Handbook of Microbiological Media, Fourth Edition part 116 ppt

Methylophaga alcalica Agar 1145Preparation of Carbonate Solution: Add Na2CO3 to distilled/de-ionized water and bring volume to 10.0mL.. Preparation of Medium: Add components, except ace

Trang 1

Methylophaga alcalica Agar 1145

Preparation of Carbonate Solution: Add Na2CO3 to

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

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

Thiosulfate Solution:

Composition per10.0mL:

NaS2O3·5H2O 18.0g

Preparation of Thiosulfate Solution: Add NaS2O3·5H2O 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

Acetate Solution:

Composition per 10.0mL:

Sodium acetate 0.2g

Preparation of Acetate Solution: Add sodium acetate to distilled/

deionized water and bring volume to 10.0mL Mix thoroughly

Auto-clave for 15 min at 15 psi pressure–121°C Cool to room temperature

Preparation of Medium: Add components, except acetate, vitamin,

carbonate, thiosulfate, and magnesium sulfate solutions, to

distilled/de-ionized water and bring volume to 960.0mL Mix thoroughly Distribute

into closed vessels with a medium to headspace ratio of 1:5 to 1:10

Au-toclave for 15 min at 15 psi pressure–121°C Cool to room temperature

Aseptically add the acetate, vitamin, carbonate, thiosulfate, and

magne-sium sulfate solutions

Use: For the cultivation with faster growth rates of Methylonatrum

spp

Methylophaga Agar

Compositionper 103.0mL:

Agar solution 50.0mL

Mineral base, 2X 50.0mL

Solution T 2.0mL

Vitamin B12 solution 1.0mL

Methanol 0.3mL

pH 7.3 ± 0.2 at 25°C

Agar Solution:

Agar 15.0g

Preparation of Agar Solution: Add agar 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

Mineral Base, 2X:

NaCl 24.0g

MgCl2·6H2O 3.0g

MgSO4·7H2O 2.0g

CaCl2·2H2O 1.0g

KCl 0.5g

Bis-Tris buffer

(bis[2-hydroxyethyl]amino-tris[hydroxymethyl]-methane) 0.5g

Wolfe’s mineral solution 10.0mL

Preparation of Mineral Base, 2X: Add components to distilled/

deionized water and bring volume to 500.0mL Mix thoroughly Adjust

pH to 7.3 Autoclave for 15 min at 15 psi pressure–121°C Cool to

50°C

Wolfe’s Mineral Solution:

Compositionper liter:

MgSO4·7H2O 3.0g Nitrilotriacetic acid 1.5g NaCl 1.0g MnSO4·H2O 0.5g FeSO4·7H2O 0.1g CoCl2·6H2O 0.1g CaCl2 0.1g ZnSO4·7H2O 0.1g CuSO4·5H2O 0.01g AlK(SO4)2·12H2O 0.01g

H3BO3 0.01g

Na2MoO4·2H2O 0.01g

Preparation of Wolfe’s Mineral 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

Solution T:

NH4Cl 10.0g Bis-Tris buffer

KH2PO4 0.7g Ferric ammonium citrate 0.3g

Preparation of Solution T: Add components to distilled/deionized water and bring volume to 100.0mL Mix thoroughly Adjust pH to 7.3 Autoclave for 15 min at 15 psi pressure–121°C

Vitamin B 12 Solution:

Vitamin B12 1.0μg

Preparation of Vitamin B 12 Solution: Add vitamin B12 to 10.0mL

of distilled/deionized water Mix thoroughly Filter sterilize

Preparation of Medium: Aseptically mix 50.0mL of the sterile agar solution with 50.0mL of the sterile mineral base, 2X Aseptically combine sterile solution T and sterile vitamin B12 solution with the sterile mineral base Filter sterilize methanol and add to basal medium Pour into sterile Petri dishes or distribute into sterile tubes

Use: For the cultivation and maintenance of Methylophaga marina.

