Handbook of Microbiological Media, Fourth Edition part 104 pps

6.8g Preparation of Sodium Formate Solution: Add sodium formate to distilled/deionized water and bring volume to 50.0mL.. 3.0mg Preparation of Selenite-Tungstate Solution: Add components

Martin-Lewis Agar, Enriched 1025

KCl 2.0g

Glucose 1.0g

CaCl2 0.36g

NaHCO3 0.06g

NaBr 0.026g

pH 7.2 ± 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 and maintenance of Marinococcus albus.

Marinomonas vaga Medium See: Nutrient Agar with 3% NaCl

Marinomonas vaga Medium

(DSMZ Medium 617) Compositionper liter:

NaCl 30.0g

Agar 15.0g

Beef extract 10.0g

Peptone 10.0g

pH 7.1 ± 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

Dis-tribute 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 Marinomonas

commu-nis=Alteromonas communis and Marinomonas vaga=Alteromonas

vaga.

Martin-Lewis Agar Compositionper liter:

Agar 12.0g

Hemoglobin 10.0g

Pancreatic digest of casein 7.5g

Selected meat peptone 7.5g

NaCl 5.0g

K2HPO4 4.0g

Cornstarch 1.0g

KH2PO4 1.0g

Supplement solution 10.0mL

VCAT inhibitor 10.0mL

pH 7.2 ± 0.22 at 25°C

Source: Martin-Lewis agar is available as a prepared medium from

BD Diagnostic Systems

Supplement Solution:

Compositionper liter:

Glucose 100.0g

L-Cysteine·HCl 25.9g

L-Glutamine 10.0g

L-Cystine 1.1g

Adenine 1.0g

Nicotinamide adenine dinucleotide 0.25g

Vitamin B12 0.1g

Thiamine pyrophosphate 0.1g

Guanine·HCl 0.03g

Fe(NO3)3·6H2O 0.02g

p-Aminobenzoic acid 0.013g

Thiamine·HCl 3.0mg

Source: The supplement solution IsoVitaleX® enrichment is available from BD Diagnostic Systems This enrichment may be replaced by supplement VX from BD Diagnostic Systems

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

VCAT Inhibitor:

Compositionper 10.0mL:

Colistin 7.5mg Trimethoprim lactate 5.0mg Vancomycin 4.0mg Anisomycin 0.02g

Preparation of VCAT Inhibitor: Add components to distilled/de-ionized water and bring volume to 10.0mL Mix thoroughly Filter ster-ilize

Preparation of Medium: Add components, except supplement so-lution and VCAT inhibitor, to distilled/deionized water and bring vol-ume to 980.0mL Gently heat while stirring and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C Aseptically add sterile supplement solution and sterile VCAT inhibitor Mix thoroughly Pour into sterile Petri dishes

Use: For the isolation and cultivation of pathogenic Neisseria from

specimens containing mixed flora of bacteria and fungi

Martin-Lewis Agar, Enriched Compositionper liter:

Agar 12.0g Pancreatic digest of casein 7.5g Selected meat peptone 7.5g NaCl 5.0g

K2HPO4 4.0g Cornstarch 1.0g

KH2PO4 1.0g

Sarcina lutea suspension 20.0mL

Horse serum, inactivated 20.0mL Supplement solution 10.0mL PCAT inhibitor 10.0mL

pH 7.2 ± 0.22 at 25°C

Source: The supplement solution (IsoVitaleX® enrichment) is avail-able from BD Diagnostic Systems This enrichment may be replaced

by supplement VX from BD Diagnostic Systems

Sarcina lutea Suspension:

Compositionper 20.0mL:

Sarcina lutea FDA 1001 106–107 cells

Preparation of Sarcina lutea Suspension: Aseptically wash the

growth of 24-hr cultures of Sarcina lutea FDA 1001 cells from

Thayer-Martin plates with sterile soybean casein digest broth Standardize the suspension by adding additional sterile tryptic soy broth to yield 40% light transmission at 530nm wavelength

Soybean Casein Digest Broth:

Compositionper liter:

Pancreatic digest of casein 17.0g NaCl 5.0g Papaic digest of soybean meal 3.0g

K2HPO4 2.5g Glucose 2.5g

pH 7.3 ± 0.2 at 25°C

Preparation of Soybean Casein Digest Broth: 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

Supplement Solution:

Compositionper liter:

Glucose 100.0g

L-Cysteine·HCl 25.9g

L-Glutamine 10.0g

L-Cystine 1.1g

Adenine 1.0g

Nicotinamide adenine dinucleotide 0.25g

Vitamin B12 0.1g

Thiamine pyrophosphate 0.1g

Guanine·HCl 0.03g

Fe(NO3)3·6H2O 0.02g

p-Aminobenzoic acid 0.013g

Thiamine·HCl 3.0mg

Preparation of Supplement Solution: Add components to

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

sterilize

PCAT Inhibitor:

Compositionper 10.0mL:

Anisomycin 0.02g

Colistin 7.5mg

Trimethoprim lactate 5.0mg

Penicillin G 25,000U

Preparation of PCAT Inhibitor: Add components to

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

ster-ilize

Preparation of Medium: Add components—except Sarcina lutea

suspension, horse serum, supplement solution, and PCAT inhibitor—to

distilled/deionized water and bring volume to 940.0mL Gently heat

while stirring and bring to boiling Autoclave for 15 min at 15 psi

pres-sure–121°C Cool to 45°–50°C Aseptically add 20.0mL of sterile

Sarci-na lutea suspension, 20.0mL of sterile horse serum, 10.0mL of

supplement solution, and 10.0mL of sterile PCAT inhibitor Mix

thor-oughly Pour into sterile Petri dishes

Use: For the isolation and cultivation of pathogenic Neisseria,

espe-cially penicillinase-producing strains, from specimens containing

mixed flora of bacteria and fungi

Mating Agar Compositionper liter:

Agar 40.0g

Sucrose 10.0g

Xylose 2.0g

KH2PO4 1.0g

MgSO4 0.5g

Yeast extract 0.5g

CaCl2 0.1g

NaCl 0.1g

Biotin 5.0μg

pH 5.7 ± 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 Adjust pH to 5.7 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 Filobasidiella neofor-mans.

Maximum Recovery Diluent Compositionper liter:

NaCl 8.5g Peptone 1.0g

pH 7.0 ± 0.2 at 25°C

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

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: This diluent is a physiologically isotonic and protective medium for maximal recovery of microorganisms from a variety of sources

M-Azide HiVeg Broth Base with Triphenyltetrazolium Chloride Compositionper liter:

Saccharose 100.0g Plant hydrolysate No 1 40.0g Yeast extract 10.0g

K2HPO4 4.0g Glucose 2.0g NaN3 0.4g Triphenyltetrazolium chloride solution 5.0mL

pH 7.2 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Hi-Media

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

Triphenyltetrazolium Choride Solution:

Triphenyltetrazolium chloride 0.1g

Preparation of Triphenyltetrazolium Choride Solution: Add triphenyltetrazolium chloride to distilled/deionized water and bring volume to 5.0mL Mix thoroughly Filter sterilize

Preparation of Medium: Add components, except triphenyltetrazo-lium chloride solution, 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 Cool

to 45°–50°C Aseptically add 5.0mL triphenyltetrazolium chloride so-lution Mix thoroughly Aseptically distribute into tubes

Use: For the detection and enrichment of fecal streptococci in water and sewage by the membrane filtration method

MB Medium (DSMZ Medium 924) Compositionper liter:

NaCl 10.0g NaHCO3 4.0g Yeast extract 2.0g Trypticase™ 2.0g

NH4Cl 1.0g MgCl2·6H2O 1.0g KCl 0.5g CaCl2·2H2O 0.4g

MB Medium 1027

K2HPO4 0.4g

Resazurin 0.5mg

Sodium formate solution 50.0mL

Na2S·9H2O solution 10.0mL

Cysteine-HCl·H2O solution 10.0mL

Vitamin solution 10.0mL

Trace elements solution 10.0mL

Selenite-tungstate solution 1.0mL

pH 7.2 ± 0.2 at 25°C

Cysteine Solution:

Compositionper 10.0mL:

L-Cysteine·HCl·H2O 0.25g

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

distilled/deionized water and bring volume to 10.0mL Mix

thorough-ly Sparge with 100% N2 Autoclave for 15 min at 15 psi pressure–

121°C

Sodium Formate Solution:

Compositionper 50.0mL:

Na-formate 6.8g

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

Selenite-Tungstate Solution

Compositionper 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

Na 2 S·9H 2 O Solution:

Composition per 10.0mL:

Na2S·9H2O 0.25g

Preparation of Na 2 S·9H 2 O Solution: Add Na2S·9H2O to

dis-tilled/deionized water and bring volume to 10.0mL Sparge with N2

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

an-aerobically

Trace Elements Solution:

Compositionper liter:

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

Vitamin Solution:

Compositionper liter:

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

Preparation of Medium: Prepare and dispense medium under an oxygen-free 80% N2 + 20% CO2 gas mixture Add components, except sodium formate solution, NaHCO3, cysteine solution, and Na2S·9H2O solution, to distilled/deionized water and bring volume to 920.0mL Mix thoroughly Gently heat and bring to boiling Boil for 3 min Cool

to 25°C while sparging with 80% N2 + 20% CO2 Add solid NaHCO3 Mix thoroughly Adjust pH to 6.8–7.0 Distribute into tubes or bottles Autoclave for 15 min at 15 psi pressure–121°C Aseptically and anaer-obically add per liter 50.0mL sterile sodium formate solution, 10.0mL

of sterile cysteine solution, and 10.0mL of sterile Na2S·9H2O solution Mix thoroughly The final pH should be 7.2

Use: For the cultivation of Methanocalculus taiwanensis, Methanococ-cus voltae (MethanococMethanococ-cus voltaei), and Methanofollis aquaemaris.

MB Medium (DSMZ Medium 924) Compositionper liter:

NaCl 5.0g NaHCO3 4.0g Yeast extract 2.0g Trypticase™ 2.0g

NH4Cl 1.0g MgCl2·6H2O 1.0g KCl 0.5g CaCl2·2H2O 0.4g

K2HPO4 0.4g Resazurin 0.5mg Sodium formate solution 50.0mL

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

pH 6.5 ± 0.2 at 25°C

Cysteine Solution:

Compositionper 10.0mL:

L-Cysteine·HCl·H2O 0.25g

Preparation of Cysteine Solution: Add L-cysteine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL Mix

thorough-ly Sparge with 100% N2 Autoclave for 15 min at 15 psi pressure– 121°C

Sodium Formate Solution:

Compositionper 50.0mL:

Na-formate 6.8g

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

Selenite-Tungstate Solution

Compositionper 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

Na 2 S·9H 2 O Solution:

Composition per 10.0mL:

Na2S·9H2O 0.25g

Preparation of Na 2 S·9H 2 O Solution: Add Na2S·9H2O to

dis-tilled/deionized water and bring volume to 10.0mL Sparge with N2

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

an-aerobically

Trace Elements Solution:

Compositionper liter:

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

Vitamin Solution:

Compositionper liter:

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

Preparation of Medium: Prepare and dispense medium under an

oxygen-free 80% N2 + 20% CO2 gas mixture Add components, except

sodium formate solution, NaHCO3, cysteine solution, and Na2S·9H2O

solution, to distilled/deionized water and bring volume to 920.0mL

Mix thoroughly Gently heat and bring to boiling Boil for 3 min Cool

to 25°C while sparging with 80% N2 + 20% CO2 Add solid NaHCO3 Mix thoroughly Adjust pH to 6.8–7.0 Distribute into tubes or bottles Autoclave for 15 min at 15 psi pressure–121°C Aseptically and anaer-obically add per liter 50.0mL sterile sodium formate solution, 10.0mL

of sterile cysteine solution, and 10.0mL of sterile Na2S·9H2O solution Mix thoroughly The final pH should be 6.5

Use: For the cultivation of Methanofollis aquaemaris DSM 14661.

MB Medium (DSMZ Medium 924) Compositionper liter:

NaCl 10.0g NaHCO3 4.0g Yeast extract 2.0g Trypticase™ 2.0g

NH4Cl 1.0g MgCl2·6H2O 1.0g KCl 0.5g CaCl2·2H2O 0.4g

K2HPO4 0.4g Resazurin 0.5mg Sodium acetate solution 50.0mL Sodium formate solution 50.0mL

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

pH 7.2 ± 0.2 at 25°C

Cysteine Solution:

Compositionper 10.0mL:

L-Cysteine·HCl·H2O 0.25g

Preparation of Cysteine Solution: Add L-cysteine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL Mix

thorough-ly Sparge with 100% N2 Autoclave for 15 min at 15 psi pressure– 121°C

Sodium Formate Solution:

Compositionper 50.0mL:

Na-formate 6.8g

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

Sodium Acetate Solution:

Compositionper 50.0mL:

Na-acetate 1.6g

Preparation of Sodium Acetate Solution: Add sodium acetate to distilled/deionized water and bring volume to 50.0mL Mix

thorough-ly Sparge with 100% N2 Filter sterilize

Selenite-Tungstate Solution Compositionper 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

MB Medium 1029

Na 2 S·9H 2 O Solution:

Composition per 10.0mL:

Na2S·9H2O 0.25g

Preparation of Na 2 S·9H 2 O Solution: Add Na2S·9H2O to

dis-tilled/deionized water and bring volume to 10.0mL Sparge with N2

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

an-aerobically

Trace Elements Solution:

Compositionper liter:

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

Vitamin Solution:

Compositionper liter:

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

Preparation of Medium: Prepare and dispense medium under an

oxygen-free 80% N2 + 20% CO2 gas mixture Add components, except

sodium acetate solution, sodium formate solution, NaHCO3, cysteine

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

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

boiling Boil for 3 min Cool to 25°C while sparging with 80% N2 +

20% CO2 Add solid NaHCO3 Mix thoroughly Adjust pH to 6.8–7.0

Distribute into tubes or bottles Autoclave for 15 min at 15 psi

pres-sure–121°C Aseptically and anaerobically add per liter 50.0mL sterile

sodium acetate solution, 50.0mL sterile sodium formate solution,

10.0mL of sterile cysteine solution, and 10.0mL of sterile Na2S·9H2O

solution Mix thoroughly The final pH should be 7.2

Use: For the cultivation of Methanocalculus taiwanensis DSM 14648.

MB Medium (DSMZ Medium 924) Compositionper liter:

NaCl 5.0g NaHCO3 4.0g Yeast extract 2.0g Trypticase™ 2.0g

NH4Cl 1.0g MgCl2·6H2O 1.0g KCl 0.5g CaCl2·2H2O 0.4g

K2HPO4 0.4g Resazurin 0.5mg Sodium acetate solution 50.0mL Sodium formate solution 50.0mL

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

pH 7.2 ± 0.2 at 25°C

Cysteine Solution:

Compositionper 10.0mL:

L-Cysteine·HCl·H2O 0.25g

Preparation of Cysteine Solution: Add L-cysteine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL Mix

thorough-ly Sparge with 100% N2 Autoclave for 15 min at 15 psi pressure– 121°C

Sodium Formate Solution:

Compositionper 50.0mL:

Na-formate 6.8g

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

Sodium Acetate Solution:

Compositionper 50.0mL:

Na-acetate 1.6g

Preparation of Sodium Acetate Solution: Add sodium acetate to distilled/deionized water and bring volume to 50.0mL Mix

thorough-ly Sparge with 100% N2 Filter sterilize

Selenite-Tungstate Solution Compositionper 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

Na 2 S·9H 2 O Solution:

Composition per 10.0mL:

Na2S·9H2O 0.25g

Preparation of Na 2 S·9H 2 O Solution: Add Na2S·9H2O to dis-tilled/deionized water and bring volume to 10.0mL Sparge with N2 Autoclave for 15 min at 15 psi pressure–121°C Cool to 25°C Store an-aerobically

Trace Elements Solution:

Compositionper liter:

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

Vitamin Solution:

Compositionper liter:

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

Preparation of Medium: Prepare and dispense medium under an

oxygen-free 80% N2 + 20% CO2 gas mixture Add components, except

sodium acetate solution, sodium formate solution, NaHCO3, cysteine

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

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

boiling Boil for 3 min Cool to 25°C while sparging with 80% N2 +

20% CO2 Add solid NaHCO3 Mix thoroughly Adjust pH to 6.8–7.0

Distribute into tubes or bottles Autoclave for 15 min at 15 psi

pres-sure–121°C Aseptically and anaerobically add per liter 50.0mL sterile

sodium acetate solution, 50.0mL sterile sodium formate solution,

10.0mL of sterile cysteine solution, and 10.0mL of sterile Na2S·9H2O

solution Mix thoroughly The final pH should be 7.2

Use: For the cultivation of Methanocalculus taiwanensis DSM 14663.

