Handbook of Microbiological Media, Fourth Edition part 155 pdf

Preparation of Medium: Add components, except NaHCO3 solu-tion, to distilled/deionized water and bring volume to 900.0mL.. 20.0mg Preparation of Seven Vitamin Solution: Add components to

Trang 1

Salmonella Differential Agar 1535

Salinibacter ruber Medium

(DSMZ Medium 936) Compositionper liter:

NaCl 195.0g

MgSO4·7H2O 49.5g

MgCl2·6H2O 34.6g

KCl 5.0g

CaCl2·2H2O 1.25g

Yeast extract 1.0g

NaBr 0.625g

NaHCO3 0.25g

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 of Salinibacter ruber.

Salinivibrio costicola Subspecies vallismortis Medium

(DSMZ Medium 597) Composition per liter:

NaCl 25.0g

MgSO4·7H2O 9.6g

MgCl2·6H2O 7.0g

Glucose 5.0g

KCl 3.8g

Yeast extract 1.0g

CaCl2·2H2O 0.5g

K2HPO4·3H2O 0.4g

NaHCO3 solution 100.0mL

pH 7.0 ± 0.2 at 25°C

NaHCO 3 Solution:

Compositionper 100.0mL:

NaHCO3 3.0g

Preparation of NaHCO 3 Solution: Add NaHCO3 to

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

sterilize

Preparation of Medium: Add components, except NaHCO3

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

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

25°C Aseptically add 100.0mL NaHCO3 solution Mix thoroughly

Aseptically distribute into sterile tubes or flasks

Use: For the cultivation of Salinivibrio costicola subsp vallismortis.

Salmonella Chromogen Agar

(Rambach Equivalent Agar)

Compositionper liter:

Agar 15.0g

Peptone 5.0g

NaCl 5.0g

Yeast extract 2.0g

Meat extract 1.0g

Sodium deoxycholate 1.0g

pH 7.3 ± 0.2 at 25°C

Source: This medium is available from Fluka, Sigma-Aldrich

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

stirring and bring to boiling Autoclave for 15 min at 15 psi pressure– 121°C Pour into sterile Petri dishes

Use: A differential diagnostic agar for the detection of Salmonella in food, including the isolation and enumeration of Salmonella from

bivalves

Salmonella Chromogenic Agar

(OSCM) Compositionper liter:

Chromogenic mix 28.0g Agar 12.0g Special peptone 10.0g Selective supplement solution 10.0mL

pH 7.2 ± 0.2 at 25°C

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

Selective Supplement Solution:

Composition per 10.0mL:

Cefsulodin 12.0mg Novobiocin 5.0mg

Preparation of Selective Supplement Solution: Add components

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

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0mL Mix thoroughly Gently heat while stirring and bring to boiling Do not autoclave Cool quickly to 50°C Mix thoroughly Pour into sterile Petri dishes

Use: For the identification of Salmonella species and differentiation of

Salmonella spp other organisms in the family Enterobacteriaceae.

This medium combines two chromogens for the detection of

Salmo-nella spp., 5-bromo-6-chloro-3-indolyl caprylate (Magenta-caprylate)

and 5-bromo-4-chloro-3-indolyl-β-D galactopyranoside (X-gal) X-gal

is a substrate for the enzyme β-D-galactosidase Hydrolysis of the

chro-mogen, Magenta-caprylate, by lactose-negative Salmonella species

results in magenta colonies The addition of the selective supplement solution increases the selectivity of the medium Novobiocin inhibits

Proteus growth and cefsulodin inhibits growth of pseudomonads.

Salmonella Differential Agar

(RajHans Medium) Compositionper liter:

Agar 12.0g Propylene glycol 10.0g Peptone, special 8.0g B.C indicator 2.0g Yeast extract 2.0g Sodium deoxycholate 1.0g

pH 7.3 ± 0.2 at 25°C

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

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

Do not autoclave Cool to 50°C Mix thoroughly Pour into sterile Petri dishes

Trang 2

1536 Salmonella Differential HiVeg Agar, Modified

Use: A selective chromgenic medium used for the isolation and

differ-entiation of Salmonella species from the members of

Enterobacteri-aceae, especially Proteus species.

Salmonella Differential HiVeg Agar, Modified

(Salmonella Differential Agar, Modified, HiVeg)

Propylene glycol 10.0g

Plant special peptone 8.0g

NaCl 5.0g

Yeast extract 3.0g

B.C indicator 2.0g

Synthetic detergent 1.0g

pH 7.3 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from

Hi-Media

stirring and bring to boiling Mix to completely dissolve components

Do not autoclave Cool to 50°C Mix thoroughly Pour into sterile Petri

dishes

Use: A selective chromogenic medium used for the isolation and

dif-ferentiation of Salmonella species from the members of

Enterobacteri-aceae, especially Proteus species.

