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Cuban Journal of Agricultural Science

Print version ISSN 0864-0408On-line version ISSN 2079-3480

Cuban J. Agric. Sci. vol.50 no.2 Mayabeque Apr.-June 2016


Cuban Journal of Agricultural Science, 50(2): 259-266, 2016, ISSN: 2079-3480




Effect of different nitrogen levels and irrigation techniques on the ruminal degradation of the crude protein of maize


Efecto de diferentes niveles de nitrógeno y técnicas de riego en la degradación ruminal de la proteína bruta del maíz



Barajas R.,I Díaz T.,II Flores L.R.,II Partida L.,II Martínez M.,III Lomelí J. J.,I Velázquez T de J.,II

IFacultad de Medicina Veterinaria y Zootecnia de la Universidad Autónoma de Sinaloa, Culiacán, Sinaloa, México.
IIFacultad de Agronomía de la Universidad Autónoma de Sinaloa. Apdo. Postal 1057, Culiacán, Sinaloa, México, CP 80000 .
IIIInstituto de Ciencia Animal. Apartado Postal 24. San José de las Lajas, Mayabeque, Cuba.




In order to determine the effect of ruminal degradation of maize crude protein obtained with two levels of nitrogen (300 and 400 kg ha-1) and two irrigation techniques (gravity and drip), dacron bags (10 x 18 cm) which contained 5 g of maize were used. By introducing a cannula, the bags were inoculated for 0, 3, 6, 12 and 24 h in the rumen of four heifers. A completely randomized design was applied with factorial arrangement 2x2 of the treatments (nitrogen and irrigation technique).There were not statistical differences between the interactions of the variables under study (P < 0.05).However, drip irrigation vs gravity (8.72 vs 8.05%) and fertilization with 400 vs 300 kg de N ha-1 (8.52 vs. 8.25 %) increased (P ≤ 0.05) the crude protein content of the grain. The treatments did not affect (P ≥ 0.21) the PC solubility (19.4 ± 2.85 %).The ruminal degradation of the maize CP in the different incubation times did not change (P ≥ 0.20) by the maize grain obtained by means of irrigation technique or N levels used in maize production. The ruminal degradation rate of CP (0.10 ± 0.03) did not change (P ≥ 0.13) by the treatments. The percentage of crude protein effectively degraded in the rumen (42.14 ± 2.83 %) was similar (P ≥ 0.48) between treatments. Although the schemes of nitrogen levels or irrigation technique can modify the crude protein content in the maize grain, this did not influence on the characteristics of ruminal degradation of its crude protein.

Key words: irrigation techniques, fermentation pattern, solubility.


Para determinar el efecto de la degradación ruminal de proteína bruta de maíz obtenido con dos niveles de nitrógeno (300 y 400 kg ha-1) y dos técnicas de riego (gravedad y goteo), se utilizaron bolsas de dacrón (10 x 18 cm) que contenían 5 g de maíz. Mediante la introducción de una cánula, las bolsas se inocularon durante 0, 3, 6, 12 y 24 h en el rumen de cuatro vaquillas. Se aplicó un diseño completamente al azar con arreglo factorial 2 x 2 de los tratamientos (nitrógeno y técnica de riego). No se presentaron diferencias estadísticas entre las interacciones de las  variables en estudio . Sin embargo, el riego por goteo vs gravedad (8.72 vs 8.05%) y la fertilización con 400 vs 300 kg de N ha-1 (8.52 vs. 8.25 %) incrementaron (P ≤ 0.05) el contenido de la proteína bruta del grano. Los tratamientos no afectaron (P ≥ 0.21) la solubilidad de la PB (19.4 ± 2.85 %). La degradación ruminal de la PB de maíz en los diferentes tiempos de incubación no se modificó (P ≥ 0.20) por el grano de maíz obtenido mediante la técnica de riego ni por los niveles de N usados en la producción de maíz. La tasa de degradación ruminal de PB (0.10 ± 0.03) no se alteró (P ≥ 0.13) por los tratamientos. El porcentaje de proteína bruta efectivamente degradada en rumen (42.14 ± 2.83 %) fue similar (P ≥ 0.48) entre los distintos tratamientos. Aunque los esquemas de niveles de nitrógeno o técnica de riego pueden modificar el contenido de proteína bruta en el grano de maíz, esto no influyó en  las características de degradación ruminal de su proteína bruta.

Palabras clave: técnicas de riego, patrón de fermentación, solubilidad.




The maize ranks third of cultivated area in the world, where more than 118 million hectares are planted with average production of about 600 million tons per year (Irfan et al. 2014) surface.

High doses of nitrogen fertilization increase the crude protein concentration in modern maize hybrids (Ferreira et al. 2005). Moreover, the uniform application and high irrigation doses modify the protein content in the grain, due to the ease of high production and assimilates translocation (Aydinsakir et al. 2013, Khalili et al. 2013).

