Handbook of Microbiological Media, Fourth Edition part 66 ppt

0.1g Ethanol, absolute...4.0mL Preparation of Cycloheximide Solution: Add components to distilled/deionized water and bring volume to 10.0mL.. 0.05g Preparation of Nalidixic Acid Solutio

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Enrichment Broth, pH 7.3 with Pyruvate 645

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 Prepare

freshly or boil and cool the medium just before use

Use: For the cultivation of both aerobic and anaerobic organisms For

the performance of sterility tests of turbid or viscous specimens

Enrichment Broth for Aeromonas hydrophila

Compositionper liter:

NaCl 5.0g

Maltose 3.5g

Yeast extract 3.0g

Bile salts No 3 1.0g

L-Cysteine·HCl·H2O 0.3g

Bromthymol Blue 0.03g

Novobiocin 5.0mg

pH 7.0 ± 0.2 at 25°C

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

Use: For the cultivation and enrichment of Aeromonas hydrophila.

Enrichment Broth, pH 7.3

Pancreatic digest of casein 17.0g

Yeast extract 6.0g

NaCl 5.0g

Papaic digest of soybean meal 3.0g

Glucose 2.5g

K2HPO4 2.5g

Nalidixic acid solution 8.0mL

Cycloheximide solution 5.1mL

Acriflavin·HCl solution 3.0mL

pH 7.3 ± 0.2 at 25°C

Cycloheximide Solution:

Compositionper 10.0mL:

Cycloheximide 0.1g

Ethanol, absolute 4.0mL

Preparation of Cycloheximide Solution: Add components to

distilled/deionized water and bring volume to 10.0mL Mix

thorough-ly Filter sterilize

Caution: Cycloheximide is toxic Avoid skin contact or aerosol

for-mation and inhalation

Nalidixic Acid Solution:

Nalidixic acid 0.05g

Preparation of Nalidixic Acid Solution: Add nalidixic acid to

Acriflavin·HCl Solution:

Acriflavin·HCl 0.05g

Preparation of Acriflavin·HCl Solution: Add acriflavin·HCl to

Preparation of Medium: Add components—except

cyclohexim-ide solution, nalidixic acid solution, and acriflavin·HCl solution—to

thorough-ly Gently heat and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C Aseptically add 5.1mL of sterile cycloheximide solution, 8.0mL of sterile nalidixic acid solution, and 3.0mL of sterile acriflavin·HCl solution Mix thoroughly Aseptically distribute into sterile tubes

Use: For the isolation, cultivation, and enrichment of a variety of microorganisms from nondairy foods

Enrichment Broth, pH 7.3 Compositionper liter:

Pancreatic digest of casein 17.0g Yeast extract 6.0g NaCl 5.0g Papaic digest of soybean meal 3.0g Glucose 2.5g

K2HPO4 2.5g Nalidixic acid solution 8.0mL Cycloheximide solution 5.1mL Acriflavin·HCl solution 2.0mL

pH 7.3 ± 0.2 at 25°C

Cycloheximide 0.1g Ethanol, absolute 4.0mL

Preparation of Cycloheximide Solution: Add components to distilled/deionized water and bring volume to 10.0mL Mix

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

Preparation of Nalidixic Acid Solution: Add nalidixic acid to distilled/deionized water and bring volume to 10.0mL Mix

Preparation of Acriflavin·HCl Solution: Add acriflavin·HCl to distilled/deionized water and bring volume to 10.0mL Mix

Preparation of Medium: Add components—except cyclohexim-ide solution, nalidixic acid solution, and acriflavin·HCl solution—to distilled/deionized water and bring volume to 984.9mL Mix

thorough-ly Gently heat and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C Aseptically add 5.1mL of cyclo-heximide solution, 8.0mL of nalidixic acid solution, and 2.0mL of acriflavin·HCl solution Mix thoroughly Aseptically distribute into sterile tubes

Use: For the isolation, cultivation, and enrichment of a variety of microorganisms from milk and dairy products

Enrichment Broth, pH 7.3 with Pyruvate Compositionper liter:

Pancreatic digest of casein 17.0g Yeast extract 6.0g

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646 Enrichment Broth, pH 7.3 with Pyruvate

NaCl 5.0g

Glucose 2.5g

K2HPO4 2.5g

Pyruvate solution 11.1mL

Nalidixic acid solution 8.0mL

Cycloheximide solution 5.1mL

Acriflavin·HCl solution 3.0mL

pH 7.3 ± 0.2 at 25°C

Pyruvate Solution:

