Handbook of Microbiological Media, Fourth Edition part 138 pdf

3.0g Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L.. 10.0g Preparation of Medium: Add components to distilled/deionized water and bring volu

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Peptone Succinate Agar 1365

Peptone Recovery Broth

Composition per liter:

Meat extract 10.0g

Peptone 10.0g

NaCl 5.0g

pH 6.8 ± 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 6.8

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

121°C

Use: For the cultivation of Brevibacterium species.

Peptone-Reduced Agar

Agar 12.5g

Peptone 10.0g

Beef extract 5.0g

NaCl 3.0g

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 of Arthrobacter species.

Peptone Sodium Cholate

Meat extract 10.0g

Peptone 10.0g

NaCl 5.0g

Sodium cholate 5.0g

pH 6.8 ± 0.2 at 25°C

121°C

Use: For the cultivation and maintenance of Anthrobacter species.

Peptone Sorbitol Bile Broth

Sorbitol 10.0g

Na2HPO4 8.23g

NaCl 5.0g

Peptone 5.0g

Bile salts No 3 1.5g

NaH2PO4 1.2g

pH 7.6 ± 0.2 at 25°C

wa-ter and bring volume to 1.0L Mix thoroughly Distribute into bottles in

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

Use: For the enrichment and cultivation of Yersinia species.

Peptone Sorbitol HiVeg Broth

Sorbitol 10.0g

Na2HPO4 8.23g

Plant peptone 5.0g

NaCl 5.0g

Synthetic detergent No I 1.5g NaH2PO4 1.2g

pH 7.6 ± 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 wa-ter and bring volume to 1.0L Mix thoroughly Distribute into bottles in 100.0mL volumes Autoclave for 15 min at 15 psi pressure–121°C

Use: For the enrichment and cultivation of Yersinia species.

Peptone Starch Carbonate Medium

Composition per liter:

Agar 15.0g Soluble starch 10.0g Peptone 5.0g Yeast extract 5.0g

K2HPO4 1.0g MgSO4·7H2O 0.2g

Na2CO3 solution 100.0mL

Na 2 CO 3 Solution:

Composition per 100.0mL:

Na2CO3 10.0g

Preparation of Na 2 CO 3 Solution: Add Na2CO3 to distilled/de-ionized water and bring volume to 100.0mL Mix thoroughly Auto-clave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C

Preparation of Medium: Add components, except Na2CO3 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

Na2CO3 solution Mix thoroughly Pour into sterile Petri dishes or dis-tribute into sterile tubes

Use: For the cultivation and maintenance of Bacillus alcalophilus and other Bacillus species.

Peptone Starch Dextrose Agar

(PSD Agar) (Dunkelberg Agar)

Proteose peptone No 3 20.0g Agar 15.0g Soluble starch 10.0g Glucose 2.0g

Na2HPO4 1.0g NaH2PO4 1.0g

pH 6.8 ± 0.2 at 25°C

Preparation of Medium: Add starch to approximately 100.0mL of cold distilled/deionized water Mix thoroughly Add starch solution to 400.0mL of boiling distilled/deionized water Add remaining compo-nents Mix thoroughly Bring volume to 1.0L with distilled/deionized water Autoclave for 12 min at 8 psi pressure–112°C Pour into sterile Petri dishes or distribute into screw-capped tubes

Use: For the selective isolation and cultivation of Gardnerella

vagina-lis

Peptone Succinate Agar

Agar 15.0g Peptone 5.0g

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1366 Peptone Succinate Agar

Succinic acid 1.68g

MgSO4·7H2O 1.0g

(NH4)2SO4 1.0g

FeCl3·6H2O 2.0mg

MnSO4·H2O 2.0mg

pH 7.0 ± 0.2 at 25°C

Use: For the cultivation and maintenance of Aquaspirillum bengal,

Aquaspirillum dispar, and Spirillum volutans.

Peptone Succinate Agar

Peptone 5.0g

Succinic acid 1.68g

Agar 1.5g

MgSO4·7H2O 1.0g

NH4SO4 1.0g

FeCl3·6H2O 2.0mg

MnSO4·H2O 2.0mg

pH 7.0 ± 0.2 at 25°C

121°C

Use: For the cultivation of Aquaspirillum serpens.

Peptone Succinate Agar in Seawater

Peptone 5.0g

Succinic acid 1.68g

Agar 1.5g

MgSO4·7H2O 1.0g

(NH4)2SO4 1.0g

FeCl3·6H2O 2.0mg

MnSO4·H2O 2.0mg

Seawater 1.0L

pH 7.0 ± 0.2 at 25°C

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

Use: For the cultivation and maintenance of Oceanospirillum maris.

