Handbook of Microbiological Media, Fourth Edition part 193 pdf

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L.. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L.

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Xanthobacter agilis Agar 1915

NH4Cl 1.0g

K2HPO4 1.0g

Plant peptone 0.78g

pH 4.8 ± 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 and bring

to boiling Boil for 1 min with mixing Distribute into tubes or flasks

Au-toclave for 15 min at 15 psi pressure–121°C Do not overheat, as this will

result in hydrolysis of the agar An additional 5.0g of agar can be used to

make a firmer agar Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation and enumeration of yeasts The low pH of the

agar selectively inhibits bacterial growth

Wort HiVeg Broth

Compositionper liter:

Malt extract 15.0g

Maltose 12.75g

Dextrin 2.75g

NH4Cl 1.0g

K2HPO4 1.0g

Plant peptone 0.78g

pH 4.8 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from

Hi-Media

to boiling Boil for 1 min with mixing Distribute into tubes or flasks

Au-toclave for 15 min at 15 psi pressure–121°C

Use: For the cultivation and enumeration of yeasts The low pH of the

agar selectively inhibits bacterial growth

Wort Sucrose Agar

Agar 25.0g

Yeast extract 1.0g

Wort solution 500.0mL

Sucrose solution 500.0mL

Wort Solution:

Compositionper 500.0mL:

Malt extract 55.0g

Preparation of Wort Solution: Add malt extract to

distilled/de-ionized water and bring volume to 500.0mL Mix thoroughly

Sucrose Solution:

Sucrose 50.0g

Preparation of Sucrose Solution: Add sucrose to

distilled/deion-ized water and bring volume to 500.0mL Mix thoroughly

Preparation of Medium: Combine components.Mix thoroughly

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 distribute into sterile tubes

Use: For the cultivation and maintenance of Aspergillus oryzae.

Wort Sucrose Broth

Yeast extract 1.0g Wort solution 500.0mL Sucrose solution 500.0mL

Wort Solution:

Malt extract 55.0g

Preparation of Wort Solution: Add malt extract to distilled/de-ionized water and bring volume to 500.0mL Mix thoroughly

Sucrose Solution:

Sucrose 50.0g

Preparation of Sucrose Solution: Add sucrose to distilled/deion-ized water and bring volume to 500.0mL Mix thoroughly

Preparation of Medium: Combine components.Mix thoroughly Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure– 121°C

Use: For the cultivation of Aspergillus oryzae.

Xanthine Agar

Solution 1 900.0mL Solution 2 100.0mL

pH 7.0 ± 0.2 at 25°C

Solution 1:

Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g

Preparation of Solution 1: Add components to distilled/deionized water and bring volume to 900.0mL Mix thoroughly Gently heat and bring

to boiling

Solution 2:

Xanthine 4.0g

Preparation of Solution 2: Add xanthine to distilled/deionized wa-ter and bring volume to 100.0mL Mix thoroughly Gently heat and bring to boiling

Preparation of Medium: Combine solutions 1 and 2 Mix thor-oughly 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 aerobic Actinomycete species Clearing around a colony indicates utilization of xanthine Streptomyces species utilize xanthine; most Nocardia and Actinomadura species do not

uti-lize xanthine

Xanthobacter agilis Agar

Solution A 1.0L Solution B 100.0mL

Solution A:

Agar 15.0g NaH2PO4·12H2O 9.0g

KH2PO4 1.5g

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1916 Xanthomonas Agar

NH4·Cl 1.0g

Sodium propionate or 3-hydroxybutyrate 1.0g

MgSO4·7H2O 0.2g

Trace elements solution 1.0mL

pH 7.0 ± 0.2 at 25°C

Trace Elements Solution:

H3BO4 560.0μg

ZnSO4·7H2O 350.0μg

NiCl2·H2O 160.0μg

Na2MoO4·2H2O 100.0μg

CuSO4·5H2O 16.0μg

MnCl2·4H2O 16.0μg

Preparation of Trace Elements Solution: Add components to

distilled/deionized water and bring volume to 2.0mL Mix thoroughly

Preparation of Solution A: Add components to distilled/deionized

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

50°–55°C

Solution B:

Ferric ammonium citrate 50.0mg

CaCl2·2H2O 100.0mg

Preparation of Solution B: Add components to distilled/deionized

water and bring volume to 100.0mL Mix thoroughly Autoclave for 15

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

Preparation of Medium: Aseptically combine 1.0L of sterile

solu-tion A with 100.0mL of sterile solusolu-tion B Mix thoroughly Pour into

sterile Petri dishes or distribute into sterile tubes

Use: For the cultivation of Xanthobacter agilis.

