Handbook of Microbiological Media, Fourth Edition part 105 pdf

M-CP Agar Base 1035
cally add 80.0mL of sterile egg yolk emulsion, 50%. Mix thoroughly.
Pour into sterile Petri dishes in 20.0mL volumes.
Use: For the cultivation of Clostridium botulinum.
McClung-Toabe Egg Yolk Agar, CDC Modified
(CDC Modified McClung-Toabe Egg Yolk Agar)
Composition per liter:
Pancreatic digest of casein 40.0g
Agar 25.0g
NaHPO
4
5.0g
Yeast extract 5.0g
D-Glucose 2.0g
NaCl 2.0g
Egg yolk emulsion 100.0mL
MgSO
4
(5% solution) 0.2mL
pH 7.4 ± 0.2 at 25°C
Egg Yolk Emulsion:
Composition
:
Chicken egg yolks 11
Whole chicken egg 1
Preparation of Egg Yolk Emulsion: Soak eggs with 1:100 dilu-
tion of saturated mercuric chloride solution for 1 min. Crack eggs. Sep-
arate yolks from whites for 11 eggs. Mix egg yolks with 1 chicken egg.
Preparation of Medium: Add components, except egg yolk emul-
sion, to distilled/deionized water and bring volume to 900.0mL. Mix
thoroughly. Gently heat while stirring and bring to boiling. Autoclave

for 15 min at 15 psi pressure–121°C. Cool to 60°C. Aseptically add
100.0mL of sterile egg yolk emulsion. Mix thoroughly. Pour into ster-
ile Petri dishes in 20.0mL volumes.
Use: For the isolation, cultivation, and differentiation of anaerobic
bacteria from foods. Bacteria that produce lecithinase appear as colo-
nies surrounded by an insoluble opaque precipitate. Bacteria that pro-
duce lipase activity appear as colonies with a sheen or “pearly” surface.
Bacteria that possess proteolytic activity appear as colonies surrounded
by a clear zone.
McClung-Toabe Egg Yolk Agar, CDC Modified
(CDC Modified McClung-Toabe Egg Yolk Agar)
Composition per liter:
Pancreatic digest of casein 40.0g
Agar 25.0g
Na
2
HPO
4
5.0g
Yeast extract 5.0g
Glucose 2.0g
NaCl 2.0g
KH
2
PO
4
1.0g
Egg yolk emulsion, 50% 100.0mL
MgSO
4

·7H
2
O (5% solution) 0.2mL
pH 7.3 ± 0.2 at 25°C
Egg Yolk Emulsion, 50%:
Composition per 100.0mL:
Chicken egg yolks 11
Whole chicken egg 1
NaCl (0.9% solution) 50.0mL
Preparation of Egg Yolk Emulsion, 50%: Soak eggs with 1:100
dilution of saturated mercuric chloride solution for 1 min. Crack eggs
and separate yolks from whites. Mix egg yolks with 1 chicken egg.
Beat to form emulsion. Measure 50.0mL of egg yolk emulsion and add
to 50.0mL of 0.9% NaCl solution. Mix thoroughly. Filter sterilize.
Warm to 45°–50°C.
Preparation of Medium: Add components—except egg yolk
emulsion, 50%—to distilled/deionized water and bring volume to
900.0mL. Mix thoroughly. Gently heat while stirring and bring to boil-
ing. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C.
Aseptically add 100.0mL of sterile egg yolk emulsion, 50%. Mix thor-
oughly. Pour into sterile Petri dishes in 15.0mL volumes.
Use: For the cultivation of a wide variety of anaerobic bacteria. For the
differentiation of anaerobic bacteria based on lecithinase production,
lipase production, and proteolytic ability. Bacteria that produce lecithi-
nase appear as colonies surrounded by a zone of insoluble precipitate.
Bacteria that produce lipase appear as colonies with a pearly iridescent
sheen. Bacteria that produce proteolytic activity appear as colonies sur-
rounded by a clear zone.
McClung Toabe HiVeg Agar Base wtih Egg Yolk
Composition per liter:

Plant peptone No. 3 40.0g
Agar 25.0g
Na
2
HPO
4
5.0g
Glucose 2.0g
NaCl 2.0g
KH
2
PO
4
1.0g
MgSO
4
0.1g
Egg yolk emulsion, 50% 100.0mL
pH 7.3 ± 0.2 at 25°C
Source: This medium, without egg yolk emulsion, is available as a
premixed powder from HiMedia.
Egg Yolk Emulsion, 50%:
Composition per 100.0mL:
Chicken egg yolks 11
Whole chicken egg 1
NaCl (0.9% solution) 50.0mL
Preparation of Egg Yolk Emulsion, 50%: Soak eggs with 1:100
dilution of saturated mercuric chloride solution for 1 min. Crack eggs
and separate yolks from whites. Mix egg yolks with 1 chicken egg.
Beat to form emulsion. Measure 50.0mL of egg yolk emulsion and add

to 50.0mL of 0.9% NaCl solution. Mix thoroughly. Filter sterilize.
Warm to 45°–50°C.
Preparation of Medium: Add components, except egg yolk emul-
sion, 50%, to distilled/deionized water and bring volume to 900.0mL.
Mix thoroughly. Gently heat while stirring and bring to boiling. Auto-
clave for 20 min at 15 psi pressure–121°C. Cool to 50°–55°C. Asepti-
cally add 100.0mL of sterile egg yolk emulsion, 50%. Mix thoroughly.
Pour into sterile Petri dishes in 15.0mL volumes.
Use: For the isolation and cultivation of Clostridium perfringens in
foods.
M-CP Agar Base
Composition per liter:
Tryptose 30.0g
Yeast extract 20.0g
Agar 15.0g
Sucrose 5.0g
L-Cysteine·HCl·H
2
O 1.0g
MgSO
4
·7H
2
O 0.1g
FeCl
3
·6H
2
O 0.09g
Indoxyl β-

