Chopped Meat Broth 365 Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Continue boiling for 2–3 min. Allow precipitate to settle out. Distribute supernatant into 125.0mL flasks in 10.0mL volumes. Add standard solution or test solutions to each flask. Adjust the volume of each flask to 20.0mL with distilled/deionized water. Autoclave for 10 min at 15 psi pressure–121°C. Use: For the microbiological assay of choline using Neurospora crassa as the test microorganism. Choline Medium Composition per liter: NaCl 30.0g Choline chloride 5.0g K 2 HPO 4 1.0g MgSO 4 0.5g FeSO 4 0.01g pH 7.4 ± 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 7.4. Dis- tribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure– 121°C. Use: For the cultivation of Achromobacter holinophagum. Chondromyces VYZ Agar Composition per liter: Agar 15.0g Fresh baker’s yeast cake 5.0g CaCl 2 1.0g pH 7.2 ± 0.2 at 25°C Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.2. Dis- tribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure– 121°C. Use: For the cultivation of Chondromyces species. Chopped Liver Broth Composition per liter: Fresh beef liver 500.0g Peptone 10.0g K 2 HPO 4 1.0g Soluble starch 1.0g pH 7.0 ± 0.2 at 25°C Source: This medium is available as a premixed powder from Hi- Media. Preparation of Medium: Grind fresh beef liver. Add to 1.0L of dis- tilled/deionized water. Gently heat and bring to boiling. Continue boil- ing for 60 min. Cool to 25°C. Adjust pH to 7.0. Gently heat and bring to boiling. Continue boiling for 10 min. Filter through cheesecloth. Save chopped liver particles. To filtrate, add remaining components. Bring volume to 1.0L with distilled/deionized water. Adjust pH to 7.0. Filter through Whatman #1 filter paper. Add chopped liver particles to test tubes to a depth of 1.2–2.5 cm. Add 10.0mL of broth to each tube. Autoclave for 15 min at 15 psi pressure–121°C. Use: For the isolation and cultivation of Clostridium botulinum, Clostridium perfringens, and other anaerobic bacteria from foods. Chopped Liver HiVeg Broth Composition per liter: Plant infusion 100.0g Plant peptone 10.0g K 2 HPO 4 1.0g Starch, soluble 1.0g 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 20 min at 15 psi pressure–121°C. Use: For the isolation and cultivation of Clostridium botulinum, Clostridium perfringens, and other anaerobic bacteria from foods. Chopped Meat Agar Composition per liter: Ground meat, fat free 500.0g Pancreatic digest of casein 30.0g Agar 15.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl 0.5g Resazurin 1.0mg NaOH (1N solution) 25.0mL pH 7.0 ± 0.2 at 25°C Preparation of Medium: Use lean beef or horse meat. Remove fat and connective tissue. Grind finely. Add ground meat and 25.0mL of NaOH solution to distilled/deionized water and bring volume to 1025.0mL. Gently heat and bring to boiling. Continue boiling for 15 min without stirring. Cool to room temperature. Remove fat from sur- face. Filter and retain both meat particles and filtrate. Adjust volume of filtrate to 1.0L with distilled/deionized water. Add pancreatic digest of casein, agar, K 2 HPO 4 , yeast extract, and resazurin. Gently heat and bring to boiling. Boil for 1–2 min. Add L-cysteine·HCl. Mix thorough- ly. Distribute 7.0mL into tubes that contain meat particles (1 part meat particles to 5 parts fluid). Autoclave for 30 min at 15 psi pressure– 121°C. Use: For the cultivation of various anaerobes. Chopped Meat Broth Composition per liter: Ground meat, fat free 500.0g Pancreatic digest of casein 30.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl 0.5g Resazurin 1.0mg NaOH (1N solution) 25.0mL pH 7.0 ± 0.2 at 25°C Preparation of Medium: Use lean beef or horse meat. Remove fat and connective tissue. Grind finely. Add ground meat and 25.0mL of NaOH solution to distilled/deionized water and bring volume to 1025.0mL. Gently heat and bring to boiling. Continue boiling for 15 min without stirring. Cool to room temperature. Remove fat from sur- face. Filter and retain both meat particles and filtrate. Adjust volume of filtrate to 1.0L with distilled/deionized water. Add pancreatic digest of casein, K 2 HPO 4 , yeast extract, and resazurin. Gently heat and bring to © 2010 by Taylor and Francis Group, LLC 366 Chopped Meat Broth with Carbohydrates boiling. Boil for 1–2 min. Add L-cysteine·HCl. Mix thoroughly. Dis- tribute 7.0mL into tubes that contain meat particles (1 part meat parti- cles to 5 parts fluid). Autoclave for 30 min at 15 psi pressure–121°C. Use: For the cultivation of various anaerobes. Chopped Meat Broth with Carbohydrates (DSMZ Medium 110) Composition per liter: Ground meat, fat free 500.0g Pancreatic digest of casein 30.0g K 2 HPO 4 5.0g Yeast extract 5.0g Glucose 4.0g Cellobiose 1.0g Maltose 1.0g Starch, soluble 1.0g L-Cysteine·HCl 0.5g Resazurin 1.0mg NaOH (1N solution) 25.0mL pH 7.0 ± 0.2 at 25°C Preparation of Medium: Use lean beef or horse meat. Remove fat and connective tissue. Grind finely. Add ground meat and 25.0mL of NaOH solution to distilled/deionized water and bring volume to 1025.0mL. Gently heat and bring to boiling. Continue boiling for 15 min without stirring. Cool to room temperature. Remove fat from sur- face. Filter and retain both meat particles and filtrate. Adjust volume of filtrate to 1.0L with distilled/deionized water. Add pancreatic digest of casein, K 2 HPO 4 , yeast extract, and resazurin. Gently heat and bring to boiling. Boil for 1–2 min. Add glucose, cellobiose, maltose, and solu- ble starch. Add L-cysteine·HCl. Mix thoroughly. Distribute 7.0mL into tubes that contain meat particles (1 part meat particles to 5 parts fluid). Autoclave for 30 min at 15 psi pressure–121°C. Use: For the cultivation of numerous anaerobes, including Actinomy- ces suis, Anaerobiospirillum succiniciproducens, Bacteroides distaso- nis, Bacteroides eggerthii, Bacteroides fragilis, Bacteroides helco- genes, Bacteroides macacae, Bacteroides pyogenes, Bacteroides splanchnicus, Bacteroides suis, Centipeda periodontii, numerous Clostridium species, Eubacterium brachy, Eubacterium eligens, Eubacterium hallii, Eubacterium limosum, Eubacterium plautii, Eubacterium ruminantium, Eubacterium saburreum, Eubacterium sir- aeum, Eubacterium species, Eubacterium tarantellus, Eubacterium tenue, Eubacterium ventriosum, Megamonas hypermegas, Peptococ- cus niger, Peptostreptococcus productus, Prevotella buccae, Pre- votella buccalis, Prevotella denticola, Prevotella intermedia, Pre- votella oralis, and Selenomonas sputigena. Chopped Meat Broth with Formate and Fumarate (LMG Medium 69) Composition per liter: Ground meat, fat free 500.0g Pancreatic digest of casein 30.