Ganadería Ciencia 168 (2014) 94 - 101

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Efecto de la reducción del contenido de proteína cruda de dietas suplementadas con

aminoácidos esenciales en el rendimiento de los lechones con un peso 6 - 15 kg

JB Toledo un , norte , AC Furlan un , PC Pozza un , LM Piano segundo , OLP Carvalho do ,

LM-Peñuela Sierra re , LMD Huepa un
un Departamento de Zootecnia, Universidad Estatal de Maringá, Maringá, Brasil
segundo Universidade Federal do Mato Grosso do Sul, Campo Grande, Brasil
do Universidad Federal de Bahía, Salvador, Brasil
re Universidad Cooperativa de Colombia, Neiva, Colombia

información del artículo abstracto

Historia del artículo: Se realizaron dos experimentos para evaluar el efecto de la reducción del contenido de proteína bruta (CP) de dietas
Recibido el 7 de agosto de 2013 recibido suplementadas con aminoácidos en lechones de un peso 6 - 15 kg. En el experimento de rendimiento (Experimento I), 120
en forma revisada el 10 de julio de 2014
lechones destetados a los 21 días de edad con pesos vivos iniciales de 5,95 7 0,33 kg se distribuyeron en cinco grupos de
tratamiento. Esta agrupación siguió un diseño de bloques al azar con ocho repeticiones y tres animales por unidad
Aceptado 13 de julio de 2014
experimental. Los tratamientos consistieron en cinco dietas diferentes, en los que el contenido de CP se redujeron de
21,0% a 15,0% (21,0%, 19,5%, 18,0%, 16,5% y 15,0% CP); los requisitos de aminoácidos de la dieta fueron recibidos por
la adición de L-lisina, DL-metionina, L-treonina, L-triptófano, L-valina y L-isoleucina. No se encontraron diferencias en las
variables asociadas con el rendimiento entre los animales de diferentes grupos de tratamiento. Por lo tanto, cualquiera de
palabras clave:
los niveles de PC investigados puede utilizarse eficazmente en las dietas de los lechones suplementados con aminoácidos
La proteína cruda
nitrógeno proteico sintéticos. La relación esencial / no esencial aminoácido (EAA: NEAA) aumentó con la reducción del contenido de CP, y la
Ideal urea porcina mejor relación (53: 47) se logró con la dieta que contenía 15% de proteína. concentraciones de urea disminuyeron
linealmente con la reducción de la proteína (Experimento I). Para evaluar el balance de nitrógeno (Experimento II), 20
lechones machos mestizos castrados de un linaje comercial, destetados a los 21 días de edad, fueron asignados al azar en
dos bloques, en los que cada bloque tenía dos repeticiones (cuatro repeticiones por tratamiento). El peso vivo promedio de
los lechones fue 10,79 7 2.19 kg. Los animales se alojaron en jaulas de metal y se distribuyeron en cinco grupos de
tratamiento siguientes un diseño de bloques al azar con cuatro repeticiones; la unidad experimental consistió en un lechón.
La excreción de nitrógeno y las concentraciones de urea en sangre y de orina disminuyeron linealmente ( PAG o 0,05) con
la reducción de la CP en las dietas, lo que resulta en una reducción de la excreción de nitrógeno en el medio ambiente.

y 2014 Elsevier Todos los derechos reservados.

abreviaturas: EAA, aminoácido esencial; De NEEA, amino no esencial ácido

1. Introducción
norte Correspondencia a: Departamento de Zootecnia, Universidad Estatal de Maringá,

Maringá, PR 87020-900, Brasil. Tel .: þ 55 44 3268 5813.

Los contaminantes excretados por los cerdos constituyen uno de los principales

Dirección de correo electrónico: juliana.b.toledo@gmail.com (JB Toledo). problemas asociados a la ganadería porcina moderna.

