รวมอาหารเลี้ยงเชื้อหมวด O

posted on 04 Jun 2012 22:29 by moomsabuy in medium directory Knowledge

O157:H7(+) Plating Medium

Composition per liter:

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

Sorbitol........................................................... ..12.0g

Salicin ........................................................... ...10.0g

Inositol............................................................ ..10.0g

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

Adonitol............................................................ ..8.0g

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

Tryptone............................................................ ..5.0g

Proteose peptone................................................ .3.0g

Bile salts no. 3................................................ ...1.25g

Indoxyl-β-D-galactopyranoside ..................... ...0.12g

5-Bromo-4-chloro-3-indoxyl-

β-D-galactopyranoside.............................. ..0.12g

Phenol Red......................................................... .0.1g

Isopropyl-β-D-thiogalactopyranoside................ ..0.1g

Novobiocin solution....................................... . 1.0mL

Tellurite solution.......................................... ... 1.0mL

pH 6.8 ± 0.2 at 25oC

Source: This medium is available as a premixed powder from BIOSYNTH International, Inc.

Novobiocin Solution:

Composition per 10.0mL:

Novobiocin ..................................................... ...0.1g

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

Tellurite Solution:

Composition per 10.0mL:

K2TeO3............................................................ ..0.01g

Preparation of 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 novobiocin solution and tellurite solution, to dis-
tilled/deionized water and bring volume to 998.0mL.
Mix thoroughly. Gently heat while stirring and bring
to boiling. Autoclave for 15 min at 15 psi pressure-
121oC. Cool to 50oC. Aseptially add 1.0mL sterile
novobiocin solution and 1.0 sterile tellurite solution.
Mix thoroughly. Pour into sterile Petri dishes.
Use: For the detection of Escherichia coli O157:H7.
E.coli O157:H7 grow with blue-black colonies and
E.coli non-O157 with green-yellow colonies.


O157:H7 ID Agar

Composition per liter:

Proprietary

Source: This medium is available from bioM้rieux.

Use: A new chromogenic medium for the detection of Escherichia coli O157:H7.


Oatmeal Agar

Composition per 750.0mL:

Oatmeal, instant for babies .............................. .40.0g

Agar ................................................................. ..5.0g

Preparation of Medium: Add agar to distilled/
deionized water and bring volume to 500.0mL. Mix
thoroughly. Gently heat and bring to boiling. Add in-
stant oatmeal for babies to distilled/deionized water
and bring volume to 250.0mL. Mix thoroughly. Add
the oatmeal solution to the melted agar solution. Mix
thoroughly. Autoclave for 15 min at 15 psi pressure-
121oC. Pour into sterile Petri dishes or aseptically
distribute into sterile tubes.

Use: For the cultivation of fungi.


Oatmeal Agar

(ATCC 551)

Composition per liter:

Oatmeal............................................................ .60.0g

Agar .............................................................. ...12.5g

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.
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 cultivation of fungi and actinomycetes, par-
ticularly for macrospore formation.


Oatmeal Agar

(OA)

Composition per liter:

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

Oatmeal, rolled oats....................................... ...10.0g

Glycerin ........................................................ .5.0mL

Lactic acid...................................................... .0.2mL

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 many filamentous fungi.


Ogawa TB Medium

Composition per 300.0mL:

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

Homogenized whole egg .......................... .200.0mL

Glycerol ........................................................ . 6.0mL

Malachite Green (2% solution)..................... .. 6.0mL

pH 6.5 ± 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: Add components, ex-
cept homogenized whole egg, to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure-121oC. Cool
to 45o-50oC. Aseptically add 200.0mL of sterile ho-
mogenized whole egg. Mix thoroughly. Aseptically
distribute into sterile screw-capped tubes in 7.0mL
volumes. Inspissate at 85o-90oC (moist heat) for 60
min.

Use: For the isolation and cultivation of Mycobacte-
rium species, except for Mycobacterium leprae.


Oleic Albumin Complex

Composition per liter:

Bovine albumin fraction V .............................. .50.0g

NaCl.................................................................. ..8.5g

Oleic acid...................................................... .. 0.6mL

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

Use: For use in media employed for the cultivation of mycobacteria.


