改良亮绿琼脂

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规格: 250g 500g

联系方式:I47-825O-882O

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改良BG琼脂产品基本信息

产品名称: 改良亮绿琼脂;含磷酸盐的亮绿琼脂;根据SLMB方法的亮绿琼脂;BPLS琼脂
英文名称: Brilliant Green Agar, Modified;Brilliant Green Agar with Phosphates;Brilliant Green Agar acc. SLMB;BPLS Agar
培养基类型: 选择与鉴别培养基
级别: for microbiology
品牌: ELITE-MEDIA
产品目录号: M710-01、M710-02
产品规格: 250g、500g 添加剂需另购
产品外观: 米色至绿色均一粉末。
颜色与澄清度: 绿色至棕色凝胶。
保存条件: 密封,2-25°C保存。
注意事项: 避免摄入、呼入和皮肤接触。
相关产品: --


产品描述:

改良亮绿琼脂(Brilliant Green Agar, modified)即根据SLMB方法的亮绿琼脂(Brilliant Green Agar acc. SLMB),含有磷酸盐,是一种高度选择性培养基,用于分离和鉴定水、污水和食品中的非伤寒沙门氏菌。亮绿琼脂不能分离伤寒杆菌和志贺氏菌。
这个配方抑制剂浓度增加,选择性增强,被国际标准组织(ISO)、欧洲共同体标准方法、美国公众健康协会(APHA) 、官方分析化学师协会(AOAC)引用推荐用于分离和鉴别沙门氏菌。
改良煌绿琼脂相对于其它配方的优势在于对大肠埃希氏菌和变形杆菌的抑制作用更强;限制菌落形态与沙门氏菌相似的假单胞菌的生长,降低假阳性率;对数量少的沙门氏菌没有抑制性。



用途:

改良亮绿琼脂用于从污水、污泥、海鸥粪便中分离沙门氏菌。
改良亮绿琼脂用于检测食品和饲料中的沙门氏菌。
改良亮绿琼脂用于临床诊断,分离和鉴定粪便等材料中的沙门氏菌。



原理:

亮绿琼脂(Brilliant Green Agar)也称煌绿琼脂、亮绿酚红乳糖蔗糖琼脂,简称BG琼脂,是一种强选择性培养基,用于从病理材料、食品、奶制品中分离和鉴定沙门氏菌属(伤寒杆菌除外)。亮绿琼脂是Kristensen等人最早研制的,作为沙门氏菌选择性分离培养基。

亮绿琼脂也是一种基于肠道菌对乳糖-蔗糖发酵能力进行鉴别的培养基。亮绿染料是抑制剂,抑制革兰氏阳性菌和大多数革兰氏阴性杆菌生长,对绝大多数沙门氏菌无抑制作用。酚红作为酸碱指示剂,酸性条件下显黄色,碱性条件下显红色。乳糖-蔗糖发酵产酸的细菌,菌落显黄绿色,四周培养基显黄绿色。沙门氏菌和其它非乳糖-蔗糖发酵型微生物菌落呈红色、粉色或白色,四周培养基呈亮红色。



配方与配制方法:

成分 g/L
肉浸粉 5.0
细菌学蛋白胨 10.0
酵母提取物 3.0
磷酸氢二钠 1.0
磷酸二氢钠 0.6
乳糖 10.0
蔗糖 10.0
酚红 0.09
亮绿 0.0047
琼脂 12.0
Final pH 6.9 +/- 0.2  

配制方法:
称取52g煌绿琼脂,加1000 mL超纯水,在火焰上方加热煮沸完全溶解。不要高温蒸汽灭菌!
冷却至50度左右时,倒平板。待干燥后使用



实验方法:

亮绿琼脂培养基应该与初步筛选培养基(如麦康凯琼脂)和富集增菌用培养基(如四磺酸盐肉汤)协同使用。

未经处理的粪便样本放置3h后,沙门氏菌的检出率将严重降低。来不及检测的粪便样品接种到转移培养基上,以维持微生物的活性。

向亮绿琼脂中添加磺胺类药物(0.8g或1g)能够提高沙门氏菌检出率。加抗生素的改良煌绿琼脂对污染微生物的抑制作用更强,假阳性的概率更低。用于从污水、污泥、海鸥粪便中分离沙门氏菌。已报道的抗生素使用组合有乙酰磺胺(1.0 mg/ml)和扁桃酸(0.25 mg/ml) ,磺胺乙酰钠(1.0g/L)和扁桃酸钠(0.35 g/L)。去氧胆酸钠(2.5 g/L) 可以阻止变形杆菌的疯长。


Organisms (ATCC) 生长情况 菌落颜色
Salmonella thiphimurium (14028) +++ 红色至白粉色
Salmonella enteritidis (13076) +++ 红色至白粉色
Salmonella typhi (6539) + 红色至粉色
Staphylococcus aureus (25923) - -
Escherichia coli (25922) -或部分 绿色

食品和饲料中沙门氏菌的检测方法

1. One part of the food sample was added to 20 parts of Muller-Kauffmann Tetrathionate Medium.
2. After agitation, the flask of broth was placed into a 45°C waterbath for 15 minutes only.
3. The flask was then transferred to a 43°C incubator.
4. The broth was subcultured to Brilliant Green Agar (Modified) after 18 and 48 hours.
A single loopful of broth was used to streak inoculate either two 9 cm diameter plates (without recharging the loop between plates) or one 14cm diameter plate.
5. The plates were incubated at 35°C for 18-24 hours.
6. Red colonies, resembling salmonellae, were picked off the plates and subcultured to Lysine Decarboxylase Broth and Triple Sugar Iron Agar. These media were incubated at 35°C for 18-24 hours.

If the reactions on these media were positive for salmonellae then slide agglutination tests were carried out on the surface growth of the Triple Sugar Iron Agar.

污水中沙门氏菌的检测方法

1. Take a representative sample of sewage or sludge for examination.
2. Homogenise a suitable volume in a macerator or stomacher.
3. Inoculate five 10ml samples into 35ml of Buffered Peptone Water, five 1ml samples and five 0.1ml samples into 10ml of Buffered Peptone Water. Incubate at 35°C overnight.
4. Transfer 10ml portions into 35ml of Muller-Kauffmann Tetrathionate Broth and incubate at 43°C.
5. Subculture the broths on to Brilliant Green Agar (Modified) containing Sulphamandelate Selective Supplement after 24 and 48 hours incubation.
6. Incubate the Brilliant Green Agar plates overnight at 43°C.
7. Identify suspicious (red) colonies using further diagnostic tests.

The Sulphamandelate Selective Supplement inhibits competing organisms which multiply during the resuscitation and recovery stages in Buffered Peptone Water.


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