Methylophaga alcalica Agar

(DSMZ Medium 976) Compositionper liter:

NaCl 30.0g Agar 20.0g

KH2PO4 1.0g KNO3 1.0g MgSO4·7H2O 0.22g

Na2CO3 solution 50.0mL Methanol solution 50.0mL Trace elements solution 1.0mL

pH 9.5 ± 0.2 at 25°C

Methanol Solution:

Methanol 10.0mL

Preparation of Methanol Solution: Add methanol to distilled/de-ionized water and bring volume to 50.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C

Trang 2

1146 Methylophaga alcalica Medium

Na 2 CO 3 Solution:

Na2CO3 5.0g

Preparation of Na 2 CO 3 Solution: Add Na2CO3 to

for 15 min at 15 psi pressure–121°C

Trace Elements Solution:

Ferric citrate 30.0mg

CaCl2·2H2O 30.0mg

MgCl2·4H2O 5.0mg

ZnSO4·7H2O 5.0mg

CuSO4·5H2O 0.5mg

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 methanol

solu-tion and Na2CO3 solution, to distilled/deionized water and bring

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

Autoclave for 15 min at 15 psi pressure–121°C Cool to 55°C

Asepti-cally add 50.0mL warm sterile Na2CO3 solution and 50.0mL warm

sterile methanol solution Mix thoroughly Pour into Petri dishes or

aseptically distribute into sterile tubes

Use: For the cultivation and maintenance of Methylophaga alcalica.

Methylophaga alcalica Medium

NaCl 30.0g

KH2PO4 1.0g

KNO3 1.0g

MgSO4·7H2O 0.22g

Na2CO3 solution 50.0mL

Methanol solution 50.0mL

Trace elements solution 1.0mL

pH 9.5 ± 0.2 at 25°C

Methanol Solution:

Methanol 10.0mL

Preparation of Methanol Solution: Add methanol to

Na2CO3 5.0g

Preparation of Na 2 CO 3 Solution: Add Na2CO3 to

Ferric citrate 30.0mg

CaCl2·2H2O 30.0mg

MgCl2·4H2O 5.0mg

ZnSO4·7H2O 5.0mg

CuSO4·5H2O 0.5mg

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 methanol solu-tion and Na2CO3 solution, to distilled/deionized water and bring vol-ume to 900.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Cool to 25°C Aseptically add 50.0mL sterile Na2CO3 solution and 50.0mL sterile methanol solution Mix thoroughly Asep-tically distribute into sterile tubes or flasks

Use: For the cultivation of Methylophaga alcalica.

Methylophaga Broth

Mineral base 100.0mL Solution T 2.0mL Vitamin B12 solution 1.0mL Methanol 0.3mL

pH 7.3 ± 0.2 at 25°C

Mineral Base:

NaCl 24.0g MgCl2·6H2O 3.0g MgSO4·7H2O 2.0g CaCl2·2H2O 1.0g KCl 0.5g Bis-Tris buffer

(bis[2-hydroxyethyl]amino-tris[hydroxymethyl]-methane) 0.5g Wolfe’s mineral solution 10.0mL

Preparation of Mineral Base: Add components to distilled/deion-ized water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.3 Autoclave for 15 min at 15 psi pressure–121°C

MgSO4·7H2O 3.0g Nitrilotriacetic acid 1.5g NaCl 1.0g MnSO4·H2O 0.5g FeSO4·7H2O 0.1g CoCl2·6H2O 0.1g CaCl2 0.1g ZnSO4·7H2O 0.1g CuSO4·5H2O 0.01g AlK(SO4)2·12H2O 0.01g

H3BO3 0.01g

Na2MoO4·2H2O 0.01g

Preparation of Wolfe’s Mineral 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

Solution T:

NH4Cl 10.0g Bis-Tris buffer

KH2PO4 0.7g Ferric ammonium citrate 0.3g

Preparation of Solution T: Add components to distilled/deionized water and bring volume to 100.0mL Mix thoroughly Adjust pH to 7.3 Autoclave for 15 min at 15 psi pressure–121°C

Trang 3

Methylosarcina quisquillarum/ Methylosarcina fibrata Medium 1147

Vitamin B 12 Solution:

Vitamin B12 1.0μg

Preparation of Vitamin B 12 Solution: Add vitamin B12 to 10.0mL

of distilled/deionized water Mix thoroughly Filter sterilize

Preparation of Medium: Aseptically combine sterile solution T

and sterile vitamin B12 solution with the sterile mineral base Filter

sterilize methanol and add to basal medium Aseptically distribute into

sterile tubes or sterile flasks

Use: For the cultivation and maintenance of Methylophaga marina.

Methylophaga Medium

NaCl 25.0g

Agar 20.0g

Peptone 10.0g

Beef extract 7.0g

K2HPO4 1.0g

(NH4)2SO4 1.0g

Methanol, filter sterilized 10.0mL

pH 7.0 ± 0.2 at 25°C

Preparation of Medium: Add components, except methanol, to

distilled/deionized water and bring volume to 990.0mL Mix

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

pressure–121°C Cool to 50°–55°C Aseptically add 10.0mL of sterile

methanol Mix thoroughly Pour into sterile Petri dishes or distribute

into sterile tubes

Use: For the cultivation and maintenance of Methylophaga marina

and Methylophaga thalassica.