 Salmonella Agar

Composition per liter:

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

Sucrose........................................................... ..13.0g

Lactose............................................................ ..11.5g

Trisodium citrate-5,5-hydrate............................. .9.3g

Meat peptone ................................................... ...6.8g

Beef extract...................................................... ...6.0g

L-Phenylalanine ................................................ ..5.0g

Na2S2O3*5H2O............................................... ..4.25g

Bile salt mixture............................................. .3.825g

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

Na2HPO4*2H2O............................................... ...1.0g

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

Metachrome Yellow....................................... ...0.47g

MgSO4*7H2O.................................................. ...0.4g

Aniline Blue................................................... ...0.25g

Neutral Red................................................... ..0.022g

Brilliant Green ......................................... ..0.00166g

pH 7.1 ± 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 isolation and cultivation of Salmonella from feces.


ONPG Broth

Composition per liter:

Peptone water............................................ .750.0mL

ONPG solution......................................... ..250.0mL

pH 7.2-7.4 at 25oC

ONPG Solution:

Composition per 250.0mL:

ONPG (o-Nitrophenyl-β-

D-galactopyranoside) ................................ ...1.5g

Sodium phosphate

buffer (0.01M, pH 7.5) ....................... .250.0mL

Preparation of ONPG Solution: Add ONPG to 250.0mL of sodium phosphate buffer. Mix thorough-
ly. Filter sterilize.

Peptone Water:

Composition per 750.0mL:

Peptone.............................................................. .7.5g

NaCl.............................................................. ...3.75g

Preparation of Peptone Water: Add compo-
nents to distilled/deionized water and bring volume
to 750.0mL. Mix thoroughly. Gently heat and bring
to boiling. Adjust pH to 8.0-8.4. Continue boiling for
10 min. Filter through Whatman no. 1 filter paper.
Readjust pH of filtrate to 7.2-7.4. Autoclave for 20
min at 10 psi pressure-115oC. Cool to 25oC.
Preparation of Medium: Aseptically combine
the sterile ONPG solution with the cooled, sterile
peptone water. Mix thoroughly. Aseptically distrib-
ute into tubes in 2.5-3.0mL volumes. Store at 4oC for
up to 1 month.

Use: For the differentiation of a variety of Gram-
negative bacteria based on production of β-galactosi-
dase. For the differentiation of lactose-delayed bacte-
ria from lactose-negative bacteria. For the differenti-
ation  of  Pseudomonas  cepacia (positive)  and

Pseudomonas  maltophila (positive)  from  other

Pseudomonas species (negative). Bacteria that pro-
duce β-galactosidase turn the medium yellow.


OR Indicator Agar

(Oxidation-Reduction Indicator Agar)

Composition per liter:

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

Sodium glycerol phosphate.............................. .10.0g

Sodium thioglycolate........................................ ...1.7g

CaCl2*2H2O ..................................................... ..0.1g

Methylene Blue............................................. ...6.0mg

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 use as an indicator of oxygen-free condi-
tions in anaerobic culture chambers.


Oral Fusobacterium Medium

 Composition per liter:

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

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

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

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

Beef extract...................................................... ...3.0g

Soluble starch................................................... ...2.0g

NaNO3 .............................................................. .1.0g

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

L-Cysteine*HCl*H2O......................................... .0.5g

Ethyl Violet solution.................................... . 10.0mL

Bacitracin solution........................................ . 10.0mL

pH 7.6 ± 0.2 at 25oC

Ethyl Violet Solution:

Composition per 10.0mL:

Ethyl Violet...................................................... .0.04g

Preparation of Ethyl Violet Solution: Add Eth-
yl Violet to distilled/deionized water and bring vol-
ume to 10.0mL. Mix thoroughly. Filter sterilize.

Bacitracin Solution:

Composition per 10.0mL:

Bacitracin...................................................... ...1.0mg

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

Preparation of Medium: Add components, ex-
cept Ethyl Violet solution and bacitracin solution, to
distilled/deionized  water  and  bring  volume  to
980.0mL. Mix thoroughly. Gently heat and bring to
boiling. Autoclave for 15 min at 15 psi pressure-
121oC. Cool to 45o-50oC. Aseptically add sterile
Ethyl Violet solution and bacitracin solution. Mix

thoroughly. Pour into sterile Petri dishes or distribute

into sterile tubes.

