Cytophaga agarovorans Agar 475 Agar 2.5g L-Cystine 0.5g Na 2 SO 3 0.5g Phenol Red 17.0mg Carbohydrate solution 50.0mL pH 7.3 ± 0.2 at 25°C Source: This medium, without carbohydrate solution, is available as a premixed powder from HiMedia. Carbohydrate Solution: Composition per 100.0mL: Carbohydrate 10.0g Preparation of Carbohydrate Solution: Add carbohydrate to distilled/deionized water and bring volume to 100.0mL. Adonitol, ara- binose, cellobiose, glucose, dulcitol, fructose, galactose, inositol, lac- tose, maltose, mannitol, raffinose, rhamnose, salicin, sorbitol, sucrose, trehalose, xylose, or other carbohydrates may be used. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except carbohydrate solution, 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. Cool to 50°C. Aseptically add 50.0mL of sterile carbohydrate solution. Mix thor- oughly. Pour into sterile Petri dishes or leave in tubes. Use: For fermentation studies of various bacteria. CYT Agar Composition per liter: Agar 15.0g Pancreatic digest of casein 1.0g CaCl 2 ·2H 2 O 0.5g MgSO 4 ·7H 2 O 0.5g Yeast extract 0.5g pH 7.2 ± 0.2 at 25°C Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the isolation and cultivation of Cytophaga species, Herpeto- siphon species, Saprospira species, and Flexithrix species. Cytophaga Agarase Agar (ATCC Medium 793) Composition per liter: Agar 15.0g KH 2 PO 4 1.0g MgSO 4 ·7H 2 O 0.5g NH 4 Cl 0.5g CaCl 2 ·H 2 O 0.02g Vishniac and Santer trace element mixture 0.2mL pH 7.2 ± 0.2 at 25°C Vishniac and Santer Trace Element Mixture: Composition per liter: Ethylenediamine tetraacetic acid (EDTA) 50.0g ZnSO 4 ·7H 2 O 22.0g CaCl 2 5.54g MnCl 2 ·4H 2 O 5.06g FeSO 4 ·7H 2 O 4.99g CoCl 2 ·6H 2 O 1.61g CuSO 4 ·5H 2 O 1.57g (NH 4 ) 6 Mo 7 O 24 ·4H 2 O 1.1g Preparation of Vishniac and Santer Trace Element Mix- ture: Add components to distilled/deionized water and bring volume to 1.0L. Adjust pH to 6.0 with KOH. Mix thoroughly. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Adjust pH to 7.2. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure– 121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Cytophaga flevensis. Cytophaga Agarase Broth Composition per liter: Agar 1.0g KH 2 PO 4 1.0g MgSO 4 ·7H 2 O 0.5g NH 4 Cl 0.5g CaCl 2 ·H2O 0.02g Vishniac and Santer trace element mixture 0.2mL pH 7.2 ± 0.2 at 25°C Vishniac and Santer Trace Element Mixture: Composition per liter: Ethylenediamine tetraacetic acid (EDTA) 50.0g ZnSO 4 ·7H 2 O 22.0g CaCl 2 5.54g MnCl 2 ·4H 2 O 5.06g FeSO 4 ·7H 2 O 4.99g CoCl 2 ·6H 2 O 1.61g CuSO 4 ·5H 2 O 1.57g (NH 4 ) 6 Mo 7 O 24 ·4H 2 O 1.1g Preparation of Vishniac and Santer Trace Element Mix- ture: Add components to distilled/deionized water and bring volume to 1.0L. Adjust pH to 6.0 with KOH. Mix thoroughly. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Adjust pH to 7.2. Mix thoroughly. Dis- tribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure– 121°C. Use: For the cultivation of Cytophaga flevensis. Cytophaga agarovorans Agar (LMG Medium 99) Composition per 1001.0mL: NaCl 30.0g Agar 15.0g KH 2 PO 4 1.0g MgSO 4 ·7H 2 O 1.0g NH 4 Cl 1.0g Yeast extract 1.0g CaCl 2 ·2H 2 O 50.0mg FeCl 3 ·H 2 O 1.25mg Glucose solution 10.0mL NaHCO 3 solution 10.0mL Na 2 S·9H 2 O solution 1.0mL Glucose Solution: Composition per 10.0mL: D-Glucose 1.0g © 2010 by Taylor and Francis Group, LLC 476 Cytophaga agarovorans Broth Preparation of Glucose Solution: Add glucose to distilled/deion- ized water and bring volume to 10.0mL. Mix thoroughly. Filter steril- ize. NaHCO 3 Solution: Composition per 100.0mL: NaHCO 3 5.0g Preparation of NaHCO 3 Solution: Add NaHCO 3 to distilled/de- ionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Na 2 S·9H 2 O Solution: Composition per 10.0mL: Na 2 S·9H 2 O 1.0g Preparation of Na 2 S·9H 2 O Solution: Add Na 2 S·9H 2 O to dis- tilled/deionized water and bring volume to 10.0mL. Filter sterilize. Preparation of Medium: Add components, except glucose solu- tion, NaHCO 3 solution, and Na 2 S·9H 2 O solution, to distilled/deionized water and bring volume to 980.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Aseptically add 10.0mL of sterile glucose solution, 10.0mL of sterile NaHCO 3 solution, and 1.0mL of sterile Na 2 S·9H 2 O solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation of Cytophaga agarovorans. Cytophaga agarovorans Broth (LMG Medium 99) Composition per 1001.0mL: NaCl 30.0g KH 2 PO 4 1.0g MgSO 4 ·7H 2 O 1.0g NH 4 Cl 1.0g Yeast extract 1.0g Agar 1.0g CaCl 2 ·2H 2 O 50.0mg FeCl 3 ·H 2 O 1.25mg Glucose solution 10.0mL NaHCO 3 solution 10.0mL Na 2 S·9H 2 O solution 1.0mL Glucose Solution: Composition per 10.0mL: D-Glucose 1.0g Preparation of Glucose Solution: Add glucose to distilled/deion- ized water and bring volume to 10.0mL. Mix thoroughly. Filter steril- ize. NaHCO 3 Solution: Composition per 100.0mL: NaHCO 3 5.0g Preparation of NaHCO 3 Solution: Add NaHCO 3 to distilled/de- ionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Na 2 S·9H 2 O Solution: Composition per 10.0mL: Na 2 S·9H 2 O 1.0g Preparation of Na 2 S·9H 2 O Solution: Add Na 2 S·9H 2 O to dis- tilled/deionized water and bring volume to 10.0mL. Filter sterilize. Preparation of Medium: Add components, except glucose solu- tion, NaHCO 3 solution, and Na 2 S·9H 2 O solution, to distilled/deionized water and bring volume to 980.0mL. Add 1.0g of agar as a detoxifying agent. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically add 10.0mL of sterile glucose solution, 10.0mL of sterile NaHCO 3 solution, and 1.0mL of sterile Na 2 S·9H 2 O solution. Mix thor- oughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation and maintenance of Cytophaga agarovorans. Cytophaga fermentans Medium Composition per liter: NaCl 30.0g Agar 5.0g NaHCO 3 5.0g KH 2 PO 4 1.0g NH 4 Cl 1.0g MgCl 2 ·6H 2 O 0.5g Yeast extract 0.3g Na 2 S·9H 2 O 0.1g CaCl 2 0.04g Ferric citrate (4mM solution) 5.0mL Trace elements solution 2.0mL pH 7.0 ± 0.2 at 25°C Trace Elements Solution: Composition per 100.0mL: H 3 BO 3 0.28g MnSO 4 ·6H 2 O 0.21g Na 2 MoO 4 ·2H 2 O 0.075g Zn(NO 3 ) 2 ·6H 2 O 0.025g CoCl 2 ·6H 2 O 0.02g Cu(NO 3 ) 2 ·3H 2 O 0.02g Preparation of Trace Elements Solution: Add components to distilled/deionized water and bring volume to 100.0mL. Mix thorough- ly. 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. Use: For the cultivation of agar-digesting Cytophaga fermentans. Cytophaga hutchinsonii Agar Composition per liter: Agar 15.0g Pancreatic digest of casein 3.0g CaCl 2 ·2H 2 O 1.36g Yeast extract 1.0g Cellobiose solution 50.0mL pH 7.2 ± 0.2 at 25°C Cellobiose Solution: Composition per 50.0mL: D-Cellobiose 6.0g Preparation of Cellobiose Solution: Add cellobiose to distilled/ deionized water and bring volume to 50.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except cellobiose solu- tion, 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 50°–55°C. Aseptically add 50.0mL of © 2010 by Taylor and Francis Group, LLC Cytophaga Medium 477 sterile cellobiose solution. Mix thoroughly. Pour into sterile Petri dish- es or distribute into sterile tubes. Use: For the cultivation and maintenance of Cytophaga aurantiaca and Cytophaga hutchinsonii. Cytophaga hutchinsonii Broth Composition per liter: Pancreatic digest of casein 3.0g CaCl 2 ·2H 2 O 1.36g Yeast extract 1.0g Filter paper strips 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 thor- oughly. Gently heat and bring to boiling. Distribute in 5.0mL volumes into tubes. Add a strip of filter paper about 7.0cm in length to each tube. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of Cytophaga aurantiaca and Cytophaga hutchinsonii. Cytophaga Marine Medium (DSMZ Medium 172) Composition per liter: NaCl 24.7g Agar 15.0g MgSO 4 ·7H 2 O 6.3g MgCl 2 ·6H 2 O 4.6g CaCl 2 ·2H 2 O 1.2g Yeast extract 1.0g Tryptone 1.0g KCl 0.7g Sodium bicarbonate solution 10.0mL Calcium chloride solution 10.0mL pH 7.0 ± 0.2 at 25°C Sodium Bicarbonate Solution: Composition per 10.0mL: NaHCO 3 0.2g Preparation of Sodium Bicarbonate Solution: Add NaHCO 3 to distilled/deionized water and bring volume to 10.0mL. Mix thorough- ly. Autoclave for 15 min at 15 psi pressure–121°C. Calcium Chloride Solution: Composition per 10.0mL: CaCl 2 ·2H 2 O 1.2g Preparation of Calcium Chloride Solution: Add CaCl 2 ·2H 2 O to distilled/deionized water and bring volume to 10.0mL. Mix thorough- ly. Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: Add components, except sodium bicar- bonate and calcium chloride solution, to distilled/deionized water and bring volume to 900.0mL. 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 of sterile bicarbonate solution and 10.0mL sterile calcium chloride solution. Adjust pH to 7.2. Mix thor- oughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of Cellulophaga lytica, Cy- tophaga latercula, Marinilabilia salmonicolor, Saprospira grandis, Cytophaga marinoflava, Persicobacter diffluens, Flammeovirga apri- ca, Flexibacter tractuosus, Microscilla spp., Marinilabilia salmonicol- or, Flexibacter litoralis, Flexithrix dorotheae, and Cellulophaga lytica. Cytophaga Medium Composition per liter: NaCl 30.0g Agar 15.0g KH 2 PO 4 1.0g NH 4 Cl 1.0g Yeast extract 1.0g MgSO 4 0.5g CaCl 2 0.04g FeCl 3 ·6H 2 O 1.25mg NaHCO 3 solution 100.0mL Glucose solution 10.0mL Na 2 S·9H 2 O solution 1.0mL Glucose Solution: Composition per 100.0mL: Glucose 10.0g Preparation of Glucose Solution: Add glucose to distilled/deion- ized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. NaHCO 3 Solution: Composition per 100.0mL: NaHCO 3 5.0g Preparation of NaHCO 3 Solution: Add NaHCO 3 to distilled/de- ionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Na 2 S·9H 2 O Solution: Composition per 100.0mL: Na 2 S·9H 2 O 10.0g Preparation of Na 2 S·9H 2 O Solution: Add Na 2 S·9H 2 O to dis- tilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: Add components, except NaHCO 3 solu- tion, glucose solution, and Na 2 S·9H 2 O solution, to distilled/deionized water and bring volume to 889.0mL. Autoclave for 15 min at 15 psi pres- sure–121°C. Cool to 50°C. Aseptically add sterile NaHCO 3 solution, sterile glucose solution, and sterile Na 2 S·9H 2 O solution. Mix thorough- ly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation of Cytophaga agarovorans. Cytophaga Medium (ATCC Medium 420) Composition per liter: NaCl 20.0g Yeast extract 10.0g Agar 3.0g MgSO 4 ·7H 2 O 1.0g NH 4 Cl 1.0g K 2 HPO 4 0.2g FeCl 3 1.0μg pH 7.5 ± 0.2 at 25°C Preparation of Medium: Add components to tap water and bring volume to 1.0L. Adjust pH to 7.5. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of Cytophaga fermentans. © 2010 by Taylor and Francis Group, LLC 478 Cytophaga Medium Cytophaga Medium (ATCC Medium 1299) Composition per liter: Agar 11.0g Pancreatic digest of casein 0.5g Yeast extract 0.5g Beef extract 0.2g Sodium acetate 0.2g pH 7.2 ± 0.2 at 25°C Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Cytophaga species and Flavobacterium branchiophilum. Cytophaga Medium, Modified Composition per liter: NaCl 20.0g Agar 15.0g Yeast extract 10.0g MgSO 4 ·7H 2 O 1.0g NH 4 Cl 1.0g K 2 HPO 4 0.2g FeCl 3 1.0μg pH 7.5 ± 0.2 at 25°C Preparation of Medium: Add components to tap water and bring volume to 1.0L. Adjust pH to 7.5. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of Cytophaga fermentans. Cytophaga Spirochete Medium See: Cytophaga Medium See: Spirochete Medium Cytosine Nutrient Agar Composition per liter: Agar 15.0g Pancreatic digest of gelatin 5.0g Beef extract 3.0g Cytosine 20.0mg 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 of Escherichia coli. Cystine Tellurite Blood Agar Composition per 120.0mL: Heart infusion agar 100.0mL K 2 TeO 3 solution 15.0mL Sheep blood 5.0mL L-Cystine 5.0mg pH 7.4 ± 0.2 at 25°C Heart Infusion Agar: Composition per liter: Beef heart, infusion from 500.0g Agar 20.0g Tryptose 10.0g Yeast extract 5.0g NaCl 5.0g Preparation of Heart Infusion Agar: Add components to dis- tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Au- toclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. K 2 TeO 3 Solution: Composition per 100.0mL: K 2 TeO 3 0.3g Preparation of K 2 TeO 3 Solution: Add K 2 TeO 3 to distilled/deion- ized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Caution: Potassium tellurite is toxic. Preparation of Medium: Add sterile K 2 TeO 3 solution, sterile, de- fibrinated sheep blood, and sterile, solid L-cystine to sterile, cooled heart infusion agar. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the isolation, differentiation, and cultivation of Corynebacte- rium diphtheriae. Corynebacterium diphtheriae appears as dark gray to black colonies. CYU 2% Composition per liter: Basal solution 960.0mL Yeast extract solution 30.0mL Trace metal solution 10.0mL Basal Solution: Composition per 960.0mL: Glucose 20.0g K 2 HPO 4 9.2g Urea 5.0g (NH 4 ) 2 S0 4 2.2g KH 2 PO 4 1.3g Trisodium citrate·2H 2 O 1.0g MgSO 4 ·7H 2 O solution 1.66mL MgSO 4 ·7H 2 O Solution: Composition per 10.0mL: MgSO 4 ·7H 2 O 2.5g Preparation of MgSO 4 ·7H 2 O Solution: Add MgSO 4 ·7H 2 O to distilled/deionized water and bring volume to 10.0mL. Mix thorough- ly. Preparation of Basal Solution: Add components to distilled/de- ionized water and bring volume to 960.0mL. Mix thoroughly. Filter sterilize. Yeast Extract Solution: Composition per 100.0mL: Yeast extract 10.0g Preparation of Yeast Extract Solution: Add yeast extract to dis- tilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. © 2010 by Taylor and Francis Group, LLC Czapek Agar with 20% Sucrose 479 Trace Metal Solution: Composition per liter: CaCl 2 ·2H 2 O 3972.0mg FeSO 4 ·7H 2 O 1250.0mg H 3 BO 3 1140.0mg ZnSO 4 ·7H 2 O 882.0mg CuCl 2 ·5H 2 O 157.0mg MnCl 2 ·4H 2 O 140.0mg NaMoO 4 ·2H 2 O 119.0mg Vanadyl sulfate·2H 2 O 100.0mg CsCl 2 ·6H 2 O 49.0mg FeCl 3 ·6H 2 O 29.0mg CdSO 4 ·8H 2 O 10.0mg Ni(NO 3 ) 2 ·6H 2 O 10.0mg HCl, concentrated 10.0mL Preparation of Trace Metal Solution: Add 10.0mL of concen- trated HCl to 900.0mL of distilled/deionized water. Mix thoroughly. Add remaining components in the order shown. Mix thoroughly after adding each component. Bring volume to 990.0mL with distilled/de- ionized water. Mix thoroughly. Filter sterilize. Preparation of Medium: Aseptically combine 960.0mL of sterile basal solution, 30.0mL of sterile yeast extract solution, and 30.0mL of sterile trace metal solution. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation of Neospongicoccum excentricum. CZA See: Czapek Agar CZA200 See: Czapek Agar with 20% Sucrose Czapek Agar (ATCC Medium 312) Composition per liter: Sucrose 30.0g Agar 15.0g NaNO 3 3.0g K 2 HPO 4 1.0g KCl 0.5g MgSO 4 ·7H 2 O 0.5g FeSO 4 ·7H 2 O 0.01g pH 7.3 ± 0.2 at 25°C Preparation of Medium: Add components, except sucrose, to dis- tilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Distribute into tubes or flasks. In a separate flask, add sucrose to dis- tilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave both solutions separately for 15 min at 15 psi pressure– 121°C. Cool to 50°C. Combine the sterile solutions. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of Streptomyces species. For the cultivation of Actinoplanaceae. Czapek Agar See: Czapek Yeast Autolysate Agar Czapek Agar with Peptone (ATCC Medium 522) Composition per liter: Sucrose 30.0g Agar 15.0g Peptone 5.0g NaNO 3 3.0g K 2 HPO 4 1.0g KCl 0.5g MgSO 4 ·7H 2 O 0.5g FeSO 4 ·7H 2 O 0.01g pH 7.3 ± 0.2 at 25°C Preparation of Medium: Add components, except sucrose, to dis- tilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Distribute into tubes or flasks. In a separate flask, add sucrose to dis- tilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave both solutions separately for 15 min at 15 psi pressure– 121°C. Cool to 50°C. Combine the sterile solutions. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of Streptomyces species. For the cultivation of Actinoplanaceae. Czapek Agar with Sucrose Composition per liter: Sucrose 170.0g Agar 15.0g NaNO 3 3.0g K 2 HPO 4 1.0g KCl 0.5g MgSO 4 ·7H 2 O 0.5g FeSO 4 ·7H 2 O 0.01g pH 7.3 ± 0.2 at 25°C Preparation of Medium: Add components, except sucrose, to dis- tilled/deionized water and bring volume to 700.0mL. Mix thoroughly. In a separate flask, add sucrose to distilled/deionized water and bring volume to 300.0mL. Mix thoroughly. Autoclave both solutions sepa- rately for 15 min at 15 psi pressure–121°C. Cool to 45–50°C. Combine the sterile solutions. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation of Aspergillus echinulatus, Aspergillus flavi- pes, Aspergillus penicilloides, numerous Eurotium species, Penicil- lium citreonigrum, and Penicillium thomii. Czapek Agar with 20% Sucrose (CZA200) Composition per liter: Sucrose 200.0g Agar 15.0g NaNO 3 3.0g K 2 HPO 4 1.0g KCl 0.5g MgSO 4 ·7H 2 0 0.5g FeSO 4 ·7H 2 O 0.01g pH 7.3 ± 0.2 at 25°C Preparation of Medium: Add components, except sucrose, to dis- tilled/deionized water and bring volume to 700.0mL. Mix thoroughly. In a separate flask, add sucrose to distilled/deionized water and bring volume to 300.0mL. Mix thoroughly. Autoclave both solutions sepa- rately for 15 min at 15 psi pressure–121°C. Cool to 45–50°C. Combine © 2010 by Taylor and Francis Group, LLC 480 Czapek Dox Agar the sterile solutions. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation of Aspergillus repens (Eurotium repens). Czapek Dox Agar Composition per liter: Sucrose 30.0g Agar 15.0g NaNO 3 3.0g K 2 HPO 4 1.0g MgSO 4 ·7H 2 O 0.5g KCl 0.5g FeSO 4 ·7H 2 O 0.01g pH 7.3 ± 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. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Actinoplanes species, Amorphosporangium auranticolor, Ampullariella species, Spiril- lospora albida, and Streptomyces armeniacus. Czapek Dox Agar Composition per liter: Sucrose 30.0g Agar 15.0g NaNO 3 2.0g K 2 HPO 4 1.0g MgSO 4 ·7H 2 O 0.5g KCl 0.5g FeSO 4 ·7H 2 O 0.01g 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. 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 Actinoplanes species, Amorphosporangium auranticolor, Ampullariella species, Spiril- lospora albida, and Streptomyces armeniacus. Czapek Dox Agar with 3% Glucose Composition per liter: Glucose 30.0g Agar 15.0g NaNO 3 3.0g K 2 HPO 4 1.0g KCl 0.5g MgSO 4 ·7H 2 O 0.5g FeSO 4 ·7H 2 O 0.01g pH 7.3 ± 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 Microbispora rosea and Streptomyces species. Czapek Dox Agar with 20% Sucrose Composition per liter: Sucrose 200.0g Agar 20.0g NaNO 3 2.0g K 2 HPO 4 1.0g KCl 0.5g MgSO 4 ·7H 2 O 0.5g FeSO 4 0.01g ZnSO 4 0.01g CuSO 4 0.005g 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 20 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Aspergillus brunneus, Aspergillus equitis, Aspergillus hollandicus, Aspergillus nidulellus, Aspergillus reptans, and Aspergillus rubrobrunneus. Czapek Dox Agar, Modified Composition per liter: Sucrose 30.0g Agar 12.0g NaNO 3 2.0g Magnesium glycerophosphate 0.5g KCl 0.5g K 2 SO 4 0.35g FeSO 4 0.01g pH 6.8 ± 0.2 at 25°C Source: This medium is available as a premixed powder from Oxoid Unipath and HiMedia. 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. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of numerous fungal species. For chlamydospore production by Candida albicans. Czapek Dox Broth Composition per liter: Sucrose 30.0g NaNO 3 3.0g K 2 HPO 4 1.0g MgSO 4 ·7H 2 O 0.5g KCl 0.5g FeSO 4 ·7H 2 O 0.01g pH 7.3 ± 0.2 at 25°C Source: This medium is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation and maintenance of a variety of fungal and bacterial species that can use nitrate as sole nitrogen source. © 2010 by Taylor and Francis Group, LLC Czapek Solution Agar with Sucrose 481 Czapek Dox Liquid Medium, Modified Composition per liter: Sucrose 30.0g NaNO 3 2.0g Magnesium glycerophosphate 0.5g KCl 0.5g K 2 SO 4 0.35g FeSO 4 0.01g pH 6.8 ± 0.2 at 25°C Source: This medium is available as a premixed powder from Oxoid Unipath. 