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160
Scientific Journal of Animal Science (2013) 2(6) 160-167
ISSN 2322-1704
Effect of biological treatment on chemical composition and in situ ruminal
degradability of Soybean and Canola straw in Sheep
M.Nasehi
*
, N.M. Torbatinejad, S. Zerehdaran, A.R. Safaei
Department of Ruminant and Poultry nutrition, Faculty of Animal Science, Gorgan University of Agricultural
Sciences and Natural Resources, Gorgan, Iran.
*
Corresponding author; MSc graduate, Department of Ruminant and Poultry Nutrition, Faculty of Animal Science, Gorgan University of
Agricultural Sciences and Natural Resources, Gorgan, Iran.
A R T I C L E I N F O
Article history:
Received 01 June 2013
Accepted 11 June 2013
Available online 29 June 2013
Keywords:
Fungi
Nylon bag
Sheep
Straw
A B S T R A C T
This study was conducted in order to investigate the effect of
biological treatment with Pleurotus florida fungi on chemical
composition and rumen dry matter (DM) and organic matter (OM)
degradability of Soybean and Canola straw. Fungi cultivation
significantly decreased the amount of DM in treatments (P<0.05).
Ash and crude protein (CP) content significantly increased with
processing by fungi in treatments (P<0.01). The amount of Neutral
detergent fiber (NDF) significantly decreased by fungi cultivation
(P<0.05). Ether extract (EE), Acid detergent fiber (ADF) and Acid
detergent lignin (ADL) content of treatment did not significant
difference with processing by fungi. The soluble fraction (a) and
potential degradability (a+b) of DM degradability significantly
increased with processing in treatments (P<0.01). The
insoluble but
potentially degradable fraction (b) of DM degradability significantly
increased with processing by fungi in Soybean straw (P<0.01) but did
not significant difference in Canola straw. The (a) fraction of Canola
straw OM degradability significantly increased by processing (P<0.05)
but did not significant difference in Soybean straw. The (b) fraction of
Soybean straw OM degradability significantly increased by processing
(P<0.05) but did not significant difference in Canola straw. The (a+b)
fraction of OM degradability significantly increased with processing in
treatments (P<0.05). This research results generally reflect
improvement in nutritive value of Soybean and Canola straw with
Original article
Contents lists available at Sjournals
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M.Nasehi et al. / Scientific Journal of Animal Science (2013) 2(6) 160-167
161
treatment by fungi in ruminant nutrition.
© 2013 Sjournals. All rights reserved.
1. Introduction
There are a lot of agricultural residues (crop residues and agro industrial residues) annually produced in
agricultural countries all over the world of which Iran is no exception. The large portion of these residues are
important feed stuff for ruminants and can be used as a potentially important source of carbohydrates and energy;
however, the utilization of these materials as a feed source for ruminants are limited for their complex biological
structure and low protein content (Rodrigues et al., 2008; Yalchi and Hajieghrari., 2011). Lignocelluloses is the
major component of crop residue cell wall, especially secondary cell wall, with cellulose, hemicelluloses and lignin
content in which lignin inheres in the cellulose and hemicelluloses matrix. The low ability of lignocelluloses to
hydrolyze (more for crystalline structure of cellulose fibrils and presence of lignin) reduces the digestibility and
restricts efficient utilization of the feed produced by ruminal microorganisms. Although, microorganisms within the
rumen are able to exude enzymes that have potential to directly hydrolyze cellulose and hemicelluloses in the
rumen. The complex network formed by cellulose, hemicelluloses and lignin reduces their digestibility because of
lacking ligninolytic activity (Falcon et al., 1995; Otjen et al., 1987; Zadrazil et al., 1985). Therefore, they are not
very efficient in order to break down the lignocellulosic bond of straw. The various methods that could increase its
nutritive value, physical and chemical processing have been studied (Matsuzaki et al., 1994; Rahal et al., 1997).
Although these methods have advantages, they are costly, low in effectiveness, not environmentally friendly and
also require application of technology (Leng, 1991; Sharma et al., 1993). These factors limit their application,
particularly at small farm levels. Recently, biological delignification of straw by solidstate fermentation (SSF) has
been considered because of its capacity to remove lignin preferentially (Moyson and Verachtert, 1991). Fungal
treatment could be an approach to convert low quality crop residues into a higher quality of ruminant feed (Arora
et al., 1994; Zadrazil et al., 1997). Attempts had been made to identify species of white-rot fungi for their ability to
grow on straws that improved their nutritive value (Yamakawa et al., 1992). During the SSF of wheat straw by
fungi, its OM and detergent fibre content could be reduced and the lignin selectively removed from the
lignocellulosic complex (Singh et al., 1990; Kundu, 1994). The CP and Ash were also increased in the treated straw
(Moyson and Verachtert, 1991). Such changes were dependent on the strain of fungi and the cultural conditions
(Tripathi and Yadav, 1991). Among the edible white-rot fungi, the Pleurotus species have been shown to be more
efficient (Zadrazil et al., 1996). The potential of some species of Pleurotus fungi such as P.ostreatus and P. eryngii
to reduce indigestible cell wall components and increase dry mater digestibility (DMD) of straw has been reported
(Agosin et al., 1986; Singh et al., 1990). Therefor this study conducted in order to investigate the effect of biological
treatment with Pleurotus florida fungi on chemical composition and rumen DM and OM degradability of Soybean
and Canola straw.
