Handbook of Microbiological Media, Fourth Edition part 193 pdf

Xanthobacter agilis Agar 1915 NH 4 Cl 1.0g K 2 HPO 4 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 Composition per liter: Malt extract 15.0g Maltose 12.75g Dextrin 2.75g NH 4 Cl 1.0g K 2 HPO 4 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. Use: For the cultivation and enumeration of yeasts. The low pH of the agar selectively inhibits bacterial growth. Wort Sucrose Agar Composition per liter: Agar 25.0g Yeast extract 1.0g Wort solution 500.0mL Sucrose solution 500.0mL Wort Solution: Composition per 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: Composition per 500.0mL: 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 Composition per liter: Yeast extract 1.0g Wort solution 500.0mL Sucrose solution 500.0mL Wort Solution: Composition per 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: Composition per 500.0mL: 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 Composition per liter: Solution 1 900.0mL Solution 2 100.0mL pH 7.0 ± 0.2 at 25°C Solution 1: Composition per 900.0mL: 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: Composition per 100.0mL: 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 Composition per 1100.0mL: Solution A 1.0L Solution B 100.0mL Solution A: Composition per liter: Agar 15.0g NaH 2 PO 4 ·12H 2 O 9.0g KH 2 PO 4 1.5g © 2010 by Taylor and Francis Group, LLC 1916 Xanthomonas Agar NH 4 ·Cl 1.0g Sodium propionate or 3-hydroxybutyrate 1.0g MgSO 4 ·7H 2 O 0.2g Trace elements solution 1.0mL pH 7.0 ± 0.2 at 25°C Trace Elements Solution: Composition per 2.0mL: H 3 BO 4 560.0μg ZnSO 4 ·7H 2 O 350.0μg NiCl 2 ·H 2 O 160.0μg Na 2 MoO 4 ·2H 2 O 100.0μg CuSO 4 ·5H 2 O 16.0μg MnCl 2 ·4H 2 O 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 water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Solution B: Composition per 100.0mL: Ferric ammonium citrate 50.0mg CaCl 2 ·2H 2 O 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 solution B. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation of Xanthobacter agilis. Xanthomonas Agar Composition per liter: Agar 15.0g 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 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 species. Xanthomonas Agar Composition per liter: CaCO 3 30.0g Agar 15.0g Glucose 10.0g Yeast extract 5.0g pH 7.0 ± 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. Cool rapidly. Use: For the cultivation and maintenance of Alcaligenes latus, Erwinia tracheiphila, Pseudomonas amygdali, Xanthomonas albilineans, Xan- thomonas axonopodis, Xanthomonas campestris, Xanthomonas fragariae, Xanthomonas maltophilia, Xanthomonas oryzae, and Xylophilus ampeli- nus. Xanthomonas albilineans Agar Composition per liter: 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 Composition per liter: Trizma ® (tris[hydroxymethyl] aminomethane) base 6.04g Glucose 5.0g KCl 1.0g NaCl 1.0g L-Phenylalanine 0.9g MgSO 4 0.2g L-Arginine 0.1g L-Methionine 0.1g (NH 4 ) 2 SO 4 0.1g NH 4 Cl 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 sterilize. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation of Stenotrophomonas maltophilia. Xanthomonas Medium Composition per liter: 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 © 2010 by Taylor and Francis Group, LLC XED-AGAR 1917 Glucose 5.0g Yeast extract 3.0g K 2 HPO 4 0.7g MgSO 4 ·7H 2 O 0.25g 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 Xanthomonas species. XB45/XB90/PB90-2 Medium (DSMZ Medium 862) Composition per 1069.2mL: 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: Composition per 940.0mL: NaCl 1.0g KCl 0.5g MgCl 2 ·6H 2 O 0.4g KH 2 PO 4 0.2g NH 4 Cl 0.25g CaCl 2 ·2H 2 O 0.15g Resazurin 0.5mg Preparation of Solution A: Prepare under 80% N 2 + 20% CO 2 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% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution B: Composition per liter: FeCl 2 ·4H 2 O 1.5g CoCl 2 ·6H 2 O 190.0mg MnCl 2 ·4H 2 O 100.0mg ZnCl 2 70.0mg Na 2 MoO 4 ·2H 2 O 36.0mg NiCl 2 ·6H 2 O 24.0mg H 3 BO 3 6.0mg CuCl 2 ·2H 2 O 2.0mg HCl (25% solution) 10.0mL Preparation of Solution B: Add FeCl 2 ·4H 2 O 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% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution C: Composition per liter: Pyridoxine hydrochloride 300.0mg Thiamine-HCl·2H 2 O 200.0mg Nicotinic acid 200.0mg Vitamin B 12 100.0mg 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% N 2 . Mix thorough- ly. Filter sterilize. Solution D: Composition per liter: Pyridoxine-HCl 10.0mg Thiamine-HCl·2H 2 O 5.0mg Riboflavin 5.0mg Nicotinic acid 5.0mg D-Ca-pantothenate 5.0mg p-Aminobenzoic acid 5.0mg Lipoic acid 5.0mg Biotin 2.0mg Folic acid 2.0mg Vitamin B 12 0.1mg Preparation of Solution D: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize. Solution E: Composition per 100.0mL: NaHCO 3 5.0g Preparation of Solution E: Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution F: Composition per 10.0mL: Glucose 1.0g Preparation of Solution F: Add glucose to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Auto- clave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution G: Composition per 10.0mL: Na 2 S·9H 2 O 0.125g Preparation of Solution G: Add Na 2 S·9H 2 O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool to 25°C. Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 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 solution G to 940.0mL solution A. Distribute an- aerobically under 80% N 2 + 20% CO 2 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) Composition per liter: Agar 18.0g Xylan 7.0g Yeast extract 3.0g pH 7.0 ± 0.2 at 25°C © 2010 by Taylor and Francis Group, LLC 1918 Xenorhabdus Agar Preparation of Medium: Add components to distilled/deionized 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 Composition per liter: Agar 15.0g Peptone 10.0g NaCl 5.0g Yeast extract 5.0g 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. 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 Composition per liter: Peptone 10.0g NaCl 5.0g Yeast extract 5.0g pH 7.2 ± 0.2 at 25°C Preparation of Medium: Add components to distilled/deionized wa- ter and bring volume to 1.0L. Mix thoroughly. 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) Composition per liter: 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: Composition per 100.0mL: Na 2 S 2 O 3 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 Composition per liter: 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: Composition per 100.0mL: Na 2 S 2 O 3 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) Composition per liter: Agar 13.5g Lactose 7.5g Sucrose 7.5g Na 2 S 2 O 3 6.8g L-Lysine 5.0g NaCl 5.0g Xylose 3.5g © 2010 by Taylor and Francis Group, LLC XLT4 HiVeg Agar Base 1919 Yeast extract 3.0g Sodium desoxycholate 2.5g Ferric ammonium citrate 0.8g Phenol Red 0.08g pH 7.5 ± 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 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. 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) Composition per liter: Agar 15.0g Lactose 7.5g Sucrose 7.5g Na 2 S 2 O 3 6.8g L-Lysine 5.0g NaCl 5.0g Xylose 3.75g Yeast extract 3.0g Sodium deoxycholate 2.5g Ferric ammonium citrate 0.8g Phenol Red 0.08g pH 7.5 ± 0.2 at 25°C Source: This medium is available as a premixed powder from BD Di- agnostic Systems and Oxoid. 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. 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-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) Composition per liter: Agar 15.0g Lactose 7.5g Sucrose 7.5g Na 2 S 2 O 3 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: Composition per 100.0mL: 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. 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-fer- menting, lysine-nondecarboxylating bacteria appear as opaque yellow colonies. Lactose- or sucrose-fermenting bacteria appear as yellow col- onies. XLT4 HiVeg Agar Base Composition per liter: Agar 18.0g Lactose 7.5g Saccharose 7.5g Na 2 S 2 O 3 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. Selective Supplement Solution: Composition per 100.0mL: 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. 