Handbook of Microbiological Media, Fourth Edition part 15 docx

Preparation of Medium: Add components—except Metals “44”, phosphate solution, and vitamin solution—to distilled/deionized water and bring volume to 1.0L.. Preparation of Medium: Add comp

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Aolpha Medium 135

Yeast extract 10.0g

Na2HPO4 1.0g

pH 6.0 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from

Hi-Media

Preparation of Medium: Add components to distilled/deionized

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

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

psi pressure–121°C Pour into sterile Petri dishes

Use: For for testing antimycotic sensitivity by the diffusion method

AO Agar

Composition per liter:

Agar 11.0g

Sodium acetate 0.5g

Pancreatic digest of casein 0.5g

Yeast extract 0.5g

Beef extract 0.2g

pH 7.2 ± 0.2 at 25°C

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

Use: For the isolation and cultivation of Cytophaga species,

Herpeto-siphon species, Saprospira species, and Flexithrix species.

AO Agar

Agar 4.0g

Sodium acetate 0.5g

Yeast extract 0.5g

Beef extract 0.2g

pH 7.2 ± 0.2 at 25°C

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

Use: For the maintenance of Cytophaga species, Herpetosiphon

spe-cies, Saprospira spespe-cies, and Flexithrix species.

AOAC Letheen Broth (Association of Official Analytical Chemists

Letheen Broth)

Peptic digest of animal tissue 10.0g

Polysorbate 80 5.0g

NaCl 5.0g

Beef extract 5.0g

Lecithin 0.7g

pH 7.0 ± 0.2 at 25°C

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

Di-agnostic Systems

to boiling Distribute into tubes in 10.0mL volumes Autoclave for 15

min at 15 psi pressure–121°C

Use: For the determination of phenol coefficients of disinfectant prod-ucts containing cationic surface-active materials Use according to

Official Methods of Analysis of the Association of Official Analytical Chemists (AOAC).

Aolpha Medium

Composition per 1041.0mL:

NaCl 100.0g Agar 15.0g MgSO4·7H2O 9.5g MgCl2·6H2O 5.0g KCl 5.0g Peptone 5.0g Yeast extract 1.0g CaCl2·2H2O 0.2g (NH4)2SO4 0.1g KNO3 0.1g Metals solution 20.0mL Phosphate solution 20.0mL Vitamin solution 1.0mL

pH 7.0 ± 0.2 at 25°C

Metals Solution:

Compositionper liter:

MgSO4·7H2O 29.7g Nitrilotriacetic acid 10.0g CaCl2·2H2O 3.3g FeSO4·7H2O 99.0mg

Na2MoO4·2H2O 12.7mg Metals “44” 50.0mL

Preparation of Metals Solution: Solubilize nitrilotriacetic acid with KOH Dissolve remaining ingredients Adjust pH to 7.2 with KOH or H2SO4 Autoclave for 15 min at 15 psi pressure–121°C Add aseptically to sterile basal medium

Metals “44”:

Compositionper 100.0mL:

ZnSO4·7H2O 1.1g FeSO4·7H2O 0.5g EDTA 0.25g MnSO4·7H2O 0.154g CuSO4·5H2O 0.04g Co(NO3)2·6H2O 0.025g

Na2B4O7·10H2O 0.018g

Preparation of Metals “44”: Add components to distilled/deion-ized water and bring volume to 100.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Add aseptically to sterile basal medium

Phosphate Solution:

K2HPO4 2.5g

KH2PO4 2.5g

Preparation of Phosphate Solution: Add components to dis-tilled/deionized water and bring volume to 1.0L Mix thoroughly Au-toclave for 15 min at 15 psi pressure–121°C Add aseptically to sterile basal medium

Vitamin Solution:

Pyridoxine·HCl 10.0mg Calcium pantothenate 5.0mg

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136 Aphanomyces Synthetic Medium

Nicotinamide 5.0mg

Riboflavin 5.0mg

Thiamine·HCl 5.0mg

Biotin 2.0mg

Folic acid 2.0mg

Cyanocobalamin 0.1mg

Preparation of Vitamin Solution: Add components to distilled/

deionized water and bring volume to 1.0L Mix thoroughly Filter

ster-ilize and add aseptically to sterile basal medium

Preparation of Medium: Add components—except Metals “44”,

phosphate solution, and vitamin solution—to distilled/deionized water

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

boiling Adjust pH of basal medium to 7.0 Autoclave for 15 min at 15

psi pressure–121°C Cool to 50°C and aseptically add the Metals “44”,

phosphate, and vitamin solutions

Use: For the cultivation and maintenance of Halomonas meridiana

and other Halomonas species.

Aphanomyces Synthetic Medium

D-Glucose 5.0g

KH2PO4 2.0g

L-Asparagine 0.75g

MgCl2 0.05g

FeCl3 5.0mg

MnCl2 5.0mg

ZnCl2 5.0mg

L-Methionine 0.02mg

pH 5.5 ± 0.2 at 25°C

water and bring volume to 1.0L Mix thoroughly Autoclave for 15 min

at 15 psi pressure–121°C Adjust pH to 5.5 Aseptically distribute into

sterile tubes or flasks

Use: For the cultivation of Aphanomyces species.

