Handbook of Microbiological Media, Fourth Edition part 192 pdf

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L.. Preparation of Medium: Add components, except defibrinated blood, to distilled/deionized wat

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Wilkins-Chalgren Anaerobe Agar with GN Supplement 1905

K2HPO4 0.5g

MgSO4·7H2O 0.25g

Na2SO3, anhydrous 0.05g

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized

water and bring volume to 1.0L Mix thoroughly Gently heat and bring

to boiling Distribute into tubes or flasks Autoclave for 15 min at 15

psi pressure–121°C Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation and maintenance of Xanthomonas albilineans

and other Xanthomonas species.

Wilkins-Chalgren Agar Compositionper liter:

Agar 15.0g

Gelatin peptone 10.0g

Pancreatic digest of casein 10.0g

NaCl 5.0g

Yeast extract 5.0g

Glucose 1.0g

L-Arginine 1.0g

Sodium pyruvate 1.0g

Hemin 5.0mg

Vitamin K1 (menadione) 0.5mg

pH 7.1 ± 0.2 at 25°C

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

Di-agnostic Systems

psi pressure–121°C Cool to 50°–55°C Add antibiotic to be assayed;

varying concentrations of antibiotics are used Mix thoroughly Pour

into sterile Petri dishes or leave in tubes

Use: For the cultivation and maintenance of anaerobic bacteria For

standardized antimicrobic susceptibility testing to determine the

mini-mum inhibitory concentrations of antimicrobics for anaerobic bacteria

Wilkins-Chalgren Anaerobe Agar

Compositionper liter:

Agar 10.0g

NaCl 5.0g

Yeast extract 5.0g

Glucose 1.0g

L-Arginine 1.0g

Hemin 5.0mg

Menadione 0.5mg

Defibrinated blood 50.0mL

Tween™ 80 1.0mL

pH 7.1 ± 0.2 at 25°C

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

Unipath

Preparation of Medium: Add components, except defibrinated

blood, to distilled/deionized water and bring volume to 950.0mL Mix

thoroughly Gently heat and bring to boiling Distribute into tubes or

flasks Autoclave for 15 min at 15 psi pressure–121°C Cool to 50°–

55°C Aseptically add 50.0mL of defibrinated blood Mix thoroughly Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation of nonsporulating anaerobes

Wilkins-Chalgren Anaerobe Agar with Cysteine Compositionper liter:

Agar 10.0g Pancreatic digest of casein 10.0g Gelatin peptone 10.0g NaCl 5.0g Yeast extract 5.0g Glucose 1.0g

L-Arginine 1.0g Sodium pyruvate 1.0g

L-Cysteine·HCl 0.3g Hemin 5.0mg Menadione 0.5mg Defibrinated blood 50.0mL Tween™ 80 1.0mL

pH 7.1 ± 0.2 at 25°C

Preparation of Medium: Add components, except defibrinated blood, to distilled/deionized water and bring volume to 950.0mL Mix thoroughly Gently heat and bring to boiling Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C Cool to 50°– 55°C Aseptically add 50.0mL of defibrinated blood Mix thoroughly Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation of Agromonas oligotrophica, Falcivibrio gran-dis, and Falcivibrio vaginalis.

Wilkins-Chalgren Anaerobe Agar with GN Supplement Compositionper liter:

Agar 10.0g Pancreatic digest of casein 10.0g Gelatin peptone 10.0g NaCl 5.0g Yeast extract 5.0g Glucose 1.0g

L-Arginine 1.0g Sodium pyruvate 1.0g Hemin 5.0mg Menadione 0.5mg Defibrinated blood 50.0mL

GN anaerobe selective supplement 20.0mL

pH 7.1 ± 0.2 at 25°C

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

GN Anaerobe Selective Supplement Compositionper 20.0mL:

Nalidixic acid 10.0mg Hemin 5.0mg Sodium succinate 2.5mg Vancomycin 2.5mg Menadione 0.5mg

Preparation of GN Anaerobe Selective Supplement: Add components to distilled/deionized water and bring volume to 20.0mL Mix thoroughly Filter sterilize

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1906 Wilkins-Chalgren Anaerobe Agar with NS Supplement

blood and GN anaerobe selective supplement, to distilled/deionized

water and bring volume to 900.0mL Mix thoroughly Gently heat and

bring to boiling Distribute into tubes or flasks Autoclave for 15 min

at 15 psi pressure–121°C Cool to 50°–55°C Aseptically add 20.0mL

of GN anaerobe selective supplement and 50.0mL of defibrinated

blood Bring volume to 1.0L with distilled/deionized water Mix

thor-oughly Pour into sterile Petri dishes or leave in tubes

Use: For the selective isolation of Gram-negative anaerobes

Wilkins-Chalgren Anaerobe Agar

with NS Supplement Compositionper liter:

Agar 10.0g

NaCl 5.0g

Yeast extract 5.0g

Glucose 1.0g

L-Arginine 1.0g

Hemin 5.0mg

Menadione 0.5mg

NS anaerobe selective supplement 20.0 mL

Tween™ 80 1.0mL

pH 7.1 ± 0.2 at 25°C

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

Unipath

NS Anaerobe Selective Supplement:

Compositionper 20.0mL:

Nalidixic acid 0.01g

Hemin 5.0mg

Menadione 0.5mg

Preparation of NS Anaerobe Selective Supplement: Add

components to distilled/deionized water and bring volume to 20.0mL

Mix thoroughly Filter sterilize

blood and NS anaerobe selective supplement, to distilled/deionized

water and bring volume to 900.0mL Mix thoroughly Gently heat and

bring to boiling Distribute into tubes or flasks Autoclave for 15 min

at 15 psi pressure–121°C Cool to 50°–55°C Aseptically add 20.0mL

of NS anaerobe selective supplement and 50.0mL of defibrinated

blood Bring volume to 1.0L with distilled/deionized water Mix

thor-oughly Pour into sterile Petri dishes or leave in tubes

Use: For the selective isolation of nonsporulating anaerobes

Wilkins-Chalgren Anaerobe Broth

(Anaerobe Broth, MIC) Compositionper liter:

NaCl 5.0g

Yeast extract 5.0g

Glucose 1.0g

L-Arginine 1.0g

Hemin 5.0mg Menadione 0.5mg

pH 7.1 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from BD Di-agnostic Systems and Oxoid Unipath

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Distribute into tubes

or flasks Autoclave for 15 min at 15 psi pressure–121°C

Use: For the cultivation and antimicrobial susceptibility (MIC) testing

of anaerobic bacteria

Wilkins-Chalgren Anaerobe Medium

with Yeast Extract Compositionper liter:

Yeast extract 20.0g Pancreatic digest of casein 10.0g Gelatin peptone 10.0g NaCl 5.0g Yeast extract 5.0g Glucose 1.0g

L-Arginine 1.0g Sodium pyruvate 1.0g Hemin 5.0mg Menadione 0.5mg

pH 7.1 ± 0.2 at 25°C

Use: For the cultivation of Selenomonas acidaminovorans.

Wilkins-Chalgren Anaerobic HiVeg Agar Base

with Blood Compositionper liter:

Agar 10.0g Plant hydrolysate 10.0g Plant peptone 10.0g NaCl 5.0g Yeast extract 5.0g

L-Arginine 1.0g Glucose 1.0g Sodium pyruvate 1.0g

Fe4(P2O7)3·H2O 5.0mg Menadione 0.5mg Defibrinated blood 50.0mL

pH 7.1 ± 0.2 at 25°C

Source: This medium, without blood, is available as a premixed pow-der from HiMedia

Preparation of Medium: Add components, except defibrinated blood, to distilled/deionized water and bring volume to 950.0mL Mix thoroughly Gently heat and bring to boiling Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C Cool to 50°– 55°C Aseptically add 50.0mL of defibrinated blood Mix thoroughly Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation of nonsporulating anaerobes.For the cultivation and maintenance of anaerobic bacteria For standardized antimicrobic susceptibility testing to determine the minimum inhibitory concentra-tions of antimicrobics for anaerobic bacteria

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Wilson Blair Base 1907

Wilkins-Chalgren Anaerobic HiVeg Agar Base

with Blood and Nonspore Anaerobic Supplement

Agar 10.0g

Plant hydrolysate 10.0g

Plant peptone 10.0g

NaCl 5.0g

Yeast extract 5.0g

L-Arginine 1.0g

Glucose 1.0g

Fe4(P2O7)3·H2O 5.0mg

Menadione 0.5mg

Nonspore anaerobic supplement

pH 7.1 ± 0.2 at 25°C

Source: This medium, without blood or nonspore anaerobic

supple-ment, is available as a premixed powder from HiMedia

Nonspore Anaerobic Supplement:

Nalidixic acid 10.0mg

Ferric pyrophosphate, soluble 5.0mg

Menadione 0.5mg

Preparation of Nonspore Anaerobic Supplement: Add

com-ponents to distilled/deionized water and bring volume to 10.0mL Mix

thoroughly Filter sterilize

blood and nonspore anaerobic supplement, to distilled/deionized water

and bring volume to 950.0mL Mix thoroughly Gently heat and bring

psi pressure–121°C Cool to 50°–55°C Aseptically add 50.0mL of

de-fibrinated blood and 10.0mL nonspore anaerobic supplement Mix

thoroughly Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation of nonsporulating anaerobes.For the cultivation

and maintenance of anaerobic bacteria For standardized antimicrobic

susceptibility testing to determine the minimum inhibitory

concentra-tions of antimicrobics for anaerobic bacteria

Wilkins-Chalgren Anaerobic HiVeg Broth Base

with Blood Compositionper liter:

Plant peptone 10.0g

Yeast extract 5.0g

NaCl 5.0g

Glucose 1.0g

L-Arginine 1.0g

Fe4(P2O7)3·H2O 5.0mg

Menadione 0.5mg

pH 7.1 ± 0.2 at 25°C

Source: This medium, without blood, is available as a premixed

pow-der from HiMedia

blood, to distilled/deionized water and bring volume to 950.0mL Mix

thoroughly Gently heat and bring to boiling Distribute into tubes or

flasks Autoclave for 15 min at 15 psi pressure–121°C Cool to 50°– 55°C Aseptically add 50.0mL of defibrinated blood Mix thoroughly Aseptically distribute into tubes or leave in flasks

Use: For the cultivation of nonsporulating anaerobes.For the cultivation and maintenance of anaerobic bacteria For standardized antimicrobic susceptibility testing to determine the minimum inhibitory concentra-tions of antimicrobics for anaerobic bacteria

Wilson and Blair’s Medium Compositionper 165.0mL:

Nutrient agar solution 100.0mL Solution B 45.0mL Solution A 20.0mL

pH 6.8 ± 0.2 at 25°C

Nutrient Agar Solution:

Agar 1.95g Pancreatic digest of gelatin 0.65g Beef extract 0.39g

Preparation of Nutrient Agar Solution: Aseptically add compo-nents to distilled/deionized water and bring volume to 100.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C

Solution A:

Na2HPO4 100.0g

Na2SO3 100.0g Glucose 50.0g Bismuth ammonium citrate 30.0g

Preparation of Solution A: Aseptically add 30.0g of bismuth am-monium citrate to 250.0mL of boiling distilled/deionized water Asep-tically add Na2SO3 to 500.0mL of boiling distilled/deionized water Mix the two solutions Add the Na2HPO4 to the boiling mixture Cool

to 45°C In a separate flask, aseptically add glucose to 250.0mL of ster-ile distilled/deionized water Mix thoroughly Aseptically add the glu-cose solution to the other cooled solution

Solution B:

Ferric citrate 2.0g Brilliant Green 0.55g

Preparation of Solution B : Aseptically add 2.0g of ferric citrate to

200.0mL of sterile distilled/deionized water Mix thoroughly Asepti-cally add Brilliant Green to 25.0mL of sterile distilled/deionized water Mix thoroughly Aseptically combine the two solutions

Preparation of Medium: Aseptically combine 100.0mL of nutrient agar solution, 20.0mL of solution A, and 45.0mL of solution B Mix thoroughly Pour into sterile Petri dishes or distribute into sterile tubes

Use: For the cultivation of Clostridium welchii.

Wilson Blair Base Composition per liter:

Agar 30.0g Proteose peptone No 3 10.0g Glucose 10.0g Beef extract 5.0g NaCl 5.0g

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1908 Wilson Blair HiVeg Agar with Brilliant Green and Selective Reagent

Selective reagent 70.0mL

Brilliant Green (1% solution) 4.0mL

pH 7.3 ± 0.2 at 25°C

Selective Reagent:

Composition per 320.2mL:

Solution 1 100.0mL

Solution 2 100.0mL

Solution 3 100.0mL

Solution 4 20.2mL

Preparation of Selective Reagent: Combine 100.0mL of solution

1, 100.0mL of solution 2, 100.0mL of solution 3, and 20.2mL of

solu-tion 4 Mix thoroughly Gently heat to boiling until a slate-grey color

develops Cool to 50°C

Solution 1:

NaHSO3 40.0g

Preparation of Solution 1: Add NaHSO3 to 100.0mL of distilled/

deionized water Mix thoroughly

Solution 2:

NaH2PO4 21.0g

Preparation of Solution 2: Add NaH2PO4 to 100.0mL of distilled/

Solution 3:

Bismuth ammonium citrate 12.5g

Preparation of Solution 3: Add bismuth ammonium citrate to

100.0mL of distilled/deionized water Mix thoroughly

Solution 4:

FeSO4 0.96g

Preparation of Solution 4: Add FeSO4 to 20.0mL of

distilled/de-ionized water Add 0.2mL of concentrated HCl Mix thoroughly

Preparation of Medium: Add components, except selective

re-agent and Brilliant Green solution, to distilled/deionized water and

bring volume to 976.0mL Mix thoroughly Gently heat and bring to

boiling Distribute into tubes or flasks Autoclave for 15 min at 15 psi

pressure–121°C Cool to 50°C Aseptically add selective reagent and

Brilliant Green solution Mix thoroughly Pour into sterile Petri dishes

or leave in tubes

Use: For the isolation and cultivation of Salmonella, especially

Salmo-nella typhi.