Peptone Succinate Salts Broth

(PSS Broth)

Peptone 1.0g

MgSO4·7H2O 0.1g

(NH4)2SO4 0.1g

Succinic acid 0.1g

FeCl3·6H2O 0.2mg

MnSO4·H2O 0.2mg

pH 6.8 ± 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 6.8 with KOH Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C

Use: For the cultivation of Spirillum species.

Peptone Succinate Salts Medium

(PSS Medium)

Peptone 10.0g MgSO4·7H2O 1.0g (NH4)2SO4 1.0g Succinic acid 1.0g FeCl3·6H2O 2.0mg MnSO4·H2O 2.0mg Synthetic seawater 1.0L

pH 6.8 ± 0.2 at 25°C

Synthetic Seawater:

NaCl 27.5g MgCl2 5.0g MgSO4 2.0g KCl 1.0g CaCl2 0.5g FeSO4 1.0μg

Preparation of Synthetic Seawater: Add components to dis-tilled/deionized water and bring volume to 1.0L Mix thoroughly

Preparation of Medium: Add solid components to 1.0L synthetic

seawater Mix thoroughly Adjust pH to 6.8 with 2N KOH Distribute

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

Use: For the cultivation and maintenance of Aquaspirillum anulus.

Peptone Succinate Salts in Seawater

Peptone 10.0g MgSO4·7H2O 1.0g (NH4)2SO4 1.0g Succinic acid 1.0g FeCl3·6H2O 2.0mg MnSO4·H2O 2.0mg Synthetic seawater 1.0L

pH 6.8 ± 0.2 at 25°C

Synthetic Seawater:

NaCl 27.5g MgCl2 5.0g MgSO4 2.0g KCl 1.0g CaCl2 0.5g FeSO4 1.0μg

Preparation of Synthetic Seawater: Add components to dis-tilled/deionized water and bring volume to 1.0L Mix thoroughly

Preparation of Medium: Add solid components to 1.0L synthetic

seawater Mix thoroughly Adjust pH to 6.8 with 2N KOH Distribute

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

Use: For the cultivation and maintenance of Oceanospirillum maris.

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Peptone Yeast Extract Agar 1367

Peptone Sucrose Broth

Sucrose 20.0g

Peptone 10.0g

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 maintenance of Xanthomonas campestris.

Peptone Water

Peptone 10.0g

NaCl 5.0g

pH 7.2 ± 0.2 at 25°C

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

Di-agnostic Systems and Oxoid Unipath

Use: For the cultivation of nonfastidious microorganisms, for

carbo-hydrate fermentation tests, and for performing the indole test

Peptone Water with Andrade’s Indicator

Peptone 10.0g

NaCl 5.0g

Andrade’s indicator 100.0mL

Carbohydrate solution 20.0mL

pH 7.4 ± 0.2 at 25°C

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

Unipath

Andrade’s Indicator:

NaOH (1N solution) 16.0mL

Acid Fuchsin 0.1 g

Preparation of Andrade’s Indicator: Add Acid Fuchsin to

NaOH solution and bring volume to 100.0mL with distilled/deionized

water

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

Carbohydrate Solution:

Carbohydrate 5.0–10.0g

Preparation of Carbohydrate Solution: Add carbohydrate to

distilled/deionized water and bring volume to 20.0mL Mix

thorough-ly Filter sterilize

Preparation of Medium: Add components, except carbohydrate

solution, to distilled/deionized water and bring volume to 980.0mL

Mix thoroughly Adjust pH to 7.4 if necessary Distribute into tubes

containing an inverted Durham tube Fill each tube with 9.8mL of

me-dium Autoclave for 15 min at 15 psi pressure–121°C Aseptically add

0.2mL of sterile carbohydrate solution to each tube

Use: For use in carbohydrate fermentation tests Fermentation is

deter-mined by the production of acid—broth turns pink—and formation of

gas—bubble trapped in Durham tube

Peptone Water, HiVeg

Plant peptone 10.0g NaCl 5.0g

pH 7.2 ± 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 Distribute into tubes

Use: For the cultivation of nonfastidious microorganisms, for carbohy-drate fermentation tests, and for performing the indole test Note: When sterile solutions are to be added after autoclaving, reduce the volume of water for reconstitution by an equal amount When used for carbohy-drate fermentation studies add inverted Durham tubes into the final con-tainers

Peptone Yeast Extract Agar (ATCC Medium 526)

Agar 15.0g Peptone 10.0g Yeast extract 3.0g

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 Bdellovibrio

bacteriovo-rus and Bdellovibrio stolpii.

Peptone Yeast Extract Agar (ATCC Medium 1093)

Use: For the cultivation and maintenance of Angiococcus disciformis.

Peptone Yeast Extract Agar

(ATCC 135)

Use: For the cultivation of Arthrobacter species.