Aplanobacterium Medium

Agar 20.0g

Glucose 10.0g

Peptone 5.0g

Yeast extract 5.0g

pH 7.2 ± 0.2 at 25°C

to boiling Autoclave for 15 min at 15 psi pressure–121°C Pour into

sterile Petri dishes or leave in tubes

Use: For the cultivation and maintenance of Xanthomonas species.

Apple Juice Yeast Extract Medium

(AJYE Medium)

Agar 30.0g

Yeast extract 10.0g

Apple juice 1.0L

pH 4.8 ± 0.2 at 25°C

Preparation of Medium: Add yeast extract to 1.0L of apple juice

Mix thoroughly Adjust pH to 4.8 Autoclave for 10 min at 9 psi

pres-sure–114°C Cool to 45°–50°C In a separate flask, add agar to 200.0mL of distilled/deionized water and bring volume to 1.0L Mix thoroughly Gently heat and bring to boiling Autoclave for 15 min at

15 psi pressure–121°C Cool to 45°–50°C Aseptically combine the sterile apple juice solution with the sterile agar solution Mix

thorough-ly Pour into sterile Petri dishes

Use: For the cultivation of Zymomonas species.

APRY Agar

Fructose 30.0g NaCl 25.0g Glucose 20.0g Agar 15.0g Casein enzymatic hydrolysate 10.0g Yeast extract 2.5g Peptic digest of animal tissue 5.0g Acetic acid, glacial 5.0mL Selective supplement solution 5.0mL Potassium sorbate solution 1.0mL

pH 6.0 ± 0.2 at 25°C

Source: This medium is available from HiMedia

Potassium Sorbate Solution:

Potassium sorbate 1.0g

Preparation of Potassium Sorbate Solution: Add components

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

Selective Supplement Solution:

Chlortetracycline 50.0mg

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

Preparation of Medium: Add components, except acetic acid, se-lective supplement solution, and potassium sorbate solution, to dis-tilled/deionized water and bring volume to 990.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Cool to 50°C Asepti-cally add acetic acid, selective supplement solution, and potassium sor-bate solution Mix thoroughly Pour into Petri dishes or aseptically distribute into sterile tubes

Use: For the cultivation of acid resistant yeasts, including

Zygosac-charomyces bailii and ZygosacZygosac-charomyces rouxii in salads, sauces,

and dressings

APRY Agar Base with Acetic Acid and Sorbate

Fructose 30.0g NaCl 25.0g Glucose 20.0g Agar 15.0g Pancreatic digest of casein 10.0g Peptic digest of animal tissue 5.0g Yeast extract 2.5g Acetic acid (conc.) 5.0mL Potassium sorbate (10%) 1.0mL

pH 5.5 ± 0.2 at 25°C

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APT Agar 137

Source: This medium without acetic acid and potassium sorbate is

available as a premixed powder from HiMedia

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

50°C Aseptically add 5.0mL sterile acetic acid and 1.0mL potassium

sorbate solution Mix thoroughly Pour into sterile Petri dishes or

dis-tribute into sterile tubes

Use: For the detection and isolation of acid resistant yeasts,

Zygosac-charomyces bailii and ZygosacZygosac-charomyces rouxii, in salads, sauces,

and dressings

APRY Broth Base with Chloramphenicol

Glucose 30.0g

Fructose 20.0g

Polysorbate 80 10.0g

Yeast extract 2.5g

Choramphenicol solution 2.0ml

pH 6.5 ± 0.2 at 25°C

Source: This medium without chloramphenicol is available as a

pre-mixed powder from HiMedia Chloramphenicol supplement is

avail-able from HiMedia

Chloramphenicol Solution:

Chloramphenicol 50.0mg

Ethanol 2.0mL

Preparation of Chloramphenicol Solution: Add

chlorampheni-col to ethanol and bring volume to 2.0mL Mix thoroughly Filter

steril-ize

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

50°C Aseptically add 2.0mL chloramphenicol soultion Mix

thor-oughly Distribute into sterile tubes or flasks

Use: For the detection and isolation of acid resistant yeasts,

Zygosac-charomyces bailii and ZygosacZygosac-charomyces rouxii, in salads, sauces,

and dressings

APRY Broth

Glucose 30.0g

Fructose 20.0g

Casein enzymatic hydrolysate 15.0g

Yeast extract 2.5g

Polysorbate 80 10.0mL

Acetic acid, glacial 5.0mL

Selective supplement solution 5.0mL

Potassium sorbate solution 1.0mL

pH 6.0 ± 0.2 at 25°C

Potassium Sorbate Solution:

Potassium sorbate 1.0g

Preparation of Potassium Sorbate Solution: Add components

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

Chlortetracycline 50.0mg Chloramphenicol 50.0mg

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

Preparation of Medium: Add components, except acetic acid, se-lective supplement solution, and potassium sorbate solution, to dis-tilled/deionized water and bring volume to 990.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Cool to 50°C Asepti-cally add acetic acid, selective supplement solution, and potassium sor-bate solution Mix thoroughly Aseptically distribute into sterile tubes

or flasks

Use: For the cultivation of acid resistant yeasts, including

Zygosac-charomyces bailii and ZygosacZygosac-charomyces rouxii n salads, sauces, and

dressings

APT Agar

Agar 15.0g Pancreatic digest of casein 12.5g Glucose 10.0g Yeast extract 7.5g NaCl 5.0g

K2HPO4 5.0g Sodium citrate 5.0g

Na2CO3 1.25g MgSO4·7H2O 0.8g Polysorbate 80 0.2g MnCl2·4H2O 0.14g FeSO4·7H2O 0.04g Thiamine·HCl 1.0mg

pH 6.7 ± 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 Gently heat and bring

to boiling Distribute into tubes or flasks Autoclave for 15 min at 13 psi—118°–121°C Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation and enumeration of bacteria, especially heter-ofermentative lactobacilli, from meat and other foods For the cultiva-tion of streptococci

APT Agar

Agar 13.5g Pancreatic digest of casein 10.0g Glucose 10.0g Yeast extract 7.5g NaCl 5.0g

KH2PO4 5.0g Sodium citrate 5.0g

Na2CO3 1.25g MgSO4·7H2O 0.8g

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138 APT Broth

Polysorbate 80 0.2g

MnCl2·4H2O 0.14g

FeSO4·7H2O 0.04g

pH 6.7 ± 0.2 at 25°C

Di-agnostic Systems

psi—118°–121°C Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation and enumeration of bacteria, especially

heter-ofermentative lactobacilli, from meat and other foods For the

cultiva-tion of streptococci

APT Broth

Glucose 10.0g

Yeast extract 7.5g

NaCl 5.0g

K2HPO4 5.0g

Sodium citrate 5.0g

Na2CO3 1.25g

MgSO4·7H2O 0.8g

Polysorbate 80 0.2g

MnCl2·4H2O 0.14g

FeSO4·7H2O 0.04g

Thiamine·HCl 1.0mg

pH 7.7 ± 0.2 at 25°C

Di-agnostic Systems

psi—118°–121°C

Use: For the cultivation of lactic acid bacteria For the cultivation of

heterofermentative lactobacilli from meat and other foods The

Amer-ican Public Health Association recommends adding 100.0μg/L of

thi-amine to this medium

APT Broth

Glucose 10.0g

Yeast extract 7.5g

NaCl 5.0g

KH2PO4 5.0g

Sodium citrate 5.0g

Na2CO3 1.25g

MgSO4·7H2O 0.8g

Polysorbate 80 0.2g

MnCl2·4H2O 0.14g

FeSO4·7H2O 0.04g

pH 7.7 ± 0.2 at 25°C

Di-agnostic Systems

to boiling Distribute into tubes or flasks Autoclave for 15 min at 15 psi—118°–121°C

Use: For the cultivation of lactic acid bacteria For the cultivation of heterofermentative lactobacilli from meat and other foods The Amer-ican Public Health Association recommends adding 100.0μg/L of thi-amine to this medium

APT HiVeg Agar

Agar 15.0g Plant hydrolysate 12.5g Glucose 10.0g Yeast extract 7.5g NaCl 5.0g

KH2PO4 5.0g Sodium citrate 5.0g

Na2CO3 1.25g MgSO4·7H2O 0.8g Polysorbate 80 0.2g MnCl2·4H2O 0.14g FeSO4·7H2O 0.04g

pH 6.7 ± 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–121°C Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation and enumeration of bacteria, especially heter-ofermentative lactobacilli, from meat and other foods For the cultiva-tion of streptococci

APT HiVeg Broth

Plant hydrolysate 12.5g Glucose 10.0g Yeast extract 7.5g NaCl 5.0g

K2HPO4 5.0g Sodium citrate 5.0g

Na2CO3 1.25g MgSO4·7H2O 0.8g Polysorbate 80 0.2g MnCl2·4H2O 0.14g FeSO4·7H2O 0.04g Thiamine·HCl 1.0mg

pH 6.7 ± 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–121°C

Use: For the cultivation of lactic acid bacteria For the cultivation of heterofermentative lactobacilli from meat and other foods

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Aquaspirillum Autotrophic Broth 139

Aquabacter spiritensis Medium

(LMG Medium 225)

Sodium succinate 2.0g

Yeast extract 1.0g

KH2PO4 1.0g

Peptone 0.4g

NH4Cl 0.2g

NaCl 0.2g

MgSO4·7H2O 0.2g

CaCl2·2H2O 10.0mg

Ferric citrate 5.0mg

Vitamin solution 20.0mL

Trace elements solution SL-6 .1.0mL

pH 7.0 ± 0.2 at 25°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

compo-nents to distilled/deionized water and bring volume to 1.0L Mix

thor-oughly Adjust pH to 3.4

Vitamin Solution:

Calcium DL-pantothenate 5.0mg

Riboflavin 5.0mg

Thiamine·HCl 5.0mg

Biotin 2.0mg

Folic acid 2.0mg

Vitamin B12 0.1mg

Preparation of Vitamin Solution: Add components to distilled/

deionized water and bring volume to 1.0L Mix thoroughly Filter

ster-ilize

Preparation of Medium: Add components, except vitamin

solu-tion, to 980.0mL distilled/deionized water Mix thoroughly Autoclave

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

20.0mL sterile vitamin solution Mix thoroughly Aseptically distribute

to sterile tubes or flasks

Use: For the cultivation of Aquabacter spiritensis.