M-BCG Yeast and Mold Agar

Composition per liter:

Glucose 50.0g

Agar 15.0g

Biopeptone 10.0g

Yeast extract 9.0g

MgSO4·7H2O 2.1g

KH2PO4 2.0g

Diastase 0.05g

Thiamine hydrochloride 0.05g Bromcresol Green 0.026g

pH 4.6 ± 0.2 at 25°C

Source: This medium is available from HiMedia

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

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

Use: For the detection of fungi in routine analysis of beverages using the membrane filter technique

m-Bismuth Sulfite Broth

See: Bismuth Sulfite Broth

M-Bismuth Sulfite Broth Compositionper liter:

Peptic digest of animal tissue 20.0g Bismuth sulfite indicator 16.0g Glucose 10.0g Plant extract 10.0g

Na2HPO4 8.0g FeSO4 0.6g Brilliant Green 0.05g

pH 7.7 ± 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 and heat with frequent agitation until boiling.Boil for 1 min Do not autoclave Cool to 45°– 50°C Mix to disperse the precipitate and aseptically distribute into sterile tubes or flasks Use 2.0–2.2mL of medium for each membrane filter

Use: For the selective isolation of Salmonella typhi and other enteric bacilli and for the detection of Salmonella by the membrane filter

method

M-Bismuth Sulfite HiVeg Broth Compositionper liter:

Plant peptone 20.0g Bismuth sulfite indicator 16.0g Glucose 10.0g Plant extract 10.0g

Na2HPO4 8.0g FeSO4 0.6g Brilliant Green 0.05g

pH 7.7 ± 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 and heat with frequent agitation until boiling.Boil for 1 min Do not autoclave Cool to 45°– 50°C Mix to disperse the precipitate and aseptically distribute into sterile tubes or flasks Use 2.0–2.2mL of medium for each membrane filter

Use: For the selective isolation of Salmonella typhi and other enteric bacilli and for the detection of Salmonella by the membrane filter

method

M-Brilliant Green HiVeg Broth 1031

MBM Acetate Medium (Mineral Base Medium with Acetate)

Compositionper liter:

Agar 16.0g

NaCl 5.0g

K2HPO4 1.0g

NH4H2PO4 1.0g

Sodium acetate·3H2O 1.0g

MgSO4·7H2O 0.1g

Bromthymol Blue 0.01g

pH 6.5 ± 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 6.5 Gently

heat and bring to boiling Distribute into screw-capped tubes in 3.0mL

volumes Autoclave for 15 min at 15 psi pressure–121°C Allow tubes to

cool in a slanted position

Use: For determining the nutritional independence of bacteria

Bacte-ria that are nutritionally independent turn the medium blue

MBM Medium

See: Methylene Blue Milk Medium

MBM Medium (Modified)

(DSMZ Medium 1020) Composition per liter:

NaNO3 0.2g

KH2PO4 0.2g

NH4Cl 0.2g

MgCl2·6H2O 0.4g

KCl 0.2g

CaCl2·2H2O 0.1g

Resazurin 1.0mg

Thiosulfate solution 10.0mL

Trace elements solution SL-4 .2.0mL

pH 7.0 ± 0.2 at 25°C

Thiosulfate Solution:

Composition per10.0mL:

Na2S2O3 2.5g

Preparation of Thiosulfate Solution: Add components to

dis-tilled/deionized water and bring volume to 10.0mL Mix thoroughly

Sparge with 100% N2 Filter sterilize

Trace Elements Solution SL-4:

Compositionper liter:

EDTA 0.5g

FeSO4·7H2O 0.2g

Trace elements solution SL-6 100.0mL

Trace Elements Solution SL-6:

Compositionper 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

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

Preparation of Trace Elements Solution SL-4: Add components

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

Preparation of Medium: Add components, except thiosfulfate so-lution and trace elements SL-4 soso-lution, to distilled/deionized water and bring volume to 990.0mL Mix thoroughly Sparge solution with 80% N2 and 20% CO2 gas mixture to make it anoxic Distribute into culture vessels (e.g., 20mL in 120mL serum bottles) under a gas atmo-sphere of 80% N2 and 20% CO2 Autoclave for 15 min at 15 psi pres-sure–121°C Cool to room temperature Prior to inoculation, add the trace elements solution and thiosulfate solution to the medium Adjust

pH to 7.0 After inoculation, pressurize culture vessels to 0.5 bar over-pressure with 100% H2 gas

Use: For the cultivation of Sulfuricurvum kujiense

m-Brilliant Green Broth

See: Brilliant Green Broth

M-Brilliant Green Broth Compositionper liter:

Proteose peptone 20.0g Lactose 20.0g Saccharose 20.0g NaCl 10.0g Yeast extract 6.0g Phenol Red 0.16g Brilliant Green 0.025g

pH 6.9 ± 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 to1.0L Mix thoroughly Distribute into tubes containing inverted Durham tubes, in 10.0mL amounts for testing 1.0mL or less of sample Gently heat and bring to boiling Do not au-toclave Cool the broth rapidly Medium is sensitive to light

