不同来源木聚糖酶及其组合对肉鸡肠道黏膜形态与二糖酶活性及其基因表达的影响
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摘要
本试验旨在探讨不同来源木聚糖酶及其组合对黄羽肉鸡肠道黏膜形态、二糖酶活性及其基因表达的影响。试验选取1 560只1日龄黄羽肉公鸡,随机分为4组(每组6个重复,每个重复65只):对照组(Ⅰ组)饲喂低能量低蛋白质的基础饲粮,试验组(Ⅱ、Ⅲ和Ⅳ组)分别饲喂在基础饲粮中添加4 000 U/kg的黑曲霉来源单一木聚糖酶1#、康氏木霉来源单一木聚糖酶2#和组合型木聚糖酶(木聚糖酶1#∶木聚糖酶2#=1∶1)的饲粮,试验期63 d。每个重复分别于21、42和63日龄时取2只肉鸡采集肠道组织,用于肠道黏膜形态、二糖酶活性及二糖酶基因mRNA表达丰度的测定。结果表明:1)Ⅳ组21日龄时空肠隐窝深度显著低于Ⅰ组(P<0.05);Ⅳ组42日龄时十二指肠绒毛高度/隐窝深度(V/C)值极显著大于Ⅰ组(P<0.01),显著大于Ⅱ和Ⅲ组(P<0.05),且Ⅳ组42日龄时空肠V/C值极显著大于Ⅰ、Ⅱ和Ⅲ组(P<0.01)。2)21日龄时,十二指肠和空肠中麦芽糖酶活性以及空肠中蔗糖酶活性表现为Ⅳ组极显著高于Ⅰ、Ⅱ和Ⅲ组(P<0.01);42日龄时,空肠中蔗糖酶活性表现为Ⅳ组极显著高于Ⅰ和Ⅱ组(P<0.01),显著高于Ⅲ组(P<0.05)。3)42日龄时,Ⅳ组空肠中麦芽糖酶-葡萄糖淀粉酶(MGA)mRNA表达丰度极显著高于Ⅰ和Ⅱ组(P<0.01)。由此得出,肉鸡在前(1~21日龄)、中期(22~42日龄)阶段,木聚糖酶的添加能够改善肉鸡肠道黏膜形态,促进二糖酶的分泌及其基因表达,且组合型木聚糖酶的作用效果优于单一木聚糖酶,表现出高效催化的组合效应。
This experiment was conducted to investigate the effects of different sources of xylanases and their combination on intestinal morphology,disaccharidase activities and gene expression of broilers. A total of 1 560 one-day-old healthy yellow-feather male broilers were randomly divided into 4 groups with 6 replicates per group and 65 broilers per replicate. Broilers in control group( group Ⅰ) were fed a low energy and protein wheat-soybean type basal diet. Broilers in experiment groups( groups Ⅱ,Ⅲ and Ⅳ) were fed the diets supplemented with 4 000 U/kg of xylanase 1# from Aspergillus,xylanase 2# from Trichoderma and the combined xylanase( xylanase 1# ∶xylanase 2# = 1∶1) based on the basal diet,respectively. The experiment period was 63 days. Intestinal tissues were harvested from two broilers of each replicate at 21,42 and 63 days of age for the determination of mucosal morphology,mucosal disaccharidase activities and disaccharidase gene mRNA expression abundances. The results showed as follows: 1) the crypt depth of group Ⅳ in jejunum at 21 days of age was significantly lower than that of group Ⅰ( P<0.05),while the villus height/crypt depth( V/C) value of group Ⅳ in duodenum at 42 days of age was significantly greater than that of group Ⅰ( P < 0. 01) and groups Ⅱ and Ⅲ( P<0.05),and V/C value of group Ⅳ in jejunum at 42 days of age was significantly higher than that of groups Ⅰ,Ⅱ and Ⅲ( P<0.01). 2) At 21 days of age,the activity of maltase in duodenum and jejunum and the activity of sucrase in jejunum of group Ⅳ were significantly higher than those of groups Ⅰ,Ⅱand Ⅲ( P<0.01); at 42 days of age,the activity of sucrase in jejunum of group Ⅳ was significantly higher than that of groups Ⅰand Ⅱ( P<0.01) and group Ⅲ( P<0.05). 3) At 42 days of age,the expression abundance of maltase-glucoamylase( MGA) mRNA in jejunum of group Ⅳ was significantly higher than that of groups Ⅰ and Ⅱ( P<0.01). Thus,the supplementation of xylanase can improve intestinal mucosal morphology and promote the secretion of disaccharidases and their gene expression during broilers in the early( 1 to 21 days of age) and middle( 22 to 42 days of age) stages. And the effect of combined xylanase is better than the single xylanase,showing a combination of efficient catalytic effect.
