Handbook of Microbiological Media, Fourth Edition part 62 docx

Double Sugar Agar, Russell 605 Dorset Egg Medium Composition per liter: Homogenized whole egg 950.0mL Glycerol 50.0mL pH 6.8–7.4 at 25°C Source: This medium is available as a prepared medium from BD Di- agnostic Systems. Homogenized Whole Egg: Composition per liter: Whole eggs 18–24 Preparation of Homogenized Whole Egg: Use fresh eggs, less than 1 week old. Scrub the shells with soap. Let stand in a soap solution for 30 min. Rinse in running water. Soak eggs in 70% ethanol for 15 min. Break the eggs into a sterile container. Homogenize by shaking. Filter through four layers of sterile cheesecloth into a sterile graduated cylinder. Measure out 1.0L. Preparation of Medium: Filter sterilize glycerol. Combine glycerol and homogenized whole egg. Mix thoroughly. Distribute into sterile screw-capped tubes. Place tubes in a slanted position. Inspissate at 85°C (moist heat) for 45 min. Use: For the maintenance of Mycobacterium species. Double-Strength Crude Lactobacillus Medium Composition per 1025.0mL: Yeast extract (Basamine) 20.0g Sucrose 20.0g Casein hydrolysate 15.0g Potassium acetate 3.0g Histidine·HCl·H 2 O 2.0g Ascorbic acid 1.0g Pyridoxamine·HCl 33.0μg Salts solution A 20.0mL Salts solution B 5.0mL pH 5.4 ± 0.2 at 25°C Salts Solution A: Composition per liter: K 2 HPO 4 ·3H 2 O 16.5g KH 2 PO 4 ·H 2 O 16.5g Preparation of Salts Solution A: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Salts Solution B: Composition per liter: MgSO 4 ·7H 2 O 8.0g FeSO 4 ·7H 2 O 0.4g NaCl 0.4g HCl, concentrated 0.1mL MnSO 4 ·H 2 O 0.1mL Preparation of Salts Solution B: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: Add components, except salts solution A and salts solution B, to distilled/deionized water and bring volume to 975.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure– 121°C. Aseptically add 20.0mL of sterile salts solution A and 5.0mL of sterile salts solution B. Mix thoroughly. Adjust pH to 5.4. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation of Lactobacillus species. Double-Strength Crude Medium for Lactobacillus Composition per liter: Sucrose 20.0g Yeast extract 20.0g Casein hydrolysate 15.0g Potassium acetate 3.0g Histidine·HCl 2.0g Ascorbic acid 1.0g Pyridoxamine·HCl 33.0μg Salts solution A 20.0mL Salts solution B 5.0mL pH 5.4 ± 0.2 at 25°C Salts Solution A: Composition per 100.0mL: KH 2 PO 4 ·H 2 O 16.5g K 2 HPO 4 ·3H 2 O 16.5g Preparation of Salts Solution A: Add components to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Salts Solution B: Composition per 100.0mL: MgSO 4 ·7H 2 O 8.0g FeSO 4 ·7H 2 O 0.4g MnSO 4 ·H2O 0.4g NaCl 0.4g HCl, concentrated 0.1mL Preparation of Salts Solution B: Add components to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. 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 5.4 with acetic acid. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation and maintenance of Lactobacillus species. Double Sugar Agar, Russell (Russell Double Sugar Agar) Composition per liter: Agar 15.0g Lactose 10.0g Casein enzymic hydrolysate 7.5g NaCl 5.0g Beef extract 3.0g Peptic digest of animal tissue 2.5g Glucose 1.0g Phenol Red 0.025g pH 7.3 ± 0.2 at 25°C Source: This medium is available from HiMedia. 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. Allow the tubes to solidify in slanting position to form a generous butt. Use: For the differentiation of Gram-negative enteric bacilli on the basis of their ability to ferment glucose and lactose with or without gas formation. © 2010 by Taylor and Francis Group, LLC 606 Double Sugar HiVeg Agar Double Sugar HiVeg Agar Composition per liter: Agar 15.0g Lactose 10.0g Plant hydrolysate 7.5g NaCl 5.0g Plant extract 3.0g Plant peptone 2.5g Glucose 1.0g Phenol Red 0.025g pH 7.3 ± 0.2 at 25°C Source: This medium is available as a premixed powder from Hi- Media. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes. Autoclave for 15 min at 15 psi pressure– 121°C. Allow tubes to cool in a slanted position. Use: For the identification of Gram-negative enteric bacilli based on their fermentation of glucose and lactose. Bacteria that ferment both glucose and lactose produce a yellow slant and yellow butt. Bacteria that ferment glucose but do not ferment lactose produce a red slant and a yellow butt. Bacteria that ferment neither glucose nor lactose produce an unchanged pink-orange color. Doyle and Roman Enrichment Medium Composition per liter: Pancreatic digest of casein 10.0g Peptic digest of animal tissue 10.0g NaCl 5.0g Sodium succinate 3.0g Yeast extract 2.0g Glucose 1.0g NaHSO 3 0.1g L-Cysteine·HCl·H 2 O 0.1g Horse blood, lysed 70.0mL Antibiotic solution 10.0mL pH 7.0 ± 0.2 at 25°C Antibiotic Solution: Composition per 10.0mL: Cycloheximide 0.05g Vancomycin 0.015g Trimethoprim lactate 5.0mg Polymyxin B 200,000U Preparation of Antibiotic Solution: Add components to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Caution: Cycloheximide is toxic. Avoid skin contact or aerosol formation and inhalation. Preparation of Medium: Add components, except antibiotic solution and horse blood, to distilled/deionized water and bring volume to 920.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add sterile antibiotic solution and horse blood. Mix thoroughly. Asep- tically distribute into sterile flasks in 90.0–100.0mL volumes. Use: For the cultivation and enrichment of Campylobacter species from foods. Doyle's Enrichment Broth Base with Antibiotic Solution Composition per liter: Casein enzymatic hydrolysate 10.0g Peptic digest of animal tissue 10.0g NaCl 5.0g Sodium succinate 3.0g Yeast extract 2.0g Glucose 1.0g L-Cysteine·HCl 0.1g NaHSO 3 0.1g Antibiotic solution 10.0mL pH 7.0 ± 0.2 at 25°C Source: This medium, without antibiotic solution, is available as a premixed powder from HiMedia. Antibiotic Solution: Composition per 10.0mL: Cycloheximide 0.05g Vancomycin 0.015g Trimethoprim lactate 5.0mg Polymyxin B 200,000U Preparation of Antibiotic Solution: Add components to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Caution: Cycloheximide is toxic. Avoid skin contact or aerosol formation and inhalation. Preparation of Medium: Add components, except antibiotic 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 45°–50°C. Aseptically add a selective sterile antibiotic solution. Generally 50.0mL of defibrinated horse blood is also added as an enrichment. Mix thoroughly. Use: For the cultivation and enrichment of Campylobacter species from foods. Doyle's Enrichment HiVeg Broth Base with Antibiotic Solution Composition per liter: Plant hydrolysate 10.0g Plant peptone 10.0g NaCl 5.0g Sodium succinate 3.0g Yeast extract 2.0g Glucose 1.0g L-Cysteine·HCl 0.1g NaHSO 3 0.1g Antibiotic solution 10.0mL pH 7.0 ± 0.2 at 25°C Source: This medium, without antibiotic solution, is available as a premixed powder from HiMedia. Antibiotic Solution: Composition per 10.0mL: Cycloheximide 0.05g Vancomycin 0.015g Trimethoprim lactate 5.0mg Polymyxin B 200,000U © 2010 by Taylor and Francis Group, LLC Drigalski Litmus Lactose Agar 607 Preparation of Antibiotic Solution: Add components to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Caution: Cycloheximide is toxic. Avoid skin contact or aerosol formation and inhalation. Preparation of Medium: Add components, except antibiotic 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 45°–50°C. Aseptically add a selective sterile antibiotic solution. Generally 50.0mL of defibrinated horse blood is also added as an enrichment. Mix thoroughly. Use: For the cultivation and enrichment of Campylobacter species from foods. DP Agar Composition per liter: Agar 20.0g Pancreatic digest of casein 10.0g Yeast extract 10.0g K 2 HPO 4 7.0g Glucose 5.0g KH 2 PO 4 3.0g Trisodium citrate·3H 2 O 0.5g MgSO 4 ·7H 2 O 0.1g Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation of Escherichia coli. DPA with Calcium Carbonate Composition per liter: Agar 20.0g Glucose 20.0g Peptone 10.0g CaCO 3 5.0g Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 30 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Dekkera species. DPM Medium (DSMZ Medium 737) Composition per liter: Na-propionate 1.20g KH 2 PO 4 1.0g MgSO 4 ·7H 2 O 0.1g CaCl 2 ·2H 2 O 0.01g Chelated iron solution 1.8mL Trace elements solution 1.0mL pH 6.8 ± 0.2 at 25°C Chelated Iron Solution: Composition per liter: Na 2 -EDTA 7.56g FeSO 4 ·5H 2 O 5.54g Preparation of Chelated Iron Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Trace Elements Solution: Composition per liter: H 3 BO 3 2.860 MnCl 2 ·4H 2 O 1.81g ZnSO 4 ·7H 2 O 0.22g CuSO 4 ·5H 2 O 0.08g Na 2 MoO 4 ·4H 2 O 0.025g CoCl 2 0.025g Preparation of Trace Elements Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. 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 Frankia sp. DRBC Agar (Dichloran Rose Bengal Chloramphenicol Agar) Composition per liter: Agar 15.0g Glucose 10.0g Peptone 5.0g KH 2 PO 4 1.0g MgSO 4 ·7H 2 O 0.5g Rose Bengal 0.025g Dichloran 0.002g Chloramphenicol solution 10.0mL pH 5.6 ± 0.2 at 25°C Source: This medium is available as a premixed powder from Oxoid Unipath. Chloramphenicol Solution: Composition per 10.0mL: Chloramphenicol 0.1g Preparation of Chloramphenicol Solution: Add chloramphenicol to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except chloramphenicol 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 45°–50°C. Aseptically add sterile chloramphenicol solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the isolation, cultivation, and enumeration of yeasts and molds associated with food spoilage. Drigalski Litmus Lactose Agar Composition per liter: Agar 10.0g Lactose 10.0g Meat peptone 5.0g Beef extract 3.0g Litmus 1.0g pH 7.4 ± 0.2 at 25°C Source: This medium is available as a premixed powder from Sigma Aldrich. © 2010 by Taylor and Francis Group, LLC 608 Drigalski Litmus Lactose Agar 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 maintenance of lactic acid bacteria and differentiation of bacteria on the basis of lactose fermentation. Bacteria that ferment lactose appear as red colonies and others as dark blue-purple colonies. Drigalski Litmus Lactose Agar Composition per liter: Lactose 15.0g Agar 13.0g Peptic digest of animal tissue 7.0g NaCl 5.0g Litmus 1.2g pH 7.4 ± 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. 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 maintenance of lactic acid bacteria and differentiation of bacteria on the basis of lactose fermentation. Bacteria that ferment lactose appear as red colonies and others as dark blue-purple colonies. Drigalski Litmus Lactose Agar, Modified Composition per liter: Agar 16.0g Lactose 10.0g Peptic digest of animal tissue 10.0g Beef extract 4.0g Bromthymol Blue 0.04g pH 7.4 ± 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. 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 maintenance of lactic acid bacteria and differentiation of bacteria on the basis of lactose fermentation. Bacteria that ferment lactose appear as red colonies and others as dark blue-purple colonies. Drigalski Litmus Lactose HiVeg Agar Composition per liter: Lactose 15.0g Agar 13.0g Plant peptone 7.0g NaCl 5.0g Litmus 1.2g pH 7.4 ± 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. 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 maintenance of lactic acid bacteria and differentiation of bacteria on the basis of lactose fermentation. Bacteria that ferment lactose appear as red colonies and others as dark blue-purple colonies. DS Sporulation Medium, Modified See: Duncan-Strong Sporulation Medium, Modified DSA See: Dextrose Soil Agar DSA Cellulose See: Dextrose Sucrose Cellulose Agar DSIC Medium, Modified (DSMZ Medium 747) Composition per 994.0mL: Solution A 910.0mL Solution B 70.0mL Solution C 14.0mL pH 7.0–7.1 at 25°C Solution A: Composition per 960.0mL: NaCl 125.0g K 2 SO 4 2.5g Na-acetate 2.0g Yeast extract 0.75g KH 2 PO 4 0.6g NH 4 Cl 0.5g Na 2 S2O 3 ·5H 2 O 0.1g MOPS buffer 10.0mL Vitamin B 12 solution 1.0mL Trace elements solution SL-10 1.0mL Vitamin B 12 Solution: Composition per 100.0mL: Vitamin B 12 10.0mg Vitamin B 12 Solution: Add vitamin B 12 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge under 100% N 2 gas for 3 min. Trace Elements Solution SL-10: 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 Trace Elements Solution SL-10: Add FeCl 2 ·4H 2 O to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized water and bring volume 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. © 2010 by Taylor and Francis Group, LLC Dubos Broth 609 MOPS Buffer: Composition per 10.0mL: MOPS [3-(N-morpholino) propane sulfonic acid] 2.1g Na-acetate 0.3g EDTA 0.1g Preparation of MOPS Buffer: Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Adjust to pH 7.2. Filter sterilize. Preparation of Solution A: Add components to distilled/deionized water and bring volume to 960.0mL. Mix thoroughly. Sparge with 100% N 2 . Gently heat and bring to boiling while continuing to sparge with 100% N 2 . Distribute about 13mL aliquots in 15mL Hungate tubes. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution B: Composition per 70.0mL: MgCl 2 ·6H 2 O 10.0g CaCl 2 ·2H 2 O 0.2g Preparation of Solution B: Add components to distilled/deionized water and bring volume to 70.0mL. Mix thoroughly. Adjust pH to 7.0. Sparge with 100% N 2 . Gently heat and bring to boiling while continuing to sparge with 100% N 2 . Distribute into a screw-capped bottle. Auto- clave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Solution C: Composition per 14.0mL: NaHCO 3 1.0g Preparation of Solution C: Add NaHCO 3 to distilled/deionized water and bring volume to 14.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Filter sterilize. Preparation of Medium: Inject 1.0 ml of solution B and 0.2 ml of solution C in each tube of solution A. Use: For the cultivation of Rhodovibrio sodomensis=Rhodospirillum sodomense. DSM 134 See: Haliscomenobacter Medium DST Agar See: Diagnostic Sensitivity Test Agar DTC Agar Composition per liter: Agar 20.0g Pancreatic digest of casein 15.0g NaCl 5.0g Papaic digest of soybean meal 5.0g Deoxyribonucleic acid 2.0g Toluidine Blue O 0.1g Cephalothin solution 10.0mL Cephalothin Solution: Composition per 10.0mL: Cephalothin 1.0g Preparation of Cephalothin Solution: Add cephalothin to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except cephalothin 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 45°–50°C. Aseptically add sterile cephalothin solution. Mix thoroughly. Pour into sterile Petri dishes. Use: For the isolation and cultivation of Serratia species. Serratia appear as colonies with red halos. DTM Agar (Dermatophyte Test Medium Agar) Composition per liter: Agar 20.0g Enzymatic digest of soybean meal 10.0g Glucose 10.0g Cycloheximide 0.5g Phenol Red 0.2g Chlortetracycline 0.1g Gentamicin 0.1g pH 7.3 ± 0.2 at 25°C Source: Available as a prepared medium from BD Diagnostic Sys- tems. Caution: Cycloheximide is toxic. Avoid skin contact or aerosol formation and inhalation. 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 isolation and cultivation of dermatophytic fungi. Dubos Agar with Filter Paper Composition per liter: Agar 15.0g K 2 HPO 4 1.0g KCl 0.5g MgSO 4 ·7H 2 O 0.5g NaNO 3 0.5g FeSO 4 ·7H 2 O 0.01g 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. Autoclave for 15 min at 15 psi pressure– 121°C. Pour into sterile Petri dishes. Lay sterile strips of Whatman #1 filter paper on the surface of the agar. Use: For the cultivation and maintenance of Cytophaga hutchinsonii. Dubos Broth Composition per liter: Na 2 HPO 4 2.5g L-Asparagine 2.0g KH 2 PO 4 1.0g Pancreatic digest of casein 0.5g Tween™ 80 0.2g CaCl 2 ·2H 2 O 0.5mg CuSO 4 0.1mg ZnSO 4 ·7H 2 O 0.1mg Ferric ammonium citrate 0.05g MgSO 4 ·7H 2 O 0.01g Bovine serum albumin or bovine serum 20.0mL pH 6.5 ± 0.2 at 25°C © 2010 by Taylor and Francis Group, LLC 610 Dubos Broth Base with Serum and Glycerol Source: This medium is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components, except bovine serum or bovine serum albumin, to distilled/deionized water and bring volume to 980.0mL. Mix thoroughly. Gently heat and bring to boiling. Auto- clave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Asepti- cally add sterile bovine serum or bovine serum albumin. Mix thoroughly. Aseptically distribute into sterile tubes. Use: For the cultivation of Mycobacterium tuberculosis and other Mycobacterium species. Dubos Broth Base with Serum and Glycerol Composition per liter: Na 2 HPO 4 2.5g L-Asparagine 2.0g KH 2 PO 4 1.0g Casein enzymatic hydrolysate 0.5g Polysorbate 80 0.2g Ferric ammonium