The Effect of Clove Essential Oil Treatment on the Cell Wall Components of Wheat Straw

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Journal of Agricultural Science and Technology A 7 (2017) 68-72
doi: 10.17265/2161-6256/2017.01.010
The Effect of Clove Essential Oil Treatment on the Cell
Wall Components of Wheat Straw
Hülya Özelçam1, Sema Özüretmen1, Hasan Hüseyin İpçak1 and Aylin Dereboylu2
1. Department of Animal Science, Faculty of Agricultural, Ege University, İzmir 35100, Turkey
2. Department of Biology, Faculty of Science, Ege University, İzmir 35100, Turkey
Abstract: The aim of this study was to determine the effects of additions of different doses of clove oil (Syzygium aromaticum L.) on
cell wall component of wheat straw. For this purpose, wheat straw was treated with 100 ppm and 200 ppm clove oil and applied at
two different time period (1 h and 5 h). The microscopic analysis was made on cell wall components of untreated and treated of the
straw. According to the research findings, with increasing doses and time of clove oil treatment, particularly, neutral detergent fiber
(NDF) and acid detergent fiber (ADF) content of straw significantly (P < 0.05) reduced, approximately at the level of 15% for NDF
and 13% for ADF, respectively. The lowest NDF, ADF, acid detergent lignin (ADL) and cellulose contents were found in 200 ppm
dose and 5 h period. However, the lowest stem section thickness likewise was determined in 5 h period (P < 0.05), but there was no
significant difference between the dose. Consequently, it could be said that the addition of clove oil have a positive influence on cell
wall components and stem section thickness of wheat straw.
Key words: Wheat straw, clove oil, cell wall components.
1. Introduction
Structural carbohydrates within the structure of
straws are essentially of lignocellulosic structure
(30%-50% cellulose + 25%-30% hemicellulose +
10%-20% lignin) [1]. The structure expressing cell
walls of feeds is named as neutral detergent fiber
(NDF = hemicelluloses + cellulose + lignin), and it is
particularly important in the preparation of dairy cattle
rations. As a matter of fact, NDF is tightly correlated
with feed intake, rumination and chewing time [2].
Cellulose and lignin content of feeds are designated as
acid detergent fiber (ADF) and its low level is desired
in the feed. The acid detergent lignin (ADL) can form
strong chemical bonds with the cellulose and
hemicellulose of the feed, and lignocellulosic structure
is generally regarded as indigestible. Thus, the
lignocellulosic structure shows resistance against
microbial enzymes during in the rumen fermentation,
thereby decreasing the digestibility of the straw [3].
Corresponding author: Hülya Özelçam, professor, research
fields: forages, silage, ruminant nutrition, nylon bag technique.
So far, attempts have been made to degrade the
bonds of lignocellulosic structure within these wheat
straws and to enhance the digestibility level of the
straws. To that end, various physical, chemical [4-6]
and microbiological [7] studies have been performed
[8, 9]. Thus, there can be seen the thickened cell wall
structure in the unprocessed wheat straw, while
polysaccharides are decomposed a little in steam
treatment, and the bonds between hemicellulose and
lignin are degraded in alcohol treatment [10]. On the
other hand, it has been revealed in the researches that
etheric oils have effects on the ruminal digestibility of
the feed [11-13]. The reason is that the etheric oils
have a fetal effect on bacteria with its special effect
mechanism. This mechanism impairs the electron, ion
or K balance within the cell wall, causing shrinking
and death of bacteria via effusing the cell content out
[14, 15].
Furthermore, it is reported that this effect of etheric
oils is increased or decreased in accordance with time
[16, 17]. The effect left on the dead cell tissue by the
etheric oils has not yet been observed. However, it has
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The Effect of Clove Essential Oil Treatment on the Cell Wall
Components of Wheat Straw
69
been revealed that upon clove oil addition to the wheat
straw, in vitro digestibility and metabolizable energy
value of the feed are increased [18]. In fact, clove oil
(Syzygium aromaticum L.) is an etheric oil type
having strong antimicrobial, antifungal and
therapeutic features with the main component eugenol
[19-22].