D-glucoside 0.06g
© 2010 by Taylor and Francis Group, LLC
1036 M-CP HiVeg Agar Base with Phenolphthalein Diphosphate
Bromcresol Purple 0.04g
Selective supplement solution B 20.0mL
Selective supplement solution A 10.0mL
pH 7.6 ± 0.2 at 25°C
Source: This medium is available from HiMedia.
Selective Supplement Solution A:
Composition
per 10.0mL:
D-Cycloserine 400.0mg
Polymyxin B sulfate 25.0mg
Preparation of Selective Supplement Solution A: Add compo-
nents to distilled/deionized water and bring volume to 10.0mL. Mix
thoroughly. Filter sterilize.
Selective Supplement Solution B:
Composition
per 20.0mL:
Phenolphthalein diphosphate 0.1g
Preparation of Selective Supplement Solution B: Add phenol-
phthalein diphosphate to distilled/deionized water and bring volume to
20.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium: Add components, except selective sup-
plement solutions A and B, to distilled/deionized water and bring vol-
ume to 970.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi
pressure–121°C. Cool to 50°C. Aseptically add selective supplement
solutions A and B. Mix thoroughly. Pour into Petri dishes or aseptically
distribute into sterile tubes.
Use: Recommended by the Directive of the Council of the European

Union 98/83/EC for the isolation and enumeration of Clostridium per-
fringens from water samples using the membrane filtration technique.
M-CP HiVeg Agar Base
with Phenolphthalein Diphosphate
Composition per liter:
Plant hydrolysate No. 1 30.0g
Agar 15.0g
L-Cysteine·HCl 1.0g
MgSO
4
·7H
2
O 0.1g
FeCl
3
·6H
2
O 0.09
Indoxyl β-D-glucoside 0.06
Bromcresol Purple 0.04
Sucrose 5.0g
Yeast extract 20.0g
Phenolphthalein diphosphate solution 10.0mL
pH 7.2 ± 0.2 at 25°C
Source: This medium, without phenolphthalein diphosphate, is avail-
able as a premixed powder from HiMedia.
Phenolphthalein Diphosphate Solution:
Composition
per 10.0mL:
Phenolphthalein diphosphate 2.0g

Preparation of Phenolphthalein Diphosphate Solution: Add
phenolphthalein diphosphate to distilled/deionized water and bring
volume to 10.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium: Add components, except phenolphtha-
lein diphosphate solution, to distilled/deionized water and bring vol-
ume to 990.0mL. Mix thoroughly. Gently heat while stirring until
boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45-
50°C. Aseptially add 10.0mL of sterile phenolphthalein diphosphate
solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into
sterile tubes.
Use: For the cultivation and identification of Providencia stuartii.
m-CP Medium
Composition per liter:
Tryptose 30.0g
Yeast extract 20.0g
Agar 15.0g
Sucrose 5.0g
L-Cysteine·HCl·H
2
O 1.0g
MgO
4
·7H
2
O 0.1g
Bromcresol Purple 0.04g
Phenolphthalein biphosphate tetrazolium
salt solution 10.0mL
Selective supplement solution 4.0mL
Indoxyl-β-D-glucoside solution 4.0mL

Ferric chloride solution 1.0mL
pH 7.6 ± 0.2 at 25°C
Selective Supplement Solution:
Composition
per 4.0mL:
D-Cycloserine 0.4g
Polymyxin B sulfate 25.0mg
Preparation of Selective Supplement Solution: Add compo-
nents to 4.0mL of distilled/deionized water. Mix thoroughly. Filter ster-
ilize.
Phenolphthalein Biphosphate Tetrazolium
Salt Solution:
Composition
per 10.0mL:
Phenolphthalein biphosphate tetrazolium
salt 25.0mg
Preparation of Phenolphthalein Biphosphate Tetrazolium
Salt Solution:
Add phenolphthalein biphosphate tetrazolium salt to
10.0mL of distilled/deionized water. Mix thoroughly. Filter sterilize.
Indoxyl-β-D-glucoside Solution:
Composition
per 10.0mL:
Indoxyl-β-
D-glucoside 0.45g
Preparation of Indoxyl-β-D-glucoside Solution: Add indoxyl-
β-
D-glucoside to 10.0mL of distilled/deionized water. Mix thoroughly.
Filter sterilize.
Ferric Chloride Solution:

Composition
per 4.0mL:
FeCl
3
·6H
2
O 30.0mg
Preparation of Ferric Chloride Solution: Add FeCl
3
·6H
2
O to
4.0mL of distilled/deionized water. Mix thoroughly. Filter sterilize.
Preparation of Medium: Add components, except selective sup-
plement solution, phenolphthalein biphosphate tetrazolium salt solu-
tion, indoxyl-β-
D-glucoside solution, and ferric chloride solution, to
distilled/deionized water and bring volume to 981.0mL. Mix thorough-
ly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C.
Aseptically add 4.0mL of selective supplement solution. Mix thor-
oughly. Aseptically add 10.0mL phenolphthalein biphosphate tetrazo-
lium salt solution, 4.0mL indoxyl-β-
D-glucoside solution, and 1.0mL
ferric chloride solution. Mix thoroughly. Pour into sterile Petri dishes or
aseptically distribute into tubes.
Use: A selective, chromogenic medium for the rapid identification and
enumeration of Clostridium perfringens in water samples, including
water used in food and beverage production.
© 2010 by Taylor and Francis Group, LLC
MD1- Medium 1037