Sodium pyruvate 2.0g

Preparation of Pyruvate Solution: Add sodium pyruvate to

dis-tilled/deionized water and bring volume to 20.0mL Mix thoroughly

Filter sterilize

Cycloheximide 0.1g

Ethanol, absolute 4.0mL

Preparation of Cycloheximide Solution: Add components to

Preparation of Nalidixic Acid Solution: Add nalidixic acid to

Preparation of Acriflavin·HCl Solution: Add acriflavin·HCl to

Preparation of Medium: Add components—except pyruvate

solu-tion, cycloheximide solusolu-tion, nalidixic acid solusolu-tion, and acriflavin·HCl

solution—to distilled/deionized water and bring volume to 972.8mL

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 11.1mL

of sterile pyruvate solution Mix thoroughly Inoculate medium and

in-cubate at 30°C for 6 hr Aseptically add 5.1mL of sterile cycloheximide

solution, 8.0mL of sterile nalidixic acid solution, and 3.0mL of sterile

acriflavin·HCl solution Mix thoroughly

Use: For the isolation, cultivation, and enrichment of Listeria species

from nondairy foods

Enrichment Broth, pH 7.3 with Pyruvate

NaCl 5.0g

Glucose 2.5g

K2HPO4 2.5g

Yeast extract 6.0g

Pyruvate solution 11.1mL Nalidixic acid solution 8.0mL Cycloheximide solution 5.1mL Acriflavin·HCl solution 2.0mL

pH 7.3 ± 0.2 at 25°C

Pyruvate Solution:

Sodium pyruvate 2.0g

Preparation of Pyruvate Solution: Add sodium pyruvate to dis-tilled/deionized water and bring volume to 20.0mL Mix thoroughly Filter sterilize

Cycloheximide 0.1g Ethanol, absolute 4.0mL

Preparation of Cycloheximide Solution: Add components to distilled/deionized water and bring volume to 10.0mL Mix

Preparation of Nalidixic Acid Solution: Add nalidixic acid to distilled/deionized water and bring volume to 10.0mL Mix

Preparation of Acriflavin·HCl Solution: Add acriflavin·HCl to distilled/deionized water and bring volume to 10.0mL Mix

Preparation of Medium: Add components—except pyruvate solu-tion, cycloheximide solusolu-tion, nalidixic acid solusolu-tion, and acriflavin·HCl solution—to distilled/deionized water and bring volume to 973.8mL 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 11.1mL

of sterile pyruvate solution Mix thoroughly Inoculate medium and in-cubate at 30°C for 6 hr Aseptically add 5.1mL of sterile cycloheximide solution, 8.0mL of sterile nalidixic acid solution, and 2.0mL of sterile acriflavin·HCl solution Mix thoroughly

Use: For the isolation, cultivation, and enrichment of Listeria species

from milk and dairy products

Entamoeba dispar Axenic

Culture Medium Compositionper liter:

YI-S solution 990.0mL Gastric mucin stock suspension 10.0mL

YI-S Solution:

YI broth 880.0mL Bovine serum, heat inactivated 100.0mL Vitamin mixture 18 20.0mL

Source: Vitamin mixture 18 is available from Biofluids, Rockville, MD

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Enteric Fermentation Base 647

YI Broth:

YI base stock 780.0mL

10× Glucose buffer stock 100.0mL

YI Base Stock:

Yeast extract 30.0g

L-Cysteine·HCl 1.0g

NaCl 1.0g

Ascorbic acid 0.2g

Ferric ammonium citrate 228.0mg

10× Glucose Buffer Stock:

Composition per 100.0.0mL:

Glucose 10.0g

K2HPO4 1.0g

KH2PO4 0.6g

Preparation of 10× Glucose Buffer Stock: Add components to

Preparation of YI Base Stock: Add components to 600.0mL of

distilled/deionized water Mix thoroughly Bring volume to 780.0mL

with distilled/deionized water Adjust pH to 6.8 with 1N NaOH

Dis-tribute in 78.0mL aliquots to 100.0mL screw-capped bottles

Auto-clave for 15 min at 15 psi pressure–121°C Cool to room temperature

Preparation of YI Broth: Aseptically add 10.0mL of 10× glucose

buffer stock to 78.0mL of cooled YI base stock Adjust osmolarity with

NaCl to 380.0milliosmols/kg

Preparation of YI-S Solution: Aseptically add 2.0mL of vitamin

mixture 18 and 10.0mL of heat-inactivated bovine serum to 88.0mL of

YI broth Distribute in 12.0mL aliquots to 16 x 125mm screw-capped

test tubes Store at 4°C in the dark with the caps screwed on tightly Use

within 96 hr

Gastric Mucin Stock Suspension:

Gastric mucin 1.0g

Preparation of Gastric Mucin Stock Suspension: Add gastric

mucin to distilled/deionized water and bring volume to 100.0mL Mix

thoroughly Autoclave for 15 min at 15 psi pressure–121°C

Preparation of Medium: Aseptically add 0.12mL of gastric mucin

stock suspension (thoroughly mix by swirling before use) to 12.0mL of

the complete YI-S solution in a 16 x 125mm screw-capped test tube

Store at 4°C in the dark with the caps screwed on tightly Use within 96

hrs

Use: For the cultivation of Entamoeba dispar

Entamoeba HiVeg Medium

Agar 11.0g

Plant infusion No 1 10.8g

Plant peptone No 3 5.5g

Sodium glycerophosphate 3.0g

NaCl 2.7g

pH 7.0 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from

Hi-Media

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 Allow the tubes to solidify in a slanted position Cover about half of the slant with fresh sterile horse seum saline mix-ture (1:6) and add a 5mm loopful of rice powder that has been sterilized

in an oven at 160°C Scorching should be prevented

Use: For the cultivation of Entamoeba histolytica.

Entamoeba Medium

(Endamoeba Medium) Compositionper liter:

Liver infusion 272.0g Rice powder 14.2g Agar 11.0g Proteose peptone 5.5g Sodium glycerophosphate 3.0g NaCl 2.7g Horse serum 50.0mL

pH 7.0 ± 0.2 at 25°C

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

Rice Powder:

Composition per 15.0g:

Rice powder 15.0g

Preparation of Rice Powder: Sterilize rice powder at 160°C for 60 min Do not overheat or rice powder will scorch

Preparation of Medium: Add components, except horse serum and rice powder, to distilled/deionized water and bring volume to 994.0mL Mix thoroughly Gently heat and bring to boiling Distribute into tubes in 7.0mL volumes Autoclave for 15 min at 15 psi pressure– 121°C Allow tubes to cool in a slanted position Aseptically add enough sterile horse serum to each tube to cover about half the slant Aseptically add 0.1g of sterile rice powder to each tube

Use: For the cultivation of Entamoeba histolytica.

Enteric Fermentation Base

(Fermentation Base for Campylobacter)

Peptic digest of animal tissue 10.0g NaCl 5.0g Beef extract 3.0g Carbohydrate solution 100.0mL Andrade’s indicator 10.0mL

pH 7.2 ± 0.1 at 25°C

Carbohydrate Solution:

Carbohydrate 10.0g

Preparation of Carbohydrate Solution: Add carbohydrate to distilled/deionized water and bring volume to 100.0mL Mix

thorough-ly Filter sterilize Glucose, lactose, mannitol, sucrose, adonitol, arabi-nose, cellobiose, dulcitol, glycerol, inositol, salicin, xylose, or other carbohydrates may be used For the preparation of expensive carbohy-drate solutions (adonitol, arabinose, cellobiose, dulcitol, glycerol, inos-itol, salicin, or xylose), 5.0g of carbohydrate per 100.0mL of distilled/ deionized water may be used

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648 Enterobacter Medium

Andrade’s Indicator:

NaOH (1N solution) 16.0mL

Acid Fuchsin 0.1g

Caution: Acid Fuchsin is a potential carcinogen and care must be

tak-en to avoid inhalation of the powdered dye and contact with the skin

Preparation of Andrade’s Indicator: Add components to

Preparation of Medium: Add components, except carbohydrate

solution, to distilled/deionized water and bring volume to 900.0mL

Mix thoroughly Gently heat and bring to boiling Distribute into tubes

that contain an inverted Durham tube in 9.0mL volumes Autoclave for

15 min at 15 psi pressure–121°C Cool to 25°C Aseptically add 1.0mL

of sterile carbohydrate solution per tube Mix thoroughly

Use: For the cultivation and differentiation of a variety of bacteria

based on their ability to ferment different carbohydrates Bacteria that

produce acid from carbohydrate fermentation turn the medium dark

pink to red Bacteria that produce gas have a bubble trapped in the

Dur-ham tube

Enterobacter Medium

Casein hydrolysate 2.0g

K2HPO4 1.4g

K2SO4 1.0g

Yeast extract 1.0g

KH2PO4 0.6g

MgSO4 0.5g

Glycerol 20.0mL

water and bring volume to 800.0mL Mix thoroughly Distribute into

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

Use: For the cultivation and maintenance of Enterobacter species and

Klebsiella pneumoniae.