The aim of this study was to reveal the effect of
treatment with clove oil in different dose and time
periods on the cell wall components of wheat straw.
2. Materials and Methods
The materials of the study consist of wheat straw
and clove oil with 66.59% eugenol. A total of four
treatment groups including one control (without clove
oil addition) were formed. Wheat straws (1,000 g)
were treated with 100 ppm and 200 ppm clove oil and
at 1 h and 5 h time periods. Clove oil was sprayed on
wheat straw, which was weighed into the polyethylene
bags by a spray bottle. During the treatment, the
straws were kept in dry and dark environment. At the
end of the treatment, the straws were ground in 1 mm
screen, and the cell wall components were determined.
The NDF, ADF and ADL components of the straws
were determined in accordance with Van Soest
analysis method. Moreover, the hemicellulose
contents of the straws were calculated from
NDF-ADF difference and cellulose contents from
ADF-ADL difference [23], as Eqs. (1) and (2):
Hemicellulose% = NDF% – ADF% (1)
Cellulose% = ADF% – ADL% (2)
Furthermore, microscopic analyses of the straws
(Olympus bx-51, C-5050) were performed, and the
sectional thickness of cell wall was determined as well.
Image-Pro Express image analysis program has been
used in the calculation of the cell wall sectional
thicknesses. Analyses were conducted in two different
days. In each day, four replicates for each sample
were studied.
SPSS (SPSS version 21) package program
(multivariate analysis of variance (MANOVA)) has
been used in statistical evaluation of the data obtained
from the study. In comparison of the differences
between the mean values, Duncan multiple
comparison test (P < 0.05) has been used [24].
3. Results and Discussion
In the study, the effect of clove oil on the wheat
straw cell wall components on different dose and time
periods has been investigated. As known, cell wall is a
complex structure made of hemicellulose, cellulose
and lignin. Particularly lignin can form strong bonds
with hemicellulose and cellulose on the cell wall, thus,
decreasing the digestibility of the straws. So, lignin
has to be degraded for the digestibility in the rumen to
occur. However, rumen microorganisms are
inadequate in degrading this hard structure [25]. To
that end, a plurality of delignification studies has been
performed. Nevertheless, there has not been any study
pertaining to positive or negative effect of etheric oils
on cell wall components.
In the study, the effect of clove oil treatment with
wheat straw on cell wall components with different
doses and times is presented in Table 1. Accordingly,
Table 1 Effect of clove oil treatment on cell wall component of wheat straw with different doses and times (in dry matter%).
Clove oil NDF ADF ADL Hemicellulose Cellulose
Control 75.39 ± 1.34a 45.19 ± 0.23a10.18 ± 0.76a30.20 ± 1.26a 35.28 ± 0.89a
100 ppm, 1 h 70.30 ± 0.64
b
42.71 ± 0.49
b
9.39 ± 0.89ab 27.59 ± 0.77ab 33.32 ± 1.12ab
200 ppm, 1 h 65.48 ± 1.04c
d
41.55 ± 0.67
b
8.65 ± 0.43ab 23.93 ± 1.40c 33.47 ± 0.67ab
100 ppm, 5 h 67.16 ± 0.92c 41.46 ± 0.34
b
9.54 ± 0.29ab 24.65 ± 1.04
b
c 31.86 ± 0.74
b
c
200 ppm, 5 h 63.74 ± 0.19
d
39.18 ± 1.22c7.75 ± 0.24
b
24.62 ± 1.00
b
c 30.20 ± 0.19c
P value 0.00 0.00 0.03 0.00 0.01
Dose × time 63.23 ± 0.89 39.02 ± 0.43 8.41 ± 0.26 24.20 ± 0.66 30.61 ± 0.48
P value 0.16 0.20 0.18 0.06 0.97
The differences between means in the same column with different letters are significant (P < 0.05).

The Effect of Clove Essential Oil Treatment on the Cell Wall
Components of Wheat Straw
70
Table 2 Effect of clove oil treatment on sectional thickness of wheat straw with different doses and times.