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. © 2010 by Taylor and Francis Group, LLC 1920 XPS Agar Use: For the isolation and differentiation of enteric pathogens, espe- cially Shigella and Providencia species. XPS Agar Composition per liter: 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 Na 2 HPO 4 0.79g Ca(NO 3 ) 2 ·4H 2 O 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: Composition per 10.0mL: Ca(NO 3 ) 2 ·4H 2 O 0.5g Preparation of Ca(NO 3 ) 2 ·4H 2 O Solution: Add Ca(NO 3 ) 2 ·4H 2 O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Solution A: Add components, except Ca(NO 3 ) 2 ·4H 2 O 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(NO 3 ) 2 ·4H 2 O 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 solution B. Mix thoroughly. Aseptically pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation of Xanthomonas campestris. XPS Broth Composition per liter: Potatoes, infusion from 40.0g Sucrose 15.0g Peptone 5.0g Glucose 4.0g Casamino acids 1.0g Na 2 HPO 4 0.79g Ca(NO 3 ) 2 ·4H 2 O solution 10.0mL pH 5.1 ± 0.2 at 25°C Potatoes, Infusion From: Composition per 500.0mL: 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 ·4H 2 O Solution: Composition per 10.0mL: Ca(NO 3 ) 2 ·4H 2 O 0.5g Preparation of Ca(NO 3 ) 2 ·4H 2 O Solution: Add Ca(NO 3 ) 2 ·4H 2 O to distilled/deionized water and bring volume to 10.0mL. Mix thor- oughly. Filter sterilize. Preparation of Medium: Add components, except Ca(NO 3 ) 2 ·4H 2 O 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(NO 3 ) 2 ·4H 2 O 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) Composition per liter: Potatoes, infusion from 40.0g Sucrose 15.0g Peptone 5.0g Glucose 4.0g Casamino acids 1.0g Na 2 HPO 4 0.79g Thymidine 10.0mg Ca(NO 3 ) 2 ·4H 2 O solution 10.0mL pH 5.1 ± 0.2 at 25°C Potatoes, Infusion From: Composition per 500.0mL: 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 ·4H 2 O Solution: Composition per 10.0mL: Ca(NO 3 ) 2 ·4H 2 O 0.5g Preparation of Ca(NO 3 ) 2 ·4H 2 O Solution: Add Ca(NO 3 ) 2 ·4H 2 O to distilled/deionized water and bring volume to 10.0mL. Mix thor- oughly. Filter sterilize. Preparation of Medium: Add components, except Ca(NO 3 ) 2 ·4H 2 O 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(NO 3 ) 2 ·4H 2 O solution. Mix thorough- ly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation of Xanthomonas oryzae. XSM Agar Composition per liter: Agar 15.0g Glucose 5.0g Sucrose 2.0g Malt extract 1.0g © 2010 by Taylor and Francis Group, LLC 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 Composition per liter: Xylan 30.0g Agar 12.0g Peptone 2.0g Yeast extract 0.5g L-Cysteine·HCl·H 2 O 0.25g Na 2 S·9H 2 O 0.25g Rumen fluid 400.0mL NaHCO 3 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: Composition per 100.0mL: NaHCO 3 3.96g Preparation of NaHCO 3 Solution: Add NaHCO 3 to distilled/de- ionized water and bring volume to 100.0mL. Mix thoroughly. Gas with 100% CO 2 . Mineral Solution I: Composition per liter: K 2 HPO 4 3.0g Preparation of Mineral Solution I: Add K 2 HPO 4 to distilled/de- ionized water and bring volume to 1.0L. Mix thoroughly. Mineral Solution II: Composition per liter: Sodium citrate 20.0g NaCl 12.0g KH 2 PO 4 6.0g MgCl 2 ·6H 2 O 2.0g CaCl 2 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: Composition per liter: Pyridoxine·HCl 10.0mg Thiamine·HCl 5.0mg Riboflavin 5.0mg Nicotinic acid 5.0mg Calcium pantothenate 5.0mg p-Aminobenzoic acid 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: Composition per liter: 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: Composition per 100.0mL: Hemin 0.01g Preparation of Hemin Solution: Add hemin to 100.0mL of 0.01N NaOH. Mix thoroughly. Trace Elements Solution SL-6: Composition per liter: H 3 BO 3 0.3g CoCl 2 ·6H 2 O 0.2g ZnSO 4 ·7H 2 O 0.1g MnCl 2 ·4H 2 O 0.03g Na 2 MoO 4 ·H 2 O 0.03g NiCl 2 ·6H 2 O 0.02g CuCl 2· 2H 2 O 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 Na 2 S·9H 2 O, NaCHO 3 ,and L-cysteine·HCl·H 2 O solutions, to distilled/deionized wa- ter and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Cool under 80% N 2 + 20% CO 2 . Add L-cysteine·HCl·H 2 O and Na 2 S·9H 2 O. Add sufficient NaCHO 3 solution to bring pH to 7.2 under 80% N 2 + 20% CO 2 . Anaerobically distribute into tubes under 80% N 2 + 20% CO 2 . 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) Composition per liter: 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 © 2010 by Taylor and Francis Group, LLC 1922 Xylella fastidiosa Medium L-Cysteine-Iron Solution: Composition per 20.0mL: L-Cysteine·HCl 0.4g Fe 4 (P 2 O 7 ) 3 0.25g Preparation of L-Cysteine-Iron Solution: Add components to distilled/deionized water and bring volume to 20.0mL. Mix thorough- ly. Filter sterilize. 