Methylophaga sulfidovorans Medium

NaCl 15.0g

Na2CO3 2.0g

MgSO4·7H2O 1.0g

(NH4)2SO4 0.5g

CaCl2·6H2O 0.33g

KCl 0.2g

KH2PO4 0.02g

DMS (Dimethylsulphide) 62.0mg

FeSO4·7H2O 1.0mg

Trace elements solution SL-10 1.0mL

Vitamin solution 1.0mL

pH 7.5 ± 0.3 at 25°C

Trace Elements Solution SL-10:

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 Sparge with 80% N2 + 20% CO2

Vitamin Solution:

Pyridoxine-HCl 500.0mg Nicotinic acid 200.0mg Thiamine 100.0mg

p-Aminobenzoic acid 100.0mg

Pantothenate 50.0mg Biotin 20.0mg Riboflavin 10.0mg Vitamin B12 10.0mg

Preparation of Vitamin Solution: Add components to distilled/ deionized water and bring volume to 1.0L Mix thoroughly Sparge with 80% H2 + 20% CO2

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.5 Distribute into tubes or bottles Autoclave for 15 min at 15 psi pres-sure–121°C

Use: For the cultivation of Methylophaga sulfidovorans.

Methylophaga thalassica Agar

(LMG Medium 73) Compositionper liter:

NaCl 25.0g Agar 20.0g Peptone 10.0g Lab Lemco beef extract 7.0g

K2HPO4 1.0g (NH4)2SO4 1.0g

pH 7.0 ± 0.2 at 25°C

Preparation of Medium: Add components, except methanol, to 990.0mL distilled/deionized water Mix thoroughly Gently heat and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C Cool

to 45°C Aseptically add 10.0mL sterile methanol Mix thoroughly Pour into sterile Petri dishes or distribute into sterile tubes

Use: For the cultivation of Methylophaga thalassica.

Methylosarcina quisquillarum/

Methylosarcina fibrata Medium

(DSMZ Medium 921) Compositionper 1012.1mL:

Solution 1 100.0mL Phosphate buffer 10.0mL Solution 3 1.0mL Trace elements 1.0mL Solution 2 0.1mL

pH 7.0 ± 0.2 at 25°C

Solution 1 (10X NMS Salts):

KNO3 10.0g MgSO4·6H2O 10.0g CaCl2·2H2O 2.0g

Preparation of Solution 1 (10X NMS Salts): Add components

to 700.0mL distilled/deionized water Mix thoroughly Bring volume to 1.0L with distilled/deionized water Mix thoroughly

Trang 4

1148 Methylotrophic Arthrobacter Hyphomicrobium Medium

Solution 2 (Fe EDTA):

Fe EDTA 3.8g

Preparation of Solution 2 (Fe EDTA): Add Fe EDTA to distilled/

deionized water and bring volume to 1.0L Mix thoroughly

Solution 3 (Sodium Molybdate):

Na2MoO4·4H2O 0.26g

Preparation of Solution 3 (Sodium Molybdate): Add

Na2MoO4·4H2O to distilled/deionized water and bring volume to 1.0L

Mix thoroughly

Trace Elements:

CuSO4·5H2O 100.0mg

FeSO4·7H2O 50.0mg

ZnSO4·7H2O 40.0mg

EDTA disodium salt 25.0mg

CoCl2·6H2O 5.0mg

MnCl2·4H2O 2.0mg

H3BO3 1.5mg

NiCl2·6H2O 1.0mg

Preparation of Trace Elements: Add components to

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

Phosphate Buffer:

Na2HPO4·2H2O 71.6g

KH2PO4 26.0g

Preparation of Phosphate Buffer: Add components to 800.0mL

distilled/deionized water Mix thoroughly Adjust pH to 6.8 Bring

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

121°C Cool to 55°C

Preparation of Medium: Add 100.0mL solution 1 to

distilled/de-ionized water and bring volume to 1.0L Mix thoroughly Add 1.0mL

of solution 3, 1.0mL of the trace elements, and 0.1mL of solution 2

Autoclave for 15 min at 15 psi pressure–121°C Cool to 55°C

Asepti-cally add 10.0mL phosphate buffer Mix thoroughly AseptiAsepti-cally

dis-tribute to sterile tubes or bottles

Use: For the cultivation of Methylosarcina fibrata and Methylosarcina

quisquiliarum.

