Oxidative-Fermentative

Glucose Medium, Semisolid

(OF Glucose Medium, Semisolid)

Composition per liter:

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

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

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

Pancreatic digest of casein................................. .2.0g

K2HPO4........................................................... ...0.3g

Bromthymol Blue dye.................................... ...0.08g

pH 6.8 ± 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  differentiating  Gram-negative  bacteria based upon determining the oxidative and fermenta-
tive metabolism of glucose. Bacteria that ferment glucose turn the medium yellow.


Oxidative-Fermentative

Glucose Medium, Semisolid, with NaCl
      (OF Glucose Medium, Semisolid with NaCl)

Composition per liter:

NaCl.............................................................. ...20.0g

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

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

Pancreatic digest of casein................................. .2.0g

K2HPO4........................................................... ...0.3g

Bromthymol Blue dye.................................... ...0.08g

pH 6.8 ± 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 differentiating Vibrio species based upon determining the oxidative and fermentative metabo-
lism of glucose. Bacteria that ferment glucose turn the medium yellow.


Oxidative-Fermentative Test Medium

(OF Test Medium)

Composition per liter:

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

Agar ................................................................. ..3.0g

Peptone.............................................................. .2.0g

K2HPO4........................................................... ...0.3g

Bromthymol Blue ... ............................................ ...0.03g

                            Carbohydrate solution.............................. ... 100.0mL

            Carbohydrate Solution:

                     Composition per 100.0mL:

Carbohydrate.................................................... ..10.0g

Preparation of Carbohydrate Solution: Add

carbohydrate to distilled/deionized water and bring
volume to 100.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium: Add components, ex-
cept carbohydrate solution, to distilled/deionized wa-
ter and bring volume to 1.0L. Mix thoroughly. Gently
heat and bring to boiling. Distribute into tubes in
3.0mL volumes. Autoclave for 15 min at 15 psi pres-
sure-121oC. Cool to 45o-50oC. Aseptically add

0.3mL of sterile carbohydrate solution to each tube. Mix thoroughly.

Use: For the cultivation and differentiation of a vari-
ety of microorganisms based on their ability to fer-
ment a specific carbohydrate. Bacteria that ferment
the specific carbohydrate turn the medium yellow.


P Agar

Composition per liter:

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

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

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

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

Glucose ........................................................... ...1.0g

pH 7.5 ± 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 of Staphylococcus species.

Pagano Levin Agar

Composition per liter:

Glucose ........................................................... .40.0g

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

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

Yeast extract...................................................... ..1.0g

Neomycin......................................................... ...0.5g

2,3,5-Triphenyltetrazolium chloride................... .0.1g

pH 6.0 ± 0.1 at 25oC

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

Preparation of Medium: Add components, ex-
cept neomycin and 2,3,5-triphenyltetrazolium chlo-
ride, to distilled/deionized water and bring volume to

1.0L. Mix thoroughly. Gently heat and bring to boil-

ing. Autoclave for 15 min at 15 psi pressure-121oC. Cool to 45o-50oC. Aseptically add neomycin and 2,3,5-triphenyltetrazolium chloride. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Allow tubes to cool in a slanted position.
Use: For the isolation, cultivation, and differentiation of Candida species. Candida albicans appears as smooth, shiny, cream-light pink colonies.


Pai Medium

Composition per liter:

Homogenized whole egg .......................... .666.0mL

NaCl (0.85% solution).............................. ..334.0mL

pH 6.75 ± 0.2 at 25oC

Homogenized Whole Egg:

Composition per liter:

Whole eggs .................................................. ... 18-24

Preparation of Homogenized Whole Egg: Use
fresh eggs, less than 1 week old. Scrub the shells with
soap. Let stand in a soap solution for 30 min. Rinse in
running water. Soak eggs in 70% ethanol for 15 min.
Break the eggs into a sterile container. Homogenize by
shaking. Filter through four layers of sterile cheesecloth
into a sterile graduated cylinder. Measure out 1.0L.
Preparation of Medium: Combine components.
Mix thoroughly. Aseptically distribute into sterile
tubes. Inspissate tubes in a slanted position at 80o-
90oC (moist heat) for 30 min.

Use: For the maintenance of stock cultures of Sal-
monella typhi and other Salmonella species.


Pai Medium

Composition per 1620.0mL:

Glucose ........................................................... ...5.0g

Homogenized whole egg ................................ .. 1.0L

Glycerol .................................................. ...120.0mL

pH 6.75 ± 0.2 at 25oC

Homogenized Whole Egg:

Composition per liter:

Whole eggs .................................................. ...18-24

Preparation of Homogenized Whole Egg:

Use fresh eggs, less than 1 week old. Scrub the shells
with soap. Let stand in a soap solution for 30 min.
Rinse in running water. Soak eggs in 70% ethanol for
15 min. Break the eggs into a sterile container. Ho-
mogenize by shaking. Filter through four layers of
sterile cheesecloth into a sterile graduated cylinder.
Measure out 1.0L.

Preparation of Medium: Combine components.
Mix thoroughly. Aseptically distribute into sterile

tubes. Inspissate tubes in a slanted position at 80o-
90oC (moist heat) for 30 min.

Use: For the isolation and cultivation of Corynebac-
terium spp.


PALCAM Agar

(Polymyxin Acriflavine LiCl
   Ceftazidime Esculin Mannitol Agar)
Composition per liter:

Peptone ........................................................... .23.0g

LiCl.................................................................. .15.0g

Agar .............................................................. ...10.0g

Mannitol......................................................... ...10.0g

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

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

Starch............................................................... ...1.0g

Esculin .............................................................. .0.8g

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

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

Phenol Red...................................................... ..0.08g

PALCAM selective supplement.................. .. 10.0mL

pH 7.2 ± 0.2 at 25oC

Source: This medium is available as a premixed powder from Oxoid Unipath.

PALCAM Selective Supplement: Composition per 10.0mL:

Ceftazidime................................................... .20.0mg

Polymyxin B................................................ ..10.0mg

Acriflavine*HCl ............................................ ..5.0mg

Preparation of PALCAM Selective Supple-
ment: Add components to distilled/deionized water
and bring volume to 10.0mL. Mix thoroughly. Filter
sterilize.

Egg Yolk Emulsion:

Composition:

Chicken egg yolks................................................ ..11

Whole chicken egg ................................................ ..1

Preparation of Egg Yolk Emulsion: Soak eggs with 1:100 dilution of saturated mercuric chloride so-
lution for 1 min. Crack eggs and separate yolks from whites. Mix egg yolks with 1 chicken egg.

Preparation of Medium: Add components, ex-
cept PALCAM selective supplement, to distilled/
deionized water and bring volume to 990.0mL. Mix
thoroughly. Gently heat and bring to boiling. Auto-
clave for 15 min at 15 psi pressure-121oC. Cool to
45o-50oC. Aseptically add sterile PALCAM selec-
tive supplement. Mix thoroughly. The addition of
25.0mL of egg yolk emulsion may aid in the recovery
of damaged Listeria. Pour into sterile Petri dishes or
distribute into sterile tubes.

Use: For the selective isolation, cultivation, and dif-

ferentiation of Listeria monocytogenes and other Listeria species.


Pasteurella haemolytica

Selective Medium

Composition per 1010.0mL:

Tryptose agar with peptic digest of blood......... . 1.0L

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

pH 7.2 ± 0.2 at 25oC

Tryptose Agar With Peptic Digest of Blood: Composition per liter:

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

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

Peptic digest of animal tissue........................... .10.0g

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

Glucose ........................................................... ...1.0g

Peptic digest of blood ................................ ..50.0mL

Preparation of Tryptose Agar With Peptic
Digest of Blood: Add components to distilled/
deionized water and bring volume to 950.0mL. Mix
thoroughly. Gently heat and bring to boiling. Auto-
clave for 15 min at 15 psi pressure-121oC. Cool to
45o-50oC. Aseptically add peptic digest of blood.
Mix thoroughly.

Antibiotic Solution:

Composition per 10.0mL:

Actidione (cycloheximide) .............................. ...0.1g

Novobiocin..................................................... .2.0mg

Neomycin..................................................... ...1.5mg

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: To 1.0L of cooled, ster-
ile tryptose agar with peptic digest of blood, asepti-
cally add 10.0mL of sterile antibiotic solution. Mix
thoroughly. Pour into sterile Petri dishes or distribute
into sterile tubes.

Use: For the selective cultivation of Pasteurella haemolytica.


Pasteurella multocida Selective Medium

Composition per 1020.0mL:

Tryptose agar with peptic digest of blood.. 1.0L
Antibiotic solution ... .................................. .10.0mL

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

pH 7.2 ± 0.2 at 25oC

Tryptose Agar With Peptic Digest of Blood: Composition per liter:

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

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

Peptic digest of animal tissue ........................ ...10.0g

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

Glucose ........................................................... ...1.0g

Peptic digest of blood ................................. .. 50.0mL

Preparation of Tryptose Agar with Peptic
Digest of Blood: Add components to distilled/
deionized water and bring volume to 950.0mL. Mix
thoroughly. Gently heat and bring to boiling. Auto-
clave for 15 min at 15 psi pressure-121oC. Cool to
45o-50oC. Aseptically add peptic digest of blood.
Mix thoroughly.

Antibiotic Solution:

Composition per 10.0mL:

Actidione (cycloheximide) .............................. ...0.1g

Novobiocin ..................................................... .0.01g

Erythrocin ..................................................... ..5.0mg

Preparation of Antibiotic Solution: Add com-
ponents to distilled/deionized water and bring vol-
ume to 10.0mL. Mix thoroughly. Filter sterilize.

K2TeO3 Solution:

Composition per 10.0mL:

K2TeO3............................................................ .5.0mg

Preparation of K2TeO3 Solution: Add K2TeO3 to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.

Caution: Potassium tellurite is toxic.

Preparation of Medium: To 1.0L of cooled, sterile tryptose agar with peptic digest of blood, aseptically add 10.0mL of sterile antibiotic solution and 10.0mL of ster-
ile K2TeO3 solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the selective cultivation of Pasteurella multocida.


PDM-114 Medium

Composition per liter:

Base solution............................................. .. 770.0mL

Bovine serum, heat inactivated................... . 100.0mL

Solution A................................................... .. 50.0mL

Solution B................................................... .. 50.0mL

Special 107 vitamin mix............................... . 30.0mL

pH 6.8 ± 0.2 at 25oC

Base Solution:

Composition per 770.0mL:

Pancreatic digest of casein............................... ..20.0g

NaCl.................................................................... 2.0g

L-Cysteine*HCl................................................ ...1.0g

Ascorbic acid ...................................................... 0.2g

Ammonium chloride....................................... ..0.16g

Adenine......................................................... .73.0mg

Guanosine .................................................. ...59.0mg

Adenosine 5ด-monophosphate .................... ..52.0mg

Uracil ........................................................... .37.0mg

Cytosine ..................................................... .. 30.0mg

Ferric ammonium citrate.............................. ..22.8mg

Adenosine 5ด-diphosphate .......................... ..16.6mg

Adenosine 5ด-triphosphate.............................. .7.6mg

Preparation of Base Solution: Add components to distilled/deionized water and bring volume to 770.0mL. Mix thoroughly. Adjust pH to 6.8 with NaOH. Autoclave for 15 min at 15 psi pressure-
121oC. Cool to room temperature.

Solution A:

Composition per 50.0.0mL:

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

Preparation of Solution A: Add glucose to dis-
tilled/deionized water and bring volume to 50.0mL. Mix thoroughly. Filter sterilize.

Solution B:

Composition per 50.0.0mL:

K2HPO4........................................................... ...1.0g

KH2PO4........................................................... ...0.6g

Preparation of Solution B: Add components to distilled/deionized water and bring volume to 50.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pres-
sure-121oC. Cool to room temperature.

Special 107 Vitamin Mix:

Composition per 120.0.0mL:

Solution 4 vitamins.................................... .100.0mL

Solution 2..................................................... ..1.2mL

Solution 1..................................................... ..0.4mL

Solution 3..................................................... ..0.4mL

Solution 1:

Composition per 100.0.0mL:

Absolute ethanol ...................................... ..100.0mL

DL-6,8-Thioctic acid, oxidized .................. ..100.0mg

Preparation of Solution 1: Add DL-6,8-thioctic acid  to  absolute  ethanol  and  bring  volume  to 100.0mL. Mix thoroughly. Filter sterilize.

Solution 2:

Vitamin B12 .................................................. .40.0mg

Preparation of Solution 2: Add vitamin B12 to distilled/deionized  water  and  bring  volume  to 100.0mL. Mix thoroughly. Filter sterilize.