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1368 Peptone Yeast Extract Agar

Peptone Yeast Extract Agar

(ATCC 1815)

Agar 20.0g

Glucose 5.0g

Peptone 5.0g

Yeast extract 3.0g

K2HPO4 0.2g

pH 7.0 ± 0.2 at 25°C

Preparation of Medium: Add components to tap water and bring

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

Ad-just pH to 7.0 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 Glycomyces tenuis.

Peptone Yeast Extract Carboxymethyl Cellulose Medium

Peptone Yeast Extract Glucose Agar

Agar 15.0g

Glucose 10.0g

Peptone 5.0g

Yeast extract 5.0g

pH 7.0 ± 0.2 at 25°C

Use: For the cultivation and maintenance of Alcaligenes latus,

Clavibacter iranicum, Clavibacter michiganense, Clavibacter rathayi,

Clavibacter tritici, Curtobacterium flaccumfaciens, Erwinia

amylo-vora, Erwinia mallotiamylo-vora, Erwinia nigrifluens, Erwinia quercina,

Erwinia rubrifaciens, Erwinia salicis, Gordona bronchialis, Gordona

terrae, Rhodococcus fascians, and Acinetobacter baumannii.

Peptone Yeast Extract Glucose Agar with Casein

Agar 15.0g

Glucose 10.0g

Peptone 5.0g

Yeast extract 5.0g

Casein hydrolysate 0.1g

pH 7.0 ± 0.2 at 25°C

Use: For the cultivation and maintenance of Clavibacter

michigan-ense.

Peptone Yeast Extract Glucose Broth

See: PYG Broth

Peptone Yeast Extract Glucose Maltose Medium

Peptone Yeast Extract Glucose Medium

Peptone Yeast Extract Glucose Medium for Spirillum

See: PYG Medium for Spirillum

Peptone Yeast Extract Glucose Medium, Modified

(MPYG Medium)

Peptone 10.0g Yeast extract 10.0g Glucose 5.0g

L-Cysteine·HCl·H2O 0.5g (NH4)2SO4 0.5g Salt solution 40.0mL Vitamin K-heme solution 10.0mL Resazurin (0.025% solution) 4.0mL Volatile fatty acid solution 3.1mL

pH 7.0 ± 0.2 at 25°C

Salt Solution:

NaHCO3 10.0g NaCl 2.0g

K2HPO4 1.0g

KH2PO4 1.0g CaCl2, anhydrous 0.2g MgSO4 0.2g

Preparation of Salt Solution: Add CaCl2 and MgSO4 to 300.0mL

of distilled/deionized water Mix thoroughly until dissolved Bring vol-ume to 800.0mL with distilled/deionized water Add remaining com-ponents while stirring Bring volume to 1.0L Mix thoroughly Store at 4°C

Vitamin K-Heme Solution:

Part A 100.0mL Part B 1.0mL

Preparation of Vitamin K-Heme Solution: Aseptically add 1.0mL of sterile part B to 100.0mL of cooled sterile part A Mix thor-oughly

Part A:

Hemin 50.0mg

Preparation of Part A: Add hemin to NaOH solution and bring volume to 100.0mL with distilled/deionized water Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C

Part B:

Menadione (vitamin K3) 100.0mg Ethanol (95% solution) 30.0mL

Preparation of Part B: Add menadione to ethanol Mix thoroughly Filter sterilize

Volatile Fatty Acid Solution:

Acetic acid 17.0mL Propionic acid 6.0mL

n-Butyric acid 4.0mL n-Valeric acid 1.0mL

Isovaleric acid 1.0mL

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Peptone Yeast Extract Medium 1369

Isobutyric acid 1.0mL

DL-α-Methyl butyric acid 1.0mL

Preparation of Volatile Fatty Acid Solution: Combine

compo-nents Mix thoroughly

Preparation of Medium: Add components—except vitamin

K-heme solution, L-cysteine·HCl·H2O, and volatile fatty acid solution—

to distilled/deionized water and bring volume to 936.9mL Gently heat

and bring to boiling under 97% N2 + 3% H2 Continue boiling until

re-sazurin turns colorless, indicating reduction Cool to 45°–50°C Add

vitamin K-heme solution, L-cysteine·HCl·H2O, and volatile fatty acid

solution Adjust pH to 7.0 Distribute into tubes under 97% N2 + 3%

H2 Cap with rubber stoppers Place tubes in a press Autoclave for 15

min at 15 psi pressure–121°C with fast exhaust

Use: For the cultivation and maintenance of Acetivibrio

ethanolgign-ens, Butyrivibrio fibrisolvethanolgign-ens, Lachnospira multipara, and

Succinivi-brio dextrinosolvens.

