Handbook of Microbiological Media, Fourth Edition part 36 pot

Selective Supplement Solution: Composition per 10.0mL: Cefazolin...40.0mg Preparation of Selective Supplement Solution: Add cefazolin to distilled/deionized water and bring volume to 10.

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Charcoal Agar 345

Solution 1:

Composition per liter:

MgCl2·6H2O 40.0g

KCl 4.0g

CaCl2·2H2O 0.4g

pH 7.4 ± 0.2 at 25°C

Preparation of Solution 1: Add components to distilled/deionized

water and bring volume to 1.0L Mix thoroughly

Preparation of Medium: Add components to distilled/deionized

water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.4

Dis-tribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–

121°C

Use: For the cultivation of Haloarcula vallismortis.

Chaetomium Medium

Composition per liter:

Agar 15.0g

NaNO3 2.0g

K2HPO4 1.2g

MgSO4·7H2O 0.5g

KCl 0.5g

KH2PO4 0.14g

Yeast extract 0.02g

Fe2(SO4)3·H2O 0.01g

pH 7.2 ± 0.2 at 25°C

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 Allow

agar to cool Place a strip of sterile filter paper on top of agar and

inocu-late filter strip

Use: For the growth and maintenance of Chaetomium species.

Chalquist’s Antigen Medium, Modified

Soluble starch 0.5g

Pancreatic digest of casein 0.05g

L-Cysteine·HCl·H2O 0.01g

NAD (nicotinamide adeninedinucleotide) 0.01g

PPLO broth without Crystal Violet 90.0mL

Swine serum, inactivated 10.0mL

Phenol Red (1% solution) 0.25mL

pH 7.6 ± 0.2 at 25°C

PPLO Broth without Crystal Violet:

Composition per 500.0mL:

Beef heart, infusion from 11.52g

Peptone 2.32g

NaCl 1.15g

Source: PPLO broth without Crystal Violet is available as a premixed

powder from BD Diagnostic Systems

Preparation of PPLO Broth without Crystal Violet: Add

components to distilled/deionized water and bring volume to 500.0mL

Mix thoroughly

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 Mycoplasma synoviae.

Chapman Stone Agar Composition per liter:

(NH4)2SO4 75.0g NaCl 55.0g Gelatin 30.0g Agar 15.0g

D-Mannitol 10.0g Pancreatic digest of casein 10.0g

K2HPO4 5.0g Yeast extract 2.0g

pH 7.0 ± 0.2 at 25°C

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

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Autoclave for 10 min

at 15 psi pressure–121°C Pour into sterile Petri dishes while the

medi-um is still hot Add 25.0mL of medimedi-um per Petri dish

Use: For the isolation of staphylococci from a variety of specimens

Charcoal Agar Composition per liter:

Beef heart, solids from infusion 500.0g Agar 18.0g Peptone 10.0g Soluble starch 10.0g NaCl 5.0g Charcoal, activated, acid washed 4.0g Yeast extract 3.5g

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 Gently heat and bring

to boiling with frequent stirring Autoclave for 15 min at 15 psi pres-sure–121°C Cool to 45°–50°C Pour into sterile Petri dishes or leave

in tubes Shake flask while dispensing to keep charcoal in suspension Allow tubes to cool in a slanted position

Use: For the cultivation and maintenance of fastidious

microorgan-isms, especially Bordetella pertussis, for vaccine production.

Charcoal Agar Composition per liter:

Agar 12.0g Beef extract 10.0g Peptone 10.0g Starch 10.0g NaCl 5.0g Charcoal 4.0g Nicotinic acid 0.001g

pH 7.4 ± 0.2 at 25°C

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

to boiling with frequent stirring Autoclave for 15 min at 15 psi pres-sure–121°C Cool to 45°–50°C This medium may be enriched by the addition of blood Pour into sterile Petri dishes or distribute into tubes Shake flask while dispensing to keep charcoal in suspension

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346 Charcoal Agar Base, HiVeg with Blood and Selective Supplement

Use: For the cultivation and isolation of various bacteria; with the

addition of blood, for the cultivation of fastidious bacteria

Charcoal Agar Base, HiVeg

with Blood and Selective Supplement

Agar 18.0g

Plant infusion 12.0g

Plant peptone 10.0g

Starch 10.0g

NaCl 5.0g

Charcoal 4.0g

Yeast extract 3.5g

Sheep blood, defibrinated 50.0mL

Selective supplement solution 10.0mL

pH 7.3 ± 0.2 at 25°C

Source: This medium, without blood or selective supplement, is

available as a premixed powder from HiMedia

Selective Supplement Solution:

Cefazolin 40.0mg

Preparation of Selective Supplement Solution: Add cefazolin

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

thor-oughly Filter sterilize

Preparation of Medium: Add components, except blood and

se-lective supplement solution, to distilled/deionized water and bring

vol-ume to 940.0mL Mix thoroughly Gently heat and bring to boiling with

frequent stirring Autoclave for 15 min at 15 psi pressure–121°C Cool

to 45°–50°C Aseptically add 50.0 mL sterile sheep blood and 10.0mL

sterile selective supplement solution Mix thoroughly Pour into sterile

Petri plates or distribute into sterile tubes

addition of blood, for the cultivation of fastidious bacteria For the

cul-tivation of Bordetella pertussis for vaccine production and the

mainte-nance of stock cultures

Charcoal Agar with Horse Blood

Agar 12.0g

Beef extract 10.0g

Peptone 10.0g

Starch 10.0g

NaCl 5.0g

Charcoal, bacteriological 4.0g

Nicotinic acid 1.0mg

Horse blood, defibrinated 100.0mL

pH 7.4 ± 0.2 at 25°C

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

bring to boiling with frequent stirring Autoclave for 15 min at 15 psi

pressure–121°C Cool to 80°C Aseptically add 100.0mL of sterile,

defi-brinated horse blood Maintain at 80°C for 10 min to form chocolate

agar Pour into sterile Petri dishes or distribute into tubes Shake flask

while dispensing to keep charcoal in suspension

Use: For the cultivation and isolation of Haemophilus influenzae.

Charcoal Agar with Horse Blood and Cepahalexin Composition per liter:

Agar 12.0g Beef extract 10.0g Peptone 10.0g Starch 10.0g NaCl 5.0g Charcoal 4.0g Nicotinic acid 1.0mg Horse blood, defibrinated 100.0mL Cephalexin solution 10.0mL

pH 7.4 ± 0.2 at 25°C

Cephalexin Solution:

Cephalexin 0.04g

Preparation of Cephalexin Solution: Add cephalexin to dis-tilled/deionized water and bring volume to 10.0mL Mix thoroughly Filter sterilize

Preparation of Medium: Add components, except cephalexin so-lution and horse blood, to distilled/deionized water and bring volume

to 890.0L Mix thoroughly Gently heat and bring to boiling with fre-quent stirring Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C Aseptically add 100.0mL of sterile, defibrinated horse blood and 10.0mL of sterile cephalexin solution Pour into sterile Petri dishes or distribute into tubes Shake flask while dispensing to keep charcoal in suspension

Use: For the cultivation and isolation of Bordetella pertussis.

Charcoal Agar Slants

See: Diphasic Medium for Amoeba

Charcoal Blood Agar Base, HiVeg with Blood Penicillin

and Damidodiphenyl Hydrochloride Composition per liter:

Agar 12.0g Plant extract 10.0g Plant peptone 10.0g Starch 10.0g NaCl 5.0g Charcoal 4.0g Yeast extract 3.5g Sheep blood, defibrinated 50.0mL Penicillin solution 3.0mL Diamidodiphenylamine hydrochloride solution 3.0mL

pH 7.5 ± 0.2 at 25°C

Source: This medium, without blood or selective supplement, is available as a premixed powder from HiMedia

Penicllin Solution:

Penicillin 1000 units

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

Diamidodiphenylamine Hydrochloride Solution:

Diamido hydrochloride 0.01g Diphenylamine hydrochloride 0.01g

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CHCA Salts Medium 347

Preparation of Diamidodiphenylamine Hydrochloride

So-lution: Add components to distilled/deionized water and bring

vol-ume to 10.0mL Mix thoroughly Filter sterilize

Preparation of Medium: Add components, except blood,

penicil-lin solution, and diamidodiphenylamine hydrochloride solution, to

dis-tilled/deionized water and bring volume to 950.0mL Mix thoroughly

Gently heat and bring to boiling with frequent stirring Autoclave for

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

50.0 mL sterile sheep blood and 3.0 mL sterile penicillin solution and

3.0 mL sterile diamidodiphenylamine hydrochloride solution Mix

thoroughly Pour into sterile Petri plates or distribute into sterile tubes

addition of blood, for the cultivation of fastidious bacteria For the

cul-tivation of Bordetella pertussis for vaccine production and the

mainte-nance of stock cultures

Charcoal Blood Medium Composition per liter:

Beef heart, solids from infusion 500.0g

Agar 18.0g

Peptone 10.0g

Soluble starch 10.0g

NaCl 5.0g

Charcoal, activated, acid washed 4.0g

Yeast extract 3.5g

Horse or sheep blood, defibrinated 100.0mL

Cephalexin solution 10.0mL

pH 7.4 ± 0.2 at 25°C

Cephalexin Solution:

Cephalexin 0.04g

Preparation of Cephalexin Solution: Add cephalexin to

Filter sterilize

Preparation of Medium: Add components, except blood and

ce-phalexin solution, to distilled/deionized water and bring volume to

890.0mL 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 blood and cephalexin solution Mix thoroughly Pour into

sterile Petri dishes or distribute into sterile tubes

Use: For the cultivation of Haemophilus influenzae.

