilled/deionized water and bring volume to 10.0mL Mix thoroughly
Sparge with 100% N2 Filter sterilize
Bicarbonate Solution :
Compositionper 10.0mL:
NaHCO3 1.0g
Preparation of Bicarbonate Solution: Add NaHCO3 to distilled/
deionized water and bring volume to 10.0mL Mix thoroughly Sparge
with a gas mixture of 80% N2 + 20% CO2 Filter sterilize
Vitamin Solution:
Compositionper 10.0mL:
Vitamin B12 2.0mg
Preparation of Vitamin Solution: Add Vitamin B12 to distilled/
deionized water and bring volume to 10.0mL Mix thoroughly Filter
sterilize
Trace Elements Solution SL-8:
Composition per liter:
Disodium EDTA 5.2g
FeCl2·4H2O 1.5g
CoCl2·6H2O 0.19g
MnCl2·4H2O 0.1g ZnCl2 0.07g
H3BO3 0.06g NaMoO4·2H2O 0.04g CuCl2·2H2O 0.02g NiCl2·6H20 0.02g
Preparation of Trace Elements Solution SL-8: Add compo-nents to distilled/deionized water and bring volume to 1.0L Mix thor-oughly
Preparation of Medium: Add components, except sulfide, bicar-bonate, thiosulfate, and vitamin solutions, to distilled/deionized water and bring volume to 970.0mL Mix thoroughly Sparge with 80% N2 + 20% CO2 gas mixture Dispense into culture vessels Autoclave for 15 min at 15 psi pressure–121°C Cool to room temperature Aseptically add sulfide, thiosulfate, bicarbonate, and vitamin solutions Mix thor-oughly Adjust pH to 7.5
Use: For the cultivation of Thiophaeococcus mangrovi.
Thiophene-2-Carboxylic Acid Hydrazide
Thiorhodococcus Medium
(DSMZ Medium 28a)
Composition per 5.0L:
Solution A 4.0L Solution B 860.0mL Solution E 100.0mL Solution F 20.0mL Solution C 5.0mL Solution D 5.0mL
pH 7.3 at 25°C
Solution A:
Composition per 4.0L:
NaCl 100.0g MgCl2·6H2O 5.0g MgSO4 2.5g
Na2S2O3·5H2O 2.5g
KH2PO4 1.7g
NH4Cl 1.7g KCl 1.7g CaCl2·2H2O 1.25g
Preparation of Solution A: Add components to 4.0L distilled wa-ter Mix thoroughly Autoclave for 45 min at 15 psi pressure–121°C in
a 5-liter special bottle or flask with four openings at the top, together with a teflon-coated magnetic bar In this 5-liter bottle, two openings for tubes are in the central, silicon rubber stopper; one is a short, gas-inlet tube with a sterile cotton filter, and the other is an outlet tube for medium, which reaches the bottom of the vessel at one end and has, at the other end, a silicon rubber tube with a pinch cock and a bell for aseptic dispensing of the medium into bottles The other two openings have gas-tight screw caps; one of these openings is for the addition of sterile solutions and the other serves as a gas outlet After autoclaving, cool to room temperature under a 100% N2 atmosphere with a positive pressure of 0.05– 0.1 atm (a manometer for low pressure will be re-quired) Saturate the cold medium with CO2 by magnetic stirring for 30 min under a CO2 atmosphere of 0.05–0.1 atm
Solution B:
Distilled water 860.0mL
Trang 101784 Thiosphaera Agar
Preparation of Solution B: Autoclave distilled water for 15 min at
15 psi pressure–121°C in a cotton-stoppered Erlenmeyer flask Cool to
room temperature under an atmosphere of N2 in an anaerobic jar
Solution C:
Compositionper 100.0mL:
Vitamin B12 2.0mg
Preparation of Solution C: Add vitamin B12 to distilled/deionized
water and bring volume to 100.0mL Mix thoroughly Sparge under
100% N2 gas for 3 min Filter sterilize Store under N2 gas
Solution D:
Compositionper liter:
Disodium ethylendiamine tetraacetate (Disodium EDTA) 3.0g
FeSO4·7H2O 1.1g
H3BO3 0.3g
CoCl2·6H2O 0.19g
MnCl2·2H2O 50.0mg
ZnCl2 42.0mg
NiCl2·6H2O 24.0mg
Na2MoO4·2H2O 18.0mg
CuCl2·2H2O 2.0mg
Preparation of Solution D : Add components to distilled/deionized
water and bring volume to 1.0L Mix thoroughly Autoclave for 15 min
at 15 psi pressure–121°C
Solution E:
Compositionper 100.0mL:
NaHCO3 7.5g