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) Composition per liter: 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: Composition per 20.0mL: L-Cysteine·HCl 0.4g Fe 4 (P 2 O 7 ) 3 0.25g Preparation of L-Cysteine-Iron Solution: Add components to distilled/deionized water and bring volume to 20.0mL. Mix thorough- ly. Filter sterilize. 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 Composition per liter: CaCO 3 20.0g Agar 15.0g D-Galactose 10.0g Yeast extract 10.0g Ferric ammonium citrate solution 10.0mL Ferric Ammonium Citrate Solution: Composition per 10.0mL: Ferric ammonium citrate 0.25g 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 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 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) Composition per 1004.6mL: Agar 18.0g Lactose 7.5g Sucrose 7.5g Na 2 S 2 O 3 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. Selective Supplement Solution: Composition per 100.0mL: Tergitol™ 4 Proprietary 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 Composition per liter: Agar 12.0g Xylose 10.0g Sodium citrate 5.0g Na 2 S 2 O 3 ·5H 2 O 5.0g © 2010 by Taylor and Francis Group, LLC Y 1 Adrenal Cell Growth Medium 1923 Beef extract 5.0g Peptone 5.0g NaCl 2.5g Sodium deoxycholate 2.5g Ferric ammonium citrate 1.0g Neutral Red (1% solution) 2.5mL pH 7.5 ± 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 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) Composition per liter: CaCO 3 20.0g Agar 15.0g Xylose 10.0g Yeast extract 10.0g Peptone 10.0g MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 0.01g FeSO 4 ·7H 2 O 0.01g NaCl 0.01g 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 and bring 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) Composition per liter: Xylose 10.0g Yeast extract 10.0g Peptone 10.0g MgSO 4 ·7H 2 O 0.2g MnSO 4 ·4H 2 O 0.01g FeSO 4 ·7H 2 O 0.01g NaCl 0.01g 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. 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 Composition per 101.0mL: 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: Composition per liter: NaCl 7.4g NaHCO 3 1.2g Glucose 1.1g NaH 2 PO 4 ·H 2 O 0.29g KCl 0.28g L-Arginine·HCl 0.21g L-Glutamine 0.15g MgSO 4 ·7H 2 O 0.15g Sodium pyruvate 0.11g KH 2 PO 4 0.08g CaCl 2 ·2H 2 O 0.04g L-Cystine·2HCl 0.04g L-Histidine·HCl·H 2 O 0.02g L-Lysine·HCl 0.02g L-Asparagine-H 2 O 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 B 12 1.4mg Folic acid 1.3mg Phenol Red 1.2mg Thiamine·HCl 1.0mg FeSO 4 ·7H 2 O 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 ZnSO 4 ·7H 2 O 0.03mg Biotin 0.02mg CuSO 4 ·5H 2 O 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: Composition per 100.0mL: 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. © 2010 by Taylor and Francis Group, LLC 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 Composition per 580.0mL: 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: Composition per liter: NaCl 7.4g NaHCO 3 1.2g Glucose 1.1g NaH 2 PO 4 ·H 2 O 0.29g KCl 0.28g L-Arginine·HCl 0.21g L-Glutamine 0.15g MgSO 4 ·7H 2 O 0.15g Sodium pyruvate 0.11g KH 2 PO 4 0.08g CaCl 2 ·2H 2 O 0.04g L-Cystine·2HCl 0.04g L-Histidine·HCl·H 2 O 0.02g L-Lysine·HCl 0.02g L-Asparagine-H 2 O 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 B 12 1.4mg Folic acid 1.3mg Phenol Red 1.2mg Thiamine·HCl 1.0mg FeSO 4 ·7H 2 O 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 ZnSO 4 ·7H 2 O 0.03mg Biotin 0.02mg CuSO 4 ·5H 2 O 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: Composition per 100.0mL: 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 Composition per liter: 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 Composition per liter: NaCl 100.0g Agar 15.0g Sodium acetate·3H 2 O 10.0g Na 2 HPO 4 3.8g KH 2 PO 4 1.3g Mg(NO 3 ) 2 ·6H 2 O 1.0g (NH 4 ) 2 SO 4 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) Composition per liter: 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 © 2010 by Taylor and Francis Group, LLC . 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. 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. cultivation of Stenotrophomonas maltophilia. Xanthomonas Medium Composition per liter: Pancreatic digest of gelatin 10.0g Sucrose 10.0g Beef extract 6.0g pH 6.8 ± 0.2 at 25°C Preparation of Medium: