Journal of the Science of Food and Agriculture J Sci Food Agric 80:547±554 (2000)

Low-protein amino acid-supplemented diets in

broiler chickens: effects on performance,
carcass characteristics, whole-body
composition and efficiencies of nutrient
utilisation
VA Aletor,* II Hamid, E Nieß and E Pfeffer
Institut für Tierernahrung, Universität der Bonn, Endenicher Allee 15, D-53115 Bonn, Germany

Abstract: Two concurrent trials were conducted to investigate the in¯uence of low-protein amino acid-
supplemented diets on the performance, carcass characteristics, whole-body composition and
ef®ciencies of nutrient utilisation by the male broiler chicken from age 3 to 6 weeks. The ®rst trial
comprised ®ve isoenergetic (13.0 MJ kgÿ1) diets containing 225 (control), 210, 190, 172 or 153 g kgÿ1
crude protein (CP) supplemented with essential amino acids (EAAs) to meet the minimum National
Research Council recommendations. In the second trial a composite mixture of non-essential amino
acids (NEAAs) was added to the lower-CP diets (ie 210±153 g kgÿ1) such that they became isoproteinous
(N  6.25) with the 225 g kgÿ1 control. Neither the lowering of dietary CP nor NEAA supplementation
had any signi®cant in¯uence on weight gain or the relative weights of the various carcass cuts.
However, chicks fed the lowest-CP diets consumed more feed (P  0.05) and had poorer (P  0.05) feed
conversion ef®ciency (FCE). NEAA supplementation enhanced FCE to the control levels. Whole-body
compositional analysis showed that lowering dietary CP increased (P  0.01) total body fat in a linear
fashion (P  0.001; r = ÿ0.72). Equalising dietary CP with the control (ie maintaining identical energy/
protein ratio) by NEAA supplementation did not correct for the fat deposition. Total body protein
(g kgÿ1) was identical with the control with or without NEAA supplementation. Dietary energy, protein
retention ef®ciency (PRE) and protein ef®ciency ratio (PER) were more ef®cient (P  0.01) in the
lower-protein diets, while NEAA supplementation signi®cantly (P  0.01) decreased the ef®ciency of N
utilisation. Reducing dietary CP from 225 to 153 g kgÿ1 decreased N excretion in a highly signi®cant
linear fashion (P  0.001; r = 0.73). The nutritional and environmental implications of the increased
body fat deposition on the one hand and the decreased N excretion on the other in the low-protein-fed
chickens are discussed and the need to harmonise these apparently con¯icting interests is emphasised.
# 2000 Society of Chemical Industry

Keywords: low-protein diets; amino acid supplementation; performance; whole-body composition; nutrient
utilisation; broiler chicken

INTRODUCTION to N pollution of the environment within the

The world is currently experiencing rapid rates of
urbanisation, industrialisation and agricultural devel-
opment. These activities are contributing signi®cantly
to the pollution of the environment in the form of
accidental chemical discharges, exhaust fumes, wastes,
sewage, by-products of manufacturing processes and
other human activities. Intensive animal production is
believed to be a major contributor to the pollution of
the environment. For example, in pigs, about 65% of
the ingested dietary nitrogen is excreted via urine and
faeces,1 and this general low ef®ciency of N utilisation
makes intensive animal production a main contributor
agricultural system.2
Within the European Economic Community
(EEC), for example, farm animal ef¯uents have been
identi®ed as a major source of nitrate pollution of
water systems. These environmental concerns about
ground water contamination with excess nitrate,
phosphorus as well as other elements resulted in the
issuance of Council Directive 91/670 EEC in 1991 in
an attempt to limit the amount of N in slurry that can
be spread on farm land.3 One of the current
approaches to reducing the risk of N pollution of the
environment is the reduction of N excretion in poultry

* Correspondence to: VA Aletor, Division of Nutritional Biochemistry, Division of Animal Production and Health, The Federal University of
Technology, PMB 704, Akure, Nigeria
Contract/grant sponsor: Alexander von Humboldt Foundation, Bonn
(Received 30 June 1999; accepted 1 October 1999)

