International Journal of Applied and

Natural Sciences (IJANS)

ISSN (P): 2319–4014; ISSN (E): 2319–4022
Vol. 10, Issue 1, Jan–Jun 2021; 71–80
© IASET

EFFECTS OF PLANT SPACING AND FERTILIZER LEVEL ON CHEMICAL

COMPOSITION OF HYBRID BRACHIARIA CV. MULATO II GRASS DURING THE FIRST
150 DAYS OF GROWTH UNDER IRRIGATION SUPPLEMENTATION, IN CHAGNI
RANCH, AWI ZONE, ETHIOPIA

Wondimagegn Tadesse1, Berhanu Alemu2 & Mesganaw Addis3

1
Research Scholar, Department of Animal Production and Technology, College of Dry Land Agriculture, Kebri Dehar
University, Kebri Dehar, Ethiopia
2,3
Research Scholar, Department of Animal Science, College of Agriculture and Natural Resources, Debre Markos
University, Debre Markos, Ethiopia

ABSTRACT

A study was conducted to evaluate the effects of plant spacing and N fertilizer application on chemical composition of
Brachiaria hybrid cv. Mulato II grass for the first 150 days after planting. A factorial experiment with 3 urea fertilizer
levels (0, 50 and 100 kg/ha) and 4 spacings between plants and rows (20 x 20, 30 x 40, 40 x 60 and 50 x 80 cm) with 3
replications was used. Chemical analyses were conducted for crude protein (CP), ash, neutral detergent fiber (NDF), acid
detergent fiber (ADF) and acid detergent lignin (ADL). Results indicated that DMY, CP%, CPY, NDF% and ADF% were
significantly (P<0.05) affected by plant spacing and fertilizer levels interactions. However, ash and ADL were significant
(P<0.05) affected by only main effects. The highest CP% was recorded for wider plant spacing (50 x 80 cm) with higher
urea fertilizer level (100 kg/ha) (S4F3) Similar studies need to be conducted over much longer periods to determine to
what extent these findings relate to performance over the life of a permanent pasture.

KEYWORDS: Urea; Spacing; Dry Matter Yield; Chemical Composition

Article History
Received: 23 Apr 2021 | Revised: 28 Apr 2021 | Accepted: 30 Apr 2021

INTRODUCTION

Livestock are an important component of nearly all farming systems in Ethiopia, providing milk, meat, draught power,
transport, manure, hides and skins and serve as a source of cash income (Funk et al. 2012). The subsector contributes about
16.5% of the national Gross Domestic Product (GDP) and 35.6% of the Agricultural GDP. It also contributes 15% of
export earnings and 30% of agricultural employment. The livestock subsector currently supports and sustains livelihoods
for 80% of the total rural population (Samson and Frehiwot 2014). Despite the importance of livestock in the country,
productivity is low (Sintayehu et al. 2010). One of the major constraints leading to such low productivity is shortage of
feed in terms of both quantity and quality, especially during the dry season (Ahmed et al. 2010) combined with high feed
prices (Sintayehu et al. 2010).

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72 Wondimagegn Tadesse, Berhanu Alemu & Mesganaw Addis

In order to solve the shortage of feed and increase livestock productivity, it is necessary to introduce and cultivate
high-quality forages with high yielding ability and adaptation to the biotic and abiotic environmental stresses (Kahindi et
al. 2007). Improved grasses, many of African origin, have greater palatability and productivity than other indigenous
species and are therefore desirable additions to pastures and common grazing areas (Alemayehu 2002). Among the
improved forage crops introduced into Ethiopia, Mulato II grass, which is the result of crosses of Brachiaria ruziziensis, B.
brizantha and B. decumbens, is claimed to play an important role in providing a significant amount of quality forage (CIAT
2006).

The optimization of production and nutritive value of grass can be achieved by planting on fertile soils (ILRI
2010) and forage management tools such as plant spacing (Sumran et al. 2009). Nitrogen fertilizer application is a common
practice since this nutrient is found to be one of the most limiting factors influencing yield and chemical composition of
grass pasture including crude protein (CP) concentration and digestibility, which in turn improves livestock production
(Marques et al. 2017). Nevertheless, information regarding the effects of fertilizer levels and plant spacing chemical
composition of Mulato II grass is scarce in our country and specifically in the study area. We conducted the present study
in order to generate information on chemical composition of Mulato II grass at different spacings with different rates of
nitrogen fertilizer.