Peptone Yeast Extract Glucose Salt Medium

Peptone Yeast Extract Glucose Vitamin Marine Medium

See: PYGV Marine Medium

Peptone Yeast Extract Glucose Vitamin Medium

Peptone Yeast Extract Inositol Medium

See: PY Inositol Medium

Peptone Yeast Extract Iron Agar

See: ISP Medium 6

Peptone Yeast Extract Medium

(ATCC Medium 828)

Peptone 20.0g

Yeast extract 1.5g

pH 7.0 ± 0.2 at 25°C

psi pressure–121°C

Use: For the cultivation and maintenance of Acinetobacter lwoffii.

Peptone 10.0g

NaCl 5.0g

Yeast extract 5.0g

pH 7.2 ± 0.2 at 25°C

to boiling 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 Xenorhabdus

nematophi-lus

(PY Medium) (ATCC Medium 1524)

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

L-Cysteine·HCl·H2O 0.5g Salt solution 40.0mL Hemin solution 10.0mL Resazurin (0.025% solution) 4.0mL Vitamin K1 solution 0.2mL

pH 7.0 ± 0.2 at 25°C

Salt Solution:

NaHCO3 10.0g NaCl 2.0g

K2HPO4 1.0g

KH2PO4 1.0g CaCl2, anhydrous 0.2g MgSO4 0.2g

Preparation of Salt Solution: Add CaCl2 and MgSO4 to 300.0mL

of distilled/deionized water Mix thoroughly until dissolved Bring vol-ume to 800.0mL with distilled/deionized water Add remaining com-ponents while stirring Bring volume to 1.0L Mix thoroughly Store at 4°C

Hemin Solution:

Hemin 0.05g

Preparation of Hemin Solution: Add hemin to NaOH solution and bring volume to 100.0mL with distilled/deionized water Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C

Vitamin K1 Solution:

Vitamin K1 0.15g Ethanol (95% solution) 30.0mL

Preparation of Vitamin K1 Solution: Add vitamin K1 to ethanol Mix thoroughly Filter sterilize

Preparation of Medium: Add components—except vitamin K1 so-lution, hemin soso-lution, and L-cysteine·HCl·H2O, to distilled/deionized water and bring volume to 939.8mL Gently heat and bring to boiling under 80% N2 + 10% H2 + 10% CO2 Continue boiling until resazurin turns colorless, indicating reduction Cool to 45°–50°C Add vitamin

K1 solution, hemin solution, and L-cysteine·HCl·H2O Adjust pH to 7.0 Distribute into tubes under 80% N2 + 10% H2 + 10% CO2 Cap with rubber stoppers Place tubes in a press Autoclave for 15 min at 15 psi pressure–121°C with fast exhaust

Use: For the cultivation and maintenance of Megasphaera cerevisiae and Clostridium species.

Peptone Yeast Extract 1% Medium

Peptone Yeast Extract Medium with Fructose

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1370 Peptone Yeast Glutamate Medium

Peptone Yeast Extract Medium with Glucose

See: PY Medium with Glucose

Peptone Yeast Extract Salt Agar

See: PYS Agar

Peptone Yeast Extract Salt Medium

See: PY Salt Medium

Peptone Yeast Glutamate Medium

Peptone 20.0g

Yeast extract 10.0g

Monosodium glutamate 4.0g

Sodium thioglycolate 1.0g

pH 7.2 ± 0.2 at 25°C

Use: For the cultivation of Peptococcus aerogenes and a variety of

other bacteria

Peptone Yeast Medium with Magnesium Sulfate

(DSMZ Medium 790)

Peptone 10.0g

Yeast extract, dehydrated 1.0g

MgSO4·7H2O 2.0g

(NH4)2SO4 2.0g

pH 7.0 ± 0.2 at 25°C

Use: For the cultivation of Aquaspirillum psychrophilum and

Aqua-spirillum peregrinum subsp integrum.

Peptone Yeast Medium with MgSO4

Peptone 10.0g

MgSO4·7H2O 2.0g

(NH4)2SO4 2.0g

Yeast extract 1.0g

pH 7.0 ± 0.2 at 25°C

Use: For the cultivation and maintenance of Aquaspirillum itersonii,

Aquaspirillum peregrinum, and Aquaspirillum psychrophilum.

Peptone Yeast Trypticase™ Agar

Agar 15.0g

Peptone 6.0g

Trypticase™ (pancreatic digest of casein) 4.0g

Yeast extract 3.0g Beef extract 1.5g Glucose 1.0g

pH 7.0 ± 0.2 at 25°C

Use: For the cultivation of a variety of heterotrophic bacteria

Peptonized Milk Agar (PMA Medium)

Agar 15.0g Milk, peptonized 1.0g

Use: For the cultivation of freshwater Myxobacterium species.