Clove oil Control 100 ppm, 1 h 200 ppm, 1 h 100 ppm, 5 h 200 ppm, 5 h P value
Sectional thickness (µm) 297.2 ± 6.9
a
200.1 ± 2.6
b
167.9 ± 2.5
c
120.6 ± 5.7
d
110.2 ± 4.0
d
0.00
The differences between means in the same row with different letters are significant (P < 0.05).
(a) General image of cell wall in the wheat straw (b) Untreated wheat straw
(c) Treated with clove oil (100 ppm, 1 h) (d) Treated with clove oil (200 ppm, 1 h)
(e) Treated with clove oil (100 ppm, 5 h) (f) Treated with clove oil (200 ppm, 5 h)
Fig. 1 The light microscopy images of the wheat straw treated with clove oil at different dose and time period.
it has been detected that with increasing of dose and
time, the clove oil has decreased the cell wall
components of wheat straw significantly (P < 0.05).
This decrease in 5 h 200 ppm group, with respect to
the control group, is 15%, 13%, 24%, 21% and 14%
for NDF, ADF, ADL, hemicellulose and cellulose,

The Effect of Clove Essential Oil Treatment on the Cell Wall
Components of Wheat Straw
71
respectively. Moreover, the greatest decrease has been
found in 200 ppm dose and 5 h period, except for
hemicellulose. While, the greatest decrease for
hemicelluloses is found in 200 ppm dose and 1 h
period.
On the other hand, Sun et al. [26] have reported that
6 h treatment of wheat straw with alkaline has enabled
the degradation of ester bonds between ferulic acid,
polysaccharide or p-coumaric acid and lignin within
the cell wall. While, Sirohi and Rai [27] have reported
that treatment of wheat straw with lime has caused
decrease in NDF and ADL content, and the greatest
decrease compared to the control group is in the 3rd
week. Nasehi et al. [9] have shown that NDF, ADF
and ADL rates have decreased in enzyme-treated
wheat straw, while crude protein, and in situ dry
matter and organic matter digestibility have increased.
In the study, the effect of clove oil treatment with
wheat straw on cell wall sectional thickness of straw
with different doses and times is presented in Table 2.
Accordingly, it has been determined that with
increasing of the dose and time, the clove oil has
thinned the cell wall sectional thickness of wheat
straw significantly (P < 0.05). The greatest thinning
has been found in 200 ppm dose and 5 h period (110.2
µm), with respect to the control group (297.2 µm).
Also, it has been seen in the images obtained from
light microscopy (Fig. 1) that cortical parenchyma
cells are swollen in the 1st hour, and partially or
completely decomposed in the 5th hour, and also there
are partial degradations on the epidermis (cuticula)
layer in the 5th hour.
In the present study, it is presented that the clove oil
treatment with wheat straw has significantly decreased
the cell wall components (Table 1) and also thinned
the sectional thickness (Table 2). Actually, it is seen in
the microscopic images that the cells have swollen in
the 1st hour, and there is the explosion of cell within
the middle layer and thinning in the 5th hour. The
situation is considered to be resulted from the entering
of the etheric oil into the cell via diffusion, thereby
increasing the turgor pressure within the cell. Also in
the study, it is seen that the effect of the clove oil has
changed in accordance with dose and time, however,
dose × hour interaction does not matter (Table 1, P >
0.05).
4. Conclusions
Consequently, it can be said that the treatment of
wheat straw with the clove oil has a decreasing effect
on the cell wall components with increasing of the
dose and time. Accordingly, the NDF, ADF, ADL and
cellulose content of wheat straw were observed the
greatest decrease in 200 ppm dose and 5 h.
Microscopic analyses have found compatible with the
chemical analysis results. But it must be examined by
microscope, such as scanning electron microscope
(SEM) to see more detail about the effect of clove oil
on cell wall components of wheat straw. In vivo
studies are required to see the clove oil treatment of
wheat straw on the animals.
Acknowledgments
The authors would like to thank academician Assoc.
Prof. Dr. Yiğit Uyanikgil and the Department of
Histology and Embryology, Faculty of Medicine, Ege
University for their contributions to the present study.
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