2. Materials and methods
2.1. Treatment of straws and chemical analyze
Soybean and Canola straw were collected from local farms in Gorgan, Golestan province, Iran. Pleurotus
florida fungi were used for samples treatment. Soybean and Canola straw treatment was carried out in 1000 ml
bottles. 50 g of each straw was put in individual bottle and water added to give moisture content of about 85%.
The bottles autoclaved at 121°C for 20 min. Each bottle was inoculated with 5% (w/w) spawns of Pleurotus florida
fungi (Jahromi et al, 2010). Each treatment was four replicate. The bottles incubated in a incubator where
temperature was automatically adjusted to 25°C and relative humidity was kept at 78±5%. After 21 days samples
were dried in oven (60
0
c) in order to stop fungi growth and then chemical composition, rumen DM and OM
degradability were determined.
DM determined by drying the samples at 105°C overnight and Ash by igniting the samples in muffle furnace
at 550°C for 8 h. EE of the samples were determined by soxhlet extraction method (AOAC, 2005). NDF, ADF and
M.Nasehi et al. / Scientific Journal of Animal Science (2013) 2(6) 160-167
162
ADL content was measured by Fiber-Tec system (Vansoest et al, 1991). Nitrogen (N) content was measured by the
Kjeldahl method and CP was calculated as N*6.25(AOAC, 2005).
2.2. In situ degradation procedures
Two ruminally cannulated Dalagh rams (about 45 kg BW) were used to determine in situ degradation
characteristics. Rams were housed in individual tie stalls bedded with sawdust. Rams fed diets containing wheat
straw (70%) and concentrate mixture (30%) at the maintenance levels (NRC, 1985). The concentrate mixture
contained barley grain and wheat bran.
Dacron bags (40-45 micron pore size) were filled with 3 g dried and ground samples then incubated in the
rumen of rams for the periods of 0, 4, 8, 12, 24, 48, 72 and 96h. Bags were removed, washed and dried according
to the procedure of Ørskov et al. (1980). Rumen degradation kinetics of DM and OM was fitted by the nonlinear
model proposed by Ørskov and McDonald (1979) using Fitcurve software version 6 (Chen, 1995).
P = a + b (1-e
-ct
)
Where:
P = Percentage of degradability for response variables at t.
t = Time relative to incubation (h)
a = soluble fraction (%)
b = insoluble but potentially degradable fraction (%)
c = Rate constant for degradation (h
-1
)
e = 2.7182 (Natural logarithm base)
Following determination of these parameters, the effective degradability of DM and OM in the samples was
calculated using and equation described by Ørskov and McDonald (1979):
ED = a + (bc)/(c+k)
Where:
ED = Effective degradability for response variables (%)
a = soluble fraction (%)
b = insoluble but potentially degradable fraction (%)
c = Rate constant for degradation (h
-1
)
k = Rate constant of passage (h
-1
)
Statistical Analysis
For all data, a completely randomized design with a 2 × 2 factorial arrangement were used. The experimental
factors were straw type at 2 levels (Soybean and Canola straw) and fungus application at 2 levels (control and
Pleurotus florida fungi). Each parameter was measured with four replicate. the obtained data were analyzed for
parametric statistics, including analysis of variance using the general linear model procedure (GLM) and the
differences among treatments’ means were compared by Tukey’s test. SAS statistical software (version 9.1) were
used for statistical analyzes.
Tabale 1
The effect of processing on chemical compositions (%) of experimental treatments.
Items
USS
TSS
UCS
TCS
P value
SEM
DM
88
b
81.38
c
89.32
a
83.40
d
0.04
0.14
Ash
3.48
d
5.46
c
12.81
b
14.22
a
0.0001
0.33
EE
1
a
0.82
a
0.79
a
0.76
a
0.3
0.07
CP
4.36
d
5.37
c
6.63
b
8.48
a
0.009
0.12
NDF
71.42
a
63.83
b
64.89
b
61.02
c
0.01
0.61
ADF
61.87
a
61.69
a
58.73
b
58.02
b
0.7
0.75
ADL
10.22
bc
9.80
c
14.65
a
13.69
ab
0.7
0.91
USS= Untreated Soybean straw, TSS= Treated soybean straw, UCS= Untreated Canola straw, TCS= Treated
Canola straw, P value= Probability value, SEM= Standard error of means; DM= Dry matter, EE= Ether extract, CP=
Crude protein, NDF= Neutral detergent fiber, ADF= Acid detergent fiber, ADL= Acid detergent lignin. