Animal Feed Science and Technology 160 (2010) 6272

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Animal Feed Science and Technology

journal homepage: www.elsevier.com/locate/anifeedsci

The prediction of apparent metabolisable energy content of oil seeds

and oil seed by-products for poultry from its chemical components, in
vitro analysis or near-infrared reectance spectroscopy
B. Losada a , P. Garca-Rebollar b , C. lvarez a , P. Cachaldora a ,
c , J. Mndez a , J.C. De Blas b,
M.A. Ibnez
a
b
c

COREN, Sociedad Cooperativa Galega, 32003 Ourense, Spain

Departamento de Produccin Animal, Universidad Politcnica de Madrid, ETS Ingenieros Agrnomos, 28040 Madrid, Spain
Departamento de Estadstica, Universidad Politcnica de Madrid, ETS Ingenieros Agrnomos, 28040 Madrid, Spain

a r t i c l e

i n f o

Article history:
Received 12 January 2010
Received in revised form 23 June 2010
Accepted 30 June 2010
Keywords:
Cockerels
Metabolisable energy content
Energy utilization
Near-infrared reectance spectroscopy
Prediction models

a b s t r a c t
Regression models including chemical composition, in vitro digestibility and near-infrared
reectance spectroscopy (NIRS) were compared to predict the energy content of several
feed ingredients for poultry. The nitrogen-corrected apparent metabolisable energy content
(AMEn) and its proportion on total gross energy (AMEn/GE) were determined in 52 batches
of six protein concentrates; samples were randomly obtained from a poultry feed manufacturing plant throughout a 3-year period. A preliminary trial was also designed to adapt
in vitro methods for prediction of in vivo energy values for poultry. Mean concentrations of
AMEn of the ingredients studied ranged from 5.29 to 14.7 MJ/kg DM and those of AMEn/GE
from 0.260 to 0.609. The most precise model for prediction of AMEn and AMEn/GE values
2
for the whole data analysed was that based on NIRS equations (Rcv
= 0.952 and 0.926,
respectively). The best single chemical predictor was the concentration of neutral detergent bre (NDF) in each ingredient (R2 = 0.721 and 0.736, respectively). Further inclusion
of ether extract content increased R2 to 0.945 and 0.902 for AMEn and AMEn/GE, respectively. A model including linear and quadratic effects of in vitro organic matter digestibility
(ivOMd) provided a slightly inferior prediction of AMEn/GE values (R2 = 0.865). However,
the prediction of AMEn from ivOMd was clearly lower (R2 = 0.756), as variations among
batches of GE concentration (from 19.0 to 24.9 kcal/kg DM) were poorly related to in vitro
digestibility values. All the indirect methods used were useful to explain part of the variability of the energy value of the ingredients studied, although the best results were generally
obtained from the NIRS equations. Otherwise, the signicance of the prediction depended
directly on the variability of the different data set studied.
2010 Elsevier B.V. All rights reserved.

Abbreviations: ADF, acid detergent bre; ADL, acid detergent lignin; AOAC, Association of Ofcial Analytical Chemists; NIRS, near-infrared reectance
spectroscopy; AME, apparent metabolisable energy; AMEn, nitrogen-corrected AME; DM, dry matter; CP, crude protein; CF, crude bre; EE, ether extract; GE,
gross energy; FFSB, full fat soy bean; ivDMd, in vitro digestibility of dry matter; ivOMd, in vitro digestibility of organic matter; NDF, neutral detergent bre;
PKM, palm-kernel meal; SNVD-D, standard normal variate and detrending; R2 , coefcient of determination of regression; Rc2 , coefcient of determination of
2
, coefcient of determination of cross-validation; RSD, residual standard deviation; RSM, rapeseed meal; RPD, ratio of prediction to deviation;
calibration; Rcv
SBM, soybean meal; SD, standard deviation; SEC, standard error of calibration; SECV, standard error of cross-validation; SFM, sunower meal.
Corresponding author. Tel.: +34 914524850; fax: +34 915499763.
E-mail address: c.deblas@upm.es (J.C. De Blas).
0377-8401/$ see front matter 2010 Elsevier B.V. All rights reserved.
doi:10.1016/j.anifeedsci.2010.06.012

B. Losada et al. / Animal Feed Science and Technology 160 (2010) 6272

63

1. Introduction
Indirect techniques, such as chemical analyses, in vitro methods and near-infrared reectance spectroscopy (NIRS), have
been used in different animal species to estimate energy values of raw materials and complete diets. These methods reduce
cost and time of feed evaluation with respect to in vivo trials. However, their capability to estimate feed energy contents
must be validated with in vivo determined values.
Several equations have been proposed in poultry to estimate the apparent metabolisable energy values (AME) of compound feeds from its chemical composition (e.g. Carpenter and Clegg, 1956; Sibbald et al., 1980; Fisher, 1982; Carr et al.,
1984; EEC, 1986). However, these equations are of limited value when they are extrapolated to predict the energy concentration of single feeds, with a different chemical composition than that of complete diets. Furthermore, prediction equations
for feed ingredients are scarce in poultry and its validity is limited to the conditions in which they were obtained (Dolz and
De Blas, 1992; Francesch, 2001). In addition, this approach is also limited by the time required for the chemical analyses and
its repeatability.
Multiple-enzymatic in vitro methods have been proposed as an alternative to chemical analyses to simulate the digestive
processes and to predict energy values in pigs (Boisen and Fernandez, 1997; Noblet and Jacquelin-Peyraud, 2007) and rabbits
(Ramos et al., 1992; Villamide et al., 2008). A two-step in vitro method was tested in poultry by Valdes and Leeson (1992c).
Using this method, the AME of some ingredients, such as toasted and extruded soybeans, was accurately predicted, whereas
those of others, including soybean meal, were greatly overestimated. Otherwise, Losada et al. (2009) determined in cockerels
that a two-step in vitro technique was as accurate as that based on chemical constituents to predict AMEn/gross energy (GE)
efciency of starchy grains and cereal by-products, but that of AME values was poor.
Previous studies have also shown that NIRS allows estimating successfully the major chemical constituents of feeds and
the digestion efciency in several animal species (Farrell, 1999; Roberts et al., 2004). The NIRS method can also predict the
energy values of complete poultry feeds (Valdes and Leeson, 1992a) and those of starch- and bre-concentrated ingredients
for cockerels (Losada et al., 2009) and broiler chickens (Owens et al., 2009). However, the AME values of several raw materials
were not well predicted from NIRS, neither when using specic equations or when extrapolating equations derived from
complete diets (Valdes and Leeson, 1992d, 1994; Garnsworthy et al., 2000).
The aim of this research was to evaluate the interest of using indirect methods (chemical analysis, in vitro digestion and
NIRS techniques), to take into account the variability of in vivo energy values of main protein concentrates ingredients used
by the poultry feed industry.
2. Materials and methods
2.1. Ingredients
A total of 52 batches of full fat soy bean (Glycine max L.) (n = 9), soybean meal, 440 g/kg crude protein (CP) (n = 7); soybean
meal, 470 g/kg CP (n = 12); sunower (Helianthus annuus L.) meal, 320 g/kg CP (n = 12); rapeseed (Brassica napus L.) meal,
351 g/kg CP (n = 7) and palm-kernel (Elaeis Guineensis L.) meal, 160 g/kg CP (n = 5), were randomly sampled from a poultry
feeds manufacturing plant throughout a 3-year period. Samples were analysed immediately upon collection. The number
of samples and the mean and range of chemical composition within each ingredient are shown in Table 1.
2.2. AME determination
The AME value of the feed ingredients was determined in vivo by using the substitution method. Animals were handled
according to the principles for the care of animals in experimentation published by the Spanish Royal Decree 1201/2005
(BOE, 2005), and experiments were approved by the Ethics Committee of the Research and Development Department of
COREN SCG. Experimental diets were manufactured by substituting with the ingredients studied a 50% of a basal diet. The
chemical and raw material composition of the basal diet is shown in Table 2.
The research was conducted in a series of 12 trials, each trial including the evaluation of four to ve feed ingredients and
the corresponding basal diet. Eight cockerels (Warren) were randomly assigned to each experimental diet. Average weight
of animals throughout the trial was 3348 g (120, SD). Birds were housed in individual metabolic cages (0.3 m 0.5 m) with
wire oors, and kept in an environmentally controlled room. Feed ingredients were ground (in a hammer mill, 6 mm screen),
mixed with the basal diets and given in mash form to birds.
Determination of AME of the experimental diets was made following the European reference method (Bourdillon et
al., 1990). The whole droppings were dried in a forced-draught oven at 80 C to constant weight. After drying, the excreta
samples were ground and then stored in a sealed container at 4 C prior to chemical analysis.
The AME values were calculated using the following formula with appropriate corrections made for differences in dry
matter (DM) content:
AME =

(Feed intake GEdiet ) (Excreta output GEexcreta )

Feed intake

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B. Losada et al. / Animal Feed Science and Technology 160 (2010) 6272

Table 1
Chemical composition (g/kg DM) of the feed ingredients studied.
Ingredienta
FFSB
Mean
SDc
Minimum
Maximum

Nb

Moisture

Ash

Crude protein

Ether extract

Crude bre

NDF

ADF

91.4
7.2
82.0
101

49.7
2.1
47.8
54.0

406
5.8
396
412

207
9.2
196
221

62.6
11.8
52.3
90.0

124
17.5
98.0
154

64.8
7.3
56.3
78.9

114
5.5
103
122

67.8
6.0
50.6
72.4

534
19.3
497
562

19.6
9.3
11.4
40.5

49.7
4.2
41.9
58.5

114
15.3
98.0
154

116
6.8
108
129

65.1
2.2
62.8
68.2

491
18.0
475
530

22.5
6.4
11.4
30.5

72.6
7.9
62.8
83.4

101
10.0
85.0
121

78.0
5.4
70.5
86.8

356
37.5
284
403

21.4
9.3
9.2
36.5

105
4.4
99.6
112

73.9
2.3
71.7
77.2

392
14.0
374
409

90.4
5.5
84.6
99.1

55.3
6.1
47.7
62.4

176
4.7
170
182

ADL

Starch

Sugars

Gross energy
(MJ/kg DM)

2.6
2.0
1.1
7.8

5.1
0.5
4.4
5.6

82.6
2.2
78.7
85.3

24.2
0.16
23.1
24.9

60.6
10.3
47.3
79.9

3.2
1.7
1.1
5.6

5.5
0.7
3.4
5.7

102
10.3
92.4
110

19.8
0.18
19.5
20.0

144
88.1
8.6
11.2
130
69.5
152
101

5.0
3.0
2.2
9.0

5.6
0.6
4.5
6.8

94.5
6.4
81.0
101

19.7
0.16
19.5
19.9

218
39.2
161
290

415
234
86.1 13.0
78.0
35.1
24.9
5.9
317
161
50.9
5.5
521
281
110
21.9

78.4
9.1
58.2
89.4

19.4
0.28
19.0
19.9

35.4
10.8
20.0
51.8

123
15.8
98.3
143

333
169
39.7
25.5
307
149
414
215

1.12
0.1
1.11
1.13

80.5
5.9
72.0
86.8

19.9
0.18
19.7
20.2

77.3
12.7
60.9
92.1

204
15.9
182
220

661
385
126
21.8
56.0
12.8
638
338
112
690
447
144

5.5
0.1
5.5
5.6

16.5
2.8
13.0
19.9

20.2
0.38
19.6
20.6

SBM 470 g/kg CP

Mean
SD
Minimum
Maximum

12

SBM 440 g/kg CP

Mean
SD
Minimum
Maximum

SFM 320 g/kg CP

Mean
SD
Minimum
Maximum

12

RSM 351 g/kg CP

Mean
SD
Minimum
Maximum

PKM 160 g/kg CP

Mean
SD
Minimum
Maximum

60.3
4.0
56.0
66.6

NDF, neutral detergent bre; ADF, acid detergent bre; ADL, acid detergent lignin.
a
FFSB, full fat soy bean; SBM, soybean meal; SFM, sunower meal; RSM, rapeseed meal; PKM, palm-kernel meal.
b
n = number of samples.
c
SD = standard deviation.

Nitrogen-corrected AME (AMEn) was calculated by correction to zero nitrogen retention by simple multiplication with
8.22 kcal/g of nitrogen retained in the body as described by Hill and Anderson (1958).
2.3. Chemical analyses
The proximate composition of feed ingredients, experimental diets and excretas were analysed in duplicated samples
using the procedures of AOAC (2000) for dry matter (930.15), ash (923.03), total sugars (974.06), ether extract (920.39) and
crude bre (978.10). Concentration of neutral detergent bre (NDF), acid detergent bre (ADF) and acid detergent lignin (ADL)
was determined according to the sequential method of Van Soest et al. (1991) using heat stable amylase (A3306, Sigma) and
sodium sulphite, and expressed without residual ash. Starch content was measured following the alpha-amyloglucosidase
method (996.11; AOAC, 2000). Nitrogen was measured by combustion (method 968.06; AOAC, 2000) using a VarioMax
CN (Elementar Analysen System, Hanau, Germany). Gross energy was determined in an adiabatic bomb calorimeter (Parr
Instruments, Moline, IL).
2.4. In vitro technique
A preliminary in vitro trial was done to compare the accuracy of prediction of the in vivo degree of metabolizity of gross
energy (AMEn/GE) from several in vitro methodologies. One batch of full fat soy bean (FFSB), soybean meal (SBM, 440 and
470 g/kg CP), sunower meal (SFM) and rapeseed meal (RSM) were incubated using (i) one single step at pH 2 and 39 C
during 2 h with different concentrations of pepsin (2.5, 10 or 15 mg/ml), (ii) adding a second digestion step with pancreatin
(a mixture of protease, amylase and lipase) at pH 6.8 and 39 C during 4 h, and (iii) including a further digestion step using
microbial carbohydrases (Viscozyme 120 L, 120 FBG/g) at pH 4.8 and 39 C during 18 h, as described by Boisen and Fernandez
(1997).

B. Losada et al. / Animal Feed Science and Technology 160 (2010) 6272

65

Table 2
Ingredient and chemical composition of the basal diet (g/kg as fed
basis).
Ingredients
Corn grain
Soybean meal
Lard
Calcium carbonate
Calcium bicarbonate
Monocalcium phosphate
Sodium chloride
Formic acid
Choline chloride
Vitamin-mineral premixa

859
10
9.5
68
0.6
33.5
9.5
2.7
1.6
5.6

Chemical analysis
Crude protein
Lysineb
Threonineb
Methionineb
Crude bre
Starch
Ether extract
Calciumb
Available phosphorusb

69.1
2.2
2.5
1.4
19.3
550
41.3
30.4
8.0

a
Provide, per kg of diet: vitamin A (trans-retinyl acetate),
12,000 IU; vitamin D3 (cholecalciferol), 3000 IU; vitamin E (allrac-tocopherol-acetate), 18 IU; vitamin K3 (bisulphate menadione
complex), 2 mg; pantothenic acid (d-Ca pantothenate), 10 mg;
nicotinic acid, 40 mg; vitamin B12 (cobalamin), 15 g; d-biotin,
80 g; folic acid, 0.5 mg; Se (Na2 SeO3 ), 0.25 mg; I (KI), 1.9 mg; Cu
(CuSO4 5H2 O), 12 mg; Fe (FeSO4 7H2 O), 60 mg; Mn (MnSO4 H2 O),
100 mg; Zn (ZnO), 80 mg.
b
According to FEDNA (2003).

Routine determination of in vitro digestibility of dry matter (ivDMd) and organic matter (ivOMd) of feed ingredients was
made according to the results of the preliminary trial on series of up 30 duplicated samples. In each of the series, a blank was
included. Samples with about 0.5 g of nely ground material (<1 mm) were weighed to an accuracy of 0.1 mg in 100-ml
conical asks. A small magnetic rod and 25 ml of phosphate buffer (0.1 M, pH 6.0) was added to each ask and sample and
buffer were mixed carefully using a magnetic stirring. To the mixture were then added 10 ml 0.2 M HCl, and pH was adjusted
to pH 2.0 with a 1 M ClH or a 1 M NaOH solution, and 1 ml of a eshly prepared pepsin solution containing 2.5 mg pepsin
(porcine, 2000 FIP-U/g, Merck No. 7190). In addition, 0.5 ml of a chloramphenicol solution (0.5 g chloramphenicol, Sigma No.
C-0378), per 100 ml ethanol were added to prevent bacterial growth. Then, the asks were closed with a rubber stopper
and placed on multipoint stirrers in a thermostatically controlled heating chamber (LAN Techniques, Orcoyen, Spain) at
39 C and the samples were stirred gently for 2 h. The in vitro digestibilities of DM and organic matter were calculated from
the difference between concentrations in the sample and the indigested residue, after corrections for values obtained with
reference samples to correct ivDMd and ivOMd values for differences among the successive series.
2.5. NIRS analysis
Spectra from ground (0.7 mm) undried (around 900 g/kg DM) samples of the ingredients studied were collected using a
model 6500 scanning monochromator (FOSS-NIR System, Silver Spring, MD) equipped with a spinning module and standard
ring cups (3.75 cm diameter). The samples were scanned 32 times to create an average spectrum in the reectance mode
over the range of 4002500 nm at 2 nm intervals. Spectra were recorded using the ISI NIRS 3 software ver. 3.11 (Infrasoft
International, Port Matilda, PA). Each ingredient was measured in two independent subsamples and the average spectrum,
stored as log 1/reectance, was used for chemometric analysis.
2.6. Statistics
Prediction equations of in vivo AMEn values of the ingredients tested were developed from chemical and in vitro analysis
by stepwise regression analysis, using the PROC REG of SAS (1990). The stepwise procedure only introduced variables in the
model when they contributed to a signicant improvement (P<0.05) in the estimation of the dependent variable.
All NIRS chemometrics models were developed using modied partial least square regression (MPLS) within the WinISI
III software ver. 1.5. Only the 11002500 nm wavelength range of the sample spectra was used. Prior to calibration, principal
component analysis was performed to remove outliers with a standardized Mahalanobis distance (H) greater than 3.0 (Shenk
and Westerhaus, 1991) and no samples were marked as outliers. For each parameter tested, none and two scatter correction

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B. Losada et al. / Animal Feed Science and Technology 160 (2010) 6272

Table 3
Average values of in vivo apparent metabolisable energy (AMEn, MJ/kg DM), degree of metabolizity of gross energy (AMEn/GE) and in vitro dry matter
(ivDMd) and organic matter (ivOMd) digestibilities of the ingredients studied.
Ingredienta

FFSB
SBM, 470 g/kg CP
SBM, 440 g/kg CP
SFM, 320 g//kg CP
RSM, 351 g/kg CP
PKM, 160 g/kg CP

AMEn

AMEn/GE
b

ivDMd

ivOMd

Mean

SD

Mean

SD

Mean

SD

Mean

SD

14.7
11.1
10.3
7.22
7.88
5.29

0.56
0.39
0.66
1.86
0.84
0.96

0.609
0.562
0.523
0.372
0.396
0.261

0.019
0.020
0.032
0.094
0.042
0.044

0.720
0.648
0.617
0.480
0.522
0.278

0.016
0.015
0.013
0.046
0.012
0.007

0.722
0.655
0.620
0.485
0.531
0.289

0.016
0.016
0.014
0.042
0.014
0.006

DM, dry matter; GE, gross energy, CP, crude protein.

a
FFSB, full fat soy bean; SBM, soybean meal; SFM, sunower meal; RSM, rapeseed meal; PKM, palm-kernel meal.
b
SD = standard deviation.
Table 4
Stepwise regression analysis for AMEn (kJ/kg DM) using chemical composition traits (g/kg DM) as predictors.
Dataa

nb

Step

Regression equationc

R2

RSD

SBM, 470 g/kg CP

SFM, 320 g/kg CP

12
12

1
1

3300 (2309) + 14.7 (4.31) CP

15,925 (1769) 40.0 (7.99) CF

0.536
0.714

277
1041

0.007
<0.001

All soybean meals

19

1
2

2648 (2138) + 15.8 (4.10) CP

5288 (2284) + 11.5 (4.22) CP 107 (49.4) ADL

0.463
0.585

489
443

0.001
<0.001

All soy products

28

1
2

25,045 (1920) 26.9 (3.95) CP

1280 (2631) + 17.2 (4.94) CP + 31.2 (3.22) EE

0.640
0.920

1192
565

<0.001
<0.001

All data

52

1
2

13,683 (402) 14.5 (1.25) NDF

11,935(227) 12.8 (0.59) NDF + 21.5 (1.55) EE

0.721
0.945

1677
770

<0.001
<0.001

AMEn, apparent energy metabolisable energy; DM, dry matter; CP, crude protein; CF, crude bre; ADL, acid detergent lignin; EE, ether extract; NDF, neutral
detergent bre.
a
SBM, soybean meal; SFM, sunower meal.
b
Number of data used to develop the different models.
c
Values in parentheses are standard errors.

techniques, standard normal variate and detrending and multiple scatter correction (Barnes et al., 1989), were compared
together with either or no rst or second order derivatives, giving a total of 14 spectral models for each predicted parameter.
Cross-validation was used to select the optimal number of partial least squares factors and to avoid overtting (Shenk and
Westerhaus, 1995). Calibration was validated using the cross-validation procedure, in which prediction error is evaluated
by splitting the total set of samples into subsets with one subset reserved for validation and the remaining subsets used
for calibration. For each of the quantitative models developed, six subsets for the complete set of samples (n = 52) and eight
subsets for the soybean products (n = 28), soybean meals (n = 19) and sunower meals (n = 12) were applied. No outlier
elimination pass was accepted. Cross-validation was repeated until all subsets were used for validation once, and validation
errors were combined into a standard error of cross-validation (SECV). The statistics used to select the best equations were
2 ) and the SECV. The R2 corresponded to the proportion of reference
the coefcient of determination of cross-validation (Rcv
cv
method variation explained by the cross-validation values and the SECV represent the measurement of the uncertainty. The
ratio of the standard deviation (SD) of the original data to the SECV (ratio of prediction to deviation, RPD, Williams, 2001)
was also used to evaluate calibration performance. The repeatability of the prediction from the NIRS method was estimated
from the variability of the energy values predicted in homogeneous analytical conditions.
3. Results
3.1. Average and standard deviation of AMEn values of the feedstuffs studied
Values of AMEn of the ingredients studied determined by difference are shown in Table 3. Mean values varied from
5.29 (palm-kernel meal, PKM) to 14.7 MJ/kg DM (FFSB). Standard deviations of AMEn and AMEn/GE were, respectively,
3.14 MJ/kg DM and 0.125 for the whole data set of samples studied, and 0.88 MJ/kg DM and 0.042 for the average variation
within ingredients.
3.2. Prediction of AMEn and AMEn/GE from chemical composition
A stepwise regression analysis was made to predict the AMEn of all the ingredients studied from the determined chemical
composition values. The regression equations obtained that reached signicant levels (P<0.05) are presented in Table 4. A
positive effect of the CP concentration in SBM-470 samples (+14.7 kJ/kg DM per each 1 g/kg of increment of CP; n = 12) was

B. Losada et al. / Animal Feed Science and Technology 160 (2010) 6272

67

Fig. 1. Relationship between nitrogen-corrected apparent energy metabolisable (AMEn) and neutral detergent bre (NDF) content of the samples studied.

observed. A similar response (+15.8 kJ) was observed when predicting the AMEn of all the SBM samples (n = 19). In this
instance, the stepwise regression analysis included in a second step a negative effect of ADL content. Inclusion of ADL in
the model allowed the residual standard deviation (RSD) of the equation to decrease (P=0.045) from 0.49 to 0.44 MJ/kg DM,
with respect to an initial standard deviation of all the soybean meal samples of 0.65 MJ/kg DM. When the prediction of
AMEn was made for all the soy products studied, i.e. all the SBM and FFSB samples, an additional effect (P<0.001) of ether
extract (EE) content on AMEn (+31.2 kJ/kg per each 1 g/kg of increment of EE) was also observed, beyond the inuence of CP
concentration. The independent variable that was better correlated to AMEn values of the 12 SFM studied was crude bre
content (r = 0.845), although prediction from ADL and NDF reached similar coefcients of correlation (0.821 and 0.820,
respectively). When all the ingredients were considered (n = 52), the NDF concentration was the rst independent variable
chosen by the stepwise regression analysis, explaining 72.1% of the total variation of the determined AMEn values. The
negative effect of NDF content on AMEn was consistent throughout the whole interval studied (Fig. 1). The concentration of
EE was included at a second step in the model, which allowed decreasing (P<0.001) the RSD of the regression equation up
to 0.77 MJ AMEn/kg DM (see Table 4). This permitted to correct the underestimation of AMEn values of FFSB samples when
dietary NDF content was the only variable in the model (see Fig. 1).
Another stepwise regression equation was calculated to predict the AMEn of the ingredients including as independent
variable the concentration (g/kg DM) of crude bre (CF) instead of NDF:
AMEn(kJ/kg DM) = 12, 056(418) 29.5(2.6)CF + 19.7(2.63)EE
RSD = 1276; n = 52; R2 = 0.842
The stepwise regression equations for AMEn/GE are shown in Table 5. The coefcients of determination were similar
to those determined for AMEn. An exception occurred in the case of the equation obtained for all the soy products, as the
variability of these samples was lower for AMEn/GE than for AMEn (coefcient of variation = 7.0% vs. 16.2%).

Table 5
Stepwise regression analysis for AMEn/GE using chemical composition traits (g/kg DM) as predictors.
Dataa

nb

Step

Regression equationc

R2

RSD

SBM, 470 g/kg CP

SFM, 320 g/kg CP

12
12

1
1

0.202 (0.127) + 0.00067 (0.00024) CP

0.812 (0.090) 0.0020 (0.00041) CF

0.445
0.713

0.015
0.053

0.018
<0.001

All soybean meals

19

1
2

0.187 (0.109) + 0.00070 (0.00021) CP

0.0318 (0.117) + 0.00048 (0.00022) CP 0.0053 (0.0025) ADL

0.394
0.526

0.025
0.023

0.004
0.002

All soy products

28

1
2
3

0.265
0.626
0.671

0.035
0.025
0.024

0.005
<0.001
<0.001

All data

52

1
2

0.737 (0.056) 0.00035 (0.00011) CP

0.191 (0.12) + 0.00066 (0.00022) CP + 0.00072 (0.00015) EE
0.312 (0.13) + 0.00047 (0.00023) CP + 0.00057 (0.0016)
EE 0.0044 (0.0023) ADL
0.641 (0.012) 0.00063 (0.000036) NDF
0.610 (0.012) 0.00060 (0.000031) NDF + 0.00038 (0.000081) EE

0.858
0.902

0.047
0.040

<0.001
<0.001

AMEn, apparent energy metabolisable energy; GE, gross energy; DM, dry matter; CP, crude protein; CF, crude bre; ADL, acid detergent lignin; EE, ether
extract; NDF, neutral detergent bre.
a
SBM, soybean meal; SFM, sunower meal.
b
Number of data used to develop the different models.
c
Values in parentheses are standard errors.

68

B. Losada et al. / Animal Feed Science and Technology 160 (2010) 6272

Fig. 2. Comparison of the ratio between in vivo degree of metabolizity of gross energy and in vitro OM digestibility values determined after one, two or
three digestion steps (1s, 2s, 3s) and using increasing pepsin concentration (2.5, 10 or 25 mg/ml) .

3.3. Prediction of AMEn and AMEn/GE from in vitro digestibilities

The results of the preliminary in vitro trial are presented in Fig. 2. The in vitro organic matter digestibility (ivOMd)
determined using different procedures were compared with in vivo AMEn/GE proportions obtained in the same samples.
The regression analysis showed that the highest correlation (r = 0.970; P<0.001; n = 5) was obtained when the in vitro digestion
was made in one step, using the lowest pepsin concentration (2.5 mg/ml). The correlation coefcients decreased (up to 0.837
and 0.841) at higher pepsin concentrations (10 or 15 mg/ml) or when a two- or three-step technique was followed (0.810
and 0.827, respectively). As shown in Fig. 2, an excess of in vitro digestion led to higher and more homogenous digestibility
estimates than in vivo determinations in this type of samples. According to these results, routine determination of in vitro
digestibility for the remaining samples was done by using the one step procedure with the 2.5 mg/ml pepsin concentration.
The results obtained for each of the ingredients studied are presented in Table 3.
Average values and standard deviation of the in vitro DM and OM digestibility did not differ signicantly, although ivOMd
was slightly higher than ivDMd in all the feedstuffs studied (by 1.43% as average). Values determined of ivOMd ranged from
0.289 (PKM) to 0.722 (FFSB). The standard deviation averaged 0.018, with the highest value corresponding to SFM (0.042).
The regression analysis showed a signicant (P<0.001) linear effect of ivDMd and ivOMd on in vivo AMEn/GE and AMEn
values (see Table 6 and Fig. 3). A signicant (P<0.01) quadratic effect was also observed, as the differences between in vivo
AMEn/GE values for a xed increment of ivOMd were smaller for the high than for the ingredients with a low bre content.
The accuracy of the regression equations was similar when using ivDMd or ivOMd as the independent variable in the model.

Table 6
Equations for prediction of AMEn (kJ/kg DM) and AMEn/GE from in vitro digestibility of DM (ivDMd) and OM (ivOMd).
Dataa

nb

Regression equationc

R2

RSD

(i) AMEn
SFM, 320 g/kg CP

12

8780 (3351) + 21,477 (4476) ivDMd

9530 (3656) + 22,410 (4878) ivOMd
3213 (5137) + 14,596 (5333) ivDMd
3648 (5652) + 15,072 (5882) ivOMd
15,190 (5585) 38,081 (15,068) ivDMd + 36,119 (9852) ivDMd2
16,486 (5940) 41,921 (15,993) ivOMd + 38,767 (10,387) ivOMd2

0.697
0.679
0.305
0.278
0.754
0.756

1071
1104
556
569
1590
1585

<0.001
<0.001
0.014
0.020
<0.001
<0.001

0.445 (0.166) + 1.10 (0.221) ivDMd

0.485 (0.180) + 1.15 (0.24) ivOMd
0.100 (0.250) + 0.674 (0.259) ivDMd
0.118 (0.274) + 0.693 (0.286) ivOMd
0.466 (0.163) 0.991 (0.442) ivDMd + 1.14 (0.29) ivDMd2
0.499 (0.175) 1.10 (0.471) ivOMd + 1.21 (0.31) ivOMd2

0.711
0.694
0.284
0.257
0.867
0.865

0.053
0.054
0.027
0.027
0.046
0.046

<0.001
<0.001
0.019
0.027
<0.001
<0.001

All SBM

19

All data

52

(ii) AMEn/GE
SFM, 320 g/kg CP

12

All SBM

19

All data

52

AMEn, apparent energy metabolisable energy; GE, gross energy.

a
SBM, soybean meal; SFM, sunower meal.
b
Number of data used to develop the different models.
c
Values in parentheses are standard errors.

B. Losada et al. / Animal Feed Science and Technology 160 (2010) 6272

69

Fig. 3. Relationship between nitrogen-corrected apparent energy metabolisable content (AMEn) and in vitro organic matter digestibility (ivOMd) in the
samples studied.

Table 7
2
, SECV) to predict chemical composition (g/kg)
Coefcients of determination and root mean square errors of calibration (Rc2 , SEC) and cross-validation (Rcv
and in vitro digestibility from near-infrared reectance spectra of the ingredients studied (n = 52).

Dry matter
Ash
Crude protein
Ether extract
Crude bre
NDF
ADF
ADL
Total sugars
Gross energy, kJ/kg DM
ivDMd
ivOMd

Maths treatmenta

Factorsb

SECc

Rc2

SECVd

2
Rcv

RPDe

2,4,4,1
0,0,1,1
1,4,4,1
1,4,4,1
0,0,1,1
2,4,4,1
2,4,4,1
2,4,4,1
2,4,4,1
1,4,4,1
2,4,4,1
2,4,4,1

6
4
5
4
7
6
7
6
7
6
5
4

3.12
4.14
7.73
6.25
7.60
17.1
9.81
4.65
2.01
142
0.0173
0.0196

0.930
0.816
0.993
0.991
0.987
0.989
0.990
0.988
0.991
0.994
0.987
0.983

3.76
4.45
7.98
6.86
9.74
21.3
14.7
6.24
2.93
157
0.0200
0.0193

0.899
0.793
0.992
0.989
0.978
0.984
0.977
0.978
0.981
0.992
0.983
0.983

3.14
2.17
11.5
9.35
6.79
7.83
6.60
6.74
7.19
11.2
7.64
7.70

a
Maths treatment, the rst, second, third and fourth numbers denote the order of the derivative function, the spectral segment length over the derivative
is calculated and the segment lengths over which the function was smoothed.
b
Factors, number of actual terms used to estimate the parameter.
c
Standard error of calibration.
d
Standard error of cross-validation.
e
RPD, ratio of prediction to deviation (SD/SECV).

3.4. Prediction of AMEn and AMEn/GE from NIRS analysis

Calibration and cross-validation statistics of prediction of laboratory analyses and energy value of the ingredients studied
from NIRS analysis are shown in Tables 7 and 8, respectively. The coefcients of determination of calibration for chemical
analyses were high, especially for organic constituents (from 0.987 to 0.993), whereas those obtained for cross-validation
were slightly lower (from 0.977 to 0.992). The accuracy of prediction of GE and in vitro digestibility from NIRS spectra for
2 > 0.983). Coefcients of determination of calibration and cross-validation
the whole data set was also very high (Rc2 and Rcv
for prediction of AMEn and AMEn/GE were high (above 0.926) when considering all the data studied. In the current study
these values decreased in parallel to SD of the AMEn values in the data set analysed, as occurred with the other methods of
prediction compared. The information presented in Table 8 is restricted to ingredients (as SFM) or groups of ingredients (all
SBM and all soy products) where the decrease of SD of the energy values in cross-validation was relevant.
4. Discussion
The AMEn concentrations of the ingredients tested in the current study were similar to the average values assigned to
these ingredients by several international feed tables (NRC, 1994; INRA, 2002; FEDNA, 2003; CVB, 2004), and researchers (San
Juan and Villamide, 2000; Rodrigues et al., 2002). The highest deviation was observed for PKM, for which little information
was available. The value determined was 25% below to that assigned by FEDNA (2003) and CVB (2004) tables.
Crude protein content was the independent chemical variable more related to energy concentration in SBM samples,
because of its quantitative relevance, high digestion efciency and negative correlation (r = 0.62; P=0.004) with NDF content.
The coefcient of determination of the equations derived for SBM was low (R2 < 0.585). This might be explained by the small
variation of the determined AMEn values (SD = 0.66, 0.39 and 0.65 MJ/kg DM for SBM-440 and SBM-470, and all the SBM

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B. Losada et al. / Animal Feed Science and Technology 160 (2010) 6272

Table 8
2
, SECV) to predict AMEn (kJ/kg DM) and AMEn/GE
Coefcients of determination and root mean square errors of calibration (Rc2 , SEC) and cross-validation (Rcv
from near-infrared reectance spectra of the ingredients studied.
Dataa

nb

Maths treatmentc

Factorsd

SECe

Rc2

SECVf

2
Rcv

RPDg

(i) AMEn
SFM, 320 g/kg CP
All SBM
All soy products
All data

12
19
28
52

1,4,4,1
0,0,1,1
0,0,1,1
2,4,4,1

3
2
4
7

456
439
456
423

0.940
0.546
0.946
0.982

736
543
527
694

0.858
0.318
0.929
0.952

2.67
1.20
3.71
4.52

(ii) AMEn/GE
SFM, 320 g/kg CP
All SBM
All soy products
All data

12
19
28
52

1,4,4,1
0,0,1,1
0,0,1,1
2,4,4,1

3
2
3
6

0.854
0.248
0.610
0.926

2.63
1.14
1.60
3.69

0.023
0.022
0.021
0.023

0.942
0.504
0.725
0.967

3.57
2.71
2.49
3.39

AMEn, apparent energy metabolisable energy; GE, gross energy.

a
SBM, soybean meal; SFM, sunower meal.
b
Number of data used to develop the different models.
c
Maths treatment, the rst, second, third and fourth numbers denote the order of the derivative function, the spectral segment length over the derivative
is calculated and the segment lengths over which the function was smoothed.
d
Factors, number of actual terms used to estimate the parameter.
e
Standard error of calibration.
f
Standard error of cross-validation.
g
RPD, ratio of prediction to deviation (SD/SECV).

studied, respectively). However, the equations for SBM-47 and for all the SBM data set were signicant (P<0.01) and led to
RSD values of 0.28 and 0.44 MJ/kg DM, respectively.
When all the soy products were considered, the regression coefcient for CP at the rst step was negative, which reects
the higher AMEn and AMEn/GE values of the relatively less protein concentrated FFSB samples (Fig. 4). The greater energy
values of FFSB samples were related to its high EE content (207 g/kg DM as average), which greatly increased its GE and
AMEn concentrations. Furthermore, AMEn/GE efciency in FFSB samples was higher than expected from their NDF content
(Fig. 1). This might be explained by a high digestion efciency of fat (above the average of other nutrients). Moreover, Mateos
et al. (1982) also reported a slower rate of passage and a better absorption of nutrients in poultry diets containing added fat.
After including EE content of the ingredients in the model, the effect of CP on the energy value was again positive. The nal
stepwise regression equation obtained for all the soy products had a high coefcient of determination (R2 = 0.92 for AMEn)
because of the elevated individual variation within this group of samples (SD = 1.94 MJ/kg DM). However, the RSD of this
equation (0.56 MJ/kg DM) was higher than that obtained with the equations for SBM, and was similar to the SD observed
within the different groups of soy samples (see Table 3).
For the SFM, the high SD of the AMEn content of the samples studied (1.86 MJ/kg DM) justies the use of regression
equations to improve the accuracy of estimation of its energy value. The independent variables selected by the model were
those related to bre content, because of its quantitative importance in this feedstuff. A previous study (Villamide and San
Juan, 1998) has also shown that brous constituents were the chemical components that better explained the variation in
the TMEn values of 10 samples of sunower seed meal.
Neutral detergent bre and EE were the variables that explained more of the variation observed in the energy values of
the whole population of samples studied. Use of CF as the independent variable had an inferior RSD than that obtained at the
second step for NDF (Table 4), but can be useful for feed manufacturers using the Weende instead of the Van Soest method
for bre analysis. These equations were highly signicant. However, its usefulness for prediction should take into account

Fig. 4. Relationship between nitrogen-corrected apparent energy metabolisable (AMEn) and crude protein (CP) content in the soy samples studied.

B. Losada et al. / Animal Feed Science and Technology 160 (2010) 6272

71

that their RSD (0.77 MJ AMEn/kg DM and 0.040 for AMEn/GE) were sometimes higher than those determined in equations
derived for single feedstuffs.
The accuracy of the regression equations using in vitro determinations exclusively was poorer than those obtained using
chemical composition (Tables 4 and 5). The prediction of the AMEn of the ingredients tested from in vitro estimations was
improved (up to a RSD value of 0.81 MJ/kg DM, P<0.001) when the EE content was also included in the model, as in vitro
digestibility was little related to changes in gross energy content in the samples studied. Otherwise, the repeatability of
ivDMd and ivOMd (1.27%) was good and similar to that reported for cereal grains and by-products in cockerels (1.05%;

Losada et al., 2009) or for complete diets in pigs (0.9%, Noblet and Jacquelin-Peyraud, 2007) and rabbits (1.09%, Carabano
et al., 2008). These values were respectively similar or better than those obtained for CP or NDF analyses (1.21 and 3.46%,
respectively). Both chemical and in vitro methods had an inferior repeatability than that estimated with the NIRS method,
which were 0.448 and 0.488% for AMEn/GE and AMEn, respectively.
The coefcients of determination and values of SECV obtained conrm the usefulness of NIRS to predict the chemical
composition (Prez-Marn et al., 2004) and the in vitro digestibility in poultry diets (Valdes and Leeson, 1992b). Prediction
from NIRS spectra for the whole data set studied had a SEC and SECV values of 0.42 and 0.69 MJ AMEn/kg DM, which are lower
than those obtained in prediction models using chemical analyses, in vitro digestibility or a combination of both. The direct
calibration of NIRS against in vivo energy values has the advantage of eliminating errors associated to laboratory techniques.
Furthermore, since the spectra contain information of the entire chemistry of the feeds, prediction of the determined energy
values should be better than that obtained from a limited number of chemical predictors. The SEC and SECV values in NIRS
models were lower than SD of most of the ingredients studied. Others (as FFSB and SBM-470) showed little variation in the
random samples collected, so that prediction of its energy concentration was not improved by any of the methods compared
in the current study. The RPD ratio for AMEn and AMEn/GE was 4.52 and 3.69 when the whole data set was studied (Table 7),
which makes the prediction good, according to Williams and Sobering (1996). This ratio should be ideally at least of three,
unless variance of the reference data is low, as it is the case in the current study for some of the sample populations analysed.
Differences in the variance of data also explain the higher coefcients of determination observed for SFM with respect
to those determined for ingredients with a shorter range of variation. The coefcients of determination obtained for AMEn
prediction were similar to those determined in poultry for a similar range of ingredients variation, as samples of wheat
(R2 = 0.45, Garnsworthy et al., 2000). On the other hand, prediction results in the current study were poorer than those
obtained by Valdes and Leeson (1992a) for complete poultry diets, which might be explained by a higher error of the
determination of AMEn by the substitution method as compared to the direct method.
5. Conclusions
The results of the current study indicate that several methods were useful to predict the variation of AMEn values of
protein concentrates for poultry. Near-infrared reectance techniques showed a better accuracy and higher repeatability
than those obtained from chemical analyses or in vitro digestibility data.
Acknowledgement
Financial support was provided by Xunta de Galicia (Project PGDIT05RAG014E).
References
AOAC, 2000. Ofcial Methods of Analysis, 17th ed. Association of Ofcial Analytical Chemists, Washington, DC.
Barnes, R.J., Dhanoa, M.S., Lister, S.J., 1989. Standard normal variate transformation and de-trending of near infrared diffuse reectance spectra. Appl.
Spectrosc. 43, 772777.
Boletn Ocial del Estado (BOE), 2005. Real Decreto 1201/2005. Sobre proteccin de los animales utilizados para experimentacin y otros nes cientcos.
BOE 252, 3436734391.
Boisen, S., Fernandez, J.A., 1997. Prediction of the total tract digestibility of energy in feedstuffs and pig diets by in vitro analyses. Anim. Feed Sci. Technol.
68, 277286.
Bourdillon, A., Carr, B., Conan, L., Duperray, J., Huyghebaert, G., Leclercq, B., Lessire, M., McNab, J., Wiseman, J., 1990. European reference method for the
in vivo determination of metabolisable energy with adult cockerels: reproducibility, effect of food intake and comparison with individual laboratory
methods. Br. Poult. Sci. 31, 557565.
R., Nicodemus, N., Garca, J., Xiccato, G., Trocino, A., Pascual, J.J., Falcao-E-Cunha, L., Maertens, L., 2008. In vitro analysis, an accurate tool to estmate
Carabano,
dry matter digestibility in rabbits. Intra-and inter-laboratory variability. World Rabbit Sci. 16, 195203.
Carpenter, K.J., Clegg, K.M., 1956. The metabolisable energy of poultry feeding-stuffs in relation to their chemical composition. J. Sci. Food Agric. 7, 4551.
Carr, B., Prevotel, B., Leclercq, B., 1984. Cell wall content as a predictor of metabolisable energy value of poultry feedingstuffs. Br. Poult. Sci. 25, 561572.
CVB, 2004. Veevoedertabel (Livestock Feed Table). Centraal Veevoeder Bureau, Lelystad, The Netherlands.
Dolz, S., De Blas, C., 1992. Metabolisable energy of meat and bone meal from Spanish rendering plants as inuenced by level of substitution and method of
determination. Poult. Sci. 71, 316322.
EEC, 1986. Directive de la Commission du 9 avril 1986 xant la mthode de calcul de la valeur nergtique des aliments composs destins la volaille. J.
Off. Commun. Eur. L 130, 5354.
Farrell, D.J., 1999. In vivo and in vitro techniques for the assessment of the energy content of feed grains for poultry: a review. Aust. J. Agric. Res. 50, 881888.
FEDNA, 2003. In: de Blas, C., Mateos, G.G., Garca-Rebollar, P. (Eds.), Tablas FEDNA de composicin y valor nutritivo de alimentos para la fabricacin de

piensos compuestos, 2nd ed. Fundacin Espanola

para el Desarrollo de la Nutricin Animal, Madrid, Spain.
Fisher, C., 1982. Energy Values of Compound Poultry Feeds. Institute for Grassland & Animal Production, Poultry Division, Roslin, Midlothian, UK (Occasional
Publication No. 2.).
Francesch, M., 2001. Sistemas para la valoracin energtica de los alimentos en aves. Arch. Latin Prod. Anim. 9, 3542.

72

B. Losada et al. / Animal Feed Science and Technology 160 (2010) 6272

Garnsworthy, P.C., Wiseman, J., Fegeros, K., 2000. Prediction of chemical, nutritive and agronomic characteristics of wheat by near infrared spectroscopy.
J. Agric. Sci. 135, 409417.
Hill, F.W., Anderson, D.L., 1958. Comparison of metabolisable energy and productive energy determinations with growing chicks. J. Nutr. 64, 587603.
INRA, 2002. Valeurs nutritives pour les volailles. In: Sauvant, D., Perez, J.M., Tran, G. (Eds.), Tables de composition et de valeur nutritive des matires
premires destines aux animaux dlevage. INRA Editions, Paris Cedex, France.
Losada, B., Garca-Rebollar, P., Cachaldora, P., lvarez, C., Mndez, J., De Blas, J.C., 2009. A comparison of the prediction of apparent metabolisable energy
content of starchy grains and cereal by-products for poultry from its chemical components, in vitro analysis or near-infrared reectance spectroscopy.
Span. J. Agric. Res. 7, 813823.
Mateos, G.G., Sell, J., Eastwood, J.A., 1982. Rate of food passage (transit time) as inuenced by level of supplemental fat. Poult. Sci. 61, 94100.
Noblet, J., Jacquelin-Peyraud, Y., 2007. Prediction of organic matter and energy in the growing pig from an invitro method. Anim. Feed Sci. Technol. 134,
211222.
NRC, 1994. Nutrient Requirements of Poultry, 9th ed. National Academy Press, Washington, DC, USA.
Owens, B., McCann, M.E.E., McCracken, K.J., Park, R.S., 2009. Prediction of wheat chemical and physical characteristics and nutritive value by near-infrared
reectance spectroscopy. Br. Poult. Sci. 50, 103122.
Prez-Marn, M.D., Garrido-Vara, A., Guerrero, J.E., Gmez-Cabrera, A., 2004. Near-infrared reectance spectroscopy (NIRS) for the mandatory labelling of
compound feedingstuffs: chemical composition and open-declaration. Anim. Feed Sci. Technol. 116, 333349.
R., Boisen, S., 1992. An in vitro method for estimating digestibility in rabbits. J. Appl. Rabbit Res. 15, 938946.
Ramos, M., Carabano,
Roberts, C.A., Stuth, J., Flinn, P., 2004. Analysis of forages and feedstuffs. In: Roberts, C.A., Workman, J., Reeves, J.B. (Eds.), Near-Infrared Spectroscopy in
Agriculture. American Society of Agronomy, Inc., Madison, IL, USA, pp. 231267.
Rodrigues, P.B., Rostagno, H.S., Teixeira Albino, L.F., Gomes, P.C., Nunes, R.V., Toledo, R.S., 2002. Valores energticos da soja e subprodutos da soja, determinados com frangos de corte e galos adultos. R. Bras. Zootec. 31, 17711782.
San Juan, L.D., Villamide, M.J., 2000. Nutritional evaluation of sunower seed and products derived from them. Effect of oil extraction. Br. Poultry Sci. 41,
182192.
SAS Institute, 1990. SAS/STAT Users guide. Vols. I and II. Version 6, 4th ed. SAS Institute Inc., Cary, NC.
Shenk, J.S., Westerhaus, M.O., 1991. Population structuring of near infrared spectra and modied partial least squares regression. Crop Sci. 31, 15481555.
Shenk, J.S., Westerhaus, M.O., 1995. Development and Network System Management Manual. NIR Systems Inc., Silver Spring, MD.
Sibbald, I.R., Price, K., Barrete, J.P., 1980. True metabolisable energy values for poultry of commercial diets measured by bioassay and predicted from chemical
data. Poult. Sci. 59, 808811.
Valdes, E.V., Leeson, S., 1992a. Near infrared reectance analysis as a method to measure metabolisable energy in complete poultry feeds. Poult. Sci. 71,
11791187.
Valdes, E.V., Leeson, S., 1992b. The use of near infrared reectance spectroscopy to measure in vitro digestible energy content in poultry feeds. Poult. Sci.
71, 13961399.
Valdes, E.V., Leeson, S., 1992c. Measurement of metabolisable energy in poultry feeds by an in vitro system. Poult. Sci. 71, 14931503.
Valdes, E.V., Leeson, S., 1992d. The use of near infrared reectance spectroscopy to measure metabolisable energy in poultry feed ingredients. Poult. Sci.
71, 15591563.
Valdes, E.V., Leeson, S., 1994. Measurement of metabolisable energy, gross energy, and moisture in feed grade fats by near infrared reectance spectroscopy.
Poult. Sci. 73, 163171.
Van Soest, P.J., Robertson, J.B., Lewis, B.A., 1991. Methods for dietary ber, neutral detergent ber and nonstarch polysaccharides in relation to animal
nutrition. J. Dairy Sci. 74, 35833597.
R., Maertens, L., Pascual, J., Gidenne, T., Facao-E-Cunha, L., Xiccato, G., 2008. Prediction of the nutritional value of European
Villamide, M.J., Carabano,
compound feeds for rabbits by chemical components and in vitro analysis. Anim. Feed Sci. Technol. 150, 283294.
Villamide, M.J., San Juan, L.D., 1998. Effect of chemical composition of sunower seed meal on its true metabolizable energy and amino acid digestibility.
Poult. Sci. 77, 18841892.
Williams, P.C., 2001. Implementation of near-infrared technology. In: Williams, P.C., Norris, K. (Eds.), Near-Infrared Technology in the Agricultural and Food
Industries, 2nd ed. American Association of Cereal Chemists Inc., St. Paul, Minnesota, USA, pp. 145169.
Williams, P.C., Sobering, D., 1996. How do we do it: a brief summary of the methods we use in developing near infrared calibrations. In: Davies, A.M.C.,
Williams, P. (Eds.), Near Infrared Spectroscopy: The Future Waves. NIR Publications, Chichester, West Sussex, UK, pp. 185188.