Researching And Producing On Feed Compound Probiotics Of Bacillus

As the food safety awareness of people continuing to enhance, microecologics instead of antibiotics as feed additives have been widely recognized by feed industry, with its natural, safe, non-reside and other adventage. Because of the special characteristic of natural strong stress resistance, high temperature tolerance, acid and alkali tolerance, and easy storage, probiotics of bacillus became a research focus at home and abroad. This study developed subtilis-lichens-natto triple bacillus viable preparation through experiments, provide a theoretical basis and technical support for the industrial production of probiotics and the application in raising of weaned piglet.This study combined 3 strain of bacillus of subtilis(B.S),lichens(D.Y) and natto(N.T) to dual culture, obtained triple bacillus, its feed enzyme activities were:protease 126.72 U/mL, amylase 31.31 U/mL, cellulase 1.77 U/mL,β-mannase 2.42 U/mL. Identify of colonial morphology,cell characteristics,physiological and biochemical characteristics of 3 strains were conducted. Through orthogonal and single factor experiment, obtained optimum medium composition ratio:1%(v/v) waste molasses,0.5%(w/v) soybean powder,0.3%(w/v) calcium chloride. The favorite fermentation conditions:initiative pH7.5, inoculation amount5%(v/v), liquid volume 10～30%(v/v), culture temperature34℃, and ferment time 20 h. Utilizing industrial residues molasses,soybean powder as carbon,nitrogen, fundamentally reduced the cost of production. Based on the optimized medium with 5%(w/v) wheat bran and in standing fermentation liquor after the end of culture for 48 h, the sporulation rate of triple bacillus was increased to 96.2% or more. Through test, the triple bacillus preparation of high-temperature and simulated pig gastrointestinal solution was well tolerated, under a protective agent[5%(w/v) glucose],-7℃(frozen) condition, after 90 d the number of viable count remained at 109 cfu/g level, the survival rate up to 21.49%. Triple bacillus by 5 t level of pilot scale fermentation production, the viable count of bacterial sludge after centrifugation as high as 9.0×1012 cfu/g, the viable cells of preparation were 4.5×1010 cfu/g. The results of feeding trial of piglets showed that triple bacillus probiotics can make piglets daily weight raised by 20.72% (P>0.05), feed-weight ratio reduced by 12.36%(P<0.05), piglets diarrhea rate reduced by 22.73%(P>0.05), feed efficiency increased, economic efficiency enhanced significantly.From the above, the subtilis-lichens-natto triple bacillus viable preparation prepared by this study have low cost,favorable stability and high economic efficiency, as feed additive in piglet production will appear very broad application prospects.