Charcoal HiVeg Agar Base

with Niacin, Blood, and Selective Supplement

Agar 12.0g

Plant extract 10.0g

Plant peptone No 2 10.0g

Starch 10.0g

NaCl 5.0g

Charcoal 4.0g

Niacin 1.0mg

Sheep blood, defibrinated 50.0mL

Selective supplement solution 10.0mL

pH 7.4 ± 0.2 at 25°C

Source: This medium, without blood or selective supplement, is

available as a premixed powder from HiMedia

Selective Supplement Solution:

Cefazolin 40.0mg

Preparation of Selective Supplement Solution: Add cefazolin

to distilled/deionized water and bring volume to 10.0mL Mix thor-oughly Filter sterilize

Preparation of Medium: Add components, except blood and se-lective supplement solution, to distilled/deionized water and bring vol-ume to 940.0mL Mix thoroughly Gently heat and bring to boiling with frequent stirring Autoclave for 15 min at 15 psi pressure–121°C Cool

to 45°–50°C Aseptically add 50.0 mL sterile sheep blood and 10.0mL sterile selective supplement solution Mix thoroughly Pour into sterile Petri plates or distribute into sterile tubes

Use: For the cultivation and isolation of Bordetella pertussis and Hae-mophilus influenzae For the cultivation of Bordetella pertussis for

vaccine production and the maintenance of stock cultures

Charcoal Yeast Extract Agar

Charcoal Yeast Extract Agar, Buffered

See: CYE Agar, Buffered Charcoal Yeast Extract Diphasic Blood Culture Medium

See: Legionella pneumophila Medium

Chase’s Medium SP Composition per liter:

Agar 10.0g Proteose peptone 10.0g

K2HPO4 2.0g (NH4)2SO4 1.0g Sucrose solution 100.0mL

pH 6.5 ± 0.2 at 25°C

Sucrose Solution:

Sucrose 10.0g

Preparation of Sucrose Solution: Add sucrose to distilled/deion-ized water and bring volume to 100.0mL Mix thoroughly Filter ster-ilize

Preparation of Medium: Add components, except sucrose solu-tion, to tap water and bring volume to 900.0mL Mix thoroughly Gently heat and bring to boiling Autoclave for 15 min at 15 psi pres-sure–121°C Cool to 45°–50°C Aseptically add 100.0mL of sterile sucrose solution Mix thoroughly Pour into sterile Petri dishes or dis-tribute into sterile tubes

Use: For the cultivation and maintenance of ATCC strain 13949

CHCA Salts Medium (Cyclohexane Carboxylic Acid Salts Medium) Composition per liter:

K2HPO4 3.5g

KH2PO4 1.5g Cyclohexane carboxylic acid 1.0g

NH4NO3 1.0g MgSO4·7H2O 0.5g FeSO4·7H2O 0.1g Yeast extract 0.1g

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348 Cheese Agar

CaCl2·2H2O 0.01g

Na2MoO2·2H2O 0.01g

ZnSO4·7H2O 0.01g

pH 7.0 ± 0.2 at 25°C

Gently heat and bring to boiling Distribute into tubes or flasks

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

or leave in tubes

Use: For the cultivation and maintenance of bacteria that can utilize

cyclohexane carboxylic acid as a carbon source For the cultivation and

maintenance of Arthrobacter globiformis

Cheese Agar Composition per liter:

Cheese, ripened 100.0g

NaCl 50.0g

Agar 15.0g

Peptone 10.0g

Potassium citrate 10.0g

Sodium oxalate 2.0g

pH 7.4 ± 0.2 at 25°C

Preparation of Medium: Add the cheese and potassium citrate to

distilled/deionized water and bring volume to 300.0mL Gently heat

and bring to 50°C to separate the fat Discard the fat In a separate flask,

add the remaining components to distilled/deionized water and bring

volume to 700.0mL Gently heat and bring to boiling Add the

300.0mL of aqueous suspension of cheese solids Adjust pH to 7.4

Au-toclave for 25 min at 15 psi pressure–121°C Pour into sterile Petri

dishes or distribute into sterile tubes

Use: For the isolation and cultivation of Brevibacterium linens.

