Handbook of Microbiological Media, Fourth Edition part 47 ppsx

Costein’s LDS Test Medium 455
Bovine serum 100.0mL
Nystatin solution 1.15mL
L-Cystine (1% solution) 1.0mL
Egg yolk emulsion 10 eggs
pH 7.4 ± 0.2 at 25°C
Solution A:
Composition
per liter:
Meat extract 9.0g
Proteose peptone No. 3 9.0g
NaCl 2.7g
Glucose 1.8g
Na
2
HPO
4
·12H
2
O 1.8g
K
2
TeO
3
(2% solution) 75.0mL
L-Cystine (1% solution) 10.0mL
Caution: Potassium tellurite is toxic.
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 985.0mL. Mix thoroughly. Filter sterilize.
Egg Yolk Emulsion:
Composition

:
Chicken egg yolks 9
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 and
separate yolks from whites. Mix egg yolks with 1 chicken egg. Filter
sterilize.
Nystatin Solution:
Composition
per 10.0mL:
Nystatin 10,000U
Preparation of Nystatin Solution: Add nystatin to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Filter ster-
ilize.
L-Cystine Solution:
Composition
per 10.0mL:
L-Cystine 0.1g
Preparation of L-Cystine Solution: Add L-cystine to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Filter ster-
ilize.
Preparation of Medium: To 985.0mL of sterile solution A, asepti-
cally add the remaining components. Mix thoroughly. Aseptically dis-
tribute into sterile tubes in 2.0–3.0mL volumes.
Use: For the isolation and cultivation of Corynebacterium diphthe-
riae.
Corynebacterium Medium with Blood
(DSMZ Medium 240)
Composition per liter:
Agar 15.0g

Casein peptone, tryptic digest 10.0g
Yeast extract 5.0g
Glucose 5.0g
NaCl 5.0g
Distilled water 1000.0mL
Sheep or horse blood, defibrinated 50.0mL
pH 7.3 ± 0.2 at 25°C
Preparation of Medium: Add components, except sheep or horse
blood, to distilled/deionized water and bring volume to 950.0mL. Mix
thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at
15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add sterile
sheep or horse blood. Mix thoroughly. Pour into sterile Petri dishes or
distribute into sterile tubes.
Use: For the cultivation of Streptococcus alactolyticus, Corynebacte-
rium spp., Desemzia incerta=Brevibacterium incertum, and Moraxella
bovis.
Corynebacterium Medium CII
Composition per liter:
CaCO
3
20.0g
Agar 15.0g
Sucrose 10.0g
Yeast extract 4.0g
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 Clavibacter michiganensis.
Corynebacterium Medium with Salt

(DSMZ Medium 229)
Composition per liter:
NaCl 65.0g
Agar 15.0g
Casein peptone, tryptic digest 10.0g
Yeast extract 5.0g
Glucose 5.0g
pH 7.3 ± 0.2 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 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 Nesterenkonia halo-
bia=Micrococcus halobius.
Costein’s LDS Test Medium
Composition per liter:
Meat peptone 4.5g
Papaic digest of soybean meal 2.0g
Yeast extract 3.0g
NaCl 5.0g
D-Glucose 1.0g
L-Lysine monohydrochloride 10.0g
Na
2
S
2
O
3
0.2g
Fe(NH

4
)
2
(SO
4
)
2
·6H
2
O 0.2g
Bromocresol Purple 0.032
Agar 6.0g
pH 5.6 ± 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. Distribute into tubes.
Overlay with viscous parrafin. Autoclave for 15 min at 15 psi pressure–
121°C. Allow tubes to solidify in a vertical position.
Use: For the identification of members of Enterobacteriaceae on the
basis of lysine decarboxylase and hydrogen sulfide production.
© 2010 by Taylor and Francis Group, LLC
456 Cow Manure Agar
Cow Manure Agar
Composition per liter:
Cow manure 50.0g
Agar 15.0g
Preparation of Medium: Add cow manure to tap water and bring
volume to 1.0L. Gently heat and bring to boiling. Boil for 1 hr. Filter
through cheesecloth. Filter through Whatman filter paper. Bring volume to
1.0L with tap water. Add agar. Mix thoroughly. Gently heat and bring to

boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pres-
sure–121°C. Pour into sterile Petri dishes or leave in tubes.
Use: For the cultivation of Streptomyces species.
CP Medium
Composition per liter:
Peptone 2.5g
Starch 2.0g
NaNO
3
0.38g
Tris(hydroxymethyl)aminomethane buffer 0.25g
K
2
HPO
4
0.038g
MgSO
4
·7H
2
O 0.038g
CaCl
2
·2H
2
O 0.017g
NaCl 0.013g
TC vitamins minimal eagle, 100X 5.0mL
Solution 1 1.0mL
Solution 2 1.0mL

Solution 3 1.0mL
Solution 4 1.0mL
Vitamin B
12
solution 0.2mL
pH 8.7 ± 0.2 at 25°C
TC Vitamins Minimal Eagle 100X:
Composition
per liter:
Inositol 2.0mg
Choline chloride 1.0mg
Folic acid 1.0mg
Nicotinamide 1.0mg
Calcium pantothenate 1.0mg
Pyridoxal 1.0mg
Thiamine·HCl 1.0mg
Riboflavin 0.1mg
Preparation of TC Vitamins Minimal Eagle, 100X: Add com-
ponents to distilled/deionized water and bring volume to 1.0L. Mix
thoroughly. Filter sterilize.
Solution 1:
Composition
per 100.0mL:
EDTA 5.0g
KOH 3.1g
Preparation of Solution 1: Add components to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly.
Solution 2:
Composition
per liter:

FeSO
4
·7H
2
O 4.98g
Preparation of Solution 2: Add FeSO
4
·7H
2
O to distilled/deionized
water acidified with 1.0mL of H
2
SO
4
. Bring volume to 1.0L. Mix thor-
oughly.
Solution 3:
Composition
per 100.0mL:
H
3
BO
3
1.14g
Preparation of Solution 3: Add H
3
BO
3
to distilled/deionized water
and bring volume to 100.0mL. Mix thoroughly.

