猪常用饲料能量和粗蛋白质消化率仿生评定方法的研究
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摘要
本研究使用不同纤维水平的日粮，获取生长、育肥猪日粮养分消化率和消化能值等生物学基础数据。利用单胃动物仿生消化仪（SDS-ΙΙ），分析仿生法评定饲料养分消化率的影响因素，并测试日粮和饲料原料的体外消化率，探讨仿生法评定常用猪饲料能量和粗蛋白质消化率的可行性。在“胃——小肠”两步仿生法的研究基础上，重点突破猪“大肠消化”这一环节的模拟测定方法，为猪饲料养分生物学效价仿生评定方法的完善奠定基础。
本研究第一部分首先探明日粮纤维水平和试验期对生长、育肥猪日粮养分在小肠、大肠以及全消化道的消化率和流量，为仿生法模拟饲料养分的消化提供生物学参考数据。本试验将24头荷术猪随机分成4个处理，每个处理6头猪，分别饲喂对照日粮和低、中、高纤维日粮（日粮纤维含量分别为：12.3、14.6、16.9和21.4%）。结果表明：随着日粮纤维含量增加，日粮的干物质、碳水化合物和总能在各个肠段的消化率均线性减少（P <0.05），食糜中干物质、碳水化合物和总能的流量则线性增加（P <0.05），回肠pH值和粪样中的乙酸、丙酸、异丁酸和总VFA含量也线性增加（P <0.01）。从第一试验期到第二试验期，日粮碳水化合物的全消化道消化率和后肠发酵均减少（P <0.01），而回肠食糜中丙酸、异丁酸和总VFA含量则增加（P <0.05）。可溶性和不溶性纤维的摄入量有助于阐释饲料养分消化率和流量的主要变异性（P <0.05）。
第二部分探讨仿生法模拟大肠消化参数和相关测试因素的影响，完善饲料养分生物学效价仿生法评定的操作规程。结果表明：玉米、豆粕和小麦麸的还原糖生成率在消化6h达到最大值（P<0.05）。小麦麸在6个消化时间点的还原糖生成率均显著高于玉米和豆粕的测值（P <0.05）。3种浓度的纤维素酶对饲料的还原糖生成率和大肠能量消化率无显著影响（P>0.05）。消化残渣清洗6次的小麦麸，其能量消化率显著高于清洗3次的值（P <0.05）。透析袋使用两种不同前处理方式，玉米、豆粕和小麦麸干物质消化率无显著影响（P>0.05）。脱脂显著提高玉米、豆粕和小麦麸的干物质和粗脂肪的消化率，粗脂肪消化率在90%以上（P <0.05），但脱脂未对脂肪以外的其它养分的消化率产生影响（P>0.05）。蠕动泵泵入缓冲液以注入式所测得的玉米、豆粕和小麦麸的饲料干物质消化率显著低于吸入式（P <0.05），但其测试的平行性较好。
第三部分研究仿生法评定日粮及饲料原料能量和粗蛋白质的消化率，并与动物试验法测值进行相关性分析，探讨仿生法表征日粮及饲料原料能量和粗蛋白质消化率的可行性。结果表明，仿生法可区分不同饲料的消化率，也可分辨出同一饲料在不同模拟消化阶段的消化率值。仿生法评定纤维日粮的干物质（R2=0.99）、有机物（R2=0.97）和能量消化率（R2=0.97）与动物试验法测值相关性极高（P <0.01），且与日粮中粗纤维含量呈显著负相关（P <0.05）。仿生法测定的消化能值和动物试验法的测值相当接近，差值为0.50±0.24MJ/kg（P <0.05）。仿生法评定饲料原料干物质（R2=0.97，P <0.01）、有机物（R2=0.98，P <0.001）、粗蛋白质（R2=0.82，P <0.05）、能量的消化率（R2=0.93，P <0.01）和消化能值（R2=0.93，P <0.01）与动物试验法测值相关性高。仿生法测定的能量和粗蛋白质的消化率的变异系数范围为0.25~0.84%，动物试验法的变异系数范围为1.00~3.25%（P <0.05）。相对于动物试验法，仿生法测试日粮和饲料原料能量和粗蛋白质消化率的精确度更高。
The effect of dietary fiber level on the nutrient digestibility and digestible energy ofgrowing-fattening pigs was determined in the present study. The effects of factors on the in vitronutrient digestibility of feeds were also analyzed using simulative digestion system (SDS-ΙΙ). Thefeasibility of assaying the in vitro digestibility of energy and crude protein in diets and feed ingredientsusing the SDS-ΙΙ was evaluated by comparing the in vitro and in vivo nutrient digestibility. The in vitromethod for simulating nutrient digestion in swine hindgut was focused in the present study to improvean establishment of a quick and accurate system for feed evaluation, which based on the in vitro methodto simulate digestion in the stomach and small intestine.
The objective of Exp.1was to determine the effects of fiber level from alfalfa meal andexperimental period on the intestinal nutrient flow and hindgut fermentation of growing pigs, in order toprovide data for an establishment of a feed evaluation system using the SDS-ΙΙ. Twenty-four pigs wereprepared by T-cannula insertion into the distal ileum and allotted to4treatments. The pigs wereprovided a control diet or a diet in which corn and soybean meal partly were replaced by5,10and20%,respectively, of alfalfa meal to give the graded levels of dietary fiber during two10-d experimentalperiods (12.3,14.6,16.9and21.4%dietary fiber). The apparent ileal digestibility (AID), apparent totaltract digestibility (ATTD), and hindgut fermentation of dry matter (DM), carbohydrates (CHO), andgross energy (GE) decreased (linear, P <0.05) as the dietary fiber level increased. The intestinal flow ofDM, CHO, and GE increased (linear, P <0.05) with raising the level of dietary fiber. The pH in the ilealdigesta and the concentration of acetate, propionate, and total VFA in the feces increased (linear, P <0.01) as the fiber level increased. From experiment period1to period2, the ATTD and hindgutfermentation of CHO reduced (P <0.01), whereas the concentrations of propionate, valerate, and totalVFA in ileal samples increased (P <0.05). A multiple linear regression analysis taking into accountboth the soluble and insoluble fiber intake explained the main variation (P <0.05) in the total tractdigestibility and flow of nutrients.
The objective of Exp.2was to determine the effects of the factors on the in vitro nutrientdigestibility of feeds to provide a method for meaning the in vitro nutrient digestibility of feeds usingthe SDS-ΙΙ. After preliminary consecutive incubations to simulate digestion in stomach and smallintestine, the in vitro generation rate of reducing sugar (RGG) in all feeds reached maximal value as invitro hindgut digestion for6h, the RGG was greater in wheat bran than in corn and soybean meal (P <0.05). The cellulase concentration did not affect the RGG and in vitro energy digestibility (ED)(P>0.05). The ED of wheat bran during wash times of undigested residues for6times was significantlygreater than for3times (P <0.05). No significant differences were revealed on the in vitro dry matterdigestibility (DMD) of feeds using two preliminary methods of dialysis tube (P>0.05). Removing theoil and fat from undigested residues, the DMD of corn, soybean meal and wheat bran were increased by3%, and the in vitro ether extract digestibility of feeds was more than90%(P <0.05), but the in vitro digestibility of other nutrients was not affected by this method (P>0.05). The DMD of feeds wasdifferent in the pushing and pulling method of running direction of peristaltic pump (P<0.05). Althoughthe DMD of feeds was less in the pushing method than in pulling, the results with well accuracy andprecision were obtained in pushing method.
The objective of Exp.3was determined the in vitro nutrient digestibility of feeds compared withthe in vivo value, so as to evaluating the feasibility of assaying the digestibility of nutrients and energyby the bionic method. The in vitro nutrient digestibility varies in different feed and in different segmentsof simulated digestion. The correlation of digestibility of dry matter (R2=0.99), organic matter (R2=0.97) and energy (R2=0.97) of fiber diets using the in vitro and in vivo method was very high (P <0.01). The in vitro nutrient digestibility was negatively correlated with the content of dietary fiber (P <0.05). The absolute differences between the in vitro and in vivo digestible energy was0.50±0.24MJ/kg(P <0.05). The correlation of the digestibility of dry matter (R2=0.97, P <0.01), organic matter (R2=0.98, P <0.001), crude protein (R2=0.82, P <0.05), energy (R2=0.93, P <0.01) and the digestibleenergy (R2=0.93, P <0.01) of feed ingredients using the in vitro and in vivo method was also very high.The coefficient of variation (0.25~0.84%) of the in vitro energy and protein digestibility was less thanthe coefficient of variation (1.00~3.25%) of the in vivo energy and protein digestibility (P <0.05),which indicated that it can more accurately evaluate digestibility of energy and crude protein in swinefeeds using the in vitro method by the SDS than using the in vivo method.
本研究第一部分首先探明日粮纤维水平和试验期对生长、育肥猪日粮养分在小肠、大肠以及全消化道的消化率和流量，为仿生法模拟饲料养分的消化提供生物学参考数据。本试验将24头荷术猪随机分成4个处理，每个处理6头猪，分别饲喂对照日粮和低、中、高纤维日粮（日粮纤维含量分别为：12.3、14.6、16.9和21.4%）。结果表明：随着日粮纤维含量增加，日粮的干物质、碳水化合物和总能在各个肠段的消化率均线性减少（P <0.05），食糜中干物质、碳水化合物和总能的流量则线性增加（P <0.05），回肠pH值和粪样中的乙酸、丙酸、异丁酸和总VFA含量也线性增加（P <0.01）。从第一试验期到第二试验期，日粮碳水化合物的全消化道消化率和后肠发酵均减少（P <0.01），而回肠食糜中丙酸、异丁酸和总VFA含量则增加（P <0.05）。可溶性和不溶性纤维的摄入量有助于阐释饲料养分消化率和流量的主要变异性（P <0.05）。
第二部分探讨仿生法模拟大肠消化参数和相关测试因素的影响，完善饲料养分生物学效价仿生法评定的操作规程。结果表明：玉米、豆粕和小麦麸的还原糖生成率在消化6h达到最大值（P<0.05）。小麦麸在6个消化时间点的还原糖生成率均显著高于玉米和豆粕的测值（P <0.05）。3种浓度的纤维素酶对饲料的还原糖生成率和大肠能量消化率无显著影响（P>0.05）。消化残渣清洗6次的小麦麸，其能量消化率显著高于清洗3次的值（P <0.05）。透析袋使用两种不同前处理方式，玉米、豆粕和小麦麸干物质消化率无显著影响（P>0.05）。脱脂显著提高玉米、豆粕和小麦麸的干物质和粗脂肪的消化率，粗脂肪消化率在90%以上（P <0.05），但脱脂未对脂肪以外的其它养分的消化率产生影响（P>0.05）。蠕动泵泵入缓冲液以注入式所测得的玉米、豆粕和小麦麸的饲料干物质消化率显著低于吸入式（P <0.05），但其测试的平行性较好。
第三部分研究仿生法评定日粮及饲料原料能量和粗蛋白质的消化率，并与动物试验法测值进行相关性分析，探讨仿生法表征日粮及饲料原料能量和粗蛋白质消化率的可行性。结果表明，仿生法可区分不同饲料的消化率，也可分辨出同一饲料在不同模拟消化阶段的消化率值。仿生法评定纤维日粮的干物质（R2=0.99）、有机物（R2=0.97）和能量消化率（R2=0.97）与动物试验法测值相关性极高（P <0.01），且与日粮中粗纤维含量呈显著负相关（P <0.05）。仿生法测定的消化能值和动物试验法的测值相当接近，差值为0.50±0.24MJ/kg（P <0.05）。仿生法评定饲料原料干物质（R2=0.97，P <0.01）、有机物（R2=0.98，P <0.001）、粗蛋白质（R2=0.82，P <0.05）、能量的消化率（R2=0.93，P <0.01）和消化能值（R2=0.93，P <0.01）与动物试验法测值相关性高。仿生法测定的能量和粗蛋白质的消化率的变异系数范围为0.25~0.84%，动物试验法的变异系数范围为1.00~3.25%（P <0.05）。相对于动物试验法，仿生法测试日粮和饲料原料能量和粗蛋白质消化率的精确度更高。
The effect of dietary fiber level on the nutrient digestibility and digestible energy ofgrowing-fattening pigs was determined in the present study. The effects of factors on the in vitronutrient digestibility of feeds were also analyzed using simulative digestion system (SDS-ΙΙ). Thefeasibility of assaying the in vitro digestibility of energy and crude protein in diets and feed ingredientsusing the SDS-ΙΙ was evaluated by comparing the in vitro and in vivo nutrient digestibility. The in vitromethod for simulating nutrient digestion in swine hindgut was focused in the present study to improvean establishment of a quick and accurate system for feed evaluation, which based on the in vitro methodto simulate digestion in the stomach and small intestine.
The objective of Exp.1was to determine the effects of fiber level from alfalfa meal andexperimental period on the intestinal nutrient flow and hindgut fermentation of growing pigs, in order toprovide data for an establishment of a feed evaluation system using the SDS-ΙΙ. Twenty-four pigs wereprepared by T-cannula insertion into the distal ileum and allotted to4treatments. The pigs wereprovided a control diet or a diet in which corn and soybean meal partly were replaced by5,10and20%,respectively, of alfalfa meal to give the graded levels of dietary fiber during two10-d experimentalperiods (12.3,14.6,16.9and21.4%dietary fiber). The apparent ileal digestibility (AID), apparent totaltract digestibility (ATTD), and hindgut fermentation of dry matter (DM), carbohydrates (CHO), andgross energy (GE) decreased (linear, P <0.05) as the dietary fiber level increased. The intestinal flow ofDM, CHO, and GE increased (linear, P <0.05) with raising the level of dietary fiber. The pH in the ilealdigesta and the concentration of acetate, propionate, and total VFA in the feces increased (linear, P <0.01) as the fiber level increased. From experiment period1to period2, the ATTD and hindgutfermentation of CHO reduced (P <0.01), whereas the concentrations of propionate, valerate, and totalVFA in ileal samples increased (P <0.05). A multiple linear regression analysis taking into accountboth the soluble and insoluble fiber intake explained the main variation (P <0.05) in the total tractdigestibility and flow of nutrients.
The objective of Exp.2was to determine the effects of the factors on the in vitro nutrientdigestibility of feeds to provide a method for meaning the in vitro nutrient digestibility of feeds usingthe SDS-ΙΙ. After preliminary consecutive incubations to simulate digestion in stomach and smallintestine, the in vitro generation rate of reducing sugar (RGG) in all feeds reached maximal value as invitro hindgut digestion for6h, the RGG was greater in wheat bran than in corn and soybean meal (P <0.05). The cellulase concentration did not affect the RGG and in vitro energy digestibility (ED)(P>0.05). The ED of wheat bran during wash times of undigested residues for6times was significantlygreater than for3times (P <0.05). No significant differences were revealed on the in vitro dry matterdigestibility (DMD) of feeds using two preliminary methods of dialysis tube (P>0.05). Removing theoil and fat from undigested residues, the DMD of corn, soybean meal and wheat bran were increased by3%, and the in vitro ether extract digestibility of feeds was more than90%(P <0.05), but the in vitro digestibility of other nutrients was not affected by this method (P>0.05). The DMD of feeds wasdifferent in the pushing and pulling method of running direction of peristaltic pump (P<0.05). Althoughthe DMD of feeds was less in the pushing method than in pulling, the results with well accuracy andprecision were obtained in pushing method.
The objective of Exp.3was determined the in vitro nutrient digestibility of feeds compared withthe in vivo value, so as to evaluating the feasibility of assaying the digestibility of nutrients and energyby the bionic method. The in vitro nutrient digestibility varies in different feed and in different segmentsof simulated digestion. The correlation of digestibility of dry matter (R2=0.99), organic matter (R2=0.97) and energy (R2=0.97) of fiber diets using the in vitro and in vivo method was very high (P <0.01). The in vitro nutrient digestibility was negatively correlated with the content of dietary fiber (P <0.05). The absolute differences between the in vitro and in vivo digestible energy was0.50±0.24MJ/kg(P <0.05). The correlation of the digestibility of dry matter (R2=0.97, P <0.01), organic matter (R2=0.98, P <0.001), crude protein (R2=0.82, P <0.05), energy (R2=0.93, P <0.01) and the digestibleenergy (R2=0.93, P <0.01) of feed ingredients using the in vitro and in vivo method was also very high.The coefficient of variation (0.25~0.84%) of the in vitro energy and protein digestibility was less thanthe coefficient of variation (1.00~3.25%) of the in vivo energy and protein digestibility (P <0.05),which indicated that it can more accurately evaluate digestibility of energy and crude protein in swinefeeds using the in vitro method by the SDS than using the in vivo method.
引文
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