Enterobacteria Enrichment Broth, Mossel

Pancreatic digest of gelatin 10.0g

Na2HPO4, anhydrous 6.4g

Glucose, anhydrous 4.5g

Bile salts (sodium desoxycholate, sodium lauryl sulphate,

sodium citrate) 3.055g

KH2PO4 2.0g

Brilliant Green 0.015g

pH 7.2 ± 0.2 at 25°C

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

Unipath

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

stirring and bring to boiling Maintain at 100°C for 30 min, typically

using flowing steam Do not autoclave Cool rapidly in cold running

water MixAseptically distribute into tubes or flasks

Use: For the enrichment of bile-tolerant Gram-negative bacteria in the

microbiological examination of pharmaceutical products

Enterobacteriaceae Enrichment Broth

See: EE Broth

Enterobacteriaceae Enrichment Broth, Mossel

See: EE Broth, Mossel Enterococci Broth, Chromocult (Chromocult Enterococci Broth) Compositionper liter:

Peptone 8.6g NaCl 6.4g Tween 80 2.2g NaN3 0.6g 5-Bromo-4-chloro-3-indolyl-β-D-glucopyranoside (X-GLU) 0.04

pH 7.5 ± 0.2 at 25°C

Source: This medium is available from Merck

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix well Distribute into tubes Auto-clave for 15 min at 15 psi pressure–121°C The prepared broth is clear and yellowish

Use: For the detection of enterococci in food and water The sodium azide present in this medium largely inhibits the growth of the accom-panying, and especially the Gram-negative microbial flora while spar-ing the enterococci The substrate X-GLU (5-bromo-4-chloro-3-indo-lyl-β-D-glucopyranoside) is cleaved, stimulated by selected peptones,

by the enzyme β-D-glucosidase which is characteristic for enterococci This results in an intensive blue-green color of the broth Azide, at the same time, prevents a false positive result by most other β-D -glucosi-dase positive bacteria Therefore, the color change of the broth largely confirms the presence of enterococci and group D streptococci

Enterococci Confirmatory Agar Compositionper liter:

Agar 15.0g Glucose 5.0g Pancreatic digest of casein 5.0g Yeast extract 5.0g NaN3 0.4g Methylene Blue 10.0mg Enterococci confirmatory broth variable

pH 8.0 ± 0.2 at 25°C

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

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

to boiling Distribute into tubes Autoclave for 15 min at 15 psi pres-sure–121°C Allow tubes to cool in a slanted position Add sufficient amount of Enterococci confirmatory broth (see below) to cover half the slant

Use: For the identification of enterococci from water by the confirma-tory test

Enterococci Confirmatory Broth Compositionper liter:

NaCl 65.0g Glucose 5.0g Pancreatic digest of casein 5.0g Yeast extract 5.0g NaN3 0.4g

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Enterococcus Confirmatory HiVeg Agar with Penicillin 649

Methylene Blue 10.0mg

Penicillin 650U

pH 8.0 ± 0.2 at 25°C

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

Di-agnostic Systems

Caution: Sodium azide is toxic Azides also react with metals and

disposal must be highly diluted

Preparation of Medium: Add components, except penicillin, to

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

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

pres-sure–121°C Cool to 25°C Aseptically add penicillin Mix thoroughly

Use: For the identification of enterococci from water by the

confirma-tory test

Enterococci Presumptive Broth

Glucose 5.0g

Yeast extract 5.0g

NaN3 0.4g

Bromthymol Blue 32.0mg

pH 8.4 ± 0.2 at 25°C

Di-agnostic Systems

Use: For the isolation and identification of enterococci from water by

the presumptive test Bacteria that produce acid and turn the medium

yellow and turbid after incubation at 45°C are presumptive

entero-cocci

Enterococcosel™ Agar Compositionper liter:

Agar 13.5g

Oxgall 10.0g

NaCl 5.0g

Yeast extract 5.0g

Peptic digest of animal tissue 3.0g

Esculin 1.0g

Sodium citrate 1.0g

Ferric ammonium citrate 0.5g

NaN3 0.25g

pH 7.1 ± 0.2 at 25°C

Di-agnostic Systems

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

stirring 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 rapid, selective isolation, cultivation, and enumeration of fecal group D streptococci (enterococci) For the cultivation of

staphy-lococci and Listeria monocytogenes.