Solution 4:
Composition
per 100.0mL:
ZnSO
4
·7H
2
O 0.88g
MnCl
2
·4H
2
O 0.144g
MoO
3
0.071g
CoNO
3
·6H
2
O 0.049g
CuSO
4
·5H
2
O 0.016g
Preparation of Solution 4: Add components to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly.
Vitamin B
12

Solution
Composition
per 10.0mL:
Vitamin B
12
10.0mg
Preparation of Vitamin B
12
Solution: Add vitamin B
12
to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Filter sterilize.
Preparation of Medium: Add components, except for vitamin so-
lutions, to distilled/deionized water and bring volume to 995.0mL. Mix
thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15
psi pressure–121°C. Aseptically add vitamin solutions. Mix thorough-
ly.
Use: For the cultivation of Lysobacter species.
CP Medium for Coprothermobacter proteolyticus
Composition per 1010.0mL:
NaHCO
3
8.4g
Pancreatic digest of casein 2.0g
Yeast extract 2.0g
MgCl
2
·6H
2

O 1.0g
NH
4
Cl 1.0g
CaCl
2
·2H
2
O 0.4g
K
2
HPO
4
·3H
2
O 0.4g
Resazurin 0.5mg
Gelatin solution 100.0mL
Na
2
S·9H
2
O solution 10.0mL
Wolfe’s mineral solution 10.0mL
pH 7.0 ± 0.2 at 25°C
Gelatin Solution:
Composition
per 100.0mL:
Gelatin 3.0g
Preparation of Gelatin Solution: Add gelatin to distilled/deion-

ized water and bring volume to 100.0mL. Mix thoroughly. Sparge with
100% N
2
. Autoclave for 15 min at 15 psi pressure–121°C.
Na
2
S·9H
2
O Solution:
Composition
per 10.0mL:
Na
2
S·9H
2
O 0.5g
Preparation of Na
2
S·9H
2
O Solution: Add Na
2
S·9H
2
O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N
2
. Autoclave for 15 min at 15 psi pressure–121°C.
Before use, neutralize to pH 7.0 with sterile HCl.

Wolfe’s Mineral Solution:
Composition
per liter:
MgSO
4
·7H
2
O 3.0g
Nitrilotriacetic acid 1.5g
NaCl 1.0g
MnSO
4
·2H
2
O 0.5g
© 2010 by Taylor and Francis Group, LLC
CPC Agar Base with Cellobiose, Colistin, and Polymyxin B 457
CoCl
2
·6H
2
O 0.1g
ZnSO
4
·7H
2
O 0.1g
CaCl
2
·2H

2
O 0.1g
FeSO
4
·7H
2
O 0.1g
NiCl
2
·6H
2
O 0.025g
KAl(SO
4
)
2
·12H
2
O 0.02g
CuSO
4
·5H
2
O 0.01g
H
3
BO
3
0.01g
Na

2
MoO
4
·2H
2
O 0.01g
Na
2
SeO
3
·5H
2
O 0.3mg
Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with
KOH. Add remaining components. Add distilled/deionized water to
1.0L. Adjust pH to 6.8.
Preparation of Medium: Prepare medium anaerobically under
100% CO
2
. Add components, except gelatin solution, Na
2
S·9H
2
O so-
lution, and Wolfe’s mineral solution, to distilled/deionized water and
bring volume to 880.0mL. Mix thoroughly. Gently heat and bring to
boiling. Cool to room temperature while sparging with 100% CO
2
.

Sparge with 100% CO
2
for 20 min. Adjust pH to 7.0. Autoclave for
15 min at 15 psi pressure–121°C. Aseptically and anaerobically add
100.0mL of sterile gelatin solution, 10.0mL of sterile Na
2
S·9H
2
O so-
lution, and 10.0mL of sterile Wolfe’s mineral solution to each tube.
Mix thoroughly.
Use: For the cultivation of Coprothermobacter proteolyticus.
CP Medium for Thermobacteroides leptospartum
Composition per 1010.0mL:
NaHCO
3
8.4g
Pancreatic digest of casein 2.0g
Yeast extract 2.0g
MgCl
2
·6H
2
O 1.0g
NH
4
Cl 1.0g
CaCl
2
·2H

2
O 0.4g
K
2
HPO
4
·3H
2
O 0.4g
Resazurin 0.5mg
Glucose solution 100.0mL
Na
2
S·9H
2
O solution 10.0mL
Wolfe’s mineral solution 10.0mL
pH 7.0 ± 0.2 at 25°C
Glucose Solution:
Composition
per 100.0mL:
D-Glucose 5.0g
Preparation of Glucose Solution: Add glucose to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Sparge with
100% N
2
. Autoclave for 15 min at 15 psi pressure–121°C.
Na
2
S·9H