Use: For the detection of coliform microorganisms in foods, dairy products, water, and wastewater, as well as in other materials of sani-tary importance

M-Brilliant Green HiVeg Broth Compositionper liter:

Plant peptone No 3 20.0g Lactose 20.0g Saccharose 20.0g NaCl 10.0g Yeast extract 6.0g Phenol Red 0.16g Brilliant Green 0.025g

pH 6.9 ± 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 to1.0L Mix thoroughly Distribute into tubes containing inverted Durham tubes, in 10.0mL amounts for testing 1.0mL or less of sample Gently heat and bring to boiling Do not au-toclave Cool the broth rapidly Medium is sensitive to light

Use: For the detection of coliform microorganisms in foods, dairy

products, water, and wastewater, as well as in other materials of

sani-tary importance

M-Broth, HiVeg Compositionper liter:

Plant hydrolysate 12.5g

K2HPO4 5.0g

NaCl 5.0g

Sodium citrate 5.0g

Yeast extract 5.0g

D-Mannose 2.0g

MgSO4 0.8g

MnCl2 0.14g

FeSO4 0.04g

Polysorbate 80 0.75mL

pH 7.0 ± 0.22 at 25°C

Source: This medium, without polysorbate 80, is available as a

pre-mixed powder from HiMedia

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 detection of Salmonella in dried foods and feeds.

MCA Compositionper liter:

Carrots 200.0g

Agar 15.0g

Preparation of Medium: Peel and slice fresh carrots Place carrots

in a blender Add 500.0mL of distilled/deionized water Blend for 40

sec at high speed Filter through four layers of cheesecloth Squeeze

out juice from the residue Bring volume of filtrate to 1.0L with

dis-tilled/deionized water Add agar Mix thoroughly Gently heat and

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

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

tubes

Use: For the cultivation of Phytophthora megasperma

McBride Agar, Modified Compositionper liter:

Agar 15.0g

Glycine anhydride 10.0g

Tryptose 10.0g

NaCl 5.0g

Beef extract 3.0g

Phenylethanol 2.5g

LiCl 0.5g

Cycloheximide solution 10.0mL

pH 7.3 ± 0.2 at 25°C

Cycloheximide Solution:

Compositionper 10.0mL:

Cycloheximide 0.2g

Preparation of Cycloheximide Solution: Add cycloheximide to

distilled/deionized water and bring volume to 10.0mL Mix

thorough-ly Filter sterilize

Caution: Cycloheximide is toxic Avoid skin contact or aerosol

for-mation and inhalation

Preparation of Medium: Add components, except cycloheximide solution, 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 cycloheximide solution Mix thoroughly Pour into sterile Petri dishes

or distribute into sterile tubes

Use: For the isolation of Listeria monocytogenes from dairy products.

McBride Listeria Agar

Compositionper liter:

Agar 15.0g Glycine 10.0g Pancreatic digest of casein 5.0g Peptic digest of animal tissue 5.0g NaCl 5.0g Beef extract 3.0g Phenylethyl alcohol 2.5g LiCl 0.5g

pH 7.3 ± 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 or flasks Autoclave for 15 min at 15 psi pressure–121°C Pour into sterile Petri dishes or leave in tubes

Use: For the selective isolation of Listeria monocytogenes from

clini-cal and noncliniclini-cal specimens containing mixed flora

McBride Listeria HiVeg Agar Base

with Blood and Selective Supplement Compositionper liter:

Agar 15.0g Glycine anhydride 10.0g Plant hydrolysate No 1 10.0g NaCl 5.0g Plant extract 3.0g Phenyl ethanol 2.5g LiCl 0.5g 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 solu-tion, is available as a premixed powder from HiMedia

Caution: LiClis harmful Avoid bodily contact and inhalation of va-pors On contact with skin wash with plenty of water immediately

Selective Supplement Solution:

Compositionper 10.0mL:

Cycloheximide 0.2g

Preparation of Selective Supplement Solution: Add cyclohex-imide to distilled/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

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

McClung-Toabe Agar 1033

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

121°C Cool to 50°C Aseptically add 50.0mL defibrinated blood and

10.0mL selective supplement solution Mix thoroughly Pour into

ster-ile Petri dishes or leave in tubes

Use: For the selective isolation of Listeria monocytogenes from

clini-cal and noncliniclini-cal specimens containing mixed flora

McBride Listeria HiVeg Agar Base, Modified,

with Blood and Selective Supplement

(Modified McBride Listeria HiVeg Agar Base)

Compositionper liter:

Agar 15.0g

Glycine anhydride 10.0g

NaCl 5.0g

Plant extract 3.0g

Phenyl ethanol 2.5g

LiCl 0.5g

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

solu-tion, is available as a premixed powder from HiMedia

Caution: LiClis harmful Avoid bodily contact and inhalation of

va-pors On contact with skin wash with plenty of water immediately

Selective Supplement Solution:

Compositionper 10.0mL:

Cycloheximide 0.2g

Preparation of Selective Supplement Solution: Add

cyclohex-imide to distilled/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

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

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

121°C Cool to 50°C Aseptically add 50.0mL defibrinated blood and

10.0mL selective supplement solution Mix thoroughly Pour into

ster-ile Petri dishes or leave in tubes

Use: For the selective isolation of Listeria monocytogenes from

clini-cal and noncliniclini-cal specimens containing mixed flora

McCarthy Agar Compositionper liter:

Cornstarch 10.0g

Naladixic acid 0.015g

Colistin 0.01g

GC agar base 1.0L

pH 7.2 ± 0.2 at 25°C

GC Agar Base:

Compositionper liter:

Agar 10.0g

Pancreatic digest of casein 7.5g

Peptic digest of animal tissue 7.5g

NaCl 5.0g

K2HPO4 4.0g

Cornstarch 1.0g

KH2PO4 1.0g

Preparation of GC Agar Base: Add components to distilled/de-ionized water and bring volume to 1.0L Mix thoroughly

Preparation of Medium: To 1.0L of GC agar base, add the corn-starch Gently heat while stirring to dissolve Add the naladixic acid and colistin Mix thoroughly 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 isolation and differentiation of Gardnerella vaginalis (Haemophilus vaginalis, Corynebacterium vaginale) from

genitouri-nary specimens Bacteria that can utilize starch appear as colonies sur-rounded by a clear zone

McClung Carbon-Free Broth Compositionper liter:

NaNO3 2.0g

K2HPO4 0.8g MgSO4·7H2O 0.5g FeCl3 0.01g MnCl2·4H2O 8.0mg ZnSO4 2.0mg

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized wa-ter and bring volume to 1.0L Mix thoroughly Gently heat without boiling until salts dissolve Cool to 25°C Adjust pH to 7.2 Filter sterilize

Use: For use as a basal medium in determining the carbon assimilation capabilities of microorganisms

McClung-Toabe Agar Compositionper liter:

Proteose peptone 40.0g Agar 25.0g

Na2HPO4 5.0g Glucose 2.0g NaCl 2.0g

KH2PO4 1.0g MgSO4·7H2O 0.1g Egg yolk emulsion, 50% 100.0mL

pH 7.3 ± 0.2 at 25°C

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

Egg Yolk Emulsion, 50%:

Composition per 100.0mL:

Chicken egg yolks 11 Whole chicken egg 1 NaCl (0.9% solution) 50.0mL

Preparation of Egg Yolk Emulsion, 50%: Soak eggs with 1:100 dilution of saturated mercuric chloride solution for 1 min Crack eggs and separate yolks from whites Mix egg yolks with 1 chicken egg Beat to form emulsion Measure 50.0mL of egg yolk emulsion and add

to 50.0mL of 0.9% NaCl solution Mix thoroughly Filter sterilize Warm to 45°–50°C

Preparation of Medium: Add components, except egg yolk emul-sion, 50%, to distilled/deionized water and bring volume to 900.0mL Mix thoroughly Gently heat while stirring and bring to boiling Auto-clave for 15 min at 15 psi pressure–121°C Cool to 50°–55°C

Asepti-cally add 100.0mL of sterile egg yolk emulsion, 50% Mix thoroughly.

Pour into sterile Petri dishes in 15.0mL volumes

Use: For the isolation and cultivation of Clostridium perfringens in

foods

McClung-Toabe Agar, Modified

Compositionper liter:

Proteose peptone No 2 40.0g

Agar 20.0g

Na2HPO4 5.0g

Glucose 2.0g

NaCl 2.0g

KH2PO4 1.0g

MgSO4·7H2O 0.1g

Egg yolk emulsion, 50% 100.0mL

Hemin solution 1.0mL

pH 7.6 ± 0.2 at 25°C

Egg Yolk Emulsion, 50%:

Composition per 100.0mL:

Chicken egg yolks 11

Whole chicken egg 1

NaCl (0.9% solution) 50.0mL

Preparation of Egg Yolk Emulsion, 50%: Soak eggs with 1:100

dilution of saturated mercuric chloride solution for 1 min Crack eggs

and separate yolks from whites Mix egg yolks with 1 chicken egg

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

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

Warm to 45°–50°C

Hemin Solution:

Hemin 0.5g

NaOH (1N solution) 20.0mL

Preparation of Hemin Solution: Add hemin to 20.0mL of 1N

NaOH solution Mix thoroughly Bring volume to 100.0mL with

dis-tilled/deionized water

Preparation of Medium: Add components, except egg yolk

emul-sion, 50%, to distilled/deionized water and bring volume to 900.0mL

Mix thoroughly Gently heat while stirring and bring to boiling

Auto-clave for 15 min at 15 psi pressure–121°C Cool to 50°–55°C

Asepti-cally add 100.0mL of sterile egg yolk emulsion, 50% Mix thoroughly

Pour into sterile Petri dishes in 20.0mL volumes

Use: For the cultivation of a wide variety of anaerobic bacteria For the

differentiation of anaerobic bacteria based on lecithinase production

and lipase production Bacteria that produce lecithinase appear as

col-onies surrounded by a zone of insoluble precipitate Bacteria that

pro-duce lipase appear as colonies with a pearly iridescent sheen

McClung-Toabe Agar, Modified

Compositionper liter:

Proteose peptone No 2 40.0g

Agar 20.0g

Na2HPO4 5.0g

Glucose 2.0g

NaCl 2.0g

KH2PO4 1.0g

MgSO4·7H2O 0.1g

Neomycin 0.1g

Egg yolk emulsion, 50% 100.0mL Hemin solution 1.0mL

pH 7.6 ± 0.2 at 25°C

Egg Yolk Emulsion, 50%:

Composition per 100.0mL:

Chicken egg yolks 11 Whole chicken egg 1 NaCl (0.9% solution) 50.0mL

Preparation of Egg Yolk Emulsion, 50%: Soak eggs with 1:100 dilution of saturated mercuric chloride solution for 1 min Crack eggs and separate yolks from whites Mix egg yolks with 1 chicken egg Beat to form emulsion Measure 50.0mL of egg yolk emulsion and add

to 50.0mL of 0.9% NaCl solution Mix thoroughly Filter sterilize Warm to 45°–50°C

Hemin Solution:

Hemin 0.5g

NaOH (1N solution) 20.0mL

Preparation of Hemin Solution: Add hemin to 20.0mL of 1N

NaOH solution Mix thoroughly Bring volume to 100.0mL with dis-tilled/deionized water

Preparation of Medium: Add components, except egg yolk emul-sion, 50%, to distilled/deionized water and bring volume to 900.0mL Mix thoroughly Gently heat while stirring and bring to boiling Auto-clave for 15 min at 15 psi pressure–121°C Cool to 50°–55°C Asepti-cally add 100.0mL of sterile egg yolk emulsion, 50% Mix thoroughly Pour into sterile Petri dishes in 20.0mL volumes

Use: For the cultivation of Clostridium species For the differentiation

of Clostridium species based on lecithinase production and lipase

pro-duction Bacteria that produce lecithinase appear as colonies sur-rounded by a zone of insoluble precipitate Bacteria that produce lipase appear as colonies with a pearly iridescent sheen

McClung-Toabe Agar, Modified Compositionper liter:

Proteose peptone No 2 20.0g Agar 20.0g Yeast extract 5.0g Pancreatic digest of casein 5.0g NaCl 5.0g Sodium thioglycolate 1.0g Egg yolk emulsion, 50% 80.0mL

pH 7.6 ± 0.2 at 25°C

Egg Yolk Emulsion, 50%:

Composition per 100.0mL:

Chicken egg yolks 11 Whole chicken egg 1 NaCl (0.9% solution) 50.0mL

Preparation of Egg Yolk Emulsion, 50%: Soak eggs with 1:100 dilution of saturated mercuric chloride solution for 1 min Crack eggs and separate yolks from whites Mix egg yolks with 1 chicken egg Beat to form emulsion Measure 50.0mL of egg yolk emulsion and add

to 50.0mL of 0.9% NaCl solution Mix thoroughly Filter sterilize Warm to 45°–50°C

Preparation of Medium: Add components, except egg yolk emul-sion, 50%, to distilled/deionized water and bring volume to 920.0mL Mix thoroughly Gently heat while stirring and bring to boiling Auto-clave for 15 min at 15 psi pressure–121°C Cool to 50°–55°C