Development Of Foxtail-like Sophara Seed’s Dregs-Bifidobacterium Bifidum-lactobacillus Salivarius Synbiotics Microcapsule

In order to develop a safe, high effective and green type synbiotics microcapsule, which can be used for preventing Early Weanling Stress Syndrome of Piglets. Lactobacillus salivarius and Bifidobacterium bifidum, which can both inhibit piglet pathogenic Escherichia coli, were used as probiotics. While, foxtail-like sophara seed’s dregs was used as prebiotics. In this study, we discussed the fabrication process of synbiotics microcapsule produced by alginate-chitosan-coated methods.The test follows the steps below:1. The isolation of advantage lactic acid bacteria, which can inhibit piglet pathogenic Escherichia coli. We collected different-day old healthy piglets’ fresh feces samples, after separating, purifying and Oxford cup method screening. At last, two strains were screened out.2. Studies on the characteristics of the screening strains. We identified the screening strains by means of biochemical identification and16S rDNA homology analysis. Then we researched their biological characteristics. Finally, it was compared with a Lactobacillus acidophilus standard strain separated from pig rectum, which has strong bacteriostatic effect on pathogenic Escherichia coli reported by literatures.3. The formulation of synbiotics microcapsule. On the base of a large amount of tests and abundant comparisons, we found out the proportion of probiotics, prebiotics and the concentration of protective agent, calcium chloride and shell materials4. The preparation of synbiotics microcapsule. We adopted three differents methods to prepare foxtail-like sophara seed’s dregs-Bifidobacterium bifidum-lactobacillus salivarius synbiotics microcapsule, and then we did studies on their characteristics, such as storage stability.The results showed that:1. We separated35suspected lactic acid bacteria strains from healthy piglets’fresh feces samples using TPY and MRS medium under anaerobic conditions. Finally,11#and14#strains, which have better antibacterial effect on piglet pathogenic Escherichia coli, were screened out by Oxford cup method.2.11#and14#strains were all identified as Lactobacillus salivarius by means of biochemical identification and molecular identification. Considering the results of Oxford cup method test and the studies on their biological characteristics,14#strain was used as probiotics for synbiotics microcapsule. Moreover,14#strain had far greater bacteriostatic effect than Lactobacillus acidophilus standard strain separated from pig rectum, which has strong bacteriostatic effect on pathogenic Escherichia coli, reported by literatures.3. Compared with several common microcapsule fabrication techniques, the extrusion method was suitable for coating Bifidobacterium bifidum and lactobacillus salivarius. The optimum condition for synbiotics microcapsule was established as follows:probiotics (The living bacterium ratio of Bifidobacterium bifidum and Lactobacillus salivarius is10:1) adhesive (4%skim milk,1%L-glutamate,2%Sodium alginate,2.5%CaCl2,0.5%chitosan and6%foxtail-like sophara seed’s dregs)=2:1。4. The viable bacteria counts of method1and method3can reached1.1×109Cfu/g and2.0×108Cfu/g respectively. However, when they were orderly stayed in artificial gastric juice and artificial intestinal fluid for2hours, the viable bacteria counts reached2.5×108Cfu/g and4.3×107Cfu/g respectively. The results showed that the synbiotics microcapsule prepared by method1and method3had high viable bacteria counts, long shelf life and they were convenient for storage at room temperature. What’s more, they also had acid resistance, enteric properties and no gastric solubility. Conclusion:1.11#and14#strains separated from healthy piglets’fresh feces samples, which have best antibacterial effect on pathogenic Escherichia coli were all identified as Lactobacillus salivarius.2. The optimum conditions for synbiotics microcapsule prepared by the extrusion method were established as follows:probiotics (The living bacterium ratio of Bifidobacterium bifidum and Lactobacillus salivarius is10:1):adhesive (4%skim milk,1%L-glutamate,2%Sodium alginate,2.5%CaCl2,0.5%chitosan and6%foxtail-like sophara seed’s dregs)=2:1.3. Synbiotics microcapsule prepared by method1and method3have the advantages of being simple, steady performance, convenient for industrialization produce, transporting, storage, clinical application and other aspects.