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl 0.5g Resazurin 1.0mg NaOH (1N solution) 25.0mL Formate-fumarate solution 7.7mL pH 7.0 ± 0.2 at 25°C Formate-Fumarate Solution: Composition per 100.0mL: Sodium formate 6.0g Fumaric acid 6.0g Preparation of Formate-Fumarate Solution: Add components to distilled/deionized water and bring volume to 100.0mL. Adjust pH to 7.0. Filter sterilize. Preparation of Medium: Use lean beef or horse meat. Remove fat and connective tissue. Grind finely. Add ground meat and 25.0mL of NaOH solution to distilled/deionized water and bring volume to 1025.0mL. Gently heat and bring to boiling. Continue boiling for 15 min without stirring. Cool to room temperature. Remove fat from sur- face. Filter and retain both meat particles and filtrate. Adjust volume of filtrate to 1.0L with distilled/deionized water. Add pancreatic digest of casein, agar, K 2 HPO 4 , yeast extract, and resazurin. Gently heat and bring to boiling. Boil for 1–2 min. Add L-cysteine·HCl. Mix thorough- ly. Distribute 6.5mL into tubes that contain meat particles (1 part meat particles to 5 parts fluid). Autoclave for 30 min at 15 psi pressure– 121°C. Cool to 25°C. Aseptically add 50 µL of sterile formate/fumar- ate solution to each tube prior to inoculation. Use: For the cultivation of various Clostridium spp. Chopped Meat Broth with Vitamin K 1 (LMG Medium 70) Composition per liter: Ground meat, fat free 500.0g Pancreatic digest of casein 30.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl 0.5g Resazurin 1.0mg NaOH (1N solution) 25.0mL Vitamin K 1 solution 7.7mL pH 7.0 ± 0.2 at 25°C Vitamin K 1 Solution: Composition per 50.0mL: Ethanol (20% solution) 50.0mL Vitamin K 1 0.7gL Preparation of Vitamin K 1 Solution: Mix components. Filter sterilize. Store solution protected from light at 5°C. Discard after one month. Preparation of Medium: Use lean beef or horse meat. Remove fat and connective tissue. Grind finely. Add ground meat and 25.0mL of NaOH solution to distilled/deionized water and bring volume to 1025.0mL. Gently heat and bring to boiling. Continue boiling for 15 min without stirring. Cool to room temperature. Remove fat from sur- face. Filter and retain both meat particles and filtrate. Adjust volume of filtrate to 1.0L with distilled/deionized water. Add pancreatic digest of casein, agar, K 2 HPO 4 , yeast extract, and resazurin. Gently heat and bring to boiling. Boil for 1–2 min. Add L-cysteine·HCl. Mix thorough- ly. Distribute 6.5mL into tubes that contain meat particles (1 part meat particles to 5 parts fluid). Autoclave for 30 min at 15 psi pressure– 121°C. Cool to 25°C. Aseptically add 50 µL of sterile vitamin K 1 solu- tion to each tube prior to inoculation. Use: For the cultivation of various Clostridium spp. © 2010 by Taylor and Francis Group, LLC Chopped Meat Carbohydrate Medium with Tween™ 80 367 Chopped Meat Carbohydrate Medium Composition per 1240.0mL: Peptone 30.0g K 2 HPO 4 5.0g Yeast extract 5.0g Cellobiose 1.0g Maltose 1.0g Starch 1.0g L-Cysteine·HCl·H 2 O 0.5g Chopped meat extract filtrate 1.0L Chopped meat extract solids 200.0mL Resazurin (0.025% solution) 4.0mL pH 7.0 ± 0.2 at 25°C Chopped Meat Extract: Composition per liter: Beef or horse meat 500.0g NaOH (1N solution) 25.0mL Preparation of Chopped Meat Extract: Use lean beef or horse meat. Remove fat and connective tissue. Grind. Add meat and NaOH to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling while stirring. Cool to 25°C. Remove fat from surface. Filter. Reserve ground meat particles and filtrate. Add distilled/deion- ized water to filtrate and bring volume to 1.0L. Preparation of Medium: To 1.0L of chopped meat extract filtrate, add the remaining components, except the L-cysteine·HCl·H 2 O and chopped meat solids. Mix thoroughly. Gently heat to boiling. Cool to room temperature. Add the L-cysteine·HCl·H 2 O. Adjust pH to 7.0. Dis- tribute 1 part chopped meat solids (by volume) and 5 parts of liquid (by volume) into tubes under O 2 -free 97% N 2 + 3% H 2 . Cap with rubber stoppers and place tubes in a press. Autoclave for 15 min at 15 psi pres- sure–121°C with fast exhaust. Use: For the cultivation of anaerobic bacteria, including Clostridium species, Eubacterium species, and Gemmiger formicilis. Chopped Meat Carbohydrate Medium with Rumen Fluid (ATCC Medium 1016) Composition per 1390.0mL: Peptone 30.0g K 2 HPO 4 5.0g Yeast extract 5.0g Cellobiose 1.0g Maltose 1.0g Starch 1.0g L-Cysteine·HCl·H 2 O 0.5g Chopped meat extract filtrate 1.0L Chopped meat extract solids 200.0mL Rumen fluid 150.0mL Resazurin (0.025% solution) 4.0mL pH 7.0 ± 0.2 at 25°C Chopped Meat Extract: Composition per liter: Beef or horse meat 500.0g NaOH (1N solution) 25.0mL Preparation of Chopped Meat Extract: Use lean beef or horse meat. Remove fat and connective tissue. Grind. Add meat and NaOH to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling while stirring. Cool to 25°C. Remove fat from surface. Filter. Reserve ground meat particles and filtrate. Add distilled/deion- ized water to filtrate and bring volume to 1.0L. Preparation of Medium: To 1.0L of chopped meat extract filtrate, add the remaining components, except L-cysteine·HCl·H 2 O and chopped meat