Handbook of Microbiological Media, Fourth Edition part 129 pdf

0.01g Preparation of Trace Elements Solution SL-6: Add components to distilled/deionized water and bring volume to 1.0L.. Preparation of Medium: Add components, except solution A, so-lut

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Natroniella Medium 1275

Trace Elements Solution SL-6:

Composition per liter:

MnCl2·4H2O 0.5g

H3BO3 0.3g

CoCl2·6H2O 0.2g

ZnSO4·7H2O 0.1g

Na2MoO4·2H2O 0.03g

NiCl2·6H2O 0.02g

CuCl2·2H2O 0.01g

Preparation of Trace Elements Solution SL-6: Add components

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

Preparation of Medium: Add components, except solution A,

so-lution B, and trace elements soso-lution SL-4, to distilled/deionized water

and bring volume to 900.0mL Mix thoroughly Autoclave for 15 min

at 15 psi pressure–121°C Aseptically add 100.0mL of sterile solution

A, 3.0mL of sterile solution B, and 1.0mL of sterile trace elements

so-lution SL-4 Mix thoroughly Aseptically distribute into sterile tubes or

flasks

Use: For the cultivation of Pseudomonas putida.

Natranaerobius Medium

(DSMZ Medium 1095) Composition per liter:

NaCl 100.0g

Yeast extract 10.0g

Tryptone 10.0g

NH4Cl 0.5g

KH2PO4 0.2g

MgCl2·6H2O 0.1g

Sucrose solution 10.0mL

Vitamin solution 10.0mL

Cysteine solution 10.0mL

Bicarbonate solution 10.0mL

Carbonate solution 10.0mL

Trace elements solution SL-10 1.0mL

pH 8.5 ± 0.2 at 25°C

Vitamin Solution:

Pyridoxine-HCl 10.0mg

Thiamine-HCl·2H2O 5.0mg

Riboflavin 5.0mg

Nicotinic acid 5.0mg

D-Ca-pantothenate 5.0mg

p-Aminobenzoic acid 5.0mg

Lipoic acid 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 Sparge

with 80% H2 + 20% CO2 Filter sterilize

MgSO4·7H2O 3.0g

Nitrilotriacetic acid 1.5g

NaCl 1.0g

MnSO4·2H2O 0.5g

CoSO4·7H2O 0.18g

ZnSO4·7H2O 0.18g

CaCl2·2H2O 0.1g FeSO4·7H2O 0.1g NiCl2·6H2O 0.025g KAl(SO4)2·12H2O 0.02g

H3BO3 0.01g

Na2MoO4·4H2O 0.01g CuSO4·5H2O 0.01g

Na2SeO3·5H2O 0.3mg

Preparation of Trace Elements Solution SL-10: Add nitrilotri-acetic acid to 500.0mL of distilled/deionized water Dissolve by adjust-ing pH to 6.5 with KOH Add remainadjust-ing components Add distilled/ deionized water to 1.0L Mix thoroughly Adjust pH to 7.0

Sucrose Solution :

Composition per 10.0mL:

Sucrose 5.0g

Preparation of Sucrose Solution: Add sucrose to distilled/deion-ized water and bring volume to 10.0mL Mix thoroughly Filter steril-ize

Carbonate Solution :

Na2CO3 68.0g

Preparation of Carbonate Solution: Add Na2CO3 to distilled/de-ionized water and bring volume to 10.0mL Mix thoroughly Filter ster-ilize

Bicarbonate Solution :

NaHCO3 38.0g

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

Cysteine Solution:

L-Cysteine-HCl·2H2O 0.07g

Preparation of Cysteine Solution: Add L-Cysteine-HCl·2H2O to

to distilled/deionized water and bring volume to 10.0mL Mix thor-oughly Sparge with 100% N2 Filter sterilize

Preparation of Medium: Add components, except carbonate, bi-carbonate, cysteine, sucrose, and vitamin solutions, to distilled/deion-ized water and bring volume to 950.0mL Mix thoroughly Sparge wtih 100% N2 gas Gently heat and bring to boiling Boil for 1 min Cool to room temperature while sparging with 100% N2 Add cysteine, carbon-ate, and bicarbonate solutions Adjust pH to 8.5 Autoclave for 15 min at

15 psi pressure–121°C Cool to room temperature Aseptically add the vitamins and sucrose solutions Aseptically and anoxically dispense into cuture vessels

Use: For the cultivation of Natranaerobius spp.

