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Ileal amino acid digestibility assay for the

growing meat chicken--comparison of ileal and
excreta amino acid digestibility in the chicken
I.T. Kadim , P.J. Moughan & V. Ravindran
Published online: 28 Jun 2010.

To cite this article: I.T. Kadim , P.J. Moughan & V. Ravindran (2002) Ileal amino acid digestibility assay for the
growing meat chicken--comparison of ileal and excreta amino acid digestibility in the chicken, British Poultry
Science, 43:4, 588-597, DOI: 10.1080/0007166022000004507

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 British Poultry Science (2002) 44: 588–597

1
2
Ileal amino acid digestibility assay for the growing meat chicken–– 3
comparison of ileal and excreta amino acid digestibility in the chicken 4
5
I.T. KADIM, P.J. MOUGHAN AND V. RAVINDRAN 6
7
Institute of Food, Nutrition and Human Health, Massey University, Palmerston North, New Zealand 8
9
10111
1
2
3
4
Abstract 1. The apparent and true amino acid digestibilities in sorghum, wheat, soyabean meal,
5
meat-and-bone meal, Ž sh meal and blood meal for growing meat chickens were determined using
6
an assay based on the collection of digesta from the terminal ileum and comparison was made with
7
digestibility values determined using an excreta-based assay.
8
2. Five-week-old meat chickens were given maize–soyabean meal basal diet or mixtures of the basal
Downloaded by [University of York] at 14:29 01 September 2013

9
diet and test diets containing the 6 ingredients as the sole source of dietary protein (50:50 on weight
20111
basis). Apparent amino acid digestibility values of assay diets at ileal and excreta levels were calcu-
1
lated using chromic oxide as the indigestible marker. True digestibility values were calculated using
2
endogenous outputs determined by feeding a protein-free diet. Amino acid digestibilities of the
3
ingredients were calculated by difference.
4
3. The site of measurement had no in uence on endogenous amino acid output, the exceptions
5
being aspartic acid and glutamic acid. The output of these two amino acids was higher in the excreta.
6
4. SigniŽ cant differences were found between ileal and excreta-based digestibility of certain amino
7
acids in some ingredients, with excreta values being usually higher than the ileal values, indicating
8
a net catabolism of amino acids in the large intestine. The degree of net amino acid disappearance
9
was found to be variable among amino acids and ingredients. In general, threonine had the lowest
30111
digestibility at the ileal level and, compared with other amino acids, the highest degredation during
1
passage through the hindgut.
2
5. The results showed that digestibility determination based on excreta collection will overestimate
3
the uptake for some amino acids in some feeds. The degree of overestimation was often consider-
4
able, ranging from 8·9% (apparent digestibility of threonine in soyabean meal) to 56% (apparent
5
digestibility of aspartic acid in wheat). It is concluded that digestibility values measured at the ter-
6
minal ileum provide a more reliable measure of amino acid availability than those measured in the
7
excreta.
8
9
40111
INTRODUCTION protein composition is avoided (Whitacre and 1
Tanner, 1989). Secondly, the complication aris- 2
Accurate diet formulation for meat chickens ing from the combined voiding of faeces and 3
requires information on digestible rather than urinary amino acids and nitrogen is overcome. 4
gross amino acid contents of dietary ingredients. Moreover, it appears that amino acids are not 5
Consequently, methods have been developed absorbed in the hindgut of the chicken in 6
with poultry to allow the in vivo determination of nutritionally signiŽ cant quantities (Webb, 7
amino acid digestibility (Papadopoulos, 1985; 1990). 8
Low, 1990). These procedures involve the With the sampling of ileal digesta, the 9
sampling and analysis of either ileal digesta or digestibility of a dietary amino acid is deter- 50111
excreta (total tract) from either intact, can- mined by reference to an indigestible marker 1
nulated or caecectomised birds. The ileal included in the test diet. Digestibility values are 2
digestibility assay has two distinct advantages referred to as either apparent or true. The cal- 3
over that based on excreta analysis. First, the culation of apparent digestibility does not take 4
modifying action of the hindgut micro ora on into account endogenous ileal amino acid com- 5
6
Correspondence to: I.T. Kadim. Present address: Animal & Veterinary Sciences Department, College of Agriculture, Sultan Qaboos University, 7
Muscat, Sultanate of Oman. Fax: +968-513-418. E-mail: isam@squ.edu.om 8
Accepted for publication 1st March 2002. 9
60111
ISSN 0007–1668(print)/ISSN 1466–1799(online)/02/030588–10 © 2002 British Poultry Science Ltd
DOI: 10.1080/0007166022000004507
 AMINO ACID DIGESTIBILITY 589

1 ponents whereas with true values the  ow of period to the cages, during which time they
2 amino acids at the terminal ileum is corrected were fed on a commercial broiler starter diet.
3 for those considered to be of endogenous The birds were 30 d old at the commencement
4 (non-dietary) origin. Traditionally, endogenous of the digestibility assay.
5 amino acid losses in birds have been deter-
6 mined following starvation or the ingestion of
Diets
7 a protein-free diet (Sibbald, 1987).
8 An ileal-based amino acid digestibility assay The basal diet contained maize and soyabean
9 for meat chickens has recently been developed meal as the major ingredients. Six test diets
10111 (Kadim and Moughan, 1997a, b; Yap et al., were formulated to contain sorghum, wheat,
1 1997). The main aim of the study reported soyabean meal, meat-and-bone meal, Ž sh meal
2 herein was to compare results from ileal amino or blood meal as the sole source of dietary
3 acid digestibility with those obtained using an protein, and each test diet was combined with
4 excreta-based assay (McNab, 1994). a basal diet (50:50 on a weight basis) to form
5 the 6 assay diets. The ingredient composition
6 of the basal diet and 6 test diets is given in
7 MATERIALS AND METHODS Table 1. The basal diet and assay diets were sub-
8 jected to cold pelleting (60 to 65+C) using a
Animals and housing
2·3 mm die-ring in a pellet mill (Richard Size
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9
20111 Experimental procedures were approved by the Limited Engineers, Orbit 15, Kingston-upon-
1 Massey University Animal Ethics Committee Hull, England). A protein-free diet was also for-
2 and complied with the New Zealand Code of mulated (Table 1) to allow the determination
3 Practice for the Care and Use of Animals for of endogenous  ows of amino acids. This diet
4 ScientiŽ c Purposes. was fed in mash form. Chromic oxide was
5 One hundred and ninety two (23 d old) included in all diets as an indigestible marker.
6 male and female meat chickens (Ross strain)
7 were distributed at random to 48 suspended Procedure
8 wire cages, so that there were two males and two
females per cage. Alternate cages were left After the acclimatisation period, the birds were
9 given their respective diets ad libitum for 4 d and
vacant to avoid cross contamination of excreta.
30111 were fasted for 24 h. The birds were then
The experimental units (cages) were allocated
1 allowed to consume the respective diets for a
at random to the 8 dietary treatments with 6
2 one hour period (Kadim and Moughan, 1997a).
replicates per treatment.
3 Following this, excreta were collected for 13 h
The birds were housed in an environmen-
4 on a tray placed under each cage, transferred
tally controlled room at an ambient tempera-
5 to a plastic container and frozen (–20+C). The
ture of 21+C under a 23-h light and 1-h dark
6 cycle. The birds underwent a 7-d acclimatisation birds were offered the same diet ad libitum for
7
8
9 Table 1. Composition (g/kg, air dry basis) of the basal diet, test diets containing test ingredients and the protein-free diet used in the
experiment
40111
1 Diet1
2 Ingredient B SBM MBM FM BM SOR W PFD
3
4 Soyabean meal 350·00 460·00 - - - - - -
5 Meat-and-bone meal - - 410·00 – – – – –
Fish meal – – – 320·00 – – – –
6 Blood meal – – – – 250·00 – – –
7 Sorghum – – – – – 928·50 – –
8 Wheat – – – – – – 928·50 –
9 Maize 578·50 – – – – – – –
50111 Soyabean oil 30·00 50·00 20·00 30·00 30·00 30·00 30·00 55·00
Maize starch – 368·50 437·50 488·50 555·50 – – 688·50
1 Sucrose – 80·00 80·00 80·00 80·00 – – 170·00
2 Cellulose – – 40·00 40·00 20·00 – – 45·00
3 Potassium carbonate – – 3·00 – 4·00 – – –
4 Sodium chloride 4·00 4·00 2·00 4·00 3·00 4·00 4·00 4·00
5 Dicalcium phosphate 20·00 20·00 10·00 20·00 20·00 20·00 20·00 20·00
Limestone 10·00 10·00 5·00 10·00 10·00 10·00 10·00 10·00
6 Premix2 4·50 4·50 4·50 4·50 4·50 4·50 4·50 4·50
7 Chromic oxide 3·00 3·00 3·00 3·00 3·00 3·00 3·00 3·00
8 1
B = basal; SBM = soyabean meal diet; MBM = meat-and-bone meal diet; FM = Ž sh meal diet; BM = blood meal diet; SOR = sorghum diet;
9 W = wheat diet; PFD = protein–free diet.
60111 2
Broiler Ž nisher vitamin–mineral premix (Technik Products, Auckland, New Zealand).
 590 I.T. KADIM ET AL.
a further 2 d. On d 3 of the second phase of the
experimental period, the birds were again
fasted for 24 h. The birds were then allowed to
consume their diet for a one hour period. Four
hours after the start of the meal, the birds were
killed by an intra-cardial injection of sodium
pentobarbitone. The 4-h sampling time has
been shown to be optimal for ileal digesta sam-
pling in the broiler chicken (Kadim and
Moughan, 1997a). The use of pentobarbitone
minimises peristaltic movements and mucosal
shedding of the gastrointestinal tract, which
may occur with other methods of killing
(Badawy, 1964). When the birds were com-
pletely immobilised, the body cavity was
opened, the ileum removed and digesta gently
 ushed from the ileum (terminal 15 cm adja-
cent to the ileo-caecal junction) using distilled
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water from a syringe. The ileum was deŽ ned as
that section of the intestine from the location
of Meckel’s diverticulum to a point 3 cm prox-
imal to the ileo-caecal junction. Digesta samples
from individual birds were pooled within a cage
to provide adequate material for chemical
analysis and immediately frozen (–20+C). The
excreta and digesta samples were subsequently
freeze-dried, Ž nely ground and stored at –20+C
whilst awaiting chemical analysis.
Chemical analysis
Amino acids contents of duplicate digesta or
excreta samples and quadruplicate diet samples
were determined using a Waters ion-exchange
HPLC system, utilising post-column ninhydrin
derivatisation and  uorescence detection, fol-
lowing hydrolysis in 6M glass-distilled hydrochlo-
ric acid containing 0·1% phenol for 24 h at 110
+ 2+C in evacuated sealed tubes. Lysozyme was
used as an external standard for the amino acid
analysis. Performic acid oxidation was not
undertaken. However, in our laboratory oxygen
removal is virtually complete and recoveries for
methionine exceeding 95% in a standard pro-
tein are routinely attained. Because performic
acid oxidisation was not used in the study, cys-
teine in the samples was not determined. Values
for glycine in the excreta are not presented
because uric acid is non-quantitatively con-
verted to glycine during acid hydrolysis.
Duplicate determinations of dry matter
(DM) and chromium (Cr) were made on diet,
ileal digesta and excreta samples. Dry matter
was determined according to AOAC (1984) pro-
cedures and Cr determinations were made
using atomic absorption spectrophotometry
following the method of Costigan and Ellis
(1987). Where the differences between dupli-
cates for chromium exceeded 2%, samples were
reanalysed. Known amounts of chromic oxide
were included in each Cr run as external stan- 1
dards to check for the recovery of Cr. 2
3
Data analysis 4
5
The amino acid outputs (related to the inges- 6
tion of 1 g of DM; the units are m g/g DM intake) 7
in the ileal digesta or the excreta were calcu- 8
lated using the following equation: 9
Amino acid output =
10111
Amino acid concentration in digesta or excreta 1
´ 2
Diet Cr concentration
__________________________________ 3
Cr concentration in digesta or excreta. 4
5
The amino acid outputs in birds fed the protein- 6
free diets gave the estimations of endogenous 7
concentrations in the ileal digesta and excreta. 8
Apparent amino acid digestibility of the 9
assay diets was calculated using the following 20111
equation (Kadim and Moughan, 1997a). The 1
units are m g/g DM intake. 2
3
Apparent amino acid digestibility (AID) = 4
Amino acid concentration in diet –
Amino acid output (in ileum or excreta) ´ 100. 5
_________________________________ ________ _ 6
Amino acid concentration in diet
7
8
True amino acid digestibility of the assay diets 9
was calculated using the following equation: 30111
True amino acid digestibility = 1
AID + Endogenous amino acid output
_________________________________ ______´ 100. 2
Amino acid concentration in diet 3
4
The apparent and true digestibility of amino 5
acids in the test ingredients were estimated 6
from those in the assay diets (basal/test diet 7
mixtures), using the difference method as 8
described by Fan and Sauer (1995). 9
The ileal and excreta digestibility data were 40111
analysed using a paired t-test (Snedecor and 1
Cochran, 1982). 2
3
4
RESULTS
5
The crude protein (N ´ 6·25) and amino acid 6
contents of the 6 feed ingredients evaluated are 7
presented in Table 2. The quantities of endoge- 8
nous amino acids determined using the two col- 9
lection methods (m g/g DM intake of the 50111
protein-free diet) are given in Table 3. With the 1
exception of aspartic acid and glutamic acid, the 2
endogenous concentrations were similar and not 3
signiŽ cantly different (P > 0·05) at the two sites. 4
The endogenous outputs of these two amino 5
acids were higher (P < 0·05) in the excreta. 6
The apparent and true amino acid 7
digestibility values determined for the test ingre- 8
dients are shown in Tables 4 to 7. Proportional 9
differences in mean amino acid digestibility 60111
 AMINO ACID DIGESTIBILITY 591

1 Table 2. Dry matter, crude protein and amino acid contents (g/kg air dry basis) in the ingredients evaluated in the study
2 Sorghum Wheat Soyabean Meat-and-bone Fish meal Blood meal
3 meal meal
4
Dry matter 952 944 952 946 924 946
5 Crude protein 116 92 471 490 638 913
6 Threonine 3·1 2·9 18·2 15·4 27·9 49·6
7 Valine 4·6 4·5 20·6 21·8 30·0 75·6
8 Methionine 1·6 1·5 6·0 8·4 18·3 9·5
9 Isoleucine 3·9 3·5 19·8 13·7 23·7 7·9
Leucine 14·7 6·5 35·2 30·2 44·8 100·8
10111 Phenylalanine 4·9 3·9 23·7 14·9 26·1 52·9
1 Tyrosine 3·9 3·0 17·7 12·4 19·9 24·0
2 Histidine 2·5 2·3 13·4 10·1 13·9 52·1
3 Lysine 2·2 3·6 29·9 20·0 40·6 78·1
4 Arginine 4·0 5·1 36·8 29·5 42·3 38·4
Aspartic acid 6·7 4·3 47·7 34·8 57·5 80·7
5 Serine 6·6 4·3 24·3 19·0 28·0 49·6
6 Glutamic acid 26·6 27·7 83·7 58·3 76·4 76·6
7 Proline 6·0 9·7 27·7 32·4 37·9 36·7
8 Glycine 2·6 2·6 29·0 58·6 50·3 37·3
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9 Alanine 10·9 3·3 20·0 40·3 41·7 67·7

20111
1 (whenever statistically signiŽ cant differences Table 3. Mean endogenous amino acid outputs (m g/g dry
2 were observed) between the broiler ileal and matter intake)1 determined at the terminal ileum or the excreta
3 excreta-based assays are shown in Table 8. Mean
in broilers
4 of apparent ileal digestibility of the 15 amino Amino acid Ileal Excreta SEM
5 acids ranged from 58·4% for blood meal to
6 Threonine 730 700 39
89·8% for soyabean meal. The range for overall Valine 530 730 52
7 true ileal amino acid digestibility was 63·2% for Methionine 190 170 34
8 blood meal to 96·5% for sorghum. As could be Isoleucine 440 410 53
9 expected, the correction of ileal and excreta Leucine 520 830 65
30111 amino acid outputs, for the endogenous amino
Phenylalanine 410 520 79
1 Tyrosine 390 400 64
acid contribution, resulted in increases in the Histidine 180 310 66
2 values for true digestibility, but the increases Lysine 420 530 37
3 were marked for the cereal grains (sorghum and Arginine 480 400 53
4 wheat) containing lower concentrations of pro- Aspartic acid 600a 1100b 89
5 tein and amino acids. For most amino acids in
Serine 730 900 61
6 Glutamic acid 740a 1700b 69
sorghum and wheat, correction for the endoge- Proline 500 900 62
7 nous component gave rise to digestibility values Glycine 440 – 55
8 exceeding 100%. Alanine 510 700 67
9 In general, amino acid digestibility coefŽ - 1
Values are means of 6 replicate pens (two male and two female
40111 cients based on excreta concentrations were broilers per pen).
1 numerically higher than those determined at the
a,b
Means in the same row with different superscripts are signiŽ -
2 terminal ileum, but only some of the differences
cantly different (P < 0·05).
3 reached statistical signiŽ cance. For apparent
4 digestibility, of the 15 amino acids compared, sig- determined using an assay based on the collec-
5 niŽ cant (P < 0·05) ileal–excreta differences were tion of ileal digesta (Kadim and Moughan,
6 found for 2 amino acids in sorghum, 12 in wheat, 1997a, b) with estimates of digestibility deter-
7 2 in soyabean meal, 9 in meat-and-bone meal, 1 mined in growing meat chickens over the total
8 in Ž sh meal and 4 in blood meal. For true digestive tract. Both apparent and true
9 digestibility, signiŽ cant (P < 0·05) ileal–excreta digestibility values were determined.
50111 differences were recorded for 3 amino acids in To allow determination of true amino acid
1 sorghum, 11 in wheat, 1 in soyabean meal, 5 in digestibility, estimates of the non-dietary
2 meat-and-bone meal, 1 in Ž sh meal and 5 in (endogenous) amino acid components of the
3 blood meal. It is noteworthy that the digestibil- digesta and excreta are required. These were
4 ity of threonine was consistently in uenced by determined by feeding birds an essentially
5 the site of measurement (Table 8). protein-free diet. Interestingly the non-dietary
6 amino acid outputs were similar, when
7 expressed in units of m g/g dry matter intake,
DISCUSSION
8 regardless of whether they were measured in
9 The main objective of the present study was to the ileal digesta or the excreta. It appears that
60111 compare estimates of amino acid digestibility microbial activity in the hindgut of the chicken
 Downloaded by [University of York] at 14:29 01 September 2013

Table 4. Apparent amino acid digestibility (%)1 for sorghum, wheat and soyabean meal determined in the broiler chicken
Sorghum Wheat Soyabean meal
592
Ileal Excreta SEM Ileal Excreta SEM Ileal Excreta SEM

Threonine 62·2a 70·9b 3·01 48·8a 68·4b 2·11 86·9a 94·6b 2·95
Valine 82·3 8·29 3·40 69·4a 80·7b 3·26 88·2 92·6 1·93
Methionine 91·5 92·3 2·32 85·1 86·7 1·52 94·6 92·0 1·02
Isoleucine 88·0 93·5 2·00 75·3a 93·5b 1·14 92·4 93·0 1·55
Leucine 90·1 92·4 1·40 71·9a 92·3b 2·47 88·4 92·3 1·96
Phenylalanine 89·2 88·0 1·06 75·3a 82·2b 1·01 88·1 86·8 2·05
Tyrosine 84·0 86·1 3·93 64·7a 80·4b 2·03 88·4 92·9 2·09
Histidine 78·2 75·4 3·01 70·0a 86·0b 1·18 89·9 93·0 3·02
Lysine 83·1 88·6 2·21 80·8a 90·3b 1·91 93·6 94·6 2·79
Arginine 93·1 89·2 1·23 69·9a 90·2b 3·45 92·4 94·3 1·95
Aspartic acid 85·2 88·1 2·55 56·3a 87·6b 2·54 89·7 92·2 1·02
Serine 80·8a 89·6b 1·29 71·7a 78·2b 1·98 89·3 95·1 1·95
Glutamic acid 91·1 91·2 0·97 89·8 94·1 0·53 90·3 96·9 1·98
Proline 86·2 86·3 1·05 87·2 91·5 0·95 88·1a 96·1b 2·05
Glycine 59·9 – – 52·3 – – 86·1 – –
Alanine 87·8 88·0 1·71 54·9a 64·8b 3·56 87·5 92·3 2·65
Overall mean2 84·9 86·8 2·14 71·4a 84·5b 1·96 89·8 93·3 2·12
1
Values are means of 6 replicate pens (two male and two female broilers per pen).
2
Mean of 15 amino acids (excluding ileal glycine digestibility values).
a,b
Within each ingredient, means in the same row with different superscripts are signiŽ cantly different (P < 0·05).

Table 5. Apparent amino acid digestibility (%)1 for meat-and-bone meal, Žsh meal and blood meal determined in the broiler chicken
Meat-and-bone meal Fish meal Blood meal
I.T. KADIM ET AL.

Ileal Excreta SEM Ileal Excreta SEM Ileal Excreta SEM

Threonine 69·2a 85·9b 1·09 84·6a 92·2b 2·25 55·4a 69·6b 6·12
Valine 79·7 74·6 2·25 86·8 87·4 2·98 59·2 58·8 5·17
Methionine 88·0 83·1 2·15 94·7 89·3 1·00 62·3 60·2 3·25
Isoleucine 71·0a 78·3b 1·89 87·5 87·8 1·52 – – –
Leucine 72·6a 81·2b 1·52 87·9 93·3 2·35 59·2 59·3 4·28
Phenylalanine 75·3 73·1 1·08 83·6 83·9 1·98 59·0 55·2 5·69
Tyrosine 69·5a 77·4b 1·98 86·7 85·5 3·25 48·4 45·1 4·25
Histidine 73·3 76·9 2·25 82·4 82·7 4·25 60·5 58·3 4·87
Lysine 77·4a 85·4b 2·05 93·4 94·7 1·52 65·8 69·0 4·02
Arginine 81·9 81·2 1·05 89·1 90·4 1·59 64·2 67·9 4·75
Aspartic acid 65·6a 82·7b 3·25 83·9 85·2 2·48 57·3a 74·2b 5·87
Serine 70·9a 78·2b 1·03 84·0 88·8 3·78 60·0 60·5 3·58
Glutamic acid 70·2a 80·2b 2·87 87·9 94·1 2·98 55·3a 65·6b 5·99
Proline 76·4 81·3 1·59 83·8 83·6 3·45 53·1 57·0 4·89
Glynine 86·0 – – 85·2 – – 53·7 – –
Alanine 71·7a 84·7b 1·59 88·5 87·2 2·89 57·4b 50·3a 4·35
Overall mean2 74·2 80·2 1·82 87·0 88·4 2·57 58·4 60·7 4·86
1
Values are means of 6 replicate pens (two male and two female broilers per pen).
2
Mean of 15 amino acids (excluding ileal glycine digestibility values). Mean of 14 amino acids for blood meal.
a,b
Within each ingredient, means in the same row with different superscripts are signiŽ cantly different (P < 0·05).
 Downloaded by [University of York] at 14:29 01 September 2013

Table 6. True amino acid digestibility (%)1 for sorghum, wheat and soyabean meal determined in the broiler chicken
Sorghum Wheat Soyabean meal
Ileal Excreta SEM Ileal Excreta SEM Ileal Excreta SEM

Threonine 91·6a 101·8b 5·24 91·3 89·0 3·12 94·7 98·9 3·25
Valine 99·1 103·2 3·67 82·9a 97·4b 3·25 92·7 96·4 3·78
Methionine 99·0a 112·2b 5·09 92·1a 101·4b 1·99 98·0 95·6 1·99
Isoleucine 98·1a 112·6b 4·97 88·3a 99·8b 2·25 96·2 96·3 2·00
Leucine 95·3 97·4 1·35 80·3a 99·9b 2·05 90·9 93·6 2·53
Phenylalanine 99·8 100·9 3·25 85·8a 94·9b 2·95 91·5 89·7 2·57
Tyrosine 97·5 104·3 1·99 77·6a 98·3b 3·25 91·7 96·1 2·58
Histidine 91·8 93·1 2·58 78·8a 97·6b 2·98 93·1 95·9 2·58
Lysine 98·3 104·0 1·99 90·6a 100·0b 2·25 95·7 96·7 1·00
Arginine 98·3 100·3 2·24 81·9a 99·1b 1·59 96·0 96·1 1·02
Aspartic acid 98·2 100·4 4·57 70·2a 109·1b 2·15 91·8 94·6 1·25
Serine 95·7 102·8 2·25 87·5 89·7 2·45 94·7 99·5 1·08
Glutamic acid 95·0 100·9 1·09 92·1 98·5 0·85 91·8 99·4 1·98
Proline 94·9 100·4 3·87 92·5 97·5 1·08 92·1a 100·6b 2·00
Glynine 86·9 – – 67·9 – – 89·7 – –
Alanine 94·5 101·4 3·01 59·4a 91·8b 3·59 90·8 96·8 2·58
Overall mean2 96·5b 102·4b 3·18 83·5a 97·6b 2·34 93·4 96·4 2·21
1
Values are means of 6 replicate pens (two male and two female broilers per pen).
2
Mean of 15 amino acids (excluding ileal glycine digestibility values).
a,b
Within each ingredient, means in the same row with different superscripts are signiŽ cantly different (P < 0·05).

Table 7. True amino acid digestibility (%)1 for meat-and-bone meal, Žsh meal and blood meal determined in the broiler chicken
Meat-and-bone meal Fish meal Blood meal

Ileal Excreta SEM Ileal Excreta SEM Ileal Excreta SEM

AMINO ACID DIGESTIBILITY

Threonine 85·5a 99·1b 1·99 93·9 99·9 2·87 62·7a 79·5b 5·25
Valine 83·6 84·9 3·25 92·1 94·3 3·05 62·1 64·2 4·82
Methionine 95·9 90·4 2·47 96·4 92·1 0·99 76·3 73·7 4·25
Isoleucine 79·8a 91·2b 3·00 92·4 94·2 1·69 – – –
Leucine 83·0 85·4 1·99 91·0 91·0 2·73 61·1 65·3 4·19
Phenylalanine 80·0 83·1 1·87 88·7 92·2 2·00 62·1 61·6 2·99
Tyrosine 73·6a 92·3b 1·59 91·0 91·8 2·49 54·6 57·4 7·59
Histidine 80·6 85·7 2·59 85·9 87·4 4·59 62·1 61·3 4·51
Lysine 88·3 90·9 1·23 95·5 97·3 2·59 67·7a 74·7b 4·28
Arginine 84·1 86·1 1·25 92·9 95·4 1·82 71·2 75·9 3·79
Aspartic acid 71·1a 91·7b 3·51 87·3 90·9 2·99 60·1a 78·9b 5·89
Serine 82·1 84·5 2·01 92·6 99·1 2·16 64·9 65·2 3·12
Glutamic acid 76·5a 88·9b 3·25 90·4a 100·4b 2·12 59·2 82·2 7·19
Proline 83·4 88·3 1·48 90·4 90·6 3·51 60·7a 66·4b 3·98
Glynine 91·9 – – 89·1 – – 59·1 – –
Alanine 84·5 91·1 1·98 91·3 93·0 0·98 59·7 59·2 3·49
Overall mean2 82·1 88·9 2·19 91·4 94·0 2·42 63·2 68·9 4·63
593

1
Values are means of 6 replicate pens (two male and two female broilers per pen).
2Mean of 15 amino acids (excluding ileal glycine digestibility values). Mean of 14 amino acids for blood meal.
a,b
Within each ingredient, means in the same row with different superscripts are signiŽ cantly different (P < 0·05).
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1
Table 8. Magnitude of the statistically signiŽcant (P < 0·05) excreta–ileal differences in apparent or true amino acid digestibility values of feed ingredients for broilers
Sorghum Wheat Soyabean meal Meat-and-bone meal Fish meal Blood meal

Apparent True Apparent True Apparent True Apparent True Apparent True Apparent True

Threonine +14·0 +11·1 +40·2 – +8·9 – +24·1 +15·9 +9·0 – +25·6 +26·8
Valine – – +16·3 +17·5 – – – – – – – –
Methionine – +13·3 – +10·1 – – – – – – – –
Isoleucine – +14·8 +24·2 +13·0 – – +10·3 +14·3 – – – –
Leucine – – +28·4 +24·4 – – +11·8 – – – – –
Phenylalanine – – +9·2 +10·6 – – – – – – – –
Tyrosine – – +24·3 +26·7 – – +11·4 +25·4 – – – –
Histidine – – +22·9 +23·9 – – – – – – – –
Lysine – – +11·8 +10·4 – – +10·3 – – – – +10·3
Arginine – – +29·0 +21·0 – – – – – – – –
I.T. KADIM ET AL.

Aspartic acid – – +55·6 +55·4 – – +26·1 +29·0 – – +29·5 +31·3

Serine +10·9 – +9·1 – – – +10·3 – – – – –
Glutamic acid – – – – – – +14·2 +16·2 – +11·1 +18·6 +38·9
Proline – – – – +9·1 +9·2 – – – – – +9·4
Alanine – – +18·0 +54·5 – – +18·1 – – – –12·4 –
1
(Excreta digestibility - ileal digestibility/ileal digestibility) ´ 100.

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 AMINO ACID DIGESTIBILITY 595

1 does not greatly affect the amino acid compo- mate the uptake of some amino acids from
2 sition of the non-dietary material voided in the some feeds. As can be seen from Table 8, the
3 ileal digesta or the excreta of birds fed on a degree of apparent overestimation was often
4 protein-free diet. However, signiŽ cantly higher considerable, ranging from 8·9% (apparent
5 outputs were found in the excreta for the digestibility of threonine in soyabean meal) to
6 dietary non-essential amino acids, aspartic acid 56% (apparent digestibility of aspartic acid in
7 and glutamic acid. Our results are in general wheat), and of such a magnitude to be of impor-
8 agreement with those of Bielorai and Iosif tance in practical dietary formulation. For one
9 (1987) who also reported similar amino acid amino acid (alanine in blood meal), a net
10111 patterns in the excreta and ileal digesta of appearance (presumably re ecting net synthe-
1 chicks fed on a protein-free diet. The endoge- sis) during passage through the hindgut was
2 nous amino acid outputs determined at the recorded. The difference remained even after
3 ileum of meat chickens in the present study correction was made for the contribution of
4 were comparable to those reported by Siriwan endogenous alanine.
5 et al. (1993). The determined digestibility values for the
6 The true ileal and excreta amino acid 6 ingredients assayed were, in general, within
7 digestibility data determined in our study do ranges already reported (Raharjo and Farrell,
8 indicate very high digestibilities of sorghum, 1984; Green et al., 1987a, b; NRC, 1994;
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9 wheat, soyabean meal and Ž sh meal, but dis- Ravindran et al., 1998). The relatively low appar-
20111 tinctly lower digestibilities of the proteins in ent digestibility for threonine in most feed
1 meat-and-bone meal and blood meal. However, ingredients as found in the present work is con-
2 for ingredients containing lower concentrations sistent with the results of other workers (Rahajo
3 of protein and amino acids (sorghum and and Farrell, 1984; Johns et al., 1986; Ravindran
4 wheat), correction for the endogenous compo- et al., 1999) and is likely be due to the relatively
5 nents led to true digestibility values exceeding high concentration of threonine in gut endoge-
6 100%. Clearly this is not possible and suggests nous protein. Another possibility is that the
7 that errors have arisen either in the method for peptide bonds involving threonine may be less
8 determining the dietary and digesta/excreta susceptible to breakdown by digestive enzymes.
9 amino acid  ows or in the determination of the Few studies which compare ileal digesta
30111 endogenous amino acid composition. The lim- with excreta-based determination of amino acid
1 itations of using a protein-free diet for measur- digestibility in chickens have been conducted.
2 ing endogenous amino acid losses have been That, in most instances, amino acids disap-
3 recently reviewed (Ravindran and Bryden, peared from the large intestine, is consistent
4 1999). The unexpectedly high true digestibility with other reports (Bielorai and Iosif, 1987; ten
5 values may have arisen because the endogenous Doeschate et al., 1993; Perez et al., 1993).
6 outputs measured in birds fed on the protein- Ravindran et al. (1999) reported apparent ileal
7 free diet differed from those fed on diets con- and excreta-based digestibility values of amino
8 taining adequate amounts of protein (Siriwan et acids for a wide range of feedstuffs. These
9 al., 1993). workers found that the amino acid digestibility
40111 The possible consequences of hindgut bac- in wheat was underestimated by excreta analy-
1 terial metabolic activity have prompted the sis. The relatively lower excreta digestibility
2 development of an ileal-based method for the values reported for wheat in their study is in dis-
3 determination of amino acid digestibility. In agreement with our results, where the excreta
4 the present study, signiŽ cant differences were digestibility values for wheat were found to be
5 found between ileal and excreta-based dig- markedly higher than the corresponding ileal
6 estibility coefŽ cients for the growing meat values. This discrepancy may be a result of
7 chickens, indicating a net catabolism of amino the differences in methodology employed to
8 acids in the large intestine. The degree of net measure amino acid digestibility and the rate of
9 amino acid disappearance was quite variable wheat inclusion in the assay diet. Ravindran et
50111 among ingredients and amino acids. When sig- al. (1999) used the direct method where wheat
1 niŽ cant (P < 0·05) differences were found, the was included at 918 g/kg and provided the sole
2 excreta digestibility coefŽ cient was usually source of protein in the assay diet. In the
3 higher than the ileal digestibility coefŽ cient. present study, the difference method was used,
4 These Ž ndings are suggestive of net disappear- with the assay diets containing 464 g/kg wheat.
5 ance of amino acids during passage from the Furthermore, the difference method assumes
6 ileaum to the end of the digestive tract. Because that digestibility coefŽ cients are additive, but it
7 absorption of amino acids is not thought to take is known that interactions between the dietary
8 place after the terminal ileum (Webb, 1990), ingredients may in uence digestibility estimates
9 the implication is that digestibility determina- (Kadim and Moughan, 1997b). In particular, it
60111 tions based on excreta collection will overesti- is likely that the arabinoxylans present in wheat
 596 I.T. KADIM ET AL.

may have lowered the digestibility of amino CO STIGAN , P. & ELLIS, K.J. (1987) Analysis of faecal chromium 1
acids in the basal diet by interfering with protein derived from controlled release marker devices. New 2
Zealand Journal of Technology, 3: 89–92.
digestion, increasing endogenous amino acid TEN DOESCHATE , R.A.H.M., S CH EELE , C.W., SCH REU RS ,
3
 ows or both (Angkanaporn et al., 1994). V.V.A.M. & VAN DER KILS, J.D. (1993) Digestibility studies 4
The net disappearance of amino acids from in broiler chickens: in uence of genotype, age, sex and 5
the large intestine was greater, in general, for method of determination. British Poultry Science, 34: 6
those with the lowest digestibility values at the 131–146. 7
FAN , M.Z. & SAUER , W.C. (1995) Determination of apparent
end of the small intestine. This is to be expected, ileal amino acid digestibility in barley and canola meal 8
since with lower digestibility at the ileum, signif- for pigs with the direct, difference and regression 9
icant quantities of undigested protein will reach methods. Journal of Animal Science, 73: 2364–2374. 10111
the large intestine providing substrate for micro- GREEN , S., SO LANGE , L., BERTRAND , L., MADELEINE , J., 1
bial degradation thus resulting in potentially DURON , J.C. & MAILLARD , R. (1987a) Digestibilities of 2
amino acids in maize, wheat and barley meal, determined
larger differences between ileal and excreta with intact and caecetomised cockerels. British Poultry 3
digestibility values. In general, threonine had Science, 28: 631–641. 4
the lowest digestibility at the ileum and, com- GREEN , S., SO LANGE , L., BERTRAND , L., MADELEINE , J., 5
pared with other amino acids, the highest DURON , J.C. & MAILLARD , R. (1987b) Digestibilities of 6
degradation during passage through the amino acids in soyabean, sun ower and groundnut 7
meals, determined with intact and caecetomised cock-
hindgut. erels. British Poultry Science, 28: 631–641. 8
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A potential criticism of the present work JO HNS, D.C., LO W, C.K. & JAM ES , K.A.C. (1986) Comparison 9
may be that the ileal measure of digestibility was of amino acid digestibility using the ileal digesta from 20111
made on the same bird, 3 d after the collection growing chickens and cannulated adult cockerels. British 1
of excreta. It is, however, considered unlikely Poultry Science, 27: 679–685. 2
KADIM , I.T. & MOU GHAN , P.J. (1997a) Development of an ileal
that the determined digestibility values were amino acid digestibility assay for the growing 3
in uenced by the duration of feeding of the chicken––effects of time after feeding and site of sam- 4
diet, since digestibility in the widely used rapid pling. British Poultry Science, 38: 89–95. 5
cockerel assay of Sibbald (1979) is measured fol- KADIM , I.T. & MO UG HAN , P.J. (1997b) Ileal amino acid 6
lowing a single meal. digestibility assay for the growing chicken––effects of the 7
imposition of a fasting period and the nature of the test
In conclusion, the present study has diet. British Poultry Science, 38: 285–290. 8
demonstrated signiŽ cant differences between LOW , A.G. (1990) protein evaluation in pigs and poultry, in: 9
ileal and excreta-based digestibility values of WISEM AN , J. & CO LE , D.J.A. (Eds) Feedstuff Evaluation, pp. 30111
certain amino acids in some feed ingredients 91–114 (London, Butterworths). 1
for growing meat chickens. Where differences MC NAB , J.M. (1994) Amino acid digestibility and availability 2
studies with poultry, in: D’MELLO , J.P.E. (Ed) Amino Acids
were found, amino acid digestibility was gener- in Farm Animal Nutrition, pp. 63–98 (Wallingford, CAB 3
ally higher when determined over the total International). 4
digestive tract. This observation suggests a sig- NRC (1994) Nutrient Requirements of Domestic Animals: Nutrient 5
niŽ cant net disappearance of amino acids in the Requirements of Poultry, 8th ed. (Washington, DC, National 6
large intestine, which is in accord with Ž ndings Research Council, National Academy Press). 7
PAPADO PO ULOS , M.C. (1985) Estimations of amino acid
in other simple-stomach species (Low, 1990). digestibility and availability in feedstuffs for poultry. 8
World’s Poultry Science Journal, 41: 64–71. 9
PEREZ , L., FERNAND EZ -FIG ARES, I., NIETO , R., AGU ILERA , J.F. 40111
ACKNOWLEDGEMENTS & PRIETO , C. (1993) Amino acid ileal digestibility of some 1
The Ž nancial support of Tegel Foods Ltd, grain legume seeds in growing chickens. Animal 2
Production, 56: 261–267.
Auckland, New Zealand and Australian Poultry RAH ARJO , Y. & FARRELL , D.J. (1984) A new biological method 3
Ltd, Sydney, Australia is gratefully acknowl- for determination of amino acid digestibility in poultry 4
edged. The technical assistance of Mr D.V. feedstuffs using a simple cannula, and the in uence of 5
Thomas is acknowledged with gratitude. dietary Ž bre of endogenous amino acid output. Animal 6
Feed Science and Technology, 12: 29–45. 7
RAVINDRAN , V. & BR YD EN , W.L. (1999) Protein evaluation in
8
REFERENCES poultry –– in vitro and in vivo measurements. Australian
Journal of Agricultural Research, 50: 889–908. 9
ANGKANAPORN , K., CH OCT, M., BRYDEN , W.L., ANNISO N , E.F. RAVINDRAN , V., HEW, L.I. & BR YDEN , W.L. (1998) Digestible 50111
& ANNISON , G. (1994) Effect of wheat pentosans on Amino Acids in Poultry Feedstuffs (Canberra, Rural 1
amino acid losses in chickens. Journal of the Science of Food Industries Research and Development Corporation and
and Agriculture, 66: 399–404.
2
Camden, Poultry Research Foundation, The University of
AOAC (1984) OfŽcial Methods of Analysis (Washington, DC, Sydney. 3
Association of OfŽ cial Analytical Chemists). RAVINDRAN , V., HEW, L.I., RAVIND RAN , G. & BRYDEN , W.L. 4
BAD AWY, A.M. (1964) Changes in the protein and non-protein (1999) A comparison of ileal digesta and excreta analy- 5
nitrogen in the digesta of the sheep, in: MU NRO , H.N. sis for the determination of amino acid digestibility in 6
(Ed) The Role of the Gastrointestinal Tract in Protein food ingredients for poultry. British Poultry Science, 40:
Metabolism, pp. 175–185 (Oxford, Blackwell).
7
266–274.
BIELORAI , R. & IOSIF, B. (1987) Amino acid absorption and SIBBALD , I.R. (1979) A bioassay for available amino acids and 8
endogenous amino acids in the lower ileum and excreta true metabolizable energy in feedingstuffs. Poultry Science, 9
of chicks. Journal of Nutrition, 117: 1459–1462. 58: 668–673. 60111
 AMINO ACID DIGESTIBILITY 597

1 SIBBALD , I.R. (1987) Estimation of bioavailable amino acids products: a review. Journal of Animal Science, 6: 3011–3022.
2 in feedingstuffs for poultry and pigs: a review with empha- WHITACRE , M.E. & TANNER , H. (1989) Methods of deter-
sis on balance experiments. Canadian Journal of Animal mining the bioavailability of amino acids for poultry, in:
3 Science, 67: 221–330. FRIEDMAN , M. (Ed) Absorption and Utilization of Amino
4 SIRIWAN , P., BRYDEN , W.L., MO LLAH , Y. & ANNISO N , E.F. Acids, Vol. III, pp. 129–141 (Boca Raton, FL, CRC Press).
5 (1993) Measurement of endogenous amino acid losses in YAP, K.H., KAD IM , I.T., KING , R.D. & MOU GHAN , P.J. (1997)
6 poultry. British Poultry Science, 34: 939–949. An ileal amino acid digestibility assay for the growing
7 SNEDECO R, G.W. & CO CH RAN , W.G. (1982) Statistical Methods meat chicken––effect of feeding method and digesta col-
(Ames, Iowa State University Press). lection process. Asian-Australasian Journal of Animal
8 WEBB , K.E. (1990) Intestinal absorption of protein hydrolysis Science, 10: 671–678.
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