MATERIALS AND METHODS

Description of the Study Area

The experiment was conducted in Chagni Ranch, Guangua Woreda, Awi Zone, Amhara National Regional State, Ethiopia.
Chagni (10°57′N, 36°30′E; elevation 1,583 masl), located at 528 km from Addis Ababa and 186 km West of Regional
town, Bahir Dar, is the administrative center of Guangua District (Asnake 2009). The Ranch is located about 500 m south
of the town. The area has average annual rainfall of 1,689 mm and mean minimum and maximum annual temperatures of
23 oC and 30 oC, respectively (Chagni ranch office).

Experimental Layout, Design and Treatments

The study was conducted using a 3 x 4 factorial arrangement in a randomized complete block design (RCBD) with 3
replications. The factors were 3 levels of urea fertilizer (0, 50 and 100 kg/ha) and 4 spacings (20 x 20, 30 x 40, 40 x 60 and
50 x 80 cm) between plants and rows, respectively, giving 12 treatment combinations (Table 1) and 36 experimental plots.

Table 1: Treatment Combinations

Spacing
Fertilizer Level
S1 S2 S3 S4
F1 F1 X S1 [T1] F1 X S2 [T2] F1 X S3 [T3] F1 X S4 [T4]
F2 F1 X S1 [T5] F2 X S2 [T6] F2 X S3 [T7] F2 X S4 [T8]
F3 F3 X S1 [T9] F3 X S2 [T10] F3 X S3 [T11] F3 X S4 [T12]
S1 = 20 x 20 cm spacing; S2 = 30 x 40 cm spacing; S3 = 40 x 60 cm spacing; S4 = 50 x 80 cm spacing between plants
and rows, respectively; T = treatments 1-12; F1= 0 kg urea/ha; F2 = 50 kg urea/ha; F3 = 100 kg urea/ha.

The size of each plot was 3 m long by 3.2 m wide with a gross plot size of 9.6 m2 and the total experimental area
was 12.6 m by 41.5 m (522.9 m2). The spacings between plots and replications were 0.5 and 1.5 m, respectively.
Treatments were randomly assigned to plots within each replication.

Impact Factor (JCC): 6.3238 NAAS Rating 3.73

Effects of Plant Spacing and Fertilizer Level on Chemical Composition of Hybrid Brachiaria CV. Mulato II Grass during 73
the First 150 Days of Growth Under Irrigation Supplementation, in Chagni Ranch, Awi Zone, Ethiopia

Land Preparation, Experimental Management, Soil Sampling and Analysis

Land was oxen-ploughed, and harrowing and bed preparation were carried out before planting manually. Root splits of
Mulato II grass were collected from Finota Selam grass nursery site at an age of 7 months re growth and planted at the
experimental site on 6 September 2017. Urea was purchased from the local market and applied by split application at time
of planting and again 30 days after planting with different levels based on treatment. The 25kg/ha for treatment taking
50kg/ha and 50kg/ha for treatment taking 100kg/ha of urea was applied at time of plainting and again 30 days if planting.
Weeding was done manually during the experimental period. The experiment was irrigated once a week when rain was
limited, with precautions taken to avoid contamination of treatments by cross flooding. Soil samples were taken by auger
from the center and corners of the experimental site prior to planting and from the individual plots immediately after
harvesting to a depth of 15 cm. The collected samples were thoroughly mixed, dried, ground and preserved in plastic bags
for chemical analysis to evaluate total nitrogen, available phosphorus, pH, organic matter and organic carbon. Total N was
determined using the Kjeldahl procedure (Bermner and Mulvaney 1982) and available P using the Olsen method (Olsen et
al. 1954). The totan organic carbon of soil was determined based on the Walkely-Black chromic acid wet oxidation
method. Organic matter (OM) was calculated indirectly from organic carbon (OC) concentration by multiplying OC by
1.724 and the pH was determined using the method described by Van Reeuwijk (1993).

Chemical Analysis of Forage

The forage samples were thoroughly mixed and a 0.5 kg fresh subsample was taken from each plot for chemical analysis
and dried in a forced-draft oven at a temperature of 105 ºC for 24 hours. The dried material was ground to pass through a 1
mm sieve for chemical analysis and preserved in a plastic bag pending chemical analysis at Debre Birhan Agricultural
Research Center Animal Nutrition Laboratory. Ash and nitrogen (N) were determined according to the procedures
described by AOAC (1990). Neutral detergent fiber (NDF), acid detergent fiber (ADF) and acid detergent lignin (ADL)
were determined according to the procedures described by Van Soest and Robertson (1985). .