Xanthomonas Agar

Agar 15.0g

Pancreatic digest of gelatin 10.0g

Sucrose 10.0g

Beef extract 6.0g

pH 6.8 ± 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 Xanthomonas species.

Xanthomonas Agar

CaCO3 30.0g

Agar 15.0g

Glucose 10.0g

Yeast extract 5.0g

pH 7.0 ± 0.2 at 25°C

Cool rapidly

Use: For the cultivation and maintenance of Alcaligenes latus, Erwinia tracheiphila, Pseudomonas amygdali, Xanthomonas albilineans, Xan-thomonas axonopodis, XanXan-thomonas campestris, XanXan-thomonas fragariae, Xanthomonas maltophilia, Xanthomonas oryzae, and Xylophilus ampeli-nus.

Xanthomonas albilineans Agar

Sucrose 20.0g Agar 15.0g Peptone 10.0g Yeast extract 5.0g

pH 7.0 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized wa-ter and bring volume to 1.0L Mix thoroughly Adjust pH to 7.0 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 Xanthomonas alblinieans.

Xanthomonas maltophilia Medium

Trizma® (tris[hydroxymethyl] aminomethane) base 6.04g Glucose 5.0g KCl 1.0g NaCl 1.0g L-Phenylalanine 0.9g MgSO4 0.2g L-Arginine 0.1g L-Methionine 0.1g (NH4)2SO4 0.1g

NH4Cl 0.1g Glycerol 0.68g L-Serine 0.22g L-Alanine 0.18g

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 Adjust pH to 7.2 Fil-ter sFil-terilize Aseptically distribute into sFil-terile tubes or flasks

Use: For the cultivation of Stenotrophomonas maltophilia.

Xanthomonas Medium

Pancreatic digest of gelatin 10.0g Sucrose 10.0g Beef extract 6.0g

pH 6.8 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Gently heat with mix-ing Distribute into screw-capped test tubes Autoclave for 15 min at 15 psi pressure–121°C

Use: For the cultivation and maintenance of Xanthomonas species.

Xanthomonas TYG Agar (Xanthomonas Tryptone Yeast Extract Glucose Agar)

Composition per liter:

Agar 20.0g Pancreatic digest of casein 5.0g

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XED-AGAR 1917

Glucose 5.0g

Yeast extract 3.0g

K2HPO4 0.7g

MgSO4·7H2O 0.25g

Use: For the cultivation and maintenance of Xanthomonas species

XB45/XB90/PB90-2 Medium

(DSMZ Medium 862)

Solution A 940.0mL

Solution E 100.0mL

Solution D 10.0mL

Solution G 10.0mL

Solution F 7.2mL

Solution B 1.0mL

Solution C 1.0mL

pH 7.2 ± 0.2 at 25°C

Solution A:

NaCl 1.0g

KCl 0.5g

MgCl2·6H2O 0.4g

KH2PO4 0.2g

NH4Cl 0.25g

CaCl2·2H2O 0.15g

Resazurin 0.5mg

Preparation of Solution A: Prepare under 80% N2 + 20% CO2 gas

atmosphere Add components to distilled/deionized water and bring

vol-ume to 940.0mL Mix thoroughly Adjust pH to 7.2 Sparge with 80% N2

+ 20% CO2 Autoclave for 15 min at 15 psi pressure–121°C Cool to

25°C

Solution B:

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 Solution B: Add FeCl2·4H2O to 10.0mL of HCl

so-lution 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

Cool to 25°C

Solution C:

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 Solution C: Add components to distilled/deionized water and bring volume to 1.0L Sparge with 100% N2 Mix

thorough-ly Filter sterilize

Solution D:

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

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

Preparation of Solution D: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Sparge with 100%

N2 Filter sterilize

Solution E:

NaHCO3 5.0g

Preparation of Solution E: Add NaHCO3 to distilled/deionized water and bring volume to 100.0mL Mix thoroughly Sparge with 100% N2 Autoclave for 15 min at 15 psi pressure–121°C Cool to 25°C

Solution F:

Glucose 1.0g

Preparation of Solution F: Add glucose to distilled/deionized water and bring volume to 10.0mL Mix thoroughly Sparge with 100% N2 Auto-clave for 15 min at 15 psi pressure–121°C Cool to 25°C

Solution G :

Na2S·9H2O 0.125g

Preparation of Solution G: Add Na2S·9H2O to distilled/deionized water and bring volume to 10.0mL Mix thoroughly Autoclave under 100% N2 for 15 min at 15 psi pressure–121°C Cool to 25°C

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2 gas atmosphere Sequentially add 1.0mL solution B, 1.0mL solution C, 10.0mL solution D, 100.0mL solution E, 7.2mL so-lution F, and 10.0mL soso-lution G to 940.0mL soso-lution A Distribute an-aerobically under 80% N2 + 20% CO2 into appropriate vessels The pH should be 7.2

Use: For the cultivation of unclassified bacteria DSM 12558, DSM

12559, and DSM 12595

XED-AGAR (DSMZ Medium 1026)

Agar 18.0g Xylan 7.0g Yeast extract 3.0g

pH 7.0 ± 0.2 at 25°C

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1918 Xenorhabdus Agar

water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.0

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 Microbacterium ulmi.

Xenorhabdus Agar

Agar 15.0g

Peptone 10.0g

NaCl 5.0g

Yeast extract 5.0g

pH 7.2 ± 0.2 at 25°C

to boiling Adjust pH to 7.2 Distribute into tubes or flasks Autoclave

for 20 min at 15 psi pressure–121°C Pour into sterile Petri dishes or

leave in tubes

Use: For the cultivation and maintenance of Bacteroides galacturonicus

and Xenorhabdus nematophilus.

Xenorhabdus Broth

Peptone 10.0g

NaCl 5.0g

Yeast extract 5.0g

pH 7.2 ± 0.2 at 25°C

wa-ter and bring volume to 1.0L Mix thoroughly Adjust pH to 7.2 Distribute

into tubes or flasks Autoclave for 20 min at 15 psi pressure–121°C

Use: For the cultivation and maintenance of Bacteroides

galacturoni-cus and Xenorhabdus nematophilus.

XL Agar Base (Xylose Lysine Agar Base)

Agar 13.5g

Lactose 7.5g

Sucrose 7.5g

L-Lysine 5.0g

NaCl 5.0g

Xylose 3.5g

Yeast extract 3.0g

Phenol Red 0.08g

Thiosulfate-citrate solution 20.0mL

pH 7.5 ± 0.2 at 25°C

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

Di-agnostic Systems

Thiosulfate-Citrate Solution:

Na2S2O3 34.0g

Ferric ammonium citrate 4.0g

Preparation of Thiosulfate-Citrate Solution: Add components

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

thor-oughly

Preparation of Medium: Add components, except

thiosulfate-cit-rate solution, to distilled/deionized water and bring volume to

980.0mL Mix thoroughly Gently heat while stirring and bring to boil-ing Distribute into tubes or flasks Autoclave for 10 min at 14 psi pres-sure–118°C Cool to 55°C Aseptically add 20.0 mL of the sterile thio-sulfate-citrate solution Mix thoroughly Pour into sterile Petri dishes or leave in tubes

Use: For the isolation, cultivation, and differentiation of enteric patho-gens Nonfermenting xylose/lactose/sucrose bacteria appear as red col-onies Xylose-fermenting, lysine-decarboxylating bacteria appear as red colonies Xylose-fermenting, lysine-nondecarboxylating bacteria appear as opaque yellow colonies Lactose- or sucrose-fermenting bac-teria appear as yellow colonies

XL Agar Base

Agar 15 g Lactose 7.5g Sucrose 7.5g L-Lysine 5.0g NaCl 5.0g Xylose 3.75g Yeast extract 3.0g Phenol Red 0.08g Thiosulfate-citrate solution 20.0mL

pH 7.4 ± 0.2 at 25°C

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

Thiosulfate-Citrate Solution:

Na2S2O3 34.0g Ferric ammonium citrate 4.0g

Preparation of Thiosulfate-Citrate Solution: Add components

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

Preparation of Medium: Add components, except thiosulfate-cit-rate solution, to distilled/deionized water and bring volume to 980.0mL Mix thoroughly Gently heat while stirring and bring to boil-ing Distribute into tubes or flasks Autoclave for 10 min at 14 psi pres-sure–118°C Cool to 55°C Aseptically add 20.0 mL of the sterile thio-sulfate-citrate solution Mix thoroughly Pour into sterile Petri dishes or leave in tubes

Use: For the isolation, cultivation, and differentiation of enteric patho-gens Nonfermenting xylose/lactose/sucrose bacteria appear as red col-onies Xylose-fermenting, lysine-decarboxylating bacteria appear as red colonies Xylose-fermenting, lysine-nondecarboxylating bacteria appear as opaque yellow colonies Lactose- or sucrose-fermenting bac-teria appear as yellow colonies

XLD Agar (Xylose Lysine Deoxycholate Agar)

