Effects Of Wall-Broken Bee Pollen, Beer Yeast And Yeast Polysaccharides Extracted From Beer Yeast On Layers' Immune Function

To develop an immune intersifier, beer yeast and bee pollen of pagodatree flower was scrapped as raw materials, polysaccharides of cell-wall was selected from wall-broken beer yeast by extraction of alkalescence solution and was analysed.Then the three substances respectively wre added to layer's feed with different doses.By detecting their influences on the specific and non-specific immunoreaction of layers, their value could be primarily identified as immune intensifiers.The contents which have been done were as follows:1. Effects of wall-broken been pollen on the immune function andintestinal bacterial flora of layers.Bee pollen of pagodatree flower were scrapped by ultra microcom minution with vibrant-mill, and used to feed layers from 7 days to 50 days with three different doses, 1g/kg , 5g/kg , 10g/kg .Each dose were applied to two experimental groups.In the first group,layers wre immuned with ND-LaSota,the dynamic changes of HI antibody titer were detected in the serum with B-micromethod of HI. In the second group layers were immuned with the emulsion inactivated vaccine of E.coli, the changes of the antibody titer of E.coli were detected with indirect HA. The spleen , the chest gland , the bursa of fabricius were regularly selected. The indexes of the immune organs were caculated. The bacteriolozical examinations of E.coli and lactic acid bacteria were done with intestinal contents, at the same time , pH was tested in the intestine.The result showed that to some degree,different doses of wall-broken bee pollen can improve the antibody titer of viral antigen andbacteroidal antigen.Layers organ growth,intestinal environment and the balance of intestinal bacterial flora had also been influenced to some extent.2. Effects of wall-broken beer yeast on the specific and non-specificimmunoreaction of layerThe young-layers were randomly allocated to five groups, fed with five treatment diets supplemented with different doses of 100g/kg^ 50g/kg> 10g/kg> 5g/kg ^ lg/kg respectively, and immuned with ND-LaSota vaccine and the emulsion inactivated vaccine of E.coli. The layers were selected blood regularly. The blood were devided into groups,group one were coagulated, and group two were heparinize to anti-coagulation. The serum from group one could be used to detect antibody titer of ND with P-micromethod of HI and with indirect HA.The lymphocytes were separated with lymphocytes sepatation medium(LSM) from blood of group two, and ANAE+ lymphocyte were calculated with a- naphthalene esterase (ANAE) test to observe the influence of wall-broken yeast on cell immunity.Regularly, the spleen , the chest gland , the bursa of fabricius were weighed and used to calculate the immune indexes to observe the influence of wall-broken yeast on the growth of the young layers' immune organs. The result showed that there were obvious influences on the ND antibody titer and E.coli antibody titer of the serum with different doses of wall-broken yeast.The indexes of immune organ, antibody decrease and increase, the ratio of ANAE+ lymphocyte were related to the added doses. The added doses had obvious difference when the number of E.coli and lactic acid bacteria was different in intestinal contents.3. The discussion of the extractive technique of the polysaccharides ofyeast and the analysies of its components.The polysaccharides of yeast were selected from beer yeast with ultra microcom minution cell-wall and by extraction of alkalescence solution. The cell of beer yeast could be obtained to be wiped off impurity and bitter andultra microcom minution cell-wall.A buff powder could be gotten by extraction method, alcoholization and freeze drying.The influences of the three main factors: time, ratio of material and water, temperature on the production rate of polysarrides were mainly studied.The content of the polysaccharides of yeast were determinated with phenol-sulfuric acid colorimety. The components of polysaccharides of yeast were analysed by thin layer chromatography. The result showed that, when the temperature was 100°C, the ratio of material and water was 1:10, select-time was 3 hours, the rate of beer yeast polysarrides selection could be gotten 10.21%. The protein content of extraction beer yeast polysarrides reached as high as 10.32%, with a relative purity of 78.46%. Therefore, the selection were yeast polysarrides which contained protein. It could be found that yeast polysarrides were the mixture of poly-mannose and poly-glucose by thin layer chromatography.4. Impact of beer yeast polysaccharides on layers' humoral immunity andintestinal bacterial floraThe yeast polysaccharides of different doses were added to the feed . And young-layers were immuned with ND LaSota and the emulsion inactivated vaccine of E.coli. The layers were selected blood regularly. The dynamic changes of antibody titer of ND and E.coli were tested with P-micromethod of HI and with indirect HA. The E.coli and lactic acid bacteria in intestinal contents of different parts were counted.Results showed that, the influence of tiny dose (lg/kg) and high dose (50g/kg) on antibody titer was not obvious (P >0.05 ), while the low dose (5g/kg) and medial dose (lOg/kg) could obviously improve the antibody titer(P <0.05 ); the counting results showed that the influence of the tiny dose (lg/kg) and high dose (50g/kg) on the intestinal contents was not obvious(P >0.05 ), while the low dose (5g/kg) and medial dose (lOg/kg) could obviously increase the number of lactic acid bacteria in the intestinal contents(P <0.05 ). However, the influence of different doses on the number of E.coli was not obvious (P >0.05 ).It could be concluded that layers which were immuned with ND vaccine and E.coli vaccine, antibody titer could be increased because of adding different material. The intestinal environment was improved when the proper doses of wall-broken bee pollen, beer yeast and extracting polysaccharides form yeast were added to the feed.At the same time , results showed that the selected beer yeast polysaccharides still had better biology-activity by extraction of alkalescence solution, The beer yeast polysaccharides not only enhanced layer's humoral immunity, but also improve the intestine environment.Therefore, been pollen and beer yeast could be widely used as immune intersifier.However, diverse polysaccharides had different compotent and structure, then there were discrepancy in biology-activity. Futher study should be done about biology-activity of bee pollen, structure analysis of yeast polysaccharides on the molecular degree and intestinal intestinal bacterial flora growth mechanism.