Handbook of Microbiological Media, Fourth Edition part 92 potx

0.4g Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L.. Preparation of Medium: Add components to distilled/deionized water and bring volume to

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Kohn Two Tube HiVeg Medium No 1 Base with Urea 905

sterile Petri dishes Allow plates to dry in the dark at 30°C for 24 hr

be-fore using

Use: For the isolation and cultivation of acidogenic microorganisms

from foods For the differentiation of citrate-fermenting lactic bacteria,

such as Lactobacillus lactis subspecies diacetylactis, from the

nonci-trate-fermenting Lactobacillus lactis subspecies cremoris.

Knisely Medium for Bacillus anthracis

Compositionper liter:

Beef heart, solids from infusion 500.0g

Agar 15.0g

Pancreatic digest of casein 10.0g

NaCl 5.0g

EDTA 200.0mg

Lysozyme 40.0mg

Thallous acetate 40.0mg

Polymyxin 30,000U

Preparation of Medium: Add components, except EDTA,

lysozyme, thallous acetate, and polymyxin, to distilled/deionized water

and bring volume to 1.0mL Mix thoroughly Gently heat and bring to

boiling Adjust pH to 7.3 Autoclave for 15 min at 15 psi pressure–

121°C Cool to 45°–50°C Aseptically add sterile EDTA, lysozyme,

thallous acetate, and polymyxin Mix thoroughly Pour into sterile Petri

dishes or distribute into sterile tubes

Use: For the cultivation and maintenance of Bacillus anthracis.

Koch’s K1 Medium

Glucose 1.8g

Peptone 0.6g

Yeast extract 0.4g

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 of a variety of fungi

Kohn Two Tube Medium No 1 Base with Urea

Agar 16.0g

Peptic digest of animal tissue 15.0g

Mannitol 10.0g

Beef extract 2.0g

Yeast extract 2.0g

Glucose 1.0g

Phenol Red 0.05g

Urea solution 25.0mL

pH 7.3 ± 0.2 at 25°C

Source: This medium, without urea solution, is available as a

pre-mixed powder from HiMedia

Urea Solution:

Compositionper 100.0mL:

Urea 5.0g

Preparation of Urea Solution: Add urea to distilled/deionized

wa-ter and bring volume to 100.0mL Mix thoroughly Filwa-ter swa-terilize

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

to boiling Distribute into tubes Autoclave for 15 min at 10 psi pressure– 115°C Cool to 60°C Aseptically add 25mL of sterile urea solution Mix thoroughly Allow to solidify as a slant in tubes with a genrous butt

Use: For the identification of Enterobacteriaceae on the basis of glu-cose and mannitol fermentation and urease production

Kohn Two Tube Medium No 2

Casein enzymic hydrolysate 10.0g Peptic digest of animal tissue 10.0g Salicin 10.0g Sucrose 10.0g NaCl 5.0g Agar 3.0g

Na2HPO4 0.09g Bromthymol Blue 0.02g

Na2S2O3 0.016g

pH 7.3 ± 0.2 at 25°C

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

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 Autoclave for 15 min at 10 psi pressure– 115°C Allow to solidify in tubes in an upright position

Use: For the identification of members of Enterobacteriaceae on the basis of sucrose and salicin fermentation, motility, H2S production, and indole production

Kohn Two Tube HiVeg Medium No 1 Base with Urea

Agar 16.0g Plant peptone 15.0g Mannitol 10.0g Plant extract 2.0g Yeast extract 2.0g Glucose 1.0g Phenol Red 0.05g Urea solution 25.0mL

pH 7.3 ± 0.2 at 25°C

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

Urea Solution:

Urea 5.0g

Preparation of Urea Solution: Add urea to distilled/deionized wa-ter and bring volume to 100.0mL Mix thoroughly Filwa-ter swa-terilize

to boiling Distribute into tubes Autoclave for 15 min at 10 psi pressure– 115°C Cool to 60°C Aseptically add 25.0mL of sterile urea solution Mix htoroughly Allow to solidify as a slant in tubes with a genrous butt

Use: For the identification of Enterobacteriaceae on the basis of glu-cose and mannitol fermentation and urease production

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906 Kohn Two Tube HiVeg Medium No 2

Kohn Two Tube HiVeg Medium No 2

Plant hydrolysate 10.0g

Plant peptone 10.0g

Salicin 10.0g

Sucrose 10.0g

NaCl 5.0g

Agar 3.0g

Na2HPO4 0.09g

Bromthymol Blue 0.02g

Na2S2O3 0.016g

pH 7.3 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from

Hi-Media

to boiling Distribute into tubes Autoclave for 15 min at 10 psi pressure–

115°C Allow to solidify in tubes in an upright position

Use: For the identification of members of Enterobacteriaceae on the

basis of sucrose and salicin fermentation, motility, H2S production, and

indole production

KoKo Medium

K2HPO4 1.6g

NaH2PO4·2H2O 1.0g

Peptone, meat 1.0g

Yeast extract 1.0g

NH4Cl 0.5g

MgSO4·6H2O 0.16g

Resazurin 0.5mg

Glucose solution 100.0mL

NaHCO3 solution 10.0mL

CaCl2·2H2O solution 10.0mL

L-Cysteine·HCl·H2O solution 10.0mL

Na2S·9H2O solution 10.0mL

Trace elements solution SL-4 10.0mL

Wolfe’s vitamin solution 10.0mL

pH 7.0 ± 0.2 at 25°C

Glucose Solution:

D-Glucose 5.0g

Preparation of Glucose Solution: Add glucose to

distilled/deion-ized water and bring volume to 100.0mL Mix thoroughly Sparge with

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

NaHCO 3 Solution:

NaHCO3 1.0g

Preparation of NaHCO 3 Solution: Add NaHCO3 to

distilled/de-ionized water and bring volume to 10.0mL Mix thoroughly Sparge

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

CaCl 2 Solution:

CaCl2·2H2O 0.06g

Preparation of CaCl 2 Solution: Add CaCl2·2H2O to

distilled/de-ionized water and bring volume to 10.0mL Mix thoroughly Sparge

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

L -Cysteine·HCl·H 2 O Solution:

L-Cysteine·HCl·H2O 0.3g

Preparation of L -Cysteine·HCl·H 2 O Solution: Add L -cysteine·HCl·H2O 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

Na 2 S·9H 2 O Solution:

Na2S·9H2O 0.3g

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 Before use, neutralize to pH 7.0 with sterile HCl

Trace Elements Solution SL-4:

EDTA 0.5g FeSO4·7H2O 0.2g Trace elements solution SL-6 100.0mL

Trace Elements Solution SL-6:

MnCl2·4H2O 0.5g

H3BO3 0.3g CoCl2·6H2O 0.2g ZnSO4·7H2O 0.1g

Na2MoO4·2H2O 0.03g NiCl2·6H2O 0.02g CuCl2·2H2O 0.01g

Preparation of Trace Elements Solution SL-6 : Add components

to distilled/deionized water and bring volume to 1.0L Mix thoroughly

Preparation of Trace Elements Solution SL-4: Add compo-nents to distilled/deionized water and bring volume to 1.0L Mix thor-oughly Filter sterilize

Wolfe’s Vitamin Solution:

Pyridoxine·HCl 10.0mg

p-Aminobenzoic acid 5.0mg

Lipoic acid 5.0mg Nicotinic acid 5.0mg Riboflavin 5.0mg Thiamine·HCl 5.0mg Calcium DL-pantothenate 5.0mg Biotin 2.0mg Folic acid 2.0mg Vitamin B12 0.1mg

Preparation of Wolfe’s Vitamin Solution: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Sparge with 100% N2 Filter sterilize

Preparation of Medium: Prepare and dispense medium under 100% N2 Add components, except glucose solution, NaHCO3 solu-tion, CaCl2·2H2O solution, L-cysteine·HCl·H2O solution, Na2S·9H2O solution, and Wolfe’s vitamin solution, to distilled/deionized water and bring volume to 850.0mL Mix thoroughly Adjust pH to 7.0 Sparge with 100% N2 Autoclave for 15 min at 15 psi pressure–121°C Asep-tically and anaerobically add 100.0mL of sterile glucose solution, 10.0mL of sterile NaHCO3 solution, 10.0mL of sterile CaCl2·2H2O so-lution, 10.0mL of sterile L-cysteine·HCl·H2O solution, 10.0mL of ster-ile Na2S·9H2O solution, and 10.0mL of sterile Wolfe’s vitamin

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KPL Medium 907

solution Mix thoroughly The pH should be 7.0 A buffer solution of

1% MOPS from a 10% anaerobic solution at pH 6.9–7.0 may be added

aseptically and anaerobically to enhance the buffer capacity of the

me-dium Aseptically and anaerobically distribute into sterile tubes or

bot-tles

Use: For the cultivation of Thermoanaerobacter italicus.

Korthof Medium

NaCl 1.4g

Na2HPO4·2H2O 0.88g

Peptone 0.8g

KH2PO4 0.24g

CaCl2 0.04g

KCl 0.04g

NaHCO3 0.02g

Rabbit serum, inactivated 80.0mL

Rabbit hemoglobin solution 8.0mL

pH 7.2 ± 0.2 at 25°C

Rabbit Hemoglobin Solution:

Rabbit blood clot 10.0mL

Preparation of Rabbit Hemoglobin Solution: Add rabbit blood

clot to 10.0mL of distilled/deionized water Lyse the clot by freezing

and thawing

Preparation of Medium: Add components, except rabbit serum

and rabbit hemoglobin solution, to distilled/deionized water and bring

volume to 1.0L Mix thoroughly Gently heat and bring to boiling Cool

to 25°C Filter through Whatman #1 filter paper Distribute into flasks

in 100.0mL volumes Autoclave for 15 min at 15 psi pressure–121°C

Cool to 45°–50°C Aseptically add 8.0mL of rabbit serum and 0.8mL

of rabbit hemoglobin solution to each flask Mix thoroughly

Use: For the cultivation of Leptospira species.

