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生化培養箱在多管發(fā)酵法

更新時(shí)間:2011-11-10      點(diǎn)擊次數:963

2.濾膜法
濾膜是一種微孔性濾膜,孔徑0.45μm。將水樣注入已滅菌的放有濾膜的過(guò)濾器中,經(jīng)過(guò)抽濾,細菌即被截留在膜上,然后將濾膜貼于M-FC培養基上,在44.5℃±0.5℃下培養24 h±2h。糞大腸菌群菌落在M-FC培養基上呈藍色或藍綠色,其他非糞大腸菌群菌落成灰色、淡黃色或無(wú)色。正常情況下,由于溫度和玫瑰酸鹽試劑的選擇性作用,在M-FC培養基上很少見(jiàn)到非糞大腸菌菌落。

1.多管發(fā)酵法

初發(fā)酵實(shí)驗  發(fā)酵管內裝有單倍或三倍乳糖蛋白胨培養液,并在內倒置有小玻璃管。每個(gè)樣品用三個(gè)不同的水樣量接種,同一接種水樣量要有五管,將水樣分別接種到裝有乳糖蛋白胨培養液的發(fā)酵管中,在37℃±0.5℃下培養24h±2h。乳糖能起選擇作用,因為很多細菌不能發(fā)酵乳糖,而大腸菌群能發(fā)酵乳糖并產(chǎn)酸產(chǎn)氣。培養液中加入溴甲酚紫作為酸度指示劑,細菌產(chǎn)酸后,培養液即由原來(lái)的紫色變成黃色。產(chǎn)酸產(chǎn)氣的發(fā)酵管表明試驗陽(yáng)性,如在倒管內產(chǎn)氣不明顯,可輕拍發(fā)酵管,有小氣泡升起為陽(yáng)性。
復發(fā)酵實(shí)驗  輕微振蕩初發(fā)酵試驗陽(yáng)性結果的發(fā)酵管,用3mm接種環(huán)將培養物(2-3環(huán))轉接到EC培養液中,在44.5℃±0.5℃下培養24h±2h(提高培養溫度可造成不利于來(lái)自自然環(huán)境的大腸菌群的生長(cháng))。培養后立即觀(guān)察,發(fā)酵管產(chǎn)氣則證實(shí)為糞大腸菌群陽(yáng)性。

 

 

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