Handbook of Microbiological Media, Fourth Edition part 86 pdf

Hippea Medium 845
HiCrome™ UTI Agar, Modified
(UTI Agar, Modified HiCrome™)
Composition per liter:
Peptic digest of animal tissue 18.0g
Agar 15.0g
Chromogenic mixture 12.44g
Beef extract 4.0g
Casein enzymatic hydrolysate 4.0g
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 while
stirring and bring to boiling. Mix to completely dissolve components.
Do not autoclave. Cool to 50°C. Pour into sterile Petri dishes.
Use: A chromogenic medium used for detecting and identifying Enter-
obacteria, Proteus species, and other bacteria involved in urinary tract
infections.
HiFluoro™ Pseudomonas Agar Base
Composition per liter:
Pancreatic digest of gelatin 18.0g
Agar 15.0g
K
2
SO
4
10.0g
Fluorogenic mixture 2.05g
MnCl
2

1.4g
Cetrimide 0.3g
Glycerol 10.ml
pH 7.2 ± 0.2 at 25°C
Source: This medium, wihout glycerol, is available as a premixed
powder 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.
Use: For the selective isolation of Pseudomonas aeruginosa from clin-
ical and nonclinical specimens by the fluorogenic method.
High Plate Count Agar
Composition per liter:
Agar 15.0g
Peptic digest of animal tissue 3.0g
Casein, soluble 0.5g
K
2
HPO
4
0.2g
MgSO
4
·7H
2
O 0.05g
FeCl
3
·4H

2
O 1.0mg
pH 7.2 ± 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. Autoclave for 15 min at 15
psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.
Use: For obtaining higher colony counts by the spread plate, pour plate
or membrane filter technique.
High Salt Nutrient Agar
Composition per liter:
NaCl 30.0g
Agar 15.0g
Peptic digest of animal tissue 5.0g
Meat extract 5.0g
pH 8.5 ± 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. 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 salt-tolerant Vibrio species.
High Salt Peptone Yeast Extract Agar
Composition per liter:
NaCl 30.0g
Agar 15.0g
Peptic digest of animal tissue 10.0g
Yeast extract 6.0g
Meat extract 2.0g

Glucose 2.0g
L-Cysteine·HCl·H
2
O 0.3g
pH 7.5 ± 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. Autoclave for 15 min at 15
psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.
Use: For the confirmation of Vibrio species.
Hippea Medium
(DSMZ Medium 854)
Composition per 1010.0mL:
NaCl 25.0g
Sulfur, powdered 10.0g
Na-acetate 5.0g
MOPS [3-(N-morpholino) propane
sulfonic acid] 3.0g
Na
2
S·9H
2
O 0.5g
NH
4
Cl 0.33g
CaCl
2
·2H

2
O 0.33g
MgCl
2
·6H
2
O 0.33g
KCl 0.33g
KH
2
PO
4
0.33g
Yeast extract 0.1g
Resazurin 0.5mg
Trace elements solution 10.0mL
Vitamin solution 10.0mL
pH 6.1 ± 0.2 at 25°C
Trace Elements 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
CoSO
4
·7H
2
O 0.18g
ZnSO
4
·7H
2
O 0.18g
© 2010 by Taylor and Francis Group, LLC
846 Hippurate Hydrolysis Broth
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
H
3
BO
3
0.01g
Na
2
MoO
4
·4H
2
O 0.01g
CuSO
4
·5H
2
O 0.01g
Na
2
SeO
3
·5H
2

O 0.3mg
Preparation of Trace Elements Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 6.5 with KOH. Add remaining components. Add distilled/deionized
water to 1.0L. Mix thoroughly.
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.
Preparation of Medium: Add components, except vitamin solu-

tion, sulfur, and Na
2
S·9H
2
O, to distilled/deionized water and bring vol-
ume to 1.0L. Mix thoroughly. Adjust pH to 6.0. Sparge the medium
with 80% N
2
+ 20% CO
2
gas mixture for 30 min. Add Na
2
S·9H
2
O.
Mix thoroughly. Readjust the pH to 6.0–6.2. Dispense medium under
80% N
2
+ 20% CO
2
gas mixture into anaerobe tubes or bottles con-
taining 100.0mg sulfur powder per 10mL medium. Autoclave 20 min
at 110°C. Prior to use inject 0.1mL sterile vitamin solution per 10.0mL
medium.
Use: For the cultivation of Hippea maritima.
Hippurate Broth
See: Sodium Hippurate Broth
Hippurate Hydrolysis Broth
Composition per liter:
Heart infusion powder 10.0g

Peptic digest of animal tissue 10.0g
Sodium hippurate 10.0g
NaCl 5.0g
pH 7.4 ± 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. Autoclave for 15 min at 15
psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.
Use: For the detection of hippurate-hydrolyzing bacteria.
Hirschia Medium
Composition per liter:
Pancreatic digest of casein 5.0g
HEPES 4.0g
Yeast extract 2.0g
Artificial seawater 250.0mL
Glucose solution 100.0mL
pH 7.4 ± 0.2 at 25°C
Glucose Solution:
Composition
per 100.0mL:
Glucose 0.25g
Preparation of Glucose Solution: Add glucose to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Filter ster-
ilize.
Artificial Seawater:
Composition
per liter:
NaCl 27.5g
MgCl

