Effect of different study factors on the in situ

ruminal degradability of ammonified

sugarcane bagasse

Efecto de los diferentes factores de estudio en la

degradabilidad ruminal in situ del bagazo de caña

amonificado

Ricardo Daniel Vélez Saeteros

Ángel Ruisdael Bravo Vargas

Ernesto Antonio Hurtado

Edis Geovanny Macías Rodríguez

Abstract: This study evaluated the in situ degradation of ammonified

sugarcane bagasse silage as cattle feed during the dry season. Samples

of fresh sugar cane bagasse (Saccharum sp) were randomly taken

from the "Puro Bravo" farm, located in the "La Soledad" site, Junin

canton, province of Manabi. A Completely Randomized Design with

3x2 Factorial Arrangement was used, where Factor A was represented

by the different levels of urea inclusion (0, 3 and 5 %) and Factor B,

fermentation times of sugarcane bagasse (30 and 44 days), each

combination of these factors consisted of three replications. The best

treatment is 5% urea, day 30 of fermentation, both in bromatology

and degradability, and the degree of association of degradability at

different times was related to the nutritional value of the ammonified

bagasse, which is related to degradability and fiber. Ammonification

with urea improved nutritional values, characterized by increasing

crude protein and decreasing NDF, FDA and LDA as urea levels

increased. The differences were highly significant (P < .0001),

indicating that the results were highly significant for each of the

variables studied, which allowed relating the degree of association of

degradability at different times with the nutritional value of the

ammonified bagasse silage.

Keywords: Agribusiness, degradability, nutrition, dry season.

Published

Instituto Tecnológico Superior Edwards

Deming. Quito – Ecuador

Periodicity

April - June

Vol. 2, Num. 2, 2023

Dates of receipt

Received: January 09, 2023

Approved: March 01, 2023

http://centrosuragraria.com/index.php/revista

vol. 1. Num. 17. 2023.

pp. 13-25

Correspondence author

rvelez@uagraria.edu.ec

Creative Commons License

Creative Commons License, Attribution-

NonCommercial-ShareAlike 4.0

International.https://creativecommons.org/lice

nses/by-nc-sa/4.0/deed.es

1 Universidad Agraria del Ecuador, Ecuador - Guayaquil, rvelez@uagraria.edu.ec, https://orcid.org/0000-0001-

7720-9441

2 Universidad Agraria del Ecuador, Ecuador - Guayaquil, angel.ruisdael@gmail.com https://orcid.org/0000-

0003-4955-7099

3 Escuela Superior Politécnica de Manabí, Calceta, Manabí, ernesto.hurtado@espam.edu.ec

http://orcid.org/0000-0003-2574-1289

4 Universidad Técnica de Manabí, Ecuador - Portoviejo, edis.macias@utm.edu.ec, https://orcid.org/0000-

0002-1113-3031

Effect of different study factors on the in situ ruminal degradability of ammonified sugarcane

bagasse.

Resumen: Mediante este estudio se pudo evaluar la degradación in

situ del ensilaje del bagazo de caña de azúcar amonificado, como

alimento del ganado bovino en época seca. Se tomaron aleatoriamente

muestras de bagazo fresco de caña de azúcar (Saccharum sp) de la

Hacienda “Puro Bravo”, ubicada en el sitio “La Soledad”, cantón

Junín, provincia de Manabí. Se utilizó un Diseño Completamente

Aleatorizado con Arreglo Factorial 3x2, donde el Factor A se

representó por los diferentes niveles de inclusión de urea (0, 3 y 5 %)

y el Factor B, tiempos de fermentación del bagazo de caña (30 y 44

días), cada combinación de estos factores constó con tres repeticiones.

El mejor tratamiento es el de urea al 5%, día 30 de fermentación, tanto

en bromatología como en degradabilidad, además se relacionó el

grado de asociación de la degradabilidad en distintos tiempos con el

valor nutricional del bagazo de caña amonificado, el mismo que sí se

relaciona con la degradabilidad y con la fibra. La amonificación con

urea mejora los valores nutricionales, caracterizado por el

aumentando de la proteína cruda y diminución de, así como la FDN,

FDA y LDA en la medida de que se aumentan niveles de urea. Las

diferencias fueron altamente significativas (P <.0001) .Lo expuesto

indica que los resultados fueron altamente significativos para cada

una de las variables estudiadas, con lo cual se pudo relacionar el grado

de asociación de la degradabilidad en distintos tiempos con el valor

nutricional del ensilaje de bagazo de caña amonificado.

Palabras clave: Agroindustria, degradabilidad, nutrición, temporada

seca.

1. Introduction

Effect of urea level (0.3 and 5%) and different fermentation times (30

and 44 days) on the chemical composition of sugarcane bagasse. The

experiment was conducted at the Hacienda "Puro Bravo", located in the

site "La Soledad", canton Junín, province of Manabí; geographically

located between the coordinates 0º 56" 8" South latitude, 80º 11" 0"

West longitude and an altitude of 15 meters above sea level

(masl)(PDOT .2014).

The present experiment lasted three months, from the execution of the

project until the bromatological analyses were obtained.

FACTORS UNDER STUDY

FACTOR A: Urea levels (0, 3 and 5 %)

April - June vol. 2. Num. 2 - 2023

FACTOR B: Bagasse fermentation times (30 and 44 days)

EXPERIMENTAL DESIGN

A Completely Randomized Design with 3x2 Factorial Arrangement

was used, where Factor A was represented by the different levels of

urea inclusion (0, 3 and 5 %) and Factor B, cane bagasse fermentation

times (30 and 44 days). Each combination of these factors consisted of

three replicates.

The statistical model used was as follows:

yijk = µ + αi + βj + (αβ)ij + εijk

Where:

: k-th observation of the i-th level of factor A and j-th level of

factor B

General average.

Effect of the i-th level of factor A (inclusion of urea) i=1,2 and 3

Effect of the j-th level of factor B (Days of fermentation) j= 1 and 2

First-order interaction effect of the i-th level of factor A with the

j-th level of factor B.

Random effect or experimental error with zero mean and common

variance.

MANAGEMENT OF THE EXPERIMENTS

CHARACTERISTICS OF THE SUGARCANE BAGASSE SILAGE

PROCESSING SITE

The study was conducted between the months of January and May 2019

(harvest time), a shed was available that met the conditions and

characteristics to develop the Ammonification process such as:

protected from direct solar radiation, rodents and with a dry

ijk

Effect of different study factors on the in situ ruminal degradability of ammonified sugarcane

bagasse.

environment that favored the integral maintenance of the study

material; close to the milling equipment to develop the process of sugar

cane bagasse.

OBTAINING BAGASSE FROM SUGAR CANE

The sugarcane crop was selected with an age of six (6) months, material

that was harvested, collected and transported to the sugarcane mill for

the extraction of sugarcane juice; as a result of this process, sugarcane

bagasse was obtained and used as vegetative material.

AMMONIFICATION PROCESS WITH UREA

The nitrogen source used was agricultural urea (46% N), the

ammonification procedure consisted of filling the 20 kg containers with

the bagasse, then compacting it and moistening it in the urea-water

solution until the containers were full, the containers were covered and

hermetically sealed with packing tape to guarantee an anaerobic

environment.

The arrangement of the treatments, resulting from the combination of

the study factors, is as follows:

Percentage of Urea

Fermentation times (Days)

Urea 0% Urea

Urea 3%.

Urea 5%.

After the first 15 days, the containers were opened with their respective

treatments and days of fermentation, which allowed the ammoniated

material to be ventilated, guaranteeing the excessive elimination of

NH3.

The samples were obtained with their respective doses of urea and

fermentation times, and the drying and milling process was carried out.

One kg of ammonified sugarcane bagasse, per treatment, was

dehydrated for 72 hours, at temperatures between 45 and 60°, by means

of an artisanal machine belonging to the Production Department of the

April - June vol. 2. Num. 2 - 2023

Faculty of Veterinary Sciences. Once the aforementioned process was

completed, the weights of each sample were obtained to determine the

moisture value.

Materials and methods

After drying the sample, it was ground by means of a special blade mill

with sieves of 1 mm diameter for bromatological analysis and 2 mm for

degradation. Subsequently, the ground samples were placed in plastic

tetra pack bags duly labeled with date, quantity and characteristics of

the sample.

Once the ground treatments were obtained with their respective

treatments with the fermentation days, 1 kg of each of them were

weighed for the respective bromatological analysis.

Effect of urea level and different fermentation times on in situ ruminal

degradability of sugarcane bagasse dry matter.

The present experiment was carried out in the Department of Animal

Production of the Faculty of Veterinary Science of the Technical

University of Manabí, located in the Santa Ana canton, parish of

Lodana, geographically located in the east center of the province of

Manabí, at 1° 12'latitude South and 80° 22¨ longitude West. Its altitude

is 50 meters above sea level and its highest point reaches a height of

400 meters above sea level.