Cherry Agar, CBS Formula

Pulp of sour stone cherries 200.0g

Agar 20.0g

pH 3.8 ± 0.2 at 25°C

Preparation of Medium: Add 1.0L of distilled/deionized water to

pulp of sour stone cherries Gently heat and bring to boiling Simmer

gently for 2 hr Strain through cloth Autoclave filtrate for 30 min at 6

psi pressure–110°C Add agar to distilled/deionized water and bring

volume to 800.0mL Gently heat and bring to boiling Autoclave for 15

min at 15 psi pressure–121°C To sterile agar solution, add 200.0mL of

sterile cherry filtrate Distribute into sterile tubes Autoclave for 5 min

at 15 psi pressure–121°C

Use: For the cultivation of various fungi

Cherry Decoction Agar Composition per liter:

Agar 20.0g

Sucrose 10.0g

Cherry extract 90.0mL

pH 3.8–4.6 at 25°C

Preparation of Medium: Add agar to distilled/deionized water and

bring volume to 500.0mL Gently heat and bring to boiling In a

sepa-rate flask, add sucrose and cherry extract to distilled/deionized water

and bring volume to 500.0mL Mix thoroughly Combine the two

solu-tions Adjust pH to 3.8–4.6 Distribute into tubes or flasks Sterilize by

tyndallization at 100°C for 30 min on 3 consecutive days Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation and maintenance of Acremonium sclerotigenum, Amylostereum laevigatum, Calcarisporium arbuscula, Helicoma mor-ganii, Helicoon richonis, Inonotus radiatus, Phellinus igniarius, Phoma leveillei, Taphrina californica, and Taphrina populina

CHI 1776 Medium Composition per liter:

Pancreatic digest of casein 25.0g Yeast extract 7.5g Glucose solution 25.0mL

Tris·HCl buffer, 1M, pH 7.5 20.0mL

Diaminopimelic acid solution 10.0mL Thymidine solution 10.0mL MgCl2 solution 5.0mL

Glucose Solution:

D-Glucose 20.0g

Preparation of Glucose Solution: Add glucose to distilled/deion-ized water and bring volume to 100.0mL Mix thoroughly Filter ster-ilize

Diaminopimelic Acid Solution:

meso-Diaminopimelic acid 1.0g

Preparation of Diaminopimelic Acid Solution: Add

meso-di-aminopimelic acid to distilled/deionized water and bring volume to 100.0mL Mix thoroughly Filter sterilize

Thymidine Solution:

Thymidine 0.4g

Preparation of Thymidine Solution: Add thymidine to distilled/ deionized water and bring volume to 100.0mL Mix thoroughly Filter sterilize

MgCl 2 Solution:

MgCl2 9.52g

Preparation of MgCl 2 Solution: Add MgCl2 to distilled/deion-ized water and bring volume to 100.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C

Preparation of Medium: Add components, except glucose solu-tion, diaminopimelic acid solusolu-tion, thymidine solusolu-tion, and MgCl2 so-lution, to distilled/deionized water and bring volume to 950.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C

Aseptical-ly add 25.0mL of sterile glucose solution, 10.0mL of sterile diamin-opimelic acid solution, 10.0mL of sterile thymidine solution, and 5.0mL of sterile MgCl2 solution, Mix thoroughly Distribute into sterile tubes or flasks

Use: For the cultivation of Escherichia coli.

Chicken Soup Broth Composition per 5.0L:

Chicken 2.5kg

Peppercorns 6

Cloves 3

Bay leaf 2

Celery, stalks including leaves 2

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Chitin Agar 349

Onion, large 1

Carrot 1

Dill, fresh 1/4 cup

NaCl 0.1g

Preparation of Medium: Add a nice, whole chicken to a large pot

Add enough tap water to cover the chicken by about 1 in Stud the

whole, peeled onion with the three cloves Add the onion and

remain-ing remain-ingredients to the pot Rapidly heat and brremain-ing to boilremain-ing Lower heat

to a simmer and cook for 1 to 1.5 hr Remove the chicken and

vegeta-bles from the broth Remove skin and bones from the chicken Cut up

the meat into 1-inch pieces Return the meat to the broth If desired,

slice the carrot and celery and return them to the broth

Use: For the growth and nutrition of microbiologists

China Blue Lactose Agar Composition per liter:

Agar 12.0g

Lactose 10.0g

Peptone 5.0g

NaCl 5.0g

Beef extract 3.0g

China Blue 0.375g

pH 7.0 ± 0.2 at 25°C

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

Unipath

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, differentiation, and enumeration of bacteria

from dairy products Lactose-fermenting bacteria appear as blue

colo-nies Nonlactose-fermenting bacteria appear as colorless colocolo-nies

China Blue Lactose Agar Composition per liter:

Agar 15.0g

Lactose 10.0g

Peptic digest of animal tissue 5.0g

NaCl 5.0g

Plant extract 3.0g

China Blue 0.3g

pH 7.0 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from

Hi-Media

Use: For the cultivation, differentiation, and enumeration of bacteria

from dairy products Lactose-fermenting bacteria appear as blue

colo-nies Nonlactose-fermenting bacteria appear as colorless colocolo-nies

China Blue Lactose HiVeg Agar

Agar 15.0g

Lactose 10.0g

Plant peptone 5.0g

NaCl 5.0g Plant extract 3.0g China Blue 0.3g

pH 7.0 ± 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 Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation, differentiation, and enumeration of bacteria from dairy products Lactose-fermenting bacteria appear as blue colo-nies Nonlactose-fermenting bacteria appear as colorless colocolo-nies

