Effect Of Plantain Polysaccharides On Immune Efficacy Of ND Vaccinated Chickens

In recent years, more and more infectious disease especially virus disease of livestock and poultry were serious harm to the animal husbandry industry, vaccination is the most important control measures. Many of the vaccines rely on adjuvant to enhance the protective response. Adjuvant can enhance the immune response, or change the immune response type of material, non immunogenicity when used alone in general. The role of adjuvant in immunity is mainly to enhance the immune response, including enhance immune responses more Longer and more lasting. In recent years, the study found that the plant polysaccharide has adjuvant effect, can enhance the immune effect of the vaccine, promote the specific immunity and nonspecific immunity, cellular immunity and humoral immunity. Plant polysaccharide adjuvant has the advantages of natural, low toxicity, no drug residues, has an advantage in the use of safety. But at present, which can be used in the practice of plant polysaccharide adjuvant is not much, to find a rich resource of plant polysaccharide adjuvant in order to meet the need of practice is imminent. Plantain is widely distributed all over the world, there are three genera and about 270 species, in our country only has Plantago plantain. Plantain is widely distributed in China, is a kind of traditional Chinese medicine, has the effects of clearing heat, cooling blood, detoxification, diuretic and so on. Modern pharmacology study shows, plantain contains rich biological active components, such as polysaccharides, flavonoids, ursolic acid and glycoside etc. Polysaccharide is the main material of plantain exert biological activity. Bent (1995) study of plantain polysaccharide found it has L-Arabia sugar (20%), D-galactose (28%), D-glucose (6%), D-Mannose (2%) L-mannose, rhamnose (4%), D-glucose acid (31%) and a small amount of D-xylose and fucose carbon, With B-1, 4 are connected to a main chain, side chain containing 2 and 3. The research of plantain polysaccharide as immune enhancement agent is very little. This study uses of plantain polysaccharide as adjuvant Newcastle disease vaccine, to find of plantain polysaccharide enhance the immune effects.1. The study of Plantago polysaccharide extraction and purificationUsing ultrasonic extraction method extraction of plantain polysaccharide. determine the factors and levels by the single factor experiment and uniform design experiment. The best extraction process was as follows:ultrasonic time 60min, temperature 60 degrees Celsius, the ratio of solid to liquid 1:30, the extraction rate was 75.88mg/g. The extracted crude plantain polysaccharides were purified, removal of protein by three trichloroacetic acid method, protein removal rate was 52.1%, by active carbon, decoloration rate was 82.3%. Analysis on the physical and chemical properties: plantain polysaccharide is yellow brown powder; soluble in water, dilute acid, dilute alkali; insoluble in ethanol, petroleum ether, acetone, butanol and other organic solvents containing; reducing sugar; no starch, Monosaccharide and two sugar; Plantago polysaccharide solution PH 7.8. Plantago polysaccharide has the in vitro scavenging free radicals effect, Plantago polysaccharide concentration of 70 g/ml of OH-scavenging rate was 41.1%; the concentration of 125 g/ml of O2·- clearance rate was 37.2%; the concentration of 160 g/mll ofNO2·- clearance rate was 72.5%.2. The effects of plantain polysaccharide on growth, physiological and biochemical indexes and immune organ indexGiven the Plantago polysaccharide solution each time per chicken for six consecutive days, and given the same amount of saline solution for control. And sampling on the 5(D5),10 (D10).15 (D15),20 (D20).28 (D28) days after administration, determination of weight gain, blood routine, biochemical and physiological indexes, immune organ index and Observation of the immune organs and tissues.The results showed that:The weight in plantain polysaccharide group’s chickens were 214.35g、359.71g、633.44g、826.52g on the D10、D15、D20. D28 days, were higher than the that of control group’s chickens, extremely significantly (p＜0.01); on D5 to D15 day plantain polysaccharide group RBC (1.84～2.45×1012/L) was significantly higher than that of control group RBC (1.40～2.14×1012/L), (P＜0.05); on D28 day plantain polysaccharide group WBC (258.8×109/L) was significantly higher than that of control group WBC(128.1×109/L), (P<0.05); on