Effects of the Naked Neck (Na) Gene on the Sulfur-Containing
Amino Acid Requirements of Broilers1
G. M. PESTI,2 B. LECLERCQ, A.-M. CHAGNEAU, and T. COCHARD
Station de Recherches Avicoles, Institut Nacional de la Recherche Agronomique, 37380 Nouzilly, France
ABSTRACT Full-sibling normal (na/na) and naked
neck (Na/na) chickens were fed from 28 to 42 d of age on
one of five diets with different SAA contents (from 5.4 to
7.0 g/kg). The experimental diets were made by adding
DL-methionine to a well-balanced corn, soybean, and
cornstarch diet containing 5.4 g SAA/kg and 161 g
crude protein/kg. Dietary SAA influenced the growth
rate of both genotypes similarly. There were no
significant differences (P > 0.125) in body weight gain
due to genetics or a diet by genetics interaction. Body
weight gains were maximized at 6.24 ± 0.45 (R2 = 0.171)
and 5.96 ± 0.52 g / k g SAA (R2 = 0.107) for the na/na and
Na/na stocks, respectively. There was a significant SAA
by genotype interaction for feed efficiency: the na/na
birds were more efficient at low SAA levels, but the Na/
(Key words: sulfur-containing amino acids, naked neck, broiler)
INTRODUCTION
Leclercq et al. (1993) demonstrated that lean line (LL)
chickens from the Institut Nacional de la Recherche
Agronomique required higher dietary concentrations of
SAA for maximum growth than their fat line (FL)
chickens. The LL birds had lower feed consumption and
greater feather synthesis, and used the SAA more
efficiently than the FL chickens. Leclercq et al. (1993)
concluded that LL chickens require diets with higher
concentrations of SAA because they consume less feed
and allocate a greater proportion of SAA to feather
synthesis.
Leclercq and Guy (1991) demonstrated that LL and
FL chickens have different protein requirements as-
sociated with differences in both carcass composition
and feathering of the two lines. Ajang et al. (1993)
observed feather protein gain and protein requirements
with birds selected for fast and slow feathering. The data
of Ajang et al. (1993) suggested that the degree of
feathering may significantly influence the protein re-
Received for publication June 15, 1995.
Accepted for publication November 8, 1995.
Supported by INRA and state and Hatch funds allocated to the
Georgia Agricultural Experiment Stations of the University of Georgia.
2
To whom correspondence should be addressed: Department of
Poultry Science, University of Georgia, Athens, GA 30602-2772.
375
na birds were more efficient at high SAA levels. Feather
weight gain increased in a linear manner with increasing
dietary SAA and was greater in na/na than Na/na birds
with high dietary SAA concentrations. Abdominal fat
decreased with increasing dietary SAA; and although
the Na/na birds had significantly more abdominal fat
than their na/na siblings (P = 0.049), on average the
difference was small and complicated by differences in
body weight. Analysis of covariance showed (a signifi-
cant interaction) that the relationship between abdomi-
nal fat and body weight was different for the na/na and
Na/na chickens. Although the shape of the response
curves of na/na and Na/na chickens to dietary SAA are
different, the quantitative requirements are very similar
during the growing period.
1996 Poultry Science 75:375-380
quirements of chickens, although they found few protein
level by genotype interactions for fast- and slow-
feathering broilers. Effects of feathering on specific
amino acid requirements have not been investigated.
Like the FL chickens, naked neck chickens have a
reduced proportion of feathers; but unlike the FL birds,
naked neck chickens have an increased proportion of
breast meat yield at normal and high temperatures
(Zein-El-Dein et al, 1984; El Attar and Merat, 1985;
Cahaner et al, 1993).
We hypothesized that the naked neck chickens have
reduced SAA requirements because of their decreased
amount of feather synthesis (Na/Na have approximately
30% less feathers than na/na; Bordas et al, 1978). The
growing period, 28 to 42 d, was studied because it is
when broilers eat the largest amount of feed. The
experiment described here used conditions practically
identical to those of Leclercq et al. (1993), except that the
genetic stocks were the result of birds with the naked
neck gene being crossed into the LL to produce full
brothers carrying either Na/na (naked neck) or na/na
(normally feathered) genes.
MATERIALS AND METHODS
The birds came from the cross of a homozygous
female line (na/na; the LL described by Leclercq, 1988)
376 PESTI ET AL.
with a heterozygous Na/na male line. The N a / n a male TABLE 1. Composition of the basal diet
line was produced by crossing LL females (Leclercq,
Ingredients Basal diet
1988) with an Na/Na commercial grandparent line. 3
Chickens were fed a common starter diet from (g/kg)
hatching to 28 d of age when the experimental period Ground yellow corn 200
Soybean meal 300
started. Male birds (14 per treatment) were placed in Cornstarch 403.63
individual cages in a temperature-controlled room (22 C Rapeseed oil 50
constant). They were offered one of the five experimen- Dicalcium phosphate 20
Calcium carbonate 10
tal diets made by adding DL-methionine to the basal Vitamin premix 1 5
diet described by Leclercq et al. (1993). The basal diet Mineral premix 2 1
was based on corn, soybean meal, cornstarch, rapeseed Sodium chloride 4
L-lysine hydrochloride 2.2
oil, and synthetic L-amino acids (Table 1). The basal diet L-threonine 0.95
contained 5.4 g SAA/kg 2 and 161 g crude protein/kg. 4 L-histidine 0.70
Diets were provided as pellets (2.5 mm diameter) for L-tryptophan 0.12
L-valine 0.80
ad libitum consumption. At the beginning of the L-leucine 1.10
experimental period (28 d of age) five birds per L-arginine hydrochloride 0.50
genotype were killed after an overnight period without Composition by calculation
feed to determine body composition (carcass protein and (NRC, 1994)
Protein 1.61
lipids) and feather weight. ME n , kcal/g 3.20
The experimental period lasted 16 d (28 to 42 d of SAA 5.4
age). The evening when the birds were 41 d old, feed lr
Trace mineral mix provides (in milligrams per kilogram of diet):
was withdrawn and the birds were weighed on the Mn (manganese oxide), 70; Zn (zinc oxide), 50; Fe (ferrous sulfate), 70;
morning of their 42nd d. After weighing, all chickens Cu (copper sulfate), 6; Co (cobalt carbonate), 0.3; I (potassium iodide),
1.1; calcium carbonate, 803.
were killed by saturated KC1 injection. Ten randomly 2
Vitamin premix provides (per kilogram of diet): vitamin A (retinyl
chosen birds per treatment were dry plucked just after acetate), 12,000 IU; cholecalciferol, 200 IU; vitamin E (dl-a-tocopheryl
death. Feather weight was determined by weighing acetate), 34 IU; riboflavin, 4 mg; calcium pantothenate, 8 mg; nicotinic
birds before and after plucking. After the birds were acid, 15 mg; choline CI, 190 mg; vitamin B6, 1 mg; menadione (as
menadione sodium bisulfite), 1 mg; thiamin, 1 mg.
chilled, abdominal fat pads were removed and weighed.
Blood samples were collected at 40 d of age from
birds fed 5.4 and 7.0 g SAA/kg. For plasma amino acid
analysis, plasma protein was precipitated by sulfosali-
cylic acid (Geraert et al, 1987). Plasma amino acid RESULTS
concentrations were then determined after ion exchange At 28 d the na/na birds weighed 656 ± 70 g (mean ±
chromatography.
standard deviation) with 5.91 ± 0.58% feathers, and the
Statistical analyses were performed using the GLM
Na/na birds weighed 673 + 61 g with 5.45 ± 0.56%
and NLIN procedures of SAS® (SAS Institute, 1985). The
feathers. The difference in total body weight was not
experimental model for the analysis of variance included
significant at P > 0.10, but the difference in feathers was
terms for the linear and quadratic effects of SAA level,
significant at P < 0.001.
genotype, and an SAA level by genotype interaction,
and block effects for side and tier of the cage unit. The The performance of the groups is presented in Table
experimental model for the analysis of covariance 2. As the dietary SAA content increased from 5.4 to 7.0
included terms for SAA level, genotype, gain, SAA level g/kg, body weight gain and feed consumption increased
by genotype interaction, and gain by genotype interac- in a curvilinear manner. There were no significant
tion. The SAA requirements for maximum performance differences between genotypes or SAA by genotype
were estimated with a model with an ascending interactions detected by analysis of variance (P < 0.05),
quadratic curve and plateau as follows: although the F test indicated that there was only about
one chance in eight that the difference in body weight
PROC NLIN; PARMS MAX=n RC=-205 REQ=6; gains of the genotypes was due to chance alone (P =
1=0; IF SAA<REQ THEN 1=1; 0.125). Feed efficiency (grams of gaimgrams consumed)
MODEL Response=MAX+RC*((REQ-SAA)**2)*I; increased curvilinearly with increasing dietary SAA.
Overall there was very little difference in the feed
where "MAX=n" is the maximum response (n = 670 for efficiency of Na/na vs na/na birds, but the Na/na birds
gain, and 0.43 for feed efficiency); "RC" = the rate were more efficient at low SAA levels, and na/na birds
constant; "SAA" = the SAA content of the diet in grams were more efficient at high SAA levels, accounting for
per kilogram; and "REQ" = the SAA requirement. the significant SAA by genotype interaction (Table 3).
Feather weight gain increased in a linear manner with
increasing dietary SAA (P < 0.05) and was greater in na/
3
SASSO, Sabres, France. na than in Na/na birds, as evidenced by the significant
4
Determined by Eurolysine, 80084 Amiens, France. interaction of genotype and SAA (P < 0.05). This
 SULFUR A M I N O ACIDS FOR N A K E D N E C K S 377
TABLE 2. Body weight gain, feed consumption, feed efficiency, and body composition during the experimental period

Dietary
methionine Weight Feed Feed Feather Abdominal
Genetic stock + cystine Parameter gain consumption efficiency gain fat

(gAg) M - (g=g) -(g)

Naked neck 5.4 5c 603
«/1,571 44 29.3
0.383
SD 80 146 0.023 6 9.0
5.8 5< 662 1,598 0.414 43 30.0
SD 56 116 0.019 8 5.9
6.2 5c 672 1,624 0.414 48 33.2
SD 89 173 0.031 12 10.7
6.6 X 681 1,569 0.434 46 26.5
SD 83 156 0.031 7 8.2
7.0 5c 645 1,507 0.428 45 20.9
SD 68 112 0.028 7 7.2
Normal 5.4 X 564 1,582 0.357 41 24.0
SD 74 188 0.031 6 6.8
5.8 X 644 1,622 0.396 45 30.2
SD 91 105 0.042 12 7.5
6.2 x" 667 1,561 0.431 45 24.7
SD 127 129 0.094 11 7.9
6.6 X 674 1,552 0.434 53 25.7
SD 97 117 0.050 9 5.0
7.0 X 678 1,519 0.446 55 26.2
SD 89 147 0.031 10 5.7
Naked neck X 652 1,576 0.414 45 27.9
SD 80 145 0.032 8 9.1
Normal 3< 645 1,567 0.412 48 26.1
SD 103 140 0.062 11 6.8
Analysis of variance df — Probabilities —
SAA 1 <0.001 0.050 <0.001 0.002 0.046
SAA x SAA 1 0.004 0.058 0.027 0.793 0.039
Genetic stock 1 0.125 0.862 0.037 0.033 0.049
SAA x genetic stock 1 0.137 0.844 0.040 0.023 0.062
Coefficient of determination 0.203 0.126 0.322 0.170 0.129

difference was much more pronounced at high dietary abdominal fat at low dietary SAA concentrations, and
SAA levels (Table 3). The difference between genotypes less abdominal fat at high dietary SAA concentrations,
was related to changes in total body weight gains an overall pattern was not clear. Analysis of covariance
because feather gain as a proportion of total gains (Table 3) indicated that there were differences in the
remained fairly constant. The Na/na birds had signifi- relationships between body weight gain and both feed
cantly more abdominal fat than the na/na birds. consumption and abdominal fat pad weights. At higher
Although the SAA by genotype interaction was nearly body weights, naked neck chickens ate more feed and
significant and Na/na birds tended to have more deposited the additional energy in abdominal fat than
their normal brothers (Figure 1).
The SAA requirements for gain, as estimated by
nonlinear regression, were 6.24 ± 0.45 (R2 = 0.171) and
55 5.96 + 0.52 g / k g (R2 = 0.107) for the na/na and Na/na
50 genotypes, respectively. The SAA requirements for feed
45 efficiency were 6.87 ± 0.56 (R2 = 0.280) and 6.80 ± 0.53 g /
H 40 kg (R2 = 0.291) for the na/na and Na/na genotypes,
< respectively. There were significant differences in
>* 3 5
plasma amino acid concentrations for 10 of the 17 amino
a 3° acids measured due to genetic stock (Figure 2), but only
1 25
2 5 due to methionine supplementation (Figure 3; Table 4).
% °
15
10 DISCUSSION
300 400 500 600 700 800 900 1000 1100 It is clear from these results that the heterozygous
naked neck chickens responded differently to SAA
BODY WEIGHT GAIN (g) concentration than homozygous normally feathered
siblings. The naked neck chickens performed better than
FIGURE 1. The abdominal fat pad weight by body weight gain
relationships of normal (na/na; solid line, filled circles) and naked neck
their siblings at low dietary SAA concentrations, espe-
(Na/na; broken line, open circles) chickens. cially for feed efficiency and growth. Because the naked
378 PESTI ET AL.
TABLE 3. Results of analysis of covariance of data from Table 1 (Type III sum of squares)

Feed Feather Abdominal

Analysis of covariance consumption gain fat
Probabilities
SAA 0.005 0.026 0.002
Genetic stock (GS) 0.689 0.083 0.875
SAA x GS 0.693 0.151 0.188
Gain <0.001 0.004 <0.001
Gain x GS •cO.001 0.403 0.019
Coefficient of determination (R2) 0.575 0.252 0.319

neck birds gained more total weight at low SAA
concentrations, they actually had more feather weight
gain than their normally feathered siblings, despite a
lower percentage of total gain as feathers.
The differences in weight gain and feed efficiency
responses corresponded to only very small differences in
requirements for maximum performance. The differ-
ences in SAA requirements of 0.28 g / k g for gain and
0.07 g / k g for feed efficiency are well within the error of
an experiment of this size. The problem of measuring
requirements very precisely with an experiment of this
size comes from the relatively small overall response to
SAA (overall about 114 g), coupled with the high
individual variability (standard deviations averaged 65
g). The individual data used here, as opposed to pen
averages in many other experiments, makes the R 2
values appear small. Individual variation is usually
ignored in models that only consider pen averaged data
so the R2 values do not reflect the total variation in the
experiment (including between birds).
Although there may be large differences in response
at low SAA levels, the response lines converge near the
requirement, making the point at which the plateau is
reached very difficult to pinpoint. The inclusion of even
a small margin of safety above the SAA requirement of
the naked neck chickens would include the estimate for
the normally feathered chickens. Therefore, from a
practical standpoint, the SAA requirements of na/na and
Na/na birds can be considered to be the same.
The naked neck birds had significantly less (P < 0.001)
feather weight at 28 d, although the actual difference in
feather weight gain during the subsequent 2-wk period
studied here was very small. Any difference in gain was
essentially nonexistent at suboptimal SAA levels, but
was about 10 g with excesses of SAA. Birds without
adequate SAA could not preferentially produce smaller
feathers while maintaining other lean tissue growth. If
the supply of SAA was not sufficient to allow for
maximum body weight growth, feather growth was
decreased slightly. Increasing the SAA above the level
necessary to result in maximum growth (> 6.2 g/kg)
resulted in a large increase in feather growth but no
further reduction in abdominal fat pad size in the
normally feathered birds. In contrast, for the naked neck
birds (Na/na), increasing the amount of SAA above the
requirement for maximum growth resulted in no
increase in feather gain, but a decrease in abdominal fat
pad size.
The data reported herein challenge field observations
that birds fed diets low in methionine suffer from poor
feathering. The very deficient naked neck birds actually
had a higher proportion of feathers at low SAA levels,
but then total body growth increased faster than feather
growth as SAA levels increased. Near the SAA require-

16 T 20

u
4
a 16
4 12
z S"
P 10

o
z
o ez
o o
u

II Itirlff d I I
" " WW

3
El,
f s 88
a. J •" <

FIGURE 2. The plasma amino acid concentrations of chickens fed FIGURE 3. The plasma amino acid concentrations of naked neck
5.4 g/kg (black bars) or 7 g/kg (open bars) sulfur containing amino chickens {Na/na; black bars) or their normally feathered brothers (na/
acids. Asterisks indicate significant effect of diet at P < 0.05 (*) or P < na; open bars). Asterisks indicate significant effect at P < 0.05 (*) or P <
0.01 (»*). 0.01 (»*).
 SULFUR AMINO ACIDS FOR NAKED NECKS 379
ment, the normally feathered birds had a higher
proportion of feathers. Above the SAA requirement,
feather and total growth continued to increase for the
o o o o
normal birds, but decreased for the naked neck birds.
4> l-H t ^ i-l The changes resulted in the sigmoid shaped responses
CO I - I ^ H I T ) ( N I
C O O C N O C O O C S O
^ S S <N observed.
o ©o o
IT) n T}< CO
The results of the analysis of covariance (Table 3) are
n Q r t f N
O O O \D particularly interesting because of the significant interac-
rtHoddcorlcod
tions found between body weight gain and both feed
consumption and abdominal fat. This analysis indicates
sO ON 0 0 O
that of naked neck and normal siblings of the same
1—I O i — I O T — < T-H I — l O d o o o above average weight, the naked neck one would be
rt o a> • *
expected to eat more feed and deposit that feed as
CM i-H t ^ l-H
i n r-^ i n o abdominal fat. For birds below average weight, the
( D r - i r O O C O O f O O o d d o normally feathered brother would have more abdominal
N O I O H
fat (Figure 1). Clearly the Na gene does affect feed
o o u i i n
a s o c o CN consumption, but the difference may be obscure unless
C O O C N O C O O f N O
d ddd
the gain by genetic stock probability is calculated.
O N p T t j p o q p i n p
CM i-H « **
Although analysis of variance indicates variation in
U > O H 4
d ddd
V
feather gain was due to SAA level and an SAA level by
stock interaction (P = 0.033), the interaction was n o
O H t O N
i n o i n i-i
CO O O C"^
longer significant when differences in body weight gain
d dd d were accounted for (P > 0.08; analysis of covariance,
V

Table 3). The biggest factor determining how much the

™ © o i n co birds ate, their feathering, and increases in fat pad
' O O O O O •g o d
^ V
d d
weight, was their amount of body weight gain.
Tfl T-I Ov • *
C J O i C H
The results of the plasma amino analyses were very
( N O r t H O q i f l r H a> o I-H i n similar to what was observed by Leclercq et al. (1993),
who fed similar diets to FL and LL chickens (Table 4;
Figure 2). When methionine was supplemented in the
diet, the level of plasma methionine rose, but most of
I N O K O 0 0 O 0 0 O o o o o
the other amino acid concentrations were significantly
CO i-H CO O
oo © i n • # reduced. These results are typical of amino acid
i—i a ON sO
^ O O t O N O l f t O d ddd supplementation (Zimmerman and Scott, 1965, 1967;
Peng and Harper, 1970). The exceptions were the
. if) q c o o \ q
N N H O \
m H n r t
^ O i O O O
aromatic amino acids, aspartic and glutamic acids, and
i O r-5 O O <-< o d d © alanine, which were essentially unchanged.
The results presented in Tables 2 and 3 and Figures 1
K r i H l / ) 0 \ O t S \ D
o o co »-i
CM CO K rH
CM H "tf i-H
and 2 demonstrate that although there are demonstrable
N O c N O N O r o d d d d d differences in the growth, fattening and plasma amino
CM i-H L0 O acid concentrations of heterozygous naked neck chick-
t-~ Q HH O

O O t N . O ( N O t N O
o o n s ens vs normal chickens, these differences were not large
enough to result in differences in the stock's quantitative
CM i-H CM CM
O Q vO K
requirements. This result may be largely d u e to the
t o vq ^ ro oq t o oq '
^ d c o d o 6 d o \ o
O © -^ 00
d dd d period that was studied. There were no large differences
V
in feather growth between the stocks from 4 to 6 wk of
N O* t H age. It may be that during the first 4 wk of life the large
N C O O f O N T t i N i n
L D O ^ O O ^ O O ^ O O
O J i l O H
^ N O O O differences in feather growth result in measurable
o o o o o o o o d ddd
differences in SAA requirements.

Q Q D Q UH
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