Aquaspirillum Autotrophic Agar

Noble agar 15.0g

Na2HPO4·12H2O 9.0g

KH2PO4 1.5g

NH4Cl 1.0g

MgSO4·7H2O 0.2g

CaCl2·2H2O 0.01g

Ferric ammonium citrate 5.0mg

NaHCO3 solution 10.0mL

Trace elements solution 3.0mL

pH 7.1 ± 0.2 at 25°C

NaHCO 3 Solution:

NaHCO3 0.5g

Preparation of NaHCO 3 Solution: Add the NaHCO3 to distilled/ deionized water and bring volume to 10.0mL Mix thoroughly Filter sterilize

Trace Elements Solution:

H3BO3 30.0mg CoCl2·6H2O 20.0mg ZnSO4·7H2O 10.0mg MnCl2·4H2O 3.0mg

Na2MoO4·2H2O 3.0mg NiCl2·6H2O 2.0mg CuCl2·2H2O 1.0mg

Preparation of Trace Elements Solution: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly

Preparation of Medium: Add components, except NaHCO3 solu-tion, to double-distilled 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 NaHCO3 so-lution Mix thoroughly Pour into sterile Petri dishes or distribute into sterile tubes For autotrophic growth, incubate under 85% H2 + 10% CO2 + 5% O2

Use: For the autotrophic cultivation and maintenance of

Aquaspiril-lum autotrophicum.

Aquaspirillum Autotrophic Broth

Na2HPO4·12H2O 9.0g

KH2PO4 1.5g

NH4Cl 1.0g MgSO4·7H2O 0.2g CaCl2·2H2O 0.01g Ferric ammonium citrate 5.0mg NaHCO3 solution 10.0mL Trace elements solution 3.0mL

pH 7.1 ± 0.2 at 25°C

NaHCO 3 Solution:

NaHCO3 0.5g

Preparation of NaHCO 3 Solution: Add the NaHCO3 to distilled/ deionized water and bring volume to 10.0mL Mix thoroughly Filter sterilize

H3BO3 30.0mg CoCl2·6H2O 20.0mg ZnSO4·7H2O 10.0mg MnCl2·4H2O 3.0mg

Na2MoO4·2H2O 3.0mg NiCl2·6H2O 2.0mg CuCl2·2H2O 1.0mg

Preparation of Trace Elements Solution: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly

Preparation of Medium: Add components, except NaHCO3 solu-tion, to double-distilled 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 NaHCO3 solu-tion Mix thoroughly Aseptically distribute into sterile tubes or flasks To grow autotrophically, incubate under 85% H2 + 10% CO2 + 5% O2

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140 Aquaspirillum Heterotrophic Agar

Use: For the autotrophic cultivation of Aquaspirillum autotrophicum.

Aquaspirillum Heterotrophic Agar

Noble agar 15.0g

Na2HPO4·12H2O 9.0g

KH2PO4 1.5g

NH4Cl 1.0g

MgSO4·7H2O 0.2g

CaCl2·2H2O 0.01g

pH 7.1 ± 0.2 at 25°C

H3BO3 30.0mg

CoCl2·6H2O 20.0mg

ZnSO4·7H2O 10.0mg

MnCl2·4H2O 3.0mg

Na2MoO4·2H2O 3.0mg

NiCl2·6H2O 2.0mg

CuCl2·2H2O 1.0mg

Preparation of Trace Elements Solution: Add components to

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

Preparation of Medium: Add components to double-distilled

wa-ter and bring volume to 1.0L Mix thoroughly Gently heat and bring to

boiling Autoclave for 15 min at 15 psi pressure–121°C Pour into

ster-ile Petri dishes or distribute into sterster-ile tubes

Use: For the heterotrophic cultivation and maintenance of

Aquaspiril-lum autotrophicum.

Aquaspirillum Heterotrophic Broth

Na2HPO4·12H2O 9.0g

KH2PO4 1.5g

NH4Cl 1.0g

MgSO4·7H2O 0.2g

CaCl2·2H2O 0.01g

pH 7.1 ± 0.2 at 25°C

H3BO3 30.0mg

CoCl2·6H2O 20.0mg

ZnSO4·7H2O 10.0mg

MnCl2·4H2O 3.0mg

Na2MoO4·2H2O 3.0mg

NiCl2·6H2O 2.0mg

CuCl2·2H2O 1.0mg

Preparation of Trace Elements Solution: Add components to

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

Preparation of Medium: Add components to double-distilled

wa-ter and bring volume to 1.0L Mix thoroughly Gently heat and bring to

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

distribute into sterile tubes or flasks

Use: For the heterotrophic cultivation of Aquaspirillum autotrophicum.

