【作者】 刘瑞生；

【导师】 魏彦明； 薛掌林；

【作者基本信息】 甘肃农业大学 ， 预防兽医， 2005， 硕士

【摘要】 黄芪多糖（APS）是中药黄芪的主要活性成分之一,具有促进抗体生成,诱导干扰素产生和促进免疫反应等作用。APS主要以注射剂应用,免疫期短。而近年来非典型性鸡新城疫（ND）不断发生,虽然采用疫苗多次预防注射仍不能控制该病。因此有必要研制开发能提高鸡免疫力,增强疫苗免疫效果,维持时间长的APS新剂型,以达到控制该病的目的。采用水提取、CaO水溶液提取和Na₂CO₃水溶液提取法从黄芪中提取APS,计算收率和成本,用苯酚-硫酸法测定APS含量。结果表明CaO水溶液提取法APS收率和含量极显著高于水提取和Na₂CO₃水溶液提取法（p＜0.01）,其提取成本极显著低于水提取法（p＜0.01）,显著低于Na₂CO₃水溶液提取法（p＜0.05）。通过肉鸡ND免疫试验,建立了诊断鸡免疫功能变化的检测方法—血凝抑制试验、T淋巴细胞-E玫瑰花环试验和酸性α-醋酸萘酯酶测定试验。对7日龄肉用雏鸡和蛋雏鸡进行鸡NDⅣ系苗、油乳剂苗、Ⅱ系苗和Ⅰ系苗（30日龄）免疫同时,配合应用不同剂量的APS溶液饮水、与油乳剂苗混合或分别皮下注射、作为稀释液稀释鸡NDⅡ系苗后点眼滴鼻、与NDⅠ系苗混合或分别肌肉注射,通过测定比较免疫后10、20和30d抗体水平变化,发现添加APS20mg/只对鸡ND不同种类疫苗免疫增强作用效果明显,抗体水平高,为APS最佳剂量。以20mg/只APS分别制成不同剂型供鸡饮水、皮下注射、肌肉注射和点眼滴鼻,在雏鸡7日龄注射鸡NDIV疫苗时配合应用,结果免疫后10、20和30d使用APS不同剂型的雏鸡抗体水平显著高于对照组（p＜0.05）,但试验组间差异不显著（p＞0.05）。APS饮水使用方便,避免抓鸡造成应激,节省人力,因此为APS最佳使用剂型。急性和慢性毒性试验表明,APS对鸡安全,无毒副作用。效力试验发现APS可以提高鸡T淋巴细胞E-花环形成率和B淋巴细胞花环率,表明APS可以提高鸡的细胞免疫功能和体液免疫功能。经过27000只肉用雏鸡和蛋雏鸡田间试验和区域试验表明,雏鸡进行鸡ND疫苗免疫时,配合2%APS溶液饮水,可以增强机体免疫功能,延长疫苗免疫保护期,提高鸡成活率,降低发病率和死亡率,具有显著的经济效益。更多还原

【Abstract】 The Astragalus Polysaccharide（APS）is one of main active constituents of Chinese herbal Astragal that functions in the promoting antibody formation,and the inducibility of interferon produces,improving immune response.APS had been mainly applied to injection purpose,and gotten short time for immunity.However,non-typical Newcastle Disease（ND）occurred unceasingly in recent years,which had been still not to be able to be controlled although using the vaccine many times.Therefore it is necessary to develop new type APS to be used to enhance the chicken immunity,and the vaccine immunity effect,to effectively control this disease.APS extracted from Chinese herbal Astragalus with the aqueous extract,CaO aqueous extract and Na₂CO₃ aqueous extract respectively were computed the receiving rate and cost,and determined the APS content with the phenol-sulfuric acid method.The results indicated that the APS extraction rate and the content of CaO aqueous extract were remarkable higher than the aqueous extract and Na₂CO₃ aqueous extract（p＜0.01）.The cost of CaO aqueous extract was remarkable lower than the aqueous extract（p＜0.01）and Na₂CO₃ aqueous extract（p＜0.05）.The diagnosis chicken immunity function change Monitor method-hem agglutination inhibition test,thymus dependent lymphocyte erythrocyte rosettes assay and the acida-naphthy acetate esterase determination experiment were established through the broiler ND immunizing experiment.While respectively immune broiler and young layer which were aged 7 days with NDⅣvaccine,oil-emulsion vaccine,Ⅱvaccine andⅠvaccine（at the age of 30 days）,the coordinate use different dosage APS solution to drink water,with the oil-emulsion vaccine,mix or the distinction hypodermic injection,as a diluent to dilute NDⅡvaccine,then to drop eyes and nose,with NDⅠvaccine mix or the respectively intramuscular injection,through determining and comparing immune function changes on 10、20 and 30 days after immunity,it was discovered that increase 20mg/only APS was the best dosage,It indicated that enhanced function effect obviously to different type vaccine immunity of chicken ND,and made the antibodies level higher.Different forms of prepared APS with only 20mg per chicken for drinking, hypodermic injection,and intranmseular injection,drop eyes and nose as chicken were injectedⅣvaccine of chicken ND at 7 days.The results showed that the immune body level of test groups were higher than the control groups after using different forms of APS on 10,20 and 30 days,but the test groups were not responded remarkably.The experiment showed that the APS was beneficial to drink with water,avoid catching stress,save the manpower,so was the best form.Acute and chronic toxicity tests also indicated that APS was safe to chicken,there were no any toxic and side effect.The potency test showed that APS might enhance thymus dependent lymphocyte E-rosette formation rate and bursa dependent lymphocyte formation rate.It indicated that APS might enhance cell immunity function and humeral immunity function of chickens.The field test and regional test through 27000 chicken and young layers indicated that chickens were well immune with ND vaccine around 3 days at the age of 7 days,meantime with drinking 2%APS solution might strengthen organism immunity function,reduced the incidence and death rate.It had remarkable economic efficiency.更多还原