Salmonella HiVeg Agar, ONOZ

Agar 15.0g

Sucrose 13.0g

Lactose 11.5g

Na3-citrate·5H2O 9.3g

Plant peptone 8.625g

Plant extract No 1 6.0g

L-Phenylalanine 5.0g

Na2S2O3·5H2O 4.25g

Yeast extract 3.0g

Synthetic detergent No 1 2.0g

Na2HPO4·2H2O 1.0g

Ferric citrate 0.5g

Metachrome Yellow 0.47g

MgSO4 0.4g

Aniline Blue 0.25g

Neutral Red 0.022g

Brilliant Green 1.66mg

pH 7.4 ± 0.2 at 25°C

or flasks Gently heat and bring to boiling Do not autoclave Cool to

50°C Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation and maintenance Salmonella spp from

clini-cal specimens

Salmonella Medium

Pancreatic digest of casein 10.0g

NaCl 5.0g

pH 7.4 ± 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

Use: For the cultivation and maintenance of Escherichia coli and

Sal-monella choleraesuis.

Salmonella Rapid Test Elective Medium

Tryptone 10.0g

Na2HPO4 9.0g Sodium chloride 5.0g Casein 5.0g

KH2PO4 1.5g Malachite Green 0.0025g

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 Autoclave for 15 min

at 15 psi pressure–121°C

Use: For the Oxoid Salmonella Rapid Test which is for the

presump-tive detection of motile Salmonella in foods and environmental

sam-ples

Salmonella Rapid Test Elective Medium, 2X

Tryptone 20.0g

Na2HPO4 18.0g Sodium chloride 10.0g

KH2PO4 3.0g Casein 10.0g Malachite Green 0.005g

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 Autoclave for 15 min

at 15 psi pressure–121°C

Use: Use as described in the Oxoid Salmonella Rapid Test which is for

the presumptive detection of motile Salmonella in foods and

environ-mental samples

Salmonella Shigella Agar

(SS Agar) Composition per liter:

Agar 13.5g Lactose 10.0g Bile salts 8.5g

Na2S2O3 8.5g Sodium citrate 8.5g Beef extract 5.0g Pancreatic digest of casein 2.5g Peptic digest of animal tissue 2.5g Ferric citrate 1.0g Neutral Red 0.025g Brilliant Green 0.33mg

pH 7.0 ± 0.2 at 25°C

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

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 Do not autoclave Cool to 45°–50°C Pour

Trang 3

Salt Meat Broth 1537

into sterile Petri dishes in 20.0mL volumes Allow the surface of the

plates to dry before inoculation

Use: For the selective isolation and differentiation of pathogenic

enteric bacilli, especially those belonging to the genus Salmonella.

This medium is not recommended for the primary isolation of Shigella

species Lactose-fermenting bacteria such as Escherichia coli or

Kleb-siella pneumoniae appear as small pink or red colonies

Lactose-non-fermenting bacteria—such as Salmonella species, Proteus species, and

Shigella species—appear as colorless colonies Production of H2S by

Salmonella species turns the center of the colonies black.

Salmonella Shigella Agar, Modified

(SS Agar, Modified) Composition per liter:

Agar 12.0g

Lactose 10.0g

Sodium citrate 10.0g

Na2S2O3 8.5g

Bile salts 5.5g

Beef extract 5.0g

Peptone 5.0g

Ferric citrate 1.0g

Neutral Red 0.025g

pH 7.3 ± 0.2 at 25°C

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

Unipath

stirring and bring to boiling Do not autoclave Cool to 45°–50°C Pour

into sterile Petri dishes in 20.0mL volumes Allow the surface of the

plates to dry before inoculation

Use: For the selective isolation and differentiation of pathogenic

enteric bacilli, especially those belonging to the genus Salmonella.

This medium provides better growth of Shigella species

Lactose-fer-menting bacteria such as Escherichia coli or Klebsiella pneumoniae

appear as small pink or red colonies Lactose-nonfermenting

bacte-ria—such as Salmonella species, Proteus species, and Shigella

spe-cies—appear as colorless colonies Production of H2S by Salmonella

species turns the center of the colonies black

Salmonella Shigella Deoxycholate Agar

See: SS Deoxycholate Agar

Salt Agar Compositionper liter:

NaCl 58.4g

Agar 15.0g

Proteose peptone 5.0g

pH 6.9 ± 0.2 at 25°C

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

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

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

Use: For the cultivation and maintenance of Marinococcus halophilus.