solids. Mix thoroughly. Gently heat to boiling. Cool to room temperature. Add the L-cysteine·HCl·H 2 O. Adjust pH to 7.0. Dis- tribute 1 part chopped meat solids (by volume) and 5 parts of liquid (by volume) into tubes under O 2 -free 97% N 2 + 3% H 2 . Cap with rubber stoppers and place tubes in a press. Autoclave for 15 min at 15 psi pres- sure–121°C with fast exhaust. Use: For the cultivation of anaerobic bacteria, including Butyrivibrio crossotus, Eubacterium species, and Ruminococcus species. Chopped Meat Carbohydrate Medium with Rumen Fluid Composition per 1390.0mL: Peptone 30.0g K 2 HPO 4 5.0g Yeast extract 5.0g Glucose 4.0g Cellobiose 1.0g Maltose 1.0g Starch 1.0g L-Cysteine·HCl·H 2 O 0.5g Chopped meat extract filtrate 1.0L Chopped meat extract solids 200.0mL Rumen fluid 150.0mL Resazurin (0.025% solution) 4.0mL pH 7.0 ± 0.2 at 25°C Chopped Meat Extract: Composition per liter: Beef or horse meat 500.0g NaOH (1N solution) 25.0mL Preparation of Chopped Meat Extract: Use lean beef or horse meat. Remove fat and connective tissue. Grind. Add meat and NaOH to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling while stirring. Cool to 25°C. Remove fat from surface. Filter. Reserve ground meat particles and filtrate. Add distilled/deion- ized water to filtrate and bring volume to 1.0L. Preparation of Medium: To 1.0L of chopped meat extract filtrate, add the remaining components, except L-cysteine·HCl·H 2 O and chopped meat solids. Mix thoroughly. Gently heat to boiling. Cool to room temperature. Add the L-cysteine·HCl·H 2 O. Adjust pH to 7.0. Dis- tribute 1 part chopped meat solids (by volume) and 5 parts of liquid (by volume) into tubes under O 2 -free 97% N 2 + 3% H 2 . Cap with rubber stoppers and place tubes in a press. Autoclave for 15 min at 15 psi pres- sure–121°C with fast exhaust. Use: For the cultivation of anaerobic bacteria, including Fusobacte- rium prausnitzii, Eubacterium species, and Prevotella ruminicola. Chopped Meat Carbohydrate Medium with Tween™ 80 Composition per 1240.0mL: Peptone 30.0g K 2 HPO 4 5.0g Yeast extract 5.0g Cellobiose 1.0g Maltose 1.0g Starch 1.0g © 2010 by Taylor and Francis Group, LLC 368 Chopped Meat Glucose Agar Tween™ 80 1.0g L-Cysteine·HCl·H 2 O 0.5g Chopped meat extract filtrate 1.0L Chopped meat extract solids 200.0mL Resazurin (0.025% solution) 4.0mL pH 7.0 ± 0.2 at 25°C Chopped Meat Extract: Composition per liter: Beef or horse meat 500.0g NaOH (1N solution) 25.0mL Preparation of Chopped Meat Extract: Use lean beef or horse meat. Remove fat and connective tissue. Grind. Add meat and NaOH to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling while stirring. Cool to 25°C. Remove fat from surface. Filter. Reserve ground meat particles and filtrate. Add distilled/deion- ized water to filtrate and bring volume to 1.0L. Preparation of Medium: To 1.0L of chopped meat extract filtrate, add the remaining components, except L-cysteine·HCl·H 2 O and chopped meat solids. Mix thoroughly. Gently heat to boiling. Cool to room temperature. Add the L-cysteine·HCl·H 2 O. Adjust pH to 7.0. Dis- tribute 1 part chopped meat solids (by volume) and 5 parts of liquid (by volume) into tubes under O 2 -free 97% N 2 + 3% H 2 . Cap with rubber stoppers and place tubes in a press. Autoclave for 15 min at 15 psi pres- sure–121°C with fast exhaust. Use: For the cultivation of Coprococcus species and Peptostreptococ- cus micros. Chopped Meat Glucose Agar (LMG Medium 68) Composition per liter: Ground meat, fat free 500.0g Pancreatic digest of casein 30.0g Agar 15.0g Glucose 10.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl 0.5g Resazurin 1.0mg NaOH (1N solution) 25.0mL pH 7.0 ± 0.2 at 25°C Preparation of Medium: Use lean beef or horse meat. Remove fat and connective tissue. Grind finely. Add ground meat and 25.0mL of NaOH solution to distilled/deionized water and bring volume to 1025.0mL. Gently heat and bring to boiling. Continue boiling for 15 min without stirring. Cool to room temperature. Remove fat from sur- face. Filter and retain both meat particles and filtrate. Adjust volume of filtrate to 1.0L with distilled/deionized water. Add pancreatic digest of casein, agar, K 2 HPO 4 , yeast extract, and resazurin. Gently heat and bring to boiling. Boil for 1–2 min. Add L-cysteine·HCl. Mix thorough- ly. Distribute 7.0mL into tubes that contain meat particles (1 part meat particles to 5 parts fluid). Autoclave for 30 min at 15 psi pressure– 121°C. Use: For the cultivation of various Clostridium spp. Chopped Meat Glucose Broth (LMG Medium 68) Composition per liter: Ground meat, fat free 500.0g Pancreatic digest of casein 30.0g Glucose 10.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl 0.5g Resazurin 1.0mg NaOH (1N solution) 25.0mL pH 7.0 ± 0.2 at 25°C Preparation of Medium: Use lean beef or horse meat. Remove fat and connective tissue. Grind finely. Add ground meat and 25.0mL of NaOH solution to distilled/deionized water and bring volume to 1025mL. Gently heat and bring to boiling. Continue boiling for 15 min without stirring. Cool to room temperature. Remove fat from surface. Filter and retain both meat particles and filtrate. Adjust volume of fil- trate to 1.0L with distilled/deionized water. Add pancreatic digest of casein, agar, K 2 HPO 4 , yeast extract, and resazurin. Gently heat and bring to boiling. Boil for 1–2 min. Add L-cysteine·HCl. Mix thorough- ly. Distribute 7.0mL into tubes that contain meat particles (1 part meat particles to 5 parts fluid). Autoclave for 30 min at 15 psi pressure– 121°C. Use: For the cultivation of various Clostridium spp. Chopped Meat Glucose Medium Composition per 1240.0mL: Peptone 30.0g K 2 HPO 4 5.0g Yeast extract 5.0g Glucose 5.0g L-Cysteine·HCl·H 2 O 0.5g Chopped meat extract filtrate 1.0L Chopped meat extract solids 200.0mL Resazurin (0.025% solution) 4.0mL pH 7.0 ± 0.2 at 25°C Chopped Meat Extract: Composition per liter: Beef or horse meat 500.0g NaOH (1N solution) 25.0mL Preparation of Chopped Meat Extract: Use lean beef or horse meat. Remove fat and connective tissue. Grind. Add meat and NaOH to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling while stirring. Cool to 25°C. Remove fat from surface. Filter. Reserve ground meat particles and filtrate. Add distilled/deion- ized water to filtrate and bring volume to 1.0L. Preparation of Medium: To 1.0L of chopped meat extract filtrate, add the remaining components, except L-cysteine·HCl·H 2 O and chopped meat solids. Mix thoroughly. Gently heat to boiling. Cool to room temperature. Add the L-cysteine·HCl·H 2 O. Adjust pH to 7.0. Dis- tribute 1 part chopped meat solids (by volume) and 5 parts of liquid (by volume) into tubes under O 2 -free 97% N 2 + 3% H 2 . Cap with rubber stoppers and place tubes in a press. Autoclave for 15 min at 15 psi pres- sure–121°C with fast exhaust. Use: For the cultivation of Clostridium species and Selenomonas noxia. Chopped Meat Glucose Medium with NaCl Composition per 1205.0mL: NaCl 30.0g Peptone 30.0g K 2 HPO 4 5.0g Yeast extract 5.0g © 2010 by Taylor and Francis Group, LLC Chopped Meat Medium with Formate and Fumarate 369 Glucose 5.0g L-Cysteine·HCl·H 2 O 0.5g Chopped meat extract filtrate 1.0L Chopped meat extract solids 200.0mL Resazurin (0.025% solution) 4.0mL pH 7.0 ± 0.2 at 25°C Chopped Meat Extract: Composition per liter: Beef or horse meat 500.0g NaOH (1N solution) 25.0mL Preparation of Chopped Meat Extract: Use lean beef or horse meat. Remove fat and connective tissue. Grind. Add meat and NaOH to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling while stirring. Cool to 25°C. Remove fat from surface. Filter. Reserve ground meat particles and filtrate. Add distilled/deion- ized water to filtrate and bring volume to 1.0L. Preparation of Medium: To 1.0L of chopped meat extract filtrate, add the remaining components, except the L-cysteine·HCl·H 2 O and chopped meat solids. Mix thoroughly. Gently heat to boiling. Cool to room temperature. Add the L-cysteine·HCl·H 2 O. Adjust pH to 7.0. Dis- tribute 1 part chopped meat solids (by volume) and 5 parts of liquid (by volume) into tubes under O 2 -free 97% N 2 + 3% H 2 . Cap with rubber stoppers and place tubes in a press. Autoclave for 15 min at 15 psi pres- sure–121°C with fast exhaust. Use: For the cultivation and maintenance of anaerobic halophilic bac- teria. Chopped Meat Medium Composition per 1205.0mL: Peptone 30.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl·H 2 O 0.5g Chopped meat extract filtrate 1.0L Chopped meat extract solids 200.0mL Resazurin (0.025% solution) 4.0mL pH 7.0 ± 0.2 at 25°C Chopped Meat Extract: Composition per liter: Beef or horse meat 500.0g NaOH (1N solution) 25.0mL Preparation of Chopped Meat Extract: Use lean beef or horse meat. Remove fat and connective tissue. Grind. Add meat and NaOH to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling while stirring. Cool to 25°C. Remove fat from surface. Filter. Reserve ground meat particles and filtrate. Add distilled/deion- ized water to filtrate and bring volume to 1.0L. Preparation of Medium: To 1.0L of chopped meat extract filtrate, add the remaining components, except the L-cysteine·HCl·H 2 O and chopped meat solids. Mix thoroughly. Gently heat to boiling. Cool to room temperature. Add the L-cysteine·HCl·H 2 O. Adjust pH to 7.0. Dis- tribute 1 part chopped meat solids (by volume) and 5 parts of liquid (by volume) into tubes under O 2 -free 97% N 2 + 3% H 2 . Cap with rubber stoppers and place tubes in a press. Autoclave for 15 min at 15 psi pres- sure–121°C with fast exhaust. Use: For the cultivation and maintenance of a variety of anaerobic bacteria, including Bacteroides species, Bifidobacterium species, Cap- nocytophaga species, Clostridium species, Eubacterium species, Fuso- bacterium species, Peptostreptococcus species, Prevotella species, Propionibacterium species, Ruminococcus species, and others. Chopped Meat Medium with 10% Fetal Calf Serum Composition per 1230.0mL: Peptone 30.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl·H 2 O 0.5g Chopped meat extract filtrate 1.0L Chopped meat extract solids 200.0mL Fetal calf serum 100.0mL Resazurin (0.025% solution) 4.0mL pH 7.0 ± 0.2 at 25°C Chopped Meat Extract: Composition per liter: Beef or horse meat 500.0g NaOH (1N solution) 25.0mL Preparation of Chopped Meat Extract: Use lean beef or horse meat. Remove fat and connective tissue. Grind. Add meat and NaOH to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling while stirring. Cool to 25°C. Remove fat from surface. Filter. Reserve ground meat particles and filtrate. Add distilled/deion- ized water to filtrate and bring volume to 1.0L. Preparation of Medium: To 1.0L of chopped meat extract filtrate, add the remaining components, except the L-cysteine·HCl·H 2 O and chopped meat solids. Mix thoroughly. Gently heat to boiling. Cool to room temperature. Add the L-cysteine·HCl·H 2 O. Adjust pH to 7.0. Dis- tribute 1 part chopped meat solids (by volume) and 5 parts of liquid (by volume) into tubes under O 2 -free 97% N 2 + 3% H 2 . Cap with rubber stoppers and place tubes in a press. Autoclave for 15 min at 15 psi pres- sure–121°C with fast exhaust. Use: For the cultivation and maintenance of Actinomyces hordeovul- neris. Chopped Meat Medium with Formate and Fumarate Composition per 1230.0mL: Peptone 30.