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 fungi and bacteria capable of utilizing sodium nitrate as the sole source of nitrogen. Czapek Malt Agar Composition per liter: Malt extract 40.0g Sucrose 30.0g Agar 15.0g KNO 3 2.0g K 2 HPO 4 1.0g KCl 0.5g MgSO 4 ·7H 2 O 0.5g FeSO 4 ·7H2O 0.1g pH 6.8 ± 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. Pour into sterile Petri dishes or leave in tubes. Use: For the isolation and cultivation of saprophytic fungi. Czapek Peptone Agar Composition per liter: Sucrose 30.0g Agar 15.0g Peptone 5.0g KNO 3 2.0g Yeast extract 2.0g K 2 HPO 4 1.0g KCl 0.5g MgSO 4 ·7H 2 O 0.5g FeSO 4 ·7H 2 O 0.01g pH 7.3 ± 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 of Actinoplanes campanulatus, Actinoplanes dig- itatis, Actinoplanes italicus, Actinoplanes lobatus, Actinoplanes regularis, Actinoplanes utahensis, Ampullariella campanulata, Micromonospora brunnea, Micromonospora purpurea, Micromonospora purpureochro- mogenes, Micromonospora species, Microtetraspora glauca, Micromono- spora chalcea, Nocardia brevicatena, Pilimelia anulata, Pilimelia terevasa, Planomonospora parontospora, Promicromonospora citrea, Saccharomonospora caesia, Saccharomonospora viridis, Spirillospora albida, Sporichthya polymorpha, Streptomyces yerevanensis, Thermoac- tinomyces thalpophilus, Thermoactinomyces vulgaris, and Thermomono- spora chromogena. Czapek Peptone Yeast Agar Composition per liter: Sucrose 30.0g Agar 15.0g Peptone 5.0g NaNO 3 3.0g Yeast extract 2.0g K 2 HPO 4 1.0g KCl 0.5g MgSO 4 ·7H 2 0 0.5g FeSO 4 ·7H 2 O 0.01g pH 7.3 ± 0.2 at 25°C Preparation of Medium: Add components, except sucrose, to dis- tilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Distribute into tubes or flasks. In a separate flask, add sucrose to dis- tilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave both solutions separately for 15 min at 15 psi pressure– 121°C. Cool to 50°C. Combine the sterile solutions. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation of various actinomycetes. Czapek Solution Agar Composition per liter: Sucrose 30.0g Agar 15.0g NaNO 3 2.0g K 2 HPO 4 1.0g KCl 0.5g MgSO 4 ·7H 2 0 0.5g FeSO 4 ·7H 2 O 0.01g pH 7.3 ± 0.2 at 25°C Source: This medium is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. 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 Aspergillus, Penicillium, and other fungi. For the cultivation and maintenance of microorganisms that can utilize nitrate as sole nitrogen source. Czapek Solution Agar with Sucrose Composition per liter: Sucrose 200.0g Agar 20.0g NaNO 3 3.0g K 2 HPO 4 1.0g KCl 0.5g MgSO 4 ·7H 2 O 0.5g FeSO 4 ·7H 2 O 10.0mg 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. © 2010 by Taylor and Francis Group, LLC 482 Czapek Yeast Autolysate Agar Use: For the cultivation and maintenance of osmophilic fungi. Czapek Yeast Autolysate Agar (CYA Agar) (Czapek Agar) Composition per liter: Agar 15.0g Yeast extract 5.0g NaNO 3 3.0g K 2 HPO 4 1.0g KCl 0.5g MgSO 4 ·7H 2 O 0.5g FeSO 4 ·7H 2 O 0.01g Sucrose solution 100.0mL pH 7.3 ± 0.2 at 25°C Sucrose Solution: Composition per 100.0mL: Sucrose 30.0g Preparation of Sucrose Solution: Add sucrose to distilled/deion- ized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Preparation of Medium: Add components, except sucrose solution, to distilled/deionized water and bring volume to 900.0mL. Mix thor- oughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Aseptically add sterile sucrose solution. Mix thoroughly. Pour into sterile Petri dishes or leave in tubes. Use: For the isolation and cultivation of heat-resistant filamentous fungi (molds) from foods. Czapek Yeast Extract Agar Composition per liter: Sucrose 30.0g Agar 15.0g Yeast extract 5.0g K 2 HPO 4 1.0g Czapek concentrate 10.0mL Czapek Concentrate: Composition per liter: NaNO 3 30.0 g KCl 5.0 g MgSO 4 ·7H 2 O 5.0 g FeSO 4 ·7H2O 0.1 g ZnSO 4 ·7H 2 O 0.1 g CuSO 4 ·5H 2 O 0.05 g Preparation of Czapek Concentrate: Add components to dis- tilled/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. 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 Aspergillus niger. Czapek Yeast Extract Agar Composition per liter: Sucrose 30.0g Agar 15.0g Yeast extract 5.0g NaNO 3 3.0g K 2 HPO 4 1.0g MgSO 4 ·7H 2 O 0.5g KCl 0.5g FeSO 4 ·7H 2 O 0.01g Trace metal solution 1.0mL pH 7.3 ± 0.2 at 25°C Trace Metal Solution: Composition per 100.0mL: ZnSO 4 ·7H 2 O 1.0g CuSO 4 ·5H 2 O 0.5g Preparation of Trace Metal Solution: Add components to 100.0mL distilled/deionized water. Mix thoroughly. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 6.2. Gently heat and bring to boiling. Distribute into tubes or flasks. Auto- clave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of a variety of fungal and bacterial species that can use nitrate as sole nitrogen source. Czapek Yeast Extract Agar Composition per liter: Sucrose 30.0g Agar 15.0g Yeast extract 5.0g K 2 HPO 4 1.0g KNO 3 0.3g KCl 0.05g MgSO 4 ·7H 2 O 0.05g FeSO 4 ·7H2O 1.0mg ZnSO 4 ·7H 2 O 1.0mg CuSO 4 ·5H 2 O 0.5mg pH 6.8 ± 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. Pour into sterile Petri dishes or leave in tubes. Use: For the isolation and cultivation of Aspergillus niger. CZYA See: Czapek Yeast Autolysate Agar DA Medium Composition per liter: NaCl 116.9g Agar 15.0g Tris-HCl 6.024g NaHCO 3 1.68g MgSO 4 ·7H 2 O 1.232g KNO 3 0.505g CaCl 2 0.033g KH 2 PO 4 0.014g H 3 BO 3 6.0mg MnCl 2 ·4H 2 O 99.0μg ZnCl 2 14.0μg CoCl 2 ·6H 2 O 4.76μg © 2010 by Taylor and Francis Group, LLC Davis and Mingioli Medium, Modified 483 CuCl 2 ·2H 2 O 34.0ng FeCl 3 solution 50.0mL pH 7.5 ± 0.2 at 25°C FeCl 3 Solution: Composition per 50.0mL: EDTA 5.84mg FeCl 3 0.32mg Preparation of FeCl 3 Solution: Add components to distilled/de- ionized water and bring volume to 50.0mL. Mix thoroughly. Preparation of Medium: Add components to distilled/deionized wa- ter and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into screw-capped tubes. Autoclave for 15 min at 15 psi pressure–121°C. Allow to cool in a slanted position. Use: For the cultivation of Dunaliella bardawil. Dap Nutrient Agar Composition per liter: Urea 20.0g Agar 15.0g Peptone 5.0g Meat extract 3.0g DL-α,ε-Diaminopimelic acid 0.1g 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. Adjust pH to 7.0. Autoclave 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 Bacillus megaterium. Dap Nutrient Agar Composition per liter: Urea 20.0g Agar 15.0g Peptone 5.0g Meat extract 3.0g DL-α,ε-Diaminopimelic acid 0.1g MnSO 4 ·H 2 O 10.0mg 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. Adjust pH to 7.0. Autoclave for 15 min at 15 psi pressure– 121°C. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the sporulation of Bacillus megaterium. Davis and Mingioli Glucose Minimal Medium Composition per liter: Agar 15.0g K 2 HPO 4 7.0g KH 2 PO 4 3.0g (NH 4 ) 2 SO 4 1.0g Sodium citrate·3H 2 O 0.5g MgSO 4 ·7H 2 O 0.1g L-Arginine 0.02g L-Tryptophan 0.02g Glucose solution 10.0mL pH 7.0 ± 0.2 at 25°C Glucose Solution: Composition per 10.0mL: Glucose 2.0g Preparation of Glucose Solution: Add glucose to distilled/deion- ized water and bring volume to 10.0mL. Mix thoroughly. Filter steril- ize. Preparation of Medium: Add components, except glucose solu- tion, to distilled/deionized water and bring volume to 990.0mL. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure– 121°C. Cool to 45°–50°C. Aseptically add sterile glucose solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation and maintenance of Escherichia coli. Davis and Mingioli Medium A Composition per liter: K 2 HPO 4 7.0g KH 2 PO 4 3.0g (NH 4 ) 2 SO 4 1.0g Sodium citrate·3H 2 O 0.5g MgSO 4 ·7H 2 O 0.1g Glucose solution 10.0mL Amino acid solution 10.0mL pH 7.0 ± 0.2 at 25°C Glucose Solution: Composition per 10.0mL: Glucose 2.5g Preparation of Glucose Solution: Add glucose to distilled/deion- ized water and bring volume to 10.0mL. Mix thoroughly. Filter steril- ize. Amino Acid Solution: Composition per 10.0mL: L-Leucine 40.0mg L-Histidine 20.0mg L-Methionine 20.0mg Preparation of Amino Acid Solution: Add components to dis- tilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except glucose solu- tion and amino acid solution, to distilled/deionized water and bring volume to 980.0mL. 