Preparation And Application Of Novel Robiotics In Animal Husbandry

In this paper,the six probitics(Bacillus subtilis Lw-K16,Bacillus amyloliquefaciens Lw-J21,Auto chthonous flora Lw-R35,Bifidobacterium Lw-D48,Bacillus Subtilis Lw-X21,and Saccharomyces Lw-JM70)screened from ruminal sources of cattle and sheep were determined by streak plate method.The physiological characterizations of heat tolerance,acid resistant and bile salt tolerance of these probitics were evaluated,to determine whether the six probitics have the potential to become the sources of microecologics.And then,the six probitics were activated and cultured.The gastric irrigation was prepared and used to feed mice.The indicators involving mouse production performance,intestinal flora,fat accumulation,immune index,and blood glucose regulation were investigated and four optimal probitics(Auto chthonous flora Lw-R35,Bifidobacterium Lw-D48,Bacillus Subtilis Lw-X21,and Saccharomyces Lw-JM70)were screened from the six probitics in this paper.The four optimal probitics were immobilized using starch as a carrier and observed with electron microscopy as well as load measurement.The number of viable cells was measured over a certain period of time,and the result showed what the effect of the preservation time on the probiotics.The main conclusions were described as follows.(1)The colony morphology of six probitics was observed with microscope.It can be seen that there are large differences among them,and it is easy to distinguish.The antagonistic experiments showed that the antagonistic effect not obvious.Moreover,the growth curves of the 6 bacteria were described.(2)With determination of physiological characterizations,it can be found that Bacillussubtilis Lw-K16,Bacillus amyloliquefaciens Lw-J21 and Bacillus Subtilis Lw-X21exhibited good heat tolerance,while Auto chthonous flora Lw-R35,Bifidobacterium Lw-D48and Saccharomyces Lw-JM70 were poor.Therefore,consideing this point,the processing method such as freeze drying could be use.About acid resistance,Bacillus amyloliquefaciens Lw-J21,Auto chthonous flora Lw-R35,Bifidobacterium Lw-D48,Bacillus Subtilis Lw-X21,and Saccharomyces Lw-JM70 exhibited strong acid resistance.The survival rate of Bacillus subtilis Lw-K16 was only 26.30%after 4 h of storage at the value of pH is 2.0.At pH of 3.0,it showed strong acid resistance.Bacillus Subtilis Lw-X21,and Saccharomyces Lw-JM70were found to proliferate in a bile salt concentration of 0.3%.At the same time,Bacillus subtilis Lw-K16,Bacillus amyloliquefaciens Lw-J21,and Bifidobacterium Lw-D48 also showed some bile salt tolerance.The bile salt tolerance of Auto chthonous flora Lw-R35 was the worst in these six probiotics,and the content was only 53.7%after treatment for 12 h under the condition of 0.3%bile salt concentration.(3)All of the six probiotics can increase the weight of mice in different degrees,promote the utilization rate of mouse food for mice,and reduce the deposition of body fat.Moreover,there is no toxic side effect on the liver.With the experiment of the effect of mouse immune index,it was found that the effects of Bacillus amyloliquefaciens Lw-J21 and Bacillus Subtilis Lw-X21 on the spleen index and thymus index of mice were extremely significant(P<0.01).Auto chthonous flora Lw-R35 and Bacillus Subtilis Lw-X21 could significantly increase the number of intestinal beneficial strains in mice(P<0.05),improve the intestinal environment of mice,and maintain the balance of intestinal environment.After 12 hours of fasting,the mice fed with the probiotics were given glucose solution,and it was found that the mice fed with Bacillus amyloliquefaciens Lw-J21 had significantly higher blood glucose regulation(P<0.05)comparied with Bifidobacterium Lw-D48.(4)The four optimal probiotics were immobilized.With electron microscopy,it reveals that the probiotics were irregularly distributed on the surface of the carrier.Among them,Saccharomyces Lw-JM70 load was the smallest,which was approximately 3.81×10~8 CUF/g.After 7 weeks storage at room temperature,the viability of each group of bacteria was45.22%forBacillus amyloliquefaciens Lw-J21,53.85%for Auto chthonous flora Lw-R35,57.63%for Bacillus Subtilis Lw-X21,and 48.41%for Saccharomyces Lw-JM70,indicating that the immobilization can effectively prolong or maintain the activity of microorganisms.