Enterococcosel™ Broth Compositionper liter:

Pancreatic digest of casein 17.0g Oxgall 10.0g NaCl 5.0g Yeast extract 5.0g Peptic digest of animal tissue 3.0g Esculin 1.0g Sodium citrate 1.0g Ferric ammonium citrate 0.5g NaN3 0.25g

pH 7.1 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Gently heat while stirring until dissolved Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C

Use: For the cultivation and differentiation of group D streptococci (enterococci)

Enterococcus Agar

(m-Enterococcus Agar)

(Azide Agar) Compositionper liter:

Pancreatic digest of casein 15.0g Agar 10.0g Papaic digest of soybean meal 5.0g Yeast extract 5.0g

KH2PO4 4.0g Glucose 2.0g NaN3 0.4g Triphenyltetrazolium chloride 0.1g

pH 7.2 ± 0.2 at 25°C

to boiling Cool to 45°–50°C Do not autoclave Pour into sterile Petri dishes

Use: For the isolation, cultivation, and enumeration of entercocci in water, sewage, and feces by the membrane filter method For the direct plating of specimens for the detection and enumeration of fecal strep-tococci

Enterococcus Confirmatory HiVeg Agar

with Penicillin Compositionper liter:

Agar 15.0g Glucose 5.0g Plant hydrolysate 5.0g

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650 Enterococcus Confirmatory HiVeg Broth with Penicillin

Yeast extract 5.0g

NaN3 0.4g

Methylene Blue 0.01g

Penicillin solution 10.0mL

pH 8.0± 0.2 at 25°C

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

powder from HiMedia

Penicillin Solution:

Penicillin 650U

Preparation of Penicillin Solution: Add penicillin to

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

ster-ilize

Preparation of Medium: Add components, except penicillin

solu-tion, to distilled/deionized water and bring volume to 1.0L 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 10.0mL sterile

pen-icillin solution Mix thoroughly Pour into sterile Petri dishes or dispense

into sterile tubes

Use: For the isolation, cultivation, and enumeration of entercocci in

water, sewage, and feces by the membrane filter method For the direct

plating of specimens for the detection and enumeration of fecal

strep-tococci

Enterococcus Confirmatory HiVeg Broth

with Penicillin Compositionper liter:

NaCl 65.0g

Glucose 5.0g

Plant hydrolysate 5.0g

Yeast extract 5.0g

NaN3 0.4g

Methylene Blue 0.01g

Penicillin solution 10.0mL

pH 8.0 ± 0.2 at 25°C

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

powder from HiMedia

Penicillin 650U

Preparation of Penicillin Solution: Add penicillin to

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

ster-ilize

Preparation of Medium: Add components, except penicillin

solu-tion, to distilled/deionized water and bring volume to 1.0L 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 10.0mL sterile

pen-icillin solution Mix thoroughly

Use: For the cultivation of entercocci in water, sewage, and feces

Enterococcus faecium Medium

Sucrose 97.3g Brain heart, solids from infusion 37.0g Agar 13.3g NaCl 9.3g Yeast extract 5.0g MgSO4 0.25g

pH 7.4 ± 0.2 at 25°C

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 Enterococcus faecium.

Enterococcus Presumptive HiVeg Broth with Penicillin

Glucose 5.0g Plant hydrolysate 5.0g Yeast extract 5.0g NaN3 0.4g Bromthymol Blue 0.032g Penicillin solution 10.0mL

pH 8.4 ± 0.2 at 25°C

Source: This medium, without penicillin, is available as a premixed powder from HiMedia

Penicillin 650U

Preparation of Penicillin Solution: Add penicillin to distilled/de-ionized water and bring volume to 10.0mL Mix thoroughly Filter ster-ilize

Preparation of Medium: Add components, except penicillin solu-tion, to distilled/deionized water and bring volume to 1.0L 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 10.0mL sterile pen-icillin solution Mix thoroughly

