Potato Dextrose Yeast Agar

 

(PDY Agar)

Composition per liter:

Glucose ........................................................... .20.0g

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

Yeast extract..................................................... ..5.0g

Potato infusion.......................................... ..500.0mL

pH 5.6 ± 0.2 at 25oC

Potato Infusion:

Composition per 500.0mL:

Potatoes........................................................ ..300.0g

Preparation of Potato Infusion: Peel and dice
potatoes. Add 500.0mL of distilled/deionized water.
Gently heat and bring to boiling. Continue boiling for
30 min. Filter through cheesecloth. Reserve filtrate.

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-121oC. Pour into sterile Petri dish-
es or leave in tubes.

Use: For the cultivation and maintenance of Bacillus species and fungi.


Potato Extract Agar

Composition per liter:

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

Peptone ........................................................... ...5.0g

NaCl.................................................................. ..5.0g

Yeast extract...................................................... ..2.0g

Beef extract powder.......................................... ..1.0g

Potato extract ............................................. ... 20.0mL

pH 7.4 ± 0.2 at 25oC

Potato Extract:

Composition per liter:

Potatoes......................................................... ..300.0g

Preparation of Potato Extract: Peel and dice potatoes. Add 500.0mL of distilled/deionized water. Gently heat and bring to boiling. Continue boiling for 30 min. Filter through cheesecloth.

Use: For the cultivation of a wide variety of yeasts and molds.


PPLO Agar

Composition per liter:

Beef heart, infusion from................................. .50.0g

Agar .............................................................. ...14.0g

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

NaCl.................................................................. ..5.0g

Bovine serum............................................. . 100.0mL

pH 7.8 ± 0.2 at 25oC

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

Preparation of Medium: Add components, ex-
cept bovine serum, 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-121oC. Cool to 45o-50oC. Aseptically
add sterile bovine serum. Mix thoroughly. Pour into
sterile Petri dishes or distribute into sterile tubes.

Use: For the isolation and cultivation of Mycoplas-
ma species (pleuro-pneumonia-like organisms).


PPLO Agar

Composition per liter:

Beef heart, infusion from................................. .50.0g

Agar .............................................................. ...14.0g

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

NaCl................................................................. ..5.0g

Ascitic fluid............................................... .250.0mL

pH 7.8 ± 0.2 at 25oC

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

Preparation of Medium: Add components, ex-
cept ascitic fluid, to distilled/deionized water and
bring volume to 750.0mL. Mix thoroughly. Gently
heat and bring to boiling. Autoclave for 15 min at 15
psi pressure-121oC. Cool to 45o-50oC. Aseptically
add sterile ascitic fluid. Mix thoroughly. Pour into
sterile Petri dishes or distribute into sterile tubes.

Use: For the isolation and cultivation of Mycoplas-
ma species (pleuro-pneumonia-like organisms).


PPLO Broth with Crystal Violet

Composition per liter:

Beef heart, infusion from................................. .50.0g

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

NaCl................................................................. ..5.0g

Crystal Violet................................................... .0.01g

Ascitic fluid............................................... .250.0mL

Chapman tellurite solution........................... .2.85mL

pH 7.8 ± 0.2 at 25oC

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

Chapman Tellurite Solution:

Composition per 100.0mL:

K2TeO3.............................................................. .1.0g

Preparation of Chapman Tellurite Solution:
Add K2TeO3 to distilled/deionized water and bring
volume to 100.0mL. Mix thoroughly. Filter sterilize.

Caution: Potassium tellurite is toxic.

Preparation of Medium: Add components, ex-
cept ascitic fluid and Chapman tellurite solution, to distilled/deionized  water  and  bring  volume  to 747.15mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure-121oC. Cool to less than 37oC. Asep-
tically add sterile ascitic fluid and 2.85mL of Chap-
man tellurite solution. Mix thoroughly. Aseptically distribute into sterile tubes or flasks.

Use: For the isolation of Mycoplasma species from clinical specimens.


PPLO Broth without Crystal Violet

Composition per liter:

Beef heart, infusion from................................. .50.0g

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

NaCl................................................................. ..5.0g

Thallium acetate (optional)............................... ...0.5g 

Penicillin (optional).................................. ..100,000U

Ascitic fluid ............................................. ... 250.0mL

pH 7.8 ± 0.2 at 25oC

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

Preparation of Medium: Add components, ex-
cept ascitic fluid, to distilled/deionized water and
bring volume to 750.0mL. Mix thoroughly. Auto-
clave for 15 min at 15 psi pressure-121oC. Cool to
less than 37oC. Aseptically add sterile ascitic fluid. If
desired, 0.5g of thallium acetate or 100,000U of pen-
icillin may be added for a more selective medium.
Mix thoroughly. Aseptically distribute into sterile
tubes or flasks.

Use: For the enrichment of pleuro-pneumonia-like organisms (PPLOs) and Mycoplasma species from clinical specimens.


PPLO Broth without Crystal Violet

with Calf Serum, Fresh Yeast

Extract, and Sodium Acetate

Composition per liter:

Beef heart, infusion from................................. .50.0g

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

Sodium acetate................................................... .9.0g

NaCl.................................................................. ..5.0g

Yeast extract solution, fresh..................... ... 250.0mL

Calf serum................................................ ... 100.0mL

pH 7.8 ± 0.2 at 25oC

Yeast Extract Solution:

Composition per 300.0mL:

Baker’s yeast, live, pressed, starch-free......... ...75.0g

Preparation of Yeast Extract Solution: Add the live Baker’s yeast to 300.0mL of distilled/deion-
ized water. Autoclave for 90 min at 15 psi pressure-
121oC. Allow to stand. Remove supernatant solution. Adjust pH to 6.6-6.8. Filter sterilize.

Preparation of Medium: Add components, ex-
cept fresh yeast extract solution and calf serum, to
distilled/deionized  water  and  bring  volume  to
550.0mL. Mix thoroughly. Autoclave for 15 min at
15 psi pressure-121oC. Cool to 45o-50oC. Aseptical-
ly add sterile fresh yeast extract solution and calf se-
rum. Mix thoroughly. Aseptically distribute into
sterile tubes or flasks.

Use: For the cultivation and maintenance of Myco-
plasma species.


Presumpto Media

Composition per plate:

Quadrant 1 .................................................. ... 5.0mL

Quadrant 2 .................................................. ... 5.0mL

Quadrant 3..................................................... .5.0mL

Quadrant 4..................................................... .5.0mL

Quadrant 1:

Composition per 5.0mL:

Lombard-Dowell agar.................................... .5.0mL

Quadrant 2:

Composition per 5.0mL:

Lombard-Dowell bile agar........................... ...5.0mL

Quadrant 3:

Composition per 5.0mL:

Lombard-Dowell egg yolk agar..................... .5.0mL

Quadrant 4:

Composition per 5.0mL:

Lombard-Dowell esculin agar ..................... ...5.0mL

Preparation  of  Quadrant  Media:  Sterilize
Lombard-Dowell Agar by autocalving for 15 min at
15 psi pressure-121oC. Cool to 45o-50oC. Add addi-
tional components as filter sterilized solutions. Mix
and distribute as 5.0mL aliquots into quadrants.

Use: For the differentiation and presumptive identifi-
cation of anaerobic bacteria. The Presumpto media is a
four-sectored plate each containing a different medium.


Proteose No. 3 Agar

Composition per 1010.0mL:

Proteose peptone no. 3.................................... ..20.0g

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

Na2HPO4........................................................... .5.0g

NaCl................................................................. ..5.0g

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

Hemoglobin solution................................ ..500.0mL

Supplement A ............................................ ..10.0mL

pH 7.3 ± 0.2 at 25oC

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

Hemoglobin Solution:

Composition per 500.0mL:

Hemoglobin .................................................. ...10.0g

Preparation of Hemoglobin Solution: Add he-
moglobin to distilled/deionized water and bring vol-
ume to 500.0mL. Mix thoroughly. Autoclave for 15
min at 15 psi pressure-121oC. Cool to 45o-50oC.

Supplement A:

Composition per 100.mL:

Supplement A contains yeast concentrate with Crys-
      tal Violet.

Preparation of Supplement A: Add compo-
nents to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.


 

Preparation of Medium: Add components, ex-
cept hemoglobin solution and supplement A, to dis-
tilled/deionized water and bring volume to 500.0mL.
Mix thoroughly. Gently heat and bring to boiling.
Autoclave for 15 min at 15 psi pressure-121oC. Cool
to 50o-60oC. Aseptically add 500.0mL of sterile he-
moglobin solution and 10.0mL of sterile supplement
A. Mix thoroughly. Pour into sterile Petri dishes or
distribute into sterile tubes.

