Middlebrook 7H10 Agar

with Streptomycin

Composition per liter:

Agar .............................................................. ...15.0g

Na2HPO4........................................................... .1.5g

KH2PO4........................................................... ...1.5g

(NH4)2SO4........................................................ ..0.5g

L-Glutamic acid................................................ ...0.5g

Sodium citrate................................................... ..0.4g

Ferric ammonium citrate................................. ..0.04g

MgSO4*7H2O............................................... ..0.025g

ZnSO4*7H2O ................................................. .1.0mg

CuSO4*5H2O.................................................. .1.0mg

Pyridoxine..................................................... ..1.0mg

Biotin ........................................................... ...0.5mg

CaCl2*2H2O.................................................. ...0.5mg

Malachite Green.......................................... ...0.25mg

Glycerol ........................................................ .5.0mL

Streptomycin............................................. ...100.0mg

pH 6.6 ± 0.2 at 25oC

Source: This medium is available as a premixed powder from  BD Diagnostic Systems.

Preparation of Medium: Add glycerol to 1.0L of
distilled/deionized water and add remaining compo-
nents. Mix thoroughly. Gently heat and bring to boil-
ing. Autoclave for 15 min at 15 psi pressure-121oC.
Cool to 50o-55oC. Aseptically add streptomycin.
Mix thoroughly. Pour into sterile Petri dishes or dis-
tribute into sterile tubes.

Use: For the isolation, cultivation, and maintenance of Mycobacterium kansasii.


Middlebrook 7H11 Agar, Selective

 Composition per liter:

Agar .............................................................. ...15.0g

Na2HPO4........................................................... .1.5g

KH2PO4........................................................... ...1.5g

Pancreatic digest of casein.................................. .1.0g

(NH4)2SO4 ........................................................ .0.5g

L-Glutamic acid ................................................ ..0.5g

Sodium citrate................................................... ..0.4g

MgSO4*7H2O.................................................. .0.05g

Ferric ammonium citrate.................................. ..0.04g

Pyridoxine...................................................... ..1.0mg

ZnSO4*7H2O.................................................. .1.0mg

CuSO4*5H2O.................................................. .1.0mg

CaCl2*2H2O .................................................. ..0.5mg

Malachite Green.......................................... ...0.25mg

D-Biotin ......................................................... ...0.5ตg

Middlebrook OADC enrichment............... . 100.0mL

Antibiotic solution ....................................... . 10.0mL

Glycerol ........................................................ . 5.0mL

pH 6.6 ± 0.2 at 25oC

Middlebrook OADC Enrichment: Composition per 100.0mL:

Bovine albumin fraction V .............................. ...5.0g

Glucose ........................................................... ...2.0g

NaCl............................................................... ...0.85g

Oleic acid......................................................... .0.05g

Catalase......................................................... ...4.0mg

Source: This enrichment is available as a prepared enrichment from  BD Diagnostic Systems.

Preparation of Middlebrook OADC Enrich-
ment: Add components to distilled/deionized water
and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize.

Antibiotic Solution:

Composition per 10.0mL:

Carbenicillin ................................................ ..0.05mg

Trimethoprim lactate..................................... ..0.02mg

Amphotericin B .......................................... ...0.01mg

Polymyxin B............................................. .200,000U

Preparation of Antibiotic Solution: Add com-

ponents to distilled/deionized water and bring vol-
ume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation  of  Medium:  Add  glycerol  to 890.0mL of distilled/deionized water and add re-
maining components, except Middlebrook OADC enrichment and antibiotic solution. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure-121oC. Cool to 50o-55oC. Aseptically add 100.0mL of sterile Middlebrook

OADC enrichment and 10.0mL of sterile antibiotic solution. Mix thoroughly. Pour into sterile Petri dish-
es or distribute into sterile tubes.

Use: For the selective isolation and cultivation of pathogenic mycobacteria from specimens potentially contaminated with bacteria and fungi.


Middlebrook 7H11 Agar with

Middlebrook ADC Enrichment

(Mycobacteria 7H11 Agar with
       Middlebrook ADC Enrichment)
Composition per liter:

Agar .............................................................. ...15.0g

Na2HPO4........................................................... .1.5g

KH2PO4........................................................... ...1.5g

Pancreatic digest of casein................................. .1.0g

(NH4)2SO4........................................................ ..0.5g

L-Glutamic acid................................................ ...0.5g

Sodium citrate................................................... ..0.4g

MgSO4*7H2O.................................................. .0.05g

Ferric ammonium citrate................................. ..0.04g

Pyridoxine..................................................... ..1.0mg

Malachite Green.......................................... ...0.25mg

D-Biotin........................................................ ... 0.5ตg

Middlebrook ADC enrichment.................. .100.0mL

Glycerol ........................................................ .5.0mL

pH 6.6 ± 0.2 at 25oC

Source: This medium is available as a premixed powder from  BD Diagnostic Systems.

Middlebrook ADC Enrichment:

Composition per 100.0mL:

Bovine albumin fraction V .............................. ...5.0g

Glucose ........................................................... ...2.0g

Catalase......................................................... ..0.003g

Source: This enrichment is available as a prepared enrichment from  BD Diagnostic Systems.

Preparation of Middlebrook ADC Enrich-
ment: Add components to distilled/deionized water
and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize.

Preparation  of  Medium:  Add  glycerol  to
900.0mL of distilled/deionized water and add re-

maining components, except Middlebrook ADC en-

richment. Mix thoroughly. Gently heat and bring to
boiling. Autoclave for 15 min at 15 psi pressure-
121oC. Cool to 50o-55oC. Aseptically add 100.0mL
of sterile Middlebrook ADC enrichment. Mix thor-
oughly. Pour into sterile Petri dishes or distribute into
sterile tubes.

Use: For the cultivation of drug-resistant (isoniazid
[INH]) strains of Mycobacterium tuberculosis. For
the cultivation of particularly fastidious strains of
tubercle bacilli that occur following treatment of
tuberculosis patients with secondary antitubercular
drugs. Generally, these strains fail to grow on 7H10
medium.


Middlebrook 7H11 Agar with
Middlebrook OADC Enrichment

(Mycobacteria 7H11 Agar with
     Middlebrook OADC Enrichment)
Composition per liter:

Agar .............................................................. ...15.0g

Na2HPO4........................................................... .1.5g

KH2PO4........................................................... ...1.5g

Pancreatic digest of casein.................................. .1.0g

(NH4)2SO4 ........................................................ .0.5g

L-Glutamic acid ................................................ ..0.5g

Sodium citrate................................................... ..0.4g

MgSO4*7H2O.................................................. .0.05g

Ferric ammonium citrate.................................. ..0.04g

Pyridoxine...................................................... ..1.0mg

Malachite Green.......................................... ...0.25mg

D-Biotin............................................................ .0.5ตg

Middlebrook OADC enrichment............... . 100.0mL

Glycerol ........................................................ . 5.0mL

pH 6.6 ± 0.2 at 25oC

Source: This medium is available as a premixed powder from  BD Diagnostic Systems.

Middlebrook OADC Enrichment: Composition per 100.0mL:

Bovine albumin fraction V .............................. ...5.0g

Glucose ........................................................... ...2.0g

NaCl............................................................... ...0.85g

Oleic acid......................................................... .0.05g

Catalase......................................................... ...4.0mg

Source: This enrichment is available as a prepared enrichment from  BD Diagnostic Systems.

Preparation of Middlebrook OADC Enrich-
ment: Add components to distilled/deionized water
and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize.

Preparation  of  Medium:  Add  glycerol  to
900.0mL of distilled/deionized water and add re-

maining components, except Middlebrook OADC

enrichment. Mix thoroughly. Gently heat and bring to
boiling. Autoclave for 15 min at 15 psi pressure-
121oC. Cool to 50o-55oC. Aseptically add 100.0mL
of sterile Middlebrook OADC enrichment. Mix thor-
oughly. Pour into sterile Petri dishes or distribute into
sterile tubes.

Use: For the cultivation of drug-resistant (isoniazid
[INH]) strains of Mycobacterium tuberculosis. For
the cultivation of particularly fastidious strains of
tubercle bacilli that occur following treatment of
tuberculosis patients with secondary antitubercular
drugs. Generally, these strains fail to grow on 7H10
medium.


Middlebrook 7H11 Agar with

Middlebrook OADC Enrichment

and Triton WR 1339

(Mycobacteria 7H11 Agar with
Middlebrook OADC Enrichment

and Triton WR 1339)

Composition per liter:

Agar .............................................................. ...15.0g

Na2HPO4........................................................... .1.5g

KH2PO4........................................................... ...1.5g

Pancreatic digest of casein................................. .1.0g

(NH4)2SO4........................................................ ..0.5g

L-Glutamic acid................................................ ...0.5g

Sodium citrate................................................... ..0.4g

MgSO4*7H2O.................................................. .0.05g

Ferric ammonium citrate................................. ..0.04g

Pyridoxine..................................................... ..1.0mg

Malachite Green.......................................... ...0.25mg

D-Biotin........................................................ ... 0.5ตg

Middlebrook OADC enrichment

with Triton WR 1339.......................... .100.0mL

Glycerol ........................................................ .5.0mL

pH 6.6 ± 0.2 at 25oC

Source: This medium is available as a premixed powder from  BD Diagnostic Systems.

Middlebrook OADC Enrichment with Triton WR 1339:

Composition per 100.0mL:

Bovine albumin fraction V .............................. ...5.0g

Glucose ........................................................... ...2.0g

NaCl.............................................................. ...0.85g

Triton WR 1339............................................. ...0.25g

Oleic acid......................................................... .0.05g

Catalase......................................................... ...4.0mg

Source: This enrichment is available as a prepared enrichment from  BD Diagnostic Systems.

Preparation of Middlebrook OADC Enrich-
ment with Triton WR 1339: Add components to distilled/deionized  water  and  bring  volume  to 100.0mL. Mix thoroughly. Filter sterilize.

Preparation  of  Medium:  Add  glycerol  to 900.0mL of distilled/deionized water and add re-
maining components, except Middlebrook OADC enrichment with Triton WR-1339. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure-121oC. Cool to 50o-55oC. Aseptically add 100.0mL of sterile Middlebrook

OADC enrichment with Triton WR-1339. Mix thor-
oughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation of drug-resistant (isoniazid [INH]) strains of Mycobacterium tuberculosis. For the cultivation of particularly fastidious strains of tubercle bacilli that occur following treatment of tuberculosis patients with secondary antitubercular drugs. Generally, these strains fail to grow on 7H10 medium.


Middlebrook 7H12 Medium

Composition per 102.5mL:

Bovine serum albumin....................................... .0.5g

Casein hydrolyslate.......................................... ...0.1g

Catalase......................................................... ..4800U

14C-Palmitic acid .......................................... .100ตCi

Middlebrook 7H9 broth........................... ... 100.0mL

Antibiotic solution ....................................... ... 2.5mL

pH 6.8 ± 0.1 at 25oC

Middlebrook 7H9 Broth:

Composition per liter:

Na2HPO4........................................................... .2.5g

KH2PO4........................................................... ...1.0g

Monosodium glutamate .................................... ..0.5g

(NH4)2SO4 ........................................................ .0.5g

Sodium citrate................................................... ..0.1g

MgSO4*7H2O.................................................. .0.05g

Ferric ammonium citrate.................................. ..0.04g

CuSO4*5H2O.................................................. .1.0mg

Pyridoxine...................................................... ..1.0mg

ZnSO4*7H2O.................................................. .1.0mg

Biotin ........................................................... ...0.5mg

CaCl2*2H2O .................................................. ..0.5mg

Glycerol ........................................................ . 2.0mL

Preparation of Middlebrook 7H9 Broth: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

Antibiotic Solution:

Composition per 5.0mL:

Nalidixic acid................................................... ...0.2g

Azlocillin ......................................................... ...0.1g

Amphotericin B ............................................. ...0.05g

Trimethoprim................................................... .0.05g

Polymyxin B ............................................ .500,000U

Preparation of Antibiotic Solution: Add com-
ponents to distilled/deionized water and bring vol-
ume to 5.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: To 100.0mL of Middle-
brook 7H9 broth, add remaining components, except antibiotic solution. Mix thoroughly. Filter sterilize. Aseptically distribute into bottles in 4.0mL volumes. Prior to inoculation, aseptically add 0.1mL of antibi-
otic solution to each bottle. Mix thoroughly.

Use: For the cultivation of Mycobacterium species from the blood of patients suspected of having myco-
bacteremia.


Middlebrook Medium

(DSMZ Medium 645)

Composition per liter:

Agar .............................................................. ...15.0g

Na2HPO4........................................................... .1.5g

KH2PO4........................................................... ...1.5g

(NH4)2SO4........................................................ ..0.5g

L-Glutamic acid................................................ ...0.5g

Sodium citrate................................................... ..0.4g

Ferric ammonium citrate................................. ..0.04g

MgSO4*7H2O............................................... ..0.025g

ZnSO4*7H2O ................................................. .1.0mg

CuSO4*5H2O.................................................. .1.0mg

Pyridoxine..................................................... ..1.0mg

Biotin ........................................................... ...0.5mg

CaCl2*2H2O.................................................. ...0.5mg

Malachite Green.......................................... ...0.25mg

Middlebrook OADC enrichment .............. .100.0mL

Glycerol ........................................................ .5.0mL

pH 6.6 ± 0.2 at 25oC

Source: This medium is available as a premixed powder from  BD Diagnostic Systems.

Middlebrook OADC Enrichment:

Composition per 100.0mL:

Bovine albumin fraction V .............................. ...5.0g

Glucose ........................................................... ...2.0g

Catalase......................................................... ..0.003g

Distilled water.......................................... ...100.0mL

Source: This enrichment is available as a prepared enrichment from  BD Diagnostic Systems.

Preparation of Middlebrook OADC Enrich-
ment: Add components to distilled/deionized water
and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize.

Preparation  of  Medium:  Add  glycerol  to
900.0mL of distilled/deionized water and add re-
maining components, except Middlebrook OADC
 

enrichment. Mix thoroughly. Gently heat and bring to

boiling. Autoclave for 15 min at 15 psi pressure-
121oC. Cool to 50o-55oC. Aseptically add 100.0mL
of sterile Middlebrook OADC enrichment. Mix thor-
oughly. Pour into sterile Petri dishes or distribute into
sterile tubes.

Use: For the isolation, cultivation, and maintenance of Mycobacterium species.


Middlebrook Medium with Mycobactin

(DSMZ Medium 780)

Composition per liter:

Agar .............................................................. ...15.0g

Na2HPO4........................................................... .1.5g

KH2PO4........................................................... ...1.5g

(NH4)2SO4 ........................................................ .0.5g

L-Glutamic acid................................................ ...0.5g

Sodium citrate................................................... ..0.4g

Ferric ammonium citrate.................................. ..0.04g

MgSO4*7H2O............................................... ..0.025g

Mycobactin J................................................... .2.0mg

ZnSO4*7H2O.................................................. .1.0mg

CuSO4*5H2O.................................................. .1.0mg

Pyridoxine...................................................... ..1.0mg

Biotin ........................................................... ...0.5mg

CaCl2*2H2O .................................................. ..0.5mg

Malachite Green.......................................... ...0.25mg

Middlebrook ADC enrichment.................. . 100.0mL

Glycerol ........................................................ . 5.0mL

pH 6.6 ± 0.2 at 25oC

Source: Mycobactin J is available from Allied Lab-
oratories, Inc.

Middlebrook ADC Enrichment: Composition per 100.0mL:

Bovine albumin fraction V .............................. ...5.0g

Glucose ........................................................... ...2.0g

Catalase......................................................... ..0.003g

Distilled water.......................................... ... 100.0mL

Source: This medium and enrichment is available from  BD Diagnostic Systems.

Preparation of Middlebrook ADC Enrich-
ment: Add components to distilled/deionized water
and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize.

Preparation  of  Medium:  Add  glycerol  to
900.0mL of distilled/deionized water and add re-
maining components, except Middlebrook ADC en-
richment and mycobactin. Mix thoroughly. Gently
heat and bring to boiling. Autoclave for 15 min at 15
psi pressure-121oC. Cool to 50o-55oC. Aseptically
add 100.0mL of sterile Middlebrook ADC enrich-
ment and mycobactin. The mycobactin is dissolved

in 2.0mL ethanol. Be sure to add all of the mycobac- 

MIL Medium

tin; wash with additional 2.0mL ethanol if needed.

(Motility Indole Lysine Medium)

Mix thoroughly. Pour into sterile Petri dishes or dis-

tribute into sterile tubes.

Use: For the cultivation of Mycobacterium avium subsp. paratuberculosis.


Middlebrook OADC Enrichment

(Middlebrook Oleic Albumin

Dextrose Catalase Enrichment) Composition per 100.0mL:

Bovine albumin fraction V .............................. ...5.0g

Glucose ........................................................... ...2.0g

NaCl.............................................................. ...0.85g

Oleic acid......................................................... .0.05g

Catalase......................................................... ...4.0mg

Source: This medium is available as a prepared en-
richment from  BD Diagnostic Systems.

Preparation of Enrichment: Add components to distilled/deionized  water  and  bring  volume  to 100.0mL. Mix thoroughly. Filter sterilize.

Use: For use as a supplement to other Middlebrook
media for the isolation, cultivation, and maintenance
of Mycobacterium species. Also used as a supple-
ment to other Middlebrook media for determining
the antimicrobial susceptibility of mycobacteria.


Middlebrook OADC Enrichment

with Triton WR 1339

(Middlebrook Oleic Albumin

Dextrose Catalase Enrichment

with Triton WR 1339)

Composition per 100.0mL:

Bovine albumin fraction V .............................. ...5.0g

Glucose ........................................................... ...2.0g

NaCl.............................................................. ...0.85g

Triton WR 1339............................................. ...0.25g

Oleic acid......................................................... .0.05g

Catalase......................................................... ...4.0mg

Source: This medium is available as a prepared en-
richment from  BD Diagnostic Systems.

Preparation of Enrichment: Add components to distilled/deionized  water  and  bring  volume  to 100.0mL. Mix thoroughly. Filter sterilize.

Use: For use as a supplement to other Middlebrook
media for the isolation, cultivation, and maintenance
of Mycobacterium species. Also used as a supple-
ment to other Middlebrook media for determining
the antimicrobial susceptibility of mycobacteria.

Composition per liter:

Peptone ........................................................... .10.0g

Pancreatic digest of casein............................... ..10.0g

L-Lysine*HCl.................................................. ..10.0g

Yeast extract...................................................... ..3.0g

Agar ................................................................. ..2.0g

Dextrose............................................................ ..1.0g

Ferric ammonium citrate..................................... .0.5g

Bromcresol Purple ......................................... ..0.02g

pH 6.6 ± 0.2 at 25oC

Source: This medium is available as a premixed pow-
der and prepared medium from  BD Diagnostic Systems.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes in 5.0mL volumes. Autoclave for 15 min at 15 psi pressure-121oC.

Use: For the cultivation and differentiation of mem-
bers of the Enterobacteriaceae on the basis of motil-
ity, lysine decarboxylase activity, lysine deaminase activity, and indole production.


Minimum Essential Medium with
   Bicarbonate, Serum, and Antibiotics
Composition per 1100.0mL:

Minimum essential medium ..................... .. 950.0mL

Fetal bovine serum, heat inactivated .......... . 100.0mL

NaHCO3 solution....................................... ... 40.0mL

Penicillin-streptomycin solution................... . 10.0mL

pH 7.2 ± 0.2 at 25oC

Minimum Essential Medium (MEM): Composition per liter:

Inorganic salt solution.............................. ... 400.0mL

Other component solution........................ ... 400.0mL

Amino acid solution................................. ... 100.0mL

Vitamin solution....................................... ... 100.0mL

Inorganic Salt Solution:

Composition per 400.0mL:

NaCl.................................................................. ..6.8g

KCl..................................................................... .0.4g

CaCl2, anhydrous............................................. ...0.2g

NaH2PO4*H2O............................................... ...0.14g

MgSO4, anhydrous................................... ...97.67mg

Preparation of Inorganic Salt Solution: Add
components to distilled/deionized water and bring
volume to 400.0mL. Mix thoroughly. Autoclave for
15 min at 15 psi pressure-121oC. Cool to 25oC.

Other Component Solution:

Composition per 400.0mL:

D-Glucose........................................................ ...1.0g

Phenol Red.................................................. ..10.0mg

Preparation of Other Component Solution:

Add components to distilled/deionized water and
bring volume to 400.0mL. Mix thoroughly. Auto-
clave for 15 min at 15 psi pressure-121oC. Cool to
25oC.

Amino Acid Solution:

Composition per 100.0mL:

L-Glutamine ............................................... .292.0mg

L-Arginine*HCl ........................................ ..126.1mg

L-Lysine*HCl............................................... ..72.5mg

L-Isoleucine.................................................. .52.0mg

L-Leucine ..................................................... .52.0mg

L-Tyrosine, disodium salt............................. .52.0mg

L-Threonine.................................................. .48.0mg

L-Valine........................................................ .46.0mg

L-Histidine*HCl*H2O.................................. ..42.0mg

L-Phenylalanine ......................................... ...32.0mg

L-Cysteine*2HCl ......................................... ..31.0mg

L-Methionine............................................... ..15.0mg

L-Glutamic acid............................................. .14.7mg

L-Aspartic acid............................................. ..13.3mg

L-Asparagine*H2O...................................... ...13.2mg

L-Proline..................................................... ...11.5mg

L-Serine........................................................ .10.5mg

L-Tryptophan............................................... ..10.0mg

L-Alanine ..................................................... ...8.9mg

Glycine........................................................... .7.5mg

Preparation of Amino Acid Solution: Add
components to distilled/deionized water and bring
volume to 100.0mL. Mix thoroughly. Filter sterilize.

Vitamin Solution:

Composition per 100.0mL:

i-Inositol........................................................ ..2.0mg

D-Ca pantothenate.......................................... ..1.0mg

Choline chloride............................................. ..1.0mg

Folic acid........................................................ .1.0mg

Niacinamide................................................... ..1.0mg

Pyridoxal*HCl .............................................. ..1.0mg

Thiamine*HCl ............................................... ..1.0mg

Riboflavin ..................................................... ..0.1mg

Preparation of Vitamin Solution: Add compo-
nents to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

Preparation of Minimum Essential Medium (MEM): Aseptically combine 400.0mL of sterile inorganic salt solution, 400.0mL of sterile other com-
ponent solution, 100.0mL of sterile amino acid solu-
tion, and 100.0mL of sterile vitamin solution. 

NaHCO3 Solution:

Composition per 40.0mL:

NaHCO3........................................................... ..2.0g

Preparation  of  NaHCO3  Solution:  Add NaHCO3 to distilled/deionized water and bring vol-
ume to 40.0mL. Mix thoroughly. Filter sterilize.

Penicillin-Streptomycin Solution: Composition per 10.0mL:

Penicillin......................................................... ..0.01g

Streptomycin................................................... ..0.01g

Preparation of Penicillin-Streptomycin Solu-
tion: Add components to distilled/deionized water
and bring volume to 10.0mL. Mix thoroughly. Filter
sterilize.

Preparation of Medium: Aseptically combine
950.0mL of  sterile minimum essential  medium,
100.0mL of sterile heat inactivated fetal bovine se-
rum, 40.0mL  of  sterile  NaHCO3  solution,  and

10.0mL of sterile penicillin-streptomycin solution.
Adjust pH to 7.2 with humidified 10% CO2 in 90%
air.

Use: For the cultivation of Encephalitozoon cuniculi, Encephalitozoon hellem, Encephalitozoon intestina-
lis, Naegleria fowleri, and Nosema corneum.


Mitis-Salivarius Agar

Composition per liter:

Sucrose........................................................... ..50.0g

Agar .............................................................. ...15.0g

Enzymatic digest of protein............................ ...10.0g

Proteose peptone............................................. ..10.0g

K2HPO4........................................................... ...4.0g

Dextrose............................................................ ..1.0g

Trypan Blue .................................................. ...0.08g

Crystal Violet................................................ ...0.8mg

Na2TeO3 solution.......................................... .. 1.0mL

pH 7.0 ± 0.2 at 25oC

Source: This medium is available as a premixed powder from  BD Diagnostic Systems.
Na2TeO3 Solution:

Composition per 10.0mL:

Na2TeO3............................................................ ..0.1g

Preparation  of  Na2TeO3  Solution:  Add Na2TeO3 to 10.0mL of distilled/deionized water. Mix thoroughly. Filter sterilize.