Statistical Analysis

Data were subjected to analysis of variance (ANOVA) using the General Linear Model (GLM) procedure of the Statistical
Analysis System (SAS 2007). Differences among treatment means were determined using Duncan’s Multiple Range Test
(DMRT), at P<0.05. The statistical model used was:

Yijk = µ + Bi + Fj + Sk + (FS)jk + eijk ,

Where:

Yijk = the response variable

µ = over all mean

Bi = ith block effect

Fj = jth main factor effect (Fertilizer level)

Sk = kth main factor effect (Spacing)

(FS)jk = jkth interaction effect (Fertilizer Level x Spacing)

eijk = random error.

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74 Wondimagegn Tadesse, Berhanu Alemu & Mesganaw Addis

RESULTS

Soil Analysis Report

Soil samples taken before planting of grass revealed the following: organic matter (OM) – 5.88%; organic carbon (OC) –
3.41%; total N – 0.30%; available P – 4 ppm; and pH – 5.6. After harvesting total N of the soil increased slightly with the
highest level of urea as compared with the value before planting (Table 2), while pH declined. Available phosphorus (Av.
P) declined on all treatments, while both OC and OM concentrations increased (Table 2).

Table 2: Soil Analysis Report After Forage Harvest for Various Urea Applications
Soil Parameters
Treatment pH Av. P (ppm) OC (%) OM (%) Total N (%)
0 kg urea/ha 5.68 2 3.39 5.84 0.29
50 kg urea/ha 5.53 2 3.48 6.00 0.30
100 kg urea/ha 5.48 2 3.85 6.64 0.33
SSBP 5.6 4 3.41 5.88 0.30
SSBF = Soil Sample Before Planting; Av. P = Available Phosphorus; OC = Organic Carbon; and OM =
Organic Matter.

Chemical Composition Brachiaria Hybrid (Mulato II) Grass

The significant effects of N fertilizer levels, spacing and their interactions on crude protein percentage (CP) and crude
protein yield (CPY) is indicated in Table 3. CP concentration increased (P<0.05) with increase in row spacing at all
fertilizer levels and urea application increased C at all plant spacings but differences were significant (P<0.05) at only the
narrowest and widest spacings. Similarly, urea application increased CPY at all plant spacings but differences were
significant (P<0.05) at only the narrowest spacing. Highest CP% (20.0%) was recorded where 100 kg urea/ha was applied
at the widest plant spacing and the lowest (14.6%) for the Control treatment at the narrowest plant spacing. In contrast,
highest CPY (1.45-1.57 t/ha) was recorded where urea was applied at the narrowest plant spacing.

Table 3: CP Concentration and Crude Protein Yield (CPY, t/ha) of Mulato II Grass as Affected by Urea
Fertilizer Level, Plant Spacing and their Interactions
CP (%) CPY
Spacing Fertilizer Level Fertilizer Level
F1 F2 F3 F1 F2 F3
S1 14.6h 16.2g 17.2defg 0.95cd 1.45ab 1.57a
S2 16.6fg 16.6fg 18.1bcdef 0.86cd 0.94cd 1.17bc
S3 17.7cdefg 18.3bcde 19.1abc 0.74de 0.83de 0.87cd
S4 18.5bcd 19.5ab 20.0a 0.53e 0.78de 0.74de
S1 = 20 x 20 cm; S2 = 30 x 40 cm; S3 = 40 x 60 cm; and S4 = 50 x 80 cm spacing between plants and rows; F1= 0
kg urea/ha; F2 = 50 kg urea/ha; and F3 = 100 kg urea/ha.

Means for different treatments with different letters are significantly different (P<0.05).

Neutral Detergent Fiber (NDF) and Acid Detergent Fiber (ADF)

The significant effects of N fertilizer levels, spacing and their interactions on neutral detergent fiber (NDF) and acid detergent
fiber (ADF) are presented in Table 4. Both NDF and ADF concentrations declined as fertilizer level increased but differences
were significant (P<0.05) only for the narrower two plant spacings. Similarly, NDF% declined as plant spacing increased but
differences were significant (P<0.05) only in the Control treatment. ADF% also declined as plant spacing increased but there
were no consistent significant differences. Overall trends were for highest values for both NDF and ADF concentrations to occur
in the Control at the narrowest plant spacing and the lowest values in the higher urea level at the widest plant spacing.