Wilson Blair HiVeg Agar with Brilliant Green and Selective Reagent

Agar 20.0g

Plant peptone 10.0g

Bismuth sulfite indicator 8.0g

Glucose 5.0g

Plant extract 5.0g

Na2HPO4 4.0g

FeSO4 0.3g

Brilliant Green 0.025g

Selective reagent 70.0mL

pH 7.3 ± 0.2 at 25°C

Source: This medium, without selective reagent, is available as a pre-mixed powder from HiMedia

Solution 1 100.0mL Solution 2 100.0mL Solution 3 100.0mL Solution 4 20.2mL

1, 100.0mL of solution 2, 100.0mL of solution 3, and 20.2mL of solu-tion 4 Mix thoroughly Gently heat to boiling until a slate-grey color develops Cool to 50°C

Solution 1:

NaHSO3 40.0g

Preparation of Solution 1: Add NaHSO3 to 100.0mL of distilled/ deionized water Mix thoroughly

Solution 2:

NaH2PO4 21.0g

Preparation of Solution 2: Add NaH2PO4 to 100.0mL of distilled/ deionized water Mix thoroughly

Solution 3:

Preparation of Solution 3: Add bismuth ammonium citrate to 100.0mL of distilled/deionized water Mix thoroughly

Solution 4:

FeSO4 0.96g

Preparation of Solution 4: Add FeSO4 to 20.0mL of distilled/de-ionized water Add 0.2mL of concentrated HCl Mix thoroughly

Preparation of Medium: Add components, except selective re-agent and Brilliant Green solution, to distilled/deionized water and bring volume to 976.0mL Mix thoroughly Gently heat and bring to boiling Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C Cool to 50°C Aseptically add selective reagent and Brilliant Green solution Mix thoroughly Pour into sterile Petri dishes

or leave in tubes

Use: For the isolation and cultivation of Salmonella, especially Salmo-nella typhi.

Wilson Blair HiVeg Agar Base with Selective Reagent and Brilliant Green Compositionper liter:

Agar 30.0g Glucose 10.0g Plant special peptone 10.0g Plant extract 5.0g NaCl 5.0g Selective reagent 70.0mL Brilliant Green (1% solution) 4.0mL

pH 7.3 ± 0.2 at 25°C

Source: This medium, without selective reagent or Brilliant Green, is available as a premixed powder from HiMedia

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Winogradsky’s N-Free Medium 1909

Solution 1 100.0mL

Solution 2 100.0mL

Solution 3 100.0mL

Solution 4 20.2mL

1, 100.0mL of solution 2, 100.0mL of solution 3, and 20.2mL of

solu-tion 4 Mix thoroughly Gently heat to boiling until a slate-grey color

develops Cool to 50°C

Solution 1:

NaHSO3 40.0g

Preparation of Solution 1: Add NaHSO3 to 100.0mL of distilled/

Solution 2:

NaH2PO4 21.0g

Preparation of Solution 2: Add NaH2PO4 to 100.0mL of distilled/

Solution 3:

Preparation of Solution 3: Add bismuth ammonium citrate to

100.0mL of distilled/deionized water Mix thoroughly

Solution 4:

FeSO4 0.96g

Preparation of Solution 4: Add FeSO4 to 20.0mL of

distilled/de-ionized water Add 0.2mL of concentrated HCl Mix thoroughly

Preparation of Medium: Add components, except selective

re-agent and Brilliant Green solution, to distilled/deionized water and

bring volume to 976.0mL Mix thoroughly Gently heat and bring to

boiling Distribute into tubes or flasks Autoclave for 15 min at 15 psi

pressure–121°C Cool to 50°C Aseptically add selective reagent and

Brilliant Green solution Mix thoroughly Pour into sterile Petri dishes

or leave in tubes

Use: For the isolation and cultivation of Salmonella, especially

Salmo-nella typhi.