citrate 0.05g MgSO 4 0.01g CaCl 2 0.5mg CuSO 4 0.1mg ZnSO 4 0.1mg Glycerol 50.0mL Bovine serum or bovine albumin V 20.0mL pH 6.5 ± 0.2 at 25°C Source: This medium, without bovine serum or glycerol, is available as a premixed powder from HiMedia. Preparation of Medium: Add components, except bovine serum, 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 45°–50°C. Aseptically add 20.0mL sterile bovine serum or bovine serum albumin. Mix thoroughly. Aseptically distribute into sterile tubes. Use: For the cultivation of Mycobacterium tuberculosis and other Mycobacterium species. Dubos Broth with Horse Serum Composition per liter: Na 2 HPO 4 2.5g L-Asparagine 2.0g KH 2 PO 4 1.0g Pancreatic digest of casein 0.5g Tween™ 80 0.2g CaCl 2 ·2H 2 O 0.5mg CuSO 4 0.1mg ZnSO 4 ·7H 2 O 0.1mg Ferric ammonium citrate 0.05g MgSO 4 ·7H 2 O 0.01g Horse serum 50.0mL pH 6.5 ± 0.2 at 25°C Preparation of Medium: Add components, except horse serum, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add sterile horse serum. Mix thoroughly. Aseptically distribute into sterile tubes. Use: For the cultivation and maintenance of Corynebacterium species. Dubos HiVeg Broth Base with Serum and Glycerol Composition per liter: Na 2 HPO 4 2.5g L-Asparagine 2.0g KH 2 PO 4 1.0g Plant hydrolysate 0.5g Polysorbate 80 0.2g Ferric ammonium citrate 0.05g MgSO 4 0.01g CaCl 2 0.5mg CuSO 4 0.1mg ZnSO 4 0.1mg Glycerol 50.0mL Bovine serum or bovine albumin V 20.0mL pH 6.5 ± 0.2 at 25°C Source: This medium, without bovine serum or glycerol, is available as a premixed powder from HiMedia. Preparation of Medium: Add components, except bovine serum, 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 45°–50°C. Aseptically add 20.0mL sterile bovine serum or bovine serum albumin. Mix thoroughly. Aseptically distribute into sterile tubes. Use: For the cultivation of Mycobacterium tuberculosis and other Mycobacterium species. Dubos Mineral Medium Composition per liter: K 2 HPO 4 1.0g KCl 0.5g MgSO 4 ·7H 2 O 0.5g NaNO 3 0.5g FeSO 4 ·7H 2 O 0.01g 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 isolation and cultivation of Cytophaga species, Herpeto- siphon species, Saprospira species, and Flexithrix species. Dubos Oleic Agar Composition per liter: Agar 15.0g Na 2 HPO 4 2.5g KH 2 PO 4 1.0g L-Asparagine 1.0g Pancreatic digest of casein 0.5g Ferric ammonium citrate 0.05g MgSO 4 ·7H 2 O 0.01g CaCl 2 ·2H 2 O 0.5mg CuSO 4 0.1mg ZnSO 4 ·7H 2 O 0.1mg Dubos oleic albumin complex 20.0mL Penicillin solution 10.0mL pH 6.6 ± 0.2 at 25°C Source: This medium is available as a premixed powder from BD Di- agnostic Systems. © 2010 by Taylor and Francis Group, LLC Dubos Salts Agar with 1% Sodium Chloride 611 Dubos Oleic Albumin Complex: Composition per 100.0mL: Bovine serum albumin, fraction V 5.0g Oleic acid, sodium salt 0.05g NaCl (0.85% solution) 100.0mL Preparation of Dubos Oleic Albumin Complex: Add bovine serum albumin and oleic acid to 100.0mL of NaCl solution. Mix thoroughly. Filter sterilize. Penicillin Solution: Composition per 10.0mL: Penicillin 10,000U Preparation of Penicillin Solution: Add penicillin to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except Dubos oleic albumin complex and penicillin solution, to distilled/deionized water and bring volume to 970.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°– 50°C. Aseptically add sterile Dubos oleic albumin complex and penicillin solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Allow tubes to cool in a slanted position. Use: For the isolation of Mycobacterium tuberculosis and determining its sensitivity to chemotherapeutic agents. Dubos Oleic HiVeg Agar Base with Serum and Glycerol Composition per liter: Agar 15.0g Na 2 HPO 4 2.5g L-Asparagine 1.0g KH 2 PO 4 1.0g Plant hydrolysate 0.5g Ferric ammonium citrate 0.05g MgSO 4 0.01g CaCl 2 0.5mg CuSO 4 0.1mg ZnSO 4 0.1mg Glycerol 50.0mL Bovine serum or bovine albumin V 20.0mL pH 6.5 ± 0.2 at 25°C Source: This medium, without bovine serum or glycerol, is available as a premixed powder from HiMedia. Preparation of Medium: Add components, except bovine serum, 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 45°–50°C. Aseptically