Natroniella Medium

(DSMZ Medium 784) Composition per liter:

Na2CO3 68.3g NaCl 15.7g

NH4Cl 1.0g KCl 0.2g

KH2PO4 0.2g Yeast extract 0.2g MgCl2·6H2O 0.1g

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1276 Natroniella Medium

Resazurin 0.5mg

NaHCO3 solution 100.0mL

Na2S·9H2O solution 10.0mL

Ethanol solution 10.0mL

pH 9.7–10.0 at 25°C

Na 2 S·9H 2 O Solution :

Na2S·9H2O 1.0g

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

NaHCO 3 Solution:

NaHCO3 38.3g

Preparation of NaHCO 3 Solution: Add NaHCO3 to

distilled/de-ionized water and bring volume to 100.0mL Mix thoroughly Sparge

with 80% N2 + 20% CO2 Filter sterilize

Ethanol Solution:

Ethanol 5.0mL

Preparation of Ethanol Solution: Add ethanol to

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

100% N2 Filter sterilize

Riboflavin 5.0mg

Lipoic acid 5.0mg

Biotin 2.0mg

Folic acid 2.0mg

Vitamin B12 0.1mg

Composition per liter:

EDTA 0.5g

FeSO4·7H2O 0.2g

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 Ad-just pH to 3.4

Preparation of Medium: Prepare and dispense medium under 100% N2 Add components, except vitamin solution, NaHCO3 solu-tion, ethanol solusolu-tion, and Na2S·9H2O solution, to distilled/deionized water and bring volume to 870.0mL Mix thoroughly Gently heat and bring to boiling Cool to room temperature while sparging with 100%

N2 Autoclave for 15 min at 15 psi pressure–121°C Cool to 25°C Aseptically and anaerobically add 10.0mL sterile vitamin solution, 100.0mL of sterile NaHCO3 solution, 10.0mL sterile ethanol solution, and 10.0mL of sterile Na2S·9H2O solution Mix thoroughly Adjust pH

to 9.7–10.0 Aseptically and anaerobically distribute into sterile tubes

or flasks

Use: For the cultivation of Natroniella acetigena (Acetohalobium sp.).

Natroniella Medium

Na2CO3 68.3g NaCl 15.7g

NH4Cl 1.0g KCl 0.2g

KH2PO4 0.2g Yeast extract 0.2g MgCl2·6H2O 0.1g Resazurin 0.5mg NaHCO3 solution 100.0mL Vitamin solution 10.0mL

Na2S·9H2O solution 10.0mL Ethanol solution 10.0mL Trace elements solution SL-4 1.0mL

pH 9.7–10.0 at 25°C

Na2S·9H2O 1.0g

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

NaHCO3 38.3g

Preparation of NaHCO 3 Solution: Add NaHCO3 to distilled/de-ionized water and bring volume to 100.0mL Mix thoroughly Sparge with 80% N2 + 20% CO2 Filter sterilize

Ethanol Solution:

Ethanol 5.0mL

Preparation of Ethanol Solution: Add ethanol to distilled/deion-ized water and bring volume to 10.0mL Mix thoroughly Sparge with 100% N2 Filter sterilize

Pyridoxine-HCl 10.0mg Thiamine-HCl·2H2O 5.0mg

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Natronincola histidinovorans Medium 1277

Riboflavin 5.0mg

Lipoic acid 5.0mg

Biotin 2.0mg

Folic acid 2.0mg

Vitamin B12 0.1mg

FeCl2·4H2O 1.5g

CoCl2·6H2O 190.0mg

MnCl2··4H2O 100.0mg

ZnCl2· 70.0mg

Na2MoO4·H2O 36.0mg

NiCl2·6H2O 24.0mg

H3BO3 6.0mg

Na2-EDTA 5.2g

CuCl2··2H2O 2.0mg

Adjust pH to 6.0

Preparation of Medium: Prepare and dispense medium under

100% N2 Add components, except vitamin solution, NaHCO3

solu-tion, ethanol, and Na2S·9H2O solution, to distilled/deionized water and

bring volume to 870.0mL Mix thoroughly Gently heat and bring to

boiling Cool to room temperature while sparging with 100% N2

Au-toclave for 15 min at 15 psi pressure–121°C Cool to 25°C Aseptically

and anaerobically add 10.0mL sterile vitamin solution, 100.0mL of

sterile NaHCO3 solution, 10.0mL sterile ethanol solution, and 10.0mL

of sterile Na2S·9H2O solution Mix thoroughly Adjust pH to 9.7–10.0

Aseptically and anaerobically distribute into sterile tubes or flasks

Use: For the cultivation of Natroniella acetigena (Acetohalobium sp.).

