Handbook of Microbiological Media, Fourth Edition part 170 pdf

Preparation of Medium: Add components, except bovine albumin solution and glucose solution, to distilled/deionized water and bring volume to 900.0L.. Preparation of Medium: Add component

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TB Nitrate Reduction Broth 1685

Preparation of Bovine Albumin Solution: Add bovine albumin

to distilled/deionized water and bring volume to 100.0mL Mix

thor-oughly Adjust pH to 7.8 with NaOH Store at −20°C

Preparation of Medium: Add components, except bovine albumin

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

volume to 900.0L Mix thoroughly Gently heat until boiling

Auto-clave for 15 min at 15 psi pressure–121°C Cool to 50°C Add 50.0mL

sterile glucose solution and 50.0mL bovine albumin solution Mix

thoroughly Aseptically distribute into sterile tubes

Use: For the cultivation of Mycobacterium tuberculosis.

TB HiVeg Broth Base with Bovine Serum and Glucose

Compositionper liter:

Plant peptone No 3 4.0g

Na2HPO4 2.5g

Yeast extract 2.0g

Sodium citrate 1.5g

KH2PO4 1.0g

MgSO4 0.6g

Polysorbate 80 0.5g

Glucose solution 50.0mL

Bovine serum 50.0mL

pH 7.0 ± 0.2 at 25°C

Source: This medium, without glucose or bovine albumin, is

avail-able as a premixed powder from HiMedia

Glucose Solution:

Compositionper 100.0mL:

Glucose 10.0g

Preparation of Glucose Solution: Add glucose to

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

ster-ilize

Preparation of Medium: Add components, except bovine serum

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

900.0L Mix thoroughly Gently heat until boiling Autoclave for 15

min at 15 psi pressure–121°C Cool to 50°C Add 50.0mL sterile

glu-cose solution and 50.0mL bovine serum Mix thoroughly Aseptically

distribute into sterile tubes

Use: For the cultivation of Mycobacterium tuberculosis.

TB HiVeg Broth Base without Tween™ 80

with Bovine Albumin and Glucose

Na2HPO4 2.5g

Yeast extract 2.0g

Sodium citrate 1.5g

KH2PO4 1.0g

MgSO4 0.6g

Bovine albumin solution 50.0mL

pH 7.0 ± 0.2 at 25°C

Source: This medium, without glucose or bovine albumin, is

avail-able as a premixed powder from HiMedia

Glucose 10.0g

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

Bovine Albumin Solution:

Bovine albumin fraction V 10.0g

Preparation of Bovine Albumin Solution: Add bovine albumin

to distilled/deionized water and bring volume to 100.0mL Mix thor-oughly Adjust pH to 7.8 with NaOH Store at −20°C

Preparation of Medium: Add components, except bovine albumin solution and glucose solution, to distilled/deionized water and bring volume to 900.0L Mix thoroughly Gently heat until boiling Auto-clave for 15 min at 15 psi pressure–121°C Cool to 50°C Add 50.0mL sterile glucose solution and 50.0mL bovine albumin solution Mix thoroughly Aseptically distribute into sterile tubes

Use: For the cultivation of mycobacteria when the presence of oleic

acid is undesirable For the cultivation of Mycobacterium tuberculosis

TB HiVeg Broth Base without Tween™ 80 with Bovine Serum and Glucose Compositionper liter:

Na2HPO4 2.5g Yeast extract 2.0g Sodium citrate 1.5g

KH2PO4 1.0g MgSO4 0.6g Glucose solution 50.0mL Bovine serum 50.0mL

pH 7.0 ± 0.2 at 25°C

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

Glucose 10.0g

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

Preparation of Medium: Add components, except bovine serum and glucose solution, to distilled/deionized water and bring volume to 900.0L Mix thoroughly Gently heat until boiling Autoclave for 15 min at 15 psi pressure–121°C Cool to 50°C Add 50.0mL sterile glu-cose solution and 50.0mL bovine serum Mix thoroughly Aseptically distribute into sterile tubes

Use: For the cultivation of mycobacteria when the presence of oleic

acid is undesirable For the cultivation of Mycobacterium tuberculosis

TB Nitrate Reduction Broth Compositionper 100.0mL:

Na2HPO4·12H2O 0.485g

KH2PO4 0.117g NaNO3 0.085g

pH 7.0 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized wa-ter and bring volume to 100.0mL Mix thoroughly Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C