Use: For the cultivation and presumptive identification of enterococci

Eosin Methylene Blue Agar

Eosin Methylene Blue Agar, Levine

See: Levine EMB Agar

Eosin Methylene Blue Agar, Modified

See: EMB Agar, Modified Epoxysuccinic Acid Medium

Erwinia amylovora Selective Medium

Agar 20.0g Mannitol 10.0g L-Asparagine 3.0g Sodium taurocholate 2.5g

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Erwinia tracheiphila Agar 651

K2HPO4 2.0g

Nicotinic acid 0.5g

MgSO4·7H2O 0.2g

Nitrilotriacetic acid 10.0mL

Actidione (cycloheximide) solution 10.0mL

Bromthymol Blue 9.0mL

Neutral Red 2.5mL

TlNO3 solution 1.75mL

Tergitol™ 7 0.1mL

pH 7.2–7.3 at 25°C

Cycloheximide 0.05g

Preparation of Cycloheximide Solution: Add cycloheximide to

TlNO 3 Solution:

TlNO3 0.1g

Preparation of TlNO 3 Solution: Add TlNO3 to

distilled/deion-ized water and bring volume to 10.0mL Mix thoroughly Filter

steril-ize

Preparation of Medium: Add components, except TlNO3 solution

and cycloheximide solution, to distilled/deionized water and bring

vol-ume to 988.25mL Mix thoroughly Adjust pH to 7.2–7.3 Gently heat

and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C

Cool to 45°–50°C Aseptically add 1.75mL of sterile TlNO3 solution

and 10.0mL of sterile cycloheximide solution Mix thoroughly Pour

into sterile Petri dishes or distribute into sterile tubes

Use: For the isolation and cultivation of Erwinia amylovora.

Erwinia Fermentation Medium

Lactose 45.0g

K2HPO4 3.6g

KH2PO4 1.8g

Yeast extract 1.8g

(NH4)2SO4 1.46g

MgSO4·7H2O 0.6g

CaCl2·2H2O 0.04g

FeSO4·7H2O 1.9mg

CoCl2 1.0mg

CuSO4·5H2O 1.0mg

MnSO4·H2O 1.0mg

Na2MoO4·2H2O 1.0mg

ZnSO4·7H2O 1.0mg

pH 7.0 ± 0.2 at 25°C

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

NaOH (approximately 0.18g) Distribute into tubes or flasks

Auto-clave for 15 min at 15 psi pressure–121°C

Use: For the cultivation of Rahnella aquatilis.

Erwinia Medium D3

Agar 15.0g

Arabinose 10.0g

Sucrose 10.0g LiCl 7.0g Casein hydrolysate 5.0g NaCl 5.0g MgSO4·7H2O 0.3g Acid Fuchsin 0.1g Bromthymol Blue 0.06g Sodium dodecyl sulfate 0.05g

pH 7.0 ± 0.2 at 25°C

Caution: Acid Fuchsin is a potential carcinogen and care must be

tak-en to avoid inhalation of the powdered dye and contact with the skin

to boiling Adjust pH to 8.2 Autoclave for 15 min at 15 psi pressure– 121°C Cool to 45°–50°C The pH after autoclaving should be 7.0 Pour into sterile Petri dishes or distribute into sterile tubes

Use: For the isolation and cultivation of Erwinia species

Erwinia Selective Medium

Agar 15.0g (NH4)2SO4 5.0g

K2HPO4 2.0g Eosin Y 0.4g Methylene Blue 0.065g Glycerol 10.0mL Antibiotic solution 10.0mL

Antibiotic Solution:

Cycloheximide 0.25g Novobiocin 0.04g Neomycin sulfate 0.04g

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

Preparation of Medium: Add components, except antibiotic solu-tion, to distilled/deionized water and bring volume to 990.0mL 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 sterile antibiotic solution Mix thoroughly Pour into sterile Petri dishes or distribute into sterile tubes

Use: For the selective isolation and cultivation of Erwinia species.

Erwinia tracheiphila Agar

Agar 15.0g Glucose 10.0g Peptone 10.0g Beef extract 5.0g Yeast extract 1.0g

pH 7.4 ± 0.2 at 25°C

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652 Erysipelothrix Medium

Use: For the cultivation and maintenance of Erwinia tracheiphila.

Erysipelothrix Medium

Na2HPO4·12H2O 18.0g

Glucose 6.0g

Peptone S 5.0g

Yeast extract 5.0g

L-Arginine·HCl 0.5g

Tween™ 80 0.5mL

pH 7.8–8.0 at 25°C

Use: For the cultivation of Erysipelothrix species.