Korthof Medium, Modified

NaCl 1.4g

Na2HPO4·2H2O 0.88g

Peptone 0.8g

KH2PO4 0.24g

CaCl2 0.04g

KCl 0.04g

NaHCO3 0.02g

Rabbit serum, heat inactivated at 56°C 100.0mL

pH 7.2–7.6 at 25°C

Preparation of Medium: Add components, except rabbit serum, to

distilled/deionized water and bring volume to 900.0L Mix thoroughly

Gently heat and bring to boiling Boil for 20 min Cool overnight at

4°C Filter through Whatman #2 filter paper Distribute into tubes or

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

56°C Aseptically add 100.0mL of rabbit serum Mix thoroughly

Use: For the cultivation of Leptospira species

Koser Citrate Broth (BAM M72)

Sodium citrate 3.0g

NaNH4HPO4·4H2O 1.5g

KH2PO4 1.0g MgSO4·7H2O 0.2g

pH 6.7 ± 0.2 at 25°C

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

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 differentiation of Escherichia coli and Enterobacter

aero-genes based on citrate utilization.

Koser Citrate Medium

Sodium citrate 3.0g NaNH4HPO4·4H2O 1.5g

KH2PO4 1.0g MgSO4·7H2O 0.2g

pH 6.7 ± 0.2 at 25°C

Use: For the differentiation of Escherichia coli and Enterobacter

aero-genes based on citrate utilization.

Kosmachev’s Medium

Agar 15.0g CaCO3 4.0g KNO3 1.0g (NH4)2SO4 1.0g

Na2HPO4 1.0g MgSO4·7H2O 0.5g FeSO4·7H2O 0.01g Yeast autolysate (30% solution) 15.0mL

Use: For the isolation and cultivation of Actinomadura species,

Acti-nopolyspora species, Excellospora species, and Microspora species.

KPL Medium

Agar 15.0g Galactose 10.0g Glucose 10.0g Lactic acid whey 1.0L White table wine 140.0mL Tween™ 80 1.0mL

pH 5.5 ± 0.2 at 25°C

Lactic Acid Whey:

Skim milk (10% solution) 1.0L

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908 Kracke Blood Culture HiVeg Medium

Preparation of Lactic Acid Whey: Adjust the pH of the skim

milk to 5.5 with lactic acid Gently heat and bring to boiling Continue

boiling for 30 min Filter through Whatman #1 filter paper

Preparation of Medium: Combine components, except white table

wine Mix thoroughly Gently heat and bring to boiling Adjust pH to

5.5 Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C

Filter sterilize white table wine To cooled, sterile basal medium,

asep-tically add sterile white table wine Mix thoroughly Pour into sterile

Petri dishes or distribute into sterile tubes

Use: For the cultivation and maintenance of Lactobacillus

kefiranofa-ciens.

Kracke Blood Culture HiVeg Medium

NaCl 49.0g

Glucose 10.0g

Plant peptone No 3 10.0g

Na2HPO4 2.0g

Plant infusion 2.0g

Plant special infusion 1.0g

Sodium citrate 1.0g

pH 7.4 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from

Hi-Media

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

psi pressure–121°C

Use: For the cultivation of pathogens in cases of bacteremia The

medium is inoculated with blood from a patient Approximately

10-15mL of blood normally is inoculated into 50mL of medium

Krainsky’s Asparagine Agar

Agar 15.0g

Glucose 10.0g

K2HPO4 0.5g

L-Asparagine 0.5g

pH 7.0 ± 0.2 at 25°C

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

Use: For the cultivation and maintenance of Streptomyces

fragmen-tans.

KRANEP Agar Base

KSCN 25.5g

Agar 18.3g

Sodium pyruvate 8.2g

Pancreatic digest of gelatin 6.1g

LiCl 5.1g

Mannitol 5.1g

Beef extract 3.7g

NaN3 0.05g

Cycloheximide 0.041g Egg yolk emulsion 100.0mL

pH 6.8 ± 0.2 at 25°C

Caution: Sodium azide is toxic Azides also react with metals and disposal must be highly diluted

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

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

Egg Yolk Emulsion:

Composition: Chicken egg yolks 11 Whole chicken egg 1

Preparation of Egg Yolk Emulsion: Soak eggs with 1:100 dilu-tion of saturated mercuric chloride soludilu-tion for 1 min Crack eggs and separate yolks from whites Mix egg yolks with 1 chicken egg

Preparation of Medium: Add components, except egg yolk emul-sion, 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–121°C Cool to 45°–50°C Aseptically add 100.0mL of egg yolk emulsion Mix thoroughly Pour into sterile Petri dishes or distribute into sterile tubes