Natronincola histidinovorans Medium

NaCl 80.0g

Na2CO3 6.83g

NaHCO3 3.83g

NH4Cl 1.0g

KCl 0.2g

KH2PO4 0.2g

MgCl2·6H2O 0.1g

Resazurin 0.01g

Histidine solution 50.0mL

Na2S·9H2O solution 10.0mL

Yeast extract solution 10.0mL

Trace elements solution 1.0mL

pH 8.9 ± 0.2 at 25°C

Histidine Solution:

Compositionper 50.0mL:

Histidine 5.0g

Preparation of Histidine Solution: Add histidine to

with 100% N2 Autoclave for 15 min at 15 psi pressure–121°C Cool to room temperature

Yeast Extract Solution:

Yeast extract 0.2g

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 under 100% N2 for 15 min at 15 psi pressure–121°C Cool to room temperature

Na2S·9H2O 0.2g

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

Trace Elements Solution:

MgSO4·7H2O 3.0g Nitrilotriacetic acid 1.5g NaCl 1.0g MnSO4·2H2O 0.5g CoSO4·7H2O 0.18g ZnSO4·7H2O 0.18g CaCl2·2H2O 0.1g FeSO4·7H2O 0.1g NiCl2·6H2O 0.025g KAl(SO4)2·12H2O 0.02g

H3BO3 0.01g

Na2MoO4·4H2O 0.01g CuSO4·5H2O 0.01g

Na2SeO3·5H2O 0.3mg

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

to 6.5 with KOH Add remaining components Add distilled/deionized water to 1.0L Mix thoroughly

Pyridoxine-HCl 10.0mg Thiamine-HCl·2H2O 5.0mg Riboflavin 5.0mg Nicotinic acid 5.0mg

Lipoic acid 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 Sparge with 80% H2 + 20% CO2 Filter sterilize

Preparation of Medium: Prepare and dispense medium under 100%

N2 gas atmosphere Add components, except NaHCO3, NH4Cl,

Na2CO3, histidine solution, Na2S·9H2O solution, yeast extract solution, and vitamin solution, to distilled/deionized water and bring volume to 928.0mL Mix thoroughly Gently heat and bring to boiling Boil for 5 min Cool to room temperature while sparging with 100% N2 Add solid NaHCO3, NH4Cl, and Na2CO3 Mix thoroughly Distribute into

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anaer-1278 Natronobacteria Medium

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

Aseptically and anaerobically add per liter of medium 50.0mL histidine

solution, 10.0mL yeast extract solution, 10.0mL Na2S·9H2O solution,

and 2.0mL vitamin solution The final pH should be 8.9

Use: For the cultivation of Natronincola histidinovorans.

Natronobacteria Medium

NaCl 200.0g

Agar 20.0g

Yeast extract 5.0g

Casamino acids 5.0g

KH2PO4 1.0g

KCl 1.0g

NH4Cl 1.0g

Sodium glutamate 1.0g

MgSO4·7H2O 0.24g

CaSO4·2H2O 0.17g

Na2CO3 solution 100.0mL

pH 9.0 ± 0.2 at 25°C

Na 2 CO 3 Solution:

Na2CO3 5.0g

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 50°C

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

Adjust pH to 3.4

Preparation of Medium: Add components, except Na2CO3

solu-tion, to distilled/deionized water and bring volume to 900.0mL Mix

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

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

Na2CO3 solution Mix thoroughly Adjust pH to 9.0, if necessary Pour

into sterile Petri dishes or distribute into sterile tubes

Use: For the cultivation and maintenance of Natronobacterium

grego-ryi, Natronobacterium magadii, Natronobacterium pharaonis, and

Natronococcus occultus.

Natronobacterium Medium

NaCl 250.0g

Agar 20.0g

Casamino acids 15.0g

Trisodium citrate·2H2O 3.0g

Glutamic acid 2.5g

MgSO4·7H2O 2.5g

KCl 2.0g

Na2CO3 solution variable

pH 8.5 ± 0.2 at 25°C

Na2CO3 Solution:

Na2CO3 5.0g

Preparation of Na2CO3 Solution: Add Na2CO3 to distilled/de-ionized water and bring volume to 100.0mL Mix thoroughly Filter sterilize

Preparation of Medium: Add components, except salt and

Na2CO3 solution, to distilled/deionized water and bring volume to 850.0mL Mix thoroughly Gently heat and bring to boiling Add salt Mix thoroughly Bring pH to 7.0 Autoclave for 15 min at 15 psi pres-sure–121°C Cool to 50°–55°C Adjust pH to 8.5 with sterile Na2CO3 solution

Use: For the cultivation and maintenance of Natronobacterium

mag-adii, Natronobacterium pharaonis, and Natronococcus occultus.

Natronobacterium pharaonis Medium

NaCl 250.0g Casamino acids 15.0g Sodium citrate 3.0g Glutamic acid 2.5g MgSO4·7H2O 2.0g KCl 2.0g

pH 8.5 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized wa-ter and bring volume to 1.0L Mix thoroughly Adjust pH to 8.5 Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C

Use: For the cultivation of Natronobacterium pharaonis.