2
O Solution:
Composition
per 10.0mL:
Na
2
S·9H
2
O 0.5g
Preparation of Na
2
S·9H
2
O Solution: Add Na
2
S·9H
2
O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N
2
. Autoclave for 15 min at 15 psi pressure–121°C.
Before use, neutralize to pH 7.0 with sterile HCl.
Wolfe’s Mineral Solution:
Composition
per liter:
MgSO
4
·7H
2

O 3.0g
Nitrilotriacetic acid 1.5g
NaCl 1.0g
MnSO
4
·2H
2
O 0.5g
CoCl
2
·6H
2
O 0.1g
ZnSO
4
·7H
2
O 0.1g
CaCl
2
·2H
2
O 0.1g
FeSO
4
·7H
2
O 0.1g
NiCl
2

·6H
2
O 0.025g
KAl(SO
4
)
2
·12H
2
O 0.02g
CuSO
4
·5H
2
O 0.01g
H
3
BO
3
0.01g
Na
2
MoO
4
·2H
2
O 0.01g
Na
2
SeO

3
·5H
2
O 0.3mg
Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with
KOH. Add remaining components. Add distilled/deionized water to
1.0L. Adjust pH to 6.8.
Preparation of Medium: Prepare medium anaerobically under 100%
CO
2
. Add components, except glucose solution, Na
2
S·9H
2
O solution, and
Wolfe’s mineral solution, to distilled/deionized water and bring volume to
880.0mL. Mix thoroughly. Gently heat and bring to boiling. Cool to room
temperature while sparging with 100% CO
2
. Sparge with 100% CO
2
for
20 min. Adjust pH to 7.0. Autoclave for 15 min at 15 psi pressure–121°C.
Aseptically and anaerobically add 100.0mL of sterile glucose solution,
10.0mL of sterile Na
2
S·9H
2
O solution, and 10.0mL of sterile Wolfe’s min-

eral solution to each tube. Mix thoroughly.
Use: For the cultivation of Thermobacteroides leptospartum.
CPC Agar
See: Cellobiose Polymyxin Colistin Agar
CPC Agar Base
with Cellobiose, Colistin, and Polymyxin B
Composition per liter:
NaCl 20.0g
Agar 15.0g
Cellobiose 15.0g
Peptic digest of animal tissue 10.0g
Beef extract 5.0g
Bromthymol Blue 0.04g
Cresol Red 0.04g
Cellobiose colistin polymyxin B solution 100.0mL
pH 7.6 ± 0.2 at 25°C
Source: This medium, without cellobiose colistin polymyxin B solu-
tion, is available as a premixed powder from HiMedia.
Cellobiose Colistin Polymyxin B Solution:
Composition
per 100.0mL:
Cellobiose 15.0g
Colistin 1,360,000U
Polymyxin B 100,000U
Preparation of Cellobiose Colistin Polymyxin B Solution:
Add components to distilled/deionized water and bring volume to
100.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium: Add components, except cellobiose co-
listin polymyxin B solution, to tap water and bring volume to 1.0L.
Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min

at 15 psi pressure–121°C. Aseptically add 100.0mL of sterile cellobio-
se colistin polymyxin B solution to 900.0 mL of the cooled agar base.
Mix thoroughly. Pour into sterile Petri dishes. Use within 7 days.
© 2010 by Taylor and Francis Group, LLC
458 CPC HiVeg Agar Base with Cellobiose, Colistin, and Polymyxin B
Use: For the cultivation and identification of Vibrio species from
foods.
CPC HiVeg Agar Base
with Cellobiose, Colistin, and Polymyxin B
Composition per liter:
NaCl 20.0g
Agar 15.0g
Cellobiose 15.0g
Plant peptone 10.0g
Plant extract 5.0g
Bromthymol Blue 0.04g
Cresol Red 0.04g
Cellobiose colistin polymyxin B solution 100.0mL
pH 7.6 ± 0.2 at 25°C
Source: This medium, without cellobiose colistin polymyxin B solu-
tion, is available as a premixed powder from HiMedia.
Cellobiose Colistin Polymyxin B Solution:
Composition
per 100.0mL:
Cellobiose 15.0g
Colistin 1,360,000U
Polymyxin B 100,000U
Preparation of Cellobiose Colistin Polymyxin B Solution:
Add components to distilled/deionized water and bring volume to
100.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except cellobiose co-
listin polymyxin B solution, to tap water and bring volume to 1.0L.
Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min
at 15 psi pressure–121°C. Aseptically add 100.0mL of sterile cellobio-
se colistin polymyxin B solution to 900.0 mL of the cooled agar base.
Mix thoroughly. Pour into sterile Petri dishes. Use within 7 days.
Use: For the cultivation and identification of Vibrio species from
foods.
CPC Medium
Composition per liter:
Sucrose 30.0g
Peptone 2.0g
Casein hydrolysate 1.0g
K
2
HPO
4
·3H
2
O 1.0g
KCl 0.5g
MgSO
4
·7H
2
O 0.5g
FeSO
4
·7H
2

O 0.1g
pH 7.2 ± 0.2 at 25°C
Preparation of Medium: Add components to tap water and bring
volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Auto-
clave for 15 min at 15 psi pressure–121°C.
Use: For the cultivation of Actinoplanes species.
CR Agar
See: Congo Red Agar
Craig’s Medium
Composition per liter:
Casein acid hydrolysate 30.0g
Yeast extract 4.0g
K
2
HPO
4
0.5g
Glucose solution 20.0mL
pH 7.2 ± 0.2 at 25°C
Source: This medium is available from HiMedia.
Glucose Solution:
Composition
per 100.0mL:
Glucose 20.0g
Preparation of Glucose Solution: Add glucose to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Filter ster-
ilize.
Preparation of Medium: Add components, except glucose solu-
tion, to distilled/deionized water and bring volume to 980.0mL. Mix
thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to