This experiment was conducted to investigate the effects of different sources of xylanases and their combination on intestinal morphology,disaccharidase activities and gene expression of broilers. A total of 1 560 one-day-old healthy yellow-feather male broilers were randomly divided into 4 groups with 6 replicates per group and 65 broilers per replicate. Broilers in control group( group Ⅰ) were fed a low energy and protein wheat-soybean type basal diet. Broilers in experiment groups( groups Ⅱ,Ⅲ and Ⅳ) were fed the diets supplemented with 4 000 U/kg of xylanase 1# from Aspergillus,xylanase 2# from Trichoderma and the combined xylanase( xylanase 1# ∶xylanase 2# = 1∶1) based on the basal diet,respectively. The experiment period was 63 days. Intestinal tissues were harvested from two broilers of each replicate at 21,42 and 63 days of age for the determination of mucosal morphology,mucosal disaccharidase activities and disaccharidase gene mRNA expression abundances. The results showed as follows: 1) the crypt depth of group Ⅳ in jejunum at 21 days of age was significantly lower than that of group Ⅰ( P<0.05),while the villus height/crypt depth( V/C) value of group Ⅳ in duodenum at 42 days of age was significantly greater than that of group Ⅰ( P < 0. 01) and groups Ⅱ and Ⅲ( P<0.05),and V/C value of group Ⅳ in jejunum at 42 days of age was significantly higher than that of groups Ⅰ,Ⅱ and Ⅲ( P<0.01). 2) At 21 days of age,the activity of maltase in duodenum and jejunum and the activity of sucrase in jejunum of group Ⅳ were significantly higher than those of groups Ⅰ,Ⅱand Ⅲ( P<0.01); at 42 days of age,the activity of sucrase in jejunum of group Ⅳ was significantly higher than that of groups Ⅰand Ⅱ( P<0.01) and group Ⅲ( P<0.05). 3) At 42 days of age,the expression abundance of maltase-glucoamylase( MGA) mRNA in jejunum of group Ⅳ was significantly higher than that of groups Ⅰ and Ⅱ( P<0.01). Thus,the supplementation of xylanase can improve intestinal mucosal morphology and promote the secretion of disaccharidases and their gene expression during broilers in the early( 1 to 21 days of age) and middle( 22 to 42 days of age) stages. And the effect of combined xylanase is better than the single xylanase,showing a combination of efficient catalytic effect.
引文
[1]FRIESEN O D,GUENTER W,MARQUARDT R R,et al.The effect of enzym e supplem entation on the apparent m etabolizable energy and nutrient digestibilities of w heat,barley,oats,and rye for the young broiler chick[J].Poultry Science,1992,71(10):1710-1721.
[2]BRENES A,MARQUARDT R R,GUENTER W,et al.Effect of enzym e supplem entation on the nutritional value of raw,autoclaved,and dehulled lupins(Lupinus albus)in chicken diets[J].Poultry Science,1993,72(12):2281-2293.
[3]SIEO C C,ABDULLAH N,TAN W S,et al.Influence ofβ-glucanase-producing Lactobacillus strains on intestinal characteristics and feed passage rate of broiler chickens[J].Poultry Science,2005,84(5):734-741.