Nautilia Medium

Sulfur 10.0g

NH4Cl 0.33g

KH2PO4 0.33g Resazurin 0.5mg Synthetic seawater, concentrated 500.0mL NaHCO3 solution 50.0mL

Na2S·9H2O solution 20.0mL Sodium formate solution 15.0mL Trace elements solution 10.0mL Vitamin solution 10.0mL Selenite-tungstate solution 1.0mL

pH 6.8 ± 0.2 at 25°C

Synthetic Seawater, Concentrated:

NaCl 55.4g MgSO4·7H2O 14.0g MgCl2·6H2O 11.0g CaCl2·2H2O 1.5g KCl 1.3g NaBr 0.2g

H3BO3 0.06g SrCl2·6H2O 0.03g

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Nautilia Medium 1279

Na3-citrate 20.0mg

KI 0.1mg

Preparation of Synthetic Seawater, Concentrated: Add

com-ponents to distilled/deionized water and bring volume to 500.0mL Mix

thoroughly

MgSO4·7H2O 3.0g

Nitrilotriacetic acid 1.5g

NaCl 1.0g

MnSO4·2H2O 0.5g

CoSO4·7H2O 0.18g

ZnSO4·7H2O 0.18g

CaCl2·2H2O 0.1g

FeSO4·7H2O 0.1g

NiCl2·6H2O 0.025g

KAl(SO4)2·12H2O 0.02g

H3BO3 0.01g

Na2MoO4·4H2O 0.01g

CuSO4·5H2O 0.01g

Na2SeO3·5H2O 0.3mg

Preparation of Trace Elements Solution: Add nitrilotriacetic

acid to 500.0mL of distilled/deionized water Dissolve by adjusting pH

to 6.5 with KOH Add remaining components Add distilled/deionized

water to 1.0L Mix thoroughly

Riboflavin 5.0mg

Lipoic acid 5.0mg

Biotin 2.0mg

Folic acid 2.0mg

Vitamin B12 0.1mg

Sodium Formate Solution:

Na-formate 10.0g

Preparation of Sodium Formate Solution: Add sodium formate

to distilled/deionized water and bring volume to 50.0mL Mix

thor-oughly Sparge with 100% N2 Filter sterilize

Na2S·9H2O 0.6g

to room temperature

NaHCO3 5.0g

Preparation of NaHCO 3 Solution: Add NaHCO3 to distilled/de-ionized water and bring volume to 100.0mL Mix thoroughly Sparge with 80% N2 + 20% CO2 Filter sterilize

Selenite-Tungstate Solution Composition per liter:

NaOH 0.5g

Na2WO4·2H2O 4.0mg

Na2SeO3·5H2O 3.0mg

Preparation of Selenite-Tungstate Solution: Add components

to distilled/deionized water and bring volume to 1.0L Mix thoroughly Sparge with 100% N2 Filter sterilize

Preparation of Sulfur: Sterilize sulfur by steaming for 3 h on each

of 3 successive days

N2 + 20% CO2 gas atmosphere Add components, except sulfur, NaHCO3 solution, sodium formate solution, Na2S·9H2O solution, vita-min solution, selenite-tungstate solution, and trace elements solution,

to distilled/deionized water and bring volume to 894.0mL Mix thor-oughly Adjust pH to 6.8 Sparge with 80% N2 + 20% CO2 Autoclave for 15 min at 15 psi pressure–121°C Aseptically and anaerobically add 10.0g steam sterilized sulfur, 50.0mL NaHCO3 solution, 15.0mL

sodi-um formate solution, 20.0mL Na2S·9H2O solution, 10.0mL vitamin so-lution, 1.0mL selenite-tungstate soso-lution, and 10.0mL trace elements solution Mix thoroughly Adjust pH to 6.8 Aseptically and

anaerobical-ly distribute into sterile tubes or bottles

Use: For the cultivation of Caldithrix abyssi and Nautilia lithotrophica.