Agar 13.5g Lactose 7.5g Sucrose 7.5g

Na2S2O3 6.8g L-Lysine 5.0g NaCl 5.0g Xylose 3.5g

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XLT4 HiVeg Agar Base 1919

Yeast extract 3.0g

Sodium desoxycholate 2.5g

Phenol Red 0.08g

pH 7.5 ± 0.2 at 25°C

Di-agnostic Systems and Oxoid Unipath

to boiling Do not overheat Distribute into tubes or flasks Autoclave

leave in tubes Plates should be poured as soon as possible to avoid

pre-cipitation

Use: For the isolation and differentiation of enteric pathogens,

espe-cially Shigella and Providencia species Nonfermenting

xylose/lac-tose/sucrose bacteria appear as red colonies Xylose-fermenting,

lysine-decarboxylating bacteria appear as red colonies Xylose-fermenting,

lysine-nondecarboxylating bacteria appear as opaque yellow colonies

Lactose- or sucrose-fermenting bacteria appear as yellow colonies

XLD Agar (Xylose Lysine Deoxycholate Agar)

(BAM M179)

Agar 15.0g

Lactose 7.5g

Sucrose 7.5g

Na2S2O3 6.8g

L-Lysine 5.0g

NaCl 5.0g

Xylose 3.75g

Yeast extract 3.0g

Sodium deoxycholate 2.5g

Phenol Red 0.08g

pH 7.5 ± 0.2 at 25°C

Di-agnostic Systems and Oxoid

to boiling Do not overheat Distribute into tubes or flasks Autoclave

leave in tubes Plates should be poured as soon as possible to avoid

pre-cipitation

xylose/lac-tose/sucrose bacteria appear as red colonies Xylose-fermenting,

lysine-decarboxylating bacteria appear as red colonies

Xylose-fer-menting, lysine-nondecarboxylating bacteria appear as opaque yellow

colonies Lactose- or sucrose-fermenting bacteria appear as yellow

col-onies

XLD Agar, HiVeg (Xylose Lysine Deoxycholate HiVeg Agar)

Agar 15.0g

Lactose 7.5g

Sucrose 7.5g

Na2S2O3 6.8g L-Lysine 5.0g NaCl 5.0g Yeast extract 4.0g Xylose 3.5g Synthetic detergent No III 1.5g Ferric ammonium citrate 0.8g Phenol Red 0.08g Selective supplement solution 4.6mL

pH 7.4 ± 0.2 at 25°C

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

Selective Supplement Solution:

Tergitol™ 4 Proprietary

Preparation of Selective Supplement Solution: Available as pre-mixed solution

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

to boiling Do not overheat Distribute into tubes or flasks Do not au-toclave Pour into sterile Petri dishes or leave in tubes Plates should be poured as soon as possible to avoid precipitation

xylose/lac-tose/sucrose bacteria appear as red colonies Xylose-fermenting, lysine-decarboxylating bacteria appear as red colonies Xylose-fer-menting, lysine-nondecarboxylating bacteria appear as opaque yellow colonies Lactose- or sucrose-fermenting bacteria appear as yellow col-onies

XLT4 HiVeg Agar Base

Agar 18.0g Lactose 7.5g Saccharose 7.5g

Na2S2O3 6.8g L-Lysine 5.0g NaCl 5.0g Xylose 3.75g Yeast extract 3.0g Plant peptone No 3 1.6g Ferric ammonium citrate 0.8g Phenol Red 0.08g Selective supplement solution 4.6mL

pH 7.4 ± 0.2 at 25°C

Source: This medium, without selective supplement solution, is avail-able as a premixed powder from HiMedia

Preparation of Selective Supplement Solution: Available as pre-mixed solution

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

to boiling Do not overheat Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C Pour into sterile Petri dishes or leave in tubes Plates should be poured as soon as possible to avoid pre-cipitation

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1920 XPS Agar

espe-cially Shigella and Providencia species

XPS Agar

Solution A 500.0mL

Solution B 500.0mL

pH 5.1 ± 0.2 at 25°C

Solution A:

Composition per 500.0mL:

Potatoes, infusion from 40.0g

Sucrose 15.0g

Peptone 5.0g

Glucose 4.0g

Casamino acids 1.0g

Na2HPO4 0.79g

Ca(NO3)2·4H2O solution 10.0mL

Potatoes, Infusion From:

Composition per 500.0mL:

Potatoes 4.0g

Preparation of Potatoes, Infusion From: Peel and dice potatoes

Add 400.0mL of distilled/deionized water Gently heat and bring to

boiling Continue boiling for 30 min Filter through cheesecloth

Ca(NO3) 2 ·4H 2 O Solution:

Ca(NO3)2·4H2O 0.5g

Preparation of Ca(NO 3 ) 2 ·4H 2 O Solution: Add Ca(NO3)2·4H2O

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

Filter sterilize

Preparation of Solution A : Add components, except Ca(NO3)2·4H2O

solution, to distilled/deionized water and bring volume to 490.0mL

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

Asep-tically add 10.0mL of sterile Ca(NO3)2·4H2O solution Mix

thorough-ly Cool to 50°–55°C

Solution B:

Composition per 500.0mL:

Agar 20.0g

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

and bring volume to 500.0mL Mix thoroughly Gently heat and bring

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

50°–55°C

Preparation of Medium: Aseptically combine 500.0mL of

solu-tion A with 500.0mL of solusolu-tion B Mix thoroughly Aseptically pour

into sterile Petri dishes or distribute into sterile tubes

Use: For the cultivation of Xanthomonas campestris

XPS Broth

Potatoes, infusion from 40.0g

Sucrose 15.0g

Peptone 5.0g

Glucose 4.0g

Casamino acids 1.0g

Na2HPO4 0.79g

Ca(NO3)2·4H2O solution 10.0mL

pH 5.1 ± 0.2 at 25°C

Potatoes 4.0g

Preparation of Potatoes, Infusion From: Peel and dice potatoes Add 500.0mL of distilled/deionized water Gently heat and bring to boiling Continue boiling for 30 min Filter through cheesecloth

Ca(NO3)2·4H2O 0.5g

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

Preparation of Medium: Add components, except Ca(NO3)2·4H2O solution, to distilled/deionized water and bring volume to 990.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Asep-tically add 10.0mL of sterile Ca(NO3)2·4H2O solution Mix

thorough-ly Aseptically distribute into sterile tubes or flasks

Use: For the cultivation of Xanthomonas campestris.

XPS Broth with Thymidine (Thymidine Auxotroph XPS Medium)

Potatoes, infusion from 40.0g Sucrose 15.0g Peptone 5.0g Glucose 4.0g Casamino acids 1.0g

Na2HPO4 0.79g Thymidine 10.0mg Ca(NO3)2·4H2O solution 10.0mL

pH 5.1 ± 0.2 at 25°C

Potatoes 4.0g

Preparation of Potatoes, Infusion From: Peel and dice potatoes Add 500.0mL of distilled/deionized water Gently heat and bring to boiling Continue boiling for 30 min Filter through cheesecloth

Ca(NO3)2·4H2O 0.5g

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

Preparation of Medium: Add components, except Ca(NO3)2·4H2O solution, to distilled/deionized water and bring volume to 990.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–121°C Asep-tically add 10.0mL of sterile Ca(NO3)2·4H2O solution Mix

thorough-ly Aseptically distribute into sterile tubes or flasks

Use: For the cultivation of Xanthomonas oryzae.

XSM Agar

Agar 15.0g Glucose 5.0g Sucrose 2.0g Malt extract 1.0g

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Xylella Agar 1921

Yeast extract 1.0g

Liver extract concentrate 1.0g

Corn steep liquor 1.0g

pH 7.0 ± 0.2 at 25°C

Preparation of Medium: Add components to tap water and bring

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

Dis-tribute into screw-capped test tubes 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 Streptomyces cinereus

and Streptomyces flaveus.

Xylan Medium

Xylan 30.0g

Agar 12.0g

Peptone 2.0g

Yeast extract 0.5g

L-Cysteine·HCl·H2O 0.25g

Na2S·9H2O 0.25g

Rumen fluid 400.0mL

NaHCO3 solution 40.0mL

Mineral solution I 25.0mL

Mineral solution II 25.0mL

Wolfe’s vitamin solution 10.0mL

VFA solution 10.0mL

Hemin solution 10.0mL

Trace elements solution SL-6 1.0mL

pH 7.0 ± 0.2 at 25°C

NaHCO 3 Solution:

NaHCO3 3.96g

Preparation of NaHCO 3 Solution: Add NaHCO3 to

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

100% CO2

Mineral Solution I:

K2HPO4 3.0g

Preparation of Mineral Solution I: Add K2HPO4 to

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

Mineral Solution II:

Sodium citrate 20.0g

NaCl 12.0g

KH2PO4 6.0g

MgCl2·6H2O 2.0g

CaCl2 1.2g

Preparation of Mineral Solution II: Add components to

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

Wolfe’s Vitamin Solution:

Pyridoxine·HCl 10.0mg

Thiamine·HCl 5.0mg

Riboflavin 5.0mg

Nicotinic acid 5.0mg

Calcium pantothenate 5.0mg

Thioctic acid 5.0mg

Biotin 2.0mg Folic acid 2.0mg Cyanocobalamin 100.0μg

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

VFA Solution:

Acetic acid 178.3mL Propionic acid 59.6mL

n-Butyric acid 38.4mL

Isobutyric acid 9.5mL

n-Valeric acid 9.4mL

Isovaleric acid 9.3mL DL-α-Methylbutyric acid 4.4mL

Preparation of VFA Solution: Add components to distilled/deion-ized water and bring volume to approximately 500.0mL Adjust pH to 7.5 with NaOH Mix thoroughly Bring volume to 1.0L with distilled/ deionized water

Hemin Solution:

Hemin 0.01g

Preparation of Hemin Solution: Add hemin to 100.0mL of 0.01N

NaOH Mix thoroughly

Trace Elements Solution SL-6:

H3BO3 0.3g CoCl2·6H2O 0.2g ZnSO4·7H2O 0.1g MnCl2·4H2O 0.03g

Na2MoO4·H2O 0.03g NiCl2·6H2O 0.02g CuCl2·2H2O 0.01g

Preparation of Trace Elements Solution SL-6: Add components

to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 3.4

Preparation of Medium: Add components, except Na2S·9H2O, NaCHO3,and L-cysteine·HCl·H2O solutions, to distilled/deionized wa-ter and bring volume to 1.0L Mix thoroughly Gently heat and bring to boiling Cool under 80% N2 + 20% CO2 Add L-cysteine·HCl·H2O and

Na2S·9H2O Add sufficient NaCHO3 solution to bring pH to 7.2 under 80% N2 + 20% CO2 Anaerobically distribute into tubes under 80% N2 + 20% CO2 Autoclave for 15 min at 15 psi pressure–121°C

Use: For the cultivation and maintenance of Clostridium xylanolyti-cum and other microorganisms that can utilize xylan as a carbon

source

Xylella Agar

(LMG Medium 115)

Agar 17.0g Yeast extract 10.0g ACES 10.0g Activated charcoal 2.0g α-ketoglutarate 1.0g

KOH, 1N 40mL

L-Cysteine-iron solution 20.0mL

pH 6.9 ± 0.2 at 25°C

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1922 Xylella fastidiosa Medium

L -Cysteine-Iron Solution:

L-Cysteine·HCl 0.4g

Fe4(P2O7)3 0.25g

Preparation of L -Cysteine-Iron Solution: Add components to

distilled/deionized water and bring volume to 20.0mL Mix

Preparation of Medium: Add ACES to 500.0mL of distilled water

at 50°C Combine with a solution containing 40.0mL of 1N KOH in

440.0mL of distilled water Add the other components except cysteine

iron solution Mix thoroughly Gently heat and bring to boiling

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

add 20.0mL sterile cysteine iron solution Pour into sterile Petri dishes

or distribute into sterile tubes

Use: For the cultivation of Xylella spp.

Xylella fastidiosa Medium

(LMG 115)

Agar 17.0g

Yeast extract 10.0g

ACES buffer 10.0g

Activated charcoal 2.0g

L-Cysteine-iron solution 20.0mL

pH 6.9 ± 0.2 at 25°C

L -Cysteine-Iron Solution:

L-Cysteine·HCl 0.4g

Fe4(P2O7)3 0.25g

Preparation of L -Cysteine-Iron Solution: Add components to

distilled/deionized water and bring volume to 20.0mL Mix

Preparation of Medium: Add ACES to 500.0mL of

distilled/de-ionized water at 50°C Add a solution containing 40.0mL of 1N KOH

in 440.0mL of distilled water Mix thoroughly Add the remaining

components, except L-cysteine-iron solution Mix thoroughly Gently

heat and bring to boiling Adjust pH to 6.9 with KOH Autoclave for

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

20.0mL of sterile L-cysteine-iron solution Mix thoroughly Pour into

sterile Petri dishes or distribute into sterile tubes

Use: For the cultivation of Xylella fastidiosa

Xylophilus Medium

CaCO3 20.0g

Agar 15.0g

D-Galactose 10.0g

Yeast extract 10.0g

Ferric ammonium citrate solution 10.0mL

Ferric Ammonium Citrate Solution:

Preparation of Ferric Ammonium Citrate Solution: Add

fer-ric ammonium citrate to distilled/deionized water and bring volume to

10.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–

121°C

Preparation of Medium: Add components, except ferric

ammoni-um citrate solution, to distilled/deionized water and bring volammoni-ume 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 ferric ammonium citrate solution Mix

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

Use: For the cultivation and maintenance of Xylophilus ampelina

Xylose Lactose Tergitol™ 4

(XLT-4)