Salt Broth, Modified Composition per liter:

NaCl 65.0g Enzymatic digest of animal tissue 10.0g Heart digest 10.0g Glucose 1.0g Bromcresol Purple 0.016g

pH 7.2 ± 0.2 at 25°C

Source: Available as a prepared medium from BD Diagnostic Sys-tems

Use: For the cultivation and differentiation of the enterococcal group

D streptococci from nonenterococcal group D streptococci based on salt tolerance

Salt Colistin Broth Compositionper liter:

NaCl 20.0g Peptone 10.0g Yeast extract 3.0g Colistin solution 10.0mL

pH 7.4 ± 0.2 at 25°C

Colistin Solution:

Colistin methane sulfonate 500,000U

Preparation of Colistin Solution: Add colistin methane sulfonate

Preparation of Medium: Add components, except colistin solu-tion, to distilled/deionized water and bring volume to 990.0mL Mix thoroughly Gently heat until dissolved Autoclave for 15 min at 15 psi pressure–121°C Cool to 25°C Aseptically add sterile colistin solu-tion Mix thoroughly Aseptically distribute into sterile tubes or flasks

Use: For the cultivation of halophilic Vibrio species.

Salt Malt Agar (SMA1) Compositionper liter:

NaCl 25.0g Agar 15.0g Malt extract 10.0g

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 cultivation of Dendryphiella salina and Dendryphiella

arenaria

Salt Meat Broth Compositionper liter:

NaCl 100.0g Neutral ox-heart tissue 30.0g Beef extract 10.0g Peptone 10.0g

pH 7.6 ± 0.2 at 25°C

Source: This medium is available as tablets from Oxoid Unipath

Trang 4

1538 Salt Medium

Use: For the isolation and cultivation of staphylococci from specimens

with a mixed flora such as fecal specimens, especially during the

inves-tigation of staphylococcal food poisoning

Salt Medium Compositionper liter:

NaCl 58.4g

Proteose peptone 5.0g

pH 4.9 ± 0.2 at 25°C

Use: For the cultivation and maintenance of Marinococcus halophilus.

Salt Nutrient Agar Compositionper liter:

NaCl 25.0g

Agar 15.0g

Peptone 5.0g

Yeast extract 2.0g

Beef extract 1.0g

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

Use: For the cultivation of Bacillus species, Pseudomonas

beijer-inckii, and other Pseudomonas species.

Salt Polymyxin Broth

(SPB) Compositionper liter:

NaCl 20.0g

Yeast extract 3.0g

Polymyxin B 250,000U

pH 8.8 ± 0.2 at 25°C

water and bring volume to 1.0L Mix thoroughly Adjust pH to 8.8

Distribute into tubes or flasks Autoclave for 10 min at 10 psi pressure–

115°C

Use: For the isolation and cultivation of Vibrio species from foods.

Salt Polymyxin HiVeg Broth Base with Polymyxin

NaCl 20.0g

Plant hydrolysate 10.0g

Yeast extract 3.0g

Selective supplement 10.0mL

pH 8.8 ± 0.2 at 25°C

Source: This medium, without selective supplement, is available as a

premixed powder from HiMedia

Selective Supplement:

Polymyxin B sulfate 100,000U

Preparation of Selective Supplement: Add polymycin B sulfate

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

or flasks Mix thoroughly Adjust pH to 8.8 Distribute into tubes or flasks Autoclave for 10 min at 10 psi pressure–115°C Cool to 50°C Aseptically add 10.0mL sterile selective supplement Mix thoroughly Pour into sterile Petri dishes or aseptically distribute into tubes

Use: For the isolation and cultivation of Vibrio species from foods.

Salt Tolerance Medium Composition per liter:

Beef heart, infusion from 500.0g NaCl 65.0g Tryptose 10.0g Glucose 1.0g Indicator solution 1.0mL

pH 7.4 ± 0.2 at 25°C

Indicator Solution:

Bromcresol Purple 1.6g Ethanol (95% solution) 100.0mL

Preparation of Indicator Solution: Add Bromcresol Purple to ethanol Mix thoroughly

Use: For the cultivation of salt-tolerant Streptococcus species and

other salt-tolerant Gram-positive cocci For the differentiation of Gram-positive cocci based on salt tolerance

Salt Tolerance Medium Compositionper liter:

NaCl 60.0g Peptone 5.0g Yeast extract 2.0g Beef extract 1.0g

pH 7.4 ± 0.2 at 25°C

Use: For the cultivation and differentiation of Aeromonas and

Ple-siomonas species based on salt tolerance.