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl·H 2 O 0.5g Chopped meat extract filtrate 1.0L Chopped meat extract solids 200.0mL Resazurin (0.025% solution) 4.0mL Formate-fumarate solution 0.05mL pH 7.0 ± 0.2 at 25°C Chopped Meat Extract: Composition per liter: Beef or horse meat 500.0g NaOH (1N solution) 25.0mL Preparation of Chopped Meat Extract: Use lean beef or horse meat. Remove fat and connective tissue. Grind. Add meat and NaOH to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling while stirring. Cool to 25°C. Remove fat from surface. Filter. Reserve ground meat particles and filtrate. Add distilled/deion- ized water to filtrate and bring volume to 1.0L. © 2010 by Taylor and Francis Group, LLC 370 Chopped Meat Medium with 1% Glucose Formate-Fumarate Solution: Composition per 100.0mL: Sodium formate 6.0g Fumaric acid 6.0g Preparation of Formate-Fumarate Solution: Add components to distilled/deionized water and bring volume to 100.0mL. Adjust pH to 7.0. Filter sterilize. Preparation of Medium: To 1.0L of chopped meat extract filtrate, add the remaining components, except the L-cysteine·HCl·H 2 O, formate- fumarate solution, and chopped meat solids. Mix thoroughly. Gently heat to boiling. Cool to room temperature. Add the L-cysteine·HCl·H 2 O. Ad- just pH to 7.0. Distribute 1 part chopped meat solids (by volume) and 5 parts of liquid (by volume) into tubes under O 2 -free 97% N 2 + 3% H 2 . Cap with rubber stoppers and place tubes in a press. Autoclave for 15 min at 15 psi pressure–121°C with fast exhaust. Prior to inoculation, add 0.05mL of formate-fumarate solution to each tube containing approxi- mately 6.5mL of chopped meat medium. Use: For the cultivation and maintenance of Bacteroides ureolyticus and Wolinella species. Chopped Meat Medium with 1% Glucose Composition per 1230.0mL: Peptone 30.0g Glucose 10.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl·H 2 O 0.5g Chopped meat extract filtrate 1.0L Chopped meat extract solids 200.0mL Resazurin (0.025% solution) 4.0mL pH 7.0 ± 0.2 at 25°C Chopped Meat Extract: Composition per liter: Beef or horse meat 500.0g NaOH (1N solution) 25.0mL Preparation of Chopped Meat Extract: Use lean beef or horse meat. Remove fat and connective tissue. Grind. Add meat and NaOH to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling while stirring. Cool to 25°C. Remove fat from surface. Filter. Reserve ground meat particles and filtrate. Add distilled/deion- ized water to filtrate and bring volume to 1.0L. Preparation of Medium: To 1.0L of chopped meat extract filtrate, add the remaining components, except the L-cysteine·HCl·H 2 O and chopped meat solids. Mix thoroughly. Gently heat to boiling. Cool to room temperature. Add the L-cysteine. Adjust pH to 7.0. Distribute 1 part chopped meat solids (by volume) and 5 parts of liquid (by volume) into tubes under O 2 -free 97% N 2 + 3% H 2 . Cap with rubber stoppers and place tubes in a press. Autoclave for 15 min at 15 psi pressure– 121°C with fast exhaust. Use: For the cultivation and maintenance of anaerobic bacteria, including Bacteroides disiens, Coprococcus eutastus, Eubacterium formicigenerans, Prevotella disiens, Ruminococcus torques, and Streptococcus hansenii. Chopped Meat Medium with Menadione Composition per 1230.0mL: Peptone 30.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl·H 2 O 0.5g Chopped meat extract filtrate 1.0L Chopped meat extract solids 200.0mL Resazurin (0.025% solution) 4.0mL Menadione solution 0.25mL pH 7.0 ± 0.2 at 25°C Chopped Meat Extract: Composition per liter: Beef or horse meat 500.0g NaOH (1N solution) 25.0mL Preparation of Chopped Meat Extract: Use lean beef or horse meat. Remove fat and connective tissue. Grind. Add meat and NaOH to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling while stirring. Cool to 25°C. Remove fat from surface. Filter. Reserve ground meat particles and filtrate. Add distilled/deion- ized water to filtrate and bring volume to 1.0L. Menadione Solution: Composition per liter: Menadione (vitamin K 3 ) 50.0μg Ethanol (20% solution) 25.0mL Preparation of Menadione Solution: Dissolve menadione in eth- anol. Filter sterilize. Preparation of Medium: To 1.0L of chopped meat extract filtrate, add the remaining components, except the L-cysteine·HCl·H 2 O, me- nadione solution, and chopped meat solids. Mix thoroughly. Gently heat to boiling. Cool to room temperature. Add the L- cysteine·HCl·H 2 O. Adjust pH to 7.0. Distribute 1 part chopped meat solids (by volume) and 5 parts of liquid (by volume) into tubes under O 2 -free 97% N 2 + 3% H 2 . Cap with rubber stoppers and place tubes in a press. Autoclave for 15 min at 15 psi pressure–121°C with fast exhaust. Prior to inoculation, add 0.25mL of menadione solution to each tube containing approximately 5.0mL of chopped meat medium. Use: For the cultivation and maintenance of Bacteroides gingivalis, Bacteroides macacae, and Porphyromonas gingivalis. Chopped Meat Medium, Modified (DSMZ Medium 797) Composition per 1230.0mL: Chopped meat extract solids 200.0g Trypticase™ 30.0g Agar 20.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl·H 2 O 0.5g Chopped meat extract filtrate 1.0L Hemin solution 10.0mL Resazurin (0.025% solution) 4.0mL Vitamin K 1 solution 0.2mL pH 7.0 ± 0.2 at 25°C Chopped Meat Extract: Composition per liter: Beef or horse meat 500.0g NaOH (1N solution) 25.0mL Preparation of Chopped Meat Extract: Use lean beef or horse meat. Remove fat and connective tissue. Grind. Add meat and NaOH to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling while stirring. Cool to 25°C. Remove fat from surface. © 2010 by Taylor and Francis Group, LLC Chopped Meat Medium, Modified with Arginine 371 Filter. Reserve both ground meat particles and filtrate. Add distilled/ deionized water to filtrate and bring volume to 1.0L. Hemin Solution: Composition per 100.0mL: Hemin 0.05g NaOH (1N solution) 1.0mL Preparation of Hemin Solution: Add components to distilled/de- ionized water and bring volume to 100.0mL. Mix thoroughly. Vitamin K 1 Solution: Composition per 30.0mL: Ethanol (95% solution) 30.0mL Vitamin K 1 0.15mL Preparation of Vitamin K 1 Solution: Mix components. Store so- lution protected from light at 5°C. Discard after 1 month. Preparation of Medium: To 1.0L of chopped meat extract filtrate, add the remaining components, except the L-cysteine·HCl·H 2 O, hemin solution, vitamin K 1 solution, and chopped meat solids. Mix thor- oughly. Gently heat to boiling. Boil for 5 min. Cool to 25°C while sparging with 80% N 2 + 10% H 2 + 10% CO 2 . Add the L- cysteine·HCl·H 2 O, hemin solution, and vitamin K 1 solution. Adjust pH to 7.0. Distribute 1 part chopped meat solids (by volume) and 5 parts of liquid (by volume) into tubes under 80% N 2 + 10% H 2 + 10% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Use: For the cultivation of Dialister pneumosintes. Chopped Meat Medium, Modified Composition per 1230.0mL: Pancreatic digest of casein 30.0g Peptone 30.0g Agar 20.