非淀粉多糖酶谱优化及对肉鸡排泄物含氮物含量的影响
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摘要
本试验利用胃蛋白酶-胰液素两步酶水解法，分别筛选非淀粉多糖酶---木聚糖酶、β-葡聚糖酶、纤维素酶、果胶酶、β-甘露聚糖酶在玉米-豆粕和小麦-豆粕日粮中的最佳使用剂量，采用五元二次正交旋转试验设计优化5种NSP酶的最佳酶谱。通过AA肉鸡饲养试验，研究所筛选的酶谱对肉鸡生产性能和排泄物中几种含氮成分的影响，初步探讨非淀粉多糖酶对肉鸡舍环境的改善作用。
试验一：探讨5种NSP酶对肉鸡2种日粮离体消化能的调控。选择木聚糖酶、β-葡聚糖酶、纤维素酶、果胶酶、β-甘露聚糖酶，添加剂量0-900μg/g，用胃蛋白酶-胰液素两步酶水解法，测定肉鸡玉米-豆粕、小麦-豆粕日粮的离体消化能，采用二次回归正交旋转组合试验设计，测定NSP酶作用规律和最佳酶谱。结果显示，5种NSP酶对离体消化能的调控效应显示规律性的“S”型曲线，在玉米-豆粕日粮中，五种酶对其离体消化能调控的主次效应为：木聚糖酶＞β-甘露聚糖酶＞β-葡聚糖酶＞纤维素酶＞果胶酶；小麦-豆粕日粮中五种酶对其离体消化能调控效应的主次顺序为木聚糖酶＞果胶酶＞β-葡聚糖酶＞纤维素酶＞β-甘露聚糖酶。木聚糖酶对玉米-豆粕、小麦-豆粕日粮的离体消化能提升程度较高，分别提高0.24MJ/kg和0.40MJ/kg。玉米-豆粕日粮在本试验条件下最佳酶谱为：木聚糖酶34836U/kg、β-葡聚糖酶6762U/kg、纤维素酶1159U/kg、果胶酶873U/kg、β-甘露聚糖酶24536U/kg；小麦-豆粕日粮在本试验条件下最佳酶谱为：木聚糖酶65406U/kg、β-葡聚糖酶10132U/kg、纤维素酶980U/kg、果胶酶502U/kg、β-甘露聚糖酶5141U/kg。
试验二：以肉鸡为试验动物，分别在玉米-豆粕和小麦-豆粕基础日粮中添加试验一筛选的NSP复合酶，测定肉鸡生产性能，评估NSP复合酶的作用效果。同时测定肉鸡排泄物尿酸、尿素含量和氨气产生量，初步探讨NSP酶谱对肉鸡舍环境的影响。试验选用208只1日龄爱拨益加肉公鸡，分成4个处理，每个处理4个重复。处理组分别是：玉米-豆粕组，玉米-豆粕加酶组，小麦-豆粕组和小麦-豆粕加酶组。非淀粉多糖酶的添加剂量采用试验一的筛选剂量。在1-21日龄阶段，小麦-豆粕组肉鸡平均日增重显著低于小麦-豆粕加酶组（P<0.05）；1-21、21-42、1-42日龄小麦-豆粕组肉鸡料重比分别显著（P<0.01,P<0.05,P<0.01）低于小麦-豆粕加酶组；1-21、21-42、1-42日龄玉米-豆粕加酶组料重比比玉米-豆粕组分别降低了6.6%、4.7%、5.3%，但是未达显著水平（P>0.05）。在肉鸡2-6周龄阶段，玉米-豆粕组和玉米-豆粕加酶组中氨气产生量均显著的高于小麦-豆粕组和小麦-豆粕加酶组；在3周龄时，小麦-豆粕加酶组氨气产生量极显著低于小麦-豆粕组（P<0.01）。在肉鸡3-6周龄时，玉米-豆粕加酶组、玉米-豆粕组尿酸含量均极显著高于小麦-豆粕加酶组、小麦-豆粕组（P<0.01）。在2、5周龄时玉米-豆粕加酶组尿素含量显著高于玉米-豆粕组（P<0.05）。
The study was to investigate the effect of non-starch polysaccharides(NSP) enzymes on in vitrodigestive energy(IVDE) of broiler diets by using a two step method with pepsin-pancreatin,then usedquadratic regress-orthogonal rotary design to screen the optimum enzyme combination,then toinvestigate the effects of enzyme combination on production performance and content of nitrogencontaining compounds of broilers to determine the feasibility of enzyme combination.
The first experiment was conducted to discuss the effects of NSP enzymes on the in vitro digestiveenergy(IVDE) of broiler diets of different content of NSP. The first step investigated the dose-responseof the NSP enzymes on the IVDE in diets using one-way randomized experimental design. Five NSPenzymes-xylanase, β-glucanase, cellulose, pectinase, β-mannanase were added to corn-soybean dietand wheat-soybean diet, from0to900μg/g. The second step screened the optimum enzymecombination using quadratic regress-orthogonal rotary design. The method of test IVDE is two-stage invitro digestion assay. The IVDE of broiler diet regulating by NSP enzyme appeared “S” state diagram.The strength of NSP enzyme manipulation of IVDE in corn-soybean diet is:xylanase> β-mannanase>β-glucanase> cellulose> pectase,in wheat-soybean diet the sequence is: xylanase> pectase>β-glucanase> cellulose> β-mannanase. Xylanase had the most strong effect on IVDE on bothcorn-soybean and wheat-soybean diets, improved0.24MJ/kg,0.40MJ/kg, respectively. Incorn-soybean diet the optimum enzyme combination was xylanase34836U/kg, β-glucanase6762U/kg,cellulose1159U/kg, pectase873U/kg, β-mannanase24536U/kg; and in wheat-soybean diet theoptimum enzyme combination was xylanase65406U/kg, β-glucanase10132U/kg, cellulose980U/kg,pectase502U/kg, β-mannanase5141U/kg.
The second experiment was to study the effect of non-starch polysaccharides enzyme onperformance, uric acid, urea and ammonia in manure of broilers.208one-day old Arbor Acres malebroilers were randomly divided into four groups(corn-soybean diet with NSP enzymegroup,corn-soybean diet group,wheat-soybean diet with NSP enzyme group,wheat-soybean dietgroup),each with4replicates. The composition and level of NSP enzyme in corn-soybean diet andwheat-soybean diet were from the first experiment. The results showed:, wheat-soybean diet group hadlower daily weight gain than wheat-soybean diet+E group during1-21days (P<0.05); wheat-soybeandiet+E group had lower F/G than wheat-soybean diet group during1-21(P<0.01)、21-42(P<0.05)、1-42(P<0.01) days respectively; the F/G of corn-soybean diet+E group was lower than corn-soybeandiet group by6.6%、4.7%、5.3%during1-21、21-42、1-42days,respectively (P>0.05). The manureammonia concentration of corn-soybean diet+E group and corn-soybean diet group were significantlyhigher than wheat-soybean diet+E group and wheat-soybean diet group (P<0.01) from2to6week age;the ammonia concentration of corn-soybean diet+E group was significantly higher than wheat-soybean diet group (P<0.01) at3week age. Compared with wheat-soybean diet+E group and wheat-soybean dietgroup, the uric acid content of corn-soybean diet+E group and corn-soybean diet group weresignificantly improved (P<0.01) in3to6week age. Compared with corn-soybean diet group, the ureacontent of corn-soybean diet+E group were significantly improved at2(P<0.05) and5(P<0.01) weekage,respectively.
试验一：探讨5种NSP酶对肉鸡2种日粮离体消化能的调控。选择木聚糖酶、β-葡聚糖酶、纤维素酶、果胶酶、β-甘露聚糖酶，添加剂量0-900μg/g，用胃蛋白酶-胰液素两步酶水解法，测定肉鸡玉米-豆粕、小麦-豆粕日粮的离体消化能，采用二次回归正交旋转组合试验设计，测定NSP酶作用规律和最佳酶谱。结果显示，5种NSP酶对离体消化能的调控效应显示规律性的“S”型曲线，在玉米-豆粕日粮中，五种酶对其离体消化能调控的主次效应为：木聚糖酶＞β-甘露聚糖酶＞β-葡聚糖酶＞纤维素酶＞果胶酶；小麦-豆粕日粮中五种酶对其离体消化能调控效应的主次顺序为木聚糖酶＞果胶酶＞β-葡聚糖酶＞纤维素酶＞β-甘露聚糖酶。木聚糖酶对玉米-豆粕、小麦-豆粕日粮的离体消化能提升程度较高，分别提高0.24MJ/kg和0.40MJ/kg。玉米-豆粕日粮在本试验条件下最佳酶谱为：木聚糖酶34836U/kg、β-葡聚糖酶6762U/kg、纤维素酶1159U/kg、果胶酶873U/kg、β-甘露聚糖酶24536U/kg；小麦-豆粕日粮在本试验条件下最佳酶谱为：木聚糖酶65406U/kg、β-葡聚糖酶10132U/kg、纤维素酶980U/kg、果胶酶502U/kg、β-甘露聚糖酶5141U/kg。
试验二：以肉鸡为试验动物，分别在玉米-豆粕和小麦-豆粕基础日粮中添加试验一筛选的NSP复合酶，测定肉鸡生产性能，评估NSP复合酶的作用效果。同时测定肉鸡排泄物尿酸、尿素含量和氨气产生量，初步探讨NSP酶谱对肉鸡舍环境的影响。试验选用208只1日龄爱拨益加肉公鸡，分成4个处理，每个处理4个重复。处理组分别是：玉米-豆粕组，玉米-豆粕加酶组，小麦-豆粕组和小麦-豆粕加酶组。非淀粉多糖酶的添加剂量采用试验一的筛选剂量。在1-21日龄阶段，小麦-豆粕组肉鸡平均日增重显著低于小麦-豆粕加酶组（P<0.05）；1-21、21-42、1-42日龄小麦-豆粕组肉鸡料重比分别显著（P<0.01,P<0.05,P<0.01）低于小麦-豆粕加酶组；1-21、21-42、1-42日龄玉米-豆粕加酶组料重比比玉米-豆粕组分别降低了6.6%、4.7%、5.3%，但是未达显著水平（P>0.05）。在肉鸡2-6周龄阶段，玉米-豆粕组和玉米-豆粕加酶组中氨气产生量均显著的高于小麦-豆粕组和小麦-豆粕加酶组；在3周龄时，小麦-豆粕加酶组氨气产生量极显著低于小麦-豆粕组（P<0.01）。在肉鸡3-6周龄时，玉米-豆粕加酶组、玉米-豆粕组尿酸含量均极显著高于小麦-豆粕加酶组、小麦-豆粕组（P<0.01）。在2、5周龄时玉米-豆粕加酶组尿素含量显著高于玉米-豆粕组（P<0.05）。
The study was to investigate the effect of non-starch polysaccharides(NSP) enzymes on in vitrodigestive energy(IVDE) of broiler diets by using a two step method with pepsin-pancreatin,then usedquadratic regress-orthogonal rotary design to screen the optimum enzyme combination,then toinvestigate the effects of enzyme combination on production performance and content of nitrogencontaining compounds of broilers to determine the feasibility of enzyme combination.
The first experiment was conducted to discuss the effects of NSP enzymes on the in vitro digestiveenergy(IVDE) of broiler diets of different content of NSP. The first step investigated the dose-responseof the NSP enzymes on the IVDE in diets using one-way randomized experimental design. Five NSPenzymes-xylanase, β-glucanase, cellulose, pectinase, β-mannanase were added to corn-soybean dietand wheat-soybean diet, from0to900μg/g. The second step screened the optimum enzymecombination using quadratic regress-orthogonal rotary design. The method of test IVDE is two-stage invitro digestion assay. The IVDE of broiler diet regulating by NSP enzyme appeared “S” state diagram.The strength of NSP enzyme manipulation of IVDE in corn-soybean diet is:xylanase> β-mannanase>β-glucanase> cellulose> pectase,in wheat-soybean diet the sequence is: xylanase> pectase>β-glucanase> cellulose> β-mannanase. Xylanase had the most strong effect on IVDE on bothcorn-soybean and wheat-soybean diets, improved0.24MJ/kg,0.40MJ/kg, respectively. Incorn-soybean diet the optimum enzyme combination was xylanase34836U/kg, β-glucanase6762U/kg,cellulose1159U/kg, pectase873U/kg, β-mannanase24536U/kg; and in wheat-soybean diet theoptimum enzyme combination was xylanase65406U/kg, β-glucanase10132U/kg, cellulose980U/kg,pectase502U/kg, β-mannanase5141U/kg.
The second experiment was to study the effect of non-starch polysaccharides enzyme onperformance, uric acid, urea and ammonia in manure of broilers.208one-day old Arbor Acres malebroilers were randomly divided into four groups(corn-soybean diet with NSP enzymegroup,corn-soybean diet group,wheat-soybean diet with NSP enzyme group,wheat-soybean dietgroup),each with4replicates. The composition and level of NSP enzyme in corn-soybean diet andwheat-soybean diet were from the first experiment. The results showed:, wheat-soybean diet group hadlower daily weight gain than wheat-soybean diet+E group during1-21days (P<0.05); wheat-soybeandiet+E group had lower F/G than wheat-soybean diet group during1-21(P<0.01)、21-42(P<0.05)、1-42(P<0.01) days respectively; the F/G of corn-soybean diet+E group was lower than corn-soybeandiet group by6.6%、4.7%、5.3%during1-21、21-42、1-42days,respectively (P>0.05). The manureammonia concentration of corn-soybean diet+E group and corn-soybean diet group were significantlyhigher than wheat-soybean diet+E group and wheat-soybean diet group (P<0.01) from2to6week age;the ammonia concentration of corn-soybean diet+E group was significantly higher than wheat-soybean diet group (P<0.01) at3week age. Compared with wheat-soybean diet+E group and wheat-soybean dietgroup, the uric acid content of corn-soybean diet+E group and corn-soybean diet group weresignificantly improved (P<0.01) in3to6week age. Compared with corn-soybean diet group, the ureacontent of corn-soybean diet+E group were significantly improved at2(P<0.05) and5(P<0.01) weekage,respectively.
引文
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