青刺果种粕粉营养成分分析及其对肉鸡的免疫调节作用研究
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摘要
营养物质与机体免疫之间的关系逐渐被揭示,营养物质不但为动物维持正常的生长发育所必需,而且是维持免疫系统的功能并使免疫活性得到充分表达的决定性因素。因此,营养免疫学也逐渐成为人们研究的热点。随着营养免疫学的逐渐发展,人们更把注意力转向了大自然中的草本植物,因为它所提供的均衡、纯净的营养是提升免疫功能的最佳能源。通过大量的资料证明,青刺果(Prinsepia Utilis Royle)这种植物含有水分、灰分、粗纤维、粗脂肪、粗蛋白、17种氨基酸、维生素A、D、E、K以及多种微量元素等丰富的营养物质。在我国,青刺果资源丰富,这为研究、开发、利用青刺果提供了良好的条件。然而长期以来,对青刺果的研究主要集中在青刺果的种植,对青刺果的开发利用也集中在青刺果油上,而对青刺果压榨青刺果油后的种粕粉的开发利用处于空白状态。因此,本研究拟通过对青刺果种粕粉中的氨基酸、粗蛋白、粗脂肪、维生素、微量元素等营养成分进行检测分析以及动物试验,进而探讨青刺果种粕粉对肉鸡免疫调节作用。主要研究内容如下:
1.青刺果种粕粉营养成分分析本试验首次对青刺果种粕粉进行营养成分的检测,其中,青刺果种粕粉中的氨基酸组分采用氨基酸自动分析仪测定;粗蛋白采用凯氏定氮法;粗脂肪采用索氏抽提法测定;维生素A采用高效液相色谱法;微量元素采用原子吸收光谱法测定。测定结果显示,青刺果种粕粉中各种营养成分的含量如下:氨基酸36.83%;粗蛋白41.26%;粗脂肪6.12%,维生素A 35.40mg╱g,微量元素含量分别为Fe 122.29 ug/g、Zn 54.64 ug/g、Mg 2.47ug/g、Mn 35.01 ug/g、Cu 9.38 ug/g。结果表明青刺果种粕粉中含有丰富的与免疫相关的营养成分。
2.青刺果种粕粉对肉鸡免疫调节作用的研究动物试验:将60只1日龄雏肉鸡随机分为3组,每组20只。Ⅰ组为对照组,饲喂基础日粮;Ⅱ组和Ⅲ组7日龄开始分别在基础日粮中添加10g/kg和15g/kg青刺果种粕粉,将增重最好的组作为试验组。在7、14、21和28日龄采取增重效果最好组采集血样和免疫组织材料(脾脏、法氏囊、胸腺)。血样用于测定血清抗体IgG(Immunoglobulin G)、IgA(Immunoglobulin A)、IgM(Immunoglobulin M)、谷草转氨酶GOT(Glutamic Oxalacetic Transaminase)、谷丙转氨酶GPT(Glutamic Pyruvic Transaminase)、血清钙、血清白蛋白SA(Serum Aibumin)、血清碱性磷酸酶AKP(Alkaline Phosphatase),以及红细胞C3b受体花环率E-C3bRR(Erythrocyte C3b Receptor Ratio)、红细胞免疫复合物花环率E-ICR(Erythro- cyte immune Compound Ratio)、总淋巴细胞玫瑰花环率Et(T-lymphocyteErthrocyte Total Rosette Rates)、活性淋巴细胞玫瑰花环率Ea(T-lymphocyte Erthrocyte ActiveRosette Rates)、T淋巴细胞百分率。免疫组织材料用于测定脏器指数、免疫器官中T淋巴细胞和巨噬细胞数量,同时制作石蜡切片,苏木素伊红HE(Hematoxylin and Eosin Staining)染色后观察免疫器官的组织学结构变化;脾脏组织制作电镜切片,电镜下观察脾脏超微结构的变化。结果显示:
(1)青刺果种粕粉对肉鸡血清抗体及血液生化指标的影响表现为:青刺果种粕粉10g/kg剂量组与对照组相比,IgG在各日龄都表现为显著升高;IgA差异不显著,但与对照组相比均有所提高;IgM在21日龄时差异显著,表现为显著升高,在其它日龄均有所提高。GOT、GPT和AKP在14日龄时,与对照组相比差异显著升高,但随着试验的进程,逐渐降低至正常;SA和钙的含量与对照组相比,差异不显著。
(2)青刺果种粕粉对肉鸡红细胞免疫及T淋巴细胞的影响表现为:青刺果种粕粉10g/kg剂量组与对照组相比,E-C_(3b)RR率和E-ICR率差异不显著;T淋巴细胞ANAE~+率与E-玫瑰花环率在各日龄均较对照组极显著升高(P＜0.01)。
(3)青刺果种粕粉对肉鸡免疫器官中免疫细胞的影响表现为:试验组各免疫器官中的T淋巴细胞数量的含量与对照组相比,主要表现在试验初期(14、21日龄),差异显著(P＜0.05或P＜0.01),表现为不同程度的升高;试验组各免疫器官中的巨噬细胞数量与对照组相比较,主要表现在试验后期(21、28日龄),差异显著,表现为明显升高。
(4)青刺果种粕粉对肉鸡免疫器官发育影响的结果表明,试验组肉鸡法氏囊、胸腺、脾脏脏器指数在各个日龄均显著或极显著高于对照组(P＜0.05或P＜0.01)。光镜下,14日龄时,试验组较对照组法氏囊皮质与髓质分界较清楚,淋巴滤泡形成,胸腺皮质不同程度增厚,脾脏形成动脉周围淋巴鞘;21日龄时,试验组较对照组法氏囊中淋巴细胞发育成熟,形态较小,淋巴滤泡直径增加,胸腺小体增多,皮质增厚,脾脏淋巴细胞致密;28日龄时,试验组与对照组比较,法氏囊淋巴滤泡较大,胸腺小体增大,可见朗罕氏细胞,脾小体和生发中心较发达。电镜下,试验组脾脏淋巴细胞形态均一,核仁增多,核染色质边移,随着日龄的增长,淋巴细胞活性增加。
试验结果证明,青刺果种粕粉对肉鸡的血清生理生化指标无不良影响;能提高血清抗体含量,对的细胞免疫具有积极的调节作用;能促进肉鸡免疫器官的生长发育成熟。由此可见,青刺果种粕粉含有肉鸡生长发育所需要的各种营养物质,对肉鸡的早期生长发育具有免疫调节作用。