Chitin Agar Composition per liter:

Agar 15.0g Chitin, precipitated 3.0g (NH4)2SO4 2.0g

Na2HPO4 1.1g

KH2PO4 0.7g MgSO4·7H2O 0.2g FeSO4 1.0mg MnSO4 1.0mg

Chitin, Precipitated:

Composition per 2.5L:

Chitin 40.0g HCl, concentrated 400.0mL

Preparation of Chitin, Precipitated: Add chitin to 400.0mL of cold concentrated HCl Add this solution to 2.0L of distilled/deionized water at 5°C Filter the solution through Whatman #1 filter paper Dia-lyze the precipitated chitin against tap water for 12 hr Adjust the pH to 7.0 with KOH

Use: For the isolation and cultivation of Cytophaga species, Herpeto-siphon species, Saprospira species, and Flexithrix species.

Chitin Agar Composition per liter:

Agar 20.0g Chitin 4.0g

K2HPO4 0.7g MgSO4·7H2O 0.5g

KH2PO4 0.3g FeSO4·7H2O 0.01g MnCl2·4H2O 0.001g ZnSO4·7H2O 0.001g

pH 8.0 ± 0.2 at 25°C

Use: For the selective isolation and cultivation of streptomycetes

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350 Chlamydia Growth Medium

Chlamydia Growth Medium

Eagle minimum essential medium

with Earle salts, 10X 50.0mL

Fetal calf serum 50.0mL

L-Glutamine solution 5.0mL

pH 7.4 ± 0.2 at 25°C

Eagle Minimum Essential Medium with Earle Salts, 10X:

NaCl 6.8g

Glucose 1.0g

KCl 0.4g

CaCl2·2H2O 0.2g

MgCl2·6H2O 0.2g

NaH2PO4 0.15g

L-Arginine 0.1g

L-Lysine 0.06g

L-Isoleucine 0.05g

L-Leucine 0.05g

L-Threonine 0.05g

L-Valine 0.05g

L-Tyrosine 0.04g

L-Phenylalanine 0.03g

L-Histidine 0.03g

L-Cystine 0.02g

L-Methionine 0.02g

L-Tryptophan 0.01g

i-Inositol 2.0mg

Calcium pantothenate 1.0mg

Choline chloride 1.0mg

Folic acid 1.0mg

Nicotinamide 1.0mg

Pyridoxal 1.0mg

Thiamine·HCl 1.0mg

Riboflavin 0.1mg

Preparation of Eagle Minimum Essential Medium with

Earle Salts, 10X: Add components to distilled/deionized water and

bring volume to 1.0L Mix thoroughly Adjust pH to 7.4 with 7.5%

Na2CO3 solution Filter sterilize

Glutamine Solution:

Composition per 100.0mL:

L-Glutamine 2.92g

NaCl (0.85% solution) 100.0mL

Preparation of Glutamine Solution: Add the glutamine to the

0.85% NaCl solution Mix thoroughly Filter sterilize

Preparation of Medium: Aseptically combine 50.0mL of sterile

Ea-gle minimum essential medium with Earle salts, 10X, 50.0mL of fetal calf

serum, and 5.0mL of sterile glutamine solution Bring volume to 500.0mL

with sterile distilled/deionized water Mix thoroughly Aseptically

distrib-ute into sterile tubes or flasks

Use: For the cultivation of Chlamydia species.

Chlamydia Isolation Medium

Eagle minimum essential medium

with Earle salts, 10X 50.0mL

Fetal calf serum 50.0mL

Selective supplement 10.0mL

L-Glutamine solution 5.0mL

pH 7.4 ± 0.2 at 25°C

Eagle Minimum Essential Medium with Earle Salts, 10X: Composition per liter:

NaCl 6.8g Glucose 1.0g KCl 0.4g CaCl2·2H2O 0.2g MgCl2·6H2O 0.2g NaH2PO4 0.15g

L-Arginine 0.1g

L-Lysine 0.06g

L-Isoleucine 0.05g

L-Leucine 0.05g

L-Threonine 0.05g

L-Valine 0.05g

L-Tyrosine 0.04g

L-Phenylalanine 0.03g

L-Histidine 0.03g

L-Cystine 0.02g

L-Methionine 0.02g

L-Tryptophan 0.01g

i-Inositol 2.0mg

Calcium pantothenate 1.0mg Choline chloride 1.0mg Folic acid 1.0mg Nicotinamide 1.0mg Pyridoxal 1.0mg Thiamine·HCl 1.0mg Riboflavin 0.1mg

Preparation of Eagle Minimum Essential Medium with Earle Salts, 10X: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.4 with 7.5%