MCP Medium
(Modified MacConkey Medium)
(MacConkey Phosphatase Medium)
Composition per liter:
Pancreatic digest of gelatin 17.0g
Agar 13.5g
Lactose 10.0g
NaCl 5.0g
Bile salts 1.5g
Pancreatic digest of casein 1.5g
Peptic digest of animal tissue 1.5g
Na
2
HPO
4
0.6g
Glucose 0.2g
Methyl Blue 0.07g
Neutral Red 0.03g
Crystal Violet 1.0mg
Phenolphthalein diphosphate solution 10.0mL
pH 7.2 ± 0.2 at 25°C
Source: This medium, without phenolphthalein diphosphate, is avail-
able as a premixed powder from HiMedia.
Phenolphthalein Diphosphate Solution:
Composition
per 10.0mL:
Phenolphthalein diphosphate 2.0g
Preparation of Phenolphthalein Diphosphate Solution: Add
phenolphthalein diphosphate to distilled/deionized water and bring

volume to 10.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium: Add components, except phenolphtha-
lein diphosphate solution, to distilled/deionized water and bring vol-
ume to 990.0mL. Mix thoroughly. Gently heat while stirring until
boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45-
50°C. Aseptially add 10.0mL of sterile phenolphthalein diphosphate
solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into
sterile tubes.
Use: For the cultivation and identification of Providencia stuartii.
MD Medium
Composition per liter:
Agar 20.0g
L-Malic acid 20.0g
Pancreatic digest of casein 10.0g
D-Glucose 5.0g
Casamino acids 3.0g
Pancreatic digest of soybean meal 1.5g
Tween™ 80 1.0g
Yeast extract 1.0g
Bromcresol Green solution 20.0mL
pH 7.0 ± 0.2 at 25°C
Bromcresol Green Solution:
Composition
per 30.0mL:
Bromcresol Green 0.1g
NaOH (0.01N solution) 30.0mL
Preparation of Bromcresol Green Solution: Add Bromcresol
Green to 30.0mL of NaOH solution. Mix thoroughly. Filter sterilize.
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. Adjust pH to 7.0 with 10N
KOH. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile
Petri dishes or leave in tubes.
Use: For the isolation and cultivation of Salmonella species from
foods.
MD 1 Medium
Composition per liter:
Pancreatic digest of casein 3.0g
MgSO
4
·7H
2
O 2.0g
CaCl
2
0.5g
Trace elements solution 1.0mL
Vitamin B
12
solution 1.0mL
Trace Elements Solution:
Composition per liter:
EDTA 8.0g
MnCl
2
·4H
2
O 0.1g
CoCl
2

0.02g
KBr 0.02g
ZnCl
2
0.02g
CuSO
4
0.01g
H
3
BO
3
0.01g
NaMoO
4
·2H
2
O 0.01g
BaCl
2
5.0mg
LiCl 5.0mg
SnCl
2
·2H
2
O 5.0mg
Preparation of Trace Elements Solution: Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Vitamin B

12
Solution:
Composition per 10.0mL:
Vitamin B
12
5.0mg
Preparation of Vitamin B
12
Solution: Add vitamin B
12
to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
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.
Use: For the cultivation of myxobacteria.
MD1- Medium
(DSMZ Medium 1118)
Composition per liter:
Casitone 3.0g
MgSO
4
·7H
2
O 2.0g
CaCl
2
·2H
2
O 0.7g

Vitamin solution 10.0mL
Trace elements solution SL-4 1.0mL
pH 7.1 ± 0.2 at 25°C
Trace Elements Solution SL-4:
Composition
per liter:
EDTA 0.5g
FeSO
4
·7H
2
O 0.2g
Trace elements solution SL-6 100.0mL
Trace Elements Solution SL-6:
Composition
per liter:
MnCl
2
·4H
2
O 0.5g
H
3
BO
3
0.3g
CoCl
2
·6H
2

O 0.2g
ZnSO
4
·7H
2
O 0.1g
Na
2
MoO
4
·2H
2
O 0.03g
© 2010 by Taylor and Francis Group, LLC
1038 MDPA with Calcium Carbonate
NiCl
2
·6H
2
O 0.02g
CuCl
2
·2H
2
O 0.01g
Preparation of Trace Elements Solution SL-6: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Trace Elements Solution SL-4: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

Filter sterilize.
Vitamin Solution:
Composition
per 10.0ml:
Vitamin B
12
0.5mg
Preparation of Vitamin Solution: Add components to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Filter
sterilize.
Preparation of Medium: Add components, except vitamin solu-
tion, to distilled/deionized water and bring volume to 990.0mL. Mix
thoroughly. Adjust pH to 7.1. Distribute into tubes or flasks. Gently
heat while stirring and bring to boiling. Mix thoroughly. Autoclave for
15 min at 15 psi pressure–121°C. Aseptically add 10.0mL sterile vita-
min solution. Mix thorougly. Aseptically distribute into sterile tubes or
flasks.
Use: For the cultivation of Myxococcus xanthus.
MDPA
See: Malt Dextrose Peptone Agar
MDPA with Calcium Carbonate
Composition per liter:
Agar 25.0g
Malt extract 20.0g
Glucose 20.0g
CaCO
3
5.0g
Peptone 1.0g
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 Dekkera species.
MDPYA4
See: Malt 4% Dextrose Peptone Yeast Agar
MDYA4
See: Malt 4% Dextrose Yeast Agar
m-E Agar
See: E Agar
Me15% MH Agar
(DSMZ Medium 582)
Composition per liter:
NaCl 121.5g
Agar 20.0g
MgSO
4
14.4g
MgCl
2
10.5g
Yeast extract 10.0g
Proteose peptone no. 3 5.0g
KCl 3.0g
Glucose 1.0g
CaCl
2
0.54g
NaBr 0.039g
NaHCO

3
solution 10.0mL
pH 7.5 ± 0.2 at 25°C
NaHCO
3
Solution:
Composition
per 100.0mL:
NaHCO
3
0.9g
Preparation of NaHCO
3
Solution: Add NaHCO
3
to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 80% N
2
+ 20% CO
2
. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 50°C.
Preparation of Medium: Add components, except NaHCO
3
solu-
tion, to distilled/deionized water and bring volume to 990.0mL. Mix
thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to
50°C. Aseptically add 10.0mL NaHCO
3

solution. Mix thoroughly.
Pour into Petri dishes or aseptically distribute into sterile tubes.
Use: For the cultivation of Bacillus halophilus.
Me15% MH Medium
(DSMZ Medium 582)
Composition per liter:
NaCl 121.5g
MgSO
4
14.4g
MgCl
2
10.5g
Yeast extract 10.0g
Proteose peptone no. 3 5.0g
KCl 3.0g
Glucose 1.0g
CaCl
2
0.54g
NaBr 0.039g
NaHCO
3
solution 10.0mL
pH 7.5 ± 0.2 at 25°C
NaHCO
3
Solution:
Composition
per 100.0mL:

NaHCO
3
0.9g
Preparation of NaHCO
3
Solution: Add NaHCO
3
to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 80% N
2
+ 20% CO
2
. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 25°C.
Preparation of Medium: Add components, except NaHCO
3
solu-
tion, to distilled/deionized water and bring volume to 990.0mL. Mix
thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to
25°C. Aseptically add 10.0mL NaHCO
3
solution. Mix thoroughly.
Aseptically distribute into sterile tubes or flasks.
Use: For the cultivation of Bacillus halophilus.
MEA
See: Malt Extract Agar
Meat Extract with Peptone
(Pepted Meat Broth)
Composition per liter:

NaCl 15.0g
Peptic digest of animal tissue 10.0g
Meat extract 3.0g
pH 7.2 ± 0.2 at 25°C
© 2010 by Taylor and Francis Group, LLC
Medium A for Producing Lysates 1039
Source: This medium is available from HiMedia.
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 and maintenance of Alcaligenes species.
Meat Extract with Peptone and 1.5% Salt
Composition per liter:
NaCl 15.0g
Peptone 10.0g
Meat extract 3.0g
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.
Use: For the cultivation and maintenance of Alcaligenes species.
Meat Infusion Agar, HiVeg
(Standard Infusion Agar, HiVeg)
Composition per liter:
Agar 25.0g
Plant infusion 10.0g
Plant peptone 10.0g
NaCl 5.0g
pH 7.5 ± 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. Distribute into tubes
or flasks. Autoclave for 15 min at 15 psi pressure–121°C.
Use: For the mass cultivation of organisms for vaccine or toxin pro-
duction.
M-EC Test Agar
Composition per liter:
Agar 15.0g
Lactose 10.0g
NaCl 7.5g
Proteose peptone 5.0g
Dipotassium phosphate 3.3g
Yeast extract 3.0g
KH
2
PO
4
1.0g
Sodium lauryl sulphate 0.2g
Sodium deoxycholate 0.1g
Bromcresol Purple 0.08g
Bromphenol Red 0.08g
pH 7.3 ± 0.2 at 25°C
Source: This medium is available from HiMedia.
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. Do not autoclave. Pour into
sterile Petri dishes or leave in tubes.
Use: For the detection of Escherichia coli in water samples using the

membrane filter technique.
MED IIa
Composition per liter:
Tris buffer stock solution 10.0mL
CaCl
2
(5.0% solution) 10.0mL
MgSO
4
·7H
2
O (3.33% solution) 1.0mL
pH 7.2 ± 0.2 at 25°C
Tris Buffer Stock Solution:
Composition
per 500.0mL:
Tris(hydroxymethyl)aminomethane·HCl 35.01g
Tris(hydroxymethyl)aminomethane 3.35g
Preparation of Tris Buffer Stock Solution: Add components to
distilled/deionized water and bring volume to 500.0mL. Mix thorough-
ly. Adjust pH to 7.2.
Preparation of Medium: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes
or flasks. Autoclave for 20 min at 15 psi pressure–121°C.
Use: For the cultivation and maintenance of Vampirovibrio chlorellavo-
rus.
Medium A
Composition per liter:
D-Glucose 20.0g
Agar 20.0g

Yeast extract 10.0g
Biotin 1.0mg
Calcium pantothenate 1.0mg
pH 7.3 ± 0.2 at 25°C
Preparation of Medium: Add components, except biotin and cal-
cium pantothenate, to distilled/deionized water and bring volume to
990.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave
for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Add biotin and
calcium pantothenate to distilled/deionized water and bring volume to
10.0mL. Mix thoroughly. Filter sterilize. Aseptically add the sterile bi-
otin and calcium pantothenate solution to the cooled sterile basal me-
dium. Mix thoroughly. Pour into sterile Petri dishes or distribute into
sterile tubes.
Use: For the cultivation and maintenance of Zymomonas mobilis.
Medium A for Producing Lysates
Composition per liter:
Nutrient broth 8.0g
KCl 1.0g
MgSO
4
·7H
2
O 0.25g
MnCl
2
1.25mg
FeSO
4
(1.0mM solution) 1.0mL
Ca(NO

3
)
2
(1.0M solution) 1.0mL
pH 7.0–7.2 at 25°C
Preparation of Medium: Add components, except FeSO
4
and
Ca(NO
3
)
2
, to distilled/deionized water and bring volume to 998.0mL.
Mix thoroughly. Adjust pH to 7.0–7.2. Autoclave for 30 min at 15 psi
pressure–115°C. Cool to 45°–50°C. Prepare 1.0mM FeSO
4
solution
and 1.0M Ca(NO
3
)
2
solution separately. Filter sterilize both solutions.
Aseptically add the sterile FeSO
4
solution and sterile Ca(NO
3
)
2
solu-
tion to the cooled sterile basal medium. Mix thoroughly. Distribute into

sterile tubes or flasks.
Use: For the cultivation of microorganisms to be lysed.
© 2010 by Taylor and Francis Group, LLC
1040 Medium 2A
Medium 2A
Composition per liter:
Arginine 10.0g
NaCl 5.0g
Agar 4.0g
Peptone 1.0g
K
2
HPO
4
·3H
2
O 0.3g
Phenol Red 0.01g
pH 7.2–7.4 at 25°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. Autoclave for 15 min at 15 psi pres-
sure–121°C.
Use: For the cultivation and differentiation of Pseudomonas species
based on their production of arginine dihydrolase activity.
Medium AS4
Composition per liter:
Sucrose 80.0g
PPLO broth without Crystal Violet 500.0mL
Horse serum 200.0mL