Salt Tolerance Medium Composition per liter:

Beef heart, solids from infusion 500.0g NaCl 65.0g Tryptose 10.0g

pH 7.4 ± 0.2 at 25°C

Use: For testing the salt tolerance of a variety of microorganisms

Trang 5

Saprospira grandis Medium 1539

Salt Tolerance Medium, Gilardi

NaCl 65.0g

Agar 15.0g

Papaic digest of soybean meal 5.0g

pH 7.3 ± 0.2 at 25°C

psi pressure–121°C Do not overheat Pour into sterile Petri dishes or

leave in tubes

Use: For the cultivation and maintenance of salt-tolerant,

nonferment-ing Gram-negative bacteria For the differentiation of nonfermentnonferment-ing

Gram-negative bacteria based on salt tolerance

Salt Tolerance Medium, Tatum

NaCl 65.0g

Peptone 5.0g

Yeast extract 2.0g

Beef extract 1.0g

pH 7.4 ± 0.2 at 25°C

Use: For the cultivation of salt-tolerant, nonfermenting

Gram-nega-tive bacteria For the differentiation of nonfermenting Gram-negaGram-nega-tive

bacteria based on salt tolerance

Sanfrancisco Medium Compositionper liter:

Rye bran or wheat bran 50.0g

Fresh baker's yeast 21.0g

Fructose 7.0g

Glucose 7.0g

Maltose 7.0g

Yeast extract 7.0g

Diammonium citrate 5.0g

Sodium acetate·3H2O 5.0g

KH2PO4·3H2O 2.5g

Meat extract 2.0g

Sodium gluconate 2.0g

Tween™ 80 1.0g

L-Cysteine·HCl 0.5g

MgSO4·7H2O 0.2g

MnSO4·4H2O 0.05g

FeSO4·7H2O 0.01g

pH 5.4 ± 0.2 at 25°C

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

121°C

Use: For the cultivation of Lactobacillus sanfrancisco

SAP 1 Agar Composition per liter:

Agar 15.0g Pancreatic digest of casein 5.0g Yeast extract 5.0g Artificial seawater 1.0L

pH 7.2 ± 0.2 at 25°C

Artificial Seawater:

NaCl 24.7g MgSO4·7H2O 6.3g MgCl2·6H2O 4.6g CaCl2 1.0g KCl 0.7g NaHCO3 0.2g

Preparation of Artificial Seawater: Add components to distilled/ deionized water and bring volume to 1.0L Mix thoroughly

Preparation of Medium: Add solid components to 1.0L of artifi-cial seawater 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 isolation and cultivation of Cytophaga species,

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

SAP 2 Agar Composition per liter:

Agar 15.0g Pancreatic digest of casein 1.0g Yeast extract 1.0g Artificial seawater 1.0L

pH 7.2 ± 0.2 at 25°C

Artificial Seawater:

NaCl 24.7g MgSO4·7H2O 6.3g MgCl2·6H2O 4.6g CaCl2 1.0g KCl 0.7g NaHCO3 0.2g

Preparation of Artificial Seawater: Add components to distilled/ deionized water and bring volume to 1.0L Mix thoroughly

Preparation of Medium: Add solid components to 1.0L of artifi-cial seawater 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 isolation and cultivation of Cytophaga species,

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

Saprospira grandis Medium

Pancreatic digest of casein 5.0g Yeast extract 5.0g Ca(NO3)2·4H2O 0.1g

K2HPO4 0.02g Seawater, filtered 1.0L Trace elements 10.0mL

pH 7.0 ± 0.2 at 25°C

Trang 6

1540 Sarcina maxima Medium

Trace Elements:

FeSO4·7H2O 0.5mg

ZnSO4·7H2O 0.3mg

H3BO3 0.1mg

CoCl2·6H2O 0.1mg

CuSO4·5H2O 0.1mg

MnSO4·4H2O 0.1mg

Na2MoO4·2H2O 0.1mg

Preparation of Trace Elements: Add components to

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

Preparation of Medium: Combine components Mix thoroughly

Adjust pH to 7.0 Filter sterilize

Use: For the cultivation of Saprospira grandis.

Sarcina maxima Medium

Glucose 10.0g

Peptone 10.0g

Yeast extract 5.0g

L-Cysteine·HCl solution 10.0mL

pH 6.0 ± 0.2 at 25°C

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 Filter sterilize

Preparation of Medium: Add components, except L-cysteine·HCl

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 25°C Aseptically add sterile L

-cysteine·HCl solution Mix thoroughly Aseptically distribute into

ster-ile tubes or flasks

Use: For the cultivation of Sarcina maxima.