# 2000 Society of Chemical Industry. J Sci Food Agric 0022±5142/2000/$17.50 547

VA Aletor et al
by dietary manipulations involving the feeding of low-
protein amino acid-supplemented diets. The overall
principle is to ensure a more ef®cient N utilisation by
supplementing low-protein diets with essential amino
acids (EAAs) in better agreement with standard
dietary recommendations. However, there are con-
¯icting reports on the effects of feeding low-protein
amino acid-supplemented diets to broiler chickens.
While some studies4±8 have reported impaired weight
gain and feed ef®ciency when broilers are fed low-
protein diets, others9,10 have reported identical per-
formance with conventional formulations. The most
consistent observation in these studies has been the
increased deposition of abdominal fat in low-protein-
fed chickens.
In all these investigations, there appear to be three
points with con¯icting interests: (i) the need to
maintain optimal/economic poultry productivity; (ii)
the need to maintain desirable (from the consumer
standpoint) carcass characteristics and composition;
and (iii) the compelling need to protect the environ-
ment via decreased N excretion. We believe that for
low-protein amino acid-supplemented diets to com-
mand wider commercial acceptability, current re-
search should attempt to harmonise these interests.
In this paper we present the results of two concurrent
trials in which broiler chickens were fed varying
protein levels supplemented with EAAs to meet
standard dietary recommendations. The lower-protein
diets were thereafter made isoproteinous with the
control by the supplementation of a composite mixture
of non-essential amino acids (NEAAs). In both cases,
performance, carcass characteristics, whole-body
composition and ef®ciencies of nutrient utilisation
(N excretion rates inclusive) were used as response
criteria.
MATERIALS AND METHODS
Experimental diets
The ®rst trial comprised ®ve isoenergetic
(13.0 MJ kgÿ1) broiler chick diets (D1±D5) containing
225, 210, 190, 172 or 153 g kgÿ1 crude protein
(N  6.25) formulated, on a least-cost basis, mainly
from maize/soya bean meal (SBM) mixtures (Table
1). To minimise any amino acid imbalances, the
maize/SBM ratio in all diets was kept constant at
1.22:1. The amino acid levels in all diets were
thereafter calculated and any de®cient EAAs were
supplemented in crystalline form to meet NRC
minimum recommendations (DL-methionine ‡ L-
cystine 7.2, L-lysine 10.0, L-threonine 7.4, L-trypto-
phan 1.8, L-arginine 12.0, L-valine 7.2, L-isoleucine
7.0, L-leucine 11.8, L-glycine ‡ L-serine 10.0 g kgÿ1).12
Necessary adjustments were made depending on the
purity of the supplements and such that all diets were
equal with respect to Ca, P and Na. The level of
dietary oil was kept relatively constant and the metab-
olisable energy (ME) and crude protein (CP) equiva-
548
lents of the supplemented amino acids were included
in the formulation as listed by the NRC.12
The second trial, which ran concurrently with the
®rst, was designed to ascertain the in¯uence of
NEAA supplementation on the low-protein diets. It
comprised four diets (D6±D9; Table 1) derived
from diets 2±5 by the respective addition of a
composite mixture of the preponderant NEAAs
(alanine, aspartic acid and glutamic acid). The
preponderance of these NEAAs in the feed was
ascertained from amino acid analysis. The NEAA
supplementation was such that diets 6±9 were both
isoenergetic and isoproteinous with diet 1, the control.
Again to minimise amino acid imbalances, the NEAAs
were supplemented in the same proportions (Ala/Asp/
Glu 1:2:3.4) as analysed to be present in the maize/
SBM mixtures. In all diets, corn starch, cellulose and,
to a lesser extent, oil levels were adjusted to equalise
dietary energy as dietary CP and amino acids were
altered. All diets were mechanically mixed and
pelleted before use.
Management of experimental chickens
Male Lohmann broiler (day-old) chicks were brooded
in a four-decked electrically heated battery brooder for
a 3 week pre-experimental period during which they
were fed a 23 g kgÿ1 protein broiler starter diet ad
libitum. The handling protocol of the chickens used
was consistent with that operational at the Poultry
Science Department and Alabama Agricultural Ex-
perimental Station, Auburn University, Alabama and
Canadian Council on Animal Care (CCAC). It
ensured proper care and treatment of the experimental
chickens. At the end of the 3 week pre-experimental
period, 13 chickens per group were randomly selected
from a batch of 150, weighed and randomly assigned
to each of the nine dietary treatments. The selection
and allocation procedure was such that the mean
group weights were identical (668.8  3.2 g). The
chicks were then wing-tagged and transferred to
three-tier individual grower metabolism cages of the
wire ¯oor type ®tted with automatic nipple drinkers.
They were fed the trial diets and provided with water
ad libitum until 6 weeks old, during which time the
weekly feed consumption and weight changes were
recorded. The trial was con®ned to the grower period
(3±6 weeks) as it represents the period of maximum
feed consumption and therefore cost as well as fat
deposition.
Organ and carcass measurements
At the end of the trial, all chickens were fasted
overnight to minimise interference from intestinal
content, then weighed. Those chickens (®ve per
treatment) having odd-numbered wing bands were
killed ®rst by stunning (35 V and 75 mA electric stun)
followed by bleeding and scalding.10 Similarly, those
chickens (®ve per treatment) with even-numbered
wing bands were killed by stunning/cervical disloca-
tion while avoiding loss of blood. They were then
J Sci Food Agric 80:547±554 (2000)
 Low-protein AA-supplemented diets in broiler chickens

Table 1. Compositions of experimental dietsa (g kgÿ1)

Ingredient D1 D2 D3 D4 D5 D6 D7 D8 D9
Maize (9.6%) 494.1 459.4 414.8 375.2 330.6 459.4 414.8 375.2 330.6
Soya bean meal (43.6%) 404.4 376.1 339.7 307.3 270.9 376.1 339.7 307.3 270.9
Corn starch Ð 50.3 115.0 171.1 231.9 33.5 75.3 111.0 150.9
Cellulose Ð 12.8 29.4 44.2 61.1 7.3 17.3 25.9 36.4
Soya oil 61.8 61.0 59.8 58.8 57.3 61.2 60.9 60.6 59.9
Limestone 5.1 5.2 5.3 5.4 5.5 5.2 5.3 5.4 5.5
DCP 17.8 17.9 18.1 18.3 18.5 17.9 18.1 18.3 18.5
Salt (NaCl) 3.6 3.6 3.7 3.7 3.7 3.6 3.7 3.7 3.7
Vitamin premixb 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0
Mineral premixb 6.0 6.0 6.0 6.0 6.0 6.0 6.0 6.0 6.0
Choline chloride 1.3 1.3 1.3 1.3 1.3 1.3 1.3 1.3 1.3
EAAs
Met ‡ cystine 1.9 2.4 2.9 3.3 4.0 2.4 2.9 3.3 4.0
.
Lysine HCl Ð Ð Ð 0.6 1.9 Ð Ð 0.6 1.9
Threonine Ð Ð Ð 0.7 1.4 Ð Ð 0.7 1.4
Tryptophan Ð Ð Ð 0.1 0.3 Ð Ð 0.1 0.3
Arginine Ð Ð Ð Ð 0.5 Ð Ð Ð 0.5
Valine Ð Ð Ð Ð 0.5 Ð Ð Ð 0.5
Isoleucine Ð Ð Ð Ð 0.6 Ð Ð Ð 0.6
NEAAs
Alanine Ð Ð Ð Ð Ð 3.4 8.0 12.0 16.1
Aspartic acid (1:2:3 4) . Ð Ð Ð Ð Ð 7.0 15.8 24.0 32.2
Glutamic acid Ð Ð Ð Ð Ð 11.7 26.9 40.6 54.8
Total 1000 1000 1000 1000 1000 1000 1000 1000 1000
Calculated analysis
Crude protein (g kgÿ1) 225.0 210.0 190.0 172.0 153.0 225.0 225.0 225.0 225.0
ME (MJ kgÿ1) 13.0 13.0 13.1 13.1 13.2 13.0 13.0 13.11 13.1
Energy/protein ratio (MJ kgÿ1) 58 62 68 76 86 58 58 58 58
Total Ca (g kgÿ1) 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6 8.6
Total P (g kgÿ1) 6.9 6.8 6.7 6.5 6.3 6.8 6.7 6.5 6.3
Available P (g kgÿ1) 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0
Na (g kgÿ1) 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5
a
Diets 1 (control)±5, ‡ EAAs; diets 6±9, ‡ EAAs ‡ NEAAs (derived from diets 2±5 respectively ‡ NEAAs).
b
Composition given elsewhere11 as follows. Vitamin mix supplied per kg diet: vit A, 5500 IU; vit D3, 11 000 IU; vit E, 11 mg; thiamin (B1), 3.5 mg; ribo¯avin (B2),
10.5 mg; Ca pantothenate, 11 mg; nicotinic acid, 70 mg; vit B6, 7 mg; folic acid, 1.75 mg; vit B12, 0.035 mg; choline chloride, 650 mg. Mineral mix supplied per kg
diet: Mn, 90 mg; Zn, 60 mg; Cu, 6 mg; Fe, 45 mg; Co, 0.15 mg; I, 0.45 mg; Se, 0.05 mg.

placed in individual cellophane bags and kept deep- Chemical analyses

frozen at ÿ20 °C prior to whole-body analysis. Again,
whenever it was necessary, adjustments were made
such that the mean group weights for all samples were
identical. The carcasses for dissection were scalded at
80 °C in a water tank for about 30 s before mechanical
plucking of the feathers. After warm washing, draining
and cooling, the carcasses were eviscerated. The
dressed and eviscerated weights were recorded before
carefully dissecting out the breast, drumstick, thigh
and wing. The corresponding organsÐliver, spleen,
heart, pancreas, gizzard, lungsÐand abdominal fat
were carefully dissected out and blotted free of blood
before weighing. In the present trial the abdominal fat
pad is de®ned as the tissue surrounding the gizzard
and intestines, extending within the ischium, and
surrounding the cloaca, bursa of fabricus and adjacent
to the abdominal muscles.6 All dissections were done
on the same day and by the same person to ensure
uniformity of cuts.
Amino acid composition of feed
Prior to the dietary formulations the CP and amino
acid compositions of the maize and SBM (solvent-
extracted) were analysed. Similarly, after formulation
and pelleting, all diets were milled to pass through a
1 mm sieve and the amino acids determined after HCl
hydrolysis by the Degussa Corporation, Hanau,
Germany. The calculated and analysed values, which
were largely similar, are shown in Table 2.
Whole-body compositional analysis
Prior to analysis the carcasses were thawed, transferred
into individual tightly sealed copper cans wrapped in
cellophane bags and autoclaved at 110 °C at 1 atm for
5 h. Thereafter the cans were cooled and the carcasses
(including feathers) transferred (avoiding loss of
materials) into an Edward Muller & Sohne homo-
geniser (Model MTZ, 10/70, Saarbrucken). After
thorough homogenisation, about 10 g each was taken

J Sci Food Agric 80:547±554 (2000) 549

VA Aletor et al

D1 D2 D3 D4 D5 D6 D7 D8 D9
ME (MJ kgÿ1) 13.0 13.0 13.1 13.1 13.2 13.0 13.0 13.1 13.1
CP (g kgÿ1) 225 210 190 172 153 225 225 225 225
(227) (210) (194) (175) (16) (225) (225) (224) (224)
Methionine ‡ cystine 8.8 8.8 8.7 8.5 8.6 8.8 8.7 8.5 8.6
(8.7) (8.5) (8.4) (8.2) (8.2) (8.6) (8.5) (8.4) (8.3)
Lysine 12.6 11.7 10.6 10.1 10.0 11.7 10.7 10.1 10.0
(12.3) (11.1) (10.1) (9.5) (9.5) (11.3) (10.2) (9.8) (9.7)
Threonine 9.0 8.3 7.5 7.5 7.4 8.3 7.6 7.6 7.4
(8.7) (7.9) (7.2) (7.2) (7.1) (8.1) (7.3) (7.3) (7.0)
Tryptophan 2.4 2.2 2.0 1.9 1.9 2.2 2.0 1.9 1.9
(2.8) ND ND ND (2.1) ND ND ND (2.1)
Arginine 16.1 15.0 13.6 12.3 11.3 15.0 13.7 12.3 11.3
(15.6) (14.2) (12.9) (11.6) (10.8) (14.4) (13.0) (11.9) (11.0)
Valine 11.6 10.8 9.7 8.8 8.2 10.8 9.8 8.8 8.2
(10.5) (9.6) (8.8) (8.0) (7.6) (9.7) (9.0) (8.1) (7.6)
Isoleucine 10.1 9.4 8.5 7.6 7.3 9.4 8.5 7.6 7.3
(9.9) (9.0) (8.3) (7.6) (7.2) (9.2) (8.3) (7.5) (7.3)
Leucine 20.2 18.8 17.0 15.4 13.6 18.8 17.1 15.4 13.6
(20.2) (18.4) (17.1) (15.6) (13.8) (18.9) (17.1) (15.4) (13.6)
Histidine 6.8 6.3 5.7 5.2 4.6 6.3 5.7 5.2 4.6
(6.6) (6.1) (5.4) (5.0) (4.4) (6.1) (5.5) (5.0) (4.4)
Glycine ‡ serine 21.7 20.2 18.2 16.5 14.6 20.2 18.4 16.5 14.6
Table 2. Calculated and analysed
(20.8) (19.0) (17.9) (15.7) (14.0) (19.2) (17.4) (15.8) (14.0)
amino acid contents (g kgÿ1) of
experimental diets Values in parentheses are analysed. ND, not determined.

for dry matter and total ash determinations. Similarly, estimated:

about 500 g of each homogenate was dried to constant
energy retention efficiency…%† ˆ
weight for about 3 days using a Leybold Heraeus
freeze-dryer. Following freeze-drying, the samples MJ energy retained
 100
were then milled before analysis. Six chickens MJ energy consumed
(selected at the start of the experiment) with identical
protein retention efficiency…%† ˆ
mean weight were similarly processed and analysed to
permit protein and fat retention estimates. The dry g protein retained
 100
matter and total ash in the carcasses were determined g protein consumed
according to AOAC methods,13 while the protein was g weight gain
determined after Kjeldahl digestion using a Gerhardt protein efficiency ratio ˆ
g protein consumed
automatic N analyser (Type VAP 6, Bonn). The total
body fat was determined by the technique of LUFA.14 lysine retention efficiency…%† ˆ
This entailed the pre-extraction of a 3 g sample g lysine retained
wrapped in a glass thimble for 10 min with anhydrous g lysine consumed  100
diethyl ether using a Soxhlet extraction unit. The
sample was then dried for 30 min and the fat content N excretion (apparent) ˆ
recorded. The pre-extracted sample plus 3 g Celite was g N consumed ÿ g N retained
then hydrolysed with 100 cm3 3 M HCl for 30 min
followed by dilution with 100 cm3 cold distilled water.
Each tube was then washed with hot distilled water Data analysis
and dried overnight at 70 °C. Following hydrolysis, the All data were subjected to analysis of variance
samples were again transferred to the fat extraction (ANOVA) and regression analyses using the SAS
unit and extracted with diethyl ether in the `boiling' statistical package.15 Where treatment means were
position for 60 min. The samples were then removed signi®cantly different, they were compared using
and dried at 95±100 °C for 30 min, cooled in a Duncan's multiple-range test.16
desiccator and the fat content calculated. The total
fat was the summation of the pre-extraction and ®nal
extraction. RESULTS
Performance
The weight gain (WG), feed consumption (FC) and
Estimates of efficiencies of nutrient utilisation feed conversion ef®ciency (FCE) are presented in
After chemical analysis, the following parameters were Table 3. Reducing dietary CP from 225 to 153 g kgÿ1

550 J Sci Food Agric 80:547±554 (2000)

 Low-protein AA-supplemented diets in broiler chickens

Table 3. Performances of broiler chickens fed low-protein amino acid-supplemented diets

D1 D2 D3 D4 D5 D6 D7 D8 D9
ÿ1
CP (g kg ) 225 210 190 172 153 225 225 225 225
ME (MJ kgÿ1) 13.0 13.0 13.1 13.1 13.2 13.0 13.0 13.1 13.1
Weight gain (g per chick) 1522 1503 1597 1472 1539 1536 1495 1541 1525
 204 173 80 209 114 183 194 131 161
Feed consumption (g per chick) 2721bc 2755abc 2932ab 2771abc 2960a 2688c 2695c 2851abc 2754abc
 285 225 149 278 203 287 249 170 330
Feed conversion (feed/gain) 1.80bc 1.84abc 1.84abc 1.89ab 1.92a 1.75c 1.81bc 1.82abc 1.81bc
 0.19 0.15 0.07 0.17 0.07 0.07 0.1 0.07 0.08
Values are mean  SD for 13 replications per diet. Means without common superscripts in the same row differ signi®cantly (P  0.05).

had no signi®cant in¯uence on weight gain. NEAA
supplementation to the diets to equal the CP level of
the control also had no effect on WG. However, the
chickens consumed more feed (P  0.05) and had
poorer FCE with decreasing CP. Chickens fed NEAA-
supplemented diets had improved FCE comparable
with the control.
Relative organ weights and carcass characteristics
The relative organ weights (g kgÿ1 body weight) for all
dietary treatments had the following rangesÐliver
17.9±20.7, spleen 0.9±1.3, heart 5.1±6.9, pancreas
1.6±1.9, gizzard 9.1±12.8, lungs 5.5±7.4, abdominal
fat 16.8±22.9Ðwhile the ranges for the various carcass
cuts were: dressed weight % 86.1±88.2, eviscerated
weight % 66.9±68.7; breast 190.5±202.9, drumstick
42.1±47.9, thigh 52.4±55.6, wing 35.2±37.3 (g kgÿ1
body weight). Decreasing dietary CP generally tended
to increase the relative weights of the liver and
abdominal fat, although such increases were not
signi®cant (P  0.05). The supplementation of NEAAs
had no signi®cant effect on the organ weights. Like the
organ weights, neither the lowering of dietary CP nor
NEAA supplementation signi®cantly in¯uenced the
dressed weight (%), eviscerated weight (%) or the `fast
food' cutsÐbreast, drumstick, thigh and wing.
Whole-body composition
Whole-body compositions (g kgÿ1 fresh weight) of the
treatment groups are presented in Table 4. The dry
matter (DM) and total body fat increased signi®cantly
(P  0.01) with decreasing dietary CP. Regression
analysis (see Table 7) of DM or total body fat
(dependent variable) against CP consumed (indepen-
dent variable) showed highly signi®cant negative
relationships (P  0.01; r = ÿ0.62 and P  0.001;
r = ÿ0.72 respectively). Total body ash was also
signi®cantly in¯uenced (P  0.01) in a manner not
consistently related with dietary treatments. Total
body protein content was identical across all dietary
treatments. Total body fat content was consistently
increased (P  0.01) irrespective of NEAA supplemen-
tation.
Protein and fat retention
Protein retention (Table 5) tended to increase (though
non-signi®cantly) across dietary treatments when
compared with the control. Conversely, decreasing
dietary CP signi®cantly (P  0.001) increased fat
retention (deposition) from about 146 g fat per chick
in the 225 g kgÿ1 CP diet to 289 g fat per chick in the
153 g kgÿ1 CP diet. This represents a twofold increase
in fat deposition. Regressing fat retention (Y) against
CP consumed (X) indicated a very highly signi®cant
negative relationship with the following prediction
equation: Y = 427.9 ÿ 0.42X (P  0.001; r = ÿ 0.68).
Surprisingly, equalising the dietary CP with the
control by NEAA supplementation (ie maintaining
identical energy/protein ratio) did not correct for
increased fat retention (deposition). For example,

Table 4. Whole-body compositions of broiler chickens fed varying protein levels supplemented with amino acids

D1 D2 D3 D4 D5 D6 D7 D8 D9
ÿ1
CP (g kg ) 225 210 190 172 153 225 225 225 225
ME (MJ kgÿ1) 13.0 13.0 13.1 13.1 13.2 13.0 13.0 13.1 13.1
Dry matter (g kgÿ1) 351.8bc 350.2bc 344.2bc 361.2b 381.6a 339.4c 339.8c 356.4bc 356.4bc
 11.7 8.2 8.6 5.3 6.6 16.2 10.4 5.5 23.1
Total body fat (g kgÿ1) 107.6d 108.5d 124.6c 132.2bc 162.7a 108.1d 123.8c 140.4b 154.4a
 6.2 6.7 6.3 5.4 8.1 6.8 2.7 5.5 16.7
Total body protein (g kgÿ1) 198.0 189.7 182.1 189.1 187.3 197.5 186.2 183.8 182.7
 5.1 4.2 4.2 2.9 8.1 6.8 9.1 4.6 6.1
Total body ash (g kgÿ1) 25.8a 25.2ab 23.7bc 24.6abc 25.2ab 25.0ab 24.8abc 22.9dc 21.4d
 1.7 0.7 1.4 1.5 1.2 1.2 1.3 1.5 2.5
Values are mean  SD for ®ve replications per diet. Means without common superscripts in the same row differ signi®cantly (P  0.01).

J Sci Food Agric 80:547±554 (2000) 551

VA Aletor et al

Table 5. Protein and total fat retention in broiler chickens fed varying protein levels supplemented with amino acids (n = 5)

D1 D2 D3 D4 D5 D6 D7 D8 D9
Protein retention (g per chick) 268.9 306.3 288.1 281.5 285.8 305.6 274.3 287.0 279.0
 45.6 10.6 7.0 26.9 12.9 35.0 25.0 7.3 40.4
Fat retention (g per chick) 146.0d 180.1dc 215.3bc 217.1bc 288.9a 168.4d 186.8dc 249.8ab 277.4a
 24.0 19.4 16.0 17.3 32.1 21.6 32.0 23.0 47.0
Means without common superscripts in the same horizontal row differ signi®cantly (P  0.01).

Table 6. Efficiencies of energy and protein (N  6.25) utilisation in broiler chickens fed varying protein levels supplemented with amino acids (n = 5)

D1 D2 D3 D4 D5 D6 D7 D8 D9
ÿ1
CP (g kg ) 225 210 190 172 153 225 225 225 225
ME (MJ kgÿ1) 13.0 13.0 13.1 13.1 13.2 13.0 13.0 13.1 13.1
Energy retention ef®ciency (%) 38.8d 37.2d 38.6cd 40.8bc 46.0a 38.6cd 39.4cd 42.8b 47.0a
 2.9 1.5 1.6 0.7 3.1 2.2 2.9 1.8 3.1
Protein retention ef®ciency (%) 48.2bc 50.8b 50.6b 58.2a 61.5a 50.6b 45.2c 44.2c 44.6c
 4.9 3.3 2.6 1.8 4.5 1.5 4.6 0.8 1.5
Protein ef®ciency ratio (PER) 2.5c 2.6c 2.9b 3.0b 3.4a 2.5c 2.5c 2.4c 2.5c
 0.2 0.2 0.2 0.3 0.1 0.1 0.2 0.1 0.1
Lysine retention ef®ciency (%) 41.4 46.0 45.3 50.3 48.3 47.0 49.6 47.4 48.2
 4.7 2.8 1.9 1.3 3.4 1.0 3.7 0.7 1.4
Nitrogen excretion (g per chick) 48.1bc 47.1bc 44.6c 37.7d 28.2d 47.1bc 46.9bc 56.9a 54.7ab
 7.2 5.8 3.3 2.3 4.8 4.8 8.1 2.2 8.5
Means without common superscripts in the same horizontal row differ signi®cantly (P  0.01).

chicks fed diet 5 or 9 retained similar amounts of fat
(289 vs 277 g per chick) as compared with the control
(146 g per chick).
Efficiencies of energy and protein (N  6.25)
utilisation
Table 6 presents data on energy retention ef®ciency
(ERE), protein retention ef®ciency (PRE), protein
ef®ciency ratio (PER), lysine ef®ciency ratio (LRE)
and apparent N excretion. ERE, PRE and PER were
generally signi®cantly (P  0.01) improved with de-
creasing dietary CP. Regression of ERE, PRE or PER
against CP consumption showed signi®cant (P  0.01)
negative correlations with respective r values of ÿ 0.64,
ÿ 0.70 and ÿ 0.88. NEAA supplementation generally
lowered PRE and PER values in comparison with the
unsupplemented counterparts. Lysine retention was
generally increased (though non-signi®cantly) when
compared with the control.
In contrast with fat retention, N excretion was
decreased by 41% as dietary CP was decreased.
Regression analysis (Table 7) showed a very highly
signi®cant (P  0.001) positive correlation (r = 0.73)
between N excretion (Y) and CP consumption (X) as
described by the following equation: Y =
ÿ 2.4 ‡ 0.08X. NEAA supplementation increased N
excretion in amounts comparable with (and in some
cases exceeding) the control values.
DISCUSSION
The present study demonstrates that the growth of the
grower broiler chicken is unaffected by decreasing
dietary CP from 225 to 153 g kgÿ1 when such diets are
supplemented with EAAs that meet the minimum

Table 7. Simple regression and correlation between whole-body composition, efficiencies of nutrient utilisation and crude protein consumption (n = 5)

Parameters Prediction equation Correlation coef®cient Signi®cance level

Dry matter (Y) vs CP consumed (X) Y = 416.9 ÿ 0.11X r = ÿ0.62 **
CP whole-body (Y) vs CP consumed (X) Y = 180.7 ‡ 0.02X r = ‡0.17 NS
Total fat (Y) vs CP consumed (X) Y = 221.9 ÿ 0.18X r = ÿ0.72 ***
Ash content (Y) vs CP consumed (X) Y = 23.9 ‡ 0.002X r = ‡0.12 NS
CP retained (Y) vs CP consumed (X) Y = 296.1 ÿ 0.019X r = ÿ0.06 NS
Fat retained (Y) vs CP consumed (X) Y = 427.9 ÿ 0.42X r = ÿ0.68 ***
Energy retention ef®ciency (Y) vs CP consumed (X) Y = 55.3 ÿ 0.029X r = ÿ0.64 **
Protein retention ef®ciency (Y) vs CP consumed (X) Y = 79.9 ÿ 0.05X r = ÿ0.70 ***
Lysine retention ef®ciency (Y) vs CP consumed (X) Y = 59.7 ÿ 0.026X r = ÿ0.54 **
N excretion (Y) vs CP consumed (X) Y = ÿ2.49 ‡ 0.08X r = ‡0.73 ***
**, P  0.01.
***, P  0.001; NS, not signi®cant.

552 J Sci Food Agric 80:547±554 (2000)


NRC speci®cations.12 It also shows that equalising the
CP levels by NEAA supplementation had no effect on
the rate of growth. However, the increased feed
consumption in the low-protein diets led to a
corresponding decrease in feed conversion ef®ciency.
The only advantage of NEAA supplementation on
performance appears to be related to its tendency to
reduce FC, with the resultant improvement in FCE
comparable with the control (Table 3). These ®ndings
generally agree with earlier studies,5±8 but contrast
with those of Paars and Summers9 and Moran et al,10
who reported that such low-protein diets failed to
support identical live weight gain as the control. The
discrepancies in responses often observed in the
literature in chicks fed low-protein amino acid-
supplemented diets appear to be related to, among
other factors, the degree of reduction of CP levels, the
inclusion or otherwise of the CP and ME contribu-
tions of the amino acid supplements, the class and age
of the chickens used and whether or not the intact
protein sources are kept at constant ratios to minimise
amino acid imbalances. In this study we have used
male broilers, included the CP and ME contributions
of the amino acid supplements in the formulations and
kept the maize/SBM ratios and dietary ME levels
constant, thus permitting a more accurate comparison
between treatment groups.
The most consistent observation in chicks fed low-
protein amino acid-supplemented diets has been
increased feed consumption with a concomitant
increase in carcass fat deposition.6,7,17,18 The present
study indicates that decreasing dietary CP generally
tended to increase the relative weights of the liver and
abdominal fat, although such increases were not
signi®cantly different. The liver is the primary site of
lipogenesis in chickens,17 suggesting that the tendency
for increased liver weight in the low-protein diets may
be related to increased lipogenetic activity. Female
broilers have been shown to accumulate more abdom-
inal fat than male broilers,19,20 suggesting that the lack
of signi®cance in the present study in comparison with
previous studies feeding similar protein levels6 may be
related to sex. NEAA supplementation appeared to
marginally reduce abdominal fat deposition in com-
parison with the low-protein-fed chicks. Neither the
lowering of CP nor NEAA supplementation had any
signi®cant in¯uence on carcass `fast food' cuts,
suggesting that identical carcass traits are attainable
by feeding 225 or 153 g kgÿ1 CP.
Whole-body compositional analysis suggests that
decreasing dietary CP to the extent used in this study
has no signi®cant in¯uence on total body protein
content, while protein retention (deposition) within
the period increased non-signi®cantly. Conversely,
total body fat content was increased by 51%, while fat
retention during the period was doubled. The fact that
abdominal fat was not signi®cantly increased in spite
of a twofold increase in total fat retention suggests that
abdominal fat may not be a good index of carcass
fatness in male broilers. It is generally believed
J Sci Food Agric 80:547±554 (2000)
Low-protein AA-supplemented diets in broiler chickens
that diets containing large energy/protein ratios
(> 72 MJ kgÿ1 CP (N  6.25)) promote high rates of
in vitro lipogenesis17 as well as de novo carcass lipid
synthesis by the chicken liver,21,22 while diets with
small energy/protein ratios (<56 MJ kgÿ1 CP) promote
lean carcasses. The offered interpretation is that large
energy/protein ratios force over-consumption of diet-
ary energy, as broilers eat to meet dietary protein/
amino acid requirements. Conversely, diets containing
small calorie/protein ratios are believed to decrease
carcass fat, because broilers consume less energy in
meeting protein requirements. According to Buttery
and Boorman23 and Bartov,24 the primary mechanism
involved in the reduction of carcass fatness by feeding
high-CP diets is the associated increased energy
expenditure and increased heat increment in degrad-
ing excess amino acids to uric acid. The present study
suggests that the above hypotheses do not fully explain
the increased carcass deposition in low-protein amino
acid-supplemented diets. If energy/protein ratio per se
were to be responsible, then diets 1 (control) and diets
6±9 (Table 1) containing identical energy/protein
ratios (58 MJ kgÿ1 CP) should have identical fat
content or identical fat retention (Tables 4 and 5
respectively), which was not the case. A more likely
interpretation appears to be the in¯uence of energy/
intact protein ratio rather than energy/protein
(N  6.25) ratio per se. This is evident in this trial, as
only those chicks consuming diets with identical
energy/intact protein ratio (rather than energy/protein
ratio per se) generally had similar fat contents (Table
4). Similarly, if increased energy expenditure asso-
ciated with increased N excretion were to fully account
for the decreased carcass fatness as suggested by
Buttery and Boorman23 and Bartov,24 then chicks fed
diet 1 (control) excreting identical amount of N (Table
6) should have identical carcass fatness as diets 6±9.
Again, this was not the case, clearly suggesting that
crystalline (synthetic) amino acids may in¯uence
lipogenesis differently from protein-bound (natural)
amino acids. It is unclear whether the difference is
related to the differences in the absorption rates
between peptides and monomeric amino acids.
The low-protein diets generally utilised dietary
energy and protein more ef®ciently than the control,
while NEAA supplementation led to poorer PRE and
PER values in agreement with previous reports.6,7,25
This clearly suggests that for the most ef®cient
utilisation of protein, diets should be formulated to
furnish intact protein and essential amino acids in
amounts no greater than required for optimum
performance, since N consumed in excess of require-
ment is degraded and excreted.
CONCLUSIONS
While the present study has generated data that
address impacts on economic, environmental and
acceptability concerns, it is dif®cult to recommend a
single diet which integrates the optimisation of these
553
VA Aletor et al
important aspects of broiler production. However, it is
evident that feeding low-protein amino acid-supple-
mented diets (eg 153 g kgÿ1 CP) confers potential
economic advantage, while the accompanying in-
creased ef®ciency of N utilisation lowers the amount
of excretable N. The reduction of N excretion by 41%
in the 153 g kgÿ1 CP diet suggests that dietary
formulations based on similar principles could con-
tribute towards alleviating current environmental
concerns regarding acid rains and ground water
pollution with excess nitrogen. However, there still
remains the problem of increased carcass fat deposi-
tion in low-protein-fed chickens. The need for further
studies to address this problem of carcass fatness is
compelling because of the current acute consumer
awareness of the association between dietary animal
fat consumption and human cholesterol-related ail-
ments.
ACKNOWLEDGEMENTS
The ®rst author wishes to express his gratitude to the
Alexander von Humboldt Foundation, Bonn for the
Fellowship award to carry out this study. He would
also like to thank all members of staff and postgraduate
students of the Institut fuÈr Tierernahrung for their co-
operation. We would also like to thank Drs Pack and
Koch, Degussa AG, Hanau for assisting in the amino
acid analysis. Many thanks are due to Dr D Tuschy for
his assistance in the statistical analysis.
REFERENCES
1 Scat JD, de Jong J and Kempen JP, Dietary protein in relation to
requirement and pollution in pigs during the body weight range
of 20 to 40 kg. Proc 1st Int Symp on N-¯ow in Pig Production and
Environmental Consequences, pp 259±263 (1993).
2 Kirchgessner M and Roth FA, Minderung der Stickstoff-
Ausscheidung beim Schwein. Arch Amin Nutr 43:283±301
(1993).
3 European Economic Community (EEC), Concerning the pro-
tection of water against pollution caused by nitrate from
agricultural sources. Council Directive 91/676/EEC, December
(1991).
4 Uzu G, Limits of reduction of the protein level in broiler feed.
AEC Information No 242, Commentary 03600, Rhone-Poulence
Corp (1983).
5 Summers JD and Leeson S, Broiler carcass composition as
affected by amino acid supplementation. Can J Anim Sci
65:717±723 (1985).
6 Fancher BI and Jensen LS, In¯uence on performance of three to
six-week-old broilers of varying dietary protein contents with
supplementation of essential amino acids requirements. Poultry
Sci 68:113±123 (1989).
554
7 Fancher BI and Jensen LS, Male broiler performance during the
starting and growing periods as affected by dietary protein,
essential amino acids, and potassium levels. Poultry Sci
68:1385±1395 (1989).
8 Holsheimer JP and Janssen WMMA, Limiting amino acids in
low-protein maize±soyabean meal diets fed to broilers from 3±7
weeks of age. Br Poultry Sci 32:151±158 (1991).
9 Paars JF and Summers JD, The effect of minimizing amino acids
excesses in broiler diets. Poultry Sci 70:1540±1549 (1991).
10 Moran ET, Bushong RD and Biligi SF, Reducing dietary crude
protein while satisfying dietary amino acids by least-cost
formulation: live weight performance, litter composition, and
yield of fast food carcass cuts at six weeks. Poultry Sci 71:1687±
1694 (1992).
11 Vogt H, Einsatz von Phytase im Broiler Mastfutter mit
unterschiedlichen Phosphorgehalt: 2. Versuch. Arch Ge¯ugelk-
unde 56:222±226 (1992).
12 National Research Council (NRC), Nutrient Requirements of
Domestic Animals. 1. Nutrient Requirements of Poultry, National
Academy Press, Washington, DC (1984).
13 AOAC, Of®cial Methods of Analysis, 13th edn, Association of
Of®cial Analytical Chemists, Washington, DC (1985).
14 Verband Deutscher Landwirtscha¯icher Untersuchungs- und
Forschungsanstalten (LUFA), Methodenbuch, Band III. Die
Chemische Untersuchung von Futtermitteln, Verlag Neumann,
Neudamm, pp 1±2 (1993).
15 Schuener R, Strolein G and Gogolok J (Eds), Datenverarbeitung
und Statistiche Auswertung mit SAS. Band II: Komplexe
Statistische Analyseverfahren, Gustav Fischer Verlag, Stuttgart
(1990).
16 Steel RGD and Torrie JH, Principles and Procedures of Statistics,
McGraw-Hill, New York (1960).
17 Rosebrough RW and Steele NC, Energy and protein relation-
ships in broilers. 1. Effect of protein levels and feeding
regimens on growth, body composition and in vitro lipogenesis
of broiler chicks. Poultry Sci 64:199±126 (1985).
18 Rosebrough RW and McMurtry JP, Protein energy relationship
in broiler chickens. 11. Effect of protein quantity and quality
on metabolism. Br J Nutr 70:667±678 (1993).
19 Rinehart KE, Green DE and Williamson JL, The in¯uence of
selected nutrition and management factors on broiler carcass
composition. Poultry Sci 54:1809±1810 (1975).
20 Twinning PV, Thomas OP and Bassard EH, Effect of diet and
type of bird on carcass composition of broilers at 28, 49, 59
days of age. Poultry Sci 57:492±497 (1978).
21 Grif®ths L, Leeson S and Summers JD, Fat deposition in
broilers: effects of dietary energy to protein balance, and early
life calorie restriction on production performance and abdom-
inal fat pad size. Poultry Sci 56:638±646 (1977).
22 Donaldson WE, Lipogenesis and body fat in chickens. Protein:-
calorie ratio and dietary fat. Poultry Sci 64:1199±1204 (1985).
23 Buttery PJ and Boorman KN, The energetic ef®ciency of amino
acids metabolism. In Protein Metabolism and Nutrition, Ed by
Cole DJA, Butterworths, London, pp 56 (1976).
24 Bartov I, Nutritional factors affecting quantity and quality of fat
in chickens. Fed Proc 38:2627±2639 (1979).
25 Wouldroup PW, Mitchell RJ, Payne JR and Hazen KR,
Performance of chicks fed diets formulated to minimize excess
levels of essential amino acids. Poultry Sci 55:243±253 (1976).
J Sci Food Agric 80:547±554 (2000)