Agar 18.0g Lactose 7.5g Sucrose 7.5g

Na2S2O3 6.8g Lysine 5.0g NaCl 5.0g Xylose 3.75g Yeast extract 3.0g Proteose peptone 1.6g Ferric ammonium citrate 0.8g Phenol Red 0.08g Selective supplement solution 4.6mL

pH 7.4 ± 0.2 at 25°C

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

Preparation of Selective Supplement Solution: Available as a premixed solution

Preparation of Medium: Add components, except selective sup-plement solution, to distilled/deionized water and bring volume to 1.0L Mix thoroughly Add 4.6mL of selective supplement solution Mix thoroughly Gently heat while stirring and bring to boiling Do not autoclave Cool to 50°C Mix thoroughly Pour into sterile Petri dishes

Use: For the isolation and identification of salmonellae from clinical, environmental, and food samples The presence of the selective agent, Tergitol™ 4, in this medium inhibits many organisms that can be prob-lematic on other plating media In addition, biochemical and pH

changes within the medium allow Salmonella spp (black colonies) to

be differentiated from organisms such as E coli (yellow colonies) and Shigella spp (red colonies) The enhanced selectivity of XLT-4 Agar

reduces the need for further identification procedures, saving time and money, and results in fewer false presumptive positive colonies when

compared to other Salmonella plating media.

Xylose Lysine Agar Base

See: XL Agar Base

Xylose Lysine Desoxycholate Agar

See: XLD Agar

Xylose Sodium Deoxycholate Citrate Agar

Agar 12.0g Xylose 10.0g Sodium citrate 5.0g

Na2S2O3·5H2O 5.0g

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Y 1 Adrenal Cell Growth Medium 1923

Beef extract 5.0g

Peptone 5.0g

NaCl 2.5g

Sodium deoxycholate 2.5g

Neutral Red (1% solution) 2.5mL

pH 7.5 ± 0.2 at 25°C

to boiling for 20 sec Do not autoclave Cool to 45°–50°C Pour into

sterile Petri dishes

Use: For the cultivation of Salmonella species and some Shigella species

Xylose YP Agar (Xylose Yeast Extract Peptone Agar)

CaCO3 20.0g

Agar 15.0g

Xylose 10.0g

Yeast extract 10.0g

Peptone 10.0g

MgSO4·7H2O 0.2g

MnSO4·4H2O 0.01g

FeSO4·7H2O 0.01g

NaCl 0.01g

pH 6.8 ± 0.2 at 25°C

to boiling Distribute into screw-capped test tubes Autoclave for 15

min at 15 psi pressure–121°C Adjust pH to 6.8 Mix thoroughly Pour

into sterile Petri dishes or leave in tubes

Use: For the cultivation and maintenance of Lactobacillus

vaccinos-tercus and other microorganisms that utilize xylose as a carbon source.

Xylose YP Broth (Xylose Yeast Extract Peptone Broth)

Xylose 10.0g

Yeast extract 10.0g

Peptone 10.0g

MgSO4·7H2O 0.2g

MnSO4·4H2O 0.01g

FeSO4·7H2O 0.01g

NaCl 0.01g

pH 6.8 ± 0.2 at 25°C

water and bring volume to 1.0L Mix thoroughly Distribute into

screw-capped test tubes Autoclave for 15 min at 15 psi pressure–121°C

Use: For the cultivation of Lactobacillus vaccinostercus and other

micro-organisms that utilize xylose as a carbon source

Y 1 Adrenal Cell Growth Medium

Ham’s F-10 medium 90.0mL

Fetal bovine serum 10.0mL

Penicillin-streptomycin solution 1.0mL

pH 7.0 ± 0.2 at 25°C

Ham’s F-10 Medium:

NaCl 7.4g NaHCO3 1.2g Glucose 1.1g NaH2PO4·H2O 0.29g KCl 0.28g L-Arginine·HCl 0.21g L-Glutamine 0.15g MgSO4·7H2O 0.15g Sodium pyruvate 0.11g

KH2PO4 0.08g CaCl2·2H2O 0.04g L-Cystine·2HCl 0.04g L-Histidine·HCl·H2O 0.02g L-Lysine·HCl 0.02g L-Asparagine-H2O 0.01g L-Aspartic Acid 0.01g L-Glutamic acid 0.01g L-Leucine 0.01g L-Proline 0.01g L-Serine 0.01g L-Alanine 8.9mg Glycine 7.5mg D-Phenylalanine 5.0mg L-Methionine 4.5mg Hypoxanthine 4.1mg L-Threonine 3.6mg L-Valine 3.5mg L-Isoleucine 2.6mg L-Tyrosine 1.8mg Vitamin B12 1.4mg Folic acid 1.3mg Phenol Red 1.2mg Thiamine·HCl 1.0mg FeSO4·7H2O 0.8mg Choline chloride 0.7mg D-Calcium pantothenate 0.7mg Thymidine 0.7mg Niacinamide 0.6mg L-Tryptophan 0.6mg Isoinositol 0.5mg Riboflavin 0.4mg Lipoic acid 0.2mg Pyridoxine·HCl 0.2mg ZnSO4·7H2O 0.03mg Biotin 0.02mg CuSO4·5H2O 3.0μg

