Charcoal Agar 345 Solution 1: Composition per liter: MgCl 2 ·6H 2 O 40.0g KCl 4.0g CaCl 2 ·2H 2 O 0.4g pH 7.4 ± 0.2 at 25°C Preparation of Solution 1: 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. Adjust pH to 7.4. Dis- tribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure– 121°C. Use: For the cultivation of Haloarcula vallismortis. Chaetomium Medium Composition per liter: Agar 15.0g NaNO 3 2.0g K 2 HPO 4 1.2g MgSO 4 ·7H 2 O 0.5g KCl 0.5g KH 2 PO 4 0.14g Yeast extract 0.02g Fe 2 (SO 4 ) 3 ·H 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. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Allow agar to cool. Place a strip of sterile filter paper on top of agar and inocu- late filter strip. Use: For the growth and maintenance of Chaetomium species. Chalquist’s Antigen Medium, Modified Composition per liter: Soluble starch 0.5g Pancreatic digest of casein 0.05g L-Cysteine·HCl·H 2 O 0.01g NAD (nicotinamide adeninedinucleotide) 0.01g PPLO broth without Crystal Violet 90.0mL Swine serum, inactivated 10.0mL Phenol Red (1% solution) 0.25mL pH 7.6 ± 0.2 at 25°C PPLO Broth without Crystal Violet: Composition per 500.0mL: Beef heart, infusion from 11.52g Peptone 2.32g NaCl 1.15g Source: PPLO broth without Crystal Violet is available as a premixed powder from BD Diagnostic Systems. Preparation of PPLO Broth without Crystal Violet: Add components to distilled/deionized water and bring volume to 500.0mL. 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 Mycoplasma synoviae. Chapman Stone Agar Composition per liter: (NH 4 ) 2 SO 4 75.0g NaCl 55.0g Gelatin 30.0g Agar 15.0g D-Mannitol 10.0g Pancreatic digest of casein 10.0g K 2 HPO 4 5.0g Yeast extract 2.0g pH 7.0 ± 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. Autoclave for 10 min at 15 psi pressure–121°C. Pour into sterile Petri dishes while the medi- um is still hot. Add 25.0mL of medium per Petri dish. Use: For the isolation of staphylococci from a variety of specimens. Charcoal Agar Composition per liter: Beef heart, solids from infusion 500.0g Agar 18.0g Peptone 10.0g Soluble starch 10.0g NaCl 5.0g Charcoal, activated, acid washed 4.0g Yeast extract 3.5g 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. Gently heat and bring to boiling with frequent stirring. Autoclave for 15 min at 15 psi pres- sure–121°C. Cool to 45°–50°C. Pour into sterile Petri dishes or leave in tubes. Shake flask while dispensing to keep charcoal in suspension. Allow tubes to cool in a slanted position. Use: For the cultivation and maintenance of fastidious microorgan- isms, especially Bordetella pertussis, for vaccine production. Charcoal Agar Composition per liter: Agar 12.0g Beef extract 10.0g Peptone 10.0g Starch 10.0g NaCl 5.0g Charcoal 4.0g Nicotinic acid 0.001g pH 7.4 ± 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. Gently heat and bring to boiling with frequent stirring. Autoclave for 15 min at 15 psi pres- sure–121°C. Cool to 45°–50°C. This medium may be enriched by the addition of blood. Pour into sterile Petri dishes or distribute into tubes. Shake flask while dispensing to keep charcoal in suspension. © 2010 by Taylor and Francis Group, LLC 346 Charcoal Agar Base, HiVeg with Blood and Selective Supplement Use: For the cultivation and isolation of various bacteria; with the addition of blood, for the cultivation of fastidious bacteria. Charcoal Agar Base, HiVeg with Blood and Selective Supplement Composition per liter: Agar 18.0g Plant infusion 12.0g Plant peptone 10.0g Starch 10.0g NaCl 5.0g Charcoal 4.0g Yeast extract 3.5g Sheep blood, defibrinated 50.0mL Selective supplement solution 10.0mL pH 7.3 ± 0.2 at 25°C Source: This medium, without blood or selective supplement, is available as a premixed powder from HiMedia. Selective Supplement Solution: Composition per 10.0mL: Cefazolin 40.0mg Preparation of Selective Supplement Solution: Add cefazolin to distilled/deionized water and bring volume to 10.0mL. Mix thor- oughly. Filter sterilize. Preparation of Medium: Add components, except blood and se- lective supplement solution, to distilled/deionized water and bring vol- ume to 940.0mL. Mix thoroughly. Gently heat and bring to boiling with frequent stirring. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add 50.0 mL sterile sheep blood and 10.0mL sterile selective supplement solution. Mix thoroughly. Pour into sterile Petri plates or distribute into sterile tubes. Use: For the cultivation and isolation of various bacteria; with the addition of blood, for the cultivation of fastidious bacteria. For the cul- tivation of Bordetella pertussis for vaccine production and the mainte- nance of stock cultures. Charcoal Agar with Horse Blood Composition per liter: Agar 12.0g Beef extract 10.0g Peptone 10.0g Starch 10.0g NaCl 5.0g Charcoal, bacteriological 4.0g Nicotinic acid 1.0mg Horse blood, defibrinated 100.0mL pH 7.4 ± 0.2 at 25°C Preparation of Medium: Add components to distilled/deionized water and bring volume to 900.0L. Mix thoroughly. Gently heat and bring to boiling with frequent stirring. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 80°C. Aseptically add 100.0mL of sterile, defi- brinated horse blood. Maintain at 80°C for 10 min to form chocolate agar. Pour into sterile Petri dishes or distribute into tubes. Shake flask while dispensing to keep charcoal in suspension. Use: For the cultivation and isolation of Haemophilus influenzae. Charcoal Agar with Horse Blood and Cepahalexin Composition per liter: Agar 12.0g Beef extract 10.0g Peptone 10.0g Starch 10.0g NaCl 5.0g Charcoal 4.0g Nicotinic acid 1.0mg Horse blood, defibrinated 100.0mL Cephalexin solution 10.0mL pH 7.4 ± 0.2 at 25°C Cephalexin Solution: Composition per 10.0mL: Cephalexin 0.04g Preparation of Cephalexin Solution: Add cephalexin to dis- tilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except cephalexin so- lution and horse blood, to distilled/deionized water and bring volume to 890.0L. Mix thoroughly. Gently heat and bring to boiling with fre- quent stirring. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add 100.0mL of sterile, defibrinated horse blood and 10.0mL of sterile cephalexin solution. Pour into sterile Petri dishes or distribute into tubes. Shake flask while dispensing to keep charcoal in suspension. Use: For the cultivation and isolation of Bordetella pertussis. Charcoal Agar Slants See: Diphasic Medium for Amoeba Charcoal Blood Agar Base, HiVeg with Blood Penicillin and Damidodiphenyl Hydrochloride Composition per liter: Agar 12.0g Plant extract 10.0g Plant peptone 10.0g Starch 10.0g NaCl 5.0g Charcoal 4.0g Yeast extract 3.5g Sheep blood, defibrinated 50.0mL Penicillin solution 3.0mL Diamidodiphenylamine hydrochloride solution 3.0mL pH 7.5 ± 0.2 at 25°C Source: This medium, without blood or selective supplement, is available as a premixed powder from HiMedia. Penicllin Solution: Composition per 10.0mL: Penicillin 1000 units Preparation of Penicillin Solution: Add penicillin to distilled/de- ionized water and bring volume to 10.0mL. Mix thoroughly. Filter ster- ilize. Diamidodiphenylamine Hydrochloride Solution: Composition per 10.0mL: Diamido hydrochloride 0.01g Diphenylamine hydrochloride 0.01g © 2010 by Taylor and Francis Group, LLC CHCA Salts Medium 347 Preparation of Diamidodiphenylamine Hydrochloride So- lution: Add components to distilled/deionized water and bring vol- ume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except blood, penicil- lin solution, and diamidodiphenylamine hydrochloride solution, to dis- tilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Gently heat and bring to boiling with frequent stirring. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add 50.0 mL sterile sheep blood and 3.0 mL sterile penicillin solution and 3.0 mL sterile diamidodiphenylamine hydrochloride solution. Mix thoroughly. Pour into sterile Petri plates or distribute into sterile tubes. Use: For the cultivation and isolation of various bacteria; with the addition of blood, for the cultivation of fastidious bacteria. For the cul- tivation of Bordetella pertussis for vaccine production and the mainte- nance of stock cultures. Charcoal Blood Medium Composition per liter: Beef heart, solids from infusion 500.0g Agar 18.0g Peptone 10.0g Soluble starch 10.0g NaCl 5.0g Charcoal, activated, acid washed 4.0g Yeast extract 3.5g Horse or sheep blood, defibrinated 100.0mL Cephalexin solution 10.0mL pH 7.4 ± 0.2 at 25°C Cephalexin Solution: Composition per 10.0mL: Cephalexin 0.04g Preparation of Cephalexin Solution: Add cephalexin to dis- tilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except blood and ce- phalexin solution, to distilled/deionized water and bring volume to 890.0mL. 