50°C. Aseptically add glucose solution. Mix thoroughly. Aseptically
distribute into sterile tubes.
Use: For the enrichment and cultivation of Vibrio cholerae during test-
ing of enterotoxigenicity.
CRAMP Agar
See: Congo Red Acid Morpholinepropanesulfonic Acid
Pigmentation Agar
CRAMP HiVeg Agar Base
Composition per liter:
Agarose 14.0g
Morpholine propane sulfonic acid 8.4g
Tricine 1.8g
NaCl 2.9g
Galactose 2.0g
Plant acid hydrolysate 2.0g
Na
2
S
2
O
3
0.6g
NH
4
Cl 0.5g
K
2
HPO
4
0.24g

MgSO
4
0.0986g
Congo Red 0.005g
pH 5.3 ± 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. Adjust pH to 5.3. 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 Yersinia species with plasmids.
CREA
(Creatine Agar)
Composition per liter:
Sucrose 30.0g
Agar 15.0g
Creatine·H
2
O 3.0g
K
3
P0
4
·7H
2
O 1.6g
Bromcresol Purple 50.0mg
Minerals solution 10.0mL

Trace minerals solution 1.0mL
© 2010 by Taylor and Francis Group, LLC
Creatinine/NMH Medium 459
Minerals Solution:
Composition
per 100.0mL:
KCl 5.0g
MgSO
4
·7H
2
O 5.0g
FeSO
4
·7H
2
O 0.1g
Preparation of Minerals Solution: Add components to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly.
Trace Minerals Solution:
Composition
per 100.0mL:
ZnS0
4
·7H
2
O 1.0g
CuSO
4
·5H

2
O 0.5g
Preparation of Trace Minerals Solution: Add components to
distilled/deionized water and bring volume to 100.0mL. Mix thorough-
ly.
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 Penicillium species.
Creatinine Agar
Composition per liter:
Agar 15.0g
Na
2
HPO
4
·12H
2
O 9.0g
NaCl 5.0g
KH
2
PO
4
1.5g
Creatinine 1.0g
Meat extract 1.0g
Yeast extract 1.0g
MgSO

4
·7H
2
O 0.2g
MnCl
2
·4H
2
O 20.0mg
CaCl
2
1.2mg
Glucose solution 100.0mL
Glucose Solution:
Composition
per 100.0mL:
Glucose 5.0g
Preparation of Glucose Solution: Add glucose to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Filter ster-
ilize. Warm to 50°C.
Preparation of Medium: Add components, except glucose solu-
tion, to distilled/deionized water and bring volume to 900.0mL. Mix
thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at
15 psi pressure–121°C. Cool to 50°–55°C. Aseptically add 100.0mL of
sterile glucose solution. Mix thoroughly. Pour into sterile Petri dishes
or distribute into sterile tubes.
Use: For the cultivation and maintenance of Pseudomonas species
and other bacteria that can utilize creatinine.
Creatinine Medium
Composition per liter:

Creatinine 5.0g
Agar 2.0g
Fumaric acid 2.0g
K
2
HPO
4
2.0g
Yeast extract 1.0g
Salt solution 10.0mL
pH 6.8 ± 0.2 at 25°C
Salt Solution:
Composition
per liter:
MgSO
4
12.2g
FeSO
4
·7H
2
O 2.8g
MnSO
4
·H
2
O 1.7g
CaCl
2
·2H

2
O 0.76g
NaCl 0.6g
Na
2
MoO
4
·2H
2
O 0.1g
ZnSO
4
·7H
2
O 0.06g
HCl (0.1N solution) 1.0L
Preparation of Salt Solution: Dissolve salts in 1.0L of 0.1N HCl
solution. Mix thoroughly.
Preparation of Medium: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 6.8 with
NaOH or KOH. Gently heat until boiling. Distribute into tubes or flasks.
Autoclave for 15 min at 15 psi pressure–121°C.
Use: For the cultivation and maintenance of Pseudomonas species.
Creatinine Medium
(LMG 107)
Composition per liter:
Creatinine 5.0g
Fumaric acid 2.0g
K
2

HPO
4
2.0g
Yeast extract 1.0g
Salt solution 10.0mL
pH 6.8 ± 0.2 at 25°C
Salt Solution:
Composition
per liter:
MgSO
4
·7H
2
O 25.0g
FeSO
4
·7H
2
O 2.8g
MnSO
4
·H
2
O 1.7g
CaCl
2
·2H
2
O 0.76g
NaCl 0.6g

Na
2
MoO
4
·2H
2
O 0.1g
ZnSO
4
·7H
2
O 60.0mg
HCl (0.1M solution) 1.0L
Preparation of Salt Solution: Add components to distilled/deion-
ized water and bring volume to 1.0L. 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 and maintenance of Flavobacterium filamen-
tosum.
Creatinine/NMH Medium
Composition per 1100.0mL:
Yeast extract 5.0g
NaCl 1.0g
NaHCO
3
1.0g
MgSO
4
·7H

2
O 0.5g
Na
2
S·9H
2
O 0.5g
MnCl
2
·4H
2
O 0.06g
CaSO
4
·2H
2
O 0.05g
FeSO
4
·7H
2
O 0.01g
Na
2
SeO
3
·5H
2
O 26.0μg
Vitamin B

12
20.0μg
Resazurin 1.0mg
© 2010 by Taylor and Francis Group, LLC
460 CreDm1 Medium
Phosphate solution 100.0mL
Creatinine solution 100.0mL
Trace elements solution SL-4 10.0mL
Vitamin solution 10.0mL
L-Cysteine·HCl solution 10.0mL
Na
2
S·9H
2
O solution 10.0mL
pH 7.0 ± 0.2 at 25°C
Phosphate Solution:
Composition
per 100.0mL:
K
2
HPO
4
5.33g
KH
2
PO
4
2.64g
Preparation of Phosphate Solution: Add components to dis-

tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.
Sparge with 100% N
2
. Autoclave for 15 min at 15 psi pressure–121°C.
Creatinine Solution:
Composition
per 100.0mL:
Creatinine 5.5g
Preparation of Creatinine Solution: Add creatinine to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 100% N
2
. Filter sterilize.
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.
Preparation of Trace Elements Solution SL-4: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Vitamin Solution:
Composition
per liter:
Pyridoxine·HCl 10.0mg
Calcium
DL-pantothenate 5.0mg
Lipoic acid 5.0mg
Nicotinic acid 5.0mg
p-Aminobenzoic acid 5.0mg
Riboflavin 5.0mg
Thiamine·HCl 5.0mg
Biotin 2.0mg
Folic acid 2.0mg
Vitamin B
12
0.1mg
Preparation of Vitamin Solution: Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
L-Cysteine·HCl Solution:
Composition
per 10.0mL:
L-Cysteine·HCl 0.5g
Preparation of L-Cysteine·HCl Solution: Add L-cysteine·HCl to

distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Sparge with 100% N
2
. Autoclave for 15 min at 15 psi pressure–
121°C.
Na
2
S·9H
2
O Solution:
Composition
per 10.0mL:
Na
2
S·9H
2
O 0.5g
Preparation of Na
2
S·9H
2
O Solution: Add Na
2
S·9H
2
O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N
2
. Autoclave for 15 min at 15 psi pressure–121°C.

Preparation of Medium: Add components, except creatinine solu-
tion, phosphate solution, L-cysteine·HCl solution, and Na
2
S·9H
2
O so-
lution, to distilled/deionized water and bring volume to 880.0mL. Mix
thoroughly. Adjust pH to 7.0. Autoclave for 15 min at 15 psi pressure–
121°C. Aseptically and anaerobically add 100.0mL of sterile phos-
phate solution, 10.0mL of sterile
L-cysteine·HCl solution, and 10.0mL
of sterile Na
2
S·9H
2
O solution. Immediately prior to use, aseptically
and anaerobically add 100.0mL of sterile creatinine solution. Mix thor-
oughly. Aseptically and anaerobically distribute into tubes or bottles.
Use: For the cultivation and maintenance of Clostridium species.
CreDm1 Medium
Composition per 1002.0mL:
Solution A 980.0mL
Solution D (Vitamin solution) 10.0mL
Solution E 10.0mL
Solution B (Trace elements solution SL-10) 1.0mL
Solution C (Selentite-tungstate solution) 1.0mL
pH 6.7–6.9 at 25°C
Solution A:
Composition
per 980.0mL:

KH
2
PO
4
1.4g
NH
4
Cl 0.5g
MgCl
2
·6H
2
O 0.2g
CaCl
2
·2H
2
O 0.15g
Yeast extract 50.0mg
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 980.0mL. Mix thoroughly. Autoclave for 15
min at 15 psi pressure–121°C.
Solution B (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 Solution B (Trace Elements Solution SL-10):
Add FeCl
2
·4H
2
O 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. Autoclave for 15 min at 15 psi pressure–
121°C.
Solution C (Selenite-Tungstate Solution):
Composition
per liter:
NaOH 0.5g
Na
2
WO
4
·2H
2
O 4.0mg
Na
2
SeO
3

·5H
2
O 3.0mg
© 2010 by Taylor and Francis Group, LLC
Cryptoanaerobacter Medium 461
Preparation of Solution C (Selenite-Tungstate Solution):
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 D (Vitamin Solution):
Composition
per liter:
Pyridoxine·HCl 10.0mg
Calcium
DL-pantothenate 5.0mg
Lipoic acid 5.0mg
Nicotinic acid 5.0mg
p-Aminobenzoic acid 5.0mg
Riboflavin 5.0mg
Thiamine·HCl 5.0mg
Biotin 2.0mg
Folic acid 2.0mg
Vitamin B
12
0.1mg
Preparation of Solution D (Vitamin Solution): Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Filter sterilize.
Solution E:
Composition
per 10.0mL:

Disodium-DL-malate 1.6g
Preparation of Solution E: Add disodium-DL-malate to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Auto-
clave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Aseptically combine 980.0mL of sterile
solution A with 1.0mL of sterile solution B, 1.0mL of sterile solution
C, 10.0mL of sterile solution D, and 10.0mL of sterile solution E, in
that order. Mix thoroughly. Adjust pH to 6.7–6.9. Aseptically distribute
into sterile tubes or flasks.
Use: For the cultivation of Campylobacter species.
Crithidia Medium
Composition per liter:
Sucrose 15.0g
Pancreatic digest of casein 6.0g
Yeast extract 1.0g
Liver concentrate 0.1g
Hemin solution 5.0mL
pH 7.8 ± 0.2 at 25°C
Hemin Solution:
Composition
per 2.5mL:
Hemin 25.0mg
Triethanolamine (TEA) 2.5mL
Preparation of Hemin Solution: Add hemin to 2.5mL trietha-
nolamine. Mix thoroughly.
Preparation of Medium: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.8.
Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–
121°C. Slight hemin precipitate may occur.
Use: For the cultivation of Crithidia acanthocephali, Crithidia deanei,

