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Andersen’s Pork Pea Agar

[所属分类:培养基配方] [发布时间:2021-11-1] [发布人:网站管理员2] [阅读次数:] [返回]

山东拓普生物工程有限公司培养基配方 http://www.topbiol.com

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/deionized 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 distilled/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 diatoma

ceous 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 com

ponents to distilled/deionized water and bring vol

ume to 1.0L. Mix thoroughly. Gently heat and bring

to boiling. Autoclave for 15 min at 15 psi pressure–

121°C. Cool to 45°–50°C.

NaHCO3 Solution:

Composition per 100.0mL:

NaHCO3 ..............................................................5.0g

Preparation of NaHCO3 Solution: Add

NaHCO3 to distilled/deionized water and bring vol

ume 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. Autoclave 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 suction. Autoclave for 12 min at 15 psi

pressure–121°C. Cool to 45°–50°C. Aseptically add

25.0mL of sterile NaHCO3 solution. Mix thoroughly.

Pour into sterile Petri dishes in 15.0mL volumes. Al

low agar to solidify. Cover agar with 10.0mL of ster

ile, cooled thioglycolate agar.

Use: For the cultivation of mesophilic Clostridium

species. For the recovery of endospores from foods

following heat treatments.

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Andersen’s Pork Pea Agar