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The study was conducted in Field 2. Two varieties Napier grass used in this experiment. The varieties were Common Taiwan Napier (Pennisetum purpureum) and unknown variety of Taiwan Napier from Sabah. They were planted using stem cutting.
3 .2 Experimental Designs
The study area was about 180 m² which was divided into 24 plots. The size of a plot was 7.5 m² (3 m x 2.5 m). Planting distance of every plot is 50 cm (within rows) x 60 cm (between rows). Experimental unit group called block or replication using. The material was planted in a Randomized Complete Block Design (RCBD), with 4 replications, 3 treatments, and 2 varieties. The layout experimental unit group is shown in figure 1.
N1 = 0 kg N/ha (control)
N2 = 100 kg N/ha
N3 = 200 kg N/ha
V1 = dwarf Napier
V2 = common Napier
R1 = replication 1
R2 = replication 2
R3 = replication 3
R4 = replication 4
*The number of plot that is going to be used: 4 x 3 x 2= 24 plots
Randomized complete block design:
Factorial experiment 3 treatments x 2 varieties x 4 replications
Figure 1: layout of experimental unit group design
3.3 Fertilization Treatment
Three treatment consist of nitrogen fertilizer in form of urea (N) was applied to plots at rates of 0,100, and 200 kg haË‰¹ after each harvesting period. Urea also applies at the beginning of planting, together with phosphorus fertilizer (P) in form of Triple Super Phosphate (TSP) and potassium of fertilizer (K) in form of Muriate of Potash (MOP), at a rate of 50 kg haË‰¹ during land preparation for a basal fertilizer.
3.4 Harvest and Collection of Forages Samples
In order to standardize, all plots were cut at 70 days after planting without data collection. The grasses were left to grow for 4weeks before it can harvest. To minimize the effect of border row, all samples was taken 0.5 square meter (1 x 0.5 m quadrate) area as sample for each plot. There should ideally were 3 clumps within that area because planting distance within a row was 50 cm and between row was 60cm. The grasses were cut 30 cm from the ground and each sample weight was taken and put on paper bags and label the bags as follows:
R1 V1 N1 H1
R1 = Replication 1
N1 = fertilization (treatment)
H1 = date of harvest
3.5 Yield Analysis
All the grasses samples were weight each 500 g to obtain the fresh matter weight and were cuts into small pieces with a pair of scissors. Then, all the samples were dry in the oven for 3 days at 65Ëšc. After that, all weight of samples was taken to determine the dry matter on offer, which determines the yield at one time. The averages of dry weight sample were converted to kg/ha.
3.6 Chemical Analysis (Measure Nutritive Quality on Plant)
3.6.1 Samples Grinding
For doing the analysis, all the samples must to be grinding at the suitable particle size. The samples were put in grinder machine to obtain small particle size. Then, the processes were repeated using small grinder machine to obtain the right particle size for chemical analysis. Lastly, all the samples were put in the bottle for storage and it were labeled.
3.6.2 Crude Protein Analysis by Kjeldahl Method
The proximate analysis was used to determine measure the nitrogen (N) content of the grass samples. It is referred as crude because the measurement of protein is derived from the estimation of the nitrogen (N) content of the grasses. It means that technique does not differentiate the sources of N whether it derives from true or non-protein compounds found in the grasses. All protein contains N and not all that is N is a protein. The N content measured by the technique is then multiplied with a common factor, 6.25 to obtain the percentage of crude protein of samples. The methods were done from 0.5-1.0 g samples. Each sample was places and pour into the kjedahl flask and add ½ - 1 catalyst tablet was put and pour carefully 10 ml concentrated H2SO4 acid and 3-5 ml H2O2 solution into the flask. Digest the samples until the content becomes clear or straw colour. After that, digested the solution into 250 ml volumetric flask and add 250 ml with distilled water.
3.6.3 Fiber and Lignin Analysis by Van Soest Method
(C-b) / a *100
The Van Soest detergent system was used to determine the fiber concentration of samples. The method procedures separates the grasses into two component namely cell walls and cell contents. Neutral detergent fibre (NDF) and acid detergent fibre (ADF) were done from 0.5-1 g samples and put into a digestion tube. Before that, weight of empty digestion tube were taken and recorded. After that, all the digestion tubes were arranges into hot plate and pour with solution until the samples was covered. The difference NDF and ADF was solutions were used. Then, 2 or 3 drops of decahydronaphthalene was put into solution and the samples were heated for 1 hour. Meanwhile, distilled water was heated until it reaches 80Ëšc. After finish the heated processes, all the samples were move out from hot plate and washed it twice with hot water and once with acetone. Then, all the samples were dried in the oven at 130Ëšc for 2 hour. After that, samples were weighed to obtain the weight of dried digestion tube with sample. Percentage of NDF and ADF content are calculated using the formula below:
a: weight of feed sample
b: weight of empty capsule
c: weight of dried capsule with sample residue
(C-b) / a *100
Finally, following the procedure for the determination of ADF, the estimation of acid detergent lignin (ADL) is carried out. The acid detergent solution solublises the protein and all other soluble compound in feed sample. Residues of ADF consist of lignocelluloses and acid insoluble ash. Strong acid will dissolve the cellulose component and ashing of this residue will determine the lignin component of the feed. ADL content was calculated using the formula below:
a: weight of feed sample
b: weight of dried capsule with sample residue
c: weight of dried capsule with ash
3.7 Data Analysis
The experiment was conducted using the Randomized Complete Block Design (RCBD). Data collected were subjected to Analysis of Variance (ANNOVA) followed by LSD to determine significant differences (P<0.05) among the mean. All the data was analysis using a general linear model (GLM) procedure of the Statistical Analysis System (SAS). The GLM procedure can perform simple or complicated ANOVA for balanced or unbalanced data. Four types of estimable functions of parameters are available for testing hypotheses in PROC GLM. For data with no missing cells, the Type III and Type IV estimable functions are the same and test the same hypotheses that would be tested if the data were balanced. Type I and Type III sums of squares are typically not equal when the data are unbalanced; Type III sums of squares are preferred in testing effects in unbalanced cases because they test a function of the underlying parameters that is independent of the number of observations per treatment combination.