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1686 TBAB 298 Medium

Use: For the differentiation of Mycobacterium species based on nitrate

reduction After growth of cells in appropriate medium, nitrate

tion is determined by making a suspension of cells in TB nitrate

reduc-tion broth and adding hydrochloric acid, sulfanilamide, and

N-naphyl-enendiamine Nitrate reduction turns the medium pink Mycobacterium

tuberculosis reduces nitrate and turns the medium deep pink within 1

min Mycobacterium bovis does not reduce nitrate and does not change

the medium

TBAB 298 Medium (Tryptose 298 Blood Agar Base Medium)

Tryptose Blood Agar Base:

Agar 15.0g

Tryptose 10.0g

NaCl 5.0g

Beef extract 3.0g

Thymine solution 2.0mL

D-Alanine solution 2.0mL

Streptomycin sulfate solution 1.6mL

pH 7.2 ± 0.2 at 25°C

Composition per 100.0mL:

Glucose 50.0g

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

ster-ilize

Thymine Solution:

Composition per 10.0mL:

Thymine 0.1g

Preparation of Thymine Solution: Add thymine to

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

ster-ilize

D- Alanine Solution:

Composition per 10.0mL:

D-Alanine 0.1g

Preparation of D- Alanine Solution: Add D-alanine to

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

ster-ilize

Streptomycin Sulfate Solution:

Composition per 10.0mL

Streptomycin sulfate 2.5g

Preparation of Streptomycin Sulfate Solution: Add

streptomy-cin sulfate to distilled/deionized water and bring volume to 10.0mL

Mix thoroughly Filter sterilize

Preparation of Medium: Add agar, tryptose, NaCl, and beef extract

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

Asepti-cally add 4.0mL of the sterile glucose solution, 2.0mL of the sterile

thymine solution, 2.0mL of the sterile alanine solution, and 1.6mL of the

sterile streptomycin sulfate solution Mix thoroughly Aseptically

distrib-ute into sterile tubes or flasks

Use: For the cultivation of Bacillus subtilis.

TBX Agar (Tryptone Bile X-glucuronide Agar) Compositionper liter:

Peptone 20.0g Agar 15.0g Bile salts 1.5g X-β-D-glucuronide 0.075g

pH 7.2 ± 0.2 at 25°C

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

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 Autoclave for 15 min at 15 psi pressure– 121°C Pour into sterile Petri dishes

Use: For the detection and enumeration of E coli in foodstuffs, animal food, and water without further confirmation E coli colonies are

col-ored blue-green The presence of the enzyme β-D-glucuronidase

differ-entiates most E coli sp from other coliforms E coli absorbs the

chro-mogenic substrate 5-bromo-4-chloro-3-indolyl-β-D-glucuronide The enzyme β-glucuronidase splits the bond between the chromophore 5-bromo-4-chloro-3-indolyl and the β-D-glucuronide Growth of accom-panying Gram-positive flora is largely inhibited by the use of bile salts

TBYA Agar (Tryptone Beef Yeast Extract Acetate Agar) Compositionper liter:

Agar 15.0g Pancreatic digest of casein 2.0g Beef extract 0.5g Yeast extract 0.5g Sodium actuate 0.2g

pH 7.2–7.4 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.2–7.4 Gently heat and bring to boiling Distribute into tubes or flasks Auto-clave 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 Leuconostoc species.

TC Amino Acids, HeLa 100X

See: Tissue Culture Amino Acids, HeLa 100X

TC Amino Acids, Minimal Eagle 50X

See: Tissue Culture Amino Acids, Minimal Eagle 50X

TC Dulbecco Solution

See: Tissue Culture Dulbecco Solution

TC Earle Solution

See: Tissue Culture Earle Solution

TC Hanks Solution

See: Tissue Culture Hanks Solution

TC Medium 199

See: Tissue Culture Medium 199

TC Medium Eagle, HeLa

See: Tissue Culture Medium Eagle, HeLa

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TCG Medium 1687

TC Medium Eagle with Earle BSS

See: Tissue Culture Medium Eagle

with Earle Balanced Salt Solution

TC Medium Eagle with Hanks BSS

See: Tissue Culture Medium Eagle

with Hank’s Balanced Salt Solution

TC Medium Ham F10

TC Medium NCTC 109

TC Medium RPMI #1640

See: Tissue Culture Medium RPMI #1640

TC Minimal Medium Eagle Spinner Modified MEM-S

See: Tissue Culture Minimal Medium Eagle Spinner Modified

TC Minimal Medium Eagle with Earle BSS

See: Tissue Culture Minimal Medium Eagle

with Earle Balanced Salts Solution

TC Tyrode Solution

See: Tissue Culture Tyrode Solution

TC Vitamins Minimal Eagle, 100X

See: Tissue Culture Vitamins Minimal Eagle, 100X

TCBS Agar (Thiosulfate Citrate Bile Salt Sucrose Agar)