Erythritol Agar Compositionper liter:

Agar 15.0g

Erythritol 2.0g

K2HPO4 1.15g

NH4NO3 1.0g

KH2PO4 0.625g

MgSO4·7H2O 0.02g

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 Klebsiella pneumoniae.

Erythritol Broth Compositionper liter:

Erythritol 2.0g

K2HPO4 1.15g

NH4NO3 1.0g

Yeast extract 1.0g

KH2PO4 0.625g

MgSO4·7H2O 0.02g

Use: For the cultivation of Klebsiella pneumoniae.

Erythrobacter longus Medium

Peptone 2.0g

Proteose peptone No 3 1.0g

Yeast extract 1.0g

Artificial seawater 700.0mL

Ferric citrate solution 2.0mL

pH 7.5 ± 0.2 at 25°C

Ferric Citrate Solution:

Ferric citrate 0.5g

Preparation of Ferric Citrate Solution: Add ferric citrate to

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

Use: For the cultivation of Erythrobacter longus.

Erythromicrobium roseococcus Medium

(DSMZ Medium 767) Composition per 1001.0mL:

Na-acetate 1.0g Yeast extract 1.0g Peptone 1.0g MgSO4·7H2O 0.5g

NH4Cl 0.3g

K2HPO4 0.3g KCl 0.3g CaCl2·2H2O 0.05g Trace elements solution SL-6 1.0mL Vitamin B12 solution 1.0mL

pH 7.5–7.8 at 25°C

Vitamin B 12 Solution:

Vitamin B12 2.0mg

Preparation of Vitamin B 12 Solution: Add vitamin B12 to dis-tilled/deionized water and bring volume to 100.0mL Mix thoroughly Filter sterilize

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

Preparation of Medium: Add components, except vitamin solution,

to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.5–7.8 Autoclave for 15 min at 15 psi pressure–121°C Cool to room temperature Aseptically add 10mL vitamin solution Mix thoroughly Aseptically distribute into sterile tubes or bottles

Use: For the cultivation of Erythrobacter litoralis, Erythromicrobium ramosum, and Roseococcus thiosulfatophilus.

Erythromycin L Broth Medium Compositionper liter:

Pancreatic digest of casein 10.0g NaCl 5.0g Yeast extract 5.0g Glucose 1.0g Erythromycin solution 10.0mL

pH 7.0 ± 0.2 at 25°C

Erythromycin Solution:

Erythromycin 10.0mg

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Escherichia Medium 653

Preparation of Erythromycin Solution: Add erythromycin to

Preparation of Medium: Add components, except erythromycin

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

pres-sure–121°C Aseptically add 10.0mL of sterile erythromycin solution

Mix thoroughly Aseptically distribute into sterile tubes or flasks

Use: For the cultivation of Escherichia coli.

Erythromycin LB Medium

NaCl 10.0g

Yeast extract 5.0g

Erythromycin solution 10.0mL

pH 7.0 ± 0.2 at 25°C

Erythromycin Solution:

Erythromycin 50.0mg

Preparation of Erythromycin Solution: Add erythromycin to

thor-oughly Filter sterilize

Preparation of Medium: Add components, except erythromycin

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

pres-sure–121°C Aseptically add 10.0mL of sterile erythromycin solution

Mix thoroughly Aseptically distribute into sterile tubes or flasks

ESA Medium (Epoxysuccinic Acid Medium)

Agar 20.0g

(NH4)2SO4 3.0g

KH2PO4 1.5g

Na2HPO4 1.5g

MgSO4·7H2O 0.5g

Yeast extract 0.5g

CaCl2·2H2O 0.01g

FeSO4·7H2O 0.01g

MnSO4·4H2O 0.001g

Epoxysuccinate solution 100.0mL

pH 7.0 ± 0.2 at 25°C

Epoxysuccinate Solution:

Sodium cis-epoxysuccinate 10.0g

Preparation of Epoxysuccinate Solution: Add sodium

cis-epox-ysuccinate to distilled/deionized water and bring volume to 100.0mL

Mix thoroughly Filter sterilize

Preparation of Medium: Add components, except epoxysuccinate

Mix thoroughly Gently heat and bring to boiling Autoclave for 15 min

at 15 psi pressure–121°C Cool to 50°–60°C Aseptically add sterile

epoxysuccinate solution Mix thoroughly Pour into sterile Petri dishes

or distribute into sterile tubes

Use: For the cultivation and maintenance of Nocardia tartaricans.