http://dx.doi.org/10.1016/j.livsci.2014.07.006
1871-1413 / y 2014 Elsevier Todos los derechos reservados.
 JB Toledo et al. / Ganado Ciencia 168 (2014) 94 - 101
Para mitigar los efectos de estos contaminantes, los investigadores han desarrollado las
dietas basadas en la noción de contenidos de proteína ideal.
La aplicación del concepto de proteína ideal a través de la suplementación con
cantidades apropiadas de aminoácidos sintéticos permite la reducción de la proteína
cruda de contenidos (CP) de dietas de los cerdos. Cole y Van Lunen (1994)
sugirió que cuando las dietas se formulan en base a la noción de contenido de proteína
ideal, debe lograrse una mezcla perfectamente equilibrada de aminoácidos esenciales
(EAA) para asegurar suficiente de nitrógeno para la síntesis de aminoácidos no
esenciales (NEAA). Sin embargo, a pesar de que la NEAA cuantitativamente representan
una fracción importante de proteína consumida ( Heger, 2003 ), Se ha prestado poca
atención al establecimiento de requisitos AANE oa la relación de EAA / AANE ideal para
cerdos.
La literatura no es concluyente sobre la composición de la dieta ideal para lechones
destetados. Los niveles de harina de soja incluidos en la dieta han sido discutidos, ya
que este componente en particular contiene factores alergénicos y anti-nutricionales que
reducen el índice de rendimiento de los lechones recién destetados (Zangeronimo,
2006). Como resultado, otras fuentes de proteínas en las dietas de los lechones han sido
estudiados, así como la inclusión de aminoácidos sintéticos.
El objetivo de la utilización de aminoácidos sintéticos es aumentar la eficiencia de
utilización de la proteína mediante la maximización de la utilización de los aminoácidos
en la síntesis de proteínas y la minimización de su uso en la producción de energía.
Además, el uso de aminoácidos sintéticos puede reducir la contaminación ambiental
causada por la excreción excesiva de nitrógeno.
Las dietas con altas concentraciones de CP o proporciones de aminoácidos
desequilibradas aumentan cerdos ' la ingesta de agua y, en consecuencia, dan como
resultado un aumento del volumen de la suspensión y una mayor excreción de nitrógeno ( Perdomo repeticiones por tratamiento). El peso vivo promedio de los lechones fue 10,79 7 2.19 kg.
y Lima, 1998 ). En teoría, las pérdidas de nitrógeno debido a los desequilibrios de
aminoácidos podrían reducirse o corregirse mediante la adición de aminoácidos sintéticos a
la dieta.
El objetivo de este estudio fue evaluar el rendimiento, el balance de nitrógeno, y la
concentración de proteínas y de sangre y orina metabolitos en los lechones con un peso
6 - 15 kg alimentados con dietas bajas en proteínas suplementadas con aminoácidos
sintéticos.
2. Materiales y métodos
Se realizaron dos experimentos con lechones de un peso 6 - 15 kg. El primer
experimento evaluó su rendimiento (Experimento I) y el segundo su equilibrio de
nitrógeno. Los experimentos se llevaron a cabo de acuerdo con las recomendaciones del
Comité para el comportamiento ético en el uso de animales en la experimentación de la
Universidad Estatal de Maringá.
2.1. Tratos
Los tratamientos consistieron en cinco dietas diferentes en la cual el contenido de
PC se redujo sucesivamente por 1,5% (21,0%,
19,5%, 18,0%, 16,5% y 15,0%), lo que resulta en las dietas bajas en proteínas. Las
dietas se formularon con base en el concepto de proteína ideal y siguiendo las
recomendaciones de
Rostagno et al. (2005) ; la aplicación del coeficiente de
95
de digestibilidad real se utilizó para estimar la cantidad de aminoácidos digestibles en la
dieta ( tabla 1 ). El contenido de aminoácidos de las dietas se calculó basándose en la
composición analizada de los ingredientes que contienen proteínas. Los siguientes
aminoácidos se consideraron como elementos esenciales: lisina, metionina, treonina,
triptófano, valina, isoleucina, arginina, leucina, fenilalanina e histidina.
2.2. Animales y diseño experimental
Para evaluar el rendimiento de los animales (Experimento I), se utilizaron 120
lechones mestizas de un linaje comercial. Los animales (60 machos y 60 hembras)
fueron destetados a los 21 días de edad y tenían un peso inicial promedio de 5,95 7 0,33
kg y un peso final promedio de 16,17 7 1,52 kg. Los animales se alojaron en un edificio
vivero que contiene plumas que proporcionaron 1.32m 2 de la superficie total, con suelos
de plástico parcialmente enrejado y un bebedor mordedura pezón en la zona trasera.
Alimentos y agua fueron suministrados libremente durante el experimento.
Los animales se distribuyeron en cinco grupos de tratamiento siguiendo un diseño
de bloques al azar con ocho repeticiones por tratamiento y tres animales por unidad
experimental (UE). Los animales se pesaron al inicio y al final del experimento, y la
ingesta total de alimento se registró. Los datos se utilizaron para calcular la ingesta diaria
de alimentación (CMD), ganancia diaria de peso (DWG), y alimentar a la tasa de
conversión (FCR) por la UE.
Para evaluar el balance de nitrógeno (Experimento II), 20 lechones machos
mestizos castrados de un linaje comercial, destetados a los 21 días de edad, fueron
asignados al azar en dos bloques, en los que cada bloque tenía dos repeticiones (cuatro
Los animales fueron alojados en jaulas metabólicas en una habitación en
condiciones parcialmente controladas. La temperatura mínima y máxima media
registrada durante el experimento fue 23,40 7 0,56 y 26,45 7 0.17 1 C, respectivamente.
Los animales se distribuyeron en cinco grupos de tratamiento siguientes un diseño de
bloques al azar con cuatro repeticiones por tratamiento; un lechón representó a la UE.
Las heces y la orina se recogieron de acuerdo con métodos descritos por Sakomura y
Rostagno (2007) . Los lechones fueron alimentados con cuatro comidas por día a las
7:30, 10:00, 13:30, y 16:30 h, que corresponde a 38%, 19%, 19% y 24% de la cantidad
total de alimento, respectivamente. La ingesta total de alimento diario se calculó de
acuerdo a la ingesta durante la fase de adaptación y basado en el peso metabólico del
animal (kg 0,75). Para evitar las pérdidas y para facilitar la manipulación, la comida se
humedece con 20% de su masa de agua. Para no perjudicar a los animales ' alimentar la
ingesta debido a la ingesta excesiva de agua, el agua se suministra después de cada
comida en una proporción de 3 ml / g de pienso.
Para calcular el balance de nitrógeno, una colección total de heces se realizó para
los que el óxido férrico 2% (Fe 2 O 3) se añadió a las dietas para marcar el comienzo y el
final de la recogida de las heces. Todo el material fecal producida por los animales se
recogió en bolsas de plástico una vez por día y se almacena en un congelador a
18 1 C. Para medir el contenido de nitrógeno, la
heces se homogeneizaron y se tomó una muestra de 20%.
96 J.B. Toledo et al. / Livestock Science 168 (2014) 94 – 101