Sucrose 20.0g

Agar 14.0g

NaCl 10.0g

Sodium citrate 10.0g

Na2S2O3 10.0g

Yeast extract 5.0g

Pancreatic digest of casein 5.0g

Peptic digest of animal tissue 5.0g

Oxgall 5.0g

Sodium cholate 3.0g

Ferric citrate 1.0g

Thymol Blue 0.04g

Bromthymol Blue 0.04g

pH 8.6 ± 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 Do not autoclave Cool to 45°–50°C Pour

into sterile Petri dishes or distribute into sterile tubes

Use: For the selective isolation of Vibrio cholerae and Vibrio

para-haemolyticus from a variety of clinical and nonclinical specimens.

TCBS HiVeg Agar Compositionper liter:

Sucrose 20.0g

Agar 15.0g

Plant peptone No 3 15.0g NaCl 10.0g Sodium citrate 10.0g

Na2S2O3 10.0g Yeast extract 6.0g Synthetic detergent No II 2.0g Ferric citrate 1.0g Thymol Blue 0.04g Bromthymol Blue 0.04g

pH 8.6 ± 0.2 at 25°C

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 into sterile Petri dishes or distribute into sterile tubes

Use: For the selective isolation of Vibrio cholerae and Vibrio

para-haemolyticus from a variety of clinical and nonclinical specimens For

the cultivation of enteropathogenic vibrios causing food poisoning

TCBS HiVeg Agar (Selective) Compositionper liter:

Sucrose 20.0g Agar 15.0g Plant special peptone 14.5g NaCl 10.0g Sodium citrate 10.0g

Na2S2O3 10.0g Yeast extract 5.0g Synthetic detergent No II 2.0g Synthetic detergent No IV 1.5g Ferric citrate 1.0g BromthymolBlue 0.04g Thymol Blue 0.04g

pH 8.6 ± 0.2 at 25°C

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 into sterile Petri dishes or distribute into sterile tubes

Use: For the selective isolation of Vibrio cholerae and other

entero-pathogenic vibrios

TCG Medium (DSMZ Medium 1009) Composition per liter:

Casitone 5.0g Glucose 4.0g Tryptone 3.0g Artificial seawater 1.0L

pH 7.5 ± 0.2 at 25°C

Artificial Seawater :

Sea salts, Instant Ocean 22.0g

Preparation of Artificial Seawater: Add Instant Ocean sea salts

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

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1688 TCH Medium

Preparation of Medium: Add components to artificial seawater

and bring volume to 1.0L Mix thoroughly Distribute to tubes or

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

Use: For the cultivation of Mechercharimyces mesophilus.

TCH Medium (Thiophene 2 Carboxylic Acid Hydrazide Medium)

Thiophene-2-carboxylic acid hydrazide 1.1mg

Middlebrook 7H10 agar base 1.0L

OADC enrichment 100.0mL

Glycerol 5.0mL

pH 6.6 ± 0.2 at 25°C

Middlebrook 7H10 Agar Base:

Agar 15.0g

Na2HPO4 1.5g

KH2PO4 1.5g

(NH4)2SO4 0.5g

L-Glutamic acid 0.5g

Sodium citrate 0.4g

Ferric ammonium citrate 0.04g

MgSO4·7H2O 0.025g

ZnSO4·7H2O 1.0mg

CuSO4·5H2O 1.0mg

Pyridoxine 1.0mg

Biotin 0.5mg

CaCl2·2H2O 0.5mg

Malachite Green 0.25mg

Preparation of Middlebrook 7H10 Agar Base: Add glycerol to

900.0mL of distilled/deionized water and add remaining components

Mix thoroughly Gently heat and bring to boiling

Middlebrook OADC Enrichment:

Bovine albumin fraction V 5.0g

Glucose 2.0g

NaCl 0.85g

Oleic acid 0.05g

Catalase 4.0mg

Source: This enrichment is available as a prepared enrichment from

BD Diagnostic Systems

Preparation of Middlebrook OADC Enrichment: Add

com-ponents to distilled/deionized water and bring volume to 100.0mL

Preparation for Medium: Combine components 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 differentiation of Mycobacterium species based on

sensi-tivity to TCH Mycobacterium bovis is inhibited by TCH

Mycobacte-rium tuberculosis and other mycobacteria are generally resistant to low

concentrations of TCH This distinguishes Mycobacterium bovis from

other nonchromogenic, slow-growing mycobacteria

TCY Agar Composition per liter:

NaCl 31.3g

Agar 15.0g

MgCl2·6H2O 10.8g

CaCl2·2H2O 1.0g Casamino acids 1.0g Tryptone 1.0g KCl 0.7g Yeast extract 0.2g

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 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 Flexibacter maritimus.