Methylotrophic Arthrobacter Hyphomicrobium Medium

Na2HPO4·2H2O 7.9g

Dimethylsulfone 1.9g

KH2PO4 1.5g

NH4Cl 0.8g

MgSO4·7H2O 0.1g

pH 7.2-7.5 at 25°C

EDTA disodium salt 50.0g

NaOH 9.0g

CaCl2·2H2O 7.34g

FeSO4·7H2O 5.0g

MnCl2·4H2O 2.5g

ZnSO4·7H2O 1.0g

CoCl2·6H2O 0.5g

NH4(MoO4) 0.5g CuSO4·5H2O 0.2g

Preparation of Trace Elements Solution: Add Na2-EDTA to 400.0mL distilled/deionized water Mix thoroughly Add 9.0g NaOH Mix thoroughly Individually dissolve each of the other components in 40.0mL distilled/deionized water Add each of the other dissolved components to the EDTA solution and bring volume to 1.0L with

dis-tilled/deionized water Mix thoroughly Adjust the pH to 6.0 with 1M

NaOH

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 Hyphomicrobium sulfonivorans.

Na2HPO4·2H2O 7.9g

KH2PO4 1.5g

NH4Cl 0.8g MgSO4·7H2O 0.1g Trace elements solution 10.0mL Methanol 10.0mL

pH 7.2–7.5 at 25°C

EDTA disodium salt 50.0g NaOH 9.0g CaCl2·2H2O 7.34g FeSO4·7H2O 5.0g MnCl2·4H2O 2.5g ZnSO4·7H2O 1.0g CoCl2·6H2O 0.5g

NaOH

Na2HPO4·2H2O 7.9g

KH2PO4 1.5g Methylamine 1.0g

NH4Cl 0.8g MgSO4·7H2O 0.1g Trace elements solution 10.0mL

pH 7.2–7.5 at 25°C

Trang 5

Methylpyridine Medium 1149

NaOH 9.0g

CaCl2·2H2O 7.34g

FeSO4·7H2O 5.0g

MnCl2·4H2O 2.5g

ZnSO4·7H2O 1.0g

CoCl2·6H2O 0.5g

NH4(MoO4) 0.5g

CuSO4·5H2O 0.2g

Preparation of Trace Elements Solution: Add Na2-EDTA to

400.0mL distilled/deionized water Mix thoroughly Add 9.0g NaOH

Mix thoroughly Individually dissolve each of the other components in

40.0mL distilled/deionized water Add each of the other dissolved

com-ponents to the EDTA solution and bring volume to 1.0L with distilled/

deionized water Mix thoroughly Adjust the pH to 6.0 with 1M NaOH

Preparation of Medium: Add components to distilled/deionized

water and bring volume to 1.0L Mix thoroughly Distribute into tubes

Methylotrophic Arthrobacter Hyphomicrobium

Medium (DSMZ Medium 939) Compositionper liter:

Na2HPO4·2H2O 7.9g

KH2PO4 1.5g

Glucosel 1.0g

NH4Cl 0.8g

MgSO4·7H2O 0.1g

pH 7.2–7.5 at 25°C

NaOH 9.0g

CaCl2·2H2O 7.34g

FeSO4·7H2O 5.0g

MnCl2·4H2O 2.5g

ZnSO4·7H2O 1.0g

CoCl2·6H2O 0.5g

NH4(MoO4) 0.5g

CuSO4·5H2O 0.2g

Preparation of Trace Elements Solution: Add Na2-EDTA to

400.0mL distilled/deionized water Mix thoroughly Add 9.0g NaOH

Mix thoroughly Individually dissolve each of the other components in

40.0mL distilled/deionized water Add each of the other dissolved

components to the EDTA solution and bring volume to 1.0L with

NaOH

water and bring volume to 1.0L Mix thoroughly Distribute into tubes

Na2HPO4·2H2O 7.9g

KH2PO4 1.5g Fructose 1.0g

NH4Cl 0.8g MgSO4·7H2O 0.1g Trace elements solution 10.0mL

pH 7.2–7.5 at 25°C

EDTA disodium salt 50.0g NaOH 9.0g CaCl2·2H2O 7.34g FeSO4·7H2O 5.0g MnCl2·4H2O 2.5g ZnSO4·7H2O 1.0g CoCl2·6H2O 0.5g

NaOH

Methylpyridine Medium Compositionper 1002.0mL:

K2HPO4 0.61g

KH2PO4 0.39g KCl 0.25g Yeast extract 0.1g Wolfe’s mineral solution 10.0mL 2-Methylpyridine 1.0mL