Use: For the selective isolation and cultivation of oral Fusobacterium species, especially Fusobacteri-
um nucleatum.


Oral Treponema Medium

Composition per 1250.0mL:

Veal heart, fresh ground................................ ..1.0Kg

Thiopeptone .................................................. ...20.0g

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

Ionagar no. 2 ..................................................... .2.0g

Glutathione (1% solution)........................ ...100.0mL

Rabbit serum or ascitic fluid..................... ..100.0mL

Eggs, whole fresh.................................................. ..2

pH 6.8-7.0 at 25oC

Preparation of Medium: Add agar to 50.0mL of
distilled/deionized water. Gently heat and bring to
boiling. Autoclave for 15 min at 15 psi pressure-
121oC. Cool to 45o-50oC. In a separate flask, add
finely ground veal heart to 1.0L of distilled/deionized
water.  Add  remaining  components,  except  glu-
tathione, rabbit serum, and agar. Gently heat and
bring to 70oC. Adjust pH to 7.4. Gently heat and
bring to 100oC. Continue heating at 100oC for 2h.
Skim off fat. Filter through glass wool. Adjust pH to
7.6. Gently heat and bring to 100oC. Maintain at
100oC for 30 min. Store at 4oC for 18h. Sterilize in an
Arnold sterilizer for 30 min at 100oC on two consec-
utive days. Cool to 45o-50oC. Aseptically add rabbit
serum, glutathione solution, and sterile cooled agar
solution. Mix thoroughly. Aseptically distribute into
sterile tubes or flasks.

Use: For the isolation and cultivation of Treponema denticola and Treponema oralis.


Organic Acid Medium KP

(Organic Acid Medium, Kauffmann and Petersen)

Composition per liter:

Gelatin........................................................... ...10.0g

Bromthymol Blue ......................................... .0.024g

Organic acid solution................................. .100.0mL

pH 7.4 ± 0.2 at 25oC

Organic Acid Solution:

Composition per 100.0mL:

Organic acid................................................... ...10.0g

Preparation of Organic Acid Solution: Add
organic acid to distilled/deionized water and bring
volume to 100.0mL. Sodium potassium D-tartrate,
sodium citrate, or mucic acid may be used. Mix thor-
oughly.

Preparation of Medium: Add components, ex-
cept organic acid solution, 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 organic acid solution. Mix thoroughly.
Aseptically distribute into sterile tubes or flasks.

Use: For the cultivation and differentiation of mem-
bers of the Enterobacteriaceae based on their ability to utilize different organic acids as carbon source. Bacteria that utilize tartrate, citrate, or mucate turn the medium yellow.


Ornithine Broth

Composition per liter:

L-Ornithine......................................................... .5.0g

Peptone or gelysate.......................................... ...5.0g

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

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

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

pH 6.5 ± 0.2 at 25oC

Preparation of Medium: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH so that is will be 6.5 ± 0.2 after sterilization. Distribute into 16 ื 150mm screw-capped tubes in 5.0mL volumes. Autoclave medium with

loosely capped tubes for 10 min at 15 psi pressure-
121oC. Screw the caps on tightly for storage and after inoculation.

Use: For the cultivation and differentiation of bacte-
ria based on their ability to decarboxylate the amino acid ornithine. Bacteria that decarboxylate ornithine turn the medium turbid purple.


Ornithine Broth with NaCl

Composition per liter:

L-Ornithine......................................................... .5.0g

Peptone or gelysate.......................................... ...5.0g

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

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

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

pH 6.5 ± 0.2 at 25oC

Preparation of Medium: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH so that is will be 6.5 ± 0.2 after sterilization. Distribute into 16 ื 150mm screw-capped tubes in 5.0mL volumes. Autoclave medium with

loosely capped tubes for 10 min at 15 psi pressure-
121oC. Screw the caps on tightly for storage and after inoculation.

Use: For the cultivation and differentiation of Vibrio spp. based on their ability to decarboxylate the amino acid ornithine. Bacteria that decarboxylate ornithine turn the medium turbid purple.