Perfringens Agar, OPSP

See: Clostridium perfringens Agar, OPSP

Perfringens HiVeg Agar Base (O.P.S.P.)

with Antibiotics

Agar 15.0g Plant hydrolysate 15.0g Plant extract No 2 7.0g Papaic digest of soybean meal 5.0g Yeast extract 5.0g Tris buffer 1.5g Ferric ammonium citrate 1.0g

Na2S2O5 1.0g Antibiotic inhibitor 10.0mL

pH 7.3 ± 0.2 at 25°C

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

Antibiotic Inhibitor:

Sodium sulfadiazine 0.1g Oleandomycin phosphate 0.5mg Polymyxin B 10,000U

Preparation of Antibiotic Inhibitor: Add components to dis-tilled/deionized water and bring volume to 10.0mL Mix thoroughly Filter sterilize

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

Use: For the presumptive identification and enumeration of

Clostrid-ium perfringens in foods.

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Perkinsus Agar Medium 1371

Perfringens HiVeg Agar Base

with Egg Yolk and Antibiotics

(T.S.C./S.F.P HiVeg Agar Base)

Agar 15.0g

Plant hydrolysate No 1 15.0g

Papaic digest of soybean meal 5.0g

Plant extract 5.0g

Yeast extract 5.0g

Na2S2O5 1.0g

Ferric ammonium citrate 1.0g

Egg yolk emulsion 25.0mL

Perfringens SFP supplement 4.0mL

Perfringens TSC supplement 4.0mL

pH 7.6 ± 0.2 at 25°C

Source: This medium, without egg yolk emulsion, perfringens SFP

supplement, and perfringens TSC supplement, is available as a

pre-mixed powder from HiMedia

Egg Yolk Emulsion:

Composition per 100.0mL:

Chicken egg yolks 9

Whole chicken egg 1

NaCl (0.9% solution) 25.0mL

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 Beat to

form emulsion Measure 50.0mL of egg yolk emulsion and add to

50.0mL of 0.9% NaCl solution Mix thoroughly Filter sterilize Warm

to 45°–50°C

Perfringens SFP Supplement:

Kanamycin sulfate 30.0mg

Polymyxin B 75,000U

Preparation of Perfringens SFP Supplement: Add components

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

thor-oughly Filter sterilize

Perfringens TSC Supplement:

D-Cycloserine 1.0g

Preparation of Perfringens TSC Supplement: Add D

-cycloser-ine to distilled/deionized water and bring volume to 10.0mL Mix

thor-oughly Filter sterilize

Preparation of Medium: Add components, except perfringens SFP

supplement, egg yolk emulsion, and perfringens TSC supplement, to

distilled/deionized water and bring volume to 975mL Mix thoroughly

Gently heat and bring to boiling Distribute into tubes or flasks

Auto-clave for 15 min at 15 psi pressure–121°C Cool to 55°C Aseptically

add 25.0mL egg yolk emulsion, 4.0mL perfringens SFP supplement,

and 4.00mL perfringens TSC supplement Mix thoroughly Pour into

sterile Petri dishes or distribute into sterile tubes

Use: For the cultivation, enumeration, and presumptive identification

of Clostridium perfringens from foods.

Perkinsus Agar Medium (ATCC Medium 2289)

Modified Perkinsus Medium 485.0mL

Agar Medium 485.0mL

Fetal bovine serum, heat inactivated 20.0mL Lipid concentrate 10.0mL

Modified Perkinsus Medium Composition per liter:

HEPES 11.92g Nutrient mix F-12 Ham 10.8g Dulbecco’s modified Eagle’s medium 8.4g

L-Glutamine 0.29g NaHCO3 1.3g SASW 2X solution 860.0mL JPL carbohydrate solution 20.0mL Phenol Red (0.5%) solution 1.0mL

SASW 2X Solution:

Seawater, synthetic basal mixture 36.4g

Preparation of 2X SASW Solution: Add seawater synthetic basal mixture to distilled/deionized water and bring volume to 1.0L Mix thoroughly

JLP Carbohydrate Solution:

Glucose 5.0g Galactose 1.0g Trehalose 1.0g

Preparation of JLP Carbohydrate Solution: Add components

to distilled/deionized water and bring volume to 100.0mL Mix thor-oughly

Phenol Red Solution:

Phenol Red 0.05g

Preparation of Phenol Red Solution: Add Phenol Red to dis-tilled/deionized water and bring volume to 10.0mL

Preparation of Modified Perkinsus Medium: Add components

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

Agar Medium:

Agar 30.0g

Preparation of Agar Medium: Add agar to distilled/deionized wa-ter and bring volume to 1.0L Mix thoroughly Gently heat while stir-ring and bstir-ring to boiling Autoclave for 15 min at 15 psi pressure– 121°C Cool to 45°–50°C

