CN103882094A - Method for screening exogenous carbohydrase spectrum of chicken feed - Google Patents
Method for screening exogenous carbohydrase spectrum of chicken feed Download PDFInfo
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Abstract
The invention provides a method for screening an exogenous carbohydrase spectrum of chicken feed. The method comprises the following steps: selecting a target chicken feed; setting the type and the dose level of the added exogenous carbohydrases; in-vitro simulating gastrointestinal digestion of chicken; detecting the in-vitro digestion emergy of the feed; and analyzing the optimal carbohydrase spectrum of the target feed. According to the method, the animal digesting process is simulated in vitro, and the acting effect of the carbohydrases is represented by using the in-vitro emergy, so that the optimal exogenous carbohydrase spectrum of the chicken feed can be quickly obtained.
Description
Technical field
The present invention relates to the screening method of chicken feed external source carbohydrate zymogram, particularly relate to the method for utilizing in vitro digestion method rapid Optimum chicken feed external source carbohydrate zymogram.
Background technology
Carbohydrate is the main part of feed, is also the main energy derive of animal.Along with feed cost continuous rise, people constantly seek cheaper feedstuff raw material (as: corn processed by product) part and substitute traditional corn and bean-dregs feed raw material.But these processing byproducts often contain higher dietary fiber, and these dietary fibers can not be made full use of by animal body (Anguita et al., 2006).At present, conventionally adopt interpolation external source carbohydrase to improve the utilization ratio of dietary fiber, carbohydrase is often taking non-starch polysaccharide enzyme as main, as zytase (Engberg et al., 2006; Ding Xuemei etc., 2009; Selle et al., 2003), cellulase (Nahm et al., 1985; Chen Xiaochun etc., 2005), beta-glucanase (Philip et al., 1995; Yu et al., 1998) etc., and the action effect of carbohydrase and daily ration composition, the activity of enzyme, zymogram compatibility are closely related.But the uncertain factor such as type and compatibility of external source carbohydrase, causes generally the interpolation of exogenous enzyme being had to certain blindness at present.
The method of the optimum zymogram of screening feed mainly contains intracorporal method (in vivo) and in vitro method (in vitro).Intracorporal method carries out animal experiment exactly, usually cost, effort, time-consuming, systematic error and random error influence factor many (Chen Liang etc., 2013).And the advantage such as that in vitro method has is quick, easy, expense is low, but in vitro method is selected at present parameter and condition usually lack theoretical explanation, have certain randomness (Coles et al., 2005).Therefore, choose rational in vitro digestion parameter, the digestive process of simulation chicken feed, for Different Chicken feed external source carbohydrate components characteristic, the carbohydrate zymogram that screening adapts with it, for giving full play to feed potential, improve efficiency of feed utilization and increase breeding production benefit significant.
Summary of the invention
Technology of the present invention is dealt with problems: in order to overcome the blindness of adding exogenous enzyme in current chicken feed, a kind of rapid screening method of chicken feed external source carbohydrate zymogram is provided, by in vitro method simulated animal digestive process, use the external action effect that can be worth sign carbohydrase of chicken feed, the external source carbohydrate zymogram of quick obtaining chicken feed optimum.
Technical solution of the present invention is as follows: the sieve method of chicken feed external source carbohydrate zymogram, comprises that step is as follows:
(1) choose the chicken feed of target, and in described chicken feed, add the external source carbohydrase of multiple known type, the additive capacity of every kind of external source carbohydrase arranges serial concentration gradient;
(2) the enzyme-added chicken Feed Sample of known weight is placed in to triangular flask, and add damping fluid and the digestive ferment liquid of the simulation peptic digestion of known proportioning, mix, wherein the weight percentage ranges of feed, damping fluid and digestive ferment liquid is 2:6:1～2:10:1, and chicken Feed Sample is simulated to the digestive process of stomach under the simulation peptic digestion parameter of setting;
(3) wait simulating after peptic digestion finishes, in triangular flask, add damping fluid and the digestive ferment liquid of the little intestinal digestion of simulation of known proportioning, wherein the weight percentage ranges of damping fluid and digestive ferment liquid is 19:1～21:1, and continues under the simulation small intestine digestion parameter of setting, chicken Feed Sample to be simulated the digestive process of small intestine;
(4) wait simulating after small intestine digestion finishes, slaking residue is dried to constant weight, measuring residue external digestion can be worth, calculate add the chicken feed external digestion of every kind of carbohydrase can value, and according to the regression relation that carbohydrase and external digestion can be worth, show that every kind of carbohydrase can improve the stagnation point that the chicken feed external digestion of target can value;
(5) every kind of carbohydrase is encoded according to described stagnation point, adopt Orthogonal Regression rotary combination design, can be worth as characteristic index the optimum zymogram of exogenous carbohydrase taking external digestion.
In described step (1), in the chicken feed of target, add 3～7 kinds of external source carbohydrases, additive capacity arranges 0～1000 μ g/g series concentration gradient.
The damping fluid of simulation peptic digestion is in described step (2): pH is 2.0, and molar mass is than 90:8～100:11=sodium-chlor: the mixing buffered soln of Repone K; The enzyme liquid of simulation peptic digestion is: 900～1500U/mL stomach en-liquid of pH2.0.
In described step (2), simulation peptic digestion parameter is: 40～42 DEG C of temperature, shaking speed 120～180rpm, digestion time 4～6h.
The damping fluid of the little intestinal digestion of simulation is in described step (3): pH is 6.3～6.6, and molar mass is than 90:18:152～98:22:186=sodium-chlor: Repone K: phosphatic mixing buffered soln; The enzyme liquid of simulating little intestinal digestion is: pH6.3～6.6, unit enzyme activity is than 37:8:386:1～46:12:464:2=trypsinase: Chymotrypsin: amylase: the mixed enzyme solution of lipase.
In described step (3), the parameter of the little intestinal digestion of simulation is: 40～42 DEG C of temperature, shaking speed 120～180rpm, digestion time 15～18h.
When every kind of carbohydrase coding according to described stagnation point in described step (5), according to the kind of the multiple kinds of carbohydrate enzyme adding, the factor of influence of respective amount is set, each factor of influence arranges 5 levels, adopting Orthogonal Regression rotary combination design to analyze external digestion can be worth, and utilize the external digestion can value and carbohydrase additive capacity relation, symbolize optimum carbohydrate zymogram.Orthogonal Regression rotary combination design has to be calculated simply, and can eliminate the dependency between regression coefficient, reaches the object of the optimum zymogram of rapid Optimum.
The present invention's advantage is compared with prior art:
(1) the present invention is taking experimental data in a large amount of animal bodies as foundation, and the in-vitro simulated parameter of choosing is pressed close to the actual physiological situation of animal more, has the effect of good simulation feed digestive process in chicken body;
(2) the present invention has overcome the blindness of exogenous carbohydrase in the past, use external means efficiently to simulate feed at the GI digestive process of chicken, and propose external digestion and can value characterize optimum zymogram, reach the object of rapid Optimum carbohydrate zymogram.
Brief description of the drawings
Fig. 1 is the schema that the present invention realizes.
Embodiment
Introduce in detail the present invention below in conjunction with drawings and the specific embodiments.But following embodiment only limits to explain the present invention, and protection scope of the present invention should comprise the full content of claim, is not limited only to the present embodiment.
Embodiment 1,
As shown in Figure 1, concrete operation method of the present invention can be implemented as follows:
(1) choose Corn-soybean daily ration, and in feed, add respectively four kinds of external source carbohydrases, be zytase, cellulase, beta-glucanase and polygalacturonase, the concentration gradient level of every kind of external source carbohydrase additive capacity is made as respectively 0,100,200,300,400,500,600,700,800,900,1000 μ g/g.
(2) accurately take 1～2g(and pulverized 60 mesh sieves) raising sample (being accurate to 0.0002g), be placed in 100ml tool plug triangular flask, in triangular flask, add damping fluid and the digestive ferment liquid of simulating peptic digestion, wherein the weight percent of feed, simulation stomach damping fluid and simulation peptic digestion enzyme liquid is 2:8:1; The damping fluid of simulation peptic digestion is that pH is 2.0, and molar mass is than 90:8=sodium-chlor: the mixing buffered soln of Repone K; The 1000U/mL stomach en-liquid that simulation peptic digestion enzyme liquid is pH2.0.After carefully mixing, with rubber plug sealing, in 42 DEG C of water bath with thermostatic control shaking tables (150rpm), start timing after putting into 5min, digestion time is 6h.
(3) after the peptic digestion stage finishes, in triangular flask, add damping fluid and the digestive ferment liquid of simulating little intestinal digestion, wherein the weight percent of damping fluid and digestive ferment liquid is 20:1; The damping fluid of simulating little intestinal digestion is that pH is 6.4, and molar mass is than 90:18:152=sodium-chlor: Repone K: phosphoric acid salt mixing buffered soln; The enzyme liquid of simulating little intestinal digestion is: pH6.4, unit enzyme activity is than the trypsinase of 37:8:386:1: Chymotrypsin: amylase: lipase mixed enzyme solution.After carefully mixing, with rubber plug sealing, in 42 DEG C of water bath with thermostatic control shaking tables (150rpm), start timing after putting into 5min, digestion time is 18h.Slaking residue is dried to constant weight, and measure residue can value, calculating the external digestion of daily ration under every kind of carbohydrase effect of different concns can be worth; And according to the regression relation that carbohydrase and external digestion can be worth, show that it is 600 μ g/g that zytase, cellulase, beta-glucanase and polygalacturonase can improve the stagnation point that the external digestion of target daily ration can value.
(4) four kinds of carbohydrases are made as 4 factors of influence, each factor arranges 5 levels, the critical addition of every kind of carbohydrase (600 μ g/g) is encoded to 0, adopt quaternary quadratic regression orthogonal rotational unitized design, enzyme adds level 400, 500, 600, 700, 800 μ g/g use respectively-2,-1, 0, 1, 2 encode (concrete design factor and level are in tables 1), setting up external digestion can value and the quaternary quadratic regression equation of four kinds of carbohydrase additions, its equation is: the external digestion of Corn-soybean daily ration can value=a0+a1x1+a2x2+a3x3+a4x4+a12x1x2+a13x1x3+a14x1x4+a23x2x3+a 24x2x4+a34x3x4+a11x1
2+ a22x2
2+ a33x3
2+ a44x4
2(a, regression equation coefficient, x1, zytase, x2, cellulase, x3, beta-glucanase, x4, polygalacturonase).Use partial derivative method to solve the combination of carbohydrate zymogram and each carbohydrase additive capacity optimum in regression equation.
Table 1
| Encoded radio | Zytase | Cellulase | Beta-glucanase | Polygalacturonase |
| 2 | 400μg/g | 400μg/g | 400μg/g | 400μg/g |
| 1 | 500μg/g | 500μg/g | 500μg/g | 500μg/g |
| 0 | 600μg/g | 600μg/g | 600μg/g | 600μg/g |
| -1 | 700μg/g | 700μg/g | 700μg/g | 700μg/g |
| -2 | 800μg/g | 800μg/g | 800μg/g | 800μg/g |
Embodiment 2,
(1) choose wheat-dregs of beans daily ration, and in feed, add respectively five kinds of external source carbohydrases, be zytase, cellulase, beta-glucanase, 'beta '-mannase and polygalacturonase, the concentration gradient level of every kind of external source carbohydrase additive capacity is made as respectively 0,100,200,300,400,500,600,700,800,900,1000 μ g/g.
(2) accurately take 1～2g(and pulverized 60 mesh sieves) raising sample (being accurate to 0.0002g), be placed in 100ml tool plug triangular flask, in triangular flask, add damping fluid and the digestive ferment liquid of simulating peptic digestion, wherein the weight percent of feed, simulation stomach damping fluid and simulation peptic digestion enzyme liquid is 2:8:1; The damping fluid of simulation peptic digestion is that pH is 2.0, and molar mass is than 90:8=sodium-chlor: the mixing buffered soln of Repone K; The 1000U/mL stomach en-liquid that simulation peptic digestion enzyme liquid is pH2.0.After carefully mixing, with rubber plug sealing, in 42 DEG C of water bath with thermostatic control shaking tables (150rpm), start timing after putting into 5min, digestion time is 6h.
(3) after the peptic digestion stage finishes, in triangular flask, add damping fluid and the digestive ferment liquid of simulating little intestinal digestion, wherein the weight percent of damping fluid and digestive ferment liquid is 20:1; The damping fluid of simulating little intestinal digestion is that pH is 6.4, and molar mass is than 90:18:152=sodium-chlor: Repone K: phosphoric acid salt mixing buffered soln; The enzyme liquid of simulating little intestinal digestion is: pH6.4, unit enzyme activity is than the trypsinase of 37:8:386:1: Chymotrypsin: amylase: lipase mixed enzyme solution.After carefully mixing, with rubber plug sealing, in 42 DEG C of water bath with thermostatic control shaking tables (150rpm), start timing after putting into 5min, digestion time is 18h.Slaking residue is dried to constant weight, and measure residue can value, calculating the external digestion of daily ration under every kind of carbohydrase effect of different concns can be worth; And according to the regression relation that carbohydrase and external digestion can be worth, show that it is 500 μ g/g that zytase, cellulase, beta-glucanase, 'beta '-mannase and polygalacturonase can improve the stagnation point that the external digestion of target daily ration can value.
(4) five kinds of carbohydrases are made as 5 factors of influence, each factor arranges 5 levels, the critical addition of every kind of carbohydrase (500 μ g/g) is encoded to 0, adopt five yuan of quadratic regression orthogonal rotational unitized design, enzyme adds level 300, 400, 500, 600, 700 μ g/g use respectively-2,-1, 0, 1, 2 encode (concrete design factor and level are in tables 2), setting up external digestion can value and five yuan of quadratic regression equations of five kinds of carbohydrase additions, its equation is: the external digestion of wheat-dregs of beans daily ration can value=a0+a1x1+a2x2+a3x3+a4x4+a5x5+a12x1x2+a13x1x3+a14x1x4+a15x 1x5+a23x2x3+a24x2x4+a25x2x5+a34x3x4+a35x3x5+a45x4x5+a11x 1
2+ a22x2
2+ a33x3
2+ a44x4
2+ a55x5
2(a, regression equation coefficient, x1, zytase, x2, cellulase, x3, beta-glucanase, x4, 'beta '-mannase, x5, polygalacturonase).Use partial derivative method to solve the combination of carbohydrate zymogram and each carbohydrase additive capacity optimum in regression equation.
Table 2
| Encoded radio | Zytase | Cellulase | Beta-glucanase | 'beta '-mannase | Polygalacturonase |
| 2 | 300μg/g | 300μg/g | 300μg/g | 300μg/g | 300μg/g |
| 1 | 400μg/g | 400μg/g | 400μg/g | 400μg/g | 400μg/g |
| 0 | 500μg/g | 500μg/g | 500μg/g | 500μg/g | 500μg/g |
| -1 | 600μg/g | 600μg/g | 600μg/g | 600μg/g | 600μg/g |
| -2 | 700μg/g | 700μg/g | 700μg/g | 700μg/g | 700μg/g |
Embodiment 3,
(1) choose wheat-assorted dregs of rice daily ration, and in feed, add respectively six kinds of external source carbohydrases, be zytase, cellulase, beta-glucanase, 'beta '-mannase, arabanase and polygalacturonase, the concentration gradient level of every kind of external source carbohydrase additive capacity is made as respectively 0,100,200,300,400,500,600,700,800,900,1000 μ g/g.
(2) accurately take 1～2g(and pulverized 60 mesh sieves) raising sample (being accurate to 0.0002g), be placed in 100ml tool plug triangular flask, in triangular flask, add damping fluid and the digestive ferment liquid of simulating peptic digestion, wherein the weight percent of feed, simulation stomach damping fluid and simulation peptic digestion enzyme liquid is 2:8:1; The damping fluid of simulation peptic digestion is that pH is 2.0, and molar mass is than 90:8=sodium-chlor: the mixing buffered soln of Repone K; The 1000U/mL stomach en-liquid that simulation peptic digestion enzyme liquid is pH2.0.After carefully mixing, with rubber plug sealing, in 42 DEG C of water bath with thermostatic control shaking tables (150rpm), start timing after putting into 5min, digestion time is 6h.
(3) after the peptic digestion stage finishes, in triangular flask, add damping fluid and the digestive ferment liquid of simulating little intestinal digestion, wherein the weight percent of damping fluid and digestive ferment liquid is 20:1; The damping fluid of simulating little intestinal digestion is that pH is 6.4, and molar mass is than 90:18:152=sodium-chlor: Repone K: phosphoric acid salt mixing buffered soln; The enzyme liquid of simulating little intestinal digestion is: pH6.4, unit enzyme activity is than the trypsinase of 37:8:386:1: Chymotrypsin: amylase: lipase mixed enzyme solution.After carefully mixing, with rubber plug sealing, in 42 DEG C of water bath with thermostatic control shaking tables (150rpm), start timing after putting into 5min, digestion time is 18h.Slaking residue is dried to constant weight, and measure residue can value, calculating the external digestion of daily ration under every kind of carbohydrase effect of different concns can be worth; And according to the regression relation that carbohydrase and external digestion can be worth, show that it is 700 μ g/g that zytase, cellulase, beta-glucanase, 'beta '-mannase, arabanase and polygalacturonase can improve the stagnation point that the external digestion of target daily ration can value.
(4) six kinds of carbohydrases are made as 6 factors of influence, each factor arranges 5 levels, the critical addition of every kind of carbohydrase (700 μ g/g) is encoded to 0, adopt hexa-atomic quadratic regression orthogonal rotational unitized design, enzyme adds level 500, 600, 700, 800, 900 μ g/g use respectively-2,-1, 0, 1, 2 encode, concrete design factor and level are in table 3, setting up external digestion can value and the hexa-atomic quadratic regression equation of six kinds of carbohydrase additions, its equation is: the external digestion of wheat-assorted dregs of rice daily ration can value=a0+a1x1+a2x2+a3x3+a4x4+a5x5+a6x6+a12x1x2+a13x1x3+a14x1x4+a15x1x5+a16x1x6+a23x2x3+a24x2x4+a25x2x5+a26x2x6+a34x3x4+ a35x3x5+a36x3x6+a45x4x5+a46x4x6+a56x5x6+a11x1
2+ a22x2
2+ a33x3
2+ a44x4
2+ a55x5
2+ a66x6
2(a, regression equation coefficient, x1, zytase, x2, cellulase, x3, beta-glucanase, x4, 'beta '-mannase, x5, arabanase, x6, polygalacturonase).Use partial derivative method to solve the combination of carbohydrate zymogram and each carbohydrase additive capacity optimum in regression equation.
Table 3
It should be noted that, according to the various embodiments described above of the present invention, those skilled in the art are the four corners that can realize independent claim of the present invention and appurtenance completely, implementation procedure and method same the various embodiments described above; And non-elaborated part of the present invention belongs to techniques well known.
The above; be only part embodiment of the present invention, but protection scope of the present invention is not limited to this, in the technical scope that any those skilled in the art disclose in the present invention; the variation that can expect easily or replacement, within all should being encompassed in protection scope of the present invention.
Claims (7)
1. the screening method of chicken feed external source carbohydrate zymogram, is characterized in that performing step is as follows:
(1) choose the chicken feed of target, and in described chicken feed, add the external source carbohydrase of multiple known type, the additive capacity of every kind of external source carbohydrase arranges serial concentration gradient;
(2) the enzyme-added chicken Feed Sample of known weight is placed in to triangular flask, and add damping fluid and the digestive ferment liquid of the simulation peptic digestion of known proportioning, mix, wherein the weight percentage ranges of feed, damping fluid and digestive ferment liquid is 2:6:1～2:10:1, and chicken Feed Sample is simulated to the digestive process of stomach under the simulation peptic digestion parameter of setting;
(3) wait simulating after peptic digestion finishes, in triangular flask, add damping fluid and the digestive ferment liquid of the little intestinal digestion of simulation of known proportioning, wherein the weight percentage ranges of damping fluid and digestive ferment liquid is 19:1～21:1, and continues under the simulation small intestine digestion parameter of setting, chicken Feed Sample to be simulated the digestive process of small intestine;
(4) wait simulating after small intestine digestion finishes, slaking residue is dried to constant weight, measuring residue external digestion can be worth, calculate add the chicken feed external digestion of every kind of carbohydrase can value, and according to the regression relation that carbohydrase and external digestion can be worth, show that every kind of carbohydrase can improve the stagnation point that the chicken feed external digestion of target can value;
(5) every kind of carbohydrase is encoded according to described stagnation point, adopt Orthogonal Regression rotary combination design, can be worth as characteristic index the optimum zymogram of exogenous carbohydrase taking external digestion.
2. the screening method of chicken feed external source carbohydrate zymogram according to claim 1, it is characterized in that: in described step (1), in the chicken feed of target, add 3～7 kinds of external source carbohydrases, additive capacity arranges 0～1000 μ g/g series concentration gradient.
3. the screening method of chicken feed external source carbohydrate zymogram according to claim 1, it is characterized in that: the damping fluid of simulation peptic digestion is in described step (2): pH is 2.0, molar mass is than 90:8～100:11=sodium-chlor: the mixing buffered soln of Repone K; The enzyme liquid of simulation peptic digestion is: 900～1500U/mL stomach en-liquid of pH2.0.
4. the screening method of chicken feed external source carbohydrate zymogram according to claim 1, is characterized in that: in described step (2), simulation peptic digestion parameter is: 40～42 DEG C of temperature, shaking speed 120～180rpm, digestion time 4～6h.
5. the screening method of chicken feed external source carbohydrate zymogram according to claim 1, it is characterized in that: the damping fluid of the little intestinal digestion of simulation is in described step (3): pH is 6.3～6.6, molar mass is than 90:18:152～98:22:186=sodium-chlor: Repone K: phosphatic mixing buffered soln; The enzyme liquid of simulating little intestinal digestion is: pH6.3～6.6, unit enzyme activity is than 37:8:386:1～46:12:464:2=trypsinase: Chymotrypsin: amylase: the mixed enzyme solution of lipase.
6. the screening method of chicken feed external source carbohydrate zymogram according to claim 1, is characterized in that: in described step (3), the parameter of the little intestinal digestion of simulation is: 40～42 DEG C of temperature, shaking speed 120～180rpm, digestion time 15～18h.
7. the screening method of chicken feed external source carbohydrate zymogram according to claim 1, it is characterized in that: when every kind of carbohydrase coding according to described stagnation point in described step (5), according to the kind of the multiple kinds of carbohydrate enzyme adding, the factor of influence of respective amount is set, each factor of influence arranges 5 levels, adopting Orthogonal Regression rotary combination design to analyze external digestion can be worth, and utilize the external digestion can value and carbohydrase additive capacity relation, symbolize optimum carbohydrate zymogram.
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| CN105087754A (en) * | 2015-08-06 | 2015-11-25 | 中国农业科学院北京畜牧兽医研究所 | Fast screening method of pig feed non-starch polysaccharide enzyme |
| CN108559769A (en) * | 2018-04-12 | 2018-09-21 | 上海欧耐施生物技术有限公司 | A kind of appraisal procedure of fodder enzyme |
| CN109385461A (en) * | 2018-10-22 | 2019-02-26 | 中国农业科学院北京畜牧兽医研究所 | A kind of duck diet external source zymogram optimization method |
| US20220020451A1 (en) * | 2020-07-20 | 2022-01-20 | South China Agricultural University | Method for evaluating metabolizable energy in goose diet by using a simulative digestion gross energy technique |
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| CN102242180A (en) * | 2011-03-22 | 2011-11-16 | 北京华美源生物科技有限公司 | Optimal enzyme preparation application system (OEATM) |
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| CN102242180A (en) * | 2011-03-22 | 2011-11-16 | 北京华美源生物科技有限公司 | Optimal enzyme preparation application system (OEATM) |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105087754A (en) * | 2015-08-06 | 2015-11-25 | 中国农业科学院北京畜牧兽医研究所 | Fast screening method of pig feed non-starch polysaccharide enzyme |
| CN105087754B (en) * | 2015-08-06 | 2018-05-08 | 中国农业科学院北京畜牧兽医研究所 | The rapid screening method of one boar food non-starch polysaccharide enzyme spectrum |
| CN108559769A (en) * | 2018-04-12 | 2018-09-21 | 上海欧耐施生物技术有限公司 | A kind of appraisal procedure of fodder enzyme |
| CN109385461A (en) * | 2018-10-22 | 2019-02-26 | 中国农业科学院北京畜牧兽医研究所 | A kind of duck diet external source zymogram optimization method |
| US20220020451A1 (en) * | 2020-07-20 | 2022-01-20 | South China Agricultural University | Method for evaluating metabolizable energy in goose diet by using a simulative digestion gross energy technique |
| US11817181B2 (en) * | 2020-07-20 | 2023-11-14 | South China Agricultural University | Method for evaluating metabolizable energy in goose diet by using a simulative digestion gross energy technique |
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