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl·H 2 O 0.5g Chopped meat extract filtrate 1.0L Chopped meat extract solids 200.0mL Hemin solution 10.0mL Resazurin (0.025% solution) 4.0mL Vitamin K 1 solution 0.2mL pH 7.0 ± 0.2 at 25°C Chopped Meat Extract: Composition per liter: Beef or horse meat 500.0g NaOH (1N solution) 25.0mL Preparation of Chopped Meat Extract: Use lean beef or horse meat. Remove fat and connective tissue. Grind. Add meat and NaOH to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling while stirring. Cool to 25°C. Remove fat from surface. Filter. Reserve ground meat particles and filtrate. Add distilled/deion- ized water to filtrate and bring volume to 1.0L. Hemin Solution: Composition per 100.0mL: Hemin 0.05g NaOH (1N solution) 1.0mL Preparation of Hemin Solution: Add components to distilled/de- ionized water and bring volume to 100.0mL. Mix thoroughly. Vitamin K 1 Solution: Composition per 30.0mL: Ethanol (95% solution) 30.0mL Vitamin K 1 0.15mL Preparation of Vitamin K 1 Solution: Mix components. Store so- lution protected from light at 5°C. Discard after 1 month. Preparation of Medium: To 1.0L of chopped meat extract filtrate, add the remaining components, except the L-cysteine·HCl·H 2 O, hemin solution, vitamin K 1 solution, and chopped meat solids. Mix thorough- ly. Gently heat to boiling. Cool to room temperature. Add the L- cysteine·HCl·H 2 O, hemin solution, and vitamin K 1 solution. Adjust pH to 7.0. Distribute 1 part chopped meat solids (by volume) and 5 parts of liquid (by volume) into tubes under O 2 -free 97% N 2 + 3% H 2 . Cap with rubber stoppers and place tubes in a press. Autoclave for 15 min at 15 psi pressure–121°C with fast exhaust. Use: For the cultivation and maintenance of a variety of anaerobic bacteria, including Actinomyces species, Bacteroides species, Clostrid- ium species, Eubacterium species, Fusobacterium species, Peptostrep- tococcus species, Porphyromonas species, Prevotella species, Propi- onibacterium species, Selenomonas species, and others. Chopped Meat Medium, Modified with Arginine Composition per 1230.0mL: Pancreatic digest of casein 30.0g Peptone 30.0g Agar 20.0g Arginine 5.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl·H 2 O 0.5g Chopped meat extract filtrate 1.0L Chopped meat extract solids 200.0mL Hemin solution 10.0mL Resazurin (0.025% solution) 4.0mL Vitamin K 1 solution 0.2mL pH 7.0 ± 0.2 at 25°C Chopped Meat Extract: Composition per liter: Beef or horse meat 500.0g NaOH (1N solution) 25.0mL Tween™ 80 25.0mL Preparation of Chopped Meat Extract: Use lean beef or horse meat. Remove fat and connective tissue. Grind. Add meat and NaOH to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling while stirring. Cool to 25°C. Remove fat from surface. Filter. Reserve ground meat particles and filtrate. Add distilled/deion- ized water to filtrate and bring volume to 1.0L. Hemin Solution: Composition per 100.0mL: Hemin 0.05g NaOH (1N solution) 1.0mL Preparation of Hemin Solution: Add components to distilled/de- ionized water and bring volume to 100.0mL. Mix thoroughly. Vitamin K 1 Solution: Composition per 30.0mL: Ethanol (95% solution) 30.0mL Vitamin K 1 0.15mL © 2010 by Taylor and Francis Group, LLC 372 Chopped Meat Medium, Modified with Formate and Fumarate Preparation of Vitamin K 1 Solution: Mix components. Store so- lution protected from light at 5°C. Discard after 1 month. Preparation of Medium: To 1.0L of chopped meat extract filtrate, add the remaining components, except the L-cysteine·HCl·H 2 O, hemin solution, vitamin K 1 solution, and chopped meat solids. Mix thorough- ly. Gently heat to boiling. Cool to room temperature. Add the L- cysteine·HCl·H 2 O, hemin solution, and vitamin K 1 solution. Adjust pH to 7.0. Distribute 1 part chopped meat solids (by volume) and 5 parts of liquid (by volume) into tubes under O 2 -free 97% N 2 + 3% H 2 . Cap with rubber stoppers and place tubes in a press. Autoclave for 15 min at 15 psi pressure–121°C with fast exhaust. Use: For the cultivation and maintenance of Eubacterium lentum. Chopped Meat Medium, Modified with Formate and Fumarate Composition per 1230.0mL: Pancreatic digest of casein 30.0g Peptone 30.0g Agar 20.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl·H 2 O 0.5g Chopped meat extract filtrate 1.0L Chopped meat extract solids 200.0mL Hemin solution 10.0mL Resazurin (0.025% solution) 4.0mL Formate-fumarate solution 0.25mL Vitamin K 1 solution 0.2mL pH 7.0 ± 0.2 at 25°C Chopped Meat Extract: Composition per liter: Beef or horse meat 500.0g NaOH (1N solution) 25.0mL Preparation of Chopped Meat Extract: Use lean beef or horse meat. Remove fat and connective tissue. Grind. Add meat and NaOH to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling while stirring. Cool to 25°C. Remove fat from surface. Filter. Reserve ground meat particles and filtrate. Add distilled/deion- ized water to filtrate and bring volume to 1.0L. Hemin Solution: Composition per 100.0mL: Hemin 0.05g NaOH (1N solution) 1.0mL Preparation of Hemin Solution: Add components to distilled/de- ionized water and bring volume to 100.0mL. Mix thoroughly. Formate-Fumarate Solution: Composition per 100.0mL: Sodium formate 6.0g Fumaric acid 6.0g Preparation of Formate-Fumarate Solution: Add components to distilled/deionized water and bring volume to 100.0mL. Adjust pH to 7.0. Filter sterilize. Vitamin K 1 Solution: Composition per 30.0mL: Ethanol (95% solution) 30.0mL Vitamin K 1 0.15mL Preparation of Vitamin K 1 Solution: Mix components. Store so- lution protected from light at 5°C. Discard after one month. Preparation of Medium: To 1.0L of chopped meat extract filtrate, add the remaining components, except the L-cysteine·HCl·H 2 O, hemin solution, vitamin K 1 solution, formate-fumarate solution, and chopped meat solids. Mix thoroughly. Gently heat to boiling. Cool to room tem- perature. Add the L-cysteine·HCl·H 2 O, hemin solution, and vitamin K 1 solution. Adjust pH to 7.0. Distribute 1 part chopped meat solids (by volume) and 5 parts of liquid (by volume) into tubes under O 2 -free 97% N 2 + 3% H 2 . Cap with rubber stoppers and place tubes in a press. Autoclave for 15 min at 15 psi pressure–121°C with fast exhaust. Prior to inoculation, add 0.25mL of formate-fumarate solution to each tube containing approximately 5.0mL of chopped meat medium, modified. Use: For the cultivation and maintenance of Bacteroides gracilis, Bacteroides ureolyticus, Campylobacter mucosalis, and Wolinella suc- cinogenes. Chopped Meat Medium, Modified with Tween™ 80 Composition per 1230.0mL: Pancreatic digest of casein 30.0g Peptone 30.0g Agar 20.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl·H 2 O 0.5g Chopped meat extract filtrate 1.0L Chopped meat extract solids 200.0mL Hemin solution 10.0mL Resazurin (0.025% solution) 4.0mL Vitamin K 1 solution 0.2mL pH 7.0 ± 0.2 at 25°C Chopped Meat Extract: Composition per liter: Beef or horse meat 500.0g NaOH (1N solution) 25.0mL Tween™ 80 25.0mL Preparation of Chopped Meat Extract: Use lean beef or horse meat. Remove fat and connective tissue. Grind. Add meat and NaOH to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling while stirring. Cool to 25°C. Remove fat from surface. Filter. Reserve ground meat particles and filtrate. Add distilled/deion- ized water to filtrate and bring volume to 1.0L. Hemin Solution: Composition per 100.0mL: Hemin 0.05g NaOH (1N solution) 1.0mL Preparation of Hemin Solution: Add components to distilled/de- ionized water and bring volume to 100.0mL. Mix thoroughly. Vitamin K 1 Solution: Composition per 30.0mL: Ethanol (95% solution) 30.0mL Vitamin K 1 0.15mL Preparation of Vitamin K 1 Solution: Mix components. Store so- lution protected from light at 5°C. Discard after 1 month. Preparation of Medium: To 1.0L of chopped meat extract filtrate, add the remaining components, except the L-cysteine·HCl·H 2 O, hemin solution, vitamin K 1 solution, and chopped meat solids. Mix thoroughly. Gently heat to boiling. Cool to room temperature. Add the L- © 2010 by Taylor and Francis Group, LLC Chopped Meat Medium with 0.025% Tween™ 80 373 cysteine·HCl·H 2 O, hemin solution, and vitamin K 1 solution. Adjust pH to 7.0. Distribute 1 part chopped meat solids (by volume) and 5 parts of liquid (by volume) into tubes under O 2 -free 97% N 2 + 3% H 2 . Cap with rubber stoppers and place tubes in a press. Autoclave for 15 min at 15 psi pressure–121°C with fast exhaust. Use: For the cultivation and maintenance of Lactobacillus species and Eubacterium biforme. Chopped Meat Medium with 10% Reduced Filtered Rumen Fluid Composition per 1330.0mL: Peptone 30.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl·H 2 O 0.5g Chopped meat extract filtrate 1.0L Chopped meat extract solids 200.0mL Rumen fluid, reduced and filtered 100.0mL Resazurin (0.025% solution) 4.0mL pH 7.0 ± 0.2 at 25°C Chopped Meat Extract: Composition per liter: Beef or horse meat 500.0g NaOH (1N solution) 25.0mL Preparation of Chopped Meat Extract: Use lean beef or horse meat. Remove fat and connective tissue. Grind. Add meat and NaOH to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling while stirring. Cool to 25°C. Remove fat from surface. Filter. Reserve ground meat particles and filtrate. Add distilled/deion- ized water to filtrate and bring volume to 1.0L. Preparation of Medium: To 1.0L of chopped meat extract filtrate, add the remaining components, except the L-cysteine·HCl·H 2 O and chopped meat solids. Mix thoroughly. Gently heat to boiling. Cool to room temperature. Add the L-cysteine·HCl·H 2 O. Adjust pH to 7.0. Dis- tribute 1 part chopped meat solids (by volume) and 5 parts of liquid (by volume) into tubes under O 2 -free 97% N 2 + 3% H 2 . Cap with rubber stoppers and place tubes in a press. Autoclave for 15 min at 15 psi pres- sure–121°C Use: For the cultivation and maintenance of Eubacterium hallii. Chopped Meat Medium for Treponema spp. (DSMZ Medium 78a) Composition per 1055.0mL: Ground meat, fat free 500.0g Pancreatic digest of casein 30.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl 0.5g Resazurin 1.0mg Amino acid solution 50.0mL NaOH (1N solution) 25.0mL Vitamin solution 5.0mL pH 7.0 ± 0.2 at 25°C Amino Acid Solution: Composition per liter: L-Glutamine 0.7g L-Histidine 0.6g L-Serine 0.5g Preparation of Amino Acid Solution: Add components to dis- tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Vitamin Solution: Composition per liter: Vitamin B 12 50.0mg Pantothenic acid 50.0mg Riboflavin 50.0mg α-Lipoic acid 50.0mg p-Aminobenzoic acid 50.0mg Thiamine-HCl·2H 2 O 50.0mg Nicotinic acid 25.0mg Nicotine amide 25.0mg Biotin 20.0mg Folic acid 20.0mg Pyridoxamine-HCl 10.0mg Preparation of Vitamin Solution: Add components to distilled/ deionized water and bring volume to 1.0L. Mix thoroughly. Filter ster- ilize. Preparation of Medium: Use lean beef or horse meat. Remove fat and connective tissue. Grind finely. Add ground meat and 25.0mL of NaOH solution to distilled/deionized water and bring volume to 1025.0mL. Gently heat and bring to boiling. Continue boiling for 15 min without stirring. Cool to room temperature. Remove fat from sur- face. Filter and retain both meat particles and filtrate. Adjust volume of filtrate to 1.0L with distilled/deionized water. Add pancreatic digest of casein, K 2 HPO 4 , yeast extract, and resazurin. Gently heat and bring to boiling. Boil for 1–2 min. Add L-cysteine·HCl. Mix thoroughly. Auto- clave for 30 min at 15 psi pressure–121°C. Aseptically add amino acid and vitamin solutions. Mix thoroughly. Aseptically distribute 7.0mL amounts into tubes that contain meat particles (1 part meat particles to 5 parts fluid). Use: For the cultivation of Treponema brennaborense. Chopped Meat Medium with 0.025% Tween™ 80 (ATCC Medium 1228) Composition per 1230.0mL: Peptone 30.