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 blood and cephalexin solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the cultivation of Haemophilus influenzae. Charcoal HiVeg Agar Base with Niacin, Blood, and Selective Supplement Composition per liter: Agar 12.0g Plant extract 10.0g Plant peptone No. 2 10.0g Starch 10.0g NaCl 5.0g Charcoal 4.0g Niacin 1.0mg Sheep blood, defibrinated 50.0mL Selective supplement solution 10.0mL pH 7.4 ± 0.2 at 25°C Source: This medium, without blood or selective supplement, is available as a premixed powder from HiMedia. Selective Supplement Solution: Composition per 10.0mL: Cefazolin 40.0mg Preparation of Selective Supplement Solution: Add cefazolin to distilled/deionized water and bring volume to 10.0mL. Mix thor- oughly. Filter sterilize. Preparation of Medium: Add components, except blood and se- lective supplement solution, to distilled/deionized water and bring vol- ume to 940.0mL. Mix thoroughly. Gently heat and bring to boiling with frequent stirring. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add 50.0 mL sterile sheep blood and 10.0mL sterile selective supplement solution. Mix thoroughly. Pour into sterile Petri plates or distribute into sterile tubes. Use: For the cultivation and isolation of Bordetella pertussis and Hae- mophilus influenzae. For the cultivation of Bordetella pertussis for vaccine production and the maintenance of stock cultures. Charcoal Yeast Extract Agar See: CYE Agar Charcoal Yeast Extract Agar, Buffered See: CYE Agar, Buffered Charcoal Yeast Extract Diphasic Blood Culture Medium See: Legionella pneumophila Medium Chase’s Medium SP Composition per liter: Agar 10.0g Proteose peptone 10.0g K 2 HPO 4 2.0g (NH 4 ) 2 SO 4 1.0g Sucrose solution 100.0mL pH 6.5 ± 0.2 at 25°C Sucrose Solution: Composition per 100.0mL: Sucrose 10.0g Preparation of Sucrose Solution: Add sucrose to distilled/deion- ized water and bring volume to 100.0mL. Mix thoroughly. Filter ster- ilize. Preparation of Medium: Add components, except sucrose solu- tion, to tap water and bring volume to 900.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pres- sure–121°C. Cool to 45°–50°C. Aseptically add 100.0mL of sterile sucrose solution. Mix thoroughly. Pour into sterile Petri dishes or dis- tribute into sterile tubes. Use: For the cultivation and maintenance of ATCC strain 13949. CHCA Salts Medium (Cyclohexane Carboxylic Acid Salts Medium) Composition per liter: K 2 HPO 4 3.5g KH 2 PO 4 1.5g Cyclohexane carboxylic acid 1.0g NH 4 NO 3 1.0g MgSO 4 ·7H 2 O 0.5g FeSO 4 ·7H 2 O 0.1g Yeast extract 0.1g © 2010 by Taylor and Francis Group, LLC 348 Cheese Agar CaCl 2 ·2H 2 O 0.01g Na 2 MoO 2 ·2H 2 O 0.01g ZnSO 4 ·7H 2 O 0.01g 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. 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 bacteria that can utilize cyclohexane carboxylic acid as a carbon source. For the cultivation and maintenance of Arthrobacter globiformis. Cheese Agar Composition per liter: Cheese, ripened 100.0g NaCl 50.0g Agar 15.0g Peptone 10.0g Potassium citrate 10.0g Sodium oxalate 2.0g pH 7.4 ± 0.2 at 25°C Preparation of Medium: Add the cheese and potassium citrate to distilled/deionized water and bring volume to 300.0mL. Gently heat and bring to 50°C to separate the fat. Discard the fat. In a separate flask, add the remaining components to distilled/deionized water and bring volume to 700.0mL. Gently heat and bring to boiling. Add the 300.0mL of aqueous suspension of cheese solids. Adjust pH to 7.4. Au- toclave for 25 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or distribute into sterile tubes. Use: For the isolation and cultivation of Brevibacterium linens. Cherry Agar, CBS Formula Composition per liter: Pulp of sour stone cherries 200.0g Agar 20.0g pH 3.8 ± 0.2 at 25°C Preparation of Medium: Add 1.0L of distilled/deionized water to pulp of sour stone cherries. Gently heat and bring to boiling. Simmer gently for 2 hr. Strain through cloth. Autoclave filtrate for 30 min at 6 psi pressure–110°C. Add agar to distilled/deionized water and bring volume to 800.0mL. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. To sterile agar solution, add 200.0mL of sterile cherry filtrate. Distribute into sterile tubes. Autoclave for 5 min at 15 psi pressure–121°C. Use: For the cultivation of various fungi. Cherry Decoction Agar Composition per liter: Agar 20.0g Sucrose 10.0g Cherry extract 90.0mL pH 3.8–4.6 at 25°C Preparation of Medium: Add agar to distilled/deionized water and bring volume to 500.0mL. Gently heat and bring to boiling. In a sepa- rate flask, add sucrose and cherry extract to distilled/deionized water and bring volume to 500.0mL. Mix thoroughly. Combine the two solu- tions. Adjust pH to 3.8–4.6. Distribute into tubes or flasks. Sterilize by tyndallization at 100°C for 30 min on 3 consecutive days. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation and maintenance of Acremonium sclerotigenum, Amylostereum laevigatum, Calcarisporium arbuscula, Helicoma mor- ganii, Helicoon richonis, Inonotus radiatus, Phellinus igniarius, Phoma leveillei, Taphrina californica, and Taphrina populina. CHI 1776 Medium Composition per liter: Pancreatic digest of casein 25.0g Yeast extract 7.5g Glucose solution 25.0mL Tris·HCl buffer, 1M, pH 7.5 20.0mL Diaminopimelic acid solution 10.0mL Thymidine solution 10.0mL MgCl 2 solution 5.0mL Glucose Solution: Composition per 100.0mL: D-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. Diaminopimelic Acid Solution: Composition per 100.0mL: meso-Diaminopimelic acid 1.0g Preparation of Diaminopimelic Acid Solution: Add meso-di- aminopimelic acid to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Thymidine Solution: Composition per 100.0mL: Thymidine 0.4g Preparation of Thymidine Solution: Add thymidine to distilled/ deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. MgCl 2 Solution: Composition per 100.0mL: MgCl 2 9.52g Preparation of MgCl 2 Solution: Add MgCl 2 to distilled/deion- ized 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 glucose solu- tion, diaminopimelic acid solution, thymidine solution, and MgCl 2 so- lution, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Aseptical- ly add 25.0mL of sterile glucose solution, 10.0mL of sterile diamin- opimelic acid solution, 10.0mL of sterile thymidine solution, and 5.0mL of sterile MgCl 2 solution, Mix thoroughly. Distribute into sterile tubes or flasks. Use: For the cultivation of Escherichia coli. Chicken Soup Broth Composition per 5.0L: Chicken 2.5kg Peppercorns 6 Cloves 3 Bay leaf 2 Celery, stalks including leaves 2 © 2010 by Taylor and Francis Group, LLC Chitin Agar 349 Onion, large 1 Carrot 1 Dill, fresh 1/4 cup NaCl 0.1g Preparation of Medium: Add a nice, whole chicken to a large pot. Add enough tap water to cover the chicken by about 1 in. Stud the whole, peeled onion with the three cloves. Add the onion and remain- ing ingredients to the pot. Rapidly heat and bring to boiling. Lower heat to a simmer and cook for 1 to 1.5 hr. Remove the chicken and vegeta- bles from the broth. Remove skin and bones from the chicken. Cut up the meat into 1-inch pieces. Return the meat to the broth. If desired, slice the carrot and celery and return them to the broth. Use: For the growth and nutrition of microbiologists. China Blue Lactose Agar Composition per liter: Agar 12.0g Lactose 10.0g Peptone 5.0g NaCl 5.0g Beef extract 3.0g China Blue 0.375g pH 7.0 ± 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. 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, differentiation, and enumeration of bacteria from dairy products. Lactose-fermenting bacteria appear as blue colo- nies. Nonlactose-fermenting bacteria appear as colorless colonies. China Blue Lactose Agar Composition per liter: Agar 15.0g Lactose 10.0g Peptic digest of animal tissue 5.0g NaCl 5.0g Plant extract 3.0g China Blue 0.3g pH 7.0 ± 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 cultivation, differentiation, and enumeration of bacteria from dairy products. Lactose-fermenting bacteria appear as blue colo- nies. Nonlactose-fermenting bacteria appear as colorless colonies. China Blue Lactose HiVeg Agar Composition per liter: Agar 15.0g Lactose 10.0g Plant peptone 5.0g NaCl 5.0g Plant extract 3.0g China Blue 0.3g pH 7.0 ± 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 cultivation, differentiation, and enumeration of bacteria from dairy products. Lactose-fermenting bacteria appear as blue colo- nies. Nonlactose-fermenting bacteria appear as colorless colonies. Chitin Agar Composition per liter: Agar 15.0g Chitin, precipitated 3.0g (NH 4 ) 2 SO 4 2.0g Na 2 HPO 4 1.1g KH 2 PO 4 0.7g MgSO 4 ·7H 2 O 0.2g FeSO 4 1.0mg MnSO 4 1.0mg Chitin, Precipitated: Composition per 2.5L: Chitin 40.0g HCl, concentrated 400.0mL Preparation of Chitin, Precipitated: Add chitin to 400.0mL of cold concentrated HCl. Add this solution to 2.0L of distilled/deionized water at 5°C. Filter the solution through Whatman #1 filter paper. Dia- lyze the precipitated chitin against tap water for 12 hr. Adjust the pH to 7.0 with KOH. 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. Chitin Agar Composition per liter: Agar 20.0g Chitin 4.0g K 2 HPO 4 0.7g MgSO 4 ·7H 2 O 0.5g KH 2 PO 4 0.3g FeSO 4 ·7H 2 O 0.01g MnCl 2 ·4H 2 O 0.001g ZnSO 4 ·7H 2 O 0.001g pH 8.0 ± 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. Use: For the selective isolation and cultivation of streptomycetes. © 2010 by Taylor and Francis Group, LLC 350 Chlamydia Growth Medium Chlamydia Growth Medium Composition per 500.0mL: Eagle minimum essential medium with Earle salts, 10X 50.0mL Fetal calf serum 50.0mL L-Glutamine solution 5.0mL pH 7.4 ± 0.2 at 25°C Eagle Minimum Essential Medium with Earle Salts, 10X: Composition per liter: NaCl 6.8g Glucose 1.0g KCl 0.4g CaCl 2 ·2H 2 O 0.2g MgCl 2 ·6H 2 O 0.2g NaH 2 PO 4 0.15g L-Arginine 0.1g L-Lysine 0.06g L-Isoleucine 0.05g L-Leucine 0.05g L-Threonine 0.05g L-Valine 0.05g L-Tyrosine 0.04g L-Phenylalanine 0.03g L-Histidine 0.03g L-Cystine 0.02g L-Methionine 0.02g L-Tryptophan 0.01g i-Inositol 2.0mg Calcium pantothenate 1.0mg Choline chloride 1.0mg Folic acid 1.0mg Nicotinamide 1.0mg Pyridoxal 1.0mg Thiamine·HCl 1.0mg Riboflavin 0.1mg Preparation of Eagle Minimum Essential Medium with Earle Salts, 10X: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.4 with 7.5% Na 2 CO 3 solution. Filter sterilize. Glutamine Solution: Composition per 100.0mL: L-Glutamine 2.92g NaCl (0.85% solution) 100.0mL Preparation of Glutamine Solution: Add the glutamine to the 0.85% NaCl solution. Mix thoroughly. Filter sterilize. Preparation of Medium: Aseptically combine 50.0mL of sterile Ea- gle minimum essential medium with Earle salts, 10X, 50.0mL of fetal calf serum, and 5.0mL of sterile glutamine solution. Bring volume to 500.0mL with sterile distilled/deionized water. Mix thoroughly. Aseptically distrib- ute into sterile tubes or flasks. Use: For the cultivation of Chlamydia species. Chlamydia Isolation Medium Composition per 500.0mL: Eagle minimum essential medium with Earle salts, 10X 50.0mL Fetal calf serum 50.0mL Selective supplement 10.0mL L-Glutamine solution 5.0mL pH 7.4 ± 0.2 at 25°C Eagle Minimum Essential Medium with Earle Salts, 10X: Composition per liter: NaCl 6.8g Glucose 1.0g KCl 0.4g CaCl 2 ·2H 2 O 0.2g MgCl 2 ·6H 2 O 0.2g NaH 2 PO 4 0.15g L-Arginine 0.1g L-Lysine 0.06g L-Isoleucine 0.05g L-Leucine 0.05g L-Threonine 0.05g L-Valine 0.05g L-Tyrosine 0.04g L-Phenylalanine 0.03g L-Histidine 0.03g L-Cystine 0.02g L-Methionine 0.02g L-Tryptophan 0.01g i-Inositol 2.0mg Calcium pantothenate 1.0mg Choline chloride 1.0mg Folic acid 1.0mg Nicotinamide 1.0mg Pyridoxal 1.0mg Thiamine·HCl 1.0mg Riboflavin 0.1mg Preparation of Eagle Minimum Essential Medium with Earle Salts, 10X: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.4 with 7.5% Na 2 CO 3 solution. Filter sterilize. Selective Supplement: Composition per 10.0mL: Glucose 0.594g Vancomycin 0.05g Gentamicin 0.01g Amphotericin B 2.0mg Cycloheximide 2.0mg Preparation of Selective Supplement: Add components to dis- tilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Caution: Cycloheximide is toxic. Avoid skin contact or aerosol for- mation and inhalation. Glutamine Solution: Composition per 100.0mL: L-Glutamine 2.92g NaCl (0.85% solution) 100.0mL Preparation of Glutamine Solution: Add the glutamine to the 0.85% NaCl solution. Mix thoroughly. Filter sterilize. Preparation of Medium: Aseptically combine 50.0mL of sterile Eagle minimum essential medium with Earle salts, 10X, 50.0mL of fe- tal calf serum, 10.0mL of selective supplement, and 5.0mL of sterile glutamine solution. Bring volume to 500.0mL with sterile distilled/de- © 2010 by Taylor and Francis Group, LLC Chloramphenicol Ampicillin LB Medium 351 ionized water. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the isolation and cultivation of Chlamydia species. Chlamydomonas Enriched Medium Composition per liter: Agar 15.0g Sodium acetate, anhydrous 2.0g Tryptose 2.0g Yeast extract 2.0g 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 20 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Use: For the cultivation of Chlamydomonas reinhardtii. Chlamydomonas Mutant Agar Composition per liter: Agar 18.0g K 2 HPO 4 1.162g Sodium acetate 1.0g NaH 2 PO 4 0.92g Sodium citrate 0.5g MgSO 4 ·7H 2 O 0.3g NH 4 NO 3 0.3g CaCl 2 0.04g FeCl 3 ·6H 2 O 0.01g Trace salts solution 10.0mL pH 6.8 ± 0.2 at 25°C Trace Salts Solution: Composition per liter: H 3 BO 3 0.15g LiSO 4 ·4H 2 O 0.1g MnSO 4 ·4H 2 O 0.04g CoCl 2 0.02g (NH 4 ) 6 Mo 7 O 24 0.015g CuSO 4 ·5H 2 O 0.004g Preparation of Trace Salts Solution: 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. Adjust pH to 6.8. 