2
·6H
2
O 5.38g
MgSO
4
·7H
2
O 6.78g
KCl 0.72g
NaHCO
3
0.2g
CaCL
2
·2H
2
O 1.4g
Preparation of Artificial Seawater: Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
Preparation of Medium: Add components, except glucose solu-
tion, to distilled/deionized water and bring volume to 1.0L. Mix thor-
oughly. Adjust pH to 7.4. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 25°C. Aseptically add 100.0mL of sterile glucose solu-
tion. Mix thoroughly. Aseptically distribute into sterile tubes or flasks.
Use: For the cultivation of Hirschia baltica.
Hi-Sensitivity Test Agar
Composition per liter:
Casein enzymic hydrolysate 11.0g
Agar 8.0g

NaCl 3.0g
Peptic digest of animal tissue 3.0g
Glucose 2.0g
Na
2
HPO
4
2.0g
Sodium acetate 1.0g
Starch, soluble 1.0g
Magnesium glycerophosphate 0.2g
Calcium gluconate 0.1g
L-Cystine hydrochloride 0.02g
L-Tryptophan 0.02g
Adenine 0.01g
Guanine 0.01g
Uracil 0.01g
Xanthine 0.01g
Calcium pantothenate 3.0mg
Biotin 3.0mg
Nicotinamide 3.0mg
Pyridoxine hydrochloride 3.0mg
Manganese chloride 2.0mg
ZnSO
4
1.0mg
CoSO
4
1.0mg
CuSO

4
1.0mg
Cyanocobalamin 1.0mg
FeSO
4
1.0mg
Menadione 1.0mg
Thiamine hydrochloride 0.04mg
pH 7.2 ± 0.2 at 25°C
© 2010 by Taylor and Francis Group, LLC
Hi-Sensitivity Test HiVeg Broth 847
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.
Use: For antimicrobial susceptibility tests.
Hi-Sensitivity Test Broth
Composition per liter:
Casein enzymic hydrolysate 11.0g
NaCl 3.0g
Peptic digest of animal tissue 3.0g
Glucose 2.0g
Na
2
HPO
4
2.0g
Sodium acetate 1.0g

Starch, soluble 1.0g
Magnesium glycerophosphate 0.2g
Calcium gluconate 0.1g
L-Cystine hydrochloride 0.02g
L-Tryptophan 0.02g
Adenine 0.01g
Guanine 0.01g
Uracil 0.01g
Xanthine 0.01g
Calcium pantothenate 3.0mg
Biotin 3.0mg
Nicotinamide 3.0mg
Pyridoxine hydrochloride 3.0mg
Manganese chloride 2.0mg
ZnSO
4
1.0mg
CoSO
4
1.0mg
CuSO
4
1.0mg
Cyanocobalamin 1.0mg
FeSO
4
1.0mg
Menadione 1.0mg
Thiamine hydrochloride 0.04mg
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 antimicrobial susceptibility testing.
Hi-Sensitivity Test HiVeg Agar
Composition per liter:
Plant hydrolysate 11.0g
Agar 8.0g
NaCl 3.0g
Plant peptone 3.0g
Glucose 2.0g
Na
2
HPO
4
2.0g
Sodium acetate 1.0g
Starch, soluble 1.0g
Magnesium glycerophosphate 0.2g
Calcium gluconate 0.1g
L-Cystine hydrochloride 0.02g
L-Tryptophan 0.02g
Adenine 0.01g
Guanine 0.01g
Uracil 0.01g
Xanthine 0.01g
Calcium pantothenate 3.0mg

Biotin 3.0mg
Nicotinamide 3.0mg
Pyridoxine hydrochloride 3.0mg
Manganese chloride 2.0mg
ZnSO
4
1.0mg
CoSO
4
1.0mg
CuSO
4
1.0mg
Cyanocobalamin 1.0mg
FeSO
4
1.0mg
Menadione 1.0mg
Thiamine hydrochloride 0.04mg
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.
Use: For antimicrobial susceptibility tests.
Hi-Sensitivity Test HiVeg Broth
Composition per liter:
Plant hydrolysate 11.0g

NaCl 3.0g
Plant peptone 3.0g
Glucose 2.0g
Na
2
HPO
4
2.0g
Sodium acetate 1.0g
Starch, soluble 1.0g
Magnesium glycerophosphate 0.2g
Calcium gluconate 0.1g
L-Cystine hydrochloride 0.02g
L-Tryptophan 0.02g
Adenine 0.01g
Guanine 0.01g
Uracil 0.01g
Xanthine 0.01g
Calcium pantothenate 3.0mg
Biotin 3.0mg
Nicotinamide 3.0mg
Pyridoxine hydrochloride 3.0mg
Manganese chloride 2.0mg
ZnSO
4
1.0mg
CoSO
4
1.0mg
CuSO