Lipid Concentrate:

Pluronic™ F68 10.0g Tween™ 80 2.5g Cod liver oil 1.0g Cholesterol 0.45g

DL-α-Tocopherol acetate 0.2g

Preparation of Lipid Concentrate: Add components to distilled/ deionized water and bring volume to 1.0L Mix thoroughly Filter ster-ilize

Preparation of Medium: Warm Modified Perkinsus Medium to 50°C and combine with Agar Medium at 50°C Mix thoroughly and maintain at 50°C Aseptically add heat-inactivated fetal bovine serum and lipid concentrate Mix thoroughly Aliquot in 20mL amounts to Pe-tri dishes and allow to solidify

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1372 Perkinsus Medium

Use: For the cultivation of Perkinsus marinus, P andrewsi, P

chesa-peaki, and P atlanticus.

Perkinsus Medium

NaCl 9.0g

NaHCO3 2.1g

Glucose 1.1g

NaH2PO4·H2O 0.29g

KCl 0.38g

L-Arginine·HCl 0.21g

L-Glutamine 0.30g

MgSO4·7H2O 0.17g

Sodium pyruvate 0.11g

KH2PO4 0.08g

CaCl2·2H2O 0.09g

L-Cystine·2HCl 0.04g

L-Lysine·HCl 0.04g

L-Leucine 0.2g

L-Isoleucine 0.02g

L-Histidine·HCl·H2O 0.02g

L-Arginine 0.02g

L-Threonine 0.02g

L-Valine 0.02g

L-Tyrosine 0.02g

L-Methionine 0.02mg

L-Cystine 0.016g

L-Phenylalanine 0.014g

L-Serine 0.01g

L-Asparagine·H2O 0.01g

L-Aspartic Acid 0.01g

L-Glutamic acid 0.01g

L-Histidine 0.01g

L-Proline 0.01g

L-Glycine 0.01g

L-Alanine 8.9mg

D-Phenylalanine 5.0mg

L-Methionine 4.5mg

Hypoxanthine 4.1mg

L-Tryptophan 4.6mg

L-Threonine 3.6mg

L-Valine 3.5mg

L-Tyrosine 1.8mg

Vitamin B12 1.4mg

Folic acid 2.3mg

Phenol Red 1.2mg

Thiamine·HCl 2.0mg

FeSO4·7H2O 0.8mg

Choline chloride 1.7mg

Calcium DL-pantothenate 1.7mg

Thymidine 0.7mg

Niacinamide 1.6mg

Pyridoxal 1.0mg

Inositol 0.7mg

Riboflavin 0.5mg

Lipoic acid 0.2mg

Pyridoxine·HCl 0.2mg

ZnSO4·7H2O 0.03mg

FeNO3·7H2O 0.025mg

Biotin 0.02mg

CuSO4·5H2O 3.0μg

SASW solution 910.0mL

HEPES (N-[2-hydroxyethyl] piperazine-

N´-2-ethanesulfonic acid) buffer (1.0M solution) 25.0mL

Fetal bovine serum, heat inactivated 20.0mL JLP carbohydrate solution 10.0mL Lipid concentrate (100X) 10.0mL NaHCO3 solution 8.6mL

L-Glutamine solution 5.0mL Phenol Red solution 0.5mL

SASW Solution:

Seawater, synthetic basal mixture 18.2g

Preparation of SASW Solution: Add seawater synthetic basal mixture to distilled/deionized water and bring volume to 1.0L Mix thoroughly

JLP Carbohydrate Solution:

Glucose 5.0g Galactose 1.0g Trehalose 1.0g

Preparation of JLP Carbohydrate Solution: Add components

to distilled/deionized water and bring volume to 100.0mL Mix thor-oughly

Phenol Red Solution:

Phenol Red 0.05g

Preparation of Phenol Red Solution: Add Phenol Red to dis-tilled/deionized water and bring volume to 10.0mL

Glutamine Solution:

L-Glutamine 0.29g

Preparation of Glutamine Solution: Add L-glutamine to dis-tilled/deionized water and bring volume to 10.0mL

NaHCO 3 Solution:

NaHCO3 0.75g

Preparation of NaHCO 3 Solution: Add NaHCO3 to distilled/de-ionized water and bring volume to 10.0mL

Lipid Concentrate:

Pluronic™ F68 10.0g Tween™ 80 2.5g Cod liver oil 1.0g Cholesterol 0.45g

DL-α-Tocopherol acetate 0.2g

Preparation of Lipid Concentrate: Add components to distilled/ deionized water and bring volume to 1.0L Mix thoroughly Filter ster-ilize

Preparation of Medium: Add all components, except lipid concen-trate and fetal bovine serum, to distilled/deionized water and bring vol-ume to 1.0L Mix thoroughly Filter sterilize Aseptically add 20.0mL

of sterile fetal bovine serum and 10.0mL sterile lipid concentrate Mix thoroughly Aseptically distribute into sterile tubes or flasks Use im-mediately

Use: For the cultivation of Perkinsus marinus, P andrewsi, P

chesa-peaki, and P atlanticus.

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Petrotoga Medium 1373

Persephonella Medium

(DSMZ Medium 996)

NaCl 29.0g

MgSO4·7H2O 7.0g

NaOH 2.0g

Na2S2O3 2.0g

MgCl2·6H2O 1.36g

KCl 0.5g

CaCl2·2H2O 0.4g

K2HPO4 0.3g

NH4Cl 0.2g

Trace elements solution .10.0mL

pH 6.0 ± 0.2 at 25°C

Trace Elements Solution:

Na-EDTA·2H2O 0.5g

CoCl2·6H2O 0.15g

MnCl2·4H2O 0.1g

FeSO4·7H2O 0.1g

ZnCl2 0.1g

AlCl3·6H2O 40.0mg

Na2O4W·6H2O 30.0mg

CuCl 20.0mg

Ni2SO4·6H2O 20.0mg

Se-acide 10.0mg

H3BO3 10.0mg

Na2MoO4·2H2O 10.0mg

Preparation of Trace Elements Solution: Add components to

dis-tilled/deionized water and bring volume to 1.0L Adjust pH to 3.0 Mix

thoroughly

Preparation of Medium: Prepare anaerobic distilled/deionized

wa-ter by sparging with 100% CO2 Add components to distilled/deionized

anaerobic water and bring volume to 1.0L Adjust pH to 6.0 with

H2SO4 Autoclave for 15 min at 15 psi pressure–121°C Dispense

un-der a CO2 atmosphere into Bellco tubes (5mL medium per 27mL tube)

Stopper with butyl stoppers Cap and crimp closures Autoclave for 15

min at 15 psi pressure–121°C After autoclaving a white precipitate

might be present; this precipitate can be redissolved by shaking the

me-dium.It can take up to an hour before all precipitate is dissolved Add

3.8% O2 to each tube (1mL of O2 per 27mL tube) After inoculation

pressurize the tubes with H2 to 20psi (or 138kPa)

Use: For the cultivation of Persephonella spp.

Petragnani Medium

Skim milk 100.0g

Potato flour 36.4g

L-Asparagine 5.1g

Pancreatic digest of casein 5.1g

Malachite Green 1.2g

Whole egg 1277.0mL

Egg yolk 121.0mL

Glycerol 60.0mL

pH 7.0 ± 0.2 at 25°C

Source: This medium is available as a prepared medium from BD

Di-agnostic Systems

Preparation of Medium: Add components—except whole egg,

egg yolk, and glycerol—to distilled/deionized water and bring volume

to 940.0mL Mix thoroughly Add glycerol Gently heat while stirring and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C Scrub the eggshells with soap Let stand in a soap solution for 30 min Rinse in running water Soak eggs in 70% ethanol for 15 min Break the eggs into a sterile container Homogenize by shaking Filter through four layers of sterile cheesecloth into a sterile graduated cylinder Measure out 1277.0mL Add separated egg yolks

to another sterile container Measure out 121.0mL Aseptically add ho-mogenized whole egg and egg yolk to cooled sterile basal medium Mix thoroughly Aseptically distribute into sterile tubes Inspissate at 85°–90°C (moist heat) for 45 min

Use: For the isolation and cultivation of Mycobacterium species from clinical specimens For the cultivation and maintenance of

Mycobacte-rium smegmatis.

Petragnani Medium

Potato 500.0g Potato flour 36.0g Malachite Green 1.2g Whole egg 1200.0mL Whole milk 900.0mL Egg yolk 115.0mL Glycerol 70.0mL

pH 7.2 ± 0.2 at 25°C

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

Preparation of Medium: Peel and dice potato Add potato to 500.0mL of distilled/deionized water Gently heat and bring to boiling Continue boiling for 30 min Filter solids through two layers of cheese-cloth Combine potato solids with remaining components, except whole egg, egg yolk, and glycerol Mix thoroughly Add glycerol Gen-tly heat while stirring and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C Scrub the eggshells with soap Let stand in a soap solution for 30 min Rinse in running water Soak eggs in 70% ethanol for 15 min Break the eggs into a sterile container Homogenize by shaking Filter through four layers of sterile cheese-cloth into a sterile graduated cylinder Measure out 1200.0mL Add separated egg yolks to another sterile container Measure out 115.0mL Aseptically add homogenized whole egg and egg yolk to cooled sterile basal medium Mix thoroughly Aseptically distribute into sterile tubes Inspissate at 85°–90°C (moist heat) for 45 min

Use: For the isolation and cultivation of Mycobacterium species from clin-ical specimens For the cultivation and maintenance of Mycobacterium

smegmatis.