Impact Factor (JCC): 6.3238 NAAS Rating 3.73

Effects of Plant Spacing and Fertilizer Level on Chemical Composition of Hybrid Brachiaria CV. Mulato II Grass during 75
the First 150 Days of Growth Under Irrigation Supplementation, in Chagni Ranch, Awi Zone, Ethiopia

Table 4: NDF and ADF Concentrations of Mulato II Grass as Affected by Urea Fertilizer Level, Plant
Spacing and their Interactions
NDF (%) ADF (%)
Plant Spacing Fertilizer Level Fertilizer Level
F1 F2 F3 F1 F2 F3
S1 52.8a 48.5abc 45.1cde 39.3a 36.9abc 33.7bcde
S2 51.0ab 47.8bcd 42.1e 38.2ab 36.3abcd 32.1cde
S3 48.3abc 43.6cde 46.6bcde 36.3bcd 30.8de 31.0de
S4 45.5bcde 46.2bcde 42.3de 34.4bcde 33.3bcde 30.1e
S1 = 20 x 20 cm; S2 = 30 x 40 cm; S3 = 40 x 60 cm; and S4 = 50 x 80 cm spacing between plants and rows; F1= 0
kg urea/ha; F2 = 50 kg urea/ha; and F3 = 100 kg urea/ha.

Means for different treatments with different letters are significantly different (P<0.05).

Ash and ADL content

As there was no significant interaction between fertilizer level and plant spacing for ash and ADL concentrations, main
effect means are presented in Table 5. Ash concentration increased as fertilizer level increased (P<0.05) and as plant
spacing increased (P<0.05), while the reverse was the case for ADL with ADL% decreased as fertilizer level increased and
plant spacing increased (P<0.05).

Table5: Ash and ADL Concentrations of Mulato II Grass as Affected by Urea Fertilizer Level and Plant
Spacing
Fertilizer Level Plant Spacing
Parameter F1 F2 F3 Mean S1 S2 S3 S4 Mean
Ash (%) 9.86b 10.4ab 10.95a 10.4 9.43c 10.07bc 10.88ab 11.23a 10.4
ADL (%) 9.36a 8.56b 7.82b 8.58 9.55a 8.65ab 8.39ab 7.74b 8.58
S1 = 20 x 20 cm; S2 = 30 x 40 cm; S3 = 40 x 60 cm; and S4 = 50 x 80 cm spacing between plants and rows; F1= 0
kg urea/ha; F2 = 50 kg urea/ha; and F3 = 100 kg urea/ha.

Means for different treatments with different letters are significantly different (P<0.05).

DISCUSSION

Chemical Composition of Brachiaria Hybrid (Mulato II) Grass

Mulato II showed good response to fertilization, particularly to the application of nitrogen. Depending on the level of soil
fertility, one or more maintenance applications may be required to maintain high yields of good quality forage. The
percentage of croude protein (CP), acid detergent fiber (ADF), neutral detergent fiber (NDF), invitro dry matter
digestibility (IVDMD), depends on the soil fertility. Mulato II grass planted at wider spacing with higher N fertilizer level
had excellent nutritional value, particularly CP concentration, limiting nutrient in forages. Even forage planted at all
spacing with urea fertilizer application had CP concentration well above, the level of CP required for rumen function 7.5%
(Van Soest 1982) and for lactation 15% (Norton 1982). This is a clear indication of the value of this particular grass to be
used as valuable forage for livestock. The reason for the high CP percentage at higher fertilizer and wider spacing might be
due to emergence of younger plants, enhancement of leafiness and leaf to steam ratio of grass which has high protein
content than steam due nutrient and space availability. This result agree with finding Marques et al. 2017 who reported as
Nitrogen is essential among nutrient to gate significant increase in forage and to have a linear increase in CP rate of the
Mulato II grass

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76 Wondimagegn Tadesse, Berhanu Alemu & Mesganaw Addis

The crude protein yield (CPY) was increased with increasing of N fertilizer levels and reducing of spacing.
Generally, the CPY was found to be higher for application of nitrogen fertilizer at narrower spacing (20x20 cm). The
increment of CPY due to urea, in the current experiment was supported by Paulo et al. (2014) who recorded higher CPY
from Rye grass fertilized by higher level of N. Despite the reduction of CP percentage with narrower spacing, CPY
increased significantly as spacing become narrow. This might be due to, at closer space, DMY was higher with high dense
plant population since the CPY is a product of CP percentage and DMY.

Neutral detergent fiber (NDF) and acid detergent fiber (ADF) concentrations all declined as N fertilizer level and
spacing increased. While increase steam percentage, maturity and increased lignifications would account for the narrower
spacing without N fertilizer application. At lower fertilizer levels and narrower spacing’s resource limitations is high which
constrained the plants and make them to have higher structural components to cope with the stress full environmental
condition. The narrower spacing’s makes the plants to have higher structural materials and have higher NDF compared to
the wider spacing which had less resources limitations and had higher leaf numbers and leaf to steam ratio which are
assumed to be the factor for lower NDF and ADF content.

