Oxoid CM0877B PALCAM琼脂基础 PALCAM AGAR BASE

Oxoid CM0877B PALCAM琼脂基础 PALCAM AGAR BASE

Dehydrated Culture Media

BUFFERED LISTERIA ENRICHMENT BROTH

Code: CM0897

A selective enrichment medium for the detection of Listeria monocytogenes when prepared from Buffered Listeria Enrichment Broth base and Listeria Selective Supplement SR0141.

由 Buffered Listeria Enrichment Broth base 和 Listeria Selective Supplement SR0141 制备的用于检测单核细胞增生李斯特菌的选择性富集培养基。

BUFFERED LISTERIA ENRICHMENT BROTH

Typical Formula*

gm/litre

Tryptone soya broth

30.0

Yeast extract

6.0

Potassium di-hydrogen orthophosphate

1.35

Disodium hydrogen orthophosphate

9.60

Final pH 7.3 ± 0.2 @ 25°C
* Adjusted as required to meet performance standards

LISTERIA SELECTIVE ENRICHMENT SUPPLEMENT

Code: SR0141

Vial contents: (each vial is sufficient for 500ml of medium)
per vial
per litre
Nalidixic acid

20.0mg

40.0mg
Cycloheximide

25.0mg

50.0mg
Acriflavine hydrochloride

7.5mg

15.0mg

MODIFIED LISTERIA SELECTIVE ENRICHMENT SUPPLEMENT

Code: SR0213

Vial contents: (each vial is sufficient for 500ml of medium)
per vial
per litre
Nalidixic acid

20.0mg

40.0mg
Amphotericin B

5.0mg

10.0mg
Acriflavine hydrochloride

7.5mg

15.0mg

Directions
Add 23.5g to 500ml of distilled water and mix well to dissolve. Add the contents of 1 vial of Listeria Selective Enrichment  Supplement SR0141 or the alternative Modified Listeria Selective Supplement SR0213 reconstitued as directed.  Sterilize by autoclaving at 121°C for 15 minutes. Cool to 50°C, mix well and aseptically distribute into sterile containers in volumes as required.

Description
Listeria Selective Enrichment Broth CM0862 is based on the formulation described by Lovett et al.1 and is recommended for the enrichment of Listeria species in food. Subsequent work has concluded that the enrichment properties can be improved by increasing the buffering capacity of the medium by the addition of potassium di-hydrogen orthophosphate and disodium hydrogen orthophosphate. Buffered Listeria Enrichment Broth is therefore a modification of the original medium.

Techniques
1. Add 25g or 25ml samples to 225ml of Buffered Listeria Enrichment Broth. Homogenise if required.
2. Incubate at 30°C for 48 hours.
3. Subculture from the Buffered Listeria Emrichment Broth onto Listeria Selective Agar plates (See Note) after 24 and 48 hours by:
(i) Direct plating onto Listeria Selective Agar plates.
(ii) Adding 1ml of the Buffered Listeria Enrichment Broth to 9ml of 0.5% KOH, vortex mixing, and plating onto Listeria Selective Agar plates.
Note
Suitable Listeria Selective Media are:
1. Listeria Selective Medium (Oxford formulation) (Oxoid CM0856 and Oxoid SR0140).
2. PALCAM Medium (Oxoid CM0877 and Oxoid SR0150)

Storage conditions and Shelf life
Buffered Listeria Enrichment Broth should be stored tightly capped in the original container in a cool, dry place away from bright light. When stored as directed the medium will remain stable until the expiry date printed on the label.

Listeria Selective Enrichment Supplement SR0141 as supplied should be stored at 2-8°C. When stored as directed the reagents are stable until the expiry date printed on the label.

Appearance
Dehydrated medium: Straw coloured, free-flowing powder
Prepared medium: Straw coloured solution

方向
将 23.5g 加入 500ml 蒸馏水中并充分混合溶解。添加 1 瓶李斯特菌选择性富集补充剂 SR0141 或替代的改良李斯特菌选择性补充剂 SR0213 的内容物,按指示重新配制。通过在 121°C 高压灭菌 15 分钟进行灭菌。冷却至 50°C,充分混合并按要求无菌分装到无菌容器中。

描述
李斯特菌选择性富集肉汤 CM0862 基于 Lovett 等人 1 描述的配方,推荐用于食品中李斯特菌的富集。随后的工作得出结论,通过添加正磷酸二氢钾和正磷酸氢二钠来增加培养基的缓冲能力,可以提高富集性能。因此,缓冲李斯特菌富集肉汤是对原始培养基的改良。

技术
1. 将 25g 或 25ml 样品加入 225ml 缓冲李斯特菌浓缩肉汤中。如果需要,均质化。
2. 30°C 孵育 48 小时。
3. 24 和 48 小时后,将缓冲的李斯特菌浓缩肉汤传代培养到李斯特菌选择性琼脂平板(见注)上:
(i) 直接铺板到李斯特菌选择性琼脂板上。
(ii) 将 1ml 缓冲李斯特菌富集肉汤加入 9ml 0.5% KOH,涡旋混合,然后铺板到李斯特菌选择性琼脂板上。
笔记
合适的李斯特菌选择性培养基是:
1. 李斯特菌选择性培养基(牛津配方)(Oxoid CM0856 和 Oxoid SR0140)。
2. PALCAM 培养基(Oxoid CM0877 和 Oxoid SR0150)

储存条件和保质期
缓冲李斯特菌浓缩肉汤应密封保存在原始容器中,置于阴凉干燥处,远离强光。当按照指示储存时,介质将保持稳定,直到标签上打印的有效期为止。

提供的李斯特菌选择性富集补充剂 SR0141 应在 2-8°C 下储存。当按指示储存时,试剂在标签上印刷的有效期之前是稳定的。

外貌
脱水介质:稻草色、自由流动的粉末
制备培养基:稻草色溶液

Quality control

Positive control: Expected results
Listeria monocytogenes ATCC® 19117 Turbid growth
Negative control:
Enterococcus faecalis ATCC® 29212* Inhibited
* This organism is available as a Culti-Loop®

Precautions
Listeria Selective Enrichment Supplement contains cycloheximide and is toxic if swallowed, inhaled or by skin contact. As a precaution when handling wear gloves and eye/face protection.
Acriflavine is activated by light and may form compounds inhibitory for Listeria.

References
1. Lovett J., Francis D.W. and Hunt J.M. (1987) J. Food Prot. 50. 188-192.
2. Curtis G.D.W., Nichols W.W. and Falla T.J. (1989) Lett. Appl. Micro. 8. 169-172.

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