Crithidia fasciculata, Crithidia harmosa, Crithidia hutneri, Crithidia
luciliae, Crithidia mellificae, Crithidia oncopelti, Crithidia species,
Herpetomonas samuelpessoai, Leptomonas pyrrhocoris, and Phyto-
monas davidi.
CRMOX Agar
See: Congo Red-Magnesium Oxalate Agar
Crossley Milk Medium
Composition per liter:
Skim milk powder 100.0g
Peptone 10.0g
Bromcresol Purple 0.1g
pH 5.8 ± 0.2 at 25°C
Source: This medium is available as a premixed powder from Oxoid
Unipath.
Preparation of Medium: Add components to a very small volume
of distilled/deionized water and mix to a paste. Gradually add more dis-
tilled/deionized water and bring volume to 1.0L. Distribute in 10.0mL
volumes into tubes. Autoclave for 5 min at 15 psi pressure–121°C.
Use: For the routine examination of canned food samples for anaero-
bic bacteria.
Cryptoanaerobacter Medium
(DSMZ Medium 1022)
Composition per liter:
Solution A 650.0mL
Clostridium sporogenes supernatant 350.0mL
pH 7.5–8.0 at 25°C
Solution A:
Composition
per 650.0mL:
Yeast extract 5.0g

NaHCO
3
4.0g
Casamino acids 1.0g
4-Hydroxybenzoic acid 0.45g
KH
2
PO
4
0.4g
NH
4
Cl 0.4g
Resazurin 0.5mg
Vitamin solution 10.0mL
Magnesium chloride solution 10.0mL
Calcium chloride solution 10.0mL
Trace element solution SL-10 1.0mL
Selenite/tungstate solution 1.0mL
Selenite/Tungstate Solution:
Composition
per liter:
NaOH 0.5g
Na
2
WO
4
·2H
2
O 4.0mg

Na
2
SeO
3
·5H
2
O 3.0mg
Preparation of Selenite/Tungstate Solution: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N
2
. Autoclave for 15 min at 15 psi pressure–121°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-
© 2010 by Taylor and Francis Group, LLC
462 CRYS Medium
oughly. Sparge with 100% N
2
. Autoclave for 15 min at 15 psi pressure–
121°C.
Vitamin Solution:
Composition
per liter:
Pyridoxine-HCl 10.0mg
Thiamine-HCl·2H
2
O 5.0mg
Riboflavin 5.0mg
Nicotinic acid 5.0mg
D-Ca-pantothenate 5.0mg
p-Aminobenzoic acid 5.0mg
Lipoic acid 5.0mg
Biotin 2.0mg
Folic acid 2.0mg
Vitamin B
12
0.1mg
Preparation of Vitamin Solution: Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Sparge
with 80% H
2

+ 20% CO
2
. Filter sterilize.
Magnesium Chloride Solution:
Composition
per 10.0mL:
MgCl
2
·6H
2
O 0.08g
Preparation of Magnesium Chloride Solution: Add
MgCl
2
·6H
2
O to distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Sparge with 100% N
2
. Autoclave for 15 min at 15 psi
pressure–121°C.
Calcium Chloride Solution:
Composition
per 10.0mL:
CaCl
2
·2H
2
O 0.06g
Preparation of Calcium Chloride Solution: Add CaCl

2
·2H
2
O
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N
2
. Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Solution A: Add components, except bicarbonate,
magnesium chloride and calcium chloride solution, to distilled/deion-
ized water and bring volume to 630.0mL. Mix thoroughly. Adjust pH
to 7.0–7.5. Gently heat and bring to boiling. Boil for 3 min. Cool while
sparging with 80% N
2
+ 20% CO
2
. Add the solid bicarbonate. Adjust
the pH to 7.8. Dispense under 80% N
2
+ 20% CO
2
gas atmosphere into
anaerobic vials. Autoclave for 15 min at 15 psi pressure–121°C. Asep-
tically and anoxically add magnesium chloride and calcium chloride
solutions. Adjust final pH of the medium to
pH 7.7. Note: It may be
necessary to add 10–20 mg sodium dithionite per liter (e.g., from 5%
(w/v) solution, freshly prepared under N
2
and filter sterilized), if the so-

lution is not completely reduced after inoculation.
Clostridium sporogenes Supernatant:
Composition
per liter:
Yeast extract 5.0g
NaHCO
3
4.0g
Casamino acids 1.0g
KH
2
PO
4
0.4g
NH
4
Cl 0.4g
Resazurin 0.5mg
Vitamin solution 10.0mL
Magnesium chloride solution 10.0mL
Calcium chloride solution 10.0mL
Na
2
S·9H
2
O solution 10.0mL
Trace element solution SL-10 1.0mL
Selenite-tungstate solution 1.0mL
Clostridium sporogenes Variable
Na