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 Chlamydomonas reinhardtii. Chlamydomonas Mutant Broth Composition per liter: K 2 HPO 4 1.162g Sodium acetate 1.0g NaH 2 PO 4 0.92g Sodium citrate 0.5g NH 4 NO 3 0.3g MgSO 4 ·7H 2 O 0.3g CaCl 2 0.04g FeCl 3 ·6H 2 O 0.01g Trace salts solution 10.0mL pH 6.8 ± 0.2 at 25°C Trace Salts Solution: Composition per liter: H 3 BO 3 0.15g LiSO 4 ·4H 2 O 0.1g MnSO 4 ·4H 2 O 0.04g CoCl 2 0.02g (NH 4 ) 6 Mo 7 O 24 0.015g CuSO 4 ·5H 2 O 0.004g Preparation of Trace Salts Solution: 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. Adjust pH to 6.8. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure– 121°C. Use: For the cultivation of Chlamydomonas reinhardtii. Chlamydospore Agar Composition per liter: Purified polysaccharide 20.0g Agar 15.0g KH 2 PO 4 1.0g (NH 4 ) 2 SO 4 1.0g Trypan Blue 0.1g Biotin 5.0μg pH 5.1 ± 0.2 at 25°C Source: This medium is available as a premixed powder from BD Di- agnostic Systems. Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat 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 differentiating Candida albicans from other Candida species on the basis of chlamydospore formation. Chloramphenicol Ampicillin LB Medium Composition per liter: NaCl 10.0g Pancreatic digest of casein 10.0g Yeast extract 5.0g Ampicillin solution 10.0mL Chloramphenicol solution 10.0mL pH 7.0 ± 0.2 at 25°C Ampicillin Solution: Composition per 10.0mL: Ampicillin 40.0mg Preparation of Ampicillin Solution: Add ampicillin to distilled/ deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize. Chloramphenicol Solution: Composition per 10.0mL: Chloramphenicol 5.0mg Preparation of Chloramphenicol Solution: Add chlorampheni- col to distilled/deionized water and bring volume to 10.0mL. Mix thor- oughly. Filter sterilize. Preparation of Medium: Add components, except ampicillin solu- tion and chloramphenicol solution, to distilled/deionized water and bring volume to 980.0mL. Mix thoroughly. Adjust pH to 7.0. Auto- © 2010 by Taylor and Francis Group, LLC 352 Chloramphenicol Erythromycin LB Medium clave for 15 min at 15 psi pressure–121°C. Aseptically add 10.0mL of sterile ampicillin solution and 10.0mL of sterile chloramphenicol solu- tion. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation and maintenance of Escherichia coli. Chloramphenicol Erythromycin LB Medium Composition per liter: NaCl 10.0g Pancreatic digest of casein 10.0g Yeast extract 5.0g Erythromycin 10.0mL Chloramphenicol 10.0mL pH 7.0 ± 0.2 at 25°C Erythromycin Solution: Composition per 10.0mL: Erythromycin 10mg Preparation of Erythromycin Solution: Add erythromycin to distilled/deionized water and bring volume to 10.0mL. Mix thorough- ly. Filter sterilize. Chloramphenicol Solution: Composition per 10.0mL: Chloramphenicol 5.0mg Preparation of Chloramphenicol Solution: Add chlorampheni- col to distilled/deionized water and bring volume to 10.0mL. Mix thor- oughly. Filter sterilize. Preparation of Medium: Add components, except erythromycin so- lution and chloramphenicol solution, to distilled/deionized water and bring volume to 980.0mL. Mix thoroughly. Adjust pH to 7.0. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically add 10.0mL of sterile ampicillin solution and 10.0mL of sterile chloramphenicol solution. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation and maintenance of Escherichia coli. Chloramphenicol L Broth Medium No. 1 Composition per liter: Pancreatic digest of casein 10.0g NaCl 5.0g Yeast extract 5.0g Glucose 1.0g Chloramphenicol solution 10.0mL pH 7.0 ± 0.2 at 25°C Chloramphenicol Solution: Composition per 10.0mL: Chloramphenicol 5.0mg Preparation of Chloramphenicol Solution: Add chlorampheni- col to distilled/deionized water and bring volume to 10.0mL. Mix thor- oughly. Filter sterilize. Preparation of Medium: Add components, except chlorampheni- col solution, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Adjust pH to 7.0. Autoclave for 15 min at 15 psi pres- sure–121°C. Aseptically add 10.0mL of sterile chloramphenicol solu- tion and 10.0mL of sterile chloramphenicol solution. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation and maintenance of Escherichia coli. Chloramphenicol L Broth Medium No. 2 Composition per liter: Pancreatic digest of casein 10.0g NaCl 5.0g Yeast extract 5.0g Glucose 1.0g Chloramphenicol solution 10.0mL pH 7.0 ± 0.2 at 25°C Chloramphenicol Solution: Composition per 10.0mL: Chloramphenicol 12.5mg Preparation of Chloramphenicol Solution: Add chlorampheni- col to distilled/deionized water and bring volume to 10.0mL. Mix thor- oughly. Filter sterilize. Preparation of Medium: Add components, except chlorampheni- col solution, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Adjust pH to 7.0. Autoclave for 15 min at 15 psi pres- sure–121°C. Aseptically add 10.0mL of sterile chloramphenicol solu- tion and 10.0mL of sterile chloramphenicol solution. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation and maintenance of Escherichia coli. Chloramphenicol L Broth Medium No. 3 Composition per liter: Pancreatic digest of casein 10.0g NaCl 5.0g Yeast extract 5.0g Glucose 1.0g Chloramphenicol solution 10.0mL pH 7.0 ± 0.2 at 25°C Chloramphenicol Solution: Composition per 10.0mL: Chloramphenicol 50.0mg Preparation of Chloramphenicol Solution: Add chlorampheni- col to distilled/deionized water and bring volume to 10.0mL. Mix thor- oughly. Filter sterilize. Preparation of Medium: Add components, except chlorampheni- col solution, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Adjust pH to 7.0. Autoclave for 15 min at 15 psi pres- sure–121°C. Aseptically add 10.0mL of sterile chloramphen icol solu- tion and 10.0mL of sterile chloramphenicol solution. Mix thoroughly. Aseptically distribute into sterile tubes or flasks. Use: For the cultivation and maintenance of Escherichia coli. Chloramphenicol LB Medium No.1 Composition per liter: NaCl 10.0g Pancreatic digest of casein 10.0g Yeast extract 5.0g Chloramphenicol 50.0μg Preparation of Medium: Adjust pH to 7.0. Autoclave at 15 psi pres- sure–121°C for 15 min. For solid medium, add 15.0g of agar. Use: For the cultivation and maintenance of Escherichia coli. Chloramphenicol LB Medium No. 2 Composition per liter: NaCl 10.0g Pancreatic digest of casein 10.0g © 2010 by Taylor and Francis Group, LLC Chlorella Broth without Glucose and Citrate 353 Yeast extract 5.0g Chloramphenicol solution 10.0mL pH 7.0 ± 0.2 at 25°C Chloramphenicol Solution: Composition per 10.0mL: Chloramphenicol 5.0mg Preparation of Chloramphenicol Solution: Add chlorampheni- col to distilled/deionized water and bring volume to 10.0mL. Mix thor- oughly. Filter sterilize. Preparation of Medium: Add components, except chlorampheni- col solution, to distilled/deionized water and bring volume to 990.0mL. Mix thoroughly. Adjust pH to 7.0. Autoclave for 15 min at 15 psi pres- sure–121°C. Aseptically add 10.0mL of sterile ampicillin solution and 10.0mL of sterile chloramphenicol solution. Mix thoroughly. Asepti- cally distribute into sterile tubes or flasks. Use: For the cultivation and maintenance of Escherichia coli. Chloramphenicol Yeast Glucose Agar Composition per liter: Glucose 20.0g Agar 14.9g Yeast extract 5.0g Chloramphenicol 0.1g pH 6.6 ± 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 selective enumeration of yeasts and molds in milk and milk products. It is also recommended by ISO Committee under the specifications ISO 7954:1987. Chlorella Agar Composition per liter: Agar 17.0g KNO 3 2.5g KH 2 PO 4 2.45g MgSO 4 ·7H 2 O 2.4g K 2 SO 4 0.217g FeSO 4 ·7H 2 O 1.5mg MnSO 4 ·H 2 O 1.4mg H 3 BO 3 0.28mg ZnSO 4 ·7H 2 O 0.22mg Na 2 MoO 4 ·2H 2 O 0.05mg CuSO 4 ·5H 2 O 0.0078mg Supplement solution 100.0mL pH 4.5 ± 0.2 at 25°C Source: This medium is available from HiMedia. Supplement Solution: Composition per 100.0mL: Glucose 10.0g Potassium citrate 32.0mg Preparation of Supplement Solution: Add components to dis- tilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except supplement so- lution, to distilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Aseptically add supplement solution. Mix thoroughly. Pour into Petri dishes or aseptically distribute into sterile tubes. Use: For the cultivation of Chlorella spp. Chlorella Broth Composition per liter: KNO 3 2.5g KH 2 PO 4 2.45g MgSO 4 ·7H 2 O 2.4g K 2 SO 4 0.217g FeSO 4 ·7H 2 O 1.5mg MnSO 4 ·H 2 O 1.4mg H 3 BO 3 0.28mg ZnSO 4 ·7H 2 O 0.22mg Na 2 MoO 4 ·2H 2 O 0.05mg CuSO 4 ·5H 2 O 0.0078mg Supplement solution 100.0mL pH 4.5 ± 0.2 at 25°C Source: This medium is available from HiMedia. Supplement Solution: Composition per 100.0mL: Glucose 10.0g Potassium citrate 32.0mg Preparation of Supplement Solution: Add components to dis- tilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: Add components, except supplement so- lution, to distilled/deionized water and bring volume to 900.