Na2CO3 solution Filter sterilize

Selective Supplement:

Glucose 0.594g Vancomycin 0.05g Gentamicin 0.01g Amphotericin B 2.0mg Cycloheximide 2.0mg

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

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

Glutamine Solution:

L-Glutamine 2.92g NaCl (0.85% solution) 100.0mL

Preparation of Glutamine Solution: Add the glutamine to the 0.85% NaCl solution Mix thoroughly Filter sterilize

Preparation of Medium: Aseptically combine 50.0mL of sterile Eagle minimum essential medium with Earle salts, 10X, 50.0mL of fe-tal calf serum, 10.0mL of selective supplement, and 5.0mL of sterile glutamine solution Bring volume to 500.0mL with sterile

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distilled/de-Chloramphenicol Ampicillin LB Medium 351

ionized water Mix thoroughly Aseptically distribute into sterile tubes

or flasks

Use: For the isolation and cultivation of Chlamydia species

Chlamydomonas Enriched Medium

Agar 15.0g

Sodium acetate, anhydrous 2.0g

Tryptose 2.0g

Yeast extract 2.0g

Use: For the cultivation of Chlamydomonas reinhardtii.

Chlamydomonas Mutant Agar

Agar 18.0g

K2HPO4 1.162g

Sodium acetate 1.0g

NaH2PO4 0.92g

Sodium citrate 0.5g

MgSO4·7H2O 0.3g

NH4NO3 0.3g

CaCl2 0.04g

FeCl3·6H2O 0.01g

Trace salts solution 10.0mL

pH 6.8 ± 0.2 at 25°C

Trace Salts Solution:

H3BO3 0.15g

LiSO4·4H2O 0.1g

MnSO4·4H2O 0.04g

CoCl2 0.02g

(NH4)6Mo7O24 0.015g

CuSO4·5H2O 0.004g

Preparation of Trace Salts Solution: Add components to

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

Gently heat and bring to boiling Distribute into tubes or flasks

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

or leave in tubes

Chlamydomonas Mutant Broth

K2HPO4 1.162g

Sodium acetate 1.0g

NaH2PO4 0.92g

Sodium citrate 0.5g

NH4NO3 0.3g

MgSO4·7H2O 0.3g

CaCl2 0.04g

FeCl3·6H2O 0.01g

Trace salts solution 10.0mL

pH 6.8 ± 0.2 at 25°C

Trace Salts Solution:

H3BO3 0.15g LiSO4·4H2O 0.1g MnSO4·4H2O 0.04g CoCl2 0.02g (NH4)6Mo7O24 0.015g CuSO4·5H2O 0.004g

Preparation of Trace Salts Solution: Add components to dis-tilled/deionized water and bring volume to 1.0L Mix thoroughly

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

Chlamydospore Agar Composition per liter:

Purified polysaccharide 20.0g Agar 15.0g

KH2PO4 1.0g (NH4)2SO4 1.0g Trypan Blue 0.1g Biotin 5.0μg

pH 5.1 ± 0.2 at 25°C

Use: For differentiating Candida albicans from other Candida species

on the basis of chlamydospore formation

Chloramphenicol Ampicillin LB Medium Composition per liter:

NaCl 10.0g Pancreatic digest of casein 10.0g Yeast extract 5.0g Ampicillin solution 10.0mL Chloramphenicol solution 10.0mL

pH 7.0 ± 0.2 at 25°C

Ampicillin Solution:

Ampicillin 40.0mg

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

Chloramphenicol Solution:

Chloramphenicol 5.0mg

Preparation of Chloramphenicol Solution: Add chlorampheni-col to distilled/deionized water and bring volume to 10.0mL Mix thor-oughly Filter sterilize

Preparation of Medium: Add components, except ampicillin solu-tion and chloramphenicol solusolu-tion, to distilled/deionized water and bring volume to 980.0mL Mix thoroughly Adjust pH to 7.0

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Auto-352 Chloramphenicol Erythromycin LB Medium

clave for 15 min at 15 psi pressure–121°C Aseptically add 10.0mL of

sterile ampicillin solution and 10.0mL of sterile chloramphenicol

solu-tion Mix thoroughly Aseptically distribute into sterile tubes or flasks

Use: For the cultivation and maintenance of Escherichia coli.

