Actinobacillus lignieresii Medium

[所属分类:培养基配方] [发布时间:2019-9-23] [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

Actinobacillus lignieresii Medium
Composition per 1010.0mL:
Agar............................................................................................10.0g
Hartley’s digest broth.............................................................900.0mL
Filde’s enrichment .................................................................100.0mL
Antibiotic solution...................................................................10.0mL
pH 7.5 ± 0.2 at 25°C
Hartley’s Digest Broth:
Composition per 10.0L:
Ox heart ..................................................................................3000.0g
Pancreatin ...................................................................................50.0g
Na 2 CO 3 , anhydrous (0.8% solution).............................................5.0L
HCl, concentrated....................................................................80.0mL
Preparation of Hartley’s Digest Broth: Finely mince the ox
heart. Add the meat to 5.0L of distilled/deionized water. Gently heat
and bring to 80°C. Add Na 2 CO 3 solution. Cool to 45°C. Add pancre-
atin and maintain at 45°C for 4 hr while stirring. Add the HCl and
steam at 100°C for 30 min. Cool to room temperature. Adjust pH to 8.0
with 1N NaOH. Gently heat and bring to boiling. Continue boiling for
25 min. Filter while hot through Whatman #1 filter paper. Cool to room
temperature. Adjust pH to 7.5.
Filde’s Enrichment Solution:
Composition per 206.0mL:
Pepsin............................................................................................1.0g
NaCl (0.85% solution)...........................................................150.0mL
Sheep blood, defibrinated........................................................50.0mL
HCl.............................................................................................6.0mL
Source:  Filde’s enrichment solution is available as a premixed powder
from BD Diagnostic Systems and Oxoid Unipath.
Preparation of Filde’s Enrichment Solution: Combine compo-
nents. Mix thoroughly. Incubate at 56°C for 4 hr. Bring pH to 7.0 with
20% NaOH. Adjust pH to 7.2 with HCl. Do not autoclave. Add 0.25
mL of chloroform and store at 4°C. Before use, heat to 56°C to remove
chloroform.
Antibiotic Solution:
Composition per 10.0mL:
Oleandomycin phosphate............................................................0.02g
Neomycin sulfate.......................................................................1.5mg
Preparation of Antibiotic Solution: Add components to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Filter
sterilize.
Preparation of Medium: Add agar to 900.0mL of Hartley’s digest
broth. 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
100.0mL of Filde’s enrichment solution and 10.0mL of antibiotic solu-
tion. Mix thoroughly. Pour into sterile Petri dishes or distribute into
sterile tubes.
Use: For the isolation and cultivation of Actinobacillus lignieresii.