tabla 1
Composición (g / kg) y el valor nutricional de las dietas que contienen diferentes niveles de proteína cruda para lechones de 6 a 15 kg.

ingredientes Nivel de proteína cruda,%

21.00 19.50 18.00 16.50 15.00

Maíz 428.00 473,11 519,98 567,43 614,41

harina de soja, 45% 275.90 228,36 178.51 127,90 77.18
Leche desnatada 150.00 150.00 150.00 150.00 150.00
Suero en polvo 70.00 70.00 70.00 70.00 70.00
Azúcar 30.00 30.00 30.00 30.00 30.00
Aceite de soja 13,91 12.18 9.82 7.22 4.50
El fosfato dicálcico 11.70 11.97 12.25 12.54 12.83
Caliza 6.24 6.45 6.67 6.89 7.11
Mineral - premezcla de vitaminas un 5.00 5.00 5.00 5.00 5.00
sal 5.52 3.95 2.39 0.82 -
Bicarbonato de sodio - 2.30 4.60 6.90 9.20
L-lisina HCl (78%) 1.02 2.42 3.88 5.37 6.85
DL-methionine (98%) 1.61 2.00 2.42 2.84 3.26
L-threonine (98%) 0.89 1.55 2.24 2.94 3.64
L-valine (96,5%) – 0.52 1.31 2.12 2.92
DL-tryptophan (98%) – – 0.26 0.53 0.80
L-isoleucine (98%) – – 0.49 1.32 2.14
Antioxidant – BHT 0.10 0.10 0.10 0.10 0.10
Leucomag s b 0.10 0.10 0.10 0.10 0.10

Calculated values EB
(mEq/kg) 232.9 232.7 231.2 229.4 227.7
ME (MJ/kg) 33.25 33.25 33.25 33.25 33.25
Lactose (g/kg) 131.1 131.1 131.1 131.1 131.1
Crude protein (g/kg) 210.0 195.0 180.0 165.0 150.0
Digestible lysine (g/kg) 13.30 13.30 13.30 13.30 13.30
(100) (100) (100) (100) (100)
Digestible methionine (g/kg) 4.97 5.17 5.38 5.59 5.80
(37.37) (38.87) (40.45) (42.03) (43.61)
Digestible threonine (g/kg) 8.38 8.38 8.38 8.38 8.38
(63.01) (63.01) (63.01) (63.01) (63.01)
Digestible tryptophan (g/kg) 2.50 2.26 2.26 2.26 2.26
(18.80) (16.99) (16.99) (16.99) (16.99)
Digestible valine (g/kg) 9.40 9.18 9.18 9.18 9.18
(70.68) (69.02) (69.02) (69.02) (69.02)
Digestible isoleucine (g/kg) 8.38 7.63 7.32 7.32 7.32
(63.01) (57.37) (55.04) (55.04) (55.04)
Digestible arginine (g/kg) 11.91 10.55 9.11 7.64 6.17
(89.55) (79.32) (68.50) (57.44) (46.39)
Digestible leucine (g/kg) 17.24 16.14 14.99 13.82 12.65
(129.62) (121.35) (112.71) (103.91) (95.11)
Digestible phenylalanine (g/kg) 9.53 8.70 7.82 6.93 6.04
(71.65) (64.51) (58.80) (52.11) (45.41)
Digestible histidine (g/kg) 5.15 4.75 4.33 3.91 3.49
(38.72) (35.71) (32.56) (29.40) (26.24)
Calcium (g/kg) 8.25 8.25 8.25 8.25 8.25
Sodium (g/kg) 3.50 3.50 3.50 3.50 3.79
Chloride (g/kg) 6.12 5.46 4.81 4.16 3.96
Potassium (g/kg) 9.91 9.17 8.40 7.61 6.82
Available phosphorus (g/kg) 4.50 4.50 4.50 4.50 4.50
Ratio EAA:NEAA c 42:58 48:52 49:51 51:49 53:47