Use: For the isolation and enumeration of staphylococci from foods

KRANEP HiVeg Agar Base with Egg Yolk Emulsion

Potassium thiocyanate 25.5g Agar 15.0g Sodium pyruvate 8.2g LiCl 5.1g Mannitol 5.1g Plant peptone 5.0g NaCl 5.0g Plant extract 1.5g Yeast extract 1.5g NaN3 0.05g Cycloheximide 0.041g Egg yolk emulsion 100.0mL

pH 6.8 ± 0.2 at 25°C

Source: This medium, without egg yolk emulsion, is available as a premixed powder from HiMedia

Caution: Sodium azide is toxic Azides also react with metals and disposal must be highly diluted

Caution: Lithium Chloride is harmful Avoid bodily contact and in-halation of vapors On contact with skin wash with plenty of water im-mediately

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

Egg Yolk Emulsion:

Egg yolks 30.0mL NaCl, 0.9% solution 70.0mL

Preparation of Egg Yolk Emulsion: Soak eggs with 1:100 dilu-tion of saturated mercuric chloride soludilu-tion for 1 min Crack 11 eggs and separate yolks from whites Mix egg yolks Measure 30.0mL of egg yolk emulsion and add to 70.0mL of 0.9% sterile NaCl solution Mix thoroughly Warm to 45°–50°C

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Kupferberg Trichomonas Broth 909

Preparation of Medium: Add components, except egg yolk

emul-sion, 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–121°C Cool to 45°–50°C Aseptically add 100.0mL of

egg yolk emulsion Mix thoroughly Pour into sterile Petri dishes or

distribute into sterile tubes

Use: For the isolation and enumeration of staphylococci from foods

Kreb’s Yeast Lactate Medium

Yeast extract 10.0g

Na2HPO4·2H2O 3.0g

KH2PO4 1.0g

Sodium lactate solution 40.0mL

pH 7.0 ± 0.2 at 25°C

Sodium Lactate Solution:

Sodium lactate 70.0g

Preparation of Sodium Lactate Solution: Add sodium lactate to

distilled/deionized water and bring volume to 100.0mL Mix

thorough-ly Filter sterilize

Preparation of Medium: Add components, except sodium lactate

solution, to distilled/deionized water and bring volume to 960.0mL

Mix thoroughly Bring pH to 7.0 Autoclave for 15 min at 15 psi

pres-sure–121°C Aseptically add 40.0mL of sterile sodium lactate solution

Mix thoroughly Aseptically distribute into sterile tubes or flasks

Use: For the cultivation of Propionibacterium acidipropionici,

onibacterium freudenreichii, Propionibacterium intermedium,

Propi-onibacterium jensenii, PropiPropi-onibacterium species, and

Propionibacte-rium thoenii.

Kundrant Agar

Composition per liter:

Agar 10.0g

Meat peptone 7.8g

Casein peptone 7.8g

Starch 4.0g

Gelatin 4.0g

NaCl 3.0g

Yeast extract 2.8g

Glucose 1.0g

Bromcresol Purple 1.6mg

pH 6.8 ± 0.2 at 25°C

Source: This medium is available from HiMedia

psi pressure–121°C

Use: For the qualitative detection of residues from sulfonamides and

other antimicrobics in animal-derived foods

Kunkee Medium

Glucose 5.0g

Peptone 5.0g

Yeast extract 5.0g Filtered tomato juice 250.0mL Ethanol (96% solution) 176.0mL Tween™ 80 0.5mL

pH 5.5 ± 0.2 at 25°C

Preparation of Medium: Add components, except ethanol, to dis-tilled/deionized water and bring volume to 1.0L Mix thoroughly Dis-tribute into tubes in 5.0mL volumes Autoclave for 15 min at 15 psi pressure–121°C Cool to room temperature Aseptically add 0.88mL of ethanol solution to each tube containing 5.0mL of medium

Use: For the cultivation of Lactobacillus fructivorans.

Kupferberg Trichomonas Base

L-Cysteine·HCl·H2O 1.5g Agar 1.0g Maltose 1.0g Methylene Blue 3.0mg Bovine serum 50.0mL

pH 6.0 ± 0.2 at 25°C

Preparation of Medium: Add components, except bovine serum,

to distilled/deionized water and bring volume to 950.0mL Mix thor-oughly Gently heat and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C Aseptically add 50.0mL of bovine serum If desired, additional selectivity can be obtained by aseptically adding 250,000U of penicillin and 1.0g of streptomycin or 1.0g of chloramphenicol Mix thoroughly Pour into sterile Petri dishes or dis-tribute into sterile tubes