The current result is in line with Marques et al. (2017), who reported as Higher fertilizer dose led to higher
protein, lower NDF and ADF levels of Mulato II grass, which could increase nutritive of value grasses and its intake ().
Castagnara et al. (2011b) also detected the influence of N doses studied for NDF rates on three tropical grasses and four N
doses (0, 40, 80, 160 kg/ha) woth ureas as sources, and registered that the grasses had a quadratic behavior for the variable
NDF. The lowest rate was obtained with higher N doses. Conversely, Hazary et al. (2015) on Jumbo grass did not find any
significant effect on the NDF and ADF content with increasing level of N fertilizer. Berihun (2005) on Bana grass reported
similar NDF and ADF increment trend with the current study at narrow spacings. However, Genet et al. (2017) on Desho
grass reported higher NDF and ADL contents at wider spacings with the reasons being development of larger tillers in the
wider-spaced plants.

The increase in cell wall contents (NDF) is a very important limiting factor of nutritive value of feeds (Van Soest
and Robertson 1985). The NDF value greater than threshold level, 60% resulted in decreased voluntary feed intake,
increased rumination time and decreased conversion efficiency of ME (Reed and Goe 1989). Roughage diets with NDF
content of 45-65% and below 45% were generally considered as medium and high quality feeds, respectively (Singh and
Oosting 1992). Additionally, roughages with less than 40% ADF content are categorized as high quality and those with
greater than 40% as poor quality (Kellems and Church 1998). Mulato II grass established at different spacing and fertilizer
level in the current study were bellow the critical value 60% NDF that decreased voluntary feed intake, feed conversion
efficiency, longer rumination time (Reed and Goe 1989) and 40% ADF which considered as high quality grasses (Kellems
and Church 1998).

The ash content in any feed is a positive indicator of the inorganic (minerals) content was significantly higher at
higher N fertilizer level (F3). It was increased as the fertilizer levels increases, might be due to higher urea levels facilitate
green leave production which uses more mineral for different physiological purposes and nitrogen fertilizer increased the
mineral content in leaves. The observed high content of ash at higher levels of fertilizer was supported by the findings of
Joorabi et al. (2014) who obtained higher mineral contents in Sorghum crops grown on higher fertilizer levels. Contrary to
the current study, Abdi (2014) on Cenchrus ciliaris and Panicum maximum and Hazary et al. (2015) on Jumbo grass
reported lower ash contents grasses with higher doses of nitrogen fertilizer levels. Total ash content was being reduced as

Impact Factor (JCC): 6.3238 NAAS Rating 3.73

Effects of Plant Spacing and Fertilizer Level on Chemical Composition of Hybrid Brachiaria CV. Mulato II Grass during 77
the First 150 Days of Growth Under Irrigation Supplementation, in Chagni Ranch, Awi Zone, Ethiopia

spacing becom narrow. This might be due to less competition among plants for minerals resources uptake from soil and
that facilitate emergence of more leaves. The current result support the report of Genet et al. (2017) where increase in ash
contents for Desho grasses grown on wider spacings.

The application of N fertilizer decreased the ADL content of the grasses, might be due to the continuous
development of younger plants at N fertilizer application which could increase leafiness of grasses that in turn increases
leaf to steam ratio. The urea fertilizer promotes the development of new leaves and shoots with low lignin content. When
lignin is lowered digestibility plants increased since lignin is highly resistant to chemical and enzymatic degradation by the
micro flora of the rumen. According to Abdi (2014), urea promotes the growth of new leaves and shoots resulting in low
ADL contents of Cenchrus ciliaris and Panicum maximum grasses. The ADL content decreased as spacing increased,
might be due to emergence of younger tillers from wider spacings which makes the plants to bear more leaves as a result of
less competition for soil nutrient at wider spacing. Similarlly, Berihun (2005) on Bana grass reported higher plant density
per unit area increased the value of ADL.

The current result revealed that, quality of Mulato II grass can be improved by urea fertilizer application. More
specifically the application of 100 kg/ha urea fertilizer is the most important to provides higher CP% and lower fiber
content with better economic feasiblity. On the other hand, CP% of grasses at wider spacings 50x80 cm were found to have
high values. An important limitations of this study, that could be addressed in future works are:

• The trial needs to be conducted across a years to reach a complete recommend of optimum chemical composition
more precisely.

• Further study on re-growth yield of Mulato II grass is useful to determine continuing productivity of grass in
terms of chemical composition.

• The trial needs to be conducted for different types of soil to recommend optimum chemical composition of
Mulato II grass.

• Further study on harvesting time and fertilizer type effects on Mulato II grass is useful to determine productivity
of the grass in terms of chemical composition.

• The magnitude of improvement in the chemical composition of Mulato II grass needs to be studied in terms of
animal performance.

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