Shah et al. (2003) found that drip irrigation produces a grain with high crude protein content than those found in maize irrigated by gravity. It is not known whether this change in the maize content has some influence on the ruminal degradation characteristics of maize protein. However, this grain is characterized by its importance as energy source for animal feeding, because it is rich in carbohydrates, as referred Ramalho et al. (2006). These authors showed that the starch sources are important in high producing animals, since they require high energy levels in the diet so that they can express their genetic potential.

The starch digestibility in dairy cows improves with maize diets, with mean particle size. However, although milk production and protein increases, the fat production in the milk is reduced (Danes et al. 2013). Protein supplements are an essential component of feed rations for cows of high dairy yield (Horký 2014). In addition, the modernization of agricultural practices, aimed to increase maize production, modifies, to some extent, the chemical composition of the grain, which leads to modify the diet for animal.

The objective of this experiment was to determine the nitrogen influence, at doses of 300 or 400 kg N ha-1, and gravity or drip irrigation on the characteristics of the ruminal degradation of crude protein of white maize.



The research was conducted at Facultad de Medicina Veterinaria y Zootecnia de la Universidad Autónoma de Sinaloa in Culiacan, Sinaloa, Mexico, located at 24° 50 'NL, 107 ° 26' WL and 57.34 m o.s.l high. The climate of this area is classified as dry tropical, with average annual temperature of 24.8 ° C, average annual rainfall of 645.6 mm and prevailing rains in summer. The annual average relative humidity is 68%.

A total of four Criollas x Simental heifers of 320 ± 20 kg average weight were used, with permanent ruminal cannula (C-4 Diamond®) of 10cm internal diameter. They were housed in individual pens (3.0 x 6.0 m) with feeder and drinker. Fifteen days before the experiment, the animals were adapted to intake a diet with 16.58 % of crude protein (CP); 7.10 MJ kg-1 of maintenance Net Energy (mNE), with relation 30:70 of forage: concentrate. The diet composition is show in table 1.

During the adaptation and development of measurements, the diet was offered ad libitum (105 % of the intake of the previous day) in a single ration (0800). The food was served fresh and those which not intake was daily took away. Heifers had permanent access to clean and fresh water. On the third day of the beginning of the adaptation phase, the animals were internally and externally wormed (Closantel®, Cheminova of Mexico and Clorfenvifos®, Laboratorios Novartis) also 5 mL of vitamins A, D and E (Vitaflow®; Virbac) was intramuscularly injected.

Samples of 20 kg of white maize (hybrid Pantera from Asgrow Company) were used, which came from an experiment developed in a semiarid climate with summer rains and mean annual precipitation of 800mm. The soil was clayey, with field capacity (FC) of 57.5%, permanent wilting point (PWP) of 36.7%, bulk density (Bd) of 1.27 g cm-3, organic matter content of 0.82% (up 60 cm depth) and total nitrogen content of 0.037%, with different types of management: nitrogen fertilization with 300 or 400 kg ha-1 and drip irrigation or gravity, which constituted the treatments of this experiment.

In order to evaluate the ruminal fermentation conditions during the test, soybean paste was included, when considering the available information about the performance of this ingredient in rumen. Samples were grounded at a particle size of 2 mm (Vanzant et al. 1998). In dacron bags (10 x 18 cm) 5 g of sample were placed on wet basis (13.21 mg cm-2 ratio). The bags were tied with a nylon string and a space of 10 cm between each pair was left. One of the bags, with a weight of 200 g, was placed at the end of the line. At the upper end of the line 70 cm to be tied to the top of the cannula were left. Samples (150 g) of each of the maize introduced into the bags and of the soybean paste were taken. The maize samples were dried in a forced air oven at 105 °C for 24 h (AOAC 2012) to determine the amount of dry matter introduced in the bags. The bags were incubated in the rumen for 3, 6, 12 and 24 h and placed in reverse order to the incubation time. It was taken as the start time the moment in which the animals received the food (0800). When incubation time finish, the bags were removed from the rumen and washed with water until they were clean (Nocek 1988).

Solubility was determined by immersing dacron bags in a 0.15 N solution of NaCl in distilled water at 39 °C for 5 min (Owens and Zinn 1982). The bags were dried in oven at 50 ºC for 48 h and weighed. The contents of a couple of bags was a composite sample, to which it was determined CP (N x 6.25 Kjeldahl) (AOAC 2012). With the results, the ruminal degradation of CP was calculated (Schneider and Flatt 1975).

The results of the ruminal degradation of crude protein (CP) to different incubation times were used to calculate the indicators of the kinetics of ruminal degradation with the negative exponential model proposed by Ørskov and McDonald (1979) by the equation P = a + b (1 - e-ct), where:

P – potentially degradable protein 

a – soluble fraction

b - insoluble fraction but degradable of CP in rumen, there was enough time for it

c – constant degradation rate of the fraction b

t – time

Once obtained the indicators, the CP effectively degraded in the rumen was calculated with the formula:

P = a + ((b c) / (c + k)) suggested by Ørskov and McDonald (1979), where "k" represents the rate of small particles flow through the rumen. According to the proportion of forage and concentrate of the diet offered to animals, k = 0.03 depending on the type of intake diet (Ørskov 1988).