Use: For the cultivation of the Trichomonas species from clinical

specimens

Kupferberg Trichomonas Broth

Enzymatic digest of protein 20.0g

L-Cysteine·HCl·H2O 1.5g Agar 1.0g Maltose 1.0g Chloramphenicol 0.1g Methylene Blue 3.0mg Bovine serum 50.0mL

pH 6.0 ± 0.2 at 25°C

Preparation of Medium: Add components, except bovine serum,

to distilled/deionized water and bring volume to 950.0mL Mix thor-oughly Gently heat and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C Aseptically add bovine serum If desired, additional selectivity can be obtained by aseptically adding 250,000U penicillin and 1.0g streptomycin or 1.0g chloramphenicol Mix thoroughly Pour into sterile Petri dishes or distribute into sterile tubes

Use: For the cultivation of the Trichomonas species from clinical

speci-mens

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910 Kupferberg Trichomonas HiVeg Broth Base with Serum and Selective Supplement

Kupferberg Trichomonas HiVeg Broth Base

with Serum and Selective Supplement

(Trichomonas HiVeg Broth Base, Kupferberg)

Agar 1.0g

L-Cysteine·HCl 1.5g

Maltose 1.0g

Methylene Blue 3.0mg

Bovine serum 50.0mL

Selective supplement 10.0mL

pH 6.0 ± 0.2 at 25°C

Source: This medium, without bovine serum and selective

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

Selective Supplement Solution:

Penicillin 250,000U

Preparation of Selective Supplement Solution: Add penicilliln

to distilled/deionized water and bring volume to 10.0mL Mix

thor-oughly Filter sterilize

Preparation of Medium: Add components, except bovine serum

adn selective supplement, to distilled/deionized water and bring

vol-ume to 950.0mL Mix thoroughly Gently heat and bring to boiling

Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C

Aseptically add 50.0mL of bovine serum and 10.0mL selective

supple-ment

Use: For the cultivation of Trichomonas species from clinical specimens.

Agar 1.0g

L-Cysteine·HCl 1.5g

Maltose 1.0g

Methylene Blue 3.0mg

Bovine serum 50.0mL

Selective supplement 10.0mL

pH 6.0 ± 0.2 at 25°C

Source: This medium, without bovine serum and selective

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

Streptomycin 1.0g

Preparation of Selective Supplement Solution: Add

strepto-mycin to distilled/deionized water and bring volume to 10.0mL Mix

thoroughly Filter sterilize

Preparation of Medium: Add components, except bovine serum

and selective supplement, to distilled/deionized water and bring

vol-ume to 950.0mL Mix thoroughly Gently heat and bring to boiling

Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C

Aseptically add 50.0mL of bovine serum and 10.0mL selective

supple-ment

Use: For the cultivation of Trichomonas species from clinical

speci-mens

Plant hydrolysate 20.0g Agar 1.0g

L-Cysteine·HCl 1.5g Maltose 1.0g Methylene Blue 3.0mg Bovine serum 50.0mL Selective supplement 10.0mL

pH 6.0 ± 0.2 at 25°C

Source: This medium, without bovine serum and selective supple-ment, is available as a premixed powder from HiMedia

Chloramphenicol 1.0g

Preparation of Selective Supplement Solution: Add chloram-phenicol to distilled/deionized water and bring volume to 10.0mL Mix thoroughly Filter sterilize

Preparation of Medium: Add components, except bovine serum adn selective supplement, to distilled/deionized water and bring vol-ume to 950.0mL Mix thoroughly Gently heat and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C Aseptically add 50.0mL of bovine serum and 10.0mL selective supple-ment

Use: For the cultivation of Trichomonas species from clinical

speci-mens

Kushneria aurantia Medium

(DSMZ Medium 1195)

Composition per liter:

NaCl 78.0g MgSO4·7H2O 20.3g MgCl2·6H2O 13.0g Yeast extract 5.0g KCl 2.0g CaCl2·2H2O 0.33g NaBr 0.23g NaHCO3 0.06g

pH 7.3 ± 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 7.3 Distribute into tubes or flasks Gently heat while stirring and bring to boiling Mix thoroughly Autoclave for 15 min at 15 psi pressure– 121°C

Use: For the cultivation of Kushneria aurantia.

KVBA

KYE Agar

Agar 15.0g NaNO3 2.5g

KH2PO4 1.0g Yeast extract 1.0g MgSO4·7H2O 0.3g

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L Diphasic Blood Agar Medium 911

CaCl2·6H2O 0.15g

NaCl 0.1g

FeCl3 10.0mg

pH 10.0 ± 0.2 at 25°C

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

Use: For the cultivation of a variety of alkaliphilic bacteria

L Agar (Luria Agar)

Agar 15.0g

Yeast extract 5.0g

NaCl 0.5g

pH 7.0 ± 0.2 at 25°C

Glucose 10.0g

Preparation of Glucose Solution: Add glucose to

distilled/deion-ized water and bring volume to 100.0mL Mix thoroughly Filter

ster-ilize

Preparation of Medium: Add components, except glucose

solu-tion, to distilled/deionized water and bring volume to 980.0mL Mix

thoroughly Bring pH to 7.0 Gently heat and bring to boiling

Auto-clave for 15 min at 15 psi pressure–121°C Aseptically add 20.0mL of

sterile glucose solution Mix thoroughly Pour into sterile Petri dishes

or leave in tubes

Use: For the cultivation of Escherichia coli.

