Development Of Probiotics Powder And The Influence Of Compound Probiotics Powder On The Growth Performance In Mice

Probiotics, which has benefical effect on host through improving the balance ofmicroorganism colonies, could produce funtional materials. Probiotics is a most importantbacteria in intestinal canal. Probiotics has mutiple functions such as adjust the balance ofmicroorganisms in intestinasl canal, enhance immunity and produce biological activitymetabolite.In this thesis, Bacillus subtilis natto, Lactobacillus acidophilus and Bifidobacteriumwere used as wild strains to start the following experiments:(1) The culture conditions of Bacillus subtilis natto were optimized, and the powderforms for optimally attaining the beneficial effects of Bacillus subtilis natto were prepared,which facilitates the further development of Bacillus subtilis natto, into apotentially usefulhealth-promoting probiotic product with a novel antithrombotic activity. The optimumcomposition of protectant for Bacillus subtilis natto was Glycerin2%, skimmed milkpowder15%, gelatin2%, trehalose12%. The optimum survival rate of the powder formscould be maintained at the level of up to77%, which enables the potential probioticproduct to produce its activities at sufficiently protection levels.(2) The compositions of vacuum freeze-drying protectant for cell growth ofLactobacillus acidophilus and Bifidobacterium were optimized using "one factor at a time"test and L9(34) orthogonal experiment. Results indicated that Lactobacillus acidophilusstayed in adaptive phase in0-8h, log phase in8-24h, stable phase in24-40h and thendecline phase. Bifidobacterium was in adaptive phase in0-8h, log phase in8-20h, stablephase in20-40h and then decline phase. The optimum composition of protectant forlactobacillus acidophilus was trehalose12%, skimmed milk powder5%, sucrose5%,gelatin2%. The optimum composition of protectant for Bifidobacterium was gelatin1.5%,skimmed milk powder15%, trehalose10%, sucrose5%.(3) The properties such as viable count, crude protein content, total sugar contents andacidity values of Bacillus subtilis natto, Lactobacillus acidophilus and Bifidobacterium infreeze-drying state were determined. The tolerance of these strains in freeze-drying state inaritificial simulate succus gastricus, succus entericus and choler and overall stomach andintestine environment in simulation was also determined. As results shown, the parametersfor Bacillus subtilis natto were viable count10.9×109cfu/g, moisture content2.99%, totalsugar35.69%, crude protein18.31%and acidity0.36%, parameters for Lactobacillusacidophilus were viable count9.26×109cfu/g, moisture content3.09%, total sugar42.16%,crude protein5.17%and acidity0.83%, parameters for Bifidobacterium were viable count8.48×109cfu/g, moisture content2.09%, total sugar44.08%, crude protein16.05%andacidity0.87%. After45min of soaking in aritificial simulate succus entericus, survival rates of Bacillus subtilis natto, Lactobacillus acidophilus and Bifidobacterium were88.66%,89.53%,84.68%, respectively. After30min of soaking in aritificial simulatecholer, survival rates of Bacillus subtilis natto, Lactobacillus acidophilus andBifidobacterium were86.85%,87.72%and84.70%, respectively. After30min of soakingin aritificial simulate overall stomach and intestine environment, survival rates of Bacillussubtilis natto, Lactobacillus acidophilus and Bifidobacterium were84.45%,84.37%and83.15%, respectively. After experiments, the results suggested Bacillus subtilis natto,Lactobacillus acidophilus and Bifidobacterium in freeze-drying state should be stored at4℃.(4) Mice were feeding by those probiotics powders with1:1:1of Bacillus subtilis natto,Lactobacillus acidophilus and Bifidobacterium in freeze-drying state for30d. The growthparameters lymphoid cells, neutral particles, serum and blood routine index were tested tofind out the effects of compound probiotics powder on growth of mice. The resultsindicated that the increasing weight and lymphoid cells, neutral particles of control setwere lower than those of all three experimental groups feeding with compound probioticspowder. The three experimental groups setting low, medium and high dose have a largerincreasing weight with higher food intake. The results showed that the compoundprobiotics powder has ability to promote the appetite of mice. The wet weight of liver,kidney, spleen and organ index of medium and high dose groups were higher than those oflow dose one. That indicated that compound probiotics powder could promote the growthof liver, kidney and spleen.