Winge Agar Compositionper liter:

Glucose 20.0g

Agar 15.0g

Yeast extract 3.0g

pH 7.2 ± 0.2 at 25°C

water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.2

Gently heat and bring to boiling Distribute into tubes or flasks

Auto-clave for 15 min at 15 psi pressure–121°C Pour into sterile Petri dishes

or leave in tubes

Use: For the cultivation and maintenance of Candida albicans,

Can-dida kefyr, CanCan-dida tropicalis, Citeromyces matritensis, Cryptococcus

albidus, Neurospora crassa, Octosporomyces octosporus, Neurospora

sitophila, and Saccharomyces species.

Winge Melibiose Agar Compositionper liter:

Agar 15.0g Melibiose 10.0g Yeast extract 3.0g

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.2 Gently heat and bring to boiling Distribute into tubes or flasks Auto-clave for 15 min at 15 psi pressure–121°C Pour into sterile Petri dishes

or leave in tubes

Use: For the cultivation and maintenance of Saccharomyces

cerevi-siae.

Winogradsky’s Medium, Modified Compositionper liter:

CaCO3 5.0g (NH4)2SO4 1.0g

K2HPO4 1.0g NaCl 1.0g MgSO4·7H2O 0.5g FeSO4 0.4g

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Gently heat until dis-solved Do not autoclave Distribute into tubes or flasks Swirl flask while dispensing to suspend precipitate

Use: For the cultivation of nitrifying bacteria

Winogradsky’s N-Free Medium Compositionper liter:

Agar 20.0g CaCO3 5.0mg Sugar solution 100.0mL Concentrated salt solution 5.0mL

pH 7.2 ± 0.2 at 25°C

Sugar Solution:

Sucrose or glucose 10.0g

Preparation of Sugar Solution: Add sucrose or glucose to 100.0mL

of distilled/deionized water Mix thoroughly Autoclave for 10 min at 10 psi pressure–115°C Cool to 50°C

Concentrated Salt Solution:

KH2PO4 50.0g MgSO4·7H2O 25.0g NaCl 25.0g FeSO4·7H2O 1.0g MnSO4·4H2O 1.0g

Na2MoO4·4H2O 1.0g

Preparation of Concentrated Salt Solution: Add components

to tap water and bring volume to 1.0L Mix thoroughly Filter sterilize

Preparation of Medium: Add components, except sugar solution,

to distilled/deionized water and bring volume to 900.0mL Mix thor-oughly Gently heat and bring to boiling Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C Cool to 50°C Asepti-cally add sugar solution Adjust pH to 7.2 Mix thoroughly Pour into sterile Petri dishes or leave in tubes

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1910 Winogradsky’s Nitrite Medium

Use: For the cultivation and maintenance of Azomonas insignis.

Winogradsky’s Nitrite Medium

Agar 15.0g

NaNO2 2.0g

Na2CO3 , anhydrous 1.0g

K2HPO4 0.5g

to boiling Distribute into tubes Autoclave for 15 min at 15 psi

pres-sure–121°C

Use: For the selective isolation and cultivation of Nocardia species

and Rhodococcus species.

WL Differential Agar (Wallerstein Laboratory Differential Agar)

Composition per liter:

Glucose 50.0g

Agar 20.0g

Yeast extract 4.0g

KH2PO4 0.55g

KCl 0.425g

CaCl2·2H2O 0.125g

MgSO4·7H2O 0.125g

Bromcresol Green 0.022g

Actidione® (cycloheximide) 4.0mg

FeCl3 2.5mg

MnSO4·4H2O 2.5mg

pH 5.5 ± 0.2 at 25°C

Caution: Cycloheximide is toxic Avoid skin contact or aerosol

for-mation and inhalation

Di-agnostic Systems

water and bring volume to 1.0L Mix thoroughly Gently heat with

mix-ing and brmix-ing to boilmix-ing Distribute into tubes or flasks Autoclave for

15 min at 15 psi pressure–121°C Pour into sterile Petri dishes or leave

in tubes

Use: For the differential cultivation of bacteria from industrial

fermen-tation processes Growth of yeasts and molds is inhibited

WL Differential HiVeg Agar

Glucose 50.0g

Agar 20.0g

Yeast extract 4.0g

KH2PO4 0.55g

KCl 0.425g

MgSO4 0.125g

CaCl2 0.125g

Cycloheximide 4.0mg

FeCl3 2.5mg

MnSO4 2.5mg

pH 5.5 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Hi-Media

Caution: Cycloheximide is toxic Avoid skin contact or aerosol for-mation and inhalation

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Gently heat with mix-ing and brmix-ing to boilmix-ing Distribute into tubes or flasks Autoclave for

in tubes

Use: For the differential cultivation of bacteria from industrial fermen-tation processes Growth of yeasts and molds is inhibited For the selective isolation and enumeration of bacteria encountered in brewer-ies and industrial fermentations

WL Differential HiVeg Broth Compositionper liter:

Glucose 50.0g Plant hydrolysate 5.0g Yeast extract 4.0g

KH2PO4 0.55g KCl 0.425g MgSO4 0.125g CaCl2 0.125g Bromcresol Green 0.022g Cycloheximide 4.0mg FeCl3 2.5mg MnSO4 2.5mg

pH 5.5 ± 0.2 at 25°C

Caution: Cycloheximide is toxic Avoid skin contact or aerosol for-mation and inhalation

15 min at 15 psi pressure–121°C

WL Differential Medium (Wallerstein Laboratory Differential Medium) Compositionper liter:

Glucose 50.0g Agar 20.0g Pancreatic digest of casein 5.0g Yeast extract 4.0g

KH2PO4 0.55g KCl 0.425g CaCl2·2H2O 0.125g MgSO4·7H2O 0.125g Bromcresol Green 0.022g Actidione® (cycloheximide) 4.0mg FeCl3 2.5mg MnSO4·4H2O 2.5mg

pH 5.5 ± 0.2 at 25°C

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WL Nutrient Agar 1911

Di-agnostic Systems

in tubes

WL Differential Medium (Wallerstein Laboratory Differential Medium)

Glucose 50.0g

Agar 20.0g

Yeast extract 4.0g

KH2PO4 0.55g

KCl 0.425g

CaCl2·2H2O 0.125g

MgSO4·7H2O 0.125g

Actidione® (cycloheximide) 4.0mg

FeCl3 2.5mg

MnSO4·4H2O 2.5mg

Na2CO3 (1% solution) 30.0mL

pH 6.5 ± 0.2 at 25°C

Di-agnostic Systems

in tubes

WL Medium with Tomato Juice

(Wallerstein Laboratory Medium with Tomato Juice)

Glucose 50.0g

Yeast extract 4.0g

KH2PO4 0.55g

KCl 0.425g

CaCl2 0.125g

MgSO4·7H2O 0.125g

FeCl3 2.5mg

MnSO4·4H2O 2.5mg

Tomato juice, canned, clarified 400.0mL

pH 5.5 ± 0.2 at 25°C

Use: For the cultivation of microorganisms from alcoholic mash

WL Nutrient Agar (Wallerstein Laboratory Nutrient Agar) Compositionper liter:

KH2PO4 0.55g KCl 0.425g CaCl2·2H2O 0.125g MgSO4·7H2O 0.125g Bromcresol Green 0.022g FeCl3 2.5mg MnSO4·4H2O 2.5mg

pH 5.5 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from BD Di-agnostic Systems and Oxoid Unipath

in tubes

Use: For the detection of bacteria and yeasts in industrial fermentation processes, particularly from beer processing

WL Nutrient Agar (Wallerstein Laboratory Nutrient Agar) Compositionper liter:

KH2PO4 0.55g KCl 0.425g CaCl2·2H2O 0.125g MgSO4·7H2O 0.125g Bromcresol Green 0.022g FeCl3 2.5mg MnSO4·4H2O 2.5mg

pH 6.5 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from BD Di-agnostic Systemsand Oxoid Unipath

in tubes

Use: For the detection of bacteria and yeasts from industrial fermenta-tion processes, particularly from beer processing

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1912 WL Nutrient Broth

WL Nutrient Broth (Wallerstein Laboratory Nutrient Broth)

Glucose 50.0g

Yeast extract 4.0g

KH2PO4 0.55g

KCl 0.425g

CaCl2 0.125g

MgSO4·7H2O 0.125g

FeCl3 2.5mg

MnSO4·4H2O 2.5mg

pH 5.5 ± 0.2 at 25°C

Di-agnostic Systems and Oxoid Unipath

water and bring volume to 1.0L Mix thoroughly Distribute into tubes

Use: For the cultivation of yeasts, molds, and bacteria found in

brew-ing and other industrial fermentation processes

Glucose 50.0g

Yeast extract 4.0g

KH2PO4 0.55g

KCl 0.425g

CaCl2 0.125g

MgSO4·7H2O 0.125g

FeCl3 2.5mg

MnSO4·4H2O 2.5mg

pH 6.5 ± 0.2 at 25°C

water and bring volume to 1.0L Mix thoroughly Distribute into tubes

Use: For the cultivation of yeasts, molds, and bacteria found in

brew-ing and other industrial fermentation processes

Glucose 50.0g

Yeast extract 4.0g

FeCl3 2.5g

MnSO4·4H2O 2.5g

KH2PO4 0.55g

KCl 0.425g

CaCl2·2H2O 0.125g

MgSO4·7H2O 0.125g

pH 5.5 ± 0.2 at 25°C

Use: For the control of brewing and other fermentation processes

WL Nutrient HiVeg Broth Compositionper liter:

Glucose 50.0g Plant hydrolysate 5.0g Yeast extract 4.0g

KH2PO4 0.55g KCl 0.425g MgSO4 0.125g CaCl2 0.125g Bromcresol green 0.022g FeCl3 2.5mg MnSO4 2.5mg

pH 5.5 ± 0.2 at 25°C

WL Nutrient HiVeg Medium Compositionper liter:

Glucose 50.0g Agar 20.0g Plant hydrolysate 5.0g Yeast extract 4.0g

KH2PO4 0.55g KCl 0.425g CaCl2 0.125g MgSO4 0.125g Bromcresol green 0.022g FeCl3 2.5mg MnSO4 2.5mg

pH 5.5 ± 0.2 at 25°C

WMC Medium Compositionper 1010.0mL:

NaHCO3 5.0g Sodium acetate 1.0g

L-Cysteine 0.5g

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Wolinella succinogenes Medium 1913

Resazurin 1.0mg

Mineral salt solution 2xW 500.0mL

LIP solution 50.0mL

TYC solution 50.0mL

Trace elements solution 10.0mL

Na2S·9H2O solution 10.0mL

pH 7.2 ± 0.2 at 25°C

Mineral Solution 2xW:

NaCl 40.0g

MgCl2·6H2O 5.6g

MgSO4·7H2O 0.7g

KCl 0.68g

NH4Cl 0.5g

CaCl2·2H2O 0.28g

K2HPO4 0.28g

Preparation of Mineral Solution 2xW: Add components to

dis-tilled/deionized water and bring volume to 1.0L Mix thoroughly Filter

sterilize Store at room temperature in the dark

LIP Solution:

L-Isoleucine 10.0g

L-Leucine 5.0g

Pantothenate 0.1g

Preparation of LIP Solution : Add components to

distilled/deion-ized water and bring volume to 1.0L Mix thoroughly May be stored

unsterilized at −20°C

TYC Solution:

Casamino acids 100.0g

Yeast extract 50.0g

L-Tryptophan 1.0g

Preparation of TYC Solution : Add components to

distilled/de-ionized water and bring volume to 1.0L Mix thoroughly Sparge with

100% N2 Autoclave for 15 min at 15 psi pressure–121°C

Trace Elements Solution:

MgSO4·7H2O 3.0g

Nitrilotriacetic acid 1.5g

NaCl 1.0g

MnSO4·2H2O 0.5g

CoSO4·7H2O 0.18g

ZnSO4·7H2O 0.18g

FeSO4·7H2O 0.1g

CaCl2·2H2O 0.1g

KAI(SO4)2·12H2O 0.02g

CuSO4·5H2O 0.01g

H3BO3 0.01g

Na2MoO4·2H2O 0.01g

NiCl2·6H2O 0.025g

Na2SeO3·5H2O 0.3mg

Preparation of Trace Elements Solution: Add nitrilotriacetic

acid to 500.0mL of distilled/deionized water Adjust pH to 6.5 with

KOH Add remaining components Add distilled/deionized water to

1.0L

Na 2 S·9H 2 O Solution:

Na2S·9H2O 0.5g

Preparation of Na 2 S·9H 2 O Solution: Add Na2S·9H2O to dis-tilled/deionized water and bring volume to 10.0mL Mix thoroughly Sparge with 100% N2 Autoclave for 15 min at 15 psi pressure–121°C

Preparation of Medium: Prepare and dispense medium under 80%

H2 + 20% CO2 Add components, except TYC solution and Na2S·9H2O solution, to distilled/deionized water and bring volume to 940.0mL Mix thoroughly Sparge with 80% H2 + 20% CO2 Autoclave for 15 min at 15 psi pressure–121°C Aseptically and anaerobically add 50.0mL of sterile TYC solution and 10.0mL of sterile Na2S·9H2O so-lution Mix thoroughly

Use: For the cultivation and maintenance of Methanococcus voltae.

Wolin-Bevis Medium Compositionper liter:

Agar 20.0g (NH4)2SO4 1.0g

KH2HPO4 1.0g Glucose 0.25g

L-Histidine·HCl 0.25g Tween™ 80 (polysorbate 80) 3.0mL

pH 5.4 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Gently heat and bring

to boiling Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C Pour into sterile Petri dishes or leave in tubes

Use: For the enhanced production of chlamydospores by Candida albicans.