Analysis of variance on the basis of a completely randomized design with factorial arrangement 2 x 2 was applied (Hicks 1973). A value of α = 0.05 to accept statistical difference was fitted. The calculations were performed using version 8 of the software package StatistixTM (Statistix 2003).



The results of variance analysis for the studied variables indicated that there was no interaction between the study factors (P > 0.05). However, the content of crude protein (CP) in the maize which received drip irrigation was higher (P ≤ 0.05) than the gravity irrigation (8.72 vs. 8.05%), which can be attributed to the efficiency of distribution of water and nutrients (N) with drip irrigation technique. These results agree with that informed by Shah et al. (2003). The maize that received the highest dose of N ha-1 (400 kg) had a higher content (P ≤ 0.05) of CP with respect to the fertilized with 300 kg ha-1 of N (8.52 vs. 8.25%). It is expected that at higher concentration of nitrogen in the soil, the amount translocated to the grain maize from the root system is higher, which increases the CP content. This result agrees with that found by Ferreira et al. (2005), who appreciated increase in protein content in the grain as increased the amount of N applied in four maize hybrids. The average value of 8.39% of CP found in this experiment in grains of white maize from the Pantera of Asgrow hybrid, although it was lower at 9.8 ± .55% attributed by the National Research Council (1996), they corresponds with the CP values between 8% and 9%, which are normally found in modern maize hybrids released from the nineties (Lykos and Varga 1995, Ferreira et al. 2005, Corona et al. 2006).

The fermentation pattern that showed the CP of the soybean paste in this experiment is described in the equation P = 31.78 + 53.38 (1 – e -.076 t); R2 = 0.96 (P ≤ 0.001). The solubility of 31.78% of the soybean paste CP was close to the values of 28.5% and 29% referred in others papers (Kirkpatrick and Kennelly 1987 Canbolat et al. 2007). The percentage degradation of CP in soybean paste at 24 h of incubation in rumen was 85.16%, 88.7% value close to that informed by Kirkpatrick and Kennelly (1987) in Holstein heifers. The fermentation pattern in rumen exhibited by soybean paste coincides with expectations according to the information available in the literature (Kirkpatrick and Kennelly 1987, Canbolat et al. 2007), so it is assumed that the conditions which prevailed in rumen during the test were normal. The results obtained with maize and the comparison between the different treatments can be considered reliable.

The influence of treatments on the ruminal degradation of maize CP is presented in Table 2. The different fertilization and irrigation schemes did not change (p ≤ 0.21) the solubility of the maize CP, which was as average 19.4 ± 2.85 %. This value was close to the 22.3% found by Lykos and Varga (1995) in ground yellow maize. The treatments had no effect (P ≤ 0.20) on the ruminal degradability of maize CP in any of incubation times. The average value (38.49 ± 4.18%) obtained at 12 h and the 49.71 ± 4.59 %, found at 24 h in this experiment, are close to 40 and 48 % estimated by Wanderley et al. (1993) in yellow maize at the same incubation times, after correcting the influence of microbial contamination. The degradation rate of the maize CP was similar (P ≤ 0.13) in the different treatments averaging 0.10 ± 0.03. This value is in the range between 0.08 and 0.10, established by the National Research Council (1996) for the cracked yellow maize. The maize CP effectively degraded in the rumen was similar (P ≤ 0.48) between the four treatments, with an average of 42.14 ± 2.83 %. This value, calculated on the basis of nylon bags technique, corresponds with 41.87 % determined by Prigge et al. (1978) in dry rolled maize, when measuring duodenal flow, and with 43% calculated by Stern et al. (1983) for maize gluten protein. Barajas and Zinn (1998), with measurements from the flow to the duodenum in heifers, estimated at 42.8% the ruminal degradation of the protein of dry cracked yellow maize. Corona et al. (2006) measured the flow to the duodenum in steers fed maize dry processing or steam- flaked from four hybrids, with CP content ranging between 8.1 and 8.7%. These authors found no influence of grain protein level on the ruminal degradation of CP, which estimated at 44.8%. The National Research Council (1996) attributed to dry rolled maize a content of 45% of CP degradable in rumen.

Although the fertilization and irrigation schemes allow modify the CP content of maize grain (Shah et al. 2003, Ferreira et al. 2005), apparently have no major effect on the rate or extent to which their proteins are degraded in the rumen , confirming the findings of Redd et al. (1975). These authors did not observed change in the amount of protein that came to abomasums in young bull fed with normal maize or opaque-2 maize, which is a variety with higher crude protein content. Harrelson et al. (2009) found no differences in ruminal degradation of six maize hybrids dry processed.

The difference in the crude protein content of the grain, as a result of the different management which was given to the maize, did not influence on the extent of protein degradation by ruminal microbiota.



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Received: 5/5/2015
Accepted: 11/7/2016



Barajas R., Facultad de Medicina Veterinaria y Zootecnia de la Universidad Autónoma de Sinaloa, Culiacán, Sinaloa, México. Email:

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