Preparation of Solution E : Add NaHCO3 to distilled/deionized
water and bring volume to 100.0mL Mix thoroughly Sparge with
100% CO2 until saturated Filter sterilize under 100% CO2 into a
ster-ile, gas-tight 100.0mL screw-capped bottle
Solution F:
Compositionper 100.0mL:
Na2S·9H2O 10.0g
Preparation of Solution F : Add Na2S·9H2O to distilled/deionized
water in a 250.0mL screw-capped bottle fitted with a butyl rubber
sep-tum and bring volume to 100.0mL Mix thoroughly Sparge under
100% N2 gas for 3 min Autoclave for 15 min at 15 psi pressure–
121°C Cool to room temperature
Neutralized Sulfide Solution:
Compositionper 100.0mL:
Na2S·9H2O 1.5g
Preparation of Neutralized Sulfide Solution : Add Na2S·9H2O
to distilled/deionized water in a 250.0mL screw-capped bottle fitted
with a butyl rubber septum and bring volume to 100.0mL Add a
mag-netic stir bar Mix thoroughly Sparge under 100% N2 gas for 3 min
Autoclave for 15 min at 15 psi pressure–121°C Cool to room
temper-ature Adjust pH to about 7.3 with sterile 2M H2SO4 Do not open the
bottle to add H2SO4; use a sterile syringe Stir the solution continuously
to avoid precipitation of elemental sulfur The final solution should be
clear and yellow in color
Preparation of Medium: Add solutions B, C, D, E, and F to
solu-tion A through one of the screw-cap openings against a stream of either
N2 gas or, better, a mixture of 95% N2 and 5% CO2 while the medium
is magnetically stirred Adjust the pH of the medium with sterile HCl
or Na2CO3 solution (2M solutions) to pH 7.3 Distribute the medium
aseptically through the medium outlet tube into sterile, 100mL bottles
(with metal caps and autoclavable rubber seals) using the positive gas
pressure (0.05–0.1 atm) of the N2 /CO2 gas mixture Leave a small air bubble in each bottle to meet possible pressure changes The tightly sealed, screw-cap bottles can be stored for several weeks or months in the dark During the first 24 hr, the iron of the medium precipitates in the form of black flocs No other sediment should arise in the otherwise clear medium Incubate in the light at 500–1,000 lux intensity Feed pe-riodically with neutralized solution of sodium sulfide to replenish sul-fide and with other supplement solutions
Use: For the cultivation of Thiorhodococcus minor.
Thiosphaera Agar
Compositionper liter:
Agar 15.0g
Na2HPO4 4.2g
KH2PO4 1.5g
NH4Cl 0.3g MgSO4·7H2O 0.1g KNO3 0.1g Vishniac and Santer trace metals 2.0mL
pH 8.0–8.2 at 25°C
Vishniac and Santer Trace Metals:
Compositionper liter:
Ethylenediamine tetraacetic acid (EDTA) 50.0g ZnSO4·7H2O 22.0g CaCl2 5.54g MnCl2·4H2O 5.06g FeSO4·7H2O 4.99g CoCl2·6H2O 1.61g CuSO4·5H2O 1.57g (NH4)6Mo7O24·4H2O 1.1g
Preparation of Vishniac and Santer Trace Metals: Add com-ponents to distilled/deionized water and bring volume to 1.0L Adjust
pH to 6.0 with KOH Mix thoroughly
Preparation of Medium: Add components, except agar, to dis-tilled/deionized water and bring volume to 500.0mL Mix thoroughly Adjust pH to 8.0–8.2 Filter sterilize Warm to 45°–50°C Add agar to distilled/deionized water and bring volume to 500.0mL Mix
thorough-ly Gently heat and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C Aseptically combine the two ster-ile solutions Mix thoroughly Pour into sterster-ile Petri dishes or distribute into sterile tubes
Use: For the cultivation and maintenance of Thiosphaera
pantotro-pha.
Thiosphaera Broth
Compositionper liter:
Na2HPO4 4.2g
KH2PO4 1.5g
NH4Cl 0.3g MgSO4·7H2O 0.1g KNO3 0.1g Vishniac and Santer trace metals 2.0mL
pH 8.0–8.2 at 25°C
Vishniac and Santer Trace Metals:
Compositionper liter:
Ethylenediamine tetraacetic acid (EDTA) 50.0g ZnSO4·7H2O 22.0g CaCl2 5.54g MnCl2·4H2O 5.06g