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl·H 2 O 0.5g Tween™ 80 0.25g Chopped meat extract filtrate 1.0L Chopped meat extract solids 200.0mL Resazurin (0.025% solution) 4.0mL pH 7.0 ± 0.2 at 25°C Chopped Meat Extract: Composition per liter: Beef or horse meat 500.0g NaOH (1N solution) 25.0mL Preparation of Chopped Meat Extract: Use lean beef or horse meat. Remove fat and connective tissue. Grind. Add meat and NaOH to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling while stirring. Cool to 25°C. Remove fat from surface. Filter. Reserve ground meat particles and filtrate. Add distilled/deion- ized water to filtrate and bring volume to 1.0L. Preparation of Medium: To 1.0L of chopped meat extract filtrate, add the remaining components, except the L-cysteine·HCl·H 2 O and chopped meat solids. Mix thoroughly. Gently heat to boiling. Cool to room temperature. Add the L-cysteine·HCl·H 2 O. Adjust pH to 7.0. Dis- © 2010 by Taylor and Francis Group, LLC 374 Chopped Meat Medium with 1% Tween™ 80 tribute 1 part chopped meat solids (by volume) and 5 parts of liquid (by volume) into tubes under O 2 -free 97% N 2 + 3% H 2 . Cap with rubber stoppers and place tubes in a press. Autoclave for 15 min at 15 psi pres- sure–121°C with fast exhaust. Use: For the cultivation and maintenance of Eubacterium and Lacto- bacillus species. Chopped Meat Medium with 1% Tween™ 80 (ATCC Medium 737) Composition per 1230.0mL: Peptone 30.0g Tween™ 80 10.0g K 2 HPO 4 5.0g Yeast extract 5.0g L-Cysteine·HCl·H 2 O 0.5g Chopped meat extract filtrate 1.0L Chopped meat extract solids 200.0mL Resazurin (0.025% solution) 4.0mL pH 7.0 ± 0.2 at 25°C Chopped Meat Extract: Composition per liter: Beef or horse meat 500.0g NaOH (1N solution) 25.0mL Preparation of Chopped Meat Extract: Use lean beef or horse meat. Remove fat and connective tissue. Grind. Add meat and NaOH to distilled/deionized water and bring volume to 1.0L. Gently heat and bring to boiling while stirring. Cool to 25°C. Remove fat from surface. Filter. Reserve ground meat particles and filtrate. Add distilled/deion- ized water to filtrate and bring volume to 1.0L. Preparation of Medium: To 1.0L of chopped meat extract filtrate, add the remaining components, except the L-cysteine·HCl·H 2 O and chopped meat solids. Mix thoroughly. Gently heat to boiling. Cool to room temperature. Add the L-cysteine·HCl·H 2 O. Adjust pH to 7.0. Dis- tribute 1 part chopped meat solids (by volume) and 5 parts of liquid (by volume) into tubes under O 2 -free 97% N 2 + 3% H 2 . Cap with rubber stoppers and place tubes in a press. Autoclave for 15 min at 15 psi pres- sure–121°C with fast exhaust. Use: For the cultivation and maintenance of Eubacterium biforme and Lactobacillus species. Christensen Agar Composition per liter: Agar 15.0g NaCl 5.0g Sodium citrate 3.0g KH 2 PO 4 1.0g L-Cysteine·HCl·H 2 O 0.1g Phenol Red 12.0mg pH 6.9 ± 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. Dispense into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Allow tubes to cool in a slanted position. Use: For the differentiation of enteric pathogens, especially members of the Enterobacteriaceae, and coliforms based on their ability to utilize citrate as a carbon source. Bacteria that can utilize citrate turn the medium pink-red. Christensen Agar Composition per liter: Agar 15.0g NaCl 5.0g Sodium citrate 3.0g KH 2 PO 4 1.0g Yeast extract 0.5g Glucose 0.2g L-Cysteine·HCl·H 2 O 0.1g Phenol Red 12.0mg pH 6.9 ± 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. Dispense into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Allow tubes to cool in a slanted position. Use: For the differentiation of enteric pathogens, especially members of the Enterobacteriaceae, and coliforms based on their ability to utilize citrate as a carbon source. Bacteria that can utilize citrate turn the medium pink-red. Christensen Citrate Agar (BAM M39) Composition per liter: Agar 15.0g NaCl 5.0g Sodium citrate 3.0g KH 2 PO 4 1.0g Yeast extract 0.5g Ferric ammonium citrate 0.4g L-Cysteine·HCl·H 2 O 0.1g Na 2 S 2 O 5 0.08g Phenol Red 12.0mg pH 6.9 ± 0.2 at 25°C Preparation of Medium: Add components to distilled/deionized wa- ter and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Dispense into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. Allow tubes to cool in a slanted position. Use: For the differentiation of enteric pathogens, especially members of the Enterobacteriaceae, and coliforms based on their ability to utilize citrate as a carbon source. Bacteria that can utilize citrate turn the medium pink-red. Christensen Citrate Agar, Modified (Citrate Agar) Composition per liter: Agar 12.0g NaCl 5.0g Sodium citrate 3.8g KH 2 PO 4 1.0g Yeast extract 0.5g Glucose 0.2g L-Cysteine·HCl·H 2 O 0.1g Phenol Red 0.02g pH 6.7 ± 0.2 at 25°C © 2010 by Taylor and Francis Group, LLC . 7.0mL into tubes that contain meat particles (1 part meat particles to 5 parts fluid). Autoclave for 30 min at 15 psi pressure– 121°C. Use: For the cultivation of various anaerobes. Chopped Meat. 7.0mL into tubes that contain meat particles (1 part meat parti- cles to 5 parts fluid). Autoclave for 30 min at 15 psi pressure–121°C. Use: For the cultivation of various anaerobes. Chopped Meat. 7.0mL into tubes that contain meat particles (1 part meat particles to 5 parts fluid). Autoclave for 30 min at 15 psi pressure–121°C. Use: For the cultivation of numerous anaerobes, including Actinomy- ces