Aquaspirillum Medium

(DSMZ Medium 888)

Composition per 1026mL:

Casamino acids 1.5g (NH4)2SO4 1.0g MgSO4·7H2O 1.0g Agar 0.5g CaCl2·6H2O 30.0mg

Na2H2PO4 10.0mg Sodium succinate solution 10.0mL Thiosulfate solution 10.0mL Standard vitamin solution 5.0mL Trace elements solution SL-10 1.0mL

pH 7.5 ± 0.2 at 25°C

Standard Vitamin Solution:

Thiamine-HCl·2H2O 50.0mg Nicotinic acid 50.0mg Pyridoxine-HCl 50.0mg Ca-pantothenate 50.0mg Riboflavin 10.0mg Vitamin B12 1.0mg Folic acid 0.2mg Biotin 0.1mg

Preparation of Standard Vitamin Solution : Add components to

distilled/deionized water and bring volume to 100.0mL Mix

thorough-ly Filter sterilize

Trace Elements Solution SL-10:

FeCl2·4H2O 1.5g

H3BO3 300.0mg CoCl2·6H2O 190.0mg MnCl2·4H2O 100.0mg ZnCl2 70.0mg

Na2MoO4·2H2O 36.0mg NiCl2·6H2O 24.0mg CuCl2·2H2O 2.0mg HCl (25% solution) 7.7mL

Preparation of Trace Elements Solution SL-10: Add FeCl2·4H2O to 10.0mL of HCl solution Mix thoroughly Add distilled/ deionized water and bring volume to 1.0L Add remaining compo-nents Mix thoroughly Sparge with 100% N2 Autoclave for 15 min at

15 psi pressure–121°C Cool to room temperature

Thiosulfate Solution:

Na2S2O3·5H2O 1.0g

Preparation of Thiosulfate Solution: Add Na2S2O3·5H2O to dis-tilled/deionized water and bring volume to 10.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Cool to room temper-ature

Sodium Succinate Solution:

Preparation of Sodium Succinate Solution: Add sodium succi-nate to distilled/deionized water and bring volume to 10.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Cool to room temperature

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ARC51 Medium 141

Preparation of Medium: Add components, except sodium

succi-nate solution, thiosulfate solution, standard vitamin solution, and trace

elements solution SL-10, to distilled/deionized water and bring volume

to 1.0L Mix thoroughly Autoclave for 15 min at 15 psi pressure–

121°C Cool to room temperature Aseptically add 10.0mL sterile

so-dium succinate solution, 10.0mL sterile thiosulfate solution, 5.0mL

sterile standard vitamin solution, and 1.0mL sterile trace elements

so-lution SL-10 Mix thoroughly Adjust pH to 7.5 Aseptically distribute

into sterile tubes or flasks

Use: For the cultivation of Aquaspirillum spp.

Aquincola Medium

Yeast extract 1.0g

Peptone 1.0g

Fructose 0.5g

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

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

Use: For the cultivation and maintenance of Aquincola spp.

Aquisalimonas Agar

Solution A 500.0mL

Solution B 500.0mL

pH 9.5 ± 0.2 at 25°C

Solution A:

Agar 20.0g

Glucose 10.0g

Peptone 5.0g

Yeast extract 5.0g

K2HPO4 1.0g

MgCl2·6H2O 0.2g

Preparation of Solution A: Add components to distilled/deionized

water and bring volume to 500.0mL Mix thoroughly Autoclave for 15

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

Solution B:

NaCl 80.0g

Na2CO3 20.0g

Preparation of Solution B: Add components to distilled/deionized

water and bring volume to 500.0mL Mix thoroughly Autoclave for 15

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

Preparation of Medium: Aseptically combine 500.0mL solution A

and 500.0mL solution B The final pH should be 9.5 Pour into Petri

dishes or distribute into sterile tubes

Use: For the cultivation and maintenance of Aquisalimonas spp.

Aquisalimonas Medium

Solution A 500.0mL

Solution B 500.0mL

pH 9.5 ± 0.2 at 25°C

Solution A:

Glucose 10.0g Peptone 5.0g Yeast extract 5.0g

K2HPO4 1.0g MgCl2·6H2O 0.2g

Preparation of Solution A: Add components to distilled/deionized water and bring volume to 500.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Cool to room temperature

Solution B:

NaCl 80.0g

Na2CO3 20.0g

Preparation of Solution B: Add components to distilled/deionized water and bring volume to 500.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Cool to room temperature

Preparation of Medium: Aseptically combine 500.0mL solution A and 500.0mL solution B The final pH should be 9.5

Use: For the cultivation and maintenance of Aquisalimonas spp.