Petrotoga Medium

NaCl 18.0g MgSO4·7H2O 3.45g MgCl2·7H2O 2.75g NaHCO3 1.0g L-Cysteine·HCl·H2O 0.5g KCl 0.335g

NH4Cl 0.25g CaCl2·2H2O 0.14g

K2HPO4 0.14g Fe(NH4)2(SO4)2·7H2O 2.0mg Resazurin 1.0mg Glucose solution 50.0mL

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1374 Petrotoga Medium

Trace elements solution SL-6 10.0mL

Pancreatic digest of casein solution 10.0mL

Yeast extract solution 10.0mL

Na2S·9H2O solution 10.0mL

Wolfe’s vitamin solution 10.0mL

pH 6.5–6.7 at 25°C

Glucose Solution:

D-Glucose 5.0g

Preparation of Glucose Solution: Add glucose to

distilled/deion-ized water and bring volume to 50.0mL Mix thoroughly Sparge with

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

Trace Elements Solution SL-6:

H3BO3 0.3g

CoCl2·6H2O 0.2g

ZnSO4·7H2O 0.1g

MnCl2·4H2O 0.03g

Na2MoO4·H2O 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

Adjust pH to 3.4

Pancreatic Digest of Casein Solution:

Pancreatic digest of casein 1.0g

Preparation of Pancreatic Digest of Casein Solution: Add

pancreatic digest of casein to distilled/deionized water and bring

vol-ume to 10.0mL Mix thoroughly Sparge with 100% N2 Autoclave for

15 min at 15 psi pressure–121°C

Yeast Extract Solution:

Yeast extract 1.0g

Preparation of Yeast Extract Solution: Add yeast extract 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

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

Wolfe’s Vitamin Solution:

Pyridoxine·HCl 10.0mg

Thiamine·HCl 5.0mg

Riboflavin 5.0mg

Nicotinic acid 5.0mg

Calcium pantothenate 5.0mg

p-Aminobenzoic acid 5.0mg

Thioctic acid 5.0mg

Biotin 2.0mg

Folic acid 2.0mg

Cyanocobalamin 100.0μg

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

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2 Add components, except NaHCO3, glucose solution, pancreatic digest of casein solution, yeast extract solution, Na2S·9H2O solution, and Wolfe’s vitamin solution, to distilled/deionized water and bring volume to 910.0mL Mix thoroughly Adjust pH to 6.5–6.7 Gen-tly heat and bring to boiling Continue boiling for 3 min Cool to room temperature while sparging with 80% N2 + 20% CO2 Add NaHCO3 Adjust pH to 6.5–6.7 Anaerobically distribute 9.1mL volumes into an-aerobic tubes Autoclave for 15 min at 15 psi pressure–121°C Asepti-cally add 0.5mL of sterile glucose solution, 0.1mL of sterile pancreatic digest of casein solution, 0.1mL of sterile yeast extract solution, 0.1mL

of sterile Na2S·9H2O solution, and 0.1mL of sterile Wolfe’s vitamin so-lution to each tube Mix thoroughly

Use: For the cultivation of Petrotoga miotherma

Petrotoga Medium

(ATCC 1881)

NaCl 20.0g

Sodium PIPES

(piperazine-N,N´-bis[2-ethanesulfonic acid]) buffer 5.24g Pancreatic digest of casein 5.0g Yeast extract 2.0g Resazurin 1.0g Soluble starch 1.0g

L-Cysteine·HCl·H2O 0.5g

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components, except L -cyste-ine·HCl·H2O, to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.4 Gently heat and bring to boiling Continue boiling for 3 min Cool to room temperature while sparging with 100% N2 Add L-cysteine·HCl·H2O Mix thoroughly Anaerobi-cally distribute into tubes Autoclave for 15 min at 15 psi pressure– 121°C

Use: For the cultivation of Petrotoga miotherma

PFE Agar

See: Peptone Meat Extract Soil Extract Agar

Pfennig's Medium I, Modified for Marine Purple Sulfur Bacteria (DSMZ Medium 28)

Composition per 5.0L:

Solution A 4.0L Solution B 860.0mL Solution E 100.0mL Solution F 20.0mL Solution C 5.0mL Solution D 5.0mL

pH 7.3 at 25°C

Solution A:

Composition per 4.0L:

NaCl 100.0g MgSO4 15.0g

KH2PO4 1.7g

NH4Cl 1.7g

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