a Vitamin and mineral mix for piglets in the nursery phase, composition per kg of the product: vit. A – 1,800,000 IU; vit. D3 – 360,000 IU; vit. E – 4000 IU; vit. K3 – 600 mg; vit. B1 – 280 mg; vit. B2 – 800 mg; vit. B6 – 300

mg; vit. B12 – 3600 mg; niacin – 6000 mg; pantothenic acid – 3200 mg; biotin –
20 mg; folic acid – 80 mg; choline – 31 g; Fe – 20,000 mg; Cu – 50,000 mg; Co – 120 mg; Mn – 11,000 mg; Zn – 18,000 mg; Se – 60 mg; I – 200 mg; antioxidant – 20 g; vehicle Q.S.P. – 1000 g.

b Leucomag:leucomycin at 30%; EB – electrolyte balance; ME – metabolizable energy.

c Ratio EAA:NEAA – Ratio essential amino acid:nonessential amino acid; The amino acid values between parentheses correspond to ideal protein values. The digestible amino acid values were obtained from Rostagno

et al. (2005) .

Se recogió la orina en cubetas de plástico que contienen 20 ml de HCl 1: 1, de la 2.3. Análisis químico
que se tomó una alícuota de 20% diario y se almacenaron a
18 1 C. La orina acumulada fue homo- El contenido de aminoácidos del maíz, harina de soja, leche desnatada en polvo, y
nizado, y se tomaron muestras para medir los niveles de nitrógeno, urea, y creatinina. suero de leche en polvo se evaluaron mediante cromatografía líquida de alta resolución
 J.B. Toledo et al. / Livestock Science 168 (2014) 94 – 101 97

(HPLC) by Ajinomoto of Brazil; the coefficients of ileal standardized digestibility of the
amino acids, described by
Rostagno et al. (2005) , were applied to the analyzed values to obtain the values of
digestible amino acids of these feed ingredients
The dry matter (DM) analysis of the feces was performed in a 55 and 105 1 C stove,
and for the ingredients and the feed it was performed in a 105 1 C stove. The DM was
obtained subtracting the initial weight by the final weight of the samples. The Kjeldahl
method was used to perform nitrogen analysis, in which the method is based on the
decomposition of organic matter by digesting the sample at 400 1 C with concentrated
sulfuric acid. The resulting nitrogen in acid solution is determined by steam distillation
drag, followed by titration with dilute acid ( AOAC, 1990 ).
obtained by multiplying the nitrogen content by the amount of feed intake, excreted feces,
and urine, respectively. These values served to calculate the amount of retained nitrogen
(RN ¼ NI
FNE UNE), the percentage
of RN relative to NI which corresponds to the net protein utilization (NPU ¼ 100 RN / NI),
and the percentage of RN relative to the apparently absorbed nitrogen, which
corresponds to the biological value of food protein (BVFP
¼ 100 RN / (NI FNE)), as described by Adeola (2001) .
All variables were calculated based on the metabolic weight of the animal (kg 0.75).

