Andersen’s Pork Pea Agar

[所属分类:培养基配方] [发布时间:2019-10-17] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Andersen’s Pork Pea Agar
Composition per 1685.0mL:
Agar ............................................................................................16.0g
Peptone..........................................................................................5.0g
Pancreatic digest of casein............................................................1.6g
K2HPO4 .......................................................................................1.25g
Soluble starch................................................................................1.0g
Sodium thioglycolate....................................................................0.5g
Pork infusion..........................................................................800.0mL
Thioglycolate agar .................................................................660.0mL
Pea infusion............................................................................200.0mL
NaHCO3 solution.....................................................................25.0mL
pH 7.2 ± 0.2 at 25°C
Pork Infusion:
Composition per liter:
Pork, fresh lean ground.............................................................454.0g
Preparation of Pork Infusion: Add ground pork to distilled/de-
ionized water and bring volume to 1.0L. Autoclave for 60 min at 0 psi
pressure–100°C. Filter through two layers of cheesecloth. Cool to 4°C.
Skim fat from surface. Warm to 25°C. Centrifuge at 5000 rpm for 10
min. Discard pellet.
Pea Infusion:
Composition per 450.0mL:
Green peas, fresh or frozen.......................................................454.0g
Diatomaceous earth (celite)........................................................10.0g
Preparation of Pea Infusion: Add green peas to 450.0mL of dis-
tilled/deionized water. Blend until smooth. Autoclave for 60 min at 0
psi pressure–100°C. Centrifuge at 5000 rpm for 10 min. Discard pellet.
Clarify supernatant solution with diatomaceous earth (celite). Filter
through Whatman #4 filter paper. Use filtrate solution.
Thioglycolate Agar:
Composition per liter:
Agar ..........................................................................................20.75g
Pancreatic digest of casein..........................................................15.0g
Glucose .........................................................................................5.5g
Yeast extract..................................................................................5.0g
NaCl..............................................................................................2.5g
L- Cystine .......................................................................................0.5g
Sodium thioglycolate....................................................................0.5g
Resazurin ...................................................................................1.0mg
pH 7.1 ± 0.2 at 25°C
Preparation of Thioglycolate Agar: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gen-
tly heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 45°–50°C.
NaHCO 3 Solution:
Composition per 100.0mL:
NaHCO3 ........................................................................................5.0g
Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize.
Preparation of Medium: Combine components, except NaHCO3
solution and thioglycolate agar. Mix thoroughly. Adjust pH to 7.2. Au-
toclave for 5 min at 15 psi pressure–121°C. While medium is still hot,
add 25.0g of celite. Filter through Whatman #4 filter paper with suc-
tion. Autoclave for 12 min at 15 psi pressure–121°C. Cool to 45°–
50°C. Aseptically add 25.0mL of sterile NaHCO 3 solution. Mix thor-
oughly. Pour into sterile Petri dishes in 15.0mL volumes. Allow agar to
solidify. Cover agar with 10.0mL of sterile, cooled thioglycolate agar.
Use: For the cultivation of mesophilic Clostridium species. For the
recovery of endospores from foods following heat treatments.