Phenol Red (0.5% solution) 5.0mL
pH 7.2 ± 0.2 at 25°C
PPLO Broth without Crystal Violet:
Composition
per 500.0mL:
Beef heart, solids from infusion 11.53g
Peptone 2.33g
NaCl 1.15g
Source: PPLO broth without Crystal Violet is available as a premixed
powder from BD Diagnostic Systems.
Preparation of PPLO Broth without Crystal Violet: Add
components to distilled/deionized water and bring volume to 500.0mL.
Mix thoroughly. Beef heart for infusion may be substituted; 100.0g of
beef heart for infusion is equivalent to 500.0g of fresh heart tissue.
Preparation of Medium: Add components, except horse serum, to
distilled/deionized water and bring volume to 800.0mL. Mix thorough-
ly. Adjust pH to 7.2. Autoclave for 10 min at 15 psi pressure–121°C.
Cool to 45°–50°C. Aseptically add 200.0mL of noninactivated, sterile
horse serum. Mix thoroughly. Aseptically distribute into sterile tubes
or flasks.
Use: For the cultivation and maintenance of Spiroplasma melliferum.
Medium for Acetivibrio cellulolyticus
See: BC Medium
Medium for Aciduric, Thermophilic Bacillus Strains
Composition per liter:
Solution A 500.0mL
Solution B 500.0mL
pH 4.3 ± 0.2 at 25°C
Solution A:
Composition

per 500.0mL:
KH
2
PO
4
3.0g
Glucose 1.0g
Starch 1.0g
Tryptone 1.0g
Yeast extract 1.0g
MgSO
4
·7H
2
O 0.5g
CaCl
2
·2H
2
O 0.25g
(NH
4
)
2
SO
4
0.2g
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 500.0mL. Mix thoroughly. Adjust pH to 4.3.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C.

Solution B:
Composition
per 500.0mL:
Agar 20.0g
Preparation of Solution B: Add agar to distilled/deionized water
and bring volume to 500.0mL. Mix thoroughly. Autoclave for 15 min
at 15 psi pressure–121°C. Cool to 50°–55°C.
Preparation of Medium: Aseptically combine 500.0mL of solu-
tion A and 500.0mL of solution B. Mix thoroughly. Pour into sterile
Petri dishes or distribute into sterile tubes.
Use: For the cultivation of aciduric, thermophilic Bacillus strains.
Medium 2508-85-1 with Amino Acids
Composition per liter:
Agar 20.0g
Nutrient broth 8.0g
D-Glucose 5.0g
Polypeptone™ 5.0g
Yeast extract 5.0g
L-Lysine 0.1g
L-Methionine 0.05g
Diaminopimelic acid 0.05g
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 30 min at 15
psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.
Use: For the cultivation and maintenance of Escherichia coli.
Medium for Ammonia Oxidizers
Composition per liter:
MgSO
4

·7H
2
O 0.2g
(NH
4
)
2
SO
4
0.13g
K
2
HPO
4
0.09g
CaCl
2
·2H
2
O 0.02g
Chelated iron 1.0mg
MnCl
2
·4H
2
O 0.2mg
Na
2
MoO
4

·2H
2
O 0.1mg
ZnSO
4
·7H
2
O 0.1mg
CuSO
4
·5H
2
O 0.02mg
CoCl
2
·6H
2
O 2.0μg
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.
Use: For the isolation, cultivation, and enrichment of ammonia-oxi-
dizing bacteria from soil.
Medium for Ammonia Oxidizers
Composition per liter:
(NH
4
)
2
SO

4
2.0g
MgSO
4
·7H
2
O 0.2g
CaCl
2
·2H
2
O 0.02g
K
2
HPO
4
0.02g
Chelated iron 1.0mg
© 2010 by Taylor and Francis Group, LLC
Medium for Ammonia-Oxidizing Bacteria 1041
MnCl
2
·4H
2
O 0.2mg
Na
2
MoO
4
·2H

2
O 0.1mg
ZnSO
4
·7H
2
O 0.1mg
CuSO
4
·5H
2
O 0.02mg
CoCl
2
·6H
2
O 2.0μg
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.
Use: For the isolation, cultivation, and enrichment of ammonia-oxi-
dizing bacteria from soil.
Medium for Ammonia Oxidizers
Composition per liter:
K
2
HPO
4
0.5g
(NH

4
)
2
SO
4
0.5g
Phenol Red 0.5g
MgSO
4
·7H
2
O 0.05g
CaCl
2
·2H
2
O 0.02g
NaCl 0.02g
Na
2
MoO
4
·2H
2
O 2.4μg
Metals “44” 1.0mL
Metals “44”:
Composition
per 100.0mL:
ZnSO

4
·7H
2
O 1.1g
FeSO
4
·7H
2
O 0.5g
EDTA 0.25g
MnSO
4
·7H
2
O 0.154g
CuSO
4
·5H
2
O 0.04g
Co(NO
3
)
2
·6H
2
O 0.025g
Na
2
B

4
O
7
·10H
2
O 0.018g
Preparation of Metals “44”: Add a few drops of H
2
SO
4
to dis-
tilled/deionized water to inhibit precipitate formation. Add compo-
nents to acidified distilled/deionized water and bring volume to
100.0mL. Mix thoroughly.
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.
Use: For the isolation, cultivation, and enrichment of ammonia-oxi-
dizing bacteria from soil.
Medium for Ammonia Oxidizers
Composition per liter:
(NH
4
)
2
SO
4
0.5g
KH
2

PO
4
0.2g
CaCl
2
·2H
2
O 0.04g
MgSO
4
·7H
2
O 0.04g
Ferric citrate 0.5mg
Phenol Red 0.5mg
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.
Use: For the isolation, cultivation, and enrichment of ammonia-oxi-
dizing bacteria from soil.
Medium for Ammonia Oxidizers, Brackish
Composition per liter:
CaCO
3
5.0g
NH
4
Cl 0.5g
K
2

HPO
4
0.05g
Seawater 400.0mL
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.
Use: For the isolation, cultivation, and enrichment of ammonia-oxidiz-
ing bacteria from brackish specimens.
Medium for Ammonia Oxidizers, Marine
Composition per liter:
(NH
4
)
2
SO
4
1.32g
MgSO
4
·7H
2
O 0.2g
Chelated iron 0.13g
K
2
HPO
4
0.11g
CaCl

2
·2H
2
O 0.02g
ZnSO
4
·7H
2
O 0.1mg
CuSO
4
·5H
2
O 0.02mg
CoCl
2
·6H
2
O 2.0μg
MnCl
2
·4H
2
O 2.0μg
Na
2
MoO
4
·2H
2

O 1.0μg
Seawater 1.0L
Preparation of Medium: Combine components. Mix thoroughly.
Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–
121°C.
Use: For the isolation, cultivation, and enrichment of marine ammo-
nia-oxidizing bacteria.
Medium for Ammonia-Oxidizing Bacteria
Composition per liter:
(NH
4
)
2
SO
4
235.0mg
KH
2
PO
4
200.0mg
CaCl
2
·2H
2
O 40.0mg
MgSO
4
·7H
2

O 40.0mg
Iron-EDTA-Phenol Red solution 1.0mL
Na
2
CO
3
solution variable
Na
2
CO
3
Solution:
Composition
per 100.0mL:
Na
2
CO
3
5.0g
Preparation of Na
2
CO
3
Solution: Add Na
2
CO
3
to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Auto-
clave for 15 min at 15 psi pressure–121°C.

Iron-EDTA-Phenol Red Solution:
Composition
per 100.0mL:
FeSO
4
·7H
2
O 50.0mg
Sodium EDTA 50.0mg
Phenol Red 50.0mg
Preparation of Iron-EDTA-Phenol Red Solution: Add compo-
nents to distilled/deionized water and bring volume to 100.0mL. Mix
thoroughly.
Preparation of Medium: Add components, except Na
2
CO
3
solu-
tion, to distilled/deionized water and bring volume to 1.0L. Mix thor-
oughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi
pressure–121°C. Add enough sterile Na
2
CO
3
solution to turn the medi-
um pale pink. During incubation and growth of bacteria, add additional
sterile Na
2
CO
3

solution to restore the pale pink color. Growth is com-
plete when no further color change is observed.
© 2010 by Taylor and Francis Group, LLC
1042 Medium B for Sulfate Reducers
Use: For the cultivation of Nitrosolobus multiformis and Nitrosomo-
nas europaea.
Medium B for Sulfate Reducers
(Postgate’s Medium B for Sulfate Reducers)
Composition per liter:
Sodium lactate 3.5g
MgSO
4
·7H
2
O 2.0g
NH
4
Cl 1.0g
CaSO
4
1.0g
Yeast extract 1.0g
KH
2
PO
4
0.5g
FeSO
4
·7H

2
O 0.5g
Ascorbic acid 0.1g
Thioglycollic acid 0.1g
pH 7.0–7.5 at 25°C
Preparation of Medium: Add components, except ascorbic acid
and thioglycollic acid, to tap water and bring volume to 1.0L. For ma-
rine bacteria, NaCl may be added or seawater used in place of tap wa-
ter. Mix thoroughly. Adjust pH to 7.0–7.5. Thioglycolate and ascorbate
should be added immediately prior to sterilization. Distribute into
tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.
Use: For the isolation, cultivation, and maintenance of Desulfovibrio
species and Desulfotomaculum species. This medium turns black as a
result of H
2
S production due to bacterial growth.
Medium for Bacillus schlegelii
Composition per liter:
Agar 15.0g
Na
2
HPO
4
·2H
2
O 2.9g
KH
2
PO
4

2.3g
NH
4
Cl 1.0g
MgSO
4
·7H
2
O 0.5g
NaHCO
3
0.5g
CaCl
2
·2H
2
O 0.01g
MnSO
4
·H
2
O 10.0mg
Ferric ammonium citrate solution 20.0mL
Trace elements solution SL-6 5.0mL
pH 6.8 ± 0.2 at 25°C
Ferric Ammonium Citrate Solution:
Composition
per 20.0mL:
Ferric ammonium citrate 0.05g
Preparation of Ferric Ammonium Citrate Solution: Add fer-

ric ammonium citrate to distilled/deionized water and bring volume to
20.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–
121°C.
Trace Elements Solution SL-6:
Composition
per liter:
MnCl
2
·4H
2
O 0.5g
H
3
BO
3
0.3g
CoCl
2
·6H
2
O 0.2g
ZnSO
4
·7H
2
O 0.1g
Na
2
MoO
4

·2H
2
O 0.03g
NiCl
2
·6H
2
O 0.02g
CuCl
2
·2H
2
O 0.01g
Preparation of Trace Elements Solution SL-6: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Preparation of Medium: Add components, except ferric ammoni-
um citrate solution, to distilled/deionized water and bring volume to
980.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave
for 15 min at 15 psi pressure–121°C. Aseptically add 20.0mL of sterile
ferric ammonium citrate solution. Mix thoroughly. Pour into sterile Pe-
tri dishes or distribute into sterile tubes.
Use: For the chemolithotrophic growth of Bacillus schlegelii.
Medium for Bacillus stearothermophilus
Composition per 1001.0mL:
NH
4
Cl 1.0g
K
2
HPO