Use: For the isolation and cultivation of Neisseria species, Hemophilus species, and other fastidious bacteria. For the cultivation and maintenance of Es-
cherichia coli.


Proteose No. 3 Agar

Composition per 1010.0mL:

Proteose peptone no. 3.................................... ..20.0g

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

Na2HPO4........................................................... .5.0g

NaCl.................................................................. ..5.0g

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

Hemoglobin solution ................................. . 500.0mL

Supplement B............................................. ... 10.0mL

pH 7.3 ± 0.2 at 25oC

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

Hemoglobin Solution:

Composition per 500.0mL:

Hemoglobin .................................................. ...10.0g

Preparation of Hemoglobin Solution: Add he-
moglobin to distilled/deionized water and bring vol-
ume to 500.0mL. Mix thoroughly. Autoclave for 15
min at 15 psi pressure-121oC. Cool to 45o-50oC.

Supplement B:

Composition per 10.0mL:

Supplement B contains yeast concentrate, glutamine,
       coenzyme, cocarboxylase, hematin, and growth
       factors.

Preparation of Supplement B: Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, ex-
cept hemoglobin solution and supplement B, to dis-
tilled/deionized water and bring volume to 500.0mL.
Mix thoroughly. Gently heat and bring to boiling.
Autoclave for 15 min at 15 psi pressure-121oC. Cool
to 50o-60oC. Aseptically add 500.0mL of sterile he-
moglobin solution and 10.0mL of sterile supplement
B. Mix thoroughly. Pour into sterile Petri dishes or
distribute into sterile tubes.

Use: For the isolation and cultivation of Neisseria

species, Hemophilus species, and other fastidious bacteria. For the cultivation and maintenance of Es-
cherichia coli.


Proteose No. 3 Agar

Composition per 1010.0mL:

Proteose peptone no. 3.................................... ..20.0g

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

Na2HPO4........................................................... .5.0g

NaCl................................................................. ..5.0g

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

Hemoglobin solution................................ ..500.0mL

Supplement VX......................................... ...10.0mL

pH 7.3 ± 0.2 at 25oC

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

Hemoglobin Solution:

Composition per 500.0mL:

Hemoglobin .................................................. ...10.0g

Preparation of Hemoglobin Solution: Add he-
moglobin to distilled/deionized water and bring vol-
ume to 500.0mL. Mix thoroughly. Autoclave for 15
min at 15 psi pressure-121oC. Cool to 45o-50oC.

Supplement VX:

Composition per 10.0mL:

Supplement B contains essential growth factors.
Preparation of Supplement VX: Add compo-
nents to distilled/deionized water and bring volume
to 10.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium: Add components, ex-
cept hemoglobin solution and supplement VX, to dis-
tilled/deionized water and bring volume to 500.0mL.
Mix thoroughly. Gently heat and bring to boiling.
Autoclave for 15 min at 15 psi pressure-121oC. Cool
to 50o-60oC. Aseptically add 500.0mL of sterile he-
moglobin solution and 10.0mL of sterile supplement
VX. Mix thoroughly. Pour into sterile Petri dishes or
distribute into sterile tubes.

Use: For the isolation and cultivation of Neisseria species, Hemophilus species, and other fastidious bacteria. For the cultivation and maintenance of Es-
cherichia coli.


Proteose Yeast Extract Medium

 Composition per liter:

Proteose peptone............................................. ..20.0g

Glucose ........................................................... .10.0g

Yeast extract..................................................... ..5.0g

Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.

Mix thoroughly. Distribute into tubes or flasks. Auto-

clave for 15 min at 15 psi pressure-121oC.
Use: For the cultivation of a variety of bacteria.


Pseudomonas Agar F

Composition per liter:

Proteose peptone no. 3.................................... ..20.0g

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

Glycerol ........................................................ ...10.0g

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

K2HPO4........................................................... ...1.5g

MgSO4*7H2O.................................................. .0.73g

pH 7.0 ± 0.2 at 25oC

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-121oC. Pour into sterile Petri dish-
es or leave in tubes.

Use: For the cultivation and observation of fluores-
cein production in Pseudomonas species.


Pseudomonas Agar F

Composition per liter:

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

Glycerol ........................................................ ...10.0g

Proteose peptone no. 3.................................... ..10.0g

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

K2HPO4........................................................... ...1.5g

MgSO4*7H2O.................................................. ...1.5g

pH 7.0 ± 0.2 at 25oC