MgSO4·7H2O 3.0g Nitrilotriacetic acid 1.5g NaCl 1.0g MnSO4·2H2O 0.5g CoCl2·6H2O 0.1g ZnSO4·7H2O 0.1g CaCl2·2H2O 0.1g FeSO4·7H2O 0.1g NiCl2·6H2O 0.025g KAl(SO4)2·12H2O 0.02g CuSO4·5H2O 0.01g

H3BO3 0.01g

Na2MoO4·2H2O 0.01g

Na2SeO3·5H2O 0.3mg

Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic acid to 500.0mL of distilled/deionized water Adjust pH to 6.5 with

Trang 6

1150 M-FC HiVeg Agar Base with Rosalic Acid

KOH Add remaining components one at a time Add

distilled/deion-ized water to 1.0L Adjust pH to 6.8

Preparation of Medium: Add components, except

2-methylpyri-dine, to distilled/deionized water and bring volume to 1.0L Mix

thor-oughly Autoclave for 15 min at 15 psi pressure–121°C Cool to room

temperature In a fume hood, aseptically add 1.0mL of

2-methylpyri-dine Mix thoroughly Aseptically distribute into sterile tubes or flasks

Use polyurethane foam closures to eliminate odors caused by

volatil-ization of 2-methylpyridine

Use: For the cultivation of Arthrobacter species.

M-FC Agar

See: FC Agar

M-FC Broth

See: FC Broth

M-FC HiVeg Agar Base with Rosalic Acid

Agar 15.0g

Lactose 12.5g

Plant hydrolysate No 1 10.0g

Plant peptone No 3 5.0g

NaCl 5.0g

Yeast extract 3.0g

Synthetic detergent No I 1.5g

Aniline Blue 0.1g

Rosolic acid solution 10.0mL

pH 7.4 ± 0.2 at 25°C

Source: This medium, without rosolic acid, is available as a premixed

powder from HiMedia

Rosolic Acid Solution:

Rosolic acid 1.0g

Preparation of Rosolic Acid Solution: Add rosolic acid to 0.2N

NaOH and bring volume to 100.0L Mix thoroughly

Preparation of Medium: Add 10.0mL rosolic acid solution to

950.0mL of distilled/deionized water Mix thoroughly Add other

com-ponents and bring volume to 1.0L with distilled/deionized water Mix

thoroughly Gently heat and bring to boiling with frequent mixing Do

not autoclave Pour into sterile Petri dishes or leave in tubes

Use: For the detection and enumeration of fecal coliforms using the

membrane filter technique

M-FC HiVeg Agar Base, Modified with Rosalic Acid

Agar 15.0g

Plant hydrolysate No 1 10.0g

Inositol 10.0g

Plant peptone No 3 5.0g

NaCl 5.0g

Yeast extract 3.0g

Synthetic detergent No I 1.5g

Aniline blue 0.1g

pH 7.4 ± 0.2 at 25°C

Source: This medium, without rosolic acid, is available as a premixed

powder from HiMedia

Rosolic acid 1.0g

Preparation of Medium: Add 10.0mL rosolic acid solution to 950.0mL of distilled/deionized water Mix thoroughly Add other com-ponents and bring volume to 1.0L with distilled/deionized water Mix thoroughly Gently heat and bring to boiling with frequent mixing Do not autoclave Pour into sterile Petri dishes or leave in tubes

Use: For the detection and enumeration of fecal coliforms using mem-brane filter technique

M-FC HiVeg Broth Base with Rosalic Acid Compositionper liter:

Lactose 12.5g Plant hydrolysate No 1 10.0g Plant peptone No 3 5.0g NaCl 5.0g Yeast extract 3.0g Synthetic detergent No I 1.5g Aniline blue 0.1g Rosolic acid solution 10.0mL

pH 7.4 ± 0.2 at 25°C

Source: This medium, without rosolic acid, is available as a premixed powder from HiMedia

Rosolic acid 1.0g

Preparation of Medium: Add 10.0mL of rosolic acid solution to 950.0mL of distilled/deionized water Mix thoroughly Add other com-ponents and bring volume to 1.0L with distilled/deionized water Mix thoroughly Gently heat and bring to boiling with frequent mixing Do not autoclave Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation of fecal coliform bacteria from waters and the enumeration of coliform bacteria using the membrane filtration method

M-FC Agar Composition per liter:

Agar 15.0g Tryptose 10.0g Inositol 10.0g Proteose peptone 5.0g NaCl 5.0g Yeast extract 3.0g Bile salts mixture 1.5g Aniline Blue 0.1g Selective supplement solution 10.0mL Rosolic acid solution 10.0mL

pH 7.4 ± 0.2 at 25°C

Source: This medium is available from HiMedia

Rosolic acid 0.1g

Trang 7

MG Medium 1151

Preparation of Rosolic Acid Solution: Add rosoloic acid to

Filter sterilize

Selective Supplement Solution:

Carbenicillin 0.05g

Preparation of Selective Supplement Solution: Add

carbenicil-lin to distilled/deionized water and bring volume to 10.0mL Mix

thor-oughly Filter sterilize

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 rapid enumeration of Klebsiella using the membrane

fil-ter technique

M-FC Agar Base Composition per liter:

Agar 15.0g

Lactose 12.5g

Tryptose 10.0g

Proteose peptone 5.0g

NaCl 5.0g

Yeast extract 3.0g

Bile salts mixture 1.5g

Aniline Blue 0.1g

pH 7.4 ± 0.2 at 25°C

Source: This medium is available from HiMedia

Rosolic acid 0.1g

Preparation of Rosolic Acid Solution: Add rosolic acidto

Filter sterilize

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

dis-solve components Do not autoclave Cool to 50°C Pour into Petri

dishes or aseptically distribute into sterile tubes

Use: For the detection and enumeration of fecal coliforms using the

membrane filter technique at 44.5°C

m-Fecal Coliform Agar

See: FC Agar

m-Fecal Coliform Agar, Modified

See: Fecal Coliform Agar, Modified

m-Fecal Coliform Broth

See: FC Broth

MG Medium Compositionper liter:

NaCl 100.0g

MgSO4·7H2O 3.45g

MgCl2·6H2O 2.75g

Sodium acetate 1.0g

KCl 0.335g

NH4Cl 0.25g CaCl2·2H2O 0.14g

K2HPO4·3H2O 0.14g Resazurin 1.0mg NaHCO3 solution 80.0mL Trimethylamine·HCl solution 20.0mL

Na2CO3 solution 10.0mL Trace elements solution 10.0mL Vitamin solution 10.0mL

L-Cysteine·HCl solution 10.0mL

Na2S·9H2O solution 10.0mL

pH 6.9 ± 0.2 at 25°C

NaHCO 3 Solution:

NaHCO3 5.0g

Preparation of NaHCO 3 Solution: Add NaHCO3 to distilled/de-ionized water and bring volume to 100.0mL Mix thoroughly Auto-clave for 15 min at 15 psi pressure–121°C

Trimethylamine·HCl Solution:

Trimethylamine·HCl 5.0g

Preparation of Trimethylamine·HCl Solution: Add trimethyl-amine·HCl to distilled/deionized water and bring volume to 20.0mL Mix thoroughly Sparge with 100% N2 Autoclave for 15 min at 15 psi pressure–121°C

Na2CO3 0.5g

Preparation of Na 2 CO 3 Solution: Add Na2CO3 to distilled/de-ionized water and bring volume to 10.0mL Mix thoroughly Sparge with 100% N2 Autoclave for 15 min at 15 psi pressure–121°C

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 CuSO4·5H2O 0.01g

H3BO3 0.01g

Na2MoO4·2H2O 0.01g

Na2SeO3·5H2O 0.3mg

Preparation of Trace Elements Solution: Add nitrilotriacetic acid to approximately 500.0mL 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

Pyridoxine·HCl 10.0mg Calcium DL-pantothenate 5.0mg Lipoic acid 5.0mg

Trang 8

1152 MG Medium

Nicotinic acid 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 Sparge with 100% N2

L -Cysteine·HCl Solution:

L-Cysteine·HCl 0.5g

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

distilled/deionized water and bring volume to 10.0mL Mix

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

Na 2 S·9H 2 O Solution:

Na2S·9H2O 0.5g

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

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

Preparation of Medium: Add components, except NaHCO3

solu-tion, trimethylamine·HCl solusolu-tion, Na2CO3 solution,vitamin solution,

L-cysteine·HCl solution, and Na2S·9H2O solution, to

distilled/deion-ized water and bring volume to 860.0mL Mix thoroughly Sparge with

100% N2 for 20 min Then sparge with 80% N2 + 20% CO2 for 10 min

Anaerobically distribute into tubes or bottles Autoclave for 15 min at

15 psi pressure–121°C Aseptically and anaerobically add 80.0mL of

sterile NaHCO3 solution, 20.0mL of sterile trimethylamine·HCl

solu-tion, 10.0mL of sterile Na2CO3 solution,10.0mL of sterile vitamin

so-lution, 10.0mL of sterile L-cysteine·HCl solution, and 10.0mL of sterile

Na2S·9H2O solution Mix thoroughly

Use: For the cultivation and maintenance of Methanohalobium

spe-cies, Methanohalophilus halophilus, and Methanohalophilus species.