Nautilia Medium

NH4Cl 0.33g

KH2PO4 0.33g Resazurin 0.5mg Synthetic seawater, concentrated 500.0mL NaHCO3 solution 50.0mL

Na2S·9H2O solution 20.0mL Trace elements solution 10.0mL Vitamin solution 10.0mL Selenite-tungstate solution 1.0mL

pH 6.8 ± 0.2 at 25°C

Synthetic Seawater, Concentrated:

NaCl 55.4g MgSO4·7H2O 14.0g MgCl2·6H2O 11.0g CaCl2·2H2O 1.5g KCl 1.3g NaBr 0.2g

H3BO3 0.06g SrCl2·6H2O 0.03g

Na3-citrate 20.0mg

KI 0.1mg

MgSO4·7H2O 3.0g Nitrilotriacetic acid 1.5g NaCl 1.0g

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1280 NBA Medium

MnSO4·2H2O 0.5g

CoSO4·7H2O 0.18g

ZnSO4·7H2O 0.18g

CaCl2·2H2O 0.1g

FeSO4·7H2O 0.1g

NiCl2·6H2O 0.025g

KAl(SO4)2·12H2O 0.02g

H3BO3 0.01g

Na2MoO4·4H2O 0.01g

CuSO4·5H2O 0.01g

Na2SeO3·5H2O 0.3mg

Preparation of Trace Elements Solution: Add nitrilotriacetic

acid to 500.0mL of distilled/deionized water Dissolve by adjusting pH

to 6.5 with KOH Add remaining components Add distilled/deionized

water to 1.0L Mix thoroughly

Riboflavin 5.0mg

Lipoic acid 5.0mg

Biotin 2.0mg

Folic acid 2.0mg

Vitamin B12 0.1mg

Na2S·9H2O 0.6g

to room temperature

NaHCO3 5.0g

Preparation of NaHCO 3 Solution: Add NaHCO3 to

with 80% N2 + 20% CO2 Filter sterilize

Selenite-Tungstate Solution

NaOH 0.5g

Na2WO4·2H2O 4.0mg

Na2SeO3·5H2O 3.0mg

Preparation of Selenite-Tungstate Solution: Add components

Sparge with 100% N2 Filter sterilize

Yeast Extract Solution:

Yeast extract 4.0g

Preparation of Yeast Extract Solution: Add yeast extract to

Sparge with 100% N2 Autoclave under 100% N2 for 15 min at 15 psi pressure–121°C Cool to room temperature

N2 + 20% CO2 gas atmosphere Add components, except NaHCO3 solu-tion, yeast extract solusolu-tion, Na2S·9H2O solution, vitamin solution, sel-enite-tungstate solution, and trace elements solution, to distilled/ deionized water and bring volume to 894.0mL Mix thoroughly Adjust

pH to 6.8 Sparge with 80% N2 + 20% CO2 Autoclave for 15 min at 15 psi pressure–121°C Aseptically and anaerobically add 50.0mL NaHCO3 solution, 15.0mL yeast extract solution, 20.0mL Na2S·9H2O solution, 10.0mL vitamin solution, 1.0mL selenite-tungstate solution, and 10.0mL trace elements solution Mix thoroughly Adjust pH to 6.8 Aseptically and anaerobically distribute into sterile tubes or bottles

Use: For the cultivation of Caldithrix abyssi DSM 13497.

NBA Medium Composition per liter:

Pancreatic digest of gelatin 5.0g Casamino acids 5.0g Beef extract 3.0g Yeast extract 1.0g

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 Distribute into tubes

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

Use: For the cultivation of Bdellovibrio species.

NBY Medium (Nutrient Broth Yeast Extract Medium) Composition per liter:

Nutrient broth, dehydrated 8.0g Yeast extract 2.0g

K2HPO4 2.0g

KH2PO4 0.5g Glucose solution 50.0mL MgSO4·7H2O (1M solution) 1.0mL

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 Filter steril-ize

Preparation of Medium: Add components, except glucose solu-tion, to distilled/deionized water and bring volume to 950.0mL Mix thoroughly Gently heat and bring to boiling Autoclave for 15 min at

15 psi pressure–121°C Cool to 45°–50°C Aseptically add sterile glu-cose solution Mix thoroughly Pour into sterile Petri dishes or distrib-ute into sterile tubes

Use: For the cultivation and maintenance of Curtobacterium

flaccum-faciens and Pseudomonas syringae.

NBY Medium (Nutrient Broth Yeast Extract Medium)

(ATCC Medium 763) Composition per liter:

Agar 15.0g Nutrient broth 8.0g

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Nelson Culture Medium for Naegleria 1281

Yeast extract 2.0g

K2HPO4 2.0g

KH2PO4 0.5g

Glucose solution 50.0mL

MgSO4 solution 50.0mL

D-Glucose 5.0g

Preparation of Glucose Solution: Add glucose to

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

steril-ize

MgSO 4 Solution:

MgSO4·7H2O 0.25g

Preparation of MgSO 4 Solution: Add the solid MgSO4·7H2O to

distilled/deionized water and bring volume to 50.0mL Mix

thorough-ly Autoclave for 15 min at 15 psi pressure–121°C Cool to 50°C

Preparation of Medium: Add components, except glucose

solu-tion and MgSO4 solution, to distilled/deionized water and bring

vol-ume to 900.0mL Mix thoroughly Gently heat and bring to boiling

Autoclave for 25 min at 15 psi pressure–121°C Cool to 45°–50°C

Aseptically add sterile glucose solution and MgSO4 solution Mix

thor-oughly Pour into sterile Petri dishes or distribute into sterile tubes

Use: For the cultivation and maintenance of Bacillus sphaericus.

