Study On The Viable But Nonculturable State Bifidobacterium Adolescentis

The Bifidobacterium adolescentis is probiotics,mainly be applied in the new green feed additive which appeared in the recent years.It has the characteristic of the non-toxic side effect,non-residual, non-drug resistance,non-pollute-environment and so on.Its main function is supplementing microorganism in the quantity either the type which the intestinal tract reduced,to be restore microbiology balance of the intestinal tract,achieve the goal of health care."The viable but nonculturable state"is a kind of special survival state when the bacterium under the poor surrounding conditions.The bacterium could not be cultured by normal medium,but it is still viable and has metabolic activity.Under the adequate condition,the bacterium could recover the culturable state.The purpose of the experiment is researching viable but non-culturable state Bifidobacterium adolescentis,take it into the research area of probiotics, find a kind of method in producing new feed additive which prevents illness.We did research on the state of viable but non-culturable to the Bifidobacterium adolescentis bought from China Center of Industrial Culture Collection.With many kinds of detection ways.Finally we got the stable state that Bifidobacterium adolescentis can be induced to viable but non-culturable state.And we did psychology-biochemistry and 16sRNA test and so on..We proved the bacterium used to be induced was Bifidobacterium adolescentis.Bifidobacterium adolescentis in the liquid LB culture medium,pH=7,puts in 4℃under low temperature anaerobic static cultivation then enters Viable but nonculturable state.The 15 days later Bifidobacterium adolescentis enters Viable but nonculturable state,The state can maintains as long as100 days.The LB culture medium carries on the induction compared with the medium of TPY,it can save the resources,reduces the cost,and compounds simply,easy to operate,easy to preserve.Using the examination of physiological biochemistry character and the appraisal is the most early method for detection and appreciation Bifidobacterium adolescentis,its also the credible method at present.The result of appreciation is coincidence that the target which responded for all physiological biochemistry tallies completely with the Bifidobacterium adolescentis theory target.It may determine this bacterium initially for Bifidobacterium adolescentis.But bacterium's 16S rRNA constant region sequence is basic conservative,therefore using Bifidobacterium adolescentis constant region sequence to design primer and to amplificate the 16SrRNA part,and may through this sequence to analysis,its homology to be 99.2%, the last determines the bacterium which finally uses in experimenting as Bifidobacterium adolescentis.