Caution: Potassium tellurite is toxic.

Preparation of Medium: Add components to
distilled/deionized  water  and  bring  volume  to
999.0mL. Mix thoroughly. Gently heat and bring to
boiling. Autoclave for 15 min at 15 psi pressure-
121oC. Cool medium to 50o-55oC. Aseptically add

1.0mL of the sterile Na2TeO3 solution to the cooled

basal medium. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the selective isolation of Streptococcus mitis, Streptococcus salivarius, and other viridans streptococci and enterococci.

M๘ller Decarboxylase Broth

Composition per liter:

Amino acid..................................................... ..10.0g

Peptic digest of animal tissue........................... ...5.0g

Beef extract...................................................... ...5.0g

Glucose ........................................................... ...0.5g

Bromcresol Purple ......................................... ..0.01g

Cresol Red..................................................... ..5.0mg

Pyridoxal........................................................ .5.0mg

pH 6.0 ± 0.2 at 25oC

Source: This medium is available as a premixed powder from  BD Diagnostic Systems.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Use L-lysine, L-arginine, or L-ornithine. Mix thor-
oughly. Gently heat until dissolved. Distribute into screw-capped tubes in 5.0mL volumes. Autoclave for 15 min at 15 psi pressure-121oC. A slight precip-
itate may form in the ornithine broth.

Use: For the differentiation of Gram-negative enteric
bacteria based on the production of arginine dihydro-
lase, lysine decarboxylase, or ornithine decarboxylase.


Molybdate Agar

Composition per101.5mL:

Base........................................................... .100.0mL

Phosphomolybdic acid solution..................... .1.5mL

pH 5.3 ± 0.2 at 25oC

Base:

Composition per liter:

Sucrose........................................................... ..40.0g

Agar .............................................................. ...15.0g

Meat peptone.................................................. ..10.0g

Preparation of Base: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.6. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure-
121oC. Cool to 45o-50oC.

Phosphomolybdic Acid Solution:

Composition per 100.0mL:

P2O5*2OMoO3.............................................. ...12.5g

Preparation of Phosphomolybdic Acid Solu-
tion: Add P2O5*2OMoO3 (phospho-12-molybdic ac-
id, 12-molybdophosphoric acid, or PMA) to sterile

distilled/deionized water. Mix thoroughly. Do not ad-
just pH.

Preparation of Medium: To 100.0mL of cooled sterile base, add 1.5mL of phosphomolybdic acid so-
lution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the isolation and presumptive identification
of yeast, especially Candida species. Candida albi-
cans appears as smooth, medium olive colonies with
medium olive bottoms. Candida stellatoidea appears
as shiny, light gray colonies with light gray bottoms.
Candida tropicalis appears as smooth, shiny, dark
blue/gray colonies with dark blue/gray bottoms.
Candida krusei appears as smooth, dull white colo-
nies with white bottoms. Saccharomyces cerevisiae
appears as smooth, shiny light blue/dark blue colo-
nies with dark blue/green bottoms.


Monsur Agar

(Taurocholate Tellurite Gelatin Agar) Composition per liter:

Gelatin............................................................ ...30.0g

Agar .............................................................. ...15.0g

Casein peptone................................................ ..10.0g

NaCl............................................................... ...10.0g

Sodium taurocholate........................................... .5.0g

Na2CO3 *H2O .................................................. ...1.0g

K2TeO3 solution.......................................... .. 10.0mL

pH 8.5 ± 0.2 at 25oC

K2TeO3 Solution:

Composition per 10.0mL:

K2TeO3............................................................ ..0.02g

Preparation of K2TeO3 Solution: Add K2TeO3 to 10.0mL of distilled/deionized water. Mix thor-
oughly. Filter sterilize.

Caution: Potassium tellurite is toxic.

Preparation of Medium: Add components, ex-
cept K2TeO3 solution, 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-121oC. Cool to 45o-50oC. Add 10.0mL
of sterile K2TeO3 solution. Mix thoroughly. Pour into
sterile Petri dishes or distribute into sterile tubes.
Use: For the isolation of Vibrio cholerae from fecal
specimens.