2
S·9H
2
O Solution:
Composition per 10.0mL:
Na
2
S·9H
2
O 0.3g
Preparation of Na
2
S·9H
2
O Solution: Add Na
2
S·9H
2
O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Autoclave under 100% N
2
for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
Preparation of Clostridium sporogenes Supernatant: Add
components, except bicarbonate, Na
2
S·9H
2
O solution, magnesium

chloride and calcium chloride solution, to distilled/deionized water and
bring volume to 980.0mL. Mix thoroughly. Adjust pH to 7.0–7.5. Gen-
tly heat and bring to boiling. Boil for 3 min. Cool while sparging with
80% N
2
+ 20% CO
2
. Add the solid bicarbonate. Adjust the pH to 7.8.
Dispense
under 80% N
2
+ 20% CO
2
gas atmosphere into anaerobic
bottles. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically
and anoxically add Na
2
S·9H
2
O solution, magnesium chloride and cal-
cium chloride solutions. Adjust final pH to
pH 7.0. Inoculate with
Clostridium sp. DSM 754. Incubate for 5 to 8 days at 37°C. Disrupt
cells of the grown culture by autoclaving for 20 min at 15 psi pressure–
121°C. Centrifuge autoclaved culture at 18,000g for 20 min. Discard
cell pellet. Store the supernatant in screw-capped bottles at −20°C. Be-
fore use sterilize the supernatant by autoclaving under 100% N
2
for 15
min at 15 psi pressure–121°C. Cool to room temperature under 100%

N
2
.
Preparation of Medium: Aseptically and anoxically combine
650.0mL of solution A with 350.0mL of Clostridum sporogenes super-
natant.
Use: For the cultivation of Cryptoanaerobacter spp.
Cryptococcus neoformans Screen Medium
See: CN Screen Medium
CRYS Medium
Composition per liter:
Stock extract 500.0mL
2× PP medium 500.0mL
Stock Extract:
Composition
per 500.0mL:
Cerophyll 5.0g
Brown rice 5.0g
Yeast extract 5.0g
Dried seaweed 5.0g
Preparation of Stock Extract: Add components to distilled/deion-
ized water and bring volume to 500.0mL. Mix thoroughly. Gently heat
and bring to boiling. Continue boiling for 5 min. Filter three times
through Whatman #1 filter paper while still hot. Cool to room temper-
ature. Adjust pH to 7.2. Bring volume to 500.0mL with distilled/deion-
ized water. Autoclave for 15 min at 15 psi pressure–121°C.
2× PP Medium:
Composition
per 500.0mL:
Proteose peptone 10.0g

Pancreatic digest of peptone 10.0g
Ribonucleic acid from Torula yeast 1.0g
Asolectin 0.2g
Artificial seawater 167.0mL
Vitamin solution 2.0mL
© 2010 by Taylor and Francis Group, LLC
Crystal Violet Lactose Agar 463
Artificial Seawater:
Composition
per 167.0mL:
Aqua-Marin sea salts 6.95g
Source: Aqua-Marin sea salts are available from Aquatrol, Inc., Ana-
heim, CA.
Preparation of Artificial Seawater: Add Aqua-Marin sea salts to
distilled/deionized water and bring volume to 167.0mL Mix thorough-
ly. Filter sterilize.
Vitamin Solution:
Composition
per 100.0mL:
Thiamine·HCl 150.0mg
Calcium
D-(+)-pantothenate 100.0mg
Folic acid 50.0mg
Nicotinamide 50.0mg
Pyridoxal·HCl 50.0mg
Riboflavin 50.0mg
DL-6-Thioctic acid 1.0mg
Biotin solution 10.0mL
Biotin Solution:
Composition

per 10.0mL:
Biotin 0.01mg
Preparation of Biotin Solution: Add biotin to 10.0mL of absolute
ethanol. Mix thoroughly.
Preparation of Vitamin Solution: Add components to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize. For long-term storage, preserve under nitrogen at −20°C.
Preparation of 2× PP Medium: Add asolectin to 200.0mL of dis-
tilled/deionized water. Gently heat to 80°C. Mix thoroughly. Add other
components, except artificial seawater and vitamin solution, to dis-
tilled/deionized water and bring volume to 331.0mL. Mix thoroughly.
Adjust pH to 7.2. Autoclave for 15 min at 15 psi pressure–121°C.
Aseptically add 167.0mL of sterile artificial seawater and 2.0mL of
sterile vitamin solution. Mix thoroughly.
Preparation of Medium: Aseptically combine 500.0mL of sterile
stock extract with 500.0mL of sterile 2× PP medium. Mix thoroughly.
Aseptically distribute into sterile tubes or flasks.
Use: For the cultivation of Pseudocohnilembus marinus.
Crystal Violet Agar
Composition per liter:
Agar 15.0g
Lactose 10.0g
Proteose peptone 5.0g
Beef extract 3.0g
Crystal Violet 3.3mg
pH 6.8 ± 0.1 at 25°C
Preparation of Medium: Add components to distilled/deionized wa-
ter and bring volume to 1.0L. Mix thoroughly. Gently heat until 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 differentiation of pathogenic staphylococci from non-
pathogenic staphylococci. Hemolytic and coagulating strains of Staph-
ylococcus aureus appear as purple or yellow colonies. Nonhemolytic
and noncoagulating strains of Staphylococcus species appear as white
colonies.
Crystal Violet Azide Esculin Agar
Composition per liter:
Agar 15.0g
Glucose 5.0g
NaCl 5.0g
Proteose peptone 5.0g
Pancreatic digest of casein 5.0g
Meat extract 3.0g
Esculin 1.0g
NaN
3
1.0g
Crystal Violet 0.1g
Bovine blood, citrated 100.0mL
pH 7.5 ± 0.2 at 25°C
Caution: Sodium azide is toxic. Azides also react with metals and
disposal must be highly diluted.
Preparation of Medium: Add components, except citrated bovine
blood, to distilled/deionized water and bring volume to 900.0mL. Mix
thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at
15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add sterile, cit-
rated bovine blood. Mix thoroughly. Pour into sterile Petri dishes or
distribute into sterile tubes.
Use: For the cultivation of Erysipelothrix rhusiopathiae.
Crystal Violet Esculin Agar