TCY Broth Composition per liter:

NaCl 31.3g Agar 15.0g MgCl2·6H2O 10.8g CaCl2·2H2O 1.0g Casamino acids 1.0g Tryptone 1.0g KCl 0.7g Yeast extract 0.2g

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 Flexibacter maritimus.

TD3 Medium (DSMZ Medium 876) Composition per 1078.0mL:

NaCl 20.0g MgCl2·6H2O 9.8g

Na2SO4 4.0g KCl 0.5g

NH4Cl 0.25g

KH2PO4 0.2g CaCl2·2H2O 0.1g Resazurin 0.5mg NaHCO3 solution 50.0mL

Na2S·9H2O solution 13.0mL Sodium caproate solution 10.0mL Selenite-tungstate solution 2.0mL Seven vitamin solution 1.0mL Vitamin solution 1.0mL Trace elements solution SL-10 1.0mL

pH 6.8 ± 0.2 at 25°C

Na 2 S·9H 2 O Solution :

Na2S·9H2O 0.6g

Preparation of Na 2 S·9H 2 O Solution: Add Na2S·9H2O to dis-tilled/deionized water and bring volume to 20.0mL Mix thoroughly Autoclave under 100% N2 for 15 min at 15 psi pressure–121°C Cool

to room temperature

NaHCO 3 Solution:

NaHCO3 5.0g

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TDN Broth 1689

Preparation of NaHCO 3 Solution: Add NaHCO3 to

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

with 80% N2 + 20% CO2 Filter sterilize

Sodium Caproate Solution:

Sodium caproate 0.5g

Preparation of Sodium Caproate Solution: Add sodium

cap-roate to distilled/deionized water and bring volume to 10.0mL Mix

thoroughly Sparge with 100% N2 Filter sterilize

Seven Vitamin Solution:

Pyridoxine hydrochloride 300.0mg

Thiamine-HCl·2H2O 200.0mg

Nicotinic acid 200.0mg

Vitamin B12 100.0mg

Calcium pantothenate 100.0mg

p-Aminobenzoic acid 80.0mg

D(+)-Biotin 20.0mg

Preparation of Seven Vitamin Solution: Add components to

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

Vitamin Solution:

Pyridoxine-HCl 10.0mg

Riboflavin 5.0mg

D-Ca-pantothenate 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 100.0mL Mix thoroughly Sparge

with 80% H2 + 20% CO2 Filter sterilize

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 Autoclave for 15 min at 15

psi pressure–121°C

Selenite-Tungstate Solution

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

Optional Supplemental Fatty Acid Mixture:

Valeric acid 0.5g Isovaleric acid 0.5g Alpha-Methylbutyric acid 0.5g Isobutyric acid 0.5g

Preparation of Optional Supplemental Fatty Acid Mixture:

Add components to 20.0mL distilled/deionized water Mix thoroughly Sparge with 100% N2 Filter sterilize

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2 gas atmosphere Add components, except NaHCO3 solu-tion, sodium caproate solusolu-tion, Na2S·9H2O solution, vitamin solution, seven vitamin solution, selenite-tungstate solution, and trace elements solution SL-10, to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.2 Sparge with 80% N2 + 20% CO2 Au-toclave for 15 min at 15 psi pressure–121°C Aseptically and anaerobi-cally add 50.0mL NaHCO3 solution, 10.0mL sodium caproate solution, 10.0mL Na2S·9H2O solution, 1.0mL vitamin solution, 1.0mL seven vi-tamin solution, 2.0mL selenite-tungstate solution, and 1.0mL trace ele-ments solution SL-10 Mix thoroughly Aseptically and anaerobically distribute into sterile tubes or bottles Growth can be stimulated by addi-tion of clarified rumen fluid or 10–20mL of a mixture of fatty acids Final

pH should be 6.7–6.9

Use: For the cultivation of unclassified bacterium DSM 13418

TDC Medium Composition per liter:

Agar 20.0g CaCO3 10.0g Glucose 5.0g

K2HPO4 1.0g MgSO4 1.0g

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 Azotobacter beijerinckii and other Azotobacter species.