Neisseria Medium

Biosate 10.0g

Polypeptone™ 10.0g

NaCl 5.0g

Myosate 3.0g

Agar 1.5g

Phenol Red 0.017g

Carbohydrate solution 50.0mL

pH 7.4–7.6 at 25°C

Carbohydrate Solution:

Carbohydrate 10.0g

Preparation of Carbohydrate Solution: Add glucose, sucrose,

or maltose to distilled/deionized water and bring volume to 50.0mL

Mix thoroughly Filter sterilize

Preparation of Medium: Add components, except carbohydrate

solution, to distilled/deionized water and bring volume to 950.0mL

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

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

carbohydrate solution Mix thoroughly Pour into sterile Petri dishes or

distribute into sterile tubes

Use: For the cultivation of Neisseria species.

Neisseria meningitidis Medium

Beef infusion 300.0g

Acid hydrolysate of casein 17.5g

Agar 17.0g

Starch 1.5g

Antibiotic solution 10.0mL

pH 7.4 ± 0.2 at 25°C

Antibiotic Solution:

Vancomycin 3.0mg Colistin 7.5mg Nystatin 12,500U

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

Preparation of Medium: Add components, except antibiotic solu-tion, to distilled/deionized water and bring volume to 990.0mL Mix thoroughly Gently heat and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C Aseptically add sterile antibiotic solution Mix thoroughly Pour into sterile Petri dishes or distribute into sterile tubes

Use: For the selective isolation and cultivation of Neisseria

meningit-idis.

Nelson Culture Medium for Naegleria

Glucose 0.17g Panmede 0.17g

Na2HPO4 14.2mg

KH2PO4 13.6mg NaCl 12.0mg CaCl2·2H2O 0.4mg MgSO4·7H2O 0.4mg Bovine serum, heat-inactivated fetal 10.0mL

Source: Panmede is available from Paines and Byrne Ltd., Greenford, England, and Harrisons and Crosfield, Inc., Bronxville, NY

Preparation of Medium: Add components, except bovine serum,

to distilled/deionized water and bring volume to 90.0mL Mix thor-oughly Autoclave for 15 min at 15 psi pressure–121°C Cool to 25°C Aseptically add 10.0mL of sterile, heat-inactivated fetal bovine serum Mix thoroughly Aseptically distribute into sterile tubes or flasks Use immediately

Use: For the cultivation of Naegleria fowleri and Paratetramitus

jugo-sus.

Nelson Culture Medium for Naegleria

Glucose 0.17g Liver infusion 0.17g

Na2HPO4 14.2mg

KH2PO4 13.6mg NaCl 12.0mg CaCl2·2H2O 0.4mg MgSO4·7H2O 0.4mg Bovine serum, heat-inactivated fetal 10.0mL

Preparation of Medium: Add components, except bovine serum,

to distilled/deionized water and bring volume to 90.0mL Mix thor-oughly Autoclave for 15 min at 15 psi pressure–121°C Cool to 25°C Aseptically add 10.0mL of sterile, heat-inactivated fetal bovine serum Mix thoroughly Aseptically distribute into sterile tubes or flasks Use immediately

Use: For the cultivation of Naegleria fowleri and Paratetramitus

jugo-sus.

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1282 Nelson Medium for Naegleria fowleri

Nelson Medium for Naegleria fowleri

Glucose 1.0g

Ox liver digest 1.0g

Page’s amoeba saline 1.0L

Fetal calf serum, inactivated 20.0mL

Page’s Amoeba Saline:

Na2HPO4 0.142g

KH2PO4 0.136g

NaCl 0.12g

MgSO4·7H2O 4.0mg

CaCl2·2H2O 4.0mg

Preparation of Page’s Amoeba Saline: Add components to

Preparation of Medium: Add the glucose and ox liver digest to

1.0L of Page’s amoeba saline Mix thoroughly Distribute into

screw-capped tubes in 10.0mL volumes Autoclave for 15 min at 15 psi

pres-sure–121°C Cool to 25°C Aseptically add 0.2mL of sterile fetal calf

serum to each tube Mix thoroughly

Use: For the cultivation of Naegleria fowleri.