Sarcina Medium

(DSMZ Medium 21) Compositionper liter:

Glucose 30.0g

Peptone 5.0g

Yeast extract 5.0g

Distilled water 1000.0mL

pH 6.0 ± 0.2 at 25°C

Gently heat and bring to boiling Distribute into tubes or flasks

Auto-clave for 15 min at 15 psi pressure–121°C

Use: For the cultivation and maintenance of Sarcina ventriculi and

Sarcina maxima

Sarcina ventriculi Growth Medium

Composition per liter:

Glucose 30.0g

Peptone 5.0g

Yeast extract 5.0g

pH 6.0 ± 0.2 at 25°C

in 10.0mL volumes Autoclave for 20 min at 15 psi pressure–121°C

Use: For the cultivation and maintenance of Sarcina maxima and

Sarcina ventriculi.

Sauton’s Fluid Medium Base Composition per liter:

L-Asparagine 1.33g Polysorbate 80 0.833g Citric acid 0.66g

K2HPO4 0.177g NaH2PO4 0.056g NaCl 0.035g Ferric ammonium citrate (brown) 0.0167g Glycerol 20.0mL

pH 7.2 ± 0.2 at 25°C

Source: This medium is available from HiMedia

Preparation of Medium: Add glycerol to distilled/deionized water and bring volume to 1.0L Mix thoroughly Add remaining compo-nents Mix thoroughly Gently heat and bring to boiling Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C

Use: For the enumeration of mycobacteria

Sauton’s Medium Compositionper liter:

L-Asparagine 4.0g Citric acid 2.0g

K2HPO4 0.5g MgSO4 0.5g Triton® WR 1339 0.25g Ferric ammonium citrate 0.05g Glycerol 40.0mL

Use: For the cultivation of Mycobacterium tuberculosis strain Bacille

Calmette-Guèrin (BCG) for vaccine production

SBG Enrichment Broth (Selenite Brilliant Green Enrichment Broth) Compositionper liter:

D-Mannitol 5.0g Peptone 5.0g Yeast extract 5.0g

Na2SeO3·5H2O 4.0g

K2HPO4 2.65g

KH2PO4 1.02g Sodium taurocholate 1.0g Brilliant Green 5.0mg

pH 7.2 ± 0.2 at 25°C

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

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

Trang 7

SB/SW Medium 1541

to boiling Continue boiling for 5–10 min Do not autoclave Distribute

into sterile tubes or flasks

Use: For the selective isolation of Salmonella species, especially from

eggs and egg products

SBG Sulfa Enrichment Compositionper liter:

D-Mannitol 5.0g

Peptone 5.0g

Yeast extract 5.0g

Na2SeO3·5H2O 4.0g

K2HPO4 2.65g

KH2PO4 1.02g

Sodium taurocholate 1.0g

Sodium sulfapyridine 0.5g

pH 7.2 ± 0.2 at 25°C

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

Di-agnostic Systems

to boiling Continue boiling for 5–10 min Do not autoclave Distribute

into sterile tubes or flasks

Use: For the selective isolation of Salmonella species, especially from

eggs and egg products

SB/SW Medium Compositionper liter:

NaCl 1.0g

KCl 0.5g

MgCl2·6H2O 0.4g

NH4Cl 0.25g

KH2PO4 0.2g

CaCl2·2H2O 0.15g

Resazurin 1.0mg

Sodium crotonate solution 50.0mL

Na2S·9H2O solution 10.0mL

Seven vitamin solution 10.0mL

Sodium dithionite solution 10.0mL

Trace elements solution SL-10 1.0mL

pH 7.2 ± 0.2 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: Prepare and

dis-pense under 80% N2 + 20% CO2 Add FeCl2·4H2O to 10.0mL of HCl

solution Mix thoroughly Add distilled/deionized water and bring

vol-ume to 1.0L Add remaining components Mix thoroughly Sparge with

80% N2 + 20% CO2 Autoclave for 15 min at 15 psi pressure–121°C

Sodium Crotonate Solution:

Sodium crotonate 1.1g

Preparation of Sodium Crotonate Solution: Add sodium croto-nate to distilled/deionized water and bring volume to 50.0mL Mix thoroughly Filter sterilize Sparge with 80% N2 + 20% CO2

NaHCO3 2.5g

Preparation of NaHCO 3 Solution: Add NaHCO3 to distilled/de-ionized water and bring volume to 20.0mL Mix thoroughly Filter ster-ilize Sparge with 80% N2 + 20% CO2