Chloramphenicol Erythromycin LB Medium

NaCl 10.0g

Yeast extract 5.0g

Erythromycin 10.0mL

Chloramphenicol 10.0mL

pH 7.0 ± 0.2 at 25°C

Erythromycin Solution:

Erythromycin 10mg

Preparation of Erythromycin Solution: Add erythromycin to

distilled/deionized water and bring volume to 10.0mL Mix

thorough-ly Filter sterilize

Preparation of Chloramphenicol Solution: Add

chlorampheni-col to distilled/deionized water and bring volume to 10.0mL Mix

Preparation of Medium: Add components, except erythromycin

so-lution and chloramphenicol soso-lution, to distilled/deionized water and

bring volume to 980.0mL Mix thoroughly Adjust pH to 7.0 Autoclave

for 15 min at 15 psi pressure–121°C Aseptically add 10.0mL of sterile

ampicillin solution and 10.0mL of sterile chloramphenicol solution Mix

thoroughly Aseptically distribute into sterile tubes or flasks

Chloramphenicol L Broth Medium No 1

NaCl 5.0g

Yeast extract 5.0g

Glucose 1.0g

Chloramphenicol solution 10.0mL

pH 7.0 ± 0.2 at 25°C

Preparation of Medium: Add components, except

chlorampheni-col solution, to distilled/deionized water and bring volume to 990.0mL

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

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

solu-tion and 10.0mL of sterile chloramphenicol solusolu-tion Mix thoroughly

Aseptically distribute into sterile tubes or flasks

Chloramphenicol L Broth Medium No 2 Composition per liter:

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

pH 7.0 ± 0.2 at 25°C

Preparation of Medium: Add components, except chlorampheni-col solution, to distilled/deionized water and bring volume to 990.0mL Mix thoroughly Adjust pH to 7.0 Autoclave for 15 min at 15 psi pres-sure–121°C Aseptically add 10.0mL of sterile chloramphenicol solu-tion and 10.0mL of sterile chloramphenicol solusolu-tion Mix thoroughly Aseptically distribute into sterile tubes or flasks

Chloramphenicol L Broth Medium No 3 Composition per liter:

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

pH 7.0 ± 0.2 at 25°C

Preparation of Medium: Add components, except chlorampheni-col solution, to distilled/deionized water and bring volume to 990.0mL Mix thoroughly Adjust pH to 7.0 Autoclave for 15 min at 15 psi pres-sure–121°C Aseptically add 10.0mL of sterile chloramphen icol solu-tion and 10.0mL of sterile chloramphenicol solusolu-tion Mix thoroughly Aseptically distribute into sterile tubes or flasks

Chloramphenicol LB Medium No.1 Composition per liter:

NaCl 10.0g Pancreatic digest of casein 10.0g Yeast extract 5.0g Chloramphenicol 50.0μg

Preparation of Medium: Adjust pH to 7.0 Autoclave at 15 psi pres-sure–121°C for 15 min For solid medium, add 15.0g of agar

Chloramphenicol LB Medium No 2 Composition per liter:

NaCl 10.0g Pancreatic digest of casein 10.0g

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Chlorella Broth without Glucose and Citrate 353

Yeast extract 5.0g

Chloramphenicol solution 10.0mL

pH 7.0 ± 0.2 at 25°C

Preparation of Medium: Add components, except

chlorampheni-col solution, to distilled/deionized water and bring volume to 990.0mL

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

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

10.0mL of sterile chloramphenicol solution Mix thoroughly

Asepti-cally distribute into sterile tubes or flasks

Chloramphenicol Yeast Glucose Agar

Glucose 20.0g

Agar 14.9g

Yeast extract 5.0g

Chloramphenicol 0.1g

pH 6.6 ± 0.2 at 25°C

Source: This medium is available from HiMedia

Use: For the selective enumeration of yeasts and molds in milk and

milk products It is also recommended by ISO Committee under the

specifications ISO 7954:1987

Chlorella Agar

Agar 17.0g

KNO3 2.5g

KH2PO4 2.45g

MgSO4·7H2O 2.4g

K2SO4 0.217g

FeSO4·7H2O 1.5mg

MnSO4·H2O 1.4mg

H3BO3 0.28mg

ZnSO4·7H2O 0.22mg

Na2MoO4·2H2O 0.05mg

CuSO4·5H2O 0.0078mg

Supplement solution 100.0mL

pH 4.5 ± 0.2 at 25°C

Supplement Solution:

Glucose 10.0g

Potassium citrate 32.0mg

Preparation of Supplement Solution: Add components to

Filter sterilize

Preparation of Medium: Add components, except supplement so-lution, to distilled/deionized water and bring volume to 900.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Cool to 50°C Aseptically add supplement solution Mix thoroughly Pour into Petri dishes or aseptically distribute into sterile tubes

Use: For the cultivation of Chlorella spp.