Composition per liter:
Agar 15.0g
Glucose 5.0g
NaCl 5.0g
Proteose peptone 5.0g
Pancreatic digest of casein 5.0g
Meat extract 3.0g
Esculin 1.0g
Crystal Violet 2.0mg
Blood, citrated 100.0mL
pH 7.5 ± 0.2 at 25°C
Preparation of Medium: Add components, except citrated blood,
to distilled/deionized water and bring volume to 900.0mL. Mix thor-
oughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi
pressure–121°C. Cool to 45°–50°C. Aseptically add citrated blood.
Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile
tubes.
Use: For the cultivation of Erysipelothrix rhusiopathiae.
Crystal Violet Lactose Agar
Composition per liter:
Agar 15.0g
Lactose 10.0g
Proteose peptone 5.0g
Beef extract 3.0g
Crystal Violet 3.3mg
pH 6.8 ± 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. Autoclave for 15 min at 15
psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.
© 2010 by Taylor and Francis Group, LLC
464 Crystal Violet Lactose Broth
Use: For the differentiation of pure cultures of pathogenic and non-
pathogenic staphylococci.
Crystal Violet Lactose Broth
Composition per liter:
Lactose 5.0g
Peptic digest of animal tissue 5.0g
K
2
HPO
4
5.0g
KH
2
PO
4
1.0g
Crystal Violet 1.43mg
pH 7.4 ± 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. Autoclave for 15 min at 15
psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.
Use: For the differentiation of pure cultures of pathogenic and non-
pathogenic staphylococci.

Crystal Violet Lactose HiVeg Agar
Composition per liter:
Agar 15.0g
Lactose 10.0g
Plant peptone No. 3 5.0g
Plant extract 3.0g
Crystal Violet 3.3mg
pH 6.8 ± 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. Autoclave for 15 min at 15
psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.
Use: For the differentiation of pure cultures of pathogenic and non-
pathogenic staphylococci.
Crystal Violet Pectate Medium
(CVP Medium)
Composition per liter:
Sodium polypectate 9.0g
Agar 2.0g
NaNO
3
1.0g
NaOH (1N solution) 4.5mL
CaCl
2
·H
2
O (10% solution) 3.0mL

Crystal Violet (0.075% solution) 1.0mL
Sodium lauryl sulfate (10% solution) 0.5mL
pH 7.2 ± 0.2 at 25°C
Preparation of Medium: In a preheated blender, add 500.0mL of
boiling distilled/deionized water and the components, except sodium
polypectate and sodium lauryl sulfate solution. Blend at high speed for
15 sec. Continue blending at low speed and slowly add 9.0g of sodium
polypectate. Pour the incomplete medium into a 2L flask and add
0.5mL of sodium lauryl sulfate solution. Mix thoroughly. Cap flask
with an aluminum foil seal rather than cotton. Autoclave for 25 min at
15 psi pressure–121°C. Pour medium quickly into sterile Petri dishes.
Allow plates to dry at 25°C for 48 hr before use.
Use: For the cultivation of pectinolytic microorganisms, such as
Erwinia species, from foods.
Crystal Violet Pectate Medium
(CVP Medium)
Composition per liter:
Sodium polypectate 18.0g
Agar 4.0g
NaNO
3
2.0g
CaCl
2
·2H
2
O 0.6g
NaOH 0.36g
Sodium lauryl sulfate 0.1g
Crystal Violet 1.5mg

pH 7.2 ± 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. Autoclave for 15 min at 15
psi pressure–121°C.
Use: For the cultivation of pectinolytic microorganisms, such as
Erwinia species, from foods.
Crystal Violet Tetrazolium Agar Base
Composition per liter:
Agar 15.0g
Casein enzymatic hydrolysate 5.0g
Yeast extract 2.5g
Glucose 1.0g
Crystal Violet 1.0mg
2,3,5-Triphenyltetrazolium chloride solution 10.0mL
pH 7.0 ± 0.2 at 25°C
Source: This medium, without triphenyltetrazolium chloride solution,
is available as a premixed powder from HiMedia.
Caution: Sodium azide is toxic. Azides also react with metals and
disposal must be highly diluted.
2,3,5-Triphenyltetrazolium Chloride Solution:
Composition
per 10.0mL:
2,3,5-Triphenyltetrazolium chloride 0.1g
Preparation of 2,3,5-Triphenyltetrazolium Chloride Solu-
tion:
Add 2,3,5-triphenyltetrazolium chloride to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except 2,3,5-triphe-
nyltetrazolium chloride solution, to distilled/deionized water and bring
volume to 990.0mL. Mix thoroughly. Gently heat and bring to boiling.
Autoclave for 10 min at 15 psi pressure–121°C. Cool to 45°–50°C.
Aseptically add 10.0mL 2,3,5-triphenyltetrazolium chloride solution.
Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile
tubes.
Use: For the detection of Gram-negative psychrotrophic bacteria caus-
ing food spoilage.
Crystal Violet Tetrazolium HiVeg Agar Base
Composition per liter:
Agar 15.0g
Plant hydrolysate 5.0g
Yeast extract 2.5g
Glucose 1.0g
© 2010 by Taylor and Francis Group, LLC