TDN Broth (DSMZ Medium 1012) Composition per liter:

Pancreatic digest of gelatin 0.5g Casamino acids 0.5g Beef extract 0.3g Yeast extract 0.1g Magnesium chloride solution 10.0mL Calcium chloride solution 10.0mL

pH 7.2 ± 0.2 at 25°C

Calcium Chloride Solution:

CaCl2·2H2O 0.3g

Preparation of Calcium Chloride Solution: Add CaCl2·2H2O

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thor-1690 Tea Fungus Medium

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

temperature

Magnesium Chloride Solution:

MgCl2·6H2O 0.6g

Preparation of Magnesium Chloride Solution: Add MgCl2·6H2O

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

temperature

Preparation of Medium: Add components, except calcium

chlo-ride and magnesium chlochlo-ride solutions, to distilled/deionized water

and bring volume to 980.0mL Mix thoroughly Adjust the pH to 7.2

with NaOH Gently heat and bring to boiling Autoclave for 15 min at

15 psi pressure–121°C Cool to room temperature Aspetically add

cal-cium chloride and magnesium chloride solutions Mix thoroughly

Use: For the cultivation of Bdellovibrio bacteriovorus.

Tea Fungus Medium (DSMZ Medium 268) Composition per liter:

Sucrose 50.0g

Tea leaves, black 5.0g

Add tea and sucrose to a flask Add 1.0L freshly boiled tap water

Al-low to stand for 15 min Filter and cool to room temperature Place a

small disc of a cork stopper that has been steamed for 15 min on 2

suc-cessive days on the surface of the tea Place the inoculum onto the cork

Cover the beaker with aluminum foil and incubate

Use: For the cultivation of tea fungus

TEC Agar (m-TEC Agar) Compositionper liter:

Agar 15.0g

Lactose 10.0g

NaCl 7.5g

Proteose peptone 5.0g

K2HPO4 3.3g

Yeast extract 3.0g

KH2PO4 1.0g

Sodium lauryl sulfate 0.2g

Sodium deoxycholate 0.1g

Bromcresol Purple 0.08g

Bromphenol Red 0.08g

pH 5.0 ± 0.2 at 25°C

Di-agnostic Systems

Preparation of Medium: Add components to distilled/deionized

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

to boiling Adjust pH to 5.0 Sterilization is unnecessary Pour into

ster-ile Petri dishes or distribute into sterster-ile tubes or flasks Store at 2°–8°C

Use within 1 week

Use: For detection of Escherichia coli in recreational waters by the

mem-brane filter method This agar is used in conjunction with a urea substrate

to detect urease production After addition of the urea substrate,

Escheri-chia coli appears as yellow-yellow/brown colonies when viewed under a

fluorescent lamp

Tech Agar Compositionper liter:

Pancreatic digest of gelatin 20.0g Agar 13.6g

K2SO4·7H2O 10.0g MgCl2·6H2O 1.4g Glycerol 10.0mL

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, except glycerol, to dis-tilled/deionized water and bring volume to 990.0mL Mix thoroughly Add glycerol 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 production of pyocyanin pigment by Pseudomonas

spe-cies

Teepol Broth, Enriched (m-Teepol Broth, Enriched) Compositionper liter:

Peptone 40.0g Lactose 30.0g Yeast extract 6.0g Phenol Red 0.2g Sodium lauryl sulfate

(Teepol, 0.1% solution) 4.0mL

pH 7.4 ± 0.2 at 25°C

Use: For the enumeration of coliform organisms and Escherichia coli in

water by the membrane filter method

Teepol HiVeg Broth Compositionper liter:

Plant peptone 20.0g Lactose 10.0g NaCl 5.0g Teepol 1.0g Phenol Red 0.02g

pH 7.6 ± 0.2 at 25°C

Use: For the enumeration of coliform organisms and Escherichia coli in

water by the membrane filter method For the selective isolation and iden-tification of enteric, lactose-fermenting bacteria

Tellurite Blood Agar

See: Chocolate Tellurite Agar

Tellurite Glycine Agar Composition per liter:

Agar 17.5g Pancreatic digest of casein 10.0g Glycine 10.0g

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Tepidanaerobacter Medium 1691

Yeast extract 6.5g

D-Mannitol 5.0g

K2HPO4 5.0g

LiCl 5.0g

Enzymatic hydrolysate of soybean meal 3.5g

Chapman tellurite solution 10.0mL

pH 7.2 ± 0.2 at 25°C

Di-agnostic Systems

Caution: Lithium chloride is harmful Avoid bodily contact and

inha-lation of vapors On contact with skin wash with plenty of water

imme-diately

Chapman Tellurite Solution:

K2TeO3 1.0g

Preparation of Chapman Tellurite Solution: Add K2TeO3 to

distilled/deionized water and bring volume to 100.0mL Mix

thorough-ly Filter sterilize

Caution: Potassium tellurite is toxic

Preparation of Medium: Add components, except Chapman

tellu-rite 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 50°–55°C Aseptically add 10.0mL of

sterile Chapman tellurite solution Mix thoroughly Pour into sterile Petri

dishes or distribute into sterile tubes Allow the surface of the plates to

dry before inoculating

Use: For the isolation and cultivation of coagulase-positive

staphylo-cocci

Tellurite Glycine Agar Composition per liter:

Agar 16.0g

Pancreatic digest of casein 10.0g

Glycine 10.0g

Yeast extract 5.0g

D-Mannitol 5.0g

K2HPO4 5.0g

LiCl 5.0g

Chapman tellurite solution 20.0mL

pH 7.2 ± 0.2 at 25°C

Di-agnostic Systems.

Chapman Tellurite Solution:

K2TeO3 1.0g

Preparation of Chapman Tellurite Solution: Add K2TeO3 to

distilled/deionized water and bring volume to 100.0mL Mix

thorough-ly Filter sterilize

Caution: Potassium tellurite is toxic

Preparation of Medium: Add components, except Chapman

tellu-rite solution, to distilled/deionized water and bring volume to 980.0mL

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 20.0mL of

sterile Chapman tellurite solution Mix thoroughly Pour into sterile Petri

dishes or distribute into sterile tubes Allow the surface of the plates to

dry before inoculating

Use: For the quantitative detection of coagulase-positive staphylo-cocci from foods and other sources

Tellurite Polymyxin Egg Yolk Agar

See: TPEY Agar

TEP Uric Acid Medium Compositionper liter:

Agar 20.0g

Na2HPO4·12H2O 9.0g Uric acid 4.0g Pancreatic digest of casein 1.7g

KH2PO4 1.5g NaCl 0.5g Papaic digest of soybean meal 0.3g

K2HPO4 0.25g Glucose 0.25g MgSO4·7H2O 0.2g CaCl2 0.02g Ferric ammonium citrate 1.2mg MnCl2·4H2O 1.0mg

pH 7.2 ± 0.2 at 25°C

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

Use: For the cultivation and maintenance of Bacillus fastidiosus and

other microorganisms that can utilize uric acid as a carbon source

Tepidanaerobacter Medium

(DSMZ Medium 1051) Composition per liter:

Yeast extract 2.3g

NH4Cl 0.54g MgCl2·6H2O 0.2g CaCl2·2H2O 0.15g

KH2PO4 0.14g Resazurin 0.5mg Bicarbonate solution 10.0mL Glucose solution 10.0mL Sulfide solution 10.0mL Cysteine solution 10.0mL Trace elements solution 1.0mL Vitamin solution 2.0mL

pH 7.1 ± 0.2 at 25°C

Trace Elements Solution:

Nitrilotriacetic acid 12.8g FeCl3·6H2O 1.35g NaCl 1.0g NiCl2·6H2O 0.12g MnCl2·4H2O 0.1g CaCl2·2H2O 0.1g ZnCl2 0.1g

Na2SeO3·5H2O 0.026g CuCl2·2H2O 0.025g CoCl2·6H2O 0.024g

Na2MoO4·2H2O 0.024g

H3BO3 0.01g

Trang 8

1692 Tepidibacter Medium

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

Glucose Solution :

D-Glucose 2.2g

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

100% N2 Filter sterilize

Sulfide Solution :

Na2S·9H2O 0.3g

Preparation of Sulfide Solution: Add Na2S·9H2O to

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

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

temperature

Bicarbonate Solution :

NaHCO3 2.5g

Preparation of Bicarbonate Solution: Add NaHCO3 to distilled/

with a gas mixture of 80% N2 + 20% CO2 Filter sterilize

Cysteine Solution:

L-Cysteine-HCl·2H2O 0.3g

Preparation of Cysteine Solution: Add L-cysteine to

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

with 100% N2 Filter sterilize

Riboflavin 5.0mg

Lipoic acid 5.0mg

Biotin 2.0mg

Folic acid 2.0mg

Vitamin B12 0.1mg

Preparation of Vitamin Solution: 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 vitamin,

bicar-bonate, glucose, cysteine, and sulfide solutions, to distilled/deionized

water and bring volume to 958.0mL Mix thoroughly Gently heat and

bring to boiling Boil for 1 min Cool to room temperature while

sparg-ing with a gas mixture of 80% N2 + 20% CO2 Dispense into culture

vessels under an atmosphere of 80% N2 + 20% CO2 Autoclave for 15

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

add vitamin, bicarbonate, glucose, cysteine, and sulfide solutions Mix

thoroughly Adjust the pH to 7.0–7.2

Use: For the cultivation of Tepidanaerobacter syntrophicus.

Tepidibacter Medium

NaCl 18.0g Casein 10.0g MgCl2·6H2O 4.0g KCl 0.34g

NH4Cl 0.25g Yeast extract 0.2g

KH2PO4 0.18g CaCl2·2H2O 0.11g Fe(NH4)2(SO4)2·6H2O 0.02g Resazurin 1.0mg Bicarbonate solution 10.0mL Vitamin solution 10.0mL Sulfide solution 10.0mL Trace elements solution SL-10 1.0mL

pH 6.8 ± 0.2 at 25°C

Na2S·9H2O 0.5g

Preparation of Sulfide Solution: Add Na2S·9H2O to distilled/de-ionized water and bring volume to 10.0mL Mix thoroughly Autoclave under 100% N2 for 15 min at 15 psi pressure–121°C Cool to room temperature

NaHCO3 5.0g

Preparation of Bicarbonate Solution: Add NaHCO3 to distilled/ deionized water and bring volume to 10.0mL Mix thoroughly Sparge with gas mixture of 80% N2 + 20% CO2 Filter sterilize

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

Lipoic acid 5.0mg Biotin 2.0mg Folic acid 2.0mg Vitamin B12 0.1mg

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

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

Trang 9

Teredinobacter Medium 1693

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 100% N2 Autoclave for 15 min at 15 psi pressure–

121°C

Preparation of Medium: Add components, except vitamin,

bicar-bonate, and sulfide solutions, to distilled/deionized water and bring

volume to 970.0mL Mix thoroughly Gently heat and bring to boiling

Boil for 1 min Cool to room temperature while sparging with a gas

mixture of 80% N2 + 20% CO2 Dispense into culture vessels under an

atmosphere of 80% N2 + 20% CO2 Autoclave for 15 min at 15 psi

pressure–121°C Cool to room temperature Aspetically add vitamin,

bicarbonate, and sulfide solutions Mix thoroughly Adjust the pH to

6.6–7.0

Use: For the cultivation of Tepidibacter thalassicus and Deferribacter

autotrophicus.

Tepidibacter Medium with Peptone

NaCl 18.0g

Peptone 10.0g

MgCl2·6H2O 4.0g

KCl 0.34g

NH4Cl 0.25g

Yeast extract 0.2g

KH2PO4 0.18g

CaCl2·2H2O 0.11g

Fe(NH4)2(SO4)2·6H2O 0.02g

Resazurin 1.0mg

Bicarbonate solution 10.0mL

Vitamin solution 10.0mL

Sulfide solution 10.0mL

Trace elements solution SL-10 1.0mL

Wolfe's mineral elixir 1.0mL

pH 6.8 ± 0.2 at 25°C

Na2S·9H2O 0.5g

Preparation of Sulfide Solution: Add Na2S·9H2O to

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

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

temperature

NaHCO3 5.0g

Preparation of Bicarbonate Solution: Add NaHCO3 to distilled/

with a gas mixture of 80% N2 + 20% CO2 Filter sterilize

Riboflavin 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

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

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

Wolfe’s Mineral Elixir:

Compositionper liter: MgSO4·7H2O 30.0g NaCl 10.0g MnSO4·2H2O 5.0g (NH4)2NiSO4·6H2O 2.8g CoCl2·6H2O 1.8g ZnSO4·7H2O 1.8g FeSO4·7H2O 1.0g CaCl2·2H2O 1.0g KAl(SO4)2·12H2O 0.18g CuSO4·5H2O 0.1g

H3BO3 0.1g

Na2MoO4·2H2O 0.1g

Na2SeO4 0.1g

Na2WO4·2H2O 0.1g

Preparation of Wolfe’s Mineral Elixir: Adjust pH of 1.0L of dis-tilled/deionized water to 1.0 with dilute H2SO4 Add remaining com-ponents one at a time Mix throughly to dissolve

Preparation of Medium: Add components, except vitamin, bicar-bonate, and sulfide solutions, to distilled/deionized water and bring volume to 970.0mL Mix thoroughly Gently heat and bring to boiling Boil for 1 min Cool to room temperature while sparging with a gas mixture of 80% N2 + 20% CO2 Dispense into culture vessels under an atmosphere of 80% N2 + 20% CO2 Autoclave for 15 min at 15 psi pressure–121°C Cool to room temperature Aspetically add vitamin, bicarbonate, and sulfide solutions Mix thoroughly Adjust the pH to 6.6–7.0

Use: For the cultivation of Clostridium tepidiprofundi.

Teredinobacter Medium

Composition 1010.0mL:

Solution A 50.0mL Solution C 750.0mL Solution B 200.0mL Solution D 10.0mL

Trang 10

1694 Tergitol 7 Agar

Solution A:

K2HPO4 20.0mg

Na2CO3 20.0mg

Ferric ammonium citrate 6.0mg

EDTA 1.0mg

Trace metal mix A5 1.0mL

Trace Metal Mix A5:

H3BO3 2.86g

MnCl2·4H2O 1.81g

ZnSO4·7H2O 0.222g

Na2MoO4·2H2O 0.39g

CuSO4·5H2O 0.079g

Co(NO3)2·6H2O 49.4mg

Preparation of Trace Metal Mix A5: Add components to

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

Preparation of Solution A: Add components to distilled/deionized

water and bring volume to 50.0mL Mix thoroughly Autoclave for 15

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

Solution B:

Agar, noble 2.0g

Preparation of Solution B: Add agar to distilled/deionized water

and bring volume to 200.0mL Mix thoroughly Gently heat and bring

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

50°–55°C

Solution C:

Seawater 750.0mL

Preparation of Solution C: Autoclave for 15 min at 15 psi

pres-sure–121°C Cool to 50°–55°C

Solution D:

Cellulose, Sigmacell Type 101 1.0g

Preparation of Solution D: Add cellulose to distilled/deionized

water and bring volume to 10.0mL Mix thoroughly Autoclave for 15

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

Preparation of Medium: Aseptically combine 50.0mL of sterile

solution A, 200.0mL of sterile solution B, 750.0mL of sterile solution

C, and 10.0mL of sterile solution D Mix thoroughly Aseptically

dis-tribute into sterile tubes

Use: For the cultivation of unidentified bacterium ATCC 39867

Tergitol 7 Agar Compositionper liter:

Lactose 20.0g

Agar 13.0g

Peptone 10.0g

Yeast extract 6.0g

Meat extract 5.0g

Tergitol-7 0.1g

Bromthymol Blue 0.05g

TTC solution 5.0mL

pH 7.2 ± 0.2 at 25°C

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

Unipath

TTC Solution:

Triphenyltetrazolium chloride 0.05g

Preparation of TTC Solution: Add triphenyltetrazolium chloride

to distilled/deionized water and bring volume to 100.0mL Mix thor-oughly Filter sterilize

Preparation of Medium: Add components to distilled/deionized water and bring volume to 995.0mL Mix thoroughly Gently heat and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C Cool

to 50°C Aseptically add 5.0mL of sterile TTC solution Mix

thorough-ly Pour into sterile Petri dishes or distribute into sterile tubes

Use: For the detection and enumeration of coliforms Lactose-fer-menting bacteria appear as yellow colonies Lactose-nonferLactose-fer-menting bacteria appear as blue colonies

Tergitol 7 Agar Composition per liter:

Agar 15.0g Lactose 10.0g Yeast extract 3.0g Pancreatic digest of casein 2.5g Peptic digest of animal tissue 2.5g Tergitol 7 0.1g Bromthymol Blue 25.0mg TTC solution 3.0mL

pH 6.9 ± 0.2 at 25°C

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

TTC Solution:

Triphenyltetrazolium chloride 1.0g

Preparation of TTC Solution: Add triphenyltetrazolium chloride

to distilled/deionized water and bring volume to 100.0mL Mix thor-oughly Filter sterilize

Preparation of Medium: Add components to distilled/deionized water and bring volume to 997.0mL Mix thoroughly Gently heat and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C Cool

to 50°C Aseptically add 3.0mL of sterile TTC solution Mix

thorough-ly Pour into sterile Petri dishes or distribute into sterile tubes

Use: For the selective isolation and differentiation of coliform bacteria based on lactose fermentation Lactose-fermenting bacteria appear as yellow colonies Lactose-nonfermenting bacteria appear as blue colonies

Tergitol 7 Agar H Composition per liter:

Agar 15.0g Lactose 10.0g Yeast extract 3.0g Pancreatic digest of casein 2.5g Peptic digest of animal tissue 2.5g Ferric ammonium citrate 0.5g

Na2S2O3 0.5g Tergitol 7 0.1g Bromthymol Blue 0.025g

pH 7.2 ± 0.2

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