L Broth (Luria Broth)

NaCl 5.0g

Yeast extract 5.0g

Glucose 1.0g

pH 7.0 ± 0.2 at 25°C

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

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

L Broth DAP Thymidine

NaCl 5.0g

Yeast extract 5.0g

Diaminopimelic acid solution 10.0mL

Thymidine solution 10.0mL

pH 7.0 ± 0.2 at 25°C

Diaminopimelic Acid Solution:

Diaminopimelic acid 0.1g

Preparation of Diaminopimelic Acid Solution: Add diamin-opimelic acid to distilled/deionized water and bring volume to 10.0mL Mix thoroughly Filter sterilize

Thymidine Solution:

Thymidine 0.01g

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

Preparation of Medium: Add components, except diaminopimelic acid solution and thymidine solution, to distilled/deionized water and bring volume to 980.0mL Mix thoroughly Bring pH to 7.0 Autoclave for 15 min at 15 psi pressure–121°C Aseptically add 10.0mL of sterile diaminopimelic acid solution and 10.0mL of sterile thymidine solu-tion Mix thoroughly Aseptically distribute into sterile tubes or flasks

L Diphasic Blood Agar Medium (ATCC Medium 947)

Blood agar, diphasic base medium 700.0mL Rabbit blood, defibrinated 300.0mL Locke’s solution 150.0mL

pH 7.2–7.4 at 25°C

Blood Agar, Diphasic Base Medium:

Beef 25.0g Agar 10.0g Neopeptone 10.0g NaCl 2.5g

Preparation of Blood Agar, Diphasic Base Medium: Trim beef to remove fat Add 25.0g of lean beef to 250.0mL of distilled/de-ionized water Gently heat and bring to boiling Boil for 2–3 min Filter through Whatman #2 filter paper Add agar, neopeptone, and NaCl to filtrate Bring volume to 750.0mL with distilled/deionized water Mix thoroughly Adjust pH to 7.2–7.4 Gently heat and bring to boiling Au-toclave for 15 min at 15 psi pressure–121°C Cool to 50°–55°C

Locke’s Solution:

NaCl 8.0g Glucose 2.5g

KH2PO4 0.3g CaCl2 0.2g KCl 0.2g

Preparation of Locke's Solution: Add components to distilled/ deionized water and bring volume to 1.0L Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Cool to 50°–55°C

Preparation of Medium: Aseptically combine 700.0mL of sterile blood agar, diphasic base medium, with 300.0mL of sterile

defibrinat-ed rabbit blood warmdefibrinat-ed to 50°–55°C Mix thoroughly Aseptically dis-tribute 5.0mL volumes into 16 × 125mm screw-capped test tubes Allow to cool in a slanted position Overlay the agar in each tube with 1.0mL of sterile Locke’s solution

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912 L Diphasic Blood Agar Medium

Use: For the cultivation and maintenance of Trypanosoma species,

Leishmania donovani, Herpetomonas species, and Trypanosoma

neveulemairei.

L Diphasic Blood Agar Medium

(ATCC Medium 1011)

Blood agar, diphasic base medium 700.0mL

Rabbit blood, defibrinated 300.0mL

Locke’s solution 150.0mL

pH 7.2–7.4 at 25°C

Blood Agar, Diphasic Base Medium:

Beef 25.0g

Agar 10.0g

Neopeptone 10.0g

NaCl 2.5g

Preparation of Blood Agar, Diphasic Base Medium: Trim

beef to remove fat Add 25.0g of lean beef to 250.0mL of

distilled/de-ionized water Gently heat and bring to boiling Boil for 2–3 min Filter

through Whatman #2 filter paper Add agar, neopeptone, and NaCl to

filtrate Bring volume to 750.0mL with distilled/deionized water Mix

thoroughly Adjust pH to 7.2–7.4 Gently heat and bring to boiling

Au-toclave for 15 min at 15 psi pressure–121°C Cool to 50°–55°C

Locke’s Solution:

NaCl 8.0g

Glucose 2.5g

KH2PO4 0.3g

CaCl2 0.2g

KCl 0.2g

Preparation of Locke’s Solution: Add components to distilled/

deionized water and bring volume to 1.0L Mix thoroughly Autoclave

for 15 min at 15 psi pressure–121°C Cool to 50°–55°C

Preparation of Medium: Aseptically combine 700.0mL of sterile

blood agar, diphasic base medium, with 300.0mL of sterile

defibrinat-ed rabbit blood warmdefibrinat-ed to 50°–55°C Mix thoroughly Aseptically

dis-tribute 5.0mL volumes into 16 × 125mm screw-capped test tubes

Allow to cool in a slanted position Overlay the agar in each tube with

3.0mL of sterile Locke’s solution

Use: For the cultivation and maintenance of Trypanosoma species.