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°C. Aseptically add supplement solution. Mix thoroughly. Asepti- cally distribute into sterile tubes. Use: For the cultivation of Chlorella spp. Chlorella Broth without Glucose and Citrate Composition per liter: KNO 3 2.5g KH 2 PO 4 2.45g MgSO 4 ·7H 2 O 2.4g K 2 SO 4 0.217g FeSO 4 ·7H 2 O 1.5mg MnSO 4 ·H 2 O 1.4mg H 3 BO 3 0.28mg ZnSO 4 ·7H 2 O 0.22mg Na 2 MoO 4 ·2H 2 O 0.05mg CuSO 4 ·5H 2 O 0.0078mg Supplement solution 100.0mL pH 4.5 ± 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. Use: For the cultivation of Chlorella spp. © 2010 by Taylor and Francis Group, LLC 354 Chlorinated Fatty Acid Medium Chlorinated Fatty Acid Medium Composition per 1001.0mL: (NH 4 ) 2 SO 4 0.5g MgSO 4 ·7H 2 O 0.1g Ca(NO 3 ) 2 0.075g 2-Chloropropionate 0.54g Phosphate buffer (20mM solution, pH 7.2) 1.0L Trace elements solution 1.0mL pH 7.2 ± 0.2 at 25°C Trace Elements Solution: Composition per liter: FeSO 4 ·7H 2 O 1.0g MnSO 4 ·H 2 O 1.0g Co(NO 3 ) 2 ·6H 2 O 0.25g CuCl 2 ·2H 2 O 0.25g Na 2 MoO 4 ·2H 2 O 0.25g ZnCl 2 0.25g H 3 BO 3 0.1g NH 4 VO 3 0.1g NiSO 4 ·6H 2 O 0.1g Preparation of Trace Elements Solution: Add components to distilled/deionized water and bring volume to 1.0L. 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 Alcaligenes species. Chlorobiaceae Medium 1 Composition per 4990.0mL: Solution 1 4.0L O 2 -free water 860.0mL NaHCO 3 solution 100.0mL Na 2 S·9H 2 O solution 20.0mL Trace elements solution 5.0mL Vitamin B 12 solution 5.0mL pH 6.8 ± 0.2 at 25°C Solution 1: Composition per 4.0L: MgSO 4 ·7H 2 O 2.5g KCl 1.7g KH 2 PO 4 1.7g NH 4 Cl 1.7g CaCl 2 ·2H 2 O 1.25g Preparation of Solution 1: Add components to distilled/deionized water and bring volume to 4.0L. Mix thoroughly. Autoclave for 45 min at 15 psi pressure–121°C. Cool to 25°C under 100% N 2 . Saturate with CO 2 by stirring under 100% CO 2 for 30 min. O 2 -Free Water: Composition per 860.0mL: H 2 O 860.0mL Preparation of O 2 -Free Water: Autoclave H 2 O for 15 min at 15 psi pressure–121°C. Cool to 25°C under 100% N 2 . NaHCO 3 Solution: Composition per 100.0mL: NaHCO 3 7.5g Preparation of NaHCO 3 Solution: Add the NaHCO 3 to distilled/ deionized water and bring volume to 100.0mL. Mix thoroughly. Gas with 100% CO 2 for 20 min. Filter sterilize with positive CO 2 pressure. 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. Mix thoroughly. Gas with 100% N 2 for 15 min in a screw-capped bottle. Tightly close cap. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Trace Elements Solution: Composition per liter: FeCl 2 ·4H 2 O 1.5g CoCl 2 ·6H 2 O 0.19g MnCl 2 ·4H 2 O 0.1g ZnCl 2 0.07g H 3 BO 3 0.06g NaMoO 4 ·2H 2 O 0.04g CuCl 2 ·2H 2 O 0.02g NiCl 2 ·6H 2 0 0.02g HCl (25% solution) 6.5mL Preparation of Trace Elements Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Vitamin B 12 Solution: Composition per 100.0mL: Vitamin B 12 2.0mg Preparation of Vitamin B 12 Solution: Add vitamin B 12 to dis- tilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Preparation of Medium: To 4.0L of sterile, CO 2 -saturated solution 1, aseptically add the remaining components. Mix thoroughly. Adjust pH to 6.8. Aseptically distribute into sterile 100.0mL bottles using pos- itive pressure of 95% N 2 + 5% CO 2 . Completely fill bottles with medi- um except for a pea-sized air bubble. Use: For the isolation and cultivation of members of the Chloro- biaceae. Chlorobiaceae Medium 2 Composition per 1051.0mL: Solution 1 950.0mL Na 2 S·9H 2 O solution 60.0mL NaHCO 3 solution 40.0mL Vitamin B 12 solution 1.0mL pH 6.8 ± 0.2 at 25°C Solution 1: Composition per 950.0mL: KH 2 PO 4 1.0g NH 4 Cl 0.5g MgSO 4 ·7H 2 O 0.4g CaCl 2 ·2H 2 O 0.05g Trace elements solution SL-8 1.0mL Preparation of Solution 1: Add components to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. © 2010 by Taylor and Francis Group, LLC . isolation of various bacteria; with the addition of blood, for the cultivation of fastidious bacteria. For the cul- tivation of Bordetella pertussis for vaccine production and the mainte- nance of stock. isolation of various bacteria; with the addition of blood, for the cultivation of fastidious bacteria. For the cul- tivation of Bordetella pertussis for vaccine production and the mainte- nance of stock. cultivation of Brevibacterium linens. Cherry Agar, CBS Formula Composition per liter: Pulp of sour stone cherries 200.0g Agar 20.0g pH 3.8 ± 0.2 at 25°C Preparation of Medium: Add 1.0L of distilled/deionized