[4]UNI Z,NOY Y,SKLAN D.Posthatch development of sm all intestinal function in the poultry[J].Poultry Science,1999,78(2):215-222.
[5]WU Y B,RAVINDRAN V,THOMAS D G,et al.Influence of m ethod of w hole w heat inclusion and xylanase supplem entation on the perform ance,apparent m etabolisable energy,digestive tract m easurem ents and gut m orphology of broilers[J].British Poultry Science,2004,45(3):385-394.
[6]SENKOYLU N,SAMLI H E,AKYUREK H,et al.Effects of w hole w heat w ith or w ithout xylanase supplem entation on perform ance of layers and digestive organ developm ent[J].Italian Journal of Anim al Science,2009,8(2):155-163.
[7]冯定远.饲料工业的技术创新与技术经济[J].饲料工业,2004(11):1-6.
[8]FRANKEL W L,ZHANG W,AFONSO J,et al.Glutam ine enhancem ent of structure and function in transplanted sm all intestine in the rat[J].Journal of Parenteral and Enteral Nutrition,1993,17(1):47-55.
[9]王子旭,佘锐萍,陈越,等.日粮锌硒水平对肉鸡小肠黏膜结构的影响[J].中国兽医科技,2003,33(7):18-21.
[10]贺永惠,王清华,王艳荣,等.小麦非淀粉多糖与木聚酶对大鼠小肠形态结构的影响[J].河南科技学院学报(自然科学版),2010,38(2):56-59.
[11]范成莉.β-葡聚糖、木聚糖复合酶对断奶仔猪生长轴激素的影响及其机理研究[D].博士学位论文.杭州:浙江大学,2008
[12]丁雪梅,张克英.玉米-杂粕型日粮添加木聚糖酶对肉鸡免疫功能、肠道形态和微生物菌群的影响[J].中国畜牧杂志,2010,46(23):26-30.
[13]GHAYOUR-NAJAFABADI P,KHOSRAVINIA H,GHEISARI A,et al.Productive perform ance,nutrient digestibility and intestinal m orphom etry in broiler chickens fed corn or w heat-based diets supplem ented w ith bacterial-or fungal-originated xylanase[J].Italian Journal of Anim al Science,2017:doi:10.1080/1828051X.2017.1328990.
[14]YANG Y,IJI P,KOCHER A,et al.Effects of xylanase on grow th and gut developm ent of broiler chickens given a w heat-based diet[J].Asian-Australasian Journal of Anim al Sciences,2008,21(11):1659-1664.
[15]孙哲,汪儆,雷祖玉,等.小麦可溶性非淀粉多糖对肉仔鸡小肠粘膜二糖酶活性的影响[J].动物营养学报,2003,15(1):26-30.
[16]许梓荣,李卫芬,孙建义.NSP酶对大麦体外消化的影响[J].中国粮油学报,2002,17(2):51-54.
[17]翁梅倩,夏红萍,钱龙华,等.兔肠发育中与糖消化吸收相关基因的表达[J].上海第二医科大学学报,2001,21(6):495-499.
[18]YADGARY L,YAIR R,UNI Z.The chick embryo yolk sac m em brane expresses nutrient transporter and digestive enzym e genes[J].Poultry Science,2011,90(2):410-416.
[19]MOCHIZUKI K,HONMA K,SHIMADA M,et al.The regulation of jejunal induction of the m altase-glucoam ylase gene by a high-starch/low-fat diet in m ice[J].Molecular Nutrition&Food Research,2010,54(10):1445-1451.
[20]冯定远.新型高效饲料组合酶的最新理论研究与应用技术[J].饲料工业,2011,32(4):1-8.
[2]BRENES A,MARQUARDT R R,GUENTER W,et al.Effect of enzym e supplem entation on the nutritional value of raw,autoclaved,and dehulled lupins(Lupinus albus)in chicken diets[J].Poultry Science,1993,72(12):2281-2293.