Arabinose Agar Base with Selective Supplement

Peptone, special 23.0g Agar 15.0g Arabinose 10.0g NaCl 5.0g Corn starch 1.0g Phenol Red 0.1g Selective supplement solution 10.0mL

pH 7.8 ± 0.2 at 25°C

Thallium acetate 0.2g Nalidixic acid 25.0mg

Preparation of Medium: Add components, except selective sup-plement solution, to distilled/deionized water and bring volume to 990.0mL Mix thoroughly Adjust pH to 7.8 Gently heat and bring to boiling Distribute into tubes or flasks Gently heat and bring to boil

Do not autoclave Cool to 50°C Add selective supplement solution Mix thoroughly Pour into sterile Petri dishes

Use: For selective isolation of Enterococcus faecium from feces,

sew-age, and water supplies

ARC51 Medium (DSMZ Medium 1098)

Sea salts, Sigma 35.0g Sodium acetate 1.6g Mercaptoethanesulfonic acid (coenzyme M) 0.14g

NH4Cl 0.1g Yeast extract 0.1g

K2HPO4 0.05g

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142 Archaeoglobus Medium

Resazurin 0.5mg

NaHCO3 solution 10.0mL

Na2S·9H2O solution 10.0mL

Na2S2O3 solution 10.0mL

Wolfe’s mineral elixer 1.0mL

Seven vitamin solution 1.0mL

pH 6.5 ± 0.2 at 25°C

Seven Vitamin Solution:

Pyridoxine hydrochloride 300.0mg

Thiamine-HCl·2H2O 200.0mg

Nicotinic acid 200.0mg

Vitamin B12 100.0mg

Calcium pantothenate 100.0mg

p-Aminobenzoic acid 80.0mg

D(+)-Biotin 20.0mg

Preparation of Seven Vitamin Solution: Add components to

dis-tilled/deionized water and bring volume to 1.0L Sparge with 100% N2

Mix thoroughly Filter sterilize

Wolfe’s Mineral Elixir:

MgSO4·7H2O 30.0g

NaCl 10.0g

MnSO4·2H2O 5.0g

(NH4)2NiSO4·6H2O 2.8g

CoCl2·6H2O 1.8g

ZnSO4·7H2O 1.8g

FeSO4·7H2O 1.0g

CaCl2·2H2O 1.0g

KAl(SO4)2·12H2O 0.18g

CuSO4·5H2O 0.1g

H3BO3 0.1g

Na2MoO4·2H2O 0.1g

Na2SeO4 0.1g

Na2WO4·2H2O 0.1g

Preparation of Wolfe’s Mineral Elixir: Adjust pH of 1.0L of

dis-tilled/deionized water to 1.0 with dilute H2SO4 Add remaining

com-ponents one at a time Mix thoroughly to dissolve

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

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

to room temperature

Na 2 S 2 O 3 Solution :

Na2S2O3·5H2O 2.5g

Preparation of NaHCO 3 Solution: Add Na2S2O3·5H2O to

Sparge with N2 Autoclave for 15 min at 15 psi pressure–121°C Cool

to room temperature

NaHCO 3 Solution :

NaHCO3 2.0g

Preparation of NaHCO 3 Solution: Add NaHCO3 to

distilled/de-ionized water and bring volume to 50.0mL Mix thoroughly Sparge

with N2 Autoclave for 15 min at 15 psi pressure–121°C Cool to room temperature

Preparation of Medium: Add components, except vitamin solu-tion, bicarbonate solusolu-tion, thiosulfate solusolu-tion, and Na2S·9H2O solu-tion, to distilled/deionized water and bring volume to 970.0mL Mix thoroughly Gently heat and bring to boiling Boil for 3 min Cool to room temperature while sparging with 20% CO2 + 80% N2 Dispense into balch tubes or serum bottles under atmosphere of 20% CO2 + 80%

N2 Autoclave for 15 min at 15 psi pressure–121°C Cool to 25°C under

an atmosphere of 20% CO2 + 80% N2 Aseptically and anaerobically add sterile vitamin solution, bicarbonate solution, thiosulfate solution, and Na2S·9H2O solution Adjust final pH to 6.5 After inoculation ad-just overpressure to 1.5 bar with 20% CO2 + 80% N2

Use: For the cultivation and maintenance of Archaeoglobus infectus

Archaeoglobus Medium

NaCl 18.0g NaHCO3 5.0g MgCl2·6H2O 4.0g MgSO4·7H2O 3.45g Sodium L-lactate 1.5g Yeast extract 0.5g KCl 0.34g

NH4Cl 0.25g CaCl2·2H2O 0.14g

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

Na2S·9H2O solution 25.0mL Trace elements solution 10.0mL

pH 6.9 ± 0.2 at 25°C

MgSO4·7H2O 3.0g Nitrilotriacetic acid 1.5g NaCl 1.0g MnSO4·2H2O 0.5g CoSO4·7H2O 0.18g ZnSO4·7H2O 0.18g FeSO4·7H2O 0.1g CaCl2·2H2O 0.1g NiCl2·6H2O 0.025g KAl(SO4)2·l2H2O 0.02g CuSO4·5H2O 0.01g