2.5. Statistical analysis

The values of the variables corresponding to performance, nitrogen balance, blood

Blood samples were collected from all animals, using heparinized tubes from the
cranial vena cava at the end of each experiment. The following parameters were
measured: urea, creatinine, and total protein levels in Experiment I and urea, creatinine,
total proteins, albumin, and globulin levels in Experiment II.
chemistry (experiments I and II), and urine tests (Experiment II) were subjected to
analysis of variance by means of the general linear model (GLM) of SAS (2007) according
to the following statistical model:

Y ij ¼ m þ B i þ b 1 CP i þ b 2 CP i 2 i þ LA þ e ij;

Blood samples were centrifuged (3000 rpm for 15 min), and plasma was transferred
to labeled Eppendorf tubes and stored in a freezer ( 18 1 C) until analysis. To perform
the analysis of blood and urine metabolites,
“ in vitro ” diagnostic kits by Gold Analisa (Belo Horizonte, Brazil) were used; globulin
concentrations were calculated as the difference between total protein and albumin. The
absorbance of each analyte was read on a spectrophotometer BIOPLUS s 2000.
Urea was quantified by an enzymatic colorimetric method with urease. Creatinine
was determined by reaction with picric acid. Albumin was determined by reaction with
bromocresol green. The total protein was performed by a biuret reaction, which result in a
chemical complex whose color intensity is directly proportional to its concentration in the
analyzed sample.
where Y ij ¼ dependent variable, m ¼ overall average, B i ¼
block effect ( i ¼ 1, 2, 3, 4, 5, 6, 7, and 8 in Experiment I; or i ¼ 1 and 2 in Experiment II), b
1¼ coefficient of linear regression as a function of the CP level, CP i ¼ CP level ( T 1 ¼ 21, T
2¼ 19.5, T 3 ¼ 18, T 4 ¼ 16.5, and T 5 ¼ 15%),
b 2 ¼ coefficient of quadratic regression as a function of the CP level, LA ¼ lack of
adjustment of the regression model, and e ij ¼ experimental error associated with
observation. The linear effect of the reduction of CP on the dependent variable was
established. Differences were considered to be significant when P o 0.05.
The initial weights of piglets were used as covariates of the variables ADFI, DWG,
and FCR.

3. Results

In Experiment I, the reduction of the dietary CP was possible by replacing the

2.4. Calculation of the Nitrogen Balance (Experiment II) soybean meal with corn grain. This reduction in CP did not produce significant differences
( P 4 0.05) in the performance of piglets weighing 6 – 15 kg ( Table 2 ).
The values of nitrogen intake (NI), fecal nitrogen excretion (FNE), and urine nitrogen
excretion (UNE) were

Table 2
The performance of pigs fed diets differing in crude protein content and supplemented with essential amino acids.

Item Level of crude protein, %

21.0 19.5 18.0 16.5 15.0 RSD e Effect f

IBW a , kg 6.09 5.92 5.90 5.92 5.94 0.06 NS g

FBW b , kg 16.394 16. 333 16.538 16.128 16.439 0.26 NS
ADFI c , g 535 525 506 498 538 0.01 NS
ADG d , g 348 347 354 340 349 0.01 NS
Feed:Gain 1.542 1.525 1.436 1.478 1.541 0.02 NS

a Initial body weight.

b Final body weight.
c daily feed intake.
d daily weight gain.
e Residual standard deviation.
f Regression analysis – polynomial regression analysis.
g Nonsignificant.
98 J.B. Toledo et al. / Livestock Science 168 (2014) 94 – 101

The serum urea levels declined linearly ( P o 0.05) with
decrease in protein content. No significant differences ( P 4 0.05) were found in the
creatinine and total protein levels among treatments ( Table 3 ).
In Experiment II, the NI, UNE, and apparently absorbed nitrogen measured based on
the metabolic weight (PV 0.75)
declined linearly ( P o 0.05) with the dietary CP content ( Table 4 ). The FNE did not vary
with dietary protein ( P 4 0.05).
The total nitrogen excretion decreased ( P o 0.05) as
protein content decreased, while the net protein utilization (NPU) and biological value of
the feed protein increased linearly ( P o 0.05). The nitrogen retention (g/kg PV 0.75/ day)
decreased linearly with dietary protein content ( P o 0.05). With the exception of urea
which declined linearly with protein content ( P o 0.05), no differences were found ( P 4 0.05)
in the levels of creatinine, total protein, and albumin among animals from different
treatment groups (Table 5). In the same way, the urine urea levels of the piglets
decreased linearly ( P o 0.05) with decreasing dietary CP content (Table 6) and, on the
other hand, the creatinine level increased linearly ( P o 0.05) according to the decrease in
the protein levels in the piglets ’ diets.
et al. (2006) showed that the DWG and FCR of piglets with initial average weights of 8.55
kg were not influenced by decreasing CP levels (down to 16.5%).
Earlier experiments, such as that of Lordelo et al. (2008) ,
which assessed the performance of piglets weighing 5 – 20 kg, indicate that it is possible to
reduce the CP of diets if these diets are supplemented with limiting amino acids.
According to Wu (1998) , the inclusion of synthetic amino
acids in diets might instigate competition among the amino acids for intestinal absorption
sites and sites of protein synthesis because synthetic amino acids are absorbed faster
than the amino acids found in food protein. However, in the present study, the
performance-related variables did not appear to be affected by this phenomenon.
In the present experiment, excess levels of histidine (21.0% and 19.5% of CP) were
found in certain treatments, while insufficient quantities were encountered in others.
However, no impairments in performance were observed, indicating that the histidine
requirements were most likely satisfied in all diets. Further studies are needed to
establish the histidine requirements of piglets.