D10～28 day plantain polysaccharide group GOT (131.9～27.57IU/L) and GPT (8.53～ 16.99IU/L) was higher than that of control group GOT (27.2～.67IU/L) andGPT (2.58～ 5.87IU/L), (P<0.05, P＜0.01); on D5～28 day plantain polysaccharide group AKP (10.47～ 10.15IU/L)and PCa(1.59～1.01 mmol/ml)was no significant difference t compared to control group AKP (7.54-3.4IU/L) and PCa (1.31-1.07 mmol/ml) (P> 0.05); on D5-D28 day plantain polysaccharide group thymus index (2.04±0.28～2.93±0.23)、spleen index (1.2±0.14～1.90±0.12) and Bursa of fabricius index (4.11±0.46～5.98±0.25) was higher than that of control group thymus index (1.38±0.24～2.40±0.31)、spleen index (1.06±0.32～1.52±0.28) and Bursa of fabricius index (2.54±0.77～.47±0.29), (P＜0.05, P＜0.01); on D5～D20 day plantago polysaccharide group of chicken red pulp of spleen lymphocytes, plasma cells and macrophages were more than that in the control group, on D10～D20 day thymic lobule area of plantain polysaccharide group were higher than that of control group, on D10～D28 day plantago polysaccharide group bursal sections were significantly higher than control group.The results shows that the plantain polysaccharide can promote the growth of chickens to promote weight gain, organ development and improving immunity.3. The effect of plantain polysaccharide on immune efficacy of Newcastle disease vaccine320 one-day-old unvaccinated chickens were randomly divided into seven groups, which were Newcastle disease (ND) vaccine group、ND vaccine+Plantago polysaccharide (50mg/ml) group、 ND vaccine+Plantago polysaccharide (25mg/ml) group、ND+Plantago polysaccharide vaccine (12.5mg/ml) group、ND vaccine+Astragalus Polysaccharide (1.5g/ml) group、ND+dexamethasone (group 2.5mg/ml) vaccine and the healthy control group. Sampling on the 7 (D7)、14 (D14)、21 (D21)、35 (D35) days after the first injection, determination of ratio of Viscera index, hemagglutination inhibition antibody titers. peripheral blood T lymphocyte proliferation rate and peripheral blood T lymphocyte CD4+/CD8-. The index of immune organs results showed that: Plantago polysaccharide 50mg/ml dose group, the thymus index (6.45+0.24 to 3.07+0.17), spleen index (3.12+0.11 to 2.05+/-0.11) and the index of bursa of Fabricius (7.66+0.09 to 3.31+0.13) and astragalus polysaccharide group thymus index (6.07+0.56 to 3.24+0.22), spleen index (3.31+ 0.42 to 2.11+0.16) and the index of bursa of Fabricius (8.06+0.09 to 3.45+0.16) was significantly or highly significantly higher than that of thymus index ND immune group (4.23+0.13 to 2.77+ 0.12), spleen index (2.28+0.11 to 1.83+/-0.12) and the index of bursa of Fabricius (from 0.45 to 5.86.2.97+0.12), (P< 0.05, P< 0.01). The antibody titer results showed that:Plantago polysaccharide 50mg/ml (11.25～9.50),25mg/ml (10.25 to 8.75),12.5mg/ml (10～8.50) dose group and astragalus polysaccharide group (11.25～9.50) from D14 to D28 was significantly higher than ND group (8.75～6.75) (P< 0.01), dexamethasone group (5.33; 5) in the D14-D35 is lower than that of ND group significantly (8.75～6.75) (P< 0.01). The rate of lymphocyte transformation results showed that:Plantain 50mg/ml dose group (2.1733+0.042 to 2.0623+0.061) and astragalus polysaccharide group (2.0678+0.059 to 2.0267+0.099) D7-D28, T lymphocyte transformation rate were significantly higher than that in the ND vaccine group (1.8967+0.031 to 1.7213+0.084). (P< 0.01); dexamethasone (1.5633+0.045 to 1.5723+0.045) in the D7-D21 below the ND immune group significantly (1.8967+0.031 to 1.7213+0.084) (P< 0.01). The the ratio of CD4+/CD8+results showed that:Plantain polysaccharide 50mg/ml dose group (1.9186+0.035 to 1.8013+0.028) was higher than that in ND vaccine group D7-D35 ratio of CD4+/CD8+ significantly(1.6417+0.02 to 1.3507+0026),(P<0.01,P<0.05)；Plantago polysaccharide 25mg/ml(2.0347+/.0.035 to 1.7573+0.028),12.5mg/ml group(2.070+0.061 to 1.7487+0.02) and astragalus polysaccharide group(2.0263+0.043 to 1.7620+0.03)D14～D35 in the ratio of CD4+/CD8+was significantly higher or significantly higher than that of ND vaccine group(1.5867 +0.04 to 1.3507+0026),(P<0.01.P<0.05):Plantago polysaccharide 50mg/ml dose group(1.9186 +0.035 to 2.0697+0.036)D7～D21 ratio in CD4+/CD8+was significantly higher than that of Astragalus polysaccharide group(1.7710+0.006 to 1.7593+0.05),(P<0.05)；dexamethasone group in D7一D35 days was significantly lower than vaccine group(1.6417+/-0.02 ND-1.3507+ 0026),(P<0.01).The results showed that plantain polysaccharide can enhance the inmune effect of ND vaccine.