Na 2 S·9H 2 O Solution:

Na2S·9H2O 0.36g

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

Seven Vitamin Solution:

Pyridoxine·HCl 0.3g Thiamine·HCl 0.2g Nicotinic acid 0.2g Calcium DL-pantothenate 0.1g Vitamin B12 0.1g

p-Aminobenzoic acid 80.0mg

Biotin 20.0mg

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

Sodium Dithionite Solution:

Sodium dithioninium 0.2g

Preparation of Sodium Dithionite Solution: Add sodium dithi-oninium to distilled/deionized water and bring volume to 10.0mL Mix thoroughly Sparge with 100% N2 Autoclave for 15 min at 15 psi pres-sure–121°C

Preparation of Medium: Prepare medium and dispense under 80%

N2 + 20% CO2 Add components, except sodium crotonate solution, seven vitamin solution, sodium dithionite solution, NaHCO3 solution, and Na2S·9H2O solution, to distilled/deionized water and bring volume

to 910.0mL Mix thoroughly Sparge with 80% N2 + 20% CO2 Auto-clave for 15 min at 15 psi pressure–121°C Aseptically and anaerobi-cally add 50.0mL of sterile sodium crotonate solution, 20.0mL of sterile NaHCO3 solution, 10.0mL of seven vitamin solution, and 10.0mL of sterile Na2S·9H2O solution Mix thoroughly Aseptically and anaerobically distribute into sterile tubes or flasks After inocula-tion, add 0.1mL of sodium dithionite solution per 10.0mL of medium

Use: For the cultivation of Syntrophobacter buswellii.

SB/SW Medium Compositionper liter:

NaCl 1.0g KCl 0.5g MgCl2·6H2O 0.4g

NH4Cl 0.25g

KH2PO4 0.2g CaCl2·2H2O 0.15g

Trang 8

1542 SC Agar

Resazurin 1.0mg

Sodium pyruvate solution 50.0mL

Na2S·9H2O solution 10.0mL

Seven vitamin solution 10.0mL

Sodium dithionite solution 10.0mL

Trace elements solution SL-10 1.0mL

pH 7.2 ± 0.2 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: Prepare and

dis-pense under 80% N2 + 20% CO2 Add FeCl2·4H2O to 10.0mL of HCl

solution Mix thoroughly Add distilled/deionized water and bring

vol-ume to 1.0L Add remaining components Mix thoroughly Sparge with

80% N2 + 20% CO2 Autoclave for 15 min at 15 psi pressure–121°C

Sodium Pyruvate Solution:

Sodium pyruvate 1.25g

Preparation of Sodium Pyruvate Solution: Add sodium

pyru-vate to distilled/deionized water and bring volume to 50.0mL Mix

thoroughly Filter sterilize Sparge with 80% N2 + 20% CO2

NaHCO3 2.5g

Preparation of NaHCO 3 Solution: Add NaHCO3 to

distilled/de-ionized water and bring volume to 20.0mL Mix thoroughly Filter

ster-ilize Sparge with 80% N2 + 20% CO2

Na 2 S·9H 2 O Solution:

Na2S·9H2O 0.36g

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

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

Seven Vitamin Solution:

Pyridoxine·HCl 0.3g

Thiamine·HCl 0.2g

Nicotinic acid 0.2g

Calcium DL-pantothenate 0.1g

Vitamin B12 0.1g

Biotin 20.0mg

Preparation of Seven Vitamin Solution: Add components to

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

Sodium Dithionite Solution:

Sodium dithioninium 0.2g

Preparation of Sodium Dithionite Solution: Add sodium dithi-oninium to distilled/deionized water and bring volume to 10.0mL Mix thoroughly Sparge with 100% N2 Autoclave for 15 min at 15 psi pres-sure–121°C

Preparation of Medium: Prepare medium and dispense under 80%

N2 + 20% CO2 Add components, except sodium pyruvate solution, seven vitamin solution, sodium dithionite solution, NaHCO3 solution, and Na2S·9H2O solution, to distilled/deionized water and bring volume

to 910.0mL Mix thoroughly Sparge with 80% N2 + 20% CO2 Auto-clave for 15 min at 15 psi pressure–121°C Aseptically and anaerobi-cally add 50.0mL of sterile sodium pyruvate solution, 20.0mL of sterile NaHCO3 solution, 10.0mL of seven vitamin solution, and 10.0mL of sterile Na2S·9H2O solution Mix thoroughly Aseptically and anaerobi-cally distribute into sterile tubes or flasks After inoculation, add 0.1mL of sodium dithionite solution per 10.0mL of medium

Use: For the cultivation of Syntrophobacter wolinii.

