Effects Of Water Soluble Alfalfa Polysaccharides On Growth Performance And Immune Functions Of Broilers

Polysaccharides were the class of common natural macromolecules in animals, plants, microorganisms, algae and other organisms, were the major component of living organisms. Studies had proven that Polysaccharides could regulate immune function, antimicrobial, antitumor, antiviral, antioxidant, antithrombotic, anticoagulant, decrease lipid and other functions. According to the source, Polysaccharides could be divided into plants polysaccharides, fungi polysaccharides, algal polysaccharides, animal polysaccharides, bacterial polysaccharides. A large number of tests showed that plant polysaccharides have a wide range of biological activities, widespread immune function. This research on the basis of plant polysaccharide-Water Soluble Alfalfa Polysaccharides (WSAP), extracted in the laboratory and factory, by supplement in ration or in vitro lymphocyte culture, growth performance and immune function were studied.Experiment 1: Extract and Determination Water Soluble Alfalfa PolysaccharidesTo investigate the difference between extraction ratio and content of Water Soluble Alfalfa Polysaccharides (WSAP) extracted in different environments, provide scientific basis for biological functions and the industrial production of WSAP. WSAP were extracted in laboratory and factory. (1) In the laboratory, extracting pressure were 120 ~ 150kpa, water extraction 2 times, each was 1h, the two extraction solid-to- liquid ratio were 1:3 and 1:2 respectively; in the factory extracted 2 times, each time was 1h, the two extraction solid-to- liquid ratio were 1:8 and 1:6 respectively. (2) In laboratory ,the extraction ratio and conditions of WSAP were significantly higher than those in factories, in laboratory WSAP extraction ratio was 10.75%, in factory was 5.72%; the content of WSAP was 82.13% in lab extration, 45.84% in factory.Analysis the two WSAP extraction ratio and content, the difference may be due to (1) under laboratory conditions and high raw material grinding may cause a higher extraction rate of polysaccharides; In addition, in the factory a large number of water extraction liquid extraction with the residue loss, which may be the important reason to cause WSAP for low extraction ration. (2)In factory , groundwater in the high mineral content, so the crude ash content of WSAP in factory(29.49% ) were higher than in the laboratory (6.24%). (3) Deproteinized in Laboratory were accurate to judge the end, the crude protein content (5.99%) is lower than in factory extraction(13.98%). These results suggest that polysaccharides extraction ratio and content could be difference in differet extraction conditions, we should not only consider the extraction and concentration of the extraction step, the processing of raw materials, extraction reagents, and the rational use of extraction devices should also be studied.Experiment 2: Effects of supplemental Water Soluble Alfalfa Polysaccharides on growth performance and immune functions of Broilers This experiment was conducted to study the effects of Water Soluble Alfalfa Polysaccharides (WSAP) added in ration on growth performance and immune properties of broilers. A total of 240 1-day-old broilers were randomly assigned to eight groups with 3 replicates of 10 broilers each. Test is divided into two phases, 0 ~ 3 weeks for the pre-phase trial, 4~ 6 weeks for the last-phase trial. (1) In this experiment,200mg/kg, 500 mg/kg WSAP groups average body weight and daily gain were significantly higher (P <0.05), feed gain ratio was significantly lower than the control group (P <0.05). (2) A certain concentration of WSAP could significantly promote the immune organs index of broilers HI, IgG and peripheral blood T, B lymphocyte proliferation (P<0.05), and the optimum amount is 500mg/kg. The results show that a certain concentration of WSAP could promote the growth performance and immune function of broilers, the optimum amount is 500mg/kg.Experiment 3:Effects of Water Soluble Alfalfa Polysaccharides on cell proliferation of broiler lymphocytes in vitro .To investigate the effects of Water Soluble Alfalfa Polysaccharides (WSAP) on cell proliferation of broiler lymphocytes in vitro . Different concentrations of WSAP were added into cultured with ConA stimulated chicken peripheral blood, thymus, spleen lymphocytes and into cultured with LPS stimulated chicken peripheral blood, spleen, bursa of fabricius, cultured for 48 hours, use MTT colorimetric determination of lymphocyte proliferation. (1) In this experiment, a certain concentration of WSAP groups (4μg/mL~20μg/mL) OD values were higher than control groups, with the polysaccharide concentration increased, OD values increased first (1μg/mL~20μg/mL) and then decreased (100μg/mL~200μg/mL). (2) The same concentration WSAP on broiler peripheral blood and other immune organs, the stimulus intensity were different, peripheral blood and spleen T, B lymphocytes OD values were less than the thymus and bursa lymphocytes OD values. (3) The best stimulus concentration for peripheral blood and other organs were difference, for peripheral blood was 20μg/mL, for spleen was 8μg/mL, for bursa and thymus were 15μg/mL. In this experiment, the concentration of WSAP were 8μg/mL, 15μg/mL, 20μg/mL could promote the broiler lymphocyte proliferation, which may be the one of the mechanisms that polysaccharides could enhance immune function.Experiment 4:Effects of Water Soluble Alfalfa Polysaccharides on Lymphocyte immune regulation effects of broilersTo explore WSAP on immunization of broiler lymphocyte cultured in vitro, studied on lymphocyte chemiluminescence, NO, IL-2, IL-6 and IFN-γ. Different concentrations of WSAP added into peripheral broiler, thymus, spleen, bursa lymphocytes, with bio-chemiluminescence measurement lymphocytes chemiluminescence value; cultured for 24 hours, NO content of lymphocytes secretion would be determined with nitrate reductase; different concentration of WSAP respectively added into the peripheral broiler, thymus, spleen lymphocytes in vitro, detected Cytokines IL-2,IL-6,IFN-γon Elisa. Results show that (1) a certain concentration WSAP could enhance peripheral broiler, spleen, thymus and Bursa lymphocytes chemiluminescence, could promote the expression of lymphocyte NOS, improve iNOS activity and promote the production of NO; could stimulate peripheral broiler, spleen, thymus lymphocytes secreted IL-2, IL-6 and IFN-γ, with WSAP concentration, values increased first (1μg/mL~20μg/mL) and then decreased (50μg/mL~200μg/mL); (2) the best stimulus concentration with different WSAP on tissues and organs were various, the best stimulation on peripheral blood of WSAP concentration were 15 and 20μg/mL, spleen was 8μg/ mL, thymus was 15μg/mL;(3) broilers of the 21-day old, the lymphocytes section of NO quantities, NOS activity, IL-2, IL-6 content and the sensitivity with the WSAPwere exceed than 42-day old broilers, but IFN-γwere contrary.Comprehensive analysis, concentration of WSAP on 8, 15, 20μg/mL, could promote broiler lymphocytes chemiluminescence significantly, could promote the expression of lymphocyte NOS, improve iNOS activity and promoted the production of NO, can stimulate peripheral broiler, spleen, thymus lymphocytes secretion IL-2, IL-6 and IFN-γ, approached to regulation of broiler lymphocyte proliferation.