Neocallimastix Medium

(ANO2 Fungus II) Composition per liter:

NaHCO3 6.6g

Cellobiose 2.0g

Glucose 2.0g

Maltose 2.0g

Starch 2.0g

PIPES

(piperazine-N,N´-bis[2-ethanesulfonic acid]) buffer 1.0g

Trypticase™ peptone 1.0g

Yeast extract 1.0g

L-Cysteine·HCl·H2O 0.5g

KH2PO4 0.5g

MgCl2·6H2O 0.4g

NaCl 0.4g

NH4Cl 0.4g

Na2S·9H2O 0.1g

CaCl2·2H2O 50.0mg

Resazurin 1.0mg

Rumen fluid 100.0mL

Wolfe’s vitamin solution 10.0mL

pH 6.9 ± 0.2 at 25°C

Wolfe’s Vitamin Solution:

Pyridoxine·HCl 10.0mg

Lipoic acid 5.0mg

Riboflavin 5.0mg

Thiamine·HCl 5.0mg

Calcium DL-pantothenate 5.0mg

Biotin 2.0mg

Folic acid 2.0mg

Vitamin B12 0.1mg

Preparation of Wolfe’s Vitamin Solution: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly

FeCl2·4H2O 1.5g CoCl2·6H2O 190.0mg MnCl2·4H2O 100.0mg ZnCl2 70.0mg

Na2MoO4·2H2O 36.0mg NiCl2·6H2O 24.0mg

H3BO3 6.0mg CuCl2·2H2O 2.0mg HCl (25% solution) 10.0mL

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 components Mix thor-oughly

Preparation of Medium: Prepare and dispense medium under 100% CO2 Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Gently heat and bring to boiling Con-tinue boiling for 3 min Cool to room temperature while sparging with 100% CO2 Anaerobically distribute into tubes Autoclave for 15 min

at 15 psi pressure–121°C Adjust pH to 6.9

Use: For the cultivation of Neocallimastix frontalis.

Neomycin Agar Composition per liter:

Agar 15.0g Peptone 6.0g Pancreatic digest of casein 4.0g Yeast extract 3.0g Beef extract 1.5g Glucose 1.0g Neomycin solution 10.0mL

pH 7.0 ± 0.2 at 25°C

Neomycin Solution:

Neomycin sulfate 1.0g

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

Preparation of Medium: Add components, except neomycin 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 neo-mycin solution Mix thoroughly Pour into sterile Petri dishes or dis-tribute into sterile tubes

Use: For the cultivation and maintenance of Micrococcus luteus.

Neomycin Agar, Modified Composition per liter:

Agar 15.0g Peptone 6.0g Pancreatic digest of casein 4.0g Yeast extract 3.0g Beef extract 1.5g Glucose 1.0g

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Neomycin Medium No 1 1283

Methanol 20.0mL

Neomycin solution 10.0mL

pH 7.0 ± 0.2 at 25°C

Preparation of Neomycin Solution: Add neomycin sulfate to

Filter sterilize

Preparation of Medium: Add components, except methanol and

neomycin solution, to distilled/deionized water and bring volume to

970.0mL Mix thoroughly Gently heat and bring to boiling Autoclave

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

methanol To cooled, sterile basal medium, aseptically add sterile

methanol and sterile neomycin solution Mix thoroughly Pour into

sterile Petri dishes or distribute into sterile tubes

Use: For the cultivation and maintenance of Bordetella

bronchisep-tica.

Neomycin Assay Agar

See: Antibiotic Medium 11

Neomycin Blood Agar Composition per liter:

Pancreatic digest of casein 14.5g

Agar 14.0g

Papaic digest of soybean meal 5.0g

NaCl 5.0g

Growth factors 1.5g

Sheep blood, defibrinated 50.0mL

Neomycin solution 10.0mL

pH 7.3 ± 0.2 at 25°C

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

Di-agnostic Systems

Preparation of Neomycin Solution: Add neomycin sulfate to

Filter sterilize

Preparation of Medium: Add components, except sheep blood and

neomycin solution, to distilled/deionized water and bring volume to

940.0mL 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 sheep blood and sterile neomycin solution Mix thoroughly

Pour into sterile Petri dishes or distribute into sterile tubes

Use: For the isolation and cultivation of group A streptococci

(Strep-tococcus pyogenes) and group B streptococci (Strep(Strep-tococcus

agalac-tiae) from throat cultures and other clinical specimens.

Neomycin Luria Agar Composition per liter:

Agar 15.0g

Pancreatic digest of casein 10.0g

Yeast extract 5.0g NaCl 0.5g Glucose solution 20.0mL Neomycin solution 10.0mL

pH 7.0 ± 0.1 at 25°C

Glucose 10.0g

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

Neomycin sulfate 12.0mg

Preparation of Medium: Add components, except glucose solution and neomycin solution, to distilled/deionized water and bring volume to 970.0mL Mix thoroughly Gently heat and bring to boiling Autoclave for

15 min at 15 psi pressure–121°C Aseptically add 20.0mL of sterile glu-cose solution and 10.0mL of sterile neomycin solution Mix thoroughly Pour into sterile Petri dishes or distribute into sterile tubes

Use: For the cultivation of Escherichia coli.