Solution 3:

Tween 80........................................................ ..50.0g

Absolute ethanol ...................................... ..100.0mL

Preparation of Solution 3: Add Tween 80 to ab-
solute ethanol and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

Use: For the cultivation and differentiation of microor-

ganisms based on their ability to produce H2S. Microor-
ganisms that produce H2S turn the medium black.


Peptone Meat Extract Soil Extract Agar

(PFE Agar)

Composition per liter:

Agar .............................................................. ...12.0g

Proteose peptone no. 3....................................... .5.0g

Meat extract ..................................................... ..3.0g

Tap water.................................................. ..850.0mL

Soil extract................................................ ..150.0mL

Glycerol ..................................................... ..20.0mL

pH 7.0 ± 0.2 at 25oC

Soil Extract:

Composition per liter:

Garden soil, air dried ................................... ..400.0g

Preparation of Soil Extract: Pass 400.0g of air-
dried garden soil through a coarse sieve. Add soil to
960.0mL of tap water. Mix thoroughly. Autoclave for
60 min at 15 psi pressure-121oC. Cool to room tem-
perature. Allow residue to settle. Decant supernatant
solution. Filter through Whatman filter paper. Dis-
tribute into bottles in 200.0mL volumes. Autoclave
for 15 min at 15 psi pressure-121oC. Store at room
temperature until clear.

Preparation of Medium: Add components to tap 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 dishes or leave in tubes. Use: For the cultivation and maintenance of Mycobac-
terium avium, Mycobacterium gastri, Mycobacterium kansasii, Mycobacterium marinum, Mycobacterium scrofulaceum, and Mycobacterium terrae.


Peptone Starch Dextrose Agar

(PSD Agar)

(Dunkelberg Agar)

Composition per liter:

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

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

Soluble starch.................................................. .10.0g

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

Na2HPO4........................................................... .1.0g

NaH2PO4........................................................... .1.0g

pH 6.8 ± 0.2 at 25oC

Preparation of Medium: Add starch to approxi-
mately 100.0mL of cold distilled/deionized water.
Mix thoroughly. Add starch solution to 400.0mL of
boiling  distilled/deionized  water.  Add  remaining
components. Mix thoroughly. Bring volume to 1.0L

with distilled/deionized water. Autoclave for 12 min

at 8 psi pressure-112oC. Pour into sterile Petri dishes or distribute into screw-capped tubes.

Use: For the selective isolation and cultivation of Gardnerella vaginalis.


Peptone Water

Composition per liter:

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

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

pH 7.2 ± 0.2 at 25oC

Source: This medium is available as a premixed
powder from  BD Diagnostic Systems and Oxoid Un-
ipath.

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 nonfastidious microor-
ganisms, for carbohydrate fermentation tests, and for performing the indole test.


Peptone Water

with Andrade’s Indicator

Composition per liter:

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

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

Andrade’s indicator.................................... . 100.0mL

Carbohydrate solution................................. .. 20.0mL

pH 7.4 ± 0.2 at 25oC

Source: This medium is available as a premixed powder from Oxoid Unipath.

Andrade’s Indicator:

Composition per 100.0mL:

NaOH (1N solution) ................................... . 16.0mL

Acid Fuchsin................................................... ...0.1 g

Caution: Acid Fuchsin is a potential carcinogen and care must be taken to avoid inhalation of the powdered dye and contact with the skin.

Carbohydrate Solution:

Composition per 20.0mL:

Carbohydrate............................................. .5.0-10.0g

Preparation of Carbohydrate Solution: Add
carbohydrate to distilled/deionized water and bring
volume to 20.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, ex-
cept carbohydrate solution, to distilled/deionized wa-
ter and bring volume to 980.0mL. Mix thoroughly.
Adjust pH to 7.4 if necessary. Distribute into tubes
containing an inverted Durham tube. Fill each tube

with 9.8mL of medium. Autoclave for 15 min at 15

psi pressure-121oC. Aseptically add 0.2mL of sterile carbohydrate solution to each tube.

Use: For use in carbohydrate fermentation tests. Fer-
mentation is determined by the production of acid— broth turns pink—and formation of gas—bubble trapped in Durham tube.


Peptone Yeast Trypticase Agar

(ATCC Medium 118)

Composition per liter:

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

Peptone.............................................................. .6.0g

Trypticase (pancreatic digest of casein)............. ..4.0g

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

Beef extract...................................................... ...1.5g

Glucose ........................................................... ...1.0g

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 of a variety of heterotrophic bacteria.


Petragnani Medium

Composition per 2398.0mL:

Skim milk..................................................... ..100.0g

Potato flour ..................................................... .36.4g

L-Asparagine ..................................................... .5.1g

Pancreatic digest of casein................................. .5.1g

Malachite Green................................................ ..1.2g

Whole egg................................................ .1277.0mL

Egg yolk.................................................. ...121.0mL

Glycerol ..................................................... ..60.0mL

pH 7.0 ± 0.2 at 25oC

Source: This medium is available as a prepared me-
dium from  BD Diagnostic Systems.

Preparation of Medium: Add components—ex-
cept whole egg, egg yolk, and glycerol—to distilled/
deionized water and bring volume to 940.0mL. Mix
thoroughly. Add glycerol. Gently heat while stirring
and bring to boiling. Autoclave for 15 min at 15 psi
pressure-121oC. Cool to 45o-50oC. Scrub the egg-
shells with soap. Let stand in a soap solution for 30
min. Rinse in running water. Soak eggs in 70% etha-
nol for 15 min. Break the eggs into a sterile container.
Homogenize by shaking. Filter through four layers of
sterile cheesecloth into a sterile graduated cylinder.
Measure out 1277.0mL. Add separated egg yolks to


 

another sterile container. Measure out 121.0mL.

Aseptically add homogenized whole egg and egg yolk to cooled sterile basal medium. Mix thoroughly. Aseptically distribute into sterile tubes. Inspissate at 85o-90oC (moist heat) for 45 min.

Use: For the isolation and cultivation of Mycobacte-
rium species from clinical specimens.


Petragnani Medium

Composition per 2285.0mL:

Potato............................................................ ..500.0g

Potato flour ..................................................... .36.0g

Malachite Green................................................ ..1.2g

Whole egg................................................ . 1200.0mL

Whole milk ................................................ . 900.0mL

Egg yolk................................................... ... 115.0mL

Glycerol ..................................................... .. 70.0mL

pH 7.2 ± 0.2 at 25oC

Source: This medium is available as a prepared me-
dium from  BD Diagnostic Systems.

Preparation of Medium: Peel and dice potato.
Add potato to 500.0mL of distilled/deionized water.
Gently heat and bring to boiling. Continue boiling for
30 min. Filter solids through two layers of cheese-
cloth. Combine potato solids with remaining compo-
nents, except whole egg, egg yolk, and glycerol. Mix
thoroughly. Add glycerol. Gently heat while stirring
and bring to boiling. Autoclave for 15 min at 15 psi
pressure-121oC. Cool to 45o-50oC. Scrub the egg-
shells with soap. Let stand in a soap solution for 30
min. Rinse in running water. Soak eggs in 70% etha-
nol for 15 min. Break the eggs into a sterile container.
Homogenize by shaking. Filter through four layers of
sterile cheesecloth into a sterile graduated cylinder.
Measure out 1200.0mL. Add separated egg yolks to
another sterile container. Measure out 115.0mL.

Aseptically add homogenized whole egg and egg yolk to cooled sterile basal medium. Mix thoroughly. Aseptically distribute into sterile tubes. Inspissate at 85o-90oC (moist heat) for 45 min.

Use: For the isolation and cultivation of Mycobacteri-
um species from clinical specimens.


Pfizer Selective Enterococcus Agar

(PSE Agar)

Peptone C......................................................... .17.0g

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

Bile.................................................................. ..10.0g

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

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

Peptone B......................................................... ...3.0g

Esculin .............................................................. .1.0g

Sodium citrate................................................... ..1.0g

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

NaN3 .............................................................. ..0.25g

pH 7.1 ± 0.2 at 25oC

Caution: Sodium azide is toxic. Azides also react
with metals and disposal must be highly diluted.
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 selective isolation, cultivation, and enu-
meration of Enterococcus species by the multiple tube technique.


PGA

(Potato Glucose Agar)

Composition per liter:

Potatoes........................................................ ..500.0g

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

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

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

Preparation of Medium: Slice potatoes with
skin. Place 500.0g of potatoes in 1.0L of distilled/
deionized water. Gently heat and bring to boiling. Al-
low to boil for 20 min. Filter through Whatman filter
paper. Add agar and other components to filtrate.
Bring volume to 1.0L with distilled/deionized water.
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 of numerous filamentous
fungi.


PGP Broth

(Peptone Glycerol Phosphate Broth)

Composition per liter:

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

K2HPO4........................................................... ...2.0g

Glycerol ..................................................... ..10.0mL

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 and maintenance of Serratia marcescens.


PGT Medium

Composition per liter:

Casamino acids ............................................... .30.0g

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

MgSO4*7H2O.................................................. .0.45g

Maltose ........................................................... ...0.2g

L-Cystine............................................................ .0.2g

DL-Tryptophan.................................................. ..0.1g

Solution 3................................................... . 100.0mL

Solution 2...................................................... .. 2.0mL

Calcium pantothenate (0.1% solution) .......... .. 0.5mL

pH 6.8 ± 0.2 at 25oC

Solution 3:

Composition per 500.0mL:

Maltose ........................................................ ..200.0g

CaCl2.................................................................. .1.5g

Calcium pantothenate (0.1% solution) .......... .. 3.0mL

FeSO4 (1% in 1N HCl)................................. .. 0.2mL

Preparation of Solution 3: Add components to distilled/deionized  water  and  bring  volume  to 500.0mL. Mix thoroughly. Autoclave for 15 min at 7 psi pressure-111oC. Cool to 45o-50oC.

Solution 2:

Composition per 100.0mL:

β-Alanine...................................................... ..0.115g

Nicotinic acid................................................ ..0.115g

CuSO4*5H2O.................................................. ..0.05g

ZnSO4*7H2O............................................... ...0.045g

MnCl2*4H2O ............................................... ...0.015g

Pimelic acid ................................................... ..7.5mg

HCl, concentrated .......................................... . 3.0mL

Preparation of Solution 2: Add components to distilled/deionized  water  and  bring  volume  to 100.0mL. Mix thoroughly.

Preparation of Medium: Add components, except
solution 3, to distilled/deionized water and bring vol-
ume to 900.0mL. Mix thoroughly. Adjust pH to 6.8 with
50% KOH. Gently heat and bring to boiling. Autoclave
for 15 min at 15 psi pressure-121oC. Cool to 45o-50oC.
Aseptically add sterile solution 3. Mix thoroughly.

Aseptically distribute into sterile tubes or flasks. Use: For the cultivation and maintenance of Coryne-
bacterium diphtheriae.

Phenol Red Agar

Composition per liter:

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

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

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

Phenol Red................................................... ...0.018g

Carbohydrate solution................................. .. 20.0mL

pH 7.4 ± 0.2 at 25oC

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

 Carbohydrate Solution:

Composition per 20.0mL:

Carbohydrate............................................ .5.0-10.0g

Preparation of Carbohydrate Solution: Add
carbohydrate to distilled/deionized water and bring
volume to 20.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium: Add components, ex-
cept carbohydrate solution, to distilled/deionized wa-
ter and bring volume to 980.0mL. Mix thoroughly.
Adjust pH to 7.4 if necessary. Autoclave for 15 min
at 15 psi pressure-121oC. Cool to 45o-50oC. Asepti-
cally add 20.0mL of sterile carbohydrate solution.
Pour into sterile Petri dishes or distribute into sterile
tubes. Allow tubes to cool in a slanted position.

Use: For the determination of fermentation reac-
tions. Bacteria that can ferment the added carbohy-
drate turn the medium yellow.


Phenol Red Agar

Composition per liter:

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

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

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

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

Phenol Red.................................................. ...0.025g

Carbohydrate solution................................. ..20.0mL

pH 7.4 ± 0.2 at 25oC

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

Carbohydrate Solution:

Composition per 20.0mL:

Carbohydrate............................................ .5.0-10.0g

Preparation of Carbohydrate Solution: Add
carbohydrate to distilled/deionized water and bring
volume to 20.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, ex-
cept carbohydrate solution, to distilled/deionized wa-
ter and bring volume to 980.0mL. Mix thoroughly.
Adjust pH to 7.4 if necessary. Autoclave for 15 min
at 15 psi pressure-121oC. Cool to 45o-50oC. Asepti-
cally add 20.0mL of sterile carbohydrate solution.
Pour into sterile Petri dishes or distribute into sterile
tubes. Allow tubes to cool in a slanted position.

Use: For the determination of fermentation reac-
tions. Bacteria that can ferment the added carbohy-
drate turn the medium yellow.