Chlorella Broth

KNO3 2.5g

KH2PO4 2.45g MgSO4·7H2O 2.4g

K2SO4 0.217g FeSO4·7H2O 1.5mg MnSO4·H2O 1.4mg

H3BO3 0.28mg ZnSO4·7H2O 0.22mg

Na2MoO4·2H2O 0.05mg CuSO4·5H2O 0.0078mg Supplement solution 100.0mL

pH 4.5 ± 0.2 at 25°C

Supplement Solution:

Glucose 10.0g Potassium citrate 32.0mg

Preparation of Supplement Solution: Add components to dis-tilled/deionized water and bring volume to 100.0mL Mix thoroughly Filter sterilize

Preparation of Medium: Add components, except supplement so-lution, to distilled/deionized water and bring volume to 900.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Cool to 50°C Aseptically add supplement solution Mix thoroughly Asepti-cally distribute into sterile tubes

Chlorella Broth without Glucose and Citrate

KNO3 2.5g

KH2PO4 2.45g MgSO4·7H2O 2.4g

K2SO4 0.217g FeSO4·7H2O 1.5mg MnSO4·H2O 1.4mg

H3BO3 0.28mg ZnSO4·7H2O 0.22mg

Na2MoO4·2H2O 0.05mg CuSO4·5H2O 0.0078mg Supplement solution 100.0mL

pH 4.5 ± 0.2 at 25°C

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

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354 Chlorinated Fatty Acid Medium

Chlorinated Fatty Acid Medium

(NH4)2SO4 0.5g

MgSO4·7H2O 0.1g

Ca(NO3)2 0.075g

2-Chloropropionate 0.54g

Phosphate buffer (20mM solution, pH 7.2) 1.0L

Trace elements solution 1.0mL

pH 7.2 ± 0.2 at 25°C

Trace Elements Solution:

FeSO4·7H2O 1.0g

MnSO4·H2O 1.0g

Co(NO3)2·6H2O 0.25g

CuCl2·2H2O 0.25g

Na2MoO4·2H2O 0.25g

ZnCl2 0.25g

H3BO3 0.1g

NH4VO3 0.1g

NiSO4·6H2O 0.1g

Preparation of Trace Elements Solution: Add components to

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

Preparation of Medium: Combine components Mix thoroughly

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

121°C

Use: For the cultivation of Alcaligenes species.

Chlorobiaceae Medium 1 Composition per 4990.0mL:

Solution 1 4.0L

O2-free water 860.0mL

NaHCO3 solution 100.0mL

Na2S·9H2O solution 20.0mL

Trace elements solution 5.0mL

Vitamin B12 solution 5.0mL

pH 6.8 ± 0.2 at 25°C

Solution 1:

Composition per 4.0L:

MgSO4·7H2O 2.5g

KCl 1.7g

KH2PO4 1.7g

NH4Cl 1.7g

CaCl2·2H2O 1.25g

Preparation of Solution 1: Add components to distilled/deionized

water and bring volume to 4.0L Mix thoroughly Autoclave for 45 min

at 15 psi pressure–121°C Cool to 25°C under 100% N2 Saturate with

CO2 by stirring under 100% CO2 for 30 min

O 2 -Free Water:

H2O 860.0mL

Preparation of O 2 -Free Water: Autoclave H2O for 15 min at 15

psi pressure–121°C Cool to 25°C under 100% N2

NaHCO 3 Solution:

NaHCO3 7.5g

Preparation of NaHCO 3 Solution: Add the NaHCO3 to distilled/ deionized water and bring volume to 100.0mL Mix thoroughly Gas with 100% CO2 for 20 min Filter sterilize with positive CO2 pressure

Na 2 S·9H 2 O Solution:

Na2S·9H2O 10.0g

Preparation of Na 2 S·9H 2 O Solution: Add Na2S·9H2O to dis-tilled/deionized water Mix thoroughly Gas with 100% N2 for 15 min

in a screw-capped bottle Tightly close cap Autoclave for 15 min at 15 psi pressure–121°C Cool to 25°C

Trace Elements Solution:

FeCl2·4H2O 1.5g CoCl2·6H2O 0.19g MnCl2·4H2O 0.1g ZnCl2 0.07g

H3BO3 0.06g NaMoO4·2H2O 0.04g CuCl2·2H2O 0.02g NiCl2·6H20 0.02g HCl (25% solution) 6.5mL

Preparation of Trace Elements 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 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

Preparation of Medium: To 4.0L of sterile, CO2-saturated solution

1, aseptically add the remaining components Mix thoroughly Adjust

pH to 6.8 Aseptically distribute into sterile 100.0mL bottles using pos-itive pressure of 95% N2 + 5% CO2 Completely fill bottles with

medi-um except for a pea-sized air bubble

Use: For the isolation and cultivation of members of the Chloro-biaceae

Chlorobiaceae Medium 2 Composition per 1051.0mL:

Solution 1 950.0mL

Na2S·9H2O solution 60.0mL NaHCO3 solution 40.0mL Vitamin B12 solution 1.0mL

pH 6.8 ± 0.2 at 25°C

Solution 1:

KH2PO4 1.0g

NH4Cl 0.5g MgSO4·7H2O 0.4g CaCl2·2H2O 0.05g Trace elements solution SL-8 1.0mL

Preparation of Solution 1: Add components to distilled/deionized water and bring volume to 950.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C

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