Oxford Agar

(Listeria Selective Agar, Oxford)

Composition per liter:

Special peptone................................................ .23.0g

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

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

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

Cornstarch........................................................ ..1.0g

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

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

Antibiotic inhibitor .................................... ...10.0mL

pH 7.0 ± 0.2 at 25oC

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

Antibiotic Inhibitor:

Composition per 10.0mL:

Cycloheximide................................................... .0.4g

Colistin sulfate................................................ ..0.02g

Fosfomycin ..................................................... .0.01g

Acriflavine ..................................................... .5.0mg

Cefotetan......................................................... .2.0mg

Ethanol (50% solution).............................. ...10.0mL

Preparation of Antibiotic Inhibitor: Add anti-
biotics to 10.0mL of ethanol. Mix thoroughly. Filter
sterilize.

Preparation of Medium: Add components, ex-
cept antibiotic inhibitor, to distilled/deionized water
and bring volume to 990.0mL. Mix thoroughly. Gen-
tly heat and bring to boiling. Autoclave for 15 min at
15 psi pressure-121oC. Cool to 45o-50oC. Aseptical-
ly add 10.0mL of sterile antibiotic inhibitor. Mix
thoroughly. Pour into sterile Petri dishes or distribute
into sterile tubes.

Use: For the isolation and cultivation of Listeria monocytogenes from specimens containing a mixed bacterial flora.


Oxford Agar, Modified

(Listeria Selective Agar,Modified Oxford)

(MOX Agar)

Composition per liter:

Special peptone................................................ .23.0g

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

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

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

Cornstarch........................................................ ..1.0g

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

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

Antibiotic inhibitor .................................... ...10.0mL

pH 7.0 ± 0.2 at 25oC

 Antibiotic Inhibitor:

Composition per 10.0mL:

Moxalactam ................................................... .0.015g

Colistin sulfate................................................ ..0.01g

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

Preparation of Medium: Add components, ex-
cept antibiotic inhibitor, to distilled/deionized water
and bring volume to 990.0mL. Mix thoroughly. Gen-
tly heat and bring to boiling. Autoclave for 10 min at
15 psi pressure-121oC. Cool to 45o-50oC. Aseptical-
ly add 10.0mL of sterile antibiotic inhibitor. Mix
thoroughly. Pour into sterile Petri dishes or distribute
into sterile tubes.

Use: For the isolation and cultivation of Listeria monocytogenes from specimens containing a mixed bacterial flora.


Oxidation-Fermentation Medium

(OF Medium)

Composition per liter:

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

Agar ................................................................. ..2.5g

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

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

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

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

pH 6.8 ± 0.1 at 25oC

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

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 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 100.0mL of sterile carbohydrate so-
lution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use:  For  differentiating  Gram-negative  bacteria based upon determining the oxidative and fermenta-
tive metabolism of carbohydrates.


Oxidation-Fermentation

Medium, Hugh-Leifson’s

(Hugh-Leifson’s Oxidation

Fermentation Medium)

Composition per liter:

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

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

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

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

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

Bromthymol Blue solution (0.2% )............ ...15.0mL

pH 7.1 ± 0.2 at 25oC

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 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 100.0mL of sterile carbohydrate so-
lution. Mix thoroughly. Pour into sterile Petri dishes
or distribute into sterile tubes.

Use: For differentiating Gram-negative bacteria, such as Vibrio species, based upon determining the oxidative and fermentative metabolism of carbohy-
drates. Bacteria that ferment the carbohydrate turn the medium yellow.


Oxidation-Fermentation

Medium, King’s

(King’s OF Medium)

Composition per liter:

Base solution............................................ ..900.0mL

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

pH to 7.3 ± 0.2

Base Solution:

Composition per liter:

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

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

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

Phenol Red (1.5% solution)........................... .2.0mL

Preparation of Base Solution: Add carbohy-
drate to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

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 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 100.0mL of sterile carbohydrate so-
lution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For differentiating bacteria based upon deter-
mining the oxidative and fermentative metabolism of carbohydrates. Bacteria that ferment the carbohy-
drate turn the medium yellow.


Oxidative-Fermentative Medium

(OF Medium)

Composition per liter:

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

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

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

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

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

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

pH 6.8 ± 0.1 at 25oC

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

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 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 100.0mL of sterile carbohydrate so-
lution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For differentiating bacteria based upon deter-
mining the oxidative and fermentative metabolism of carbohydrates. Bacteria that ferment the carbohy-
drate turn the medium yellow.