Wolinella succinogenes Medium

K2HPO4 5.0g Fumaric acid 3.0g Sodium formate 3.0g (NH4)2SO4 1.0g Yeast extract 1.0g MgCl2·6H2O 0.2g FeSO4·7H2O 0.02g Resazurin 1.0mg Sodium thioglycolate solution 10.0mL

pH 7.0 ± 0.2 at 25°C

Sodium Thioglycolate Solution:

Sodium thioglycolate 0.5g

Preparation of Sodium Thioglycolate Solution: Add sodium thioglycolate to distilled/deionized water and bring volume to 10.0mL Mix thoroughly Sparge with 100% N2 Autoclave for 15 min at 15 psi pressure–121°C

Preparation of Medium: Add components, except sodium thiogly-colate solution, to distilled/deionized water and bring volume to 990.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure– 121°C Aseptically add 10.0mL of sterile sodium thioglycolate solu-tion Mix thoroughly Aseptically distribute into sterile tubes or flasks

Use: For the cultivation and maintenance of Wolinella succinogenes.

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1914 Woods and Welton Agar

Woods and Welton Agar Compositionper liter:

NaCl 23.4g

Casein hydrolysate 17.0g

Agar 15.0g

Glycerol 10.0g

Pancreatic digest of gelatin 5.0g

Glucose 5.0g

Papaic digest of soybean meal 3.0g

Beef extract 3.0g

Yeast extract 2.0g

Casamino acids, vitamin free 0.5g

Na2SO3 0.1g

pH 7.6 ± 0.2 at 25°C

to boiling Adjust pH to 7.6 Distribute into tubes or flasks Autoclave

for 15 min at 15 psi pressure–121°C Pour into sterile Petri dishes or

leave in tubes

Use: For the cultivation of Vibrio alginolyticus.

Worfel-Ferguson Agar Compositionper liter:

Sucrose 20.0g

Agar 15.0g

NaCl 2.0g

Yeast extract 2.0g

K2SO4 1.0g

MgSO4·7H2O 0.25g

pH 6.5 ± 0.2 at 25°C

psi pressure–121°C Pour into sterile Petri dishes or leave in tubes

Use: For the detection of capsule production by Klebsiella For

sero-logical detection of the Neufeld (Quellung) reaction

Wort Agar Compositionper liter:

Agar 15.0g

Malt extract 15.0g

Maltose 12.75g

Dextrin 2.75g

Glycerol 2.35g

K2HPO4 1.0g

NH4Cl 1.0g

Pancreatic digest of gelatin 0.78g

pH 4.8 ± 0.2 at 25°C

to boiling Boil for 1 min with mixing Distribute into tubes or flasks

Au-toclave for 15 min at 15 psi pressure–121°C Do not overheat, as this will

result in hydrolysis of the agar An additional 5.0g of agar can be used to

make a firmer agar Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation and enumeration of yeasts The low pH of the agar selectively inhibits bacterial growth

Wort Agar Compositionper liter:

Agar 25.0g Yeast extract 1.0g Wort solution 1.0L

Wort Solution:

Malt extract 110.0g

Preparation of Wort Solution : Add malt extract to

distilled/deion-ized water and bring volume to 1.0L Mix thoroughly

Preparation of Medium: Combine components Mix thoroughly Gently heat and bring to boiling Distribute into tubes or flasks Auto-clave for 15 min at 15 psi pressure–121°C Pour into sterile Petri dishes

or leave in tubes

Use: For the cultivation and mainteneance of a wide variety of yeasts

and filamentous fungi

Wort Agar 6°Brix Compositionper liter:

Malt 250.0g Agar 20.0g

Preparation of Medium: Add 250.0g of ground malt to tap water and bring volume to 1.0L Mix thoroughly Gently heat to 55°–60°C Maintain at 55°–60°C for 1.5–2.0 hr while mixing regularly Elevate temperature to 80°C and maintain for 10 min with thorough mixing Cool the wort to 25°C and filter through a linen bag Adjust the con-centration of sugars to 6° Brix (specific gravity of 1.024 at 24°C) Add 20.0g of agar Gently heat and bring to boiling Mix thoroughly Dis-tribute into tubes or flasks Autoclave for 30 min at 3 psi pressure– 105°C Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation and maintenance of Rhodococcus erythropo-lis.

Wort Broth Compositionper liter:

Yeast extract 1.0g Wort solution 1.0L

Wort Solution:

Malt extract 110.0g

Preparation of Wort Solution : Add malt extract to

distilled/deion-ized water and bring volume to 1000.0mL Mix thoroughly

Preparation of Medium: Combine components Mix thoroughly Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure– 121°C

Use: For the cultivation of a wide variety of yeasts and filamentous fungi

Wort HiVeg Agar Compositionper liter:

Agar 15.0g Malt extract 15.0g Maltose 12.75g Dextrin 2.75g

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