In Santa Ana, the rainy season is very hot, oppressive and cloudy and

the dry season is hot, sultry and partly cloudy. During the course of the

year, the temperature generally varies from 20 °C to 30 °C and rarely

drops below 19 °C or rises above 32 °C (ECURED, 2019).

The average temperature is 20.2 ºC, with an average annual

precipitation of 4222.7 mm, with the months of August y September

the driest and June y July the rainiest (ECURED, 2019).

The time taken for this research was about three (3) months in the field

(February - April).

The research was carried out at the animal production department of the

Technical University of Manabí, Faculty of Veterinary Sciences,

located in the Santa Ana canton, in the Lodana parish.

Effect of different study factors on the in situ ruminal degradability of ammonified sugarcane

bagasse.

Fistulated crossbred cattle were used for a period of 15 consecutive days

with a rest of one day for the inclusion of the samples.

In an analytical balance, 2.5 g of sample were weighed, ground to 2

mm, and each one was placed in a nylon bag previously labeled, which

were sealed with plastic ties, of each treatment to be analyzed, with

three repetitions, for its location in the rumen. Subsequently, they were

extracted from the rumen at 0, 3, 6, 12, 24, 48 and 72 hours,

respectively, with the different samples taken. Then, they were placed

in a cooler with ice, the cannula was closed, and the surroundings of the

cannula were washed with an iodine solution and with applications of

silver spray (bactrovet) to avoid contamination.

The samples were taken to the freezer and incubated. After 72 hours,

they were removed from the freezer and washed several times in a

washing machine until the residues were eliminated and the water was

colorless; this process was carried out for each of the hours and

treatments to be evaluated.

All the bags for the zero hour (0) were not introduced in the animal,

they were only included at the moment of washing the samples.

Subsequently, the samples were centrifuged, the moorings were

removed and they were taken to the oven for 24 hours at 105 °C. After

24 hours, the samples were weighed, then it was verified how much

they degraded in each time.

For the processing of the samples, the analysis used by (Orskov and

McDonald 1979) was used. The calculation of degradability was

carried out using the following formula:

Degradabilidad=

Mpre - Mpost

Mpre

!!𝑋!100

Where:

%DISMS: Percentage of in situ DM degradation.

Mpre: Pre-incubated matter

Mpost: Post-incubated matter

April - June vol. 2. Num. 2 - 2023

The data were analyzed through an Analysis of Variance. Differences

between the study and interaction factors were observed through

Tukey's Test at 5% probability, using the SAS statistical package

(2013). Descriptive statistics were also performed, taking into account

the mean (central tendency), standard deviation and coefficient of

variation (dispersion measures). The results were represented in tables

and graphs, bearing in mind the interest they reflect, according to the

objectives set.

3. Result

In the analysis of variance (Annex 6), a highly significant difference

was observed for the main factor urea (P<.0001), giving the best soluble

fraction value for the ammonification of sugarcane bagasse at 5% urea

(Table 4.3), which was 49.93%. The results obtained are similar to those

found in the study by Castellano et al. (2017)who conclude that the

average DIVMS values present a highly significant difference (P<0.01)

between urea levels. In addition, an increase of 11.17% of DIVMS was

observed when applying 6% urea (66.59%). In contrast, differences

were found for the effect of the origin of the panca. In another study by

Fuentes et al. (2001)reported values of 66.05, 71.50 and 71.94%

DIVMS when treating ground, chopped and whole corn stover,

respectively, with 4% ammonia over a period of 4 weeks in relation to

untreated stover (64.67% DIVMS). This is also corroborated with the

results of the study Tesfaye et al. (2006) observed that treatment with

5% urea increased DIVMS by 7.7%, in proportion to the untreated

pank.

In Figure 4.3, a significant direction change interaction (P> 0.0453) is

highlighted. The improvement in digestibility of highly fibrous

materials treated with urea can be attributed to the effect of ammonia

on the fiber cell walls. The results found in the study by Araiza et al.

(2013)in which it is indicated that the chemical action of ammonia

allows breaking the bonds of the structural complexes of cellulose and

hemicellulose, which favors greater quantity and availability of soluble

carbohydrates for ruminal microorganisms; as well as Reyes

(2018)found that the soluble or rapidly degradable fraction (A) of

ammonified peanut shells achieved values ranging between 50.68 and

53.19 %. The exposed results differ from reported by Araiza et al.