The relationship between the immune and nutrition gradually being revealed,Not only for animal nutrition to maintain normal growth and development necessary,but also to maintain the immune system function and the decisive factor is immune activity is fully expressed.Therefore,the nutritional immunology research have gradually become the hot spot.With the nutritional immunology to development gradually,people to turn their attention to the nature of herbaceous plants,Because it provides a balanced,pure nutrition is the best energy to enhance immune function. Through a lot of information that Prinsepia Utilis Royle contains rich nutrients such as water,ash, crude fiber,fat,protein,17 kinds of amino acids,vitamins A,D,E,K,and a variety of microelement. In China,Prinsepia Utilis Royle is rich in resources,it provided a good condition.for research, development and use.But a long time,after Prinsepia Utilis Royle squeezed the Prinsepia Utilis Royle oil,the Prinsepia Utilis Royle Fruit Meal blank in the development and utilization of state.So, study the immunoloregulation effects of Prinsepia Utilis Royle on chicken,by the animal experiment and the nutritional components analysis of Prinsepia Utilis Royle Fruit Meal such as amino acid,crude protein,crude fat,vitamin A and microelement.The main contents are as follows:
1.Analysis of the Nutrition Ingredient of Prinsepia Utilis Royle Fruit Meal Analysis of the Nutrition Ingredient of Prinsepia Utilis Royle Fruit Meal at first time in this experiment.Analysis of the amino acid、crude protein、crude fat、vitamin A and the microelement by the methods of the amino acid autoanalyser、Kjeldah method、Soxhlet、high efficiency liquid chromatography and atomic absorption spectrotype,respectively.The results showed that the Prinsepia Utilis Royle Fruit Meal was a good material with rich nutrients,the content of amino acids is 36.83%,36.83%,the content of crude protein is41.26%,the content of crude fat is 6.12%,the content of vitamin A is 35.40mg/g,the content of microelement Fe、Zn、Mg、Mn、Cu is 122.29 ug/g、54.64 ug/g、2.47 ug/g、35.01 ug/g、9.38 ug/g.The results prove that the Prinsepia Utilis Royle Fruit Meal is rich in nutrition which has immune-related components.;
2.The Immunoloregulative Effectsof Prinsepia Utilis Royle Fruit Meal on Chicken Animal experiment:Sixty 1-day-old chicken were divided into three groups randomly.Group I feeded with basal diet.GroupⅡand groupⅢwas supplemented with 10g/kg and 15g/kg Prinsepia Utilis Royle Fruit Meal in the basal diet respectively.After 14d、21d and 28d the group which effect of weighting is best was picked out to blood and immune organize(thymus,spleen,bursa)sampling.Blood be use to mensurate erythrocyte C3b receptor ratio(E-C3bRR)、erythro- cyte immune compound ratio (E-ICR)、Et、Ea、percent of T lymphocytes、Antibody level(IgG、IgA、IgM)and Blood Biochemical Indicator(Glutamic Oxalacetic Transaminase(GOT),Glutamic Pyruvic Transaminase(GPT),serum calcium,Alkaline Phosphatase(AKP),Seralbumin(SA)).Immune organize be used to mensurate the index of immune organ,the quantity of T lymphocyte and macrophage.And then we detected change of structure after making splices and Hematoxylin and Eosin(HE)staining.Otherwise ultrastructure of spleen was be observed under electron microscope.
The results showed that(1)the effects of Prinsepia Utilis Royle Fruit Meal on the sera antibody and blood biochemical indicator in chickens as flow:Comparable with control group,The content of IgG increased significantly after taken Prinsepia Utilis Royle Fruit Meal.The content of IgA had no significantly change,but it increased comparable with control group;The content of IgM has been enhanced significantly in the 21th day.The content of IgM increased comparable with control group in other days.The content of GOT,GPT,AKP increased significantly in the 14th day,and following the experimental proceeding,it reduced to common gradually.The content of serum calcium and SA have no significant change.
(2)The effects of Prinsepia Utilis Royle Fruit Meal on the immunity of RBC and lymphocyte in chickens as flow:The rate of E-C_(3b)RR and E-ICR have no significantly change comparable with control group,and the rate of ANAE+ stain T lymphocyte、erthrocyte active rosette and erythrocyte total rosette all increased significantly in each age(P＜0.01).
(3)The effects of Prinsepia Utilis Royle Fruit Meal on the immunocompetent cell in immune organ in chickens as flow:Compared with control group,the percent of T lymphocyte in experimental group increased significantly in the early days at 14、21d(P＜0.05 or P＜0.01).The percent of macrophage in experimental group increased significantly in later period at 21、28d (P＜0.05 or P＜0.01).
(4)The effects of Prinsepia Utilis Royle Fruit Meal on the histology of immune organ in chickens as flow:Compared with control group,the weight of thymuses,bursas of Fabricius and spleens of experiment group increased significantly(P＜0.05)or extremely significantly(P＜0.01).Under light microscope,at 14-old-day,the nodulus lymphaticus formed、cortex of thymus is thickening in different lever、spleen Periarterial Lymphatic Sheaths formed and the demarcation between cortex and medullary substance in bursa of test group was more clear than control group;At 21-old-day, the cell differentiation of lymphocytes is maturate than control group,and it has small appearance, and nodulus lymphaticus increased diameter,the number of Hassalls corpuscle more than control group,the thymus cortex broadened.Lymphocytes in spleen align closely;At 28-old-day,the nodulus lymphaticus in bruse and concentric corpuscles of test group was bigger than control group, Langerhans cell have been appear.The germinal center and splenic nodule in test group become prosperous.Under electron microscope,the population of nucleolus increased and nuclear chromation moves around the nuclear membrane and lymphocytes with uniform form in spleen lymphocytes of test group.The activity of lymphocytes increased as time grow.
The results prove that Prinsepia Utilis Royle Fruit Meal has no harmful effects on blood serum biochemical indicator,that it have these function:the level of blood serum antibody could be raised, there was not blight on blood biochemical indicator;there was active regulating action on cellular immunity;it was useful for growth and maturation of immune organ.It is thus evident that Prinsepia Utilis Royle Fruit Meal has rich nutrients which boriler chicks needs.It has Immunomodulation effect for growth of immune organize on boriler chicks in it's early times.
1.青刺果种粕粉营养成分分析本试验首次对青刺果种粕粉进行营养成分的检测,其中,青刺果种粕粉中的氨基酸组分采用氨基酸自动分析仪测定;粗蛋白采用凯氏定氮法;粗脂肪采用索氏抽提法测定;维生素A采用高效液相色谱法;微量元素采用原子吸收光谱法测定。测定结果显示,青刺果种粕粉中各种营养成分的含量如下:氨基酸36.83%;粗蛋白41.26%;粗脂肪6.12%,维生素A 35.40mg╱g,微量元素含量分别为Fe 122.29 ug/g、Zn 54.64 ug/g、Mg 2.47ug/g、Mn 35.01 ug/g、Cu 9.38 ug/g。结果表明青刺果种粕粉中含有丰富的与免疫相关的营养成分。
2.青刺果种粕粉对肉鸡免疫调节作用的研究动物试验:将60只1日龄雏肉鸡随机分为3组,每组20只。Ⅰ组为对照组,饲喂基础日粮;Ⅱ组和Ⅲ组7日龄开始分别在基础日粮中添加10g/kg和15g/kg青刺果种粕粉,将增重最好的组作为试验组。在7、14、21和28日龄采取增重效果最好组采集血样和免疫组织材料(脾脏、法氏囊、胸腺)。血样用于测定血清抗体IgG(Immunoglobulin G)、IgA(Immunoglobulin A)、IgM(Immunoglobulin M)、谷草转氨酶GOT(Glutamic Oxalacetic Transaminase)、谷丙转氨酶GPT(Glutamic Pyruvic Transaminase)、血清钙、血清白蛋白SA(Serum Aibumin)、血清碱性磷酸酶AKP(Alkaline Phosphatase),以及红细胞C3b受体花环率E-C3bRR(Erythrocyte C3b Receptor Ratio)、红细胞免疫复合物花环率E-ICR(Erythro- cyte immune Compound Ratio)、总淋巴细胞玫瑰花环率Et(T-lymphocyteErthrocyte Total Rosette Rates)、活性淋巴细胞玫瑰花环率Ea(T-lymphocyte Erthrocyte ActiveRosette Rates)、T淋巴细胞百分率。免疫组织材料用于测定脏器指数、免疫器官中T淋巴细胞和巨噬细胞数量,同时制作石蜡切片,苏木素伊红HE(Hematoxylin and Eosin Staining)染色后观察免疫器官的组织学结构变化;脾脏组织制作电镜切片,电镜下观察脾脏超微结构的变化。结果显示:
(1)青刺果种粕粉对肉鸡血清抗体及血液生化指标的影响表现为:青刺果种粕粉10g/kg剂量组与对照组相比,IgG在各日龄都表现为显著升高;IgA差异不显著,但与对照组相比均有所提高;IgM在21日龄时差异显著,表现为显著升高,在其它日龄均有所提高。GOT、GPT和AKP在14日龄时,与对照组相比差异显著升高,但随着试验的进程,逐渐降低至正常;SA和钙的含量与对照组相比,差异不显著。
(2)青刺果种粕粉对肉鸡红细胞免疫及T淋巴细胞的影响表现为:青刺果种粕粉10g/kg剂量组与对照组相比,E-C_(3b)RR率和E-ICR率差异不显著;T淋巴细胞ANAE~+率与E-玫瑰花环率在各日龄均较对照组极显著升高(P＜0.01)。
(3)青刺果种粕粉对肉鸡免疫器官中免疫细胞的影响表现为:试验组各免疫器官中的T淋巴细胞数量的含量与对照组相比,主要表现在试验初期(14、21日龄),差异显著(P＜0.05或P＜0.01),表现为不同程度的升高;试验组各免疫器官中的巨噬细胞数量与对照组相比较,主要表现在试验后期(21、28日龄),差异显著,表现为明显升高。
(4)青刺果种粕粉对肉鸡免疫器官发育影响的结果表明,试验组肉鸡法氏囊、胸腺、脾脏脏器指数在各个日龄均显著或极显著高于对照组(P＜0.05或P＜0.01)。光镜下,14日龄时,试验组较对照组法氏囊皮质与髓质分界较清楚,淋巴滤泡形成,胸腺皮质不同程度增厚,脾脏形成动脉周围淋巴鞘;21日龄时,试验组较对照组法氏囊中淋巴细胞发育成熟,形态较小,淋巴滤泡直径增加,胸腺小体增多,皮质增厚,脾脏淋巴细胞致密;28日龄时,试验组与对照组比较,法氏囊淋巴滤泡较大,胸腺小体增大,可见朗罕氏细胞,脾小体和生发中心较发达。电镜下,试验组脾脏淋巴细胞形态均一,核仁增多,核染色质边移,随着日龄的增长,淋巴细胞活性增加。
试验结果证明,青刺果种粕粉对肉鸡的血清生理生化指标无不良影响;能提高血清抗体含量,对的细胞免疫具有积极的调节作用;能促进肉鸡免疫器官的生长发育成熟。由此可见,青刺果种粕粉含有肉鸡生长发育所需要的各种营养物质,对肉鸡的早期生长发育具有免疫调节作用。
The relationship between the immune and nutrition gradually being revealed,Not only for animal nutrition to maintain normal growth and development necessary,but also to maintain the immune system function and the decisive factor is immune activity is fully expressed.Therefore,the nutritional immunology research have gradually become the hot spot.With the nutritional immunology to development gradually,people to turn their attention to the nature of herbaceous plants,Because it provides a balanced,pure nutrition is the best energy to enhance immune function. Through a lot of information that Prinsepia Utilis Royle contains rich nutrients such as water,ash, crude fiber,fat,protein,17 kinds of amino acids,vitamins A,D,E,K,and a variety of microelement. In China,Prinsepia Utilis Royle is rich in resources,it provided a good condition.for research, development and use.But a long time,after Prinsepia Utilis Royle squeezed the Prinsepia Utilis Royle oil,the Prinsepia Utilis Royle Fruit Meal blank in the development and utilization of state.So, study the immunoloregulation effects of Prinsepia Utilis Royle on chicken,by the animal experiment and the nutritional components analysis of Prinsepia Utilis Royle Fruit Meal such as amino acid,crude protein,crude fat,vitamin A and microelement.The main contents are as follows:
1.Analysis of the Nutrition Ingredient of Prinsepia Utilis Royle Fruit Meal Analysis of the Nutrition Ingredient of Prinsepia Utilis Royle Fruit Meal at first time in this experiment.Analysis of the amino acid、crude protein、crude fat、vitamin A and the microelement by the methods of the amino acid autoanalyser、Kjeldah method、Soxhlet、high efficiency liquid chromatography and atomic absorption spectrotype,respectively.The results showed that the Prinsepia Utilis Royle Fruit Meal was a good material with rich nutrients,the content of amino acids is 36.83%,36.83%,the content of crude protein is41.26%,the content of crude fat is 6.12%,the content of vitamin A is 35.40mg/g,the content of microelement Fe、Zn、Mg、Mn、Cu is 122.29 ug/g、54.64 ug/g、2.47 ug/g、35.01 ug/g、9.38 ug/g.The results prove that the Prinsepia Utilis Royle Fruit Meal is rich in nutrition which has immune-related components.;
2.The Immunoloregulative Effectsof Prinsepia Utilis Royle Fruit Meal on Chicken Animal experiment:Sixty 1-day-old chicken were divided into three groups randomly.Group I feeded with basal diet.GroupⅡand groupⅢwas supplemented with 10g/kg and 15g/kg Prinsepia Utilis Royle Fruit Meal in the basal diet respectively.After 14d、21d and 28d the group which effect of weighting is best was picked out to blood and immune organize(thymus,spleen,bursa)sampling.Blood be use to mensurate erythrocyte C3b receptor ratio(E-C3bRR)、erythro- cyte immune compound ratio (E-ICR)、Et、Ea、percent of T lymphocytes、Antibody level(IgG、IgA、IgM)and Blood Biochemical Indicator(Glutamic Oxalacetic Transaminase(GOT),Glutamic Pyruvic Transaminase(GPT),serum calcium,Alkaline Phosphatase(AKP),Seralbumin(SA)).Immune organize be used to mensurate the index of immune organ,the quantity of T lymphocyte and macrophage.And then we detected change of structure after making splices and Hematoxylin and Eosin(HE)staining.Otherwise ultrastructure of spleen was be observed under electron microscope.
The results showed that(1)the effects of Prinsepia Utilis Royle Fruit Meal on the sera antibody and blood biochemical indicator in chickens as flow:Comparable with control group,The content of IgG increased significantly after taken Prinsepia Utilis Royle Fruit Meal.The content of IgA had no significantly change,but it increased comparable with control group;The content of IgM has been enhanced significantly in the 21th day.The content of IgM increased comparable with control group in other days.The content of GOT,GPT,AKP increased significantly in the 14th day,and following the experimental proceeding,it reduced to common gradually.The content of serum calcium and SA have no significant change.
(2)The effects of Prinsepia Utilis Royle Fruit Meal on the immunity of RBC and lymphocyte in chickens as flow:The rate of E-C_(3b)RR and E-ICR have no significantly change comparable with control group,and the rate of ANAE+ stain T lymphocyte、erthrocyte active rosette and erythrocyte total rosette all increased significantly in each age(P＜0.01).
(3)The effects of Prinsepia Utilis Royle Fruit Meal on the immunocompetent cell in immune organ in chickens as flow:Compared with control group,the percent of T lymphocyte in experimental group increased significantly in the early days at 14、21d(P＜0.05 or P＜0.01).The percent of macrophage in experimental group increased significantly in later period at 21、28d (P＜0.05 or P＜0.01).
(4)The effects of Prinsepia Utilis Royle Fruit Meal on the histology of immune organ in chickens as flow:Compared with control group,the weight of thymuses,bursas of Fabricius and spleens of experiment group increased significantly(P＜0.05)or extremely significantly(P＜0.01).Under light microscope,at 14-old-day,the nodulus lymphaticus formed、cortex of thymus is thickening in different lever、spleen Periarterial Lymphatic Sheaths formed and the demarcation between cortex and medullary substance in bursa of test group was more clear than control group;At 21-old-day, the cell differentiation of lymphocytes is maturate than control group,and it has small appearance, and nodulus lymphaticus increased diameter,the number of Hassalls corpuscle more than control group,the thymus cortex broadened.Lymphocytes in spleen align closely;At 28-old-day,the nodulus lymphaticus in bruse and concentric corpuscles of test group was bigger than control group, Langerhans cell have been appear.The germinal center and splenic nodule in test group become prosperous.Under electron microscope,the population of nucleolus increased and nuclear chromation moves around the nuclear membrane and lymphocytes with uniform form in spleen lymphocytes of test group.The activity of lymphocytes increased as time grow.
The results prove that Prinsepia Utilis Royle Fruit Meal has no harmful effects on blood serum biochemical indicator,that it have these function:the level of blood serum antibody could be raised, there was not blight on blood biochemical indicator;there was active regulating action on cellular immunity;it was useful for growth and maturation of immune organ.It is thus evident that Prinsepia Utilis Royle Fruit Meal has rich nutrients which boriler chicks needs.It has Immunomodulation effect for growth of immune organize on boriler chicks in it's early times.
引文
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