微生物培養(yǎng)基配制完成后為什么必須立即滅菌處理？
發(fā)布時間：
2022-12-24
作者：
國內(nèi)微生物國產(chǎn)培養(yǎng)基品牌
答：這要從產(chǎn)品本身說起，因為培養(yǎng)基是做微生物細菌檢測用的，通俗講，就是在藥品，食品，水，化妝品等生產(chǎn)環(huán)境是否符合國家標準，是否符合生產(chǎn)環(huán)境要求，需要各種配置的培養(yǎng)基來檢測，是給細菌的營養(yǎng)品，供微生物生長、繁殖、代謝的混合養(yǎng)料。
如果成品培養(yǎng)基制作以后沒有及時滅菌，會造成污染細菌感染，降低產(chǎn)品使用率，在檢測的時候直接不合格不達標準，嚴重的時候培養(yǎng)基直接死亡。
由于微生物具有不同的營養(yǎng)類型，對營養(yǎng)物質(zhì)的要求也各不相同，加之實驗和研究的目的不同，所以培養(yǎng)基的種類很多，使用的原料也各有差異。為了達到實驗和研究的目的，要按照培養(yǎng)基的配方進行配制。一般培養(yǎng)基的滅菌采用高壓蒸汽滅菌、過濾滅菌。
1、高壓蒸汽滅菌：
高壓蒸汽滅菌適合于耐高溫培養(yǎng)基、接種器械和蒸餾水的滅菌。培養(yǎng)基在愛制備過程中混入各種雜菌，分裝后應(yīng)立即滅菌，至少應(yīng)在24h內(nèi)完成滅菌工作。滅菌時一般是在0.105MPa壓力下，溫度121℃時，滅菌15~30min即可。消毒時間不宜過長，也不能超過規(guī)定的壓力范圍，否則有機物質(zhì)特別是維生素類物質(zhì)就會在高溫下分解，失去營養(yǎng)作用，也會使培養(yǎng)基變質(zhì)、變色，甚至難以凝固。將蒸餾水或自來水裝在三角瓶中，裝水量一般不超過瓶的2/3，用牛皮紙或硫酸紙包扎封口，然后放入高壓鍋中滅菌后即為無菌水。 滅菌后的培養(yǎng)基可放到培養(yǎng)室中預(yù)培養(yǎng)3d，若無污染現(xiàn)象，則證明滅菌是徹底的，可以使用。暫時不用的培養(yǎng)基最好置于10℃下保存。含IAA或GA3的培養(yǎng)基應(yīng)在1周內(nèi)用完，其他培養(yǎng)基最多也不要超過1個月。
2、過濾滅菌：
一些植物生長調(diào)節(jié)劑及有機物，如IAA、GA3、ZT、CM等，遇熱容易分解，不能與培養(yǎng)基一起進行高溫滅菌，而要使用細菌過濾器濾去其中的雜菌。細菌過濾器與濾膜（孔徑0.45微米）使用之前要先進行高壓滅菌。過濾后的溶液要立即加入培養(yǎng)基中，若為液體培養(yǎng)基，可在培養(yǎng)基冷卻至30℃時加入；若為固體培養(yǎng)基，必須在培養(yǎng)基凝固之前（50-60℃）加入，振蕩使溶液與其他成分混合均勻。
Why must the microbiological medium be sterilized immediately after the preparation is completed?
Answer: this should be said from the product itself, because the culture medium is used for microbiological bacteria detection, popularly speaking, is the production environment such as medicine, food, water, cosmetics and other production environment conform to the national standard, whether it meets the requirements of the production environment, and needs various collocated culture medium to test the bacteria and provide the microorganism for the bacteria. A mixture of growth, reproduction and metabolism.
If the product culture medium is not sterilized in time, it will cause the infection of contaminated bacteria, reduce the use rate of the product, the direct disqualification at the time of testing, and the direct death of the culture medium at the time of serious.
As microbes have different types of nutrition, the requirements for nutrients are different, and the purpose of the experiment and research is different, so there are many kinds of culture medium and different materials used. In order to achieve the purpose of experiment and research, we should prepare according to the formula of the medium. Sterilization of general medium is carried out by high pressure steam sterilization and filtration sterilization.
1. Autoclave sterilization:
High pressure steam sterilization is suitable for sterilization of high temperature medium, inoculating apparatus and distilled water. The medium is mixed with all kinds of heterozygous bacteria in the process of preparation, and should be sterilized immediately after being packed. At least the sterilization should be done in 24h. Sterilization is usually done under 0.105MPa pressure and at 121 C, 15~30min can be sterilized. The time of disinfection should not be too long or more than the prescribed pressure range. Otherwise, the organic matter, especially the vitamins, will decompose under high temperature, lose the nutrition, and make the medium metamorphic, discoloration, and even hard to solidify. The distilled water or tap water is installed in a triangle bottle. The amount of water is generally not more than 2/3 of the bottle. It is sealed with kraft paper or sulphuric acid paper, and then put into the high pressure pot and sterilized, that is, aseptic water. After sterilization, the culture medium can be pre incubated in the culture room 3D. If there is no pollution, it is proved that sterilization is thorough and can be used. The culture medium which is temporarily not used is best kept at 10 degrees Celsius. Medium containing IAA or GA3 should be used in 1 weeks, and other culture media should not exceed 1 months at most.
2, filter sterilization:
Some plant growth regulators and organic compounds, such as IAA, GA3, ZT, CM and so on, are easily decomposed with heat, and can not be sterilized with the culture medium at high temperature, but bacteria filters should be used to filter out the bacteria. Bacterial filter and filter membrane (aperture 0.45 micron) should be autoclaved before being used. The filtrated solution should be added into the medium immediately. If it is a liquid medium, it can be added to the medium when the medium is cooled to 30. If it is a solid medium, it must be added before the medium solidification (50-60 degrees C), and the oscillation can make the solution mixed evenly with the other components.