MG Medium Compositionper liter:

NaCl 150.0g

MgSO4·7H2O 3.45g

MgCl2·6H2O 2.75g

Sodium acetate 1.0g

KCl 0.335g

NH4Cl 0.25g

CaCl2·2H2O 0.14g

K2HPO4·3H2O 0.14g

Resazurin 1.0mg

NaHCO3 solution 80.0mL

Trimethylamine·HCl solution 20.0mL

Na2CO3 solution 10.0mL

Vitamin solution 10.0mL

L-Cysteine·HCl solution 10.0mL

Na2S·9H2O solution 10.0mL

pH 6.9 ± 0.2 at 25°C

NaHCO3 5.0g

Preparation of NaHCO 3 Solution: Add NaHCO3 to distilled/de-ionized water and bring volume to 100.0mL Mix thoroughly Auto-clave for 15 min at 15 psi pressure–121°C

Trimethylamine·HCl Solution:

Trimethylamine·HCl 5.0g

Preparation of Trimethylamine·HCl Solution: Add trimethyl-amine·HCl to distilled/deionized water and bring volume to 20.0mL Mix thoroughly Sparge with 100% N2 Autoclave for 15 min at 15 psi pressure–121°C

Na2CO3 0.5g

Preparation of Na 2 CO 3 Solution: Add Na2CO3 to distilled/de-ionized water and bring volume to 10.0mL Mix thoroughly Sparge with 100% N2 Autoclave for 15 min at 15 psi pressure–121°C

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 CuSO4·5H2O 0.01g

H3BO3 0.01g

Na2MoO4·2H2O 0.01g

Na2SeO3·5H2O 0.3mg

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

Pyridoxine·HCl 10.0mg Calcium DL-pantothenate 5.0mg Lipoic acid 5.0mg Nicotinic acid 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 Sparge with 100% N2

L -Cysteine·HCl Solution:

L-Cysteine·HCl 0.5g

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

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

Trang 9

MH IH Agar 1153

Na 2 S·9H 2 O Solution:

Na2S·9H2O 0.5g

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

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

Preparation of Medium: Add components, except NaHCO3

solu-tion, trimethylamine·HCl solusolu-tion, Na2CO3 solution,vitamin solution, L

-cysteine·HCl solution, and Na2S·9H2O solution, to distilled/deionized

wa-ter and bring volume to 860.0mL Mix thoroughly Sparge with 100% N2

for 20 min Then sparge with 80% N2 + 20% CO2 for 10 min

Anaerobi-cally distribute into tubes or bottles Autoclave for 15 min at 15 psi

pres-sure–121°C Aseptically and anaerobically add 80.0mL of sterile NaHCO3

solution, 20.0mL of sterile trimethylamine·HCl solution, 10.0mL of sterile

Na2CO3 solution,10.0mL of sterile vitamin solution, 10.0mL of sterile L

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

thoroughly

Use: For the cultivation and maintenance of Methanohalophilus

spe-cies

MGA Agar Compositionper liter:

Agar 20.0g

Glucose 2.0g

L-Asparagine 1.0g

K2HPO4 0.5g

MgSO4·7H2O 0.5g

Trace salts solution 1.0mL

pH 7.4 ± 0.2 at 25°C

Trace Salts Solution:

FeSO4·7H2O 10.0g

CuSO4·5H2O 1.0g

MnSO4·7H2O 1.0g

ZnSO4·7H2O 1.0g

Preparation of Trace Salts Solution: Add components to

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

Ad-just pH to 8.0

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 isolation and cultivation of Actinomadura species,

Acti-nopolyspora species, Excellospora species, and Microspora species.

m-Green Yeast and Mold Broth

See: Green Yeast and Mold Broth

MGTY Agar

See: Marine Glucose Trypticase™ Yeast Extract Agar

MGTY Broth

See: Marine Glucose Trypticase™ Yeast Extract Broth

MH Agar 15%

NaCl 121.5g

Agar 20.0g

MgSO4·7H2O 14.4g MgCl2 10.5g Yeast extract 10.0g Proteose peptone no.3 5.0g KCl 3.0g Glucose 1.0g CaCl2 0.54g NaBr 39.0mg NaHCO3 solution 10.0mL

pH 7.5 ± 0.2 at 25°C

NaHCO3 0.09g

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

Preparation of Medium: Add components, except NaHCO3 solu-tion, to distilled/deionized water and bring volume to 990.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C

Aseptical-ly add 10.0mL sterile NaHCO3 solution Pour into sterile Petri dishes

or aseptically distribute into sterile tubes

Use: For cultivation and maintenance of Bacillus halophilus.