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 of sterile glucose solution and 10.0mL of sterile amino acid so- lution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation of Escherichia coli. Davis and Mingioli Medium, Modified Composition per liter: K 2 HPO 4 7.0g KH 2 PO 4 2.0g (NH 4 ) 2 SO 4 1.0g Na citrate·2H 2 O 0.5g MgSO 4 ·7H 2 O 0.1g Glucose solution 10.0mL © 2010 by Taylor and Francis Group, LLC 484 Davis and Mingioli Medium, Modified Streptomycin solution 10.0mL Additives solution 10.0mL pH 7.2 ± 0.2 at 25°C Glucose Solution: Composition per 10.0mL: Glucose 4.0g Preparation of Glucose Solution: Add glucose to distilled/deion- ized water and bring volume to 10.0mL. Mix thoroughly. Filter steril- ize. Streptomycin Solution: Composition per 10.0mL: Streptomycin 4.0g Preparation of Streptomycin Solution: Add streptomycin to dis- tilled/deionized water and bring volume to 10.0mL. Mix well. Filter sterilize. Additives Solution: Composition per 10.0mL: DL-Threonine 0.1g DL- or L-Leucine 0.1g Thiamine·HCl 0.5mg Preparation of Additives Solution: Add components to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components—except glucose solu- tion, streptomycin solution, and additives solution—to distilled/deion- ized water and bring volume to 970.0mL. Mix thoroughly. Adjust pH to 7.2. Gently heat and bring to boiling. Autoclave for 25 min at 15 psi pres- sure–121°C. Cool to 45°–50°C. Aseptically add sterile glucose solution, streptomycin solution, and additives solution. Mix thoroughly. Asepti- cally distribute into sterile tubes or flasks. Use: For the cultivation and maintenance of Escherichia coli. Davis and Mingioli Medium, Modified Composition per liter: Agar 15.0g K 2 HPO 4 7.0g Lactose 2.0g (NH 4 ) 2 SO 4 1.0g KH 2 PO 4 0.91g MgSO 4 ·7H 2 O 0.1g Tap water 10.0mL 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. Adjust pH to 7.0. 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 Escherichia coli. Davis and Mingioli Medium with Proline Composition per liter: K 2 HPO 4 7.0g KH 2 PO 4 3.0g Glucose 2.0g (NH 4 ) 2 SO 4 1.0g L-Proline 0.5g Sodium citrate·3H 2 O 0.5g MgSO 4 ·7H 2 O 0.1g Glucose solution 1.0mL pH 7.0 ± 0.2 at 25°C Glucose Solution: Composition per 100.0mL: Glucose 20.0g Preparation of Glucose Solution: Add glucose to distilled/deion- ized water and bring volume to 100.0mL. Mix thoroughly. Filter ster- ilize. Preparation of Medium: Add components, except glucose solu- tion, to distilled/deionized water and bring volume to 999.0mL. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure– 121°C. Cool to 25°C. Aseptically add 1.0mL of sterile glucose solu- tion. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation and maintenance of Escherichia coli. Davis and Mingioli Medium with Vitamin B 1 and Asparagine Composition per liter: K 2 HPO 4 7.0g KH 2 PO 4 3.0g (NH 4 ) 2 SO 4 1.0g Sodium citrate·3H 2 O 0.5g L-Asparagine 0.4g MgSO 4 ·7H 2 O 0.1g Vitamin B 1 0.1mg Glucose solution 1.0mL pH 7.0 ± 0.2 at 25°C Glucose Solution: Composition per 100.0mL: Glucose 20.0g Preparation of Glucose Solution: Add glucose to distilled/deion- ized water and bring volume to 100.0mL. Mix thoroughly. Filter ster- ilize. Preparation of Medium: Add components, except glucose solu- tion, to distilled/deionized water and bring volume to 999.0mL. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure– 121°C. Cool to 25°C. Aseptically add 1.0mL of sterile glucose solu- tion. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation and maintenance of Escherichia coli. Davis Supplemented Minimal Medium Composition per liter: Agar 15.0g K 2 HPO 4 7.0g KH 2 PO 4 3.0g Casein hydrolysate 2.0g Yeast extract 2.0g (NH 4 ) 2 SO 4 1.0g Sodium citrate·3H 2 O 0.5g MgSO 4 ·7H 2 O 0.1g Glucose solution 20.0mL pH 7.0 ± 0.2 at 25°C Glucose Solution Composition per 100.0mL: Glucose 10.0g © 2010 by Taylor and Francis Group, LLC . 100.0mg CsCl 2 ·6H 2 O 49. 0mg FeCl 3 ·6H 2 O 29.0mg CdSO 4 ·8H 2 O 10.0mg Ni(NO 3 ) 2 ·6H 2 O 10.0mg HCl, concentrated 10.0mL Preparation of Trace Metal Solution: Add 10.0mL of concen- trated HCl to 900.0mL of distilled/deionized. thoroughly. Filter sterilize. Preparation of Medium: Aseptically combine 960.0mL of sterile basal solution, 30.0mL of sterile yeast extract solution, and 30.0mL of sterile trace metal solution. Aseptically. pressure–121°C. Cool to 50°–55°C. Aseptically add 10.0mL of sterile glucose solution, 10.0mL of sterile NaHCO 3 solution, and 1.0mL of sterile Na 2 S·9H 2 O solution. Mix thoroughly. Pour into