4. Discussion
Similar to findings of this study, Kerr et al. (2003) did not observe any differences in
performance-related variables of piglets weighing 6 – 15 kg. Similarly, Zangeronimo
Rostagno et al. (2011) established the minimum levels of protein that must be
present in diets based on corn bran and soybean when EAA L-lysine, DL-methionine, and
L-threonine are provided.
Studies designed to provide better estimates of the EAA:NEAA ratio in rats, swine,
and poultry have shown that the best ratio for growth or protein deposition does not differ
substantially among species and varies from

Table 3
Level of biochemical variables in plasma and urine of pigs fed diets differing in crude protein content and supplemented with essential amino acids.

Item Level of crude protein, %

21.0 19.5 18.0 16.5 15.0 RSD a Effect b

Experiment I, plasma
Urea, mg/dL 26.3 23.9 20.5 15.4 12.4 0.74 Lc
Creatinine, 0.86 0.89 0.88 0.84 0.86 0.01 NS d
mg/dL
Total protein, 3.72 3.88 3.70 3.70 3.70 0.04 NS
g/dL
Regression equation R2
Urea ¼ 24.0625 þ 2.43492 CP 0.98

Experiment II, plasma

Urea, mg/dL 32.6 24.5 18.9 19.2 14.1 1.96 Lc
Creatinine, 0.96 0.92 0.99 0.91 0.99 0.02 NS d
mg/dL
Total protein, 3.84 3.72 3.55 3.79 3.66 0.07 NS
g/dL
Albumin, g/dL 1.96 1.76 1.69 1.98 1.56 0.07 NS
Globulin, g/dL 1.87 1.95 1.85 1.80 2.09 0.09 NS

Regression equation Rb
Urea ¼ 28.6497 þ 2.80917 CP 0.90
Experiment II, urine
Urea, mg/dL 505.3 412.6 319.8 227.1 134.3 2.2 Lc
Creatinine, 28.3 32.1 35.9 39.7 43.5 47.3 L
mg/dL
Regression equation Rb
Urea ¼ 793.331 þ 61.8427 CP 0.73
Creatinine ¼ 81.4563 – 2.52750 CP 0.71

a Residual standard deviation.

b Regression analysis – polynomial regression analysis.
c Linear effect.
d Non-significant.
 J.B. Toledo et al. / Livestock Science 168 (2014) 94 – 101 99

Table 4
The nitrogen balance of pigs fed diets differing in crude protein content and supplemented with essential amino acids.

Item Level of crude protein, %

21.0 19.5 18.0 16.5 15.0 RSD a Effect b

Nitrogen intake (g/day) 21.91 15.24 20.21 17.94 14.98 21.17 -

Nitrogen intake (g/kg BW 0.75/ day) 3.58 2.95 3.25 2.80 2.53 7.48 Lc
Fecal N (g/day) 2.46 2.10 2.27 2.09 2.23 21.91 -
Fecal N (g/kg BW 0.75/ day) 0.40 0.41 0.36 0.34 0.38 17.52 NS
Urinary N (g/day) 3.95 1.99 2.24 2.32 1.14 33.35 -
Urinary N (g/kg BW 0.75/ day) 0.64 0.38 0.35 0.36 0.19 21.75 L
N apparently absorbed (g/day) 19.45 13.15 17.94 15.85 12.75 21.85 -
N apparently absorbed (g/kg BW 0.75/ day) 3.18 2.54 2.90 2.46 2.14 7.89 L
Total N excretion (g/day) 6.41 4.08 4.51 4.41 3.37 25.60 -
Total N excretion (g/kg BW 0.75/ day) 1.05 0.79 0.71 0.70 0.57 15.17 L
N retained (g/day) 15.50 11.16 15.70 13.53 11.61 20.74 -
N retained (g/kg BW 0.75/ day) 2.54 2.16 2.55 2.10 1.95 7.61 L
N retained /N intake (%) – NPU d 70.83 73.27 78.44 75.01 77.29 4.12 L
N retained /N apparently absorbed (%) – BVFP e 79.87 85.03 87.94 85.27 91.25 3.04 L

Regression equation Rb
Nitrogen intake (g/kg BW 0.75/ day) ¼ 0.306790 þ 0.942604 N 0.76
Urinary N (g/kg BW 0.75/ day) ¼ 0.719801 þ 0.383643 N 0.80
N apparently absorbed (g/kg BW 0.75/ day) ¼ 0.0664506 þ 0.895048 N 0.72
Total N excretion (g/kg BW 0.75/ day) ¼ 0.479462 þ 0.431198 N 0.87
N retained (g/kg BW 0.75/ day) ¼ 0.786252 þ 0.511406 N 0.52
N retained /N intake (%) NPU ¼ 92.5520 – 6.10556 N 0.57
N retained /N apparently absorbed (%) BVFP ¼ 113.471 – 9.58355 N 0.75

a Residual standard deviation.

b Regression analysis - polynomial regression analysis.
c Linear effect.
d Net protein utilization.
e Biological value of the food protein. The data are from pigs with average body weight of 10.79 7 2.19 kg.

55:45 to 60:40 ( Heger, 2003 ). In the present study, the best EAA:NEAA ratio was 53:47
and was achieved with the diet consisting of 15% CP.

As expected, and as found by other authors ( Figueroa et al., 2002 ; Kerr et al., 2003 ),
plasma urea levels declined with decreasing levels of dietary protein. Serum urea levels
increase due to the excess catabolism of amino acids by deamination. Thus, when the
animal consumes appropriate qualities and quantities of protein, a better amino acid
balance is achieved in the diet; consequently, greater synthesis of protein and less amino
acid catabolism occur, leading to reductions in blood urea levels ( Nelson and Cox, 2011 ).

According to Kaneko et al. (1997) , urea levels from 10 to

30 mg/dL and creatinine levels from 1.0 to 2.7 mg/dL in blood are considered within
Fig. 1. Nitrogen excretion in the urine of piglets 6 – 15 kg according on the levels of crude protein.
normal limits in swine; the levels encountered in the present study fall within the normal
physiological ranges.