Preparation of Ham’s F-10 Medium: Add components to dis-tilled/deionized water and bring volume to 1.0L Mix thoroughly

Penicillin-Streptomycin Solution:

Streptomycin 0.5g Penicillin G 500,000U

Preparation of Penicillin-Streptomycin Solution: Add compo-nents to distilled/deionized water and bring volume to 100.0mL Mix thoroughly Filter sterilize

Preparation of Medium: Aseptically combine components Filter sterilize Store at 4–5°C

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1924 Y 1 Adrenal Cell Growth Medium

Use: For the cultivation of Y-1 mouse adrenal tissue culture cells used

for the detection of heat-labile toxin (LT) produced by enterotoxigenic

strains of Escherichia coli LT causes the conversion of elongated

fibroblast-like cells into round, refractile cells

Y 1 Adrenal Cell Growth Medium

Ham’s F-10 medium 500.0mL

Fetal bovine serum 75.0mL

Penicillin-streptomycin solution 5.0mL

pH 7.0 ± 0.2 at 25°C

Ham’s F-10 Medium:

NaCl 7.4g

NaHCO3 1.2g

Glucose 1.1g

NaH2PO4·H2O 0.29g

KCl 0.28g

L-Arginine·HCl 0.21g

L-Glutamine 0.15g

MgSO4·7H2O 0.15g

Sodium pyruvate 0.11g

KH2PO4 0.08g

CaCl2·2H2O 0.04g

L-Cystine·2HCl 0.04g

L-Histidine·HCl·H2O 0.02g

L-Lysine·HCl 0.02g

L-Asparagine-H2O 0.01g

L-Aspartic Acid 0.01g

L-Glutamic acid 0.01g

L-Leucine 0.01g

L-Proline 0.01g

L-Serine 0.01g

L-Alanine 8.9mg

Glycine 7.5mg

D-Phenylalanine 5.0mg

L-Methionine 4.5mg

Hypoxanthine 4.1mg

L-Threonine 3.6mg

L-Valine 3.5mg

L-Isoleucine 2.6mg

L-Tyrosine 1.8mg

Vitamin B12 1.4mg

Folic acid 1.3mg

Phenol Red 1.2mg

Thiamine·HCl 1.0mg

FeSO4·7H2O 0.8mg

Choline chloride 0.7mg

D-Calcium pantothenate 0.7mg

Thymidine 0.7mg

Niacinamide 0.6mg

L-Tryptophan 0.6mg

Isoinositol 0.5mg

Riboflavin 0.4mg

Lipoic acid 0.2mg

Pyridoxine·HCl 0.2mg

ZnSO4·7H2O 0.03mg

Biotin 0.02mg

CuSO4·5H2O 3.0μg

Preparation of Ham’s F-10 Medium: Add components to

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

Penicillin-Streptomycin Solution:

Streptomycin 0.5g Penicillin G 500,000U

Preparation of Penicillin-Streptomycin Solution: Add compo-nents to distilled/deionized water and bring volume to 100.0mL Mix thoroughly Filter sterilize

Preparation of Medium: Aseptically combine components Filter sterilize Store at 4°–5°C

Use: For the cultivation of Y-1 mouse adrenal tissue culture cells used for the detection of cholera enterotoxin (CT) produced by enterotoxigenic

strains of Vibrio cholerae or Vibrio mimicus CT causes the conversion of

elongated fibroblast-like cells into round, refractile cells

YA12

Agar 10.0g Glucose 0.6g NaCl 0.5g Beef extract 0.2g Yeast extract 0.02g

pH 6.5–6.7 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 6.5–6.7 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 of Adelphamoeba galeacystis.

YA Halophile Medium

NaCl 100.0g Agar 15.0g Sodium acetate·3H2O 10.0g

Na2HPO4 3.8g

KH2PO4 1.3g Mg(NO3)2·6H2O 1.0g (NH4)2SO4 1.0g Yeast extract 1.0g

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components, except magnesium ni-trate, to tap water and bring volume to 1.0L Mix thoroughly Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C Aseptically add magnesium nitrate Adjust pH 7.2 with sterile KOH Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation and maintenance of halophilic

microorgan-isms, including Bacillus halodenitrificans.

YB Medium (Yeast Extract Beef Extract Medium)

Agar 20.0g Peptone 10.0g Beef extract 7.0g Yeast extract 5.0g NaCl 3.0g Thiourea 0.1g Methanol 20.0mL

pH 7.2 ± 0.2 at 25°C

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