H3BO3 0.01g

Na2MoO4·2H2O 0.01g

Na2SeO3·5H2O 0.3mg

Na 2 S·9H 2 O Solution:

Na2S·9H2O 1.0g

Preparation of Na 2 S·9H 2 O Solution: Prepare and dispense solu-tion anaerobically under 80% N2 + 20% CO2 Add Na2S·9H2O to dis-tilled/deionized water and bring volume to 50.0mL Mix thoroughly Adjust pH to 7.0 Autoclave for 15 min at 15 psi pressure–121°C

Preparation of Trace Elements Solution: Add nitrilotriacetic acid to approximately 500.0mL of distilled/deionized water Dissolve

by adding KOH and adjust pH to 6.5 Add remaining components

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Archaeoglobus veneficus Medium 143

Bring volume to 1.0L with additional distilled/deionized water Adjust

pH to 7.0 with KOH

Preparation of Medium: Add components, except NaHCO3 and

Na2S·9H2O solution, to distilled/deionized water and bring volume to

1.0L Mix well and heat to boiling for a few minutes Cool rapidly to

room temperature while gassing with 80% N2 + 20% CO2 Add

NaHCO3 and adjust pH to 6.9 Distribute anaerobically under 80% N2

+ 20% CO2 and pressurize sealed containers up to 2 bar pressure

Au-toclave for 15 min at 15 psi pressure–121°C Prior to inoculation of

cultures, add 0.25mL of sterile Na2S·9H2O solution to each tube

con-taining 9.75mL of sterile basal medium

Use: For the cultivation and maintenance of Archaeoglobus fulgidus.

Archaeoglobus profundus Medium

NaCl 18.0g

MgCl2·6H2O 4.0g

MgSO4·7H2O 3.45g

Na2SO4 2.7g

Sodium acetate 1.0g

NaHCO3 1.0g

Yeast extract 0.5g

KCl 0.34g

NH4Cl 0.25g

CaCl2·2H2O 0.14g

K2HPO4 0.14g

Fe(NH4)2(SO4)2·7H2O 2.0mg

Resazurin 1.0mg

Na2S·9H2O solution 25.0mL

pH 6.5 ± 0.2 at 25°C

MgSO4·7H2O 3.0g

Nitrilotriacetic acid 1.5g

NaCl 1.0g

MnSO4·2H2O 0.5g

CoSO4·7H2O 0.18g

ZnSO4·7H2O 0.18g

FeSO4·7H2O 0.1g

CaCl2·2H2O 0.1g

NiCl2·6H2O 0.025g

KAl(SO4)2·l2H2O 0.02g

CuSO4·5H2O 0.01g

H3BO3 0.01g

Na2MoO4·2H2O 0.01g

Na2SeO3·5H2O 0.3mg

Na2S·9H2O 1.0g

Preparation of Trace Elements Solution: Add nitrilotriacetic

acid to approximately 500.0mL of distilled/deionized water Dissolve

by adding KOH and adjust pH to 6.5 Add remaining components

Bring volume to 1.0L with additional distilled/deionized water Adjust

pH to 7.0 with KOH

Preparation of Na 2 S·9H 2 O Solution: Prepare and dispense

solu-tion anaerobically under 80% N2 + 20% CO2 Add Na2S·9H2O to

Adjust pH to 7.0 Autoclave for 15 min at 15 psi pressure–121°C

Preparation of Medium: Add components, except NaHCO3 and

Na2S·9H2O solution, to distilled/deionized water and bring volume to 1.0L Mix well and heat to boiling for a few minutes Cool rapidly to room temperature while gassing with 80% N2 + 20% CO2 Add NaHCO3 and adjust pH to 6.9 Distribute anaerobically under 80% N2 + 20% CO2 and pressurize sealed containers up to 2 bar pressure Autoclave for 15 min at

15 psi pressure–121°C Prior to inoculation of cultures, add 0.25mL of sterile Na2S·9H2O solution to each tube containing 9.75mL of sterile basal medium

Use: For the cultivation and maintenance of Archaeoglobus

profun-dus

Archaeoglobus veneficus Medium

NaCl 18.0g MgCl2·6H2O 7.15g NaHCO3 5.0g KCl 0.33g

NH4Cl 0.25g

K2HPO4·3H2O 0.18g CaCl2·2H2O 0.14g Fe(NH4)2(SO4)2·6H2O 2.0mg Resazurin 0.5mg

Na2SO3 solution 20.0mL

Na2S·9H2O solution 10.0mL Na-acetate solution 4.0mL Trace elements solution 1.0mL

pH 1.0 ± 0.2 at 25°C

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 20.0mL Sparge with N2 Autoclave for 15 min at 15 psi pressure–121°C Cool to 25°C Store an-aerobically

Na 2 SO 3 Solution:

Na2SO3 0.5g

Preparation of Na 2 SO 3 Solution : Add Na2SO3 to distilled/deion-ized water and bring volume to 10.0mL Mix thoroughly Sparge with 100% N2 Filter sterilize

Na-acetate Solution:

Na-acetate 2.5g

Preparation of Na-acetate Solution : Add Na-acetate to distilled/

deionized water and bring volume to 10.0mL Mix thoroughly Sparge with 100% N2 Filter sterilize

NaCl 5.0g MnCl2·4H2O 2.9g (NH4)2Ni(SO4)2 1.0g FeSO4·7H2O 0.5g CoCl2·6H2O 0.5g CaCl2·2H2O 0.5g ZnSO4·7H2 0.5g CuSO4·5H2O 0.05g

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144 Archangium violaceum Medium

H3BO3 0.05g

KAl(SO4)2·12H2O 0.05g

Na2MoO4·4H2O 0.05g

Na2WO4·2H2O 0.05g

Na2SeO3·5H2O 0.05g

Preparation of Trace Elements olution: Add components to

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

Sparge with 100% N2

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2 gas mixture Add components, except Na-acetate

solu-tion, Na2S·9H2O solution, and NaHCO3, to 986.0mL

distilled/deion-ized water Mix thoroughly Gently heat and bring to boiling Boil for

5 min Cool to 25°C while sparging with 80% N2 + 20% CO2 Add the

solid NaHCO3 Equilibrate with the 80% N2 + 20% CO2 gas Adjust

pH to 7.0 with HCl Add 10.0mL Na2S·9H2O solution Distribute into

serum bottles under 80% N2 + 20% CO2 gas; 25.0mL medium in

100mL serum bottles Adjust pH to 1.0 Autoclave for 15 min at 15 psi

pressure–121°C Cool to 25°C Aseptically and anaerobically inject

0.1mL sterile Na-acetate solution and 0.5mL Na2SO3 solution per

25.0mL medium Mix thoroughly After inoculation pressurize to 1 bar

atmosphere with 80% H2 + 20% CO2 gas

Use: For the cultivation of Archaeoglobus veneficus.

Archangium violaceum Medium

Monosodium glutamate 1.0g

L-Leucine 0.5g

L-Tyrosine 0.5g

L-Isoleucine 0.3g

L-Proline 0.25g

MgSO4·7H2O 0.2g

L-Lysine 0.15g

L-Arginine 0.1g

L-Asparagine 0.1g

L-Serine 0.1g

L-Threonine 0.1g

L-Valine 0.1g

L-Alanine 0.05g

L-Glycine 0.05g

L-Histidine 0.05g

L-Methionine 0.05g

Ca3(PO4)2 0.02g

KCl 0.02g

Tris(hydroxymethyl)aminomethane

buffer (0.02m solution, pH 7.5) 1.0L

pH 7.5 ± 0.2 at 25°C

Preparation of Medium: Add solid components to 1.0L of Tris

buffer Mix thoroughly Filter sterilize Aseptically distribute into tubes

or flasks

Use: For the cultivation of Archangium violaceum.

Arcobacter Broth Base with Selective Supplement

Peptone 18.0g

NaCl 5.0g

Yeast extract 1.0g

Selective supplement solution 10.0mL

pH 7.2 ± 0.2 at 25°C

Cefoperazone 16.0mg Amphotericin B 5.0mg

Preparation of Medium: Add components, except selective sup-plement solution, to distilled/deionized water and bring volume to 990.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure– 121°C Cool to 50°C Aseptically add selective supplement solution Mix thoroughly Pour into Petri dishes or aseptically distribute into sterile tubes

Use: For the enrichment and cultivation of Arcobacter spp.

Arcobacter Medium

Agar 15.0g Special peptone 10.0g Beef extract 5.0g NaCl 5.0g Yeast extract 5.0g Sodium glutamate 2.0g Sodium succinate 2.0g MgCl2·H2O 1.0g Horse blood, defibrinated 50.0mL

pH 7.0 ± 0.2 at 25°C

Source: Special peptone is available from Oxoid Unipath

Preparation of Medium: Add components, except horse blood, to distilled/deionized water and bring volume to 950.0mL Mix

thorough-ly Gently heat and bring to boiling Adjust pH to 7.0 Autoclave for 15 min at 15 psi pressure–121°C Cool to 50°–55°C Aseptically add 50.0mL of sterile horse blood Mix thoroughly Pour into sterile Petri dishes or distribute into sterile tubes

Use: For the cultivation and maintenance of Arcobacter species

Arcobacter nitrofigilis Agar

(LMG Medium 86)

Agar 15.0g NaCl 15.0g Lab-Lemco beef extract 10.0g Special peptones, Oxoid 10.0g

pH 7.1 ± 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 Arcobacter nitrofigilis.

Arenavirus Plaquing Medium

Eagle’s basal medium 1.0L Agarose solution 1.0L Fetal calf serum, inactivated 100.0mL

pH 7.0 ± 0.2 at 25°C

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