add 20.0mL sterile bovine serum or bovine serum albumin. Mix thoroughly. Aseptically distribute into sterile tubes. Use: For the cultivation of Mycobacterium tuberculosis and other Mycobacterium species. Dubos Oleic HiVeg Broth Base with Antibiotic and Oleic Albumin Supplement Composition per liter: Na 2 HPO 4 2.5g Asparagine 1.0g KH 2 PO 4 1.0g Plant hydrolysate 0.5g Ferric ammonium citrate 0.05g MgSO 4 0.01g CaCl 2 0.5mg CuSO 4 0.1mg ZnSO 4 0.1mg Penicllin solution 10.0mL pH 6.5 ± 0.2 at 25°C Source: This medium, without penicillin and oleic albumin supplement, is available as a premixed powder from HiMedia. Penicillin Solution: Composition per 10.0mL: Penicillin 10,000U Preparation of Penicllin Solution: Add penicillin to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Oleic Albumin Supplement: Composition per 100.0mL: Oleic acid 10.0g Albumin fraction IV 4.0g NaCl 0.68g NaOH 8.0mg Preparation of Oleic Albumin Supplement: Add components to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except penicllin solution and oleic albumin supplement, to distilled/deionized water and bring volume to 880.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°– 50°C. Aseptically add 10.0mL sterile penicillin solution and 100.0mL sterile oleic albumin solution. Mix thoroughly. Aseptically distribute into sterile tubes. Use: For the cultivation of Mycobacterium tuberculosis and other Mycobacterium species. Dubos Salts Agar Composition per liter: Agar 15.0g K 2 HPO 4 1.0g KCl 0.5g MgSO 4 ·7H 2 O 0.5g NaNO 3 0.5g FeSO 4 ·7H 2 O 0.01g Filter paper strips, sterile variable pH 7.2 ± 0.2 at 25°C Preparation of Medium: Add components, except filter paper strips, 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. Pour into sterile Petri dishes. Aseptically place sterile filter paper strips onto the surface of the solidified medium. Use: For the cultivation and maintenance of Alteromonas species, Cellvibrio mixtus, Cellvibrio species, Cytophaga aurantiaca, Cyto- phaga hutchinsonii, Pseudomonas species, and Sporocytophaga myxo- coccoides. Dubos Salts Agar with 1% Sodium Chloride Composition per liter: Agar 15.0g NaCl 10.0g © 2010 by Taylor and Francis Group, LLC 612 Dubos Salts Agar with Yeast Extract K 2 HPO 4 1.0g KCl 0.5g MgSO 4 ·7H 2 O 0.5g NaNO 3 0.5g FeSO 4 ·7H 2 O 10.0mg Filter paper strips 1 strip per tube 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. Autoclave for 15 min at 15 psi pressure–121°C. Allow tubes to cool in a slanted position. Aseptically add a strip of sterile filter paper to the surface of each slant. Use: For the cultivation of Cytophaga species. Dubos Salts Agar with Yeast Extract Composition per liter: Agar 15.0g K 2 HPO 4 1.0g Yeast extract 0.5g KCl 0.5g MgSO 4 ·7H 2 O 0.5g NaNO 3 0.5g FeSO 4 ·7H 2 O 0.01g Filter paper strips, sterile variable pH 7.2 ± 0.2 at 25°C Preparation of Medium: Add components, except filter paper strips, 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. Pour into sterile Petri dishes. Aseptically place sterile filter paper strips onto the surface of the solidified medium. Use: For the cultivation and maintenance of Cellulomonas species and Cellvibrio species. Dubos Salts Broth Composition per liter: K 2 HPO 4 1.0g KCl 0.5g MgSO 4 ·7H 2 O 0.5g NaNO 3 0.5g FeSO 4 ·7H 2 O 0.01g Filter paper strips, sterile variable pH 7.2 ± 0.2 at 25°C Preparation of Medium: Add components, except filter paper strips, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes containing a filter paper strip (filter paper strip should protrude above the surface of the broth). Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of Cytophaga aurantiaca and Pseudomonas species. Dubos Salts Broth with Yeast Extract Composition per liter: K 2 HPO 4 1.0g Yeast extract 0.5g KCl 0.5g MgSO 4 ·7H 2 O 0.5g NaNO 3 0.5g FeSO 4 ·7H 2 O 0.01g 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 Cytophaga aurantiaca. Dubos Salts Broth with Yeast Extract Composition per liter: K 2 HPO 4 1.0g Yeast extract 0.5g KCl 0.5g MgSO 4 ·7H 2 O 0.5g NaNO 3 0.5g FeSO 4 ·7H 2 O 0.01g Filter paper strips, sterile variable pH 7.2 ± 0.2 at 25°C Preparation of Medium: Add components, except filter paper strips, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes containing a filter paper strip (filter paper strip should protrude above the surface of the broth). Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of Cellulomonas species and Cellvibrio species. Dubos Salts Medium (DSMZ Medium 1161) Composition per liter: Agar 15.0g K 2 HPO 4 1.0g MgSO 4 ·7H 2 O 0.5g KCl 0.5g NaNO 3 0.5g FeSO 4 ·7H 2 O 10.0mg Filter paper Variable pH 7.0 ± 0.2 at 25°C Preparation of Medium: Add components, except filter paper, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.0. Gently heat while stirring 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. For tubes allow to solidify on a slant. When the agar has solidified, place a strip of sterile filter paper on to the surface of each slope. Inoculate on to the filter paper. Use: For the cultivation of Cellvibrio spp. Ducreyi Medium, Revised See: Haemophilus ducreyi Medium, Revised Dulaney Slants Composition per liter: Egg yolks 50.0mL Locke solution 50.0mL Locke Solution: Composition per 100.0mL: NaCl 0.9g Glucose 0.25g KCl 0.042g © 2010 by Taylor and Francis Group, LLC Dunkelberg Maintenance Medium 613 CaCl 2 ·2H 2 O 0.024g Na 2 CO 3 0.02g Preparation of Locke Solution: Add components to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Aseptically remove the yolks from 5–8 day old hen egg embryos. Add an equal volume of sterile Locke solution containing sterile glass beads. Mix thoroughly to homogenize. Asepti- cally distribute into sterile tubes. Inspissate tubes in a slanted position at 80°C (moist heat) for 15 min. Use: For the cultivation of Calymmatobacter granulomatis from clin- ical specimens. Dulcitol Selenite Broth (Selenite-F Broth with Dulcitol) Composition per liter: NaH 2 PO 4 10.0g Peptic digest of animal tissue 5.0g Dulcitol 4.0g HNaO 3 Se 4.0g pH 7.0 ± 0.2 at 25°C Source: This medium is available from HiMedia. Caution: Sodium hydrogen selenite is a very toxic, corrosive agent and causes teratogenicity. Upon contact with skin, wash immediately with a lot of water. Preparation of Medium: Add sodium hydrogen selenite to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Add remaining components. Mix thoroughly. Gently heat if needed to get all compoents to dissolve. Distribute into tubes or flasks. Sterilize in a boiling water bath or free flowing steam for 10 min. Excessive heating is detrimental. Do not autoclave. Use: For the selective enrichment of Salmonella species. Duncan-Strong Sporulation Medium, Modified (DS Sporulation Medium, Modified) (Sporulation Medium, Modified) Composition per liter: Proteose peptone 15.0g Na 2 HPO 4 ·7H 2 O 10.0g Raffinose 4.0g Yeast extract 4.0g Sodium thioglycolate 1.0g pH 7.8 ± 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. Adjust pH to 7.8 with filter-sterilized 0.66M Na 2 CO 3 . Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and induction of sporulation of Clostridium perfringens. Dung Extract Agar (DSMZ Medium 781) Composition per liter: Agar 15.0g Malt extract 5.0g (NO 3 ) 2 ·4H 2 O 0.72g MgSO 4 ·7H 2 O 0.5g K 2 HPO 4 0.25g Peptone 0.1g Dung extract 100.0mL pH 6.9 ± 0.2 at 25°C Dung Extract: Composition per 150.0mL: Horse dung variable Preparation of Dung Extract: Add an average sized piece of horse dung to150.0mL water. Gently heat and bring to boiling. Boil for 2 hr in a water bath. Filter. Use immediately. 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 Panaeolus cyanescens. Dunkelberg Agar See: Peptone Starch Dextrose Agar Dunkelberg Carbohydrate Medium, Modified Composition per 100.0mL: Proteose peptone No. 3 1.5g Carbohydrate 1.0g Na 2 HPO 4 ·2H 2 O 0.207g Phenol Red 0.055g NaH 2 PO 4 ·H 2 O 0.038g Horse serum 5.0mL pH 7.4 ± 0.2 at 25°C Preparation of Medium: Add components, except horse serum, to distilled/deionized water and bring volume to 95.0mL. For carbohydrate, use glucose, maltose, or starch. Mix thoroughly. Filter sterilize. Asepti- cally add sterile horse serum. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation and differentiation of Gardnerella vaginalis based on its ability to ferment glucose, maltose, or starch. Dunkelberg Maintenance Medium Composition per liter: Proteose peptone No. 3 20.0g Soluble starch 10.0g Agar 8.0g Glucose 2.0g Na 2 HPO 4 1.0g NaH 2 PO 4 1.0g pH 6.8 ± 0.2 at 25°C Preparation of Medium: Add starch to approximately 100.0mL of cold distilled/deionized water. Mix thoroughly. Add starch solution to 400.0mL of boiling distilled/deionized water. Add remaining components. Mix thoroughly. Bring volume to 1.0L with distilled/deionized water. Distribute into screw-capped tubes. Autoclave for 12 min at 8 psi pressure–112°C. Use: For the cultivation and maintenance of Gardnerella vaginalis. © 2010 by Taylor and Francis Group, LLC 614 Dunkelberg Semisolid Carbohydrate Fermentation Medium Dunkelberg Semisolid Carbohydrate Fermentation Medium Composition per liter: Proteose peptone No. 3 20.0g Carbohydrate 10.0g Agar 5.0g Bromcresol Purple solution 1.0mL pH 7.4 ± 0.2 at 25°C Bromcresol Purple Solution: Composition per 10.0mL: Bromcresol Purple 0.16g Ethanol (95% solution) 10.0mL Preparation of Bromcresol Purple Solution: Add Bromcresol Purple to 10.0mL of ethanol. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. For carbohydrate, use glucose, maltose, or starch. Mix thoroughly. Gently heat and bring to boiling. Filter sterilize. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation and differentiation of Gardnerella vaginalis based on its ability to ferment glucose, maltose, or starch. DV Medium Composition per 100.1mL: NaCl 1.8g MgSO 4 ·7H 2 O 0.5g Tris-HCl 100.0mg KCl 60.0mg NaNO 3 50.0mg Na 2 SiO 3 ·9H 2 O 20.0mg CaCl 2 10.0mg Nitrilotriacetic acid (NTA) 10.0mg K 2 HPO 4 3.0mg FeCl 2 0.01mg Vitamin B 12 0.3 μg Metal solution 3.0mL Vitamin solution 0.1mL Seawater, charcoal filtered 100.0mL Metal Solution: Composition per 25.0mL: EDTA 25.0mg MnCl 2 1.0mg H 3 BO 3 0.5mg FeCl 2 0.25mg ZnCl 2 125.0μg CoCl 2 25.0μg Preparation of Metal Solution: Add components to distilled/deionized water and bring volume to 25.0mL. Mix thoroughly. Vitamin Solution: Composition per 100.0mL: Inositol 100.0mg Thymine 50.0mg Orotic acid 26.0mg Thiamine·HCl 20.0mg Calcium D-(+)-pantothenate 10.0mg Nicotinic acid 10.0mg Putrescine·2HCl 4.0mg Pyridoxine·2HCl 4.0mg Pyridoxamine·2HCl 2.0mg p-Aminobenzoic acid 1.0mg Riboflavin 0.5mg Folic acid 0.025mg Biotin 50.0μg Folinic acid 20.0μg Vitamin B 12 5.0μg Preparation of Vitamin Solution: Add components to distilled/ deionized water and bring volume to 25.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except vitamin solution, to charcoal-filtered seawater and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Aseptical- ly add 0.1mL of sterile vitamin solution. Mix thoroughly. Aseptically distribute into sterile, screw-capped tubes or flasks. Use: For the cultivation of Dunaliella tertiolecta and Trichosphaer- ium species. DYA with Calcium Carbonate Composition per liter: Glucose 50.0g Agar 20.0g CaCO 3 5.0g Yeast extract 5.0g Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation of Dekkera intermedia, Dekkera bruxellensis, Dekkera abstinens, Dekkera anomala, Dekkera custersiana, Dekkera lambica, and Dekkera naardenensis. DYAA See: Dextrose Yeast Asparagine Agar DYAA Cellulose Composition per liter: Agar 20.0g Cellulose 5.0g Glucose 5.0g K 2 HPO 4 1.0g L-Asparagine 0.5g MgSO 4 ·7H 2 O 0.5g (NH 4 ) 2 SO 4 0.5g Yeast extract 0.5g CaCl 2 0.1g Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation of Bipolaris sacchari and Drechslera biseptata. DYAA Cellulose Malt Composition per liter: Agar 20.0g Malt extract 10.0g Cellulose 5.0g Glucose 5.0g © 2010 by Taylor and Francis Group, LLC . sterile Petri dishes or leave in tubes. Use: For the maintenance of lactic acid bacteria and differentiation of bacteria on the basis of lactose fermentation. Bacteria that ferment lactose appear. sterile Petri dishes or leave in tubes. Use: For the maintenance of lactic acid bacteria and differentiation of bacteria on the basis of lactose fermentation. Bacteria that ferment lactose appear. sterile Petri dishes or leave in tubes. Use: For the maintenance of lactic acid bacteria and differentiation of bacteria on the basis of lactose fermentation. Bacteria that ferment lactose appear