The lack of statistically significant differences in the creatinine and total protein levels
among animals of different treatment groups might be attributed to the fact that creatinine
is neither reabsorbed nor reutilized by the animal and therefore must be continually
removed through the kidneys; NI, conversely, seems to exert little effect on creatinine and
protein blood concentrations ( Nelson and Cox, 2011 ).
Shriver et al. (2003) reduced the dietary CP content by 4% and supplemented the
diets of growing swine with the same amino acids used in the present study; these
authors also encountered reduced levels of NI and absorption
(Experiment II). Based on these results, it can be assumed that the difference in
absorption rates of synthetic and feed-derived amino acids might contribute to different
passage rates of these nutrients from the gastrointestinal tract into the blood.
Zangeronimo et al. (2006) reduced the protein level in diets for pigs to 16.5% and
also found that the nitrogen excretion in the urine decreased. This reduction in nitrogen
from urine can also be explained by the fact that urine is the major route of elimination of
excess nitrogen; as a result, when the CP levels are reduced, environmental pollution
from pig manure may decrease. In the present
100 J.B. Toledo et al. / Livestock Science 168 (2014) 94 – 101
study, the nitrogen excretion in the urine was reduced by approximately 12%.
According to the regression equation adjusted for UNE ( 0.719801 þ 0.383643 X), the
UNE increased 0.3 g/day for every 1% increase in dietary nitrogen levels. Therefore, the
higher the nitrogen level of the diet, the higher the UNE will be ( Fig. 1 ).
The diets with the highest CP contents proved to be the most efficient, as the
nitrogen retention decreased linearly with dietary protein levels (g/kg PV 0.75/ day). These
findings were unexpected because the diets were formulated based on the ideal protein;
similar observations were made by Oliveira et al. (2005) . Other factors might have limited
the nitrogen retention. Heger et al. (2003) emphasize that the greater nitrogen retention
found in shortterm experiments might simply be due to adaptive responses to excessive
intake of nonessential nitrogen.
Another potential explanation for the reduction in nitrogen retention in diets with low
CP content can be attributed to the possible overestimation of fecal and UNE in the diets
with greater protein content, as calculations may not take into account fecal metabolic
nitrogen or urine endogenous nitrogen.
The percentage of retained nitrogen relative to the NI increased linearly ( P o 0.05)
with decreasing levels of dietary protein, pointing to increased efficiency in the utilization
of nitrogen. The abovementioned parameter corresponds to the NPU; an NPU value of
100% means that all dietary nitrogen was deposited into body tissues, and a value of 0%
means that none of the supplied nitrogen was transformed into protein. The values
encountered in this study ranged from 70.83% to 78.44%, indicating that the dietary
nitrogen was better used by the piglets with the lowest protein level. Kerr and Easter
(1995) found NPU values of 57.50% and 68.28% in animals fed diets without and with
amino acids, respectively (12% of CP).
The biological value of the food protein (BVFP) corresponds to the percentage of
apparently absorbed nitrogen retained by the organism to build or repair tissues. Proteins
with high biological value or complete proteins contain all of the EAA in the amounts and
proportions necessary for meeting the organic requirements of the animal. The proteins
with greater EAA contents exhibit better digestibility and promote more efficient intestinal
absorption. The biological value of the dietary protein in the present study varied from
91.25% for the diet with the lowest CP content (15%) to 79.87% for the diet with the
highest CP content (21%), thus showing that the availability of amino acids in all diets
was appropriate.
The reduction of nitrogen in the diets was correlated with lower levels of amino acid
catabolism, resulting in reduced levels of urea. This response was expected and is in
agreement with the findings of other authors ( Gómez et al., 2002 ; Kerr et al., 2003 ).
Urea is generally produced when protein-rich diets are fed and when the amino acid
intake surpasses the amount needed by the organism for the synthesis of proteins; urea
is also produced under fasting conditions when the cellular proteins are used as energy
sources ( Nelson and Cox, 2011 ). Urea levels also increase when certain amino acids are
lacking in the diet, thus altering the speed of protein
synthesis ( Penz Jr. and Viola, 1998 ). Therefore, plasma urea levels increase due to either
an amino acid unbalance or an intake of low-quality protein ( Coma et al., 1995 ). In the
present study, the highest plasma urea levels were found in animals fed diets consisting
of 21.0% and
19.5% of CP, possibly due to an imbalance in the amino acid absorption profile.
Albumin levels alter in response to variations in protein intake only after a relatively
long period of time. The relatively short time frame of the experiments may therefore
explain the lack of significant differences among the treatments.
According to Figueiredo et al. (2003) , when the globular
proteins (total protein, albumin, and globulin) decrease, the availability of amino acids for
protein synthesis is also reduced. As related parameters did not exhibit statistically
significant differences among treatments in the present study, it is likely that the use of
synthetic amino acids was efficient in promoting appropriate synthesis of proteins.
Unlike carbohydrates and lipids, excess amino acids cannot be stored in the tissues.
These amino acids are catabolized and transformed into toxic ammonia, which is
subsequently transformed in glutamine and circulated to the liver; in the liver, glutamine is
transformed into urea and transported by the bloodstream to the kidneys, where it is
filtered and excreted with the urine ( Wright, 1995 ). Urea represents 95% of the total urine
nitrogen ( Canh et al., 1998 ).
In the present study, the diets with higher CP contents (21.0% and 19.5%) promoted
higher urine nitrogen levels, thus corroborating the observation that excess protein results
in the superfluous accumulation of the amino acids tryptophan, isoleucine, and valine,
which are consequently excreted in greater volumes. Thus, an amino acid imbalance
occurred, with valine and isoleucine surpassing the physiological requirements of the
animals. The diet consisting of 15% CP exhibited the ideal amino acid profile. These
results confirm that the measurement of urea concentrations is a useful auxiliary method
for the establishment of the protein requirements of swine, as proposed by Chen et al.
(1995) .
The increased creatinine concentrations found in the present study were due to a
reduction in the volume of excreted urine associated with reduced water intake by the
piglets fed diets with decreasing levels of CP. The urine creatinine concentrations
increased with decreasing urine volumes.
According to Kaneko et al. (1997) , as a product of
nitrogen metabolism that cannot be reabsorbed or reutilized by the organism, creatinine is
continuously removed via the kidneys. Urine creatinine levels are highly variable and may
be influenced by dietary factors, whereas the daily creatinine excretion is proportional to
the weight of the pigs ( Deguchi, 1998 ).
5. Conclusions
The reduction of the CP content from 21% to 15% in diets supplemented with
synthetic amino acids did not affect the performance of piglets weighing 6 – 15 kg. The
EAA:NEAA ratio increased with decreasing levels of dietary
 J.B. Toledo et al. / Livestock Science 168 (2014) 94 – 101
CP, and the best ratio (53:47) was found in the diet containing 15% CP. The blood urea
concentrations and the UNE decreased with dietary protein levels and thus appeared to
be correlated with the CP content of diets. From an environmental perspective, reduction
of the dietary CP is an efficient method of reducing the polluting effect of pig slurry.
Acknowledgments
The authors acknowledge the National Council of Scientific and Technological
Development (Conselho Nacional de Desenvolvimento Científico e Tecnológico –
CNPq) for the fellowship award and the Ajinomoto Biolatina company for supplying the
ingredients and performing the necessary tests for the present study.
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