MH IH Agar Compositionper liter:

Solution A 490.0mL Solution B 490.0mL Supplement solution 20.0mL

pH 6.9 ± 0.2 at 25°C

Solution A:

Beef infusion 300.0g Acid hydrolysate of casein 17.5g Agar 17.0g Starch 1.5g

Preparation of Solution A: Add components to distilled/deionized water and bring volume to 490.0mL Mix thoroughly Gently heat and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C Cool

to 45°–50°C

Solution B:

Hemoglobin 10.0g

Preparation of Solution B: Add hemoglobin to distilled/deionized water and bring volume to 490.0mL Mix thoroughly Gently heat and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C Cool

to 45°–50°C

Supplement Solution:

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

Trang 10

1154 MH Medium

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

Preparation of Medium: Aseptically combine cooled, sterile

solu-tion A and cooled, sterile solusolu-tion B Mix thoroughly Adjust pH to 6.9

with sterile 1N HCl or sterile 1N KOH Aseptically add 20.0mL of

ster-ile supplement solution Pour into sterster-ile Petri dishes or distribute into

sterile tubes

Use: For the cultivation and differentiation of Legionella species.

MH Medium Compositionper liter:

NaCl 60.7g

Agar 20.0g

MgCl2·6H2O 15.0g

Yeast extract 10.0g

MgSO4·7H2O 7.4g

Proteose peptone No 3 5.0g

KCl 1.5g

Glucose 1.0g

CaCl2 0.27g

NaHCO3 0.45g

NaBr 0.19g

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

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

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

Use: For the cultivation and maintenance of Deleya salina and

Volca-niella eurihalina.

MH Medium Compositionper liter:

NaCl 60.7g

MgCl2·6H2O 15.0g

Yeast extract 10.0g

MgSO4·7H2O 7.4g

Proteose peptone No 3 5.0g

KCl 1.5g

Glucose 1.0g

CaCl2 0.27g

NaHCO3 0.045g

NaBr 0.019g

pH 7.2 ± 0.2 at 25°C

water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.2

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

121°C

Use: For the cultivation of Halomonas eurihalina.

MH Medium, 2%

KH2PO4 2.0g

(NH4)2SO4 2.0g

NaCl 0.5g MgSO4·7H2O 0.025g FeSO4·7H2O 2.0mg Glucose solution 20.0mL Methanol, filter sterilized 20.0mL

pH 7.0–7.5 at 25°C

Glucose Solution:

Glucose 1.5g

Preparation of Glucose Solution: Add glucose to distilled/deion-ized water and bring volume to 20.0mL Mix thoroughly Filter steril-ize

Preparation of Medium: Add components, except glucose solu-tion and methanol, to distilled/deionized water and bring volume to 960.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure– 121°C Aseptically add 20.0mL of sterile glucose solution and 20.0mL

of filter-sterilized methanol Mix thoroughly Aseptically distribute into sterile tubes or flasks

Use: For the cultivation of Methylobacillus fructoseoxidans and

Meth-ylophilus glucoseoxidans.

MH Medium, 10%

NaCl 81.0g Agar 15.0g Yeast extract 10.0g MgSO4·7H2O 9.6g MgCl2·6H2O 7.0g Proteose peptone No.3 5.0g KCl 2.0g Glucose 1.0g CaCl2 0.54g NaBr 26.0mg NaHCO3 solution 10.0mL

pH 7.5 ± 0.2 at 25°C

NaHCO3 0.06g

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

Preparation of Medium: Add components, except NaHCO3 solu-tion, to distilled/deionized water and bring volume to 990.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Cool to 45°C Aseptically add 10.0mL sterile NaHCO3 solution Pour into ster-ile Petri dishes or aseptically distribute into sterster-ile tubes

Use: For cultivation and maintenance of Chromohalobacter israelensis and Chromohalobacter canadensis.

MH Medium, 10%

NaCl 81.0g Yeast extract 10.0g MgSO4·7H2O 9.6g MgCl2·6H2O 7.0g Proteose peptone No 3 5.0g KCl 2.0g Glucose 1.0g

Định dạng
Số trang	10
Dung lượng	225,58 KB