4
0.5g
Yeast extract 0.2g
Casamino acids 0.1g
MgSO
4
·7H
2
O 0.02g
Phenol solution 100.0mL
Trace elements solution SL-4 1.0mL
pH 7.4 ± 0.2 at 25°C
Phenol Solution:
Composition
per 100.0mL:
Phenol 0.47g
Preparation of Phenol Solution: Add phenol to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Filter ster-
ilize.
Trace Elements Solution SL-4:
Composition
per liter:
EDTA 0.5g
FeSO
4
·7H
2
O 0.2g
Trace elements solution SL-6 100.0mL
Trace Elements Solution SL-6:

Composition
per liter:
MnCl
2
·4H
2
O 0.5g
H
3
BO
3
0.3g
CoCl
2
·6H
2
O 0.2g
ZnSO
4
·7H
2
O 0.1g
Na
2
MoO
4
·2H
2
O 0.03g
NiCl

2
·6H
2
O 0.02g
CuCl
2
·2H
2
O 0.01g
Preparation of Trace Elements Solution SL-6: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Trace Elements Solution SL-4: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Filter sterilize.
Preparation of Medium: Add components, except phenol solution
and trace elements solution SL-4, to distilled/deionized water and
bring volume to 900.0mL. Mix thoroughly. Autoclave for 15 min at 15
psi pressure–121°C. Aseptically add 100.0mL of sterile phenol solu-
tion and 1.0mL of sterile trace elements solution SL-4. Mix thoroughly.
Aseptically distribute into sterile tubes or flasks.
Use: For the cultivation of Bacillus stearothermophilus.
Medium BG11 for Cyanobacteria
See: BG11 Agar and BG11 Medium
Medium BG11 for Marine Cyanobacteria
See: BG11 Marine Agar and BG11 Marine Broth
© 2010 by Taylor and Francis Group, LLC
Medium for Carbon Monoxide Oxidizers 1043
Medium with Biphenyl
(DSMZ Medium 457d)

Composition per liter:
Na
2
HPO
4
2.44g
KH
2
PO
4
1.52g
(NH
4
)
2
SO
4
0.5g
MgSO
4
·7H
2
O 0.2g
CaCl
2
·2H
2
O 0.05g
Trace elements solution SL-4 10.0mL
Biphenyl solution 25.0mL

pH 6.9 ± 0.2 at 25°C
Trace Elements Solution SL-4:
Composition per liter:
EDTA 0.5g
FeSO
4
·7H
2
O 0.2g
Trace elements solution SL-6 100.0mL
Trace Elements Solution SL-6:
Composition
per liter:
H
3
BO
3
0.3g
CoCl
2
·6H
2
O 0.2g
ZnSO
4
·7H
2
O 0.1g
MnCl
2

·4H
2
O 0.03g
Na
2
MoO
4
·H
2
O 0.03g
NiCl
2
·6H
2
O 0.02g
CuCl
2·
·2H
2
O 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.
Preparation of Trace Elements Solution SL-4: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Biphenyl Solution:
Composition per liter:
Biphenyl 10.0g
Preparation of Biphenyl Solution: Add biphenyl to 1.0L ethanol.
Mix thoroughly. Filter sterilize using a cellulose filter membrane.

Preparation of Medium: Add components, except biphenyl solu-
tion, to 1.0L distilled/deionized water. Adjust pH to 6.9. Autoclave for
15 min at 15 psi pressure–121°C. Cool to room temperature. Add an al-
iquot of the biphenyl solution to a sterile flask so that the final concen-
tration will be 0.25g/L biphenyl, and let the ethanol evaporate.
Aseptically add sterile medium to the crystal-layered flask.
Use: For the cultivation of biphenyl- utilizing bacteria.
Medium C for Sulfate Reducers
(Postgate’s Medium C for Sulfate Reducers)
Composition per liter:
Sodium lactate 6.0g
Na
2
SO
4
4.5g
NH
4
Cl 1.0g
Yeast extract 1.0g
KH
2
PO
4
0.5g
Sodium citrate·2H
2
O 0.3g
CaCl
2

·6H
2
O 0.06g
MgSO
4
·7H
2
O 0.06g
FeSO
4
·7H
2
O 0.004g
pH 7.5 ± 0.2 at 25°C
Preparation of Medium: Add components to distilled/deionized
water and bring volume to 1.0L. For marine bacteria, NaCl may be
added or sea water used in place of distilled/deionized water. Mix thor-
oughly. Adjust pH to 7.5. Distribute into tubes or flasks. Autoclave for
15 min at 15 psi pressure–121°C.
Use: For detection, culturing, and storage of Desulfovibrio species and
many Desulfotomaculum species. This medium should be used when a
clear culture medium is desired such as for chemostat culture. This
medium may be cloudy after sterilization but usually clears on cooling.
It turns black as a result of H
2
S production due to bacterial growth.
Medium for Campylobacter DSM 806
(DSMZ Medium 121)
Composition per liter:
Na-aspartate 10.0g

MgSO
4
·7H
2
O 1.0g
Yeast extract 0.2g
CaCl
2
·2H
2
O 28.0mg
Resazurin 1.0mg
Cysteine phosphate solution 100.0mL
Trace elements solution SL-10 1.0mL
pH 7.0 ± 0.2 at 25°C
Trace Elements Solution SL-10:
Composition
per liter:
FeCl
2
·4H
2
O 1.5g
CoCl
2
·6H
2
O 190.0mg
MnCl
2