L and F Basal Salts, Modified with Heptadecane

NH4Cl 2.0g

Na2HPO4 0.21g

MgSO4·7H2O 0.2g

NaH2PO4 0.09g

KCl 0.04g

CaCl2 0.015g

FeSO4·7H2O 1.0mg

ZnSO4·7H2O 0.07mg

H3BO3 0.01mg

MnSO4·5H2O 0.01mg

MoO3 0.01mg

CuSO4·5H2O 5.0μg

Heptadecane 2.0mL

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components, except heptadecane, to distilled/deionized water and bring volume to 1.0L Mix thoroughly Gently heat and bring to boiling Distribute equally into four 250.0mL volumes Autoclave for 15 min at 15 psi pressure–121°C Cool to 60°C Filter sterilize heptadecane To one 250.0mL fraction of basal salts, aseptically add 0.5mL of sterile heptadecane Pour mixture into a sterile blender Homogenize slowly to mix heptadecane with basal salts and not to create excess bubbles Rapidly distribute medium to sterile screw-capped tubes Chill tubes quickly in an ice pack or in the refrig-erator Allow tubes to solidify in a slanted position

Use: For the cultivation and maintenance of Thermoleophilum album and Thermoleophilum minutum.

(ATCC Medium 167)

Na2HPO4 0.21g MgSO4·7H2O 0.2g NaH2PO4 0.09g KCl 0.04g CaCl2 0.015g FeSO4·7H2O 1.0mg Salts solution 1.0mL

Salts Solution:

ZnSO4·7H2O 7.0mg

H3BO3 1.0mg MnSO4·5H2O 1.0mg MoO3 1.0mg CuSO4·5H2O 0.5mg

Preparation of Salts Solution: Add components to distilled/deion-ized water and bring volume to 100.0mL Mix thoroughly

Use: For the cultivation and maintenance of Methylococcus

capsula-tus and Pseudomonas methanica.

L Medium (ATCC Medium 1154)

Pancreatic digest of casein 10.0g NaCl 5.0g Yeast extract 5.0g

pH 7.0 ± 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 7.0 Distribute into tubes or flasks Autoclave for 25 min at 15 psi pressure– 121°C

Use: For the cultivation and maintenance of Escherichia coli.

L Medium with Ampicillin

Pancreatic digest of casein 10.0g NaCl 5.0g

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L Medium with Methanol 913

Yeast extract 5.0g

Ampicillin solution 20.0mg

pH 7.0 ± 0.2 at 25°C

Ampicillin Solution:

Ampicillin 0.02g

Preparation of Ampicillin Solution: Add ampicillin to distilled/

deionized water and bring volume to 10.0mL Mix thoroughly Filter

sterilize

water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.0

Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–

121°C

L Medium with DAP and THY

(L Medium with Diaminopimelic Acid and Thymidine)

NaCl 5.0g

Yeast extract 5.0g

Diaminopimelic acid solution 10.0mL

Thymidine solution 10.0mL

pH 7.0 ± 0.2 at 25°C

D-Glucose 1.0g

Preparation of Glucose Solution: Add D-glucose to

distilled/de-ionized water and bring volume to 10.0mL Mix thoroughly Filter

ster-ilize

DL-Diaminopimelic acid 0.1g

Preparation of Diaminopimelic Acid Solution: Add

diamin-opimelic acid to distilled/deionized water and bring volume to 10.0mL

Mix thoroughly Filter sterilize

Thymidine 0.02g

Preparation of Thymidine Solution: Add thymidine to distilled/

deionized water and bring volume to 10.0mL Mix thoroughly Filter

sterilize

Preparation of Medium: Add components—except glucose

solu-tion, diaminopimelic acid solusolu-tion, and thymidine solution—to

dis-tilled/deionized water and bring volume to 970.0mL Mix thoroughly

Gently heat and bring to boiling Adjust pH to 7.0 Autoclave for 15

min at 15 psi pressure–121°C Cool to 45°–50°C Aseptically add

ster-ile glucose solution, diaminopimelic acid solution, and thymidine

so-lution Mix thoroughly Aseptically distribute into sterile tubes or

flasks

L Medium with DAP, THY, and AMP (L Medium with Diaminopimelic Acid, Thymidine, and Ampicillin)

Pancreatic digest of casein 10.0g NaCl 5.0g Yeast extract 5.0g Diaminopimelic acid solution 10.0mL Thymidine solution 10.0mL Ampicillin solution 10.0mL

pH 7.0 ± 0.2 at 25°C

DL-Diaminopimelic acid 0.1g

Preparation of Diaminopimelic Acid Solution: Add DL- di-aminopimelic acid to distilled/deionized water and bring volume to 10.0mL Mix thoroughly Filter sterilize

Thymidine 0.04g

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

Ampicillin Solution:

Ampicillin 0.02mg

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

Preparation of Medium: Add components—except ampicillin so-lution, diaminopimelic acid soso-lution, and thymidine solution—to dis-tilled/deionized water and bring volume to 970.0mL Mix thoroughly Gently heat and bring to boiling Adjust pH to 7.0 Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C Aseptically add ster-ile ampicillin solution, diaminopimelic acid solution, and thymidine solution Mix thoroughly Aseptically distribute into sterile tubes or flasks

L Medium with Diaminopimelic Acid and Thymidine

L Medium with Diaminopimelic Acid, Thymidine, and Ampicillin

L Medium with Methanol

Na2HPO4 0.21g MgSO4·7H2O 0.2g NaH2PO4 0.09g KCl 0.04g CaCl2 0.015g FeSO4·7H2O 1.0mg Methanol 20.0mL Salts solution 1.0mL

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914 L Medium for Salmonella

Salts Solution:

ZnSO4·7H2O 7.0mg

H3BO3 1.0mg

MnSO4·5H2O 1.0mg

MoO3 1.0mg

CuSO4·5H2O 0.5mg

Preparation of Salts Solution: Add components to

distilled/deion-ized water and bring volume to 100.0mL Mix thoroughly

Preparation of Medium: Add components, except methanol, to

distilled/deionized water and bring volume to 980.0mL Mix

thorough-ly Gently heat and bring to boiling Autoclave for 15 min at 15 psi

pressure–121°C Cool to 45°–50°C Filter sterilize methanol

Asepti-cally add 20.0mL of sterile methanol to cooled, sterile basal medium

Mix thoroughly Pour into sterile Petri dishes or distribute into sterile

tubes

Use: For the cultivation and maintenance of Methylobacillus

glyco-genes.

L Medium for Salmonella

NaCl 5.0g

Yeast extract 5.0g

Glucose 1.0g

pH 7.2 ± 0.2 at 25°C

wa-ter and bring volume to 1.0L Mix thoroughly Adjust pH to 7.2 Distribute

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

Use: For the cultivation and maintenance of Salmonella choleraesuis.

L Medium with Tetracycline

NaCl 5.0g

Yeast extract 5.0g

Tetracycline solution 10.0mL

pH 7.0 ± 0.2 at 25°C

Tetracycline Solution:

Tetracycline 0.02mg

Preparation of Tetracycline Solution: Add tetracycline to

dis-tilled/deionized water and bring volume to 10.0mL Mix thoroughly

Filter sterilize

Preparation of Medium: Add components, except tetracycline

so-lution, to distilled/deionized water and bring volume to 990.0mL Mix

thoroughly Gently heat and bring to boiling Adjust pH to 7.0

Auto-clave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C

Asepti-cally add sterile tetracycline solution Mix thoroughly AseptiAsepti-cally

distribute into sterile tubes or flasks

L15 Medium, Modified Leibovitz

NaCl 8.0g

DL-Threonine 0.6g

Sodium pyruvate 0.6g DL-Alanine 0.5g L-Arginine, free base 0.5g KCl 0.4g L-Asparagine·H2O 0.3g L-Histidine, free base 0.3g L-Glutamine 0.3g L-Isoleucine 0.3g L-Phenylalanine 0.3g L-Tyrosine 0.3g DL-Methionine 0.2g DL-Valine 0.2g Glycine 0.2g L-Serine 0.2g

Na2HPO4, anhydrous 0.2g CaCl2, anhydrous 0.1g L-Cysteine, free base 0.1g L-Leucine·HCl 0.1g MgCl2, anhydrous 0.094g D-Galactose 0.09g

KH2PO4 0.06g L-Tryptophan 0.02g Phenol Red 0.01g

i-Inositol 2.0mg

Choline chloride 1.0mg D-Calcium pantothenate 1.0mg Folic acid 1.0mg Nicotinamide 1.0mg Pyridoxine·HCl 1.0mg Thiamine monophosphate·2H2O 1.0mg Riboflavin-5-phosphate 0.1mg

pH 7.5 ± 0.2 at 25°C

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

at 5°C

Use: For the cultivation of oysters used for the growth of enterovi-ruses

L mono Confirmatory Agar Base

(Listeria monocytogenes Confirmatory Agar, Base)

Special peptone 30.0g Agar 12.0g LiCl 10.0g B.C indicator 8.6g Yeast extract 6.0g NaCl 5.0g α-Methyl-D-mannoside 3.0g

Na2HPO4, anhydrous 2.5g

Listeria mono enrichment supplement II 10.0mL Listeria mono selective supplement I 10.0mL Listeria mono selective supplement II 10.0mL

pH 7.2 ± 0.2 at 25°C

Source: This medium, without Listeria mono enrichment supplement

II, Listeria mono selective supplement I, and Listeria mono selective

supplement II, is available as a premixed powder from BioChemika

Caution: LiClis harmful Avoid bodily contact and inhalation of va-pors On contact with skin wash with plenty of water immediately

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