[3]SIEO C C,ABDULLAH N,TAN W S,et al.Influence ofβ-glucanase-producing Lactobacillus strains on intestinal characteristics and feed passage rate of broiler chickens[J].Poultry Science,2005,84(5):734-741.
[4]UNI Z,NOY Y,SKLAN D.Posthatch development of sm all intestinal function in the poultry[J].Poultry Science,1999,78(2):215-222.
[5]WU Y B,RAVINDRAN V,THOMAS D G,et al.Influence of m ethod of w hole w heat inclusion and xylanase supplem entation on the perform ance,apparent m etabolisable energy,digestive tract m easurem ents and gut m orphology of broilers[J].British Poultry Science,2004,45(3):385-394.
[6]SENKOYLU N,SAMLI H E,AKYUREK H,et al.Effects of w hole w heat w ith or w ithout xylanase supplem entation on perform ance of layers and digestive organ developm ent[J].Italian Journal of Anim al Science,2009,8(2):155-163.
[7]冯定远.饲料工业的技术创新与技术经济[J].饲料工业,2004(11):1-6.
[8]FRANKEL W L,ZHANG W,AFONSO J,et al.Glutam ine enhancem ent of structure and function in transplanted sm all intestine in the rat[J].Journal of Parenteral and Enteral Nutrition,1993,17(1):47-55.
[9]王子旭,佘锐萍,陈越,等.日粮锌硒水平对肉鸡小肠黏膜结构的影响[J].中国兽医科技,2003,33(7):18-21.
[10]贺永惠,王清华,王艳荣,等.小麦非淀粉多糖与木聚酶对大鼠小肠形态结构的影响[J].河南科技学院学报(自然科学版),2010,38(2):56-59.
[11]范成莉.β-葡聚糖、木聚糖复合酶对断奶仔猪生长轴激素的影响及其机理研究[D].博士学位论文.杭州:浙江大学,2008
[12]丁雪梅,张克英.玉米-杂粕型日粮添加木聚糖酶对肉鸡免疫功能、肠道形态和微生物菌群的影响[J].中国畜牧杂志,2010,46(23):26-30.
[13]GHAYOUR-NAJAFABADI P,KHOSRAVINIA H,GHEISARI A,et al.Productive perform ance,nutrient digestibility and intestinal m orphom etry in broiler chickens fed corn or w heat-based diets supplem ented w ith bacterial-or fungal-originated xylanase[J].Italian Journal of Anim al Science,2017:doi:10.1080/1828051X.2017.1328990.
[14]YANG Y,IJI P,KOCHER A,et al.Effects of xylanase on grow th and gut developm ent of broiler chickens given a w heat-based diet[J].Asian-Australasian Journal of Anim al Sciences,2008,21(11):1659-1664.
[15]孙哲,汪儆,雷祖玉,等.小麦可溶性非淀粉多糖对肉仔鸡小肠粘膜二糖酶活性的影响[J].动物营养学报,2003,15(1):26-30.
[16]许梓荣,李卫芬,孙建义.NSP酶对大麦体外消化的影响[J].中国粮油学报,2002,17(2):51-54.
[17]翁梅倩,夏红萍,钱龙华,等.兔肠发育中与糖消化吸收相关基因的表达[J].上海第二医科大学学报,2001,21(6):495-499.
[18]YADGARY L,YAIR R,UNI Z.The chick embryo yolk sac m em brane expresses nutrient transporter and digestive enzym e genes[J].Poultry Science,2011,90(2):410-416.
[19]MOCHIZUKI K,HONMA K,SHIMADA M,et al.The regulation of jejunal induction of the m altase-glucoam ylase gene by a high-starch/low-fat diet in m ice[J].Molecular Nutrition&Food Research,2010,54(10):1445-1451.
[20]冯定远.新型高效饲料组合酶的最新理论研究与应用技术[J].饲料工业,2011,32(4):1-8.