(2013)with higher values in corn and apple silages that were in the order

of 39.49 % and 42.53 %. These results show that the interaction is

Effect of different study factors on the in situ ruminal degradability of ammonified sugarcane

bagasse.

significant because the simple factor urea is affecting the study variable.

Table 2 Effect of different levels of urea 3 and 5% on in situ ruminal

degradability (%) of dry matter (DM) of ammonified sugarcane

bagasse.

UREA EFFECT

Parameters

Pr > F

FS (%)

13,22c±3,13

44,57b±3,13

49,93a±3,13

<.0001

FI (%)

37,90a±1,36

26,04b±1,46

26,59b±0,71

<0.0045

TD (%)

0,04a±1,32

0,04a±1,08

0,03a±4,19

<0.7757

DE 2%.

3,93c±3,93

61,61b±5,56

65,17a±7,01

<.0001

DE 5%.

30,48c±4,82

55,95b±1,44

59,76a±0,98

<.0001

DE 8%.

26,28c±4,87

53,15b±1,54

57,25a±0,45

<.0001

FS: Soluble fraction; FI: Insoluble but potentially degradable fraction;

TD: Degradability rate; DE: Effective degradability.

a,b,c Different letters in the column differ statistically at 5 % (Tukey).

Table 3. Effect of different ammonification times (30 and 44) on in situ

ruminal degradability (%) of dry matter (DM) of ammonified

sugarcane bagasse.

EFFECT OF AMMONIFICATION TIME

Parameters

Pr > F

FS (%)

35,11a ± 6.18

36,71a ± 5.39

0.1963

FI (%)

29,04a ± 2.93

31,32a ± 2.66

0.3999

TD (%)

0,04a ± 2.57

0,031b ± 2.80

0.0051

DE 2%.

55,73a ± 4.14

54,74b ± 6.42

0.0507

DE 5%.

49,38a ± 5.12

48,08b ± 4.68

0.0162

DE 8%.

46,06a ± 4.80

45,05a ± 6.73

0.1118

FS: Soluble fraction; FI: Insoluble but potentially degradable fraction;

TD: Degradability rate; DE: Effective degradability.

a,b,c Different letters in the column differ statistically at 5 % (Tukey).

April - June vol. 2. Num. 2 - 2023

The insoluble but potentially degradable fraction was highly significant

in the analysis of variance (P 0.0045) only for the simple factor urea

(Annex 7). Urea at 3% and 5% were statistically equal with values of

26.04 and 26.59, respectively, demonstrating a numerical difference

(Table 4.3).

In the analysis of variance, a highly significant difference was observed

for the study factor time (Annex 8), with the lowest degradability rate

(day 44) with a value of 0.031% (Table 4.3). These results contrast with

those of Reyes (2018)who corroborates that degradation rates lower

than 0.02*h-1 are characteristic of low quality feeds that need more

time in the rumen for degradation. Another study that differs from the

results obtained are those of Araiza et al. (2013)which shows that

degradation rates are higher than those reported for corn silage with

0.039*h-1, except in treatment four with 9% urea, which obtained

0.02*h-1.

Effective degradability at 2, 5 and 8%.

Effective degradability at 2%.

The analysis of variance (Annex 9) for this variable was highly

significant for the simple urea effect (P 0.0001), as was the interaction

with (P 0.0001). The analysis of variance (Annex 11) for this variable

was highly significant for the simple urea effect (P 0.0001), as was the

interaction with (P 0.0002).

These results are similar to those reported by Luna. (2017)for whom

chemical treatment with different levels of urea improved digestibility

using 4 % ammonia in corn stover and wheat bran. Similarly, Saadullah

et al. (1980) and Fondevila et al. (1994)(1994) corroborate the above,

reporting that the treatment of straw with 3 ± 5 % urea increased dry

matter digestibility by 11 ± 15 %, which could be attributed to the

decrease in neutral detergent fiber content.

4. Conclusions

Based on the literature consulted and the results obtained in the present

bromatological study, concerning the operational aspects to evaluate the

in situ degradation of ammonified sugarcane bagasse silage to supplement

the diet of cattle in times of drought, the following conclusions can be

Effect of different study factors on the in situ ruminal degradability of ammonified sugarcane

bagasse.

drawn:

The rumen degradation rate remained constant at all urea inclusion

levels, which is very important in ruminant feeding, taking into

consideration that high degradability rates can cause digestive

problems.

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