Escherichia coli Broth

See: EC Broth Escherichia Medium

(ATCC Medium 52) Compositionper liter:

Glucose 40.0g Agar 30.0g Peptone 10.0g

Use: For the cultivation and maintenance of Escherichia coli.

Escherichia Medium

(ATCC Medium 53) Composition per liter:

Agar 15.0g Casein hydrolysate 6.0g

K2HPO4 0.2g MgSO4·7H2O 0.2g Asparagine 0.15g FeSO4·7H2O 1.0μg Vitamin B12 solution 10.0mL Glycerol 2.0mL

pH 7.0 ± 0.2 at 25°C

Vitamin B 12 Solution:

Vitamin B12 0.04g

Preparation of Vitamin B 12 Solution: Add vitamin B12 to dis-tilled/deionized water and bring volume to 10.0mL Mix thoroughly

Preparation of Medium: Add components—except glycerol, agar, and vitamin B12 solution—to distilled/deionized water and bring vol-ume to 900.0mL Mix thoroughly Gently heat and bring to boiling Adjust pH to 7.0 Filter through Whatman #1 filter paper Add glycerol and agar Mix thoroughly Gently heat and bring to boiling Add vita-min B12 solution Distribute into tubes or flasks Autoclave for 15 min

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

Agar 15.0g

K2HPO4 7.0g Glycerol 5.0g

KH2PO4 3.0g (NH4)2SO4 1.5g L-Lysine 0.1g MgSO4 0.1g CaCl2 0.01g FeSO4·7H2O 0.5mg

pH 7.1 ± 0.2 at 25°C

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654 Escherichia Medium

Agar 15.0g

K2HPO4 7.0g

KH2PO4 3.0g

Sodium citrate, anhydrous 0.4g

MgSO4·7H2O 0.1g

(NH4)2SO4 0.1g

Glycerol 2.0mL

(ATCC Medium 62) Compositionper 700.0mL:

Agar 12.5g

Glycerol 2.0g

K2HPO4 0.2g

MgSO4·7H2O 0.2g

L-Asparagine 0.15g

FeSO4·7H2O 5.0mg

Vitamin B12 0.4mg

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components, except glycerol, agar,

and vitamin B12, to distilled/deionized water and bring volume to

500.0mL Adjust pH to 7.2 Mix thoroughly Gently heat and bring to

boiling Filter through Whatman #1 filter paper Cool to 25°C Add agar

and glycerol Bring volume to 1.0L with distilled/deionized water Mix

thoroughly Gently heat and bring to boiling Readjust pH to 7.2 Add

vi-tamin B12 Mix thoroughly Distribute into tubes in 10.0mL volumes

Autoclave for 15 min at 15 psi pressure–121°C Allow tubes to cool in a

slanted position

NaCl 8.0g

Yeast extract 1.0g

K2HPO4 11.67g Casamino acids 11.0g

KH2PO4 4.49g (NH4)2SO4 1.98g MgSO4·7H2O 0.25g FeSO4·7H2O 0.5mg Glycerol 20.25mL

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

Esculin Agar Compositionper liter:

Agar 15.0g Pancreatic digest of casein 13.0g NaCl 5.0g Yeast extract 5.0g Heart muscle, solids from infusion 2.0g Esculin 1.0g Ferric citrate 0.5g

pH 7.3 ± 0.2 at 25°C

to boiling Distribute into screw-capped tubes in 3.0mL volumes Au-toclave for 15 min at 15 psi pressure–121°C Allow tubes to cool in a slanted position

Use: For the cultivation and differentiation of bacteria based on their ability to hydrolyze esculin and produce H2S Bacteria that hydrolyze esculin appear as colonies surrounded by a reddish-brown to dark brown zone Bacteria that produce H2S appear as black colonies

Esculin Agar, Lombard-Dowell

See: Lombard-Dowell Esculin Agar

Esculin Agar, Modified CDC

(BAM M53) Composition per liter:

Heart muscle, infusion from 375.0g Agar 15.0g Thiotone 10.0g NaCl 5.0g Esculin 1.0g Ferric citrate 0.5g

pH 7.0 ± 0.2 at 25°C

to boiling Cool to 55°C Adjust pH to 7.0 Distribute into tubes or leave in flask Autoclave for 20 min at 15 psi pressure–121°C Pour into sterile Petri dishes or leave in tubes Allow tubes to cool in inclined position to produce slants

Use: For the differentiation of Enterobacter spp

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