SC Agar (DSMZ Medium 751) Compositionper liter:

Agar 15.0g Peptone from soy meal 8.0g Corn meal, solids from infusion 2.0g

K2HPO4 1.0g

KH2PO4 1.0g MgSO4·7H2O 0.2g Hemin solution 15.0mL Serum albumin solution 10.0mL Cysteine solution 10.0mL Glucose solution 5.0mL

pH 6.6 ± 0.2 at 25°C

Hemin Solution:

Hemin 0.1g

Preparation of Hemin Solution: Add hemin to 100.0mL 0.05N

NaOH Mix thoroughly

Serum Albumin Solution:

Bovine serum albumin, fraction V 2.0g

Preparation of Serum Albumin Solution: Add bovine serum al-bumin, fraction V to distilled/deionized water and bring volume to 10.0mL Mix thoroughly Filter sterilize

Glucose Solution:

D-Glucose 1.0g

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

Cysteine Solution:

L-Cysteine·HCl·H2O 1.0g

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

thorough-ly Filter sterilize

Preparation of Medium: Add components, except glucose solu-tion, serum albumin solusolu-tion, and cysteine solusolu-tion, to distilled/deion-ized water and bring volume to 975.0mL Mix thoroughly Gently heat

Trang 9

SC Agar 1543

and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C

Cool to 50°C Aseptically add 10.0mL sterile cysteine solution,

10.0mL sterile glucose solution, and 10.0mL sterile serum albumin

so-lution Mix thoroughly Adjust pH to 6.6 Pour into sterile Petri dishes

or distribute into sterile tubes

Use: For the cultivation and maintenance of Leifsonia xyli subsp

cyno-dontis=Clavibacter xyli subsp cynodonti.

SC Agar Composition per liter:

Agar 15.0g

Papaic digest of soybean meal 8.0g

Corn meal (solids from infusion) 2.0g

K2HPO4 1.0g

KH2PO4 1.0g

MgSO4·7H2O 0.2g

Hemin solution 15.0mL

Bovine serum albumin, fraction V solution 10.0mL

L-Cysteine·H2O solution 10.0mL

Glucose solution 1.0mL

pH 6.6 ± 0.2 at 25°C

Hemin Solution:

Hemin 0.1g

NaOH (0.05N solution) 100.0mL

Preparation of Hemin Solution: Add hemin to NaOH solution

Mix thoroughly

Bovine Serum Albumin, Fraction V Solution:

Bovine serum albumin, fraction V 2.0g

Preparation of Bovine Serum Albumin, Fraction V

Solu-tion: Add bovine serum albumin to distilled/deionized water and bring

volume to 10.0mL Mix thoroughly Filter sterilize

L -Cysteine·H 2 O Solution:

L-Cysteine·H2O 1.0g

Preparation of L -Cysteine·H 2 O Solution: Add L-cysteine·H2O

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

thor-oughly Filter sterilize

D-Glucose 5.0g

Preparation of Glucose Solution: Add glucose to

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

steril-ize

Preparation of Medium: Add components—except bovine serum

albumin solution, L-cysteine·H2O solution, and glucose solution—to

thorough-ly Adjust pH to 6.6 with NaOH Gently heat and bring to boiling

Au-toclave for 15 min at 15 psi pressure–121°C Cool to 45°–50°C

Aseptically add 10.0mL of sterile bovine serum albumin solution,

10.0mL of sterile L-cysteine·H2O solution, and 1.0mL of sterile

glu-cose solution Mix thoroughly Pour into sterile Petri dishes or

distrib-ute into sterile tubes

Use: For the cultivation and maintenance of Clavibacter xyli.

SC Agar Compositionper liter:

Agar 20.0g (NH4)2SO4 5.0g

KH2PO4 1.0g MgSO4·7H2O 0.5g NaCl 0.1g CaCl2·2H2O 0.1g Inositol 2.0mg

KI 1.0mg

H3BO3 0.5mg ZnSO4·7H2O 0.4mg MnSO4·4H2O 0.4mg Thiamine·HCl 0.4mg Pyroxidine·HCl 0.4mg Niacin 0.4mg Calcium pantothenate 0.4mg

Riboflavin 0.2mg FeCl3 0.2mg

Na2MoO4·4H2O 0.2mg CuSO4·5H2O 0.04mg Folic acid 2.0μg Biotin 2.0μg Glucose solution 50.0mL Synthetic stock solution 30.0mL

Glucose 40.0g

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

Synthetic Stock Solution:

L-Isoleucine 5.25g

L-Arginine 3.48g

L-Aspartic acid 2.66g

L-Leucine 2.62g

L-Methionine 1.49g

L-Threonine 1.19g

L-Valine 1.17g

L-Serine 1.05g

L-Lysine 0.91g

L-Tryptophan 0.82g

L-Histidine 0.58g

m-Inositol 0.36g

Uracil 0.22g

L-Tyrosine 0.18g Adenine 0.135g

Preparation of Synthetic Stock Solution: Add components to distilled/deionized water and bring volume to 300.0mL Mix

thorough-ly Filter sterilize Store in the dark at 25°C

Preparation of Medium: Add components, except synthetic stock solution and glucose solution, to distilled/deionized water and bring volume to 920.0mL Mix thoroughly Gently heat and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C Cool to 50°–55°C Aseptically add 30.0mL of sterile synthetic stock solution and 50.0mL

of sterile glucose solution Mix thoroughly Pour into sterile Petri

dish-es or distribute into sterile tubdish-es

Trang 10

1544 SC Agar without Histidine

Use: For the cultivation and maintenance of Saccharomyces

cerevi-siae.

SC Agar without Histidine

Agar 20.0g

(NH4)2SO4 5.0g

KH2PO4 1.0g

MgSO4·7H2O 0.5g

NaCl 0.1g

CaCl2·2H2O 0.1g

Inositol 2.0mg

KI 1.0mg

H3BO3 0.5mg

ZnSO4·7H2O 0.4mg

MnSO4·4H2O 0.4mg

Thiamine·HCl 0.4mg

Pyroxidine·HCl 0.4mg

Niacin 0.4mg

Calcium pantothenate 0.4mg

Riboflavin 0.2mg

FeCl3 0.2mg

Na2MoO4·4H2O 0.2mg

CuSO4·5H2O 0.04mg

Folic acid 2.0μg

Biotin 2.0μg

Glucose solution 50.0mL

Synthetic stock solution 30.0mL

Glucose 40.0g

Preparation of Glucose Solution: Add glucose to

distilled/deion-ized water and bring volume to 100.0mL Mix thoroughly.Filter

ster-ilize

L-Isoleucine 5.25g

L-Arginine 3.48g

L-Leucine 2.62g

L-Methionine 1.49g

L-Threonine 1.19g

L-Valine 1.17g

L-Serine 1.05g

L-Lysine 0.91g

L-Tryptophan 0.82g

Myo-inositol 0.36g

Uracil 0.22g

L-Tyrosine 0.18g

Adenine 0.135g

Preparation of Synthetic Stock Solution: Add components to

Preparation of Medium: Add components, except synthetic stock

solution and glucose solution, to distilled/deionized water and bring

volume to 920.0mL Mix thoroughly Gently heat and bring to boiling

Autoclave for 15 min at 15 psi pressure–121°C Cool to 50°–55°C

Aseptically add 30.0mL of sterile synthetic stock solution and 50.0mL

of sterile glucose solution Mix thoroughly Pour into sterile Petri

dish-es or distribute into sterile tubdish-es

Use: For the cultivation and maintenance of strains of Saccharomyces

cerevisiae that do not require histidine.

SC Agar without Leucine and Tryptophan Compositionper liter:

Agar 20.0g (NH4)2SO4 5.0g

KH2PO4 1.0g MgSO4·7H2O 0.5g NaCl 0.1g CaCl2·2H2O 0.1g Inositol 2.0mg

KI 1.0mg

H3BO3 0.5mg ZnSO4·7H2O 0.4mg MnSO4·4H2O 0.4mg Thiamine·HCl 0.4mg Pyroxidine·HCl 0.4mg Niacin 0.4mg Calcium pantothenate 0.4mg

Riboflavin 0.2mg FeCl3 0.2mg

Na2MoO4·4H2O 0.2mg CuSO4·5H2O 0.04mg Folic acid 2.0μg Biotin 2.0μg Glucose solution 50.0mL Synthetic stock solution 30.0mL

Glucose 40.0g

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

L-Isoleucine 5.25g

L-Arginine 3.48g

L-Methionine 1.49g

L-Threonine 1.19g

L-Valine 1.17g

L-Serine 1.05g Lysine 0.91g

L-Histidine 0.58g Myo-inositol 0.36g Uracil 0.22g Adenine 0.135g

L-Tyrosine 0.18g

Preparation of Synthetic Stock Solution: Add components to distilled/deionized water and bring volume to 300.0mL Mix

Preparation of Medium: Add components, except synthetic stock solution and glucose solution, to distilled/deionized water and bring

Định dạng
Số trang	10
Dung lượng	231,54 KB