·4H
2
O 100.0mg
ZnCl
2
70.0mg
Na
2
MoO
4
·2H
2
O 36.0mg
NiCl
2
·6H
2
O 24.0mg
H
3
BO
3
6.0mg
CuCl
2
·2H
2
O 2.0mg
HCl (25% solution) 10.0mL
Preparation of Trace Elements Solution SL-10: Add FeCl

2
·4H
2
O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water and bring volume to 1.0L. Add remaining components. Mix thor-
oughly. Gas under 80% N
2
+ 20% CO
2
.
Cysteine Phosphate Solution:
Composition
per 100.0mL:
K
2
HPO
4
0.75g
NaH
2
PO
4
0.25g
Cysteine-HCl·H
2
O 0.25g
Preparation of Cysteine Phosphate Solution: Add components
to 100.0mL distilled/deionized water. Mix thoroughly. Gas under 80%
N

2
+ 20% CO
2
. Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Add components, except cysteine phos-
phate solution, to distilled/deionized water and bring volume to
900.0mL. Mix thoroughly. Adjust pH to 7.0. Gently heat and bring to
boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Aseptically add 100.0mL of sterile cysteine phosphate solution. Mix
thoroughly. Aseptically distribute into sterile tubes or flasks.
Use: For the cultivation of Sulfurospirillum sp.
Medium for Carbon Monoxide Oxidizers
Composition per liter:
Agar 12.0g
Na
2
HPO
4
·12H
2
O 4.5g
NH
4
Cl 1.5g
KH
2
PO
4
0.75g
MgSO

4
·7H
2
O 0.2g
© 2010 by Taylor and Francis Group, LLC
1044 Medium for Carbon Monoxide Oxidizers
CaCl
2
·2H
2
O 0.03g
Ferric ammonium citrate 0.018g
Trace elements solution SL-6 1.0mL
pH 7.0 ± 0.2 at 25°C
Trace Elements Solution SL-6:
Composition
per liter:
MnCl
2
·4H
2
O 0.5g
H
3
BO
3
0.3g
CoCl
2
·6H

2
O 0.2g
ZnSO
4
·7H
2
O 0.1g
Na
2
MoO
4
·2H
2
O 0.03g
NiCl
2
·6H
2
O 0.02g
CuCl
2
·2H
2
O 0.01g
Preparation of Trace Elements Solution SL-6: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring
to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Pour into

sterile Petri dishes or distribute into sterile tubes. After inoculation, in-
cubate in an atmosphere of 80% CO +10% O
2
+ 10% N
2
.
Use: For the chemoautotrophic cultivation and maintenance of Alcal-
igenes species, Pseudomonas carboxydohydrogena, and other Pseudo-
monas species.
Medium for Carbon Monoxide Oxidizers
Composition per liter:
Agar 12.0g
Na
2
HPO
4
·12H
2
O 4.5g
Sodium acetate 3.0g
NH
4
Cl 1.5g
KH
2
PO
4
0.75g
MgSO
4

·7H
2
O 0.2g
CaCl
2
·2H
2
O 0.03g
Ferric ammonium citrate 0.018g
Trace elements solution SL-6 1.0mL
pH 7.0 ± 0.2 at 25°C
Trace Elements Solution SL-6:
Composition
per liter:
MnCl
2
·4H
2
O 0.5g
H
3
BO
3
0.3g
CoCl
2
·6H
2
O 0.2g
ZnSO

4
·7H
2
O 0.1g
Na
2
MoO
4
·2H
2
O 0.03g
NiCl
2
·6H
2
O 0.02g
CuCl
2
·2H
2
O 0.01g
Preparation of Trace Elements Solution SL-6: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring
to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Pour into
sterile Petri dishes or distribute into sterile tubes. After inoculation in-
cubate in air.
Use: For the chemoorganotrophic cultivation and maintenance of

Alcaligenes species, Pseudomonas carboxydohydrogena, and other
Pseudomonas species.
Medium for Chlorate Respirers
(DSMZ Medium 908)
Composition per liter:
Solution A 1.0L
Solution B 10.0mL
Solution C 10.0mL
Vitamin solution 5.0mL
Trace elements solution SL-10 1.0mL
pH 7.2 ± 0.2 at 25°C
Solution A:
Composition
per liter:
NaHCO
3
2.5g
Na-acetate 1.36g
NaClO
3
1.0g
NaH
2
PO
4
0.6g
NH
4
Cl 0.25g
KCl 0.1g

Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Sparge with 80% N
2
+ 20% CO
2
. Autoclave for 15 min at 15 psi pressure–121°C. Cool to
room temperature.
Solution B:
Composition
per 10.0mL:
MgSO
4
30.0mg
CaCl
2
·2H
2
O 10.0mg
Preparation of Solution B: Add components to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Sparge with 80%
N
2
+ 20% CO
2
. Autoclave for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
Solution C:
Na
2
MoO

4
25.0mg
Na
2
WO
4
·2H
2
O 25.0mg
Preparation of Solution C: Add components to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Sparge with 80%
N
2
+ 20% CO
2
. Autoclave for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
Vitamin Solution:
Composition
per liter:
Vitamin B
12
50.0mg
Pantothenic acid 50.0mg
Riboflavin 50.0mg
Alpha-lipoic acid 50.0mg
p-Aminobenzoic acid 50.0mg
Thiamine-HCl·2H
2
O 50.0mg

Nicotinic acid 25.0mg
Nicotinamide 25.0mg
Biotin 20.0mg
Folic acid 20.0mg
Pyridoxamine-HCl 10.0mg
Preparation of Vitamin Solution: Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Filter ster-
ilize.
Trace Elements Solution SL-10:
Composition
per liter:
FeCl
2
·4H
2
O 1.5g
H
3
BO
3
300.0mg
CoCl
2
·6H
2
O 190.0mg
MnCl
2
·4H
2

O 100.0mg
© 2010 by Taylor and Francis Group, LLC