4
1.0mg
Cyanocobalamin 1.0mg
FeSO
4
1.0mg
Menadione 1.0mg
Thiamine hydrochloride 0.04mg
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
© 2010 by Taylor and Francis Group, LLC
848 Hisitest Agar
to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15
psi pressure–121°C.
Use: For antimicrobial susceptibility testing.
Hisitest Agar
Composition per liter:
Casein enzymic hydrolysate 11.0g
Agar 8.0g
Buffer salt 3.3g
Peptic digest of animal tissue 3.0g
NaCl 3.0g
Glucose 2.0g
Starch 1.0g
Nucleoside basis 0.02g
Thiamine 0.02mg
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 determination of antibiotic susceptibility of fastidious micro-
organisms.
Histidans Agar
Composition per liter:
Agar 20.0g
Glucose 10.0g
Yeast extract 10.0g
Na
2
HPO
4
0.95g
KH
2
PO
4
0.91g
MgSO
4
·7H
2
O 0.5g
pH 7.0 ± 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 of Streptomyces species.
Histoplasma capsulatum Agar
Composition per liter:
Agar 12.5g
Glucose 10.0g
Citric acid 10.0g
Potato starch 2.0g
α-Ketoglutaric acid 1.0g
L-Cystine·HCl·H
2
O 1.0g
Glutathione, reduced 0.5g
L-Asparagine 0.1g
L-Tryptophan 0.02g
Solution 1 250.0mL
Solution 3 40.0mL
Solution 2 10.0mL
Solution 4 10.0mL
Solution 8 10.0mL
Solution 5 1.0mL
Solution 6 0.1mL
Solution 7 0.1mL
pH 6.5 ± 0.2 at 25°C
Solution 1:
Composition
per liter:
KH

2
PO
4
8.0g
(NH
4
)
2
SO
4
8.0g
MgSO
4
·7H
2
O 0.86g
CaCl
2
, anhydrous 0.08g
ZnSO
4
·7H
2
O 0.05g
Preparation of Solution 1: Add components to distilled/deionized
water and bring volume to 500.0mL. Mix thoroughly. Bring volume to
1.0L with distilled/deionized water. Store at 5°C.
Solution 2:
Composition
per liter:

FeSO
4
·7H
2
O 5.7g
MnCl
2
·6H
2
O 0.8g
NaMoO
4
·2H
2
O 0.15g
HCl, concentrated 1.0mL
Preparation of Solution 2: Add 1.0mL of concentrated HCl to
100.0mL of distilled water in a 1.0L volumetric flask. Dissolve each
component completely in the sequence given. Bring volume to 1.0L
with distilled/deionized water. Store at 5°C. Discard if red color or red
precipitate appears.
Solution 3:
Composition
per 100.0mL:
Casein, acid-hydrolyzed, vitamin-free 10.0g
Preparation of Solution 3: Add casein to distilled/deionized water
and bring volume to 100.0mL.
Solution 4:
Composition
per liter:

Calcium pantothenate 0.2g
Inositol 0.2g
Riboflavin 0.2g
Thiamine·HCl 0.2g
Nicotinamide 0.1g
Biotin 0.01g
Preparation of Solution 4: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Store at −20°C.
Solution 5:
Composition
per 100.0mL:
Hemin 0.2g
NH
4
OH, concentrated 0.3mL
Preparation of Solution 5: Add hemin to approximately 30.0mL
of distilled/deionized water. Add NH
4
OH. Mix thoroughly until dis-
solved. Bring volume to 100.0mL with distilled/deionized water. Store
at 5° C.
Solution 6:
Composition
per 10.0mL:
DL-Thioctic acid 0.01g
Ethanol (95% solution) 10.0mL
Preparation of Solution 6: Add DL-thioctic acid to 10.0mL of eth-
anol. Mix thoroughly. Store at −20°C.
Solution 7:
Composition

per 10.0mL:
Coenzyme A 0.01g
Na
2
S·5H
2
O (0.05% solution) 0.2mL
© 2010 by Taylor and Francis Group, LLC
Histoplasma capsulatum Agar 849
Preparation of Solution 7: Prepare Na
2
S·5H
2
O solution in freshly
boiled distilled/deionized water. Add coenzyme A to 9.8mL of dis-
tilled/deionized water. Mix thoroughly. Add freshly prepared
Na
2
S·5H
2
O solution. Mix thoroughly. Store the solution at −20°C.
Solution 8:
Composition
per 100.0mL:
Oleic acid 0.1g
Preparation of Solution 8: Add oleic acid to 50.0mL of distilled/
deionized water. Adjust pH to 9.0 with NaOH. Gently heat until dis-
solved. Bring volume to 100.0mL with distilled/deionized water. Store
at 5°C.
Preparation of Medium: Add components—except agar, potato