Neomycin Medium No 1 Composition per liter:

Agar 15.0g Peptone 5.0g NaCl 5.0g Yeast extract 2.0g Beef extract 1.0g Sucrose solution 20.0mL Neomycin solution 10.0mL

pH 7.0 ± 0.1 at 25°C

Sucrose Solution:

Sucrose 2.5g

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

Neomycin sulfate 500.0mg

Preparation of Medium: Add components, except sucrose solution and neomycin solution, to distilled/deionized water and bring volume to 970.0mL Mix thoroughly Gently heat and bring to boiling Autoclave for

15 min at 15 psi pressure–121°C Aseptically add 20.0mL of sterile su-crose solution and 10.0mL of sterile neomycin solution Mix thoroughly Pour into sterile Petri dishes or distribute into sterile tubes

Use: For the cultivation of Micrococcus luteus and Pseudomonas

aerug-inosa.

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1284 Neopeptone Glucose Agar

Neopeptone Glucose Agar

Agar 20.0g

Glucose 10.0g

Neopeptone 5.0g

pH 6.5 ± 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 Distribute into tubes or flasks Autoclave for 15 min at 15

psi pressure–121°C Adjust pH to 6.5 Pour into sterile Petri dishes or

leave in tubes

Use: For the maintenance of stock cultures of a variety of

microorgan-isms

Neopeptone Glucose Rose Bengal Aureomycin® Agar

Agar 20.0g

Neopeptone 5.0g

Glucose 1.0g

Tetracycline solution 5.0mL

Rose Bengal solution 3.5mL

pH 6.5 ± 0.2 at 25°C

Tetracycline Solution:

Tetracycline 1.0g

Preparation of Tetracycline Solution: Add tetracycline to

Filter sterilize

Rose Bengal Solution:

Rose Bengal 1.0g

Preparation of Rose Bengal Solution: Add Rose Bengal to

Filter sterilize

Preparation of Medium: Add components, except tetracycline

so-lution, to distilled/deionized water and bring volume to 995.0mL 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 5.0mL of

sterile tetracycline solution Mix thoroughly Pour into sterile Petri

dishes or distribute into sterile tubes

Use: For the isolation and cultivation of a wide variety of fungal

spe-cies

Neopeptone Infusion Agar

Beef heart, infusion from 500.0g

Neopeptone 20.0g

Agar 20.0g

NaCl 5.0g

Sheep blood, defibrinated 50.0mL

pH 7.4 ± 0.2 at 25°C

Preparation of Medium: Add components, except sheep blood, to

distilled/deionized water and bring volume to 950.0mL Mix

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

pressure–121°C Cool to 45°–50°C Aseptically add sterile sheep

blood Mix thoroughly Pour into sterile Petri dishes or distribute into

sterile tubes

Use: For the cultivation of a wide variety of fastidious microorgan-isms

Nesterenkonia Modified Medium

(DSMZ Medium 993a) Composition per liter:

MgCl2·6H2O 100.0g Agar 20.0g Glycerol 10.0g Yeast extract 5.0g

L-Asparagine, anhydrous 1.0g

K2HPO4, anhydrous 1.0g Trace elements solution 1.0mL

pH 7.2 ± 0.2 at 25°C

FeSO4·7H2O 0.1g MnCl2·4H2O 0.1g ZnSO4·7H2O 0.1g

Preparation of Trace Elements Solution: Add components to distilled/deionized water and bring volume to 100.0mL Mix

thorough-ly

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.2 Distribute into tubes or flasks Gently heat while stirring and bring to boiling Mix thoroughly Autoclave for 15 min at 15 psi pressure– 121°C Pour into Petri dishes or leave in tubes

Use: For the cultivation of Nesterenkonia spp.

Neurospora Culture Agar

Maltose 40.0g Agar 15.0g Proteose peptone No 3 5.0g Yeast extract 5.0g

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 while stirring and bring to boiling Distribute into tubes in 8.0mL volumes Autoclave for 15 min at 15 psi pressure–121°C Allow tubes to cool in

a slanted position

Use: For the cultivation of Neurospora intermedia used in the

micro-biological assay of pyridoxine Also used for the cultivation of other fungi

Neurospora Medium

Agar 15.0g Glucose 5.0g Malt syrup, spray dried 5.0g Sucrose 5.0g Yeast extract 2.5g Vitamin solution 10.0mL Casein, hydrolyzed 5.0mL

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