starch, and solution 8—to distilled/deionized water and bring volume
to 400.0mL. Mix thoroughly. Adjust pH to 6.5 with 20% KOH solu-
tion. Filter sterilize. In a separate flask, add potato starch to 50.0mL of
distilled/deionized water. Add the starch solution to 450.0mL of boil-
ing distilled/deionized water. Add 10.0mL of solution 8 and the agar.
Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool
to 70°C. Aseptically combine the two sterile solutions. Pour into sterile
Petri dishes or distribute into sterile tubes.
Use: For the cultivation and maintenance of Histoplasma capsulatum
in the yeast phase. For the cultivation of Histoplasma duboisii, Blasto-
myces dermatitidis, and Sprotrichum schenckii.
Histoplasma capsulatum Agar
Composition per liter:
Agar 15.0g
Glucose 10.0g
Potato starch 2.0g
α-Ketoglutaric acid 1.0g
L-Cystine·HCl·H
2
O 1.0g
Glutathione, reduced 0.5g
L-Asparagine 0.1g
L-Tryptophan 0.02g
Solution 1 250.0mL
Solution 3 40.0mL
Solution 2 10.0mL
Solution 4 10.0mL
Solution 8 10.0mL
Solution 5 1.0mL
Solution 6 0.1mL

Solution 7 0.1mL
pH 6.5 ± 0.2 at 25°C
Solution 1:
Composition
per liter:
KH
2
PO
4
8.0g
(NH
4
)
2
SO
4
8.0g
MgSO
4
·7H
2
O 0.86g
CaCl
2
, anhydrous 0.08g
Preparation of Solution 1: Add components to distilled/deionized
water and bring volume to 500.0mL. Mix thoroughly. Bring volume to
1.0L with distilled/deionized water. Store at 5°C.
Solution 2:
Composition

per liter:
FeSO
4
·7H
2
O 5.7g
MnCl
2
·6H
2
O 0.8g
NaMoO
4
·2H
2
O 0.15g
HCl, concentrated 1.0mL
Preparation of Solution 2: Add the 1.0mL of concentrated HCl to
100.0mL of distilled water in a 1.0L volumetric flask. Dissolve each
component completely in the sequence given. Bring volume to 1.0L
with distilled/deionized water. Store at 5°C. Discard if red color or red
precipitate appears.
Solution 3:
Composition
per 100.0mL:
Casein, acid-hydrolyzed, vitamin-free 10.0g
Preparation of Solution 3: Add casein to distilled/deionized water
and bring volume to 100.0mL. Do not use enzymatically digested ca-
sein.
Solution 4:

Composition
per liter:
Calcium pantothenate 0.2g
Inositol 0.2g
Riboflavin 0.2g
Thiamine·HCl 0.2g
Nicotinamide 0.1g
Biotin 0.01g
Preparation of Solution 4: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Store at −20°C.
Solution 5:
Composition
per 100.0mL:
Hemin 0.2g
NH
4
OH, concentrated 0.3mL
Preparation of Solution 5: Add hemin to approximately 30.0mL
of distilled/deionized water. Add NH
4
OH. Mix thoroughly until dis-
solved. Bring volume to 100.0mL with distilled/deionized water. Store
at 5°C.
Solution 6:
Composition
per 10.0mL:
DL-Thioctic acid 0.01g
Ethanol (95% solution) 10.0mL
Preparation of Solution 6: Add DL-thioctic acid to 10.0mL of
ethanol. Mix thoroughly. Store solution at −20°C.

Solution 7:
Composition
per 10.0mL:
Coenzyme A 0.01g
Na
2
S·5H
2
O (0.05% solution) 0.2mL
Preparation of Solution 7: Prepare Na
2
S·5H
2
O solution in freshly
boiled distilled/deionized water. Add coenzyme A to 9.8mL of dis-
tilled/deionized water. Mix thoroughly. Add freshly prepared
Na
2
S·5H
2
O solution. Mix thoroughly. Store the solution at −20°C.
Solution 8:
Composition
per 100.0mL:
Oleic acid 0.1g
Preparation of Solution 8: Add oleic acid to 50.0mL of distilled/
deionized water. Adjust pH to 9.0 with NaOH. Gently heat until dis-
solved. Bring volume to 100.0mL with distilled/deionized water. Store
at 5°C.
Preparation of Medium: Add components—except agar, potato

starch, and solution 8—to distilled/deionized water and bring volume
to 400.0mL. Mix thoroughly. Adjust pH to 6.5 with 20% KOH solu-
tion. Filter sterilize. In a separate flask, add potato starch to 50.0mL of
distilled/deionized water. Add the starch solution to 450.0mL of boil-
ing distilled/deionized water. Add 10.0mL of solution 8 and the agar.
Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool
© 2010 by Taylor and Francis Group, LLC
850 Histoplasma capsulatum Broth
to 70°C. Aseptically combine the two sterile solutions. Pour into sterile
Petri dishes or distribute into sterile tubes.
Use: For the cultivation and maintenance of Histoplasma capsulatum
in the mycelial phase.
Histoplasma capsulatum Broth
Composition per liter:
Glucose 10.0g
Citric acid 10.0g
α-Ketoglutaric acid 1.0g
L-Cystine·HCl·H
2
O 1.0g
Potato starch 0.5g
Glutathione, reduced 0.5g
L-Asparagine 0.1g
L-Tryptophan 0.02g
Solution 1 250.0mL
Solution 3 40.0mL
Solution 2 10.0mL
Solution 4 10.0mL
Solution 5 1.0mL
Solution 8 1.0mL

Solution 6 0.1mL
Solution 7 0.1mL
pH 6.5 ± 0.2 at 25°C
Solution 1:
Composition
per liter:
KH
2
PO
4
8.0g
(NH
4
)
2
SO
4
8.0g
MgSO
4
·7H
2
O 0.86g
CaCl
2
, anhydrous 0.08g
ZnSO
4
·7H
2

O 0.05g
Preparation of Solution 1: Add components to distilled/deionized
water and bring volume to 500.0mL. Mix thoroughly. Bring volume to
1.0L with distilled/deionized water. Store at 5°C.
Solution 2:
Composition
per liter:
FeSO
4
·7H
2
O 5.7g
MnCl
2
·6H
2
O 0.8g
NaMoO
4
·2H
2
O 0.15g
HCl, concentrated 1.0mL
Preparation of Solution 2: Add 1.0mL of concentrated HCl to
100.0mL of distilled water in a 1.0L volumetric flask. Dissolve each
component completely in the sequence given. Bring volume to 1.0L
with distilled/deionized water. Store at 5°C. Discard if red color or red
precipitate appears.
Solution 3:
Composition

per 100.0mL:
Casein, acid-hydrolyzed, vitamin-free 10.0g
Preparation of Solution 3: Add casein to distilled/deionized water
and bring volume to 100.0mL. Do not use enzymatically digested ca-
sein.
Solution 4:
Composition
per liter:
Calcium pantothenate 0.2g
Inositol 0.2g
Riboflavin 0.2g
Thiamine·HCl 0.2g
Nicotinamide 0.1g
Biotin 0.01g
Preparation of Solution 4: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Store at −20°C.
Solution 5:
Composition
per 100.0mL:
Hemin 0.2g
NH
4
OH, concentrated 0.3mL
Preparation of Solution 5: Add hemin to approximately 30.0mL
of distilled/deionized water. Add NH
4
OH. Mix thoroughly until dis-
solved. Bring volume to 100.0mL with distilled/deionized water. Store
at 5°C.
Solution 6:

Composition
per 10.0mL:
DL-Thioctic acid 0.01g
Ethanol (95% solution) 10.0mL
Preparation of Solution 6: Add DL-thioctic acid to 10.0mL of eth-
anol. Mix thoroughly. Store solution at −20°C.
Solution 7:
Composition
per 10.0mL:
Coenzyme A 0.01g
Na
2
S·5H
2
O (0.05% solution) 0.2mL
Preparation of Solution 7: Prepare Na
2
S·5H
2
O solution in freshly
boiled distilled/deionized water. Add coenzyme A to 9.8mL of dis-
tilled/deionized water. Mix thoroughly. Add freshly prepared
Na
2
S·5H
2
O solution. Mix thoroughly. Store the solution at −20°C.
Solution 8:
Composition
per 100.0mL:

Oleic acid 0.1g
Preparation of Solution 8: Add oleic acid to 50.0mL of distilled/de-
ionized water. Adjust pH to 9.0 with NaOH. Gently heat until dissolved.
Bring volume to 100.0mL with distilled/deionized water. Store at 5°C.
Preparation of Medium: Add components—except potato starch
and solution 8—to distilled/deionized water and bring volume to
400.0mL. Mix thoroughly. Adjust pH to 6.5 with 20% KOH solution.
Filter sterilize. In a separate flask, add potato starch to 50.0mL of dis-
tilled/deionized water. Add the starch solution to 450.0mL of boiling
distilled/deionized water. Add 1.0mL of solution 8. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 70°C. Asepti-
cally combine the two sterile solutions. Pour into sterile Petri dishes or
distribute into sterile tubes.
Use: For the cultivation of Histoplasma capsulatum in the yeast phase.
For the cultivation of Histoplasma duboisii, Blastomyces dermatitidis,
and Sprotrichum schenckii.
HiVeg Hydrolysate Agar with 2.5% Agar
Composition per liter:
Agar 25.0g
Plant hydrolysate 5.0g
Plant peptone 5.0g
NaCl 5.0g
Na
2
HPO
4
2.5g
Plant infusion 1.5g
Yeast autolysate 1.5g
Glycerol 22.0mL

pH 7.8 ± 0.2 at 25°C
© 2010 by Taylor and Francis Group, LLC
HNS Agar 851
Source: This medium, without glycerol, is available as a premixed
powder 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. Autoclave for 15 min at 15 psi pressure–121°C. Mix thor-
oughly. Pour into sterile Petri dishes or distribute into sterile tubes.
Use: For the cultivation of Vibrio cholerae. For the production of
cholera vaccine.
HiVeg Magnesium Broth
Composition per liter:
Plant hydrolysate 10.0g
NaCl 5.0g
MgSO
4
0.94g
pH 7.0 ± 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. Autoclave for 15 min at 15 psi pressure–121°C. Mix thor-
oughly.
Use: For the cultivation of recombinant strains of Escherichia coli.
HiVeg Peptone Water
Composition per liter:
Plant peptone 10.0g
NaCl 5.0g

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. Autoclave for 15 min at 15 psi pressure–121°C. Mix thor-
oughly.
Use: For the cultivation of various bacteria.
HL Agar
Composition per plate:
Columbia agar base 10.0mL
Columbia blood top agar 5.0mL
pH 7.3 ± 0.2 at 25°C
Columbia Agar Base:
Composition per liter:
Agar 13.5g
Pancreatic digest of casein 12.0g
NaCl 5.0g
Peptic digest of animal tissue 5.0g
Beef extract 3.0g
Yeast extract 3.0g
Cornstarch 1.0g
Preparation of Columbia Agar Base: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gen-
tly heat until boiling. Autoclave for 15 min at 15 psi pressure–121°C.
Cool to 45°–50°C.
Columbia Blood Top Agar:
Composition per liter:
Agar 13.5g
Pancreatic digest of casein 12.0g

NaCl 5.0g
Peptic digest of animal tissue 5.0g
Beef extract 3.0g
Yeast extract 3.0g
Cornstarch 1.0g
Horse blood, defibrinated 50.0mL
Preparation of Columbia Blood Top Agar: Add components,
except horse blood, to distilled/deionized water and bring volume to
950.0mL. Mix thoroughly. Gently heat until boiling. Autoclave for 15
min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add ster-
ile horse blood. Mix thoroughly.
Preparation of Medium: Pour cooled, sterile Columbia agar base
into sterile Petri dishes in 10.0mL volumes. Allow agar to solidify.
Pour 5.0mL of cooled, sterile Columbia blood top agar over Columbia
agar base that has solidified but is still warm.
Use: For the cultivation of Listeria monocytogenes.
HM Medium
Composition per liter:
NaCl 81.0g
Yeast extract 10.0g
MgSO
4
9.6g
MgCl
2
7.0g
Proteose peptone No. 3 5.0g
KCl 2.0g
Glucose 1.0g
CaCl

2
0.36g
NaHCO
3
60.0mg
NaBr 26.0mg
pH 7.1 ± 0.2 at 25°C
Preparation of Medium: Add components to distilled/deionized wa-
ter and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.1. Distribute
into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.
Use: For the cultivation of Salinicoccus roseus and Salinicoccus hispani-
cus.
HNS Agar
(ATCC Medium 923)
Composition per liter:
Agar 15.0g
NaCl 9.6g
Heart infusion broth 990.0mL
Horse serum 10.0mL
pH 7.4 ± 0.2 at 25°C
Heart Infusion Broth:
Composition
per 900.0mL:
Beef heart, infusion from 500.0g
Tryptose 10.0g
NaCl 5.0g
Preparation of Heart Infusion Broth: Add agar and NaCl to
990.0mL heart infusion broth. Mix thoroughly. Autoclave for 15 min
at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add 10.0mL
sterile horse serum. Mix thoroughly. Pour into sterile Petri dishes or

distribute into sterile tubes.
© 2010 by Taylor and Francis Group, LLC
852 HNW Medium
Use: For the cultivation and maintenance of Corynebacterium species.
HNW Medium
(DSMZ Medium 997)
Composition per liter:
DMJ synthetic seawater 1.0L
Vitamin solution 10.0mL
NaHCO
3
solution 10.0mL
NaNO
3
solution 10.0mL
Na
2
S·9H
2
O solution 10.0mL
Tungstate solution 10.0mL
pH 7.2 ± 0.2 at 25°C
Tungstate Solution:
Composition
per 10.0mL:
Na
2
WO
4
·2H

2
O 0.1mg
Preparation of Tungstate Solution: Add Na
2
WO
4
·2H
2
O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
NaHCO
3
Solution:
Composition per 10.0mL:
NaHCO
3
1.0g
Preparation of NaHCO
3
Solution: Add NaHCO
3
to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 20% CO
2
+ 80% H
2
. Filter sterilize.
NaNO
3

Solution:
Composition per 10.0mL:
NaNO
3
1.0g
Preparation of NaNO
3
Solution: Add NaNO
3
to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with
100% N
2
. Filter sterilize.
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.
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.
Autoclave under 100% N

2
for 15 min at 15 psi pressure–121°C. Cool
to room temperature. Adjust pH to 7.5.
DMJ Synthetic Seawater:
Composition
per liter:
NaCl 30.0g
MgCl
2
·6H
2
O 4.18g
MgSO
4
·7H
2
O 3.4g
KCl 0.33g
NH
4
Cl 0.25g
K
2
HPO
4
0.14g
CaCl
2
·2H
2

O 0.14g
Fe(NH
4
)
2
(SO
4
)
2
·6H
2
O 0.01g
NiCl
2
·6H
2
O 0.5mg
Na
2
SeO
3
·5H
2
O 0.5mg
Trace elements solution SL-10 10.0mL
Trace Elements Solution SL-10:
Composition
per liter:
MgSO
4

·7H
2
O 3.0g
Nitrilotriacetic acid 1.5g
NaCl 1.0g
MnSO
4
·2H
2
O 0.5g
CoSO
4
·7H
2
O 0.18g
ZnSO
4
·7H
2
O 0.18g
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
H
3
BO
3
0.01g
Na
2
MoO
4
·4H
2
O 0.01g
CuSO
4
·5H
2
O 0.01g
Na

2
SeO
3
·5H
2
O 0.3mg
Preparation of Trace Elements Solution SL-10: Add nitrilotri-
acetic acid to 500.0mL of distilled/deionized water. Dissolve by adjust-
ing pH to 6.5 with KOH. Add remaining components. Add distilled/
deionized water to 1.0L. Mix thoroughly. Adjust pH to 7.0.
Preparation of DMJ Synthetic Seawater: Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temper-
ature.
Preparation of Medium: Aseptically add 10.0mL each of vitamin
solution, NaHCO
3
solution, NaNO
3
solution, Na
2
S·9H
2
O solution, and
tungstate solution to 1.0L sterile DMJ synthetic seawater. Mix thor-
oughly. Distribute into tubes. Tightly seal the tubes with butyl rubber
stoppers under a gas phase of 80% H
2
+ 20% CO
2

(300 kPa).
Use: For the cultivation of Persephonella hydrogeniphila and Hydro-
genivirga caldilitoris.
Hofer’s Alkaline Medium
Composition per liter:
Agar 15.0g
Mannitol 10.0g
Yeast extract 1.0g
K
2
HPO
4
0.5g
MgSO4 0.2g
NaCl 0.1g
Thymol Blue 0.016g
pH 11.0 ± 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. Autoclave for 15 min at 15
psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.
Use: For the selective isolation of Agrobacterium spp. from soil.
© 2010 by Taylor and Francis Group, LLC
Horikoshi-1 Medium with 10% Sodium Chloride 853
Hohn’s Medium, Modified
See: Steenken and Smith Agar
HO-LE Trace Elements Solution
Composition per liter:
H

3
BO
3
2.85g
MnCl
2
·4H
2
O 1.8g
Sodium tartrate 1.77g
FeSO
4
1.36g
CoCl
2
·6H
2
O 0.04g
CuCl
2
·2H
2
O 0.026g
Na
2
MoO
4
·2H
2
O 0.025g

ZnCl
2
0.021g
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 use as an enrichment to other media that require trace miner-
als.
Hominis Agar
See: H Agar
Hominis Broth
See: H Broth
Horie Arabinose Ethyl Violet Broth
(HAEB)
Composition per liter:
NaCl 30.0g
Peptone 5.0g
Beef extract 3.0g
Bromthymol Blue 0.03g
Ethyl Violet 1.0mg
Arabinose solution 100.0mL
pH 9.0 ± 0.2 at 25°C
Arabinose Solution:
Composition
per 100.0mL:
Arabinose 5.0g
Preparation of Arabinose Solution: Add arabinose to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize.
Preparation of Medium: Add components, except arabinose solu-

tion, to distilled/deionized water and bring volume to 900.0mL. Mix
thoroughly. Gently heat and bring to boiling. Adjust pH to 9.0. Auto-
clave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Asepti-
cally add sterile arabinose solution. Mix thoroughly. Aseptically
distribute into sterile tubes or flasks.
Use: For the cultivation of Vibrio species from foods.
Horikoshi Alkaline Medium
(DSMZ Medium 940)
Composition per liter:
Agar 15.0g
D-glucose 10.0g
Peptone 5.0g
Yeast extract 5.0g
Na
2
CO
3
5.0g
KH
2
PO
4
1.0g
MgSO
4
·7H
2
O 0.2g
pH 9.0 ± 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 Pannonibacter phragmite-
tus.
Horikoshi-1 Medium
(DSMZ Medium 1081)
Composition per liter:
Agar 15.0g
Glucose 10.0g
Polypeptone 5.0g
Yeast extract 5.0g
KH
2
PO
4
1.0g
K
2
HPO
4
1.0g
MgSO
4
·7H
2
O 0.2g
NaCO
3
solution 100.0mL

pH 10.0 ± 0.2 at 25°C
NaCO
3
Solution:
Composition per 100.0mL:
NaCO
3
10.0g
Preparation of NaCO
3
Solution: Add NaCO
3
to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Filter ster-
ilize.
Preparation of Medium: Add components, except NaCO
3
solu-
tion, to double distilled/deionized water and bring volume to 900.0mL.
Mix thoroughly. Adjust pH to 10.0. Gently heat while stirring and bring
to boiling. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 50°C. Aseptically add 100.0mL NaCO
3
solution. Ad-
just pH to 10.0. Pour into Petri dishes or aseptically distribute into
tubes.
Use: For the cultivation of “Streptomyces sannurensis” and Salinicoc-
cus alkaliphilus.
Horikoshi-1 Medium with 10% Sodium Chloride
(DSMZ Medium 1081a)

Composition per liter:
NaCl 100.0g
Agar 15.0g
Glucose 10.0g
Polypeptone 5.0g
Yeast extract 5.0g
KH
2
PO
4
1.0g
K
2
HPO
4
1.0g
MgSO
4
·7H
2
O 0.2g
NaCO
3
solution 100.0mL
pH 10.0 ± 0.2 at 25°C
NaCO
3
Solution:
Composition per 100.0mL:
NaCO

3
10.0g
Preparation of NaCO
3
Solution: Add NaCO
3
to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Filter ster-
ilize.
© 2010 by Taylor and Francis Group, LLC
854 Horse Blood Agar
Preparation of Medium: Add components, except NaCO
3
solu-
tion, to double distilled/deionized water and bring volume to 900.0mL.
Mix thoroughly. Adjust pH to 10.0. Gently heat while stirring and bring
to boiling. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 50°C. Aseptically add 100.0mL NaCO
3
solution. Ad-
just pH to 10.0. Pour into Petri dishes or aseptically distribute into
tubes.
Use: For the cultivation of Salinicoccus alkaliphilus.
Horse Blood Agar
Composition per liter:
Beef heart, infusion from 500.0g
Agar 15.0g
Tryptose 10.0g
NaCl 5.0g
Horse blood, defibrinated 50.0mL

pH 6.8 ± 0.2 at 25°C
Preparation of Medium: Add components, except horse blood, to
distilled/deionized water and bring volume to 950.0mL. 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 add sterile horse
blood. Mix thoroughly. Pour into sterile Petri dishes or distribute into
sterile tubes.
Use: For the cultivation and maintenance of Yersinia pseudotubercu-
losis.
Horse Serum Agar
Composition per liter:
Agar 15.0g
Pancreatic digest of gelatin 5.0g
Beef extract 3.0g
Horse serum 200.0mL
pH 6.8 ± 0.2 at 25°C
Preparation of Medium: Add components, except horse serum, to
distilled/deionized water and bring volume to 800.0mL. 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 add sterile horse se-
rum. Mix thoroughly. Pour into sterile Petri dishes or distribute into
sterile tubes.
Use: For the cultivation and maintenance of Pseudomonas aeruginosa
and Streptobacillus moniliformis.
Horse Serum Broth
Composition per liter:
Pancreatic digest of gelatin 5.0g
Beef extract 3.0g
Horse serum 200.0mL
pH 6.8 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from BD Di-
agnostic Systems.
Preparation of Medium: Add components, except horse serum, to
distilled/deionized water and bring volume to 800.0mL. 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 add sterile horse se-
rum. Mix thoroughly. Aseptically distribute into sterile tubes or flasks.
Use: For the cultivation and maintenance of Pseudomonas aeruginosa
and Streptobacillus moniliformis.
Hottinger Broth
Composition per liter:
Fish peptone 20.0g
Yeast extract 2.0g
Tryptophan 1.0g
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 cultivation of less fastidious microorganisms and determina-
tion of indole as per USSR State Pharmacopoeia.
Howardella Medium
(DSMZ Medium 1085)
Composition per liter:
Casitone 20.0g
Yeast extract 5.0g
Na
2

HPO
4
5.0g
MgCl
2
·6H
2
O 1.1g
Urea 1.0g
Na-thioglycolate 0.75g
Resazurin 0.5mg
Urea solution 10.0mL
pH 7.4 ± 0.2 at 25°C
Urea Solution:
Composition per 10.0mL:
Urea 1.0g
Preparation of Urea Solution: Add urea to distilled/deionized wa-
ter and bring volume to 10.0mL. Mix thoroughly. Sparge with 100%
N
2
. Filter sterilize.
Preparation of Medium: Add components, except thioglycolate
and urea solution, to double distilled/deionized water and bring volume
to 990.0mL. Cool to room temperature while sparging with 80% N
2
+
20% CO
2
. Add thioglycolate. Mix thoroughly. Distribute into tubes or
bottles under an atmosphere of 80% N

2
+ 20% CO
2
. Autoclave for 15
min at 15 psi pressure–121°C. Cool to room temperature. Aseptically
add 10.0mL urea solution. Adjust pH to 7.4.
Use: For the cultivation of Howardella spp.
Hoyer’s Medium
Composition per liter:
(NH
4
)
2
SO
4
1.0g
KH
2
PO
4
0.9g
MgSO
4
·7H
2
O 0.25g
K
2
HPO
4

0.1g
FeCl
3
·6H
2
O 0.02g
Ethanol solution 200.0mL
Ethanol Solution:
Composition
per 200.0mL:
Ethanol 30.0mL
Preparation of Ethanol Solution: Add ethanol to distilled/deion-
ized water and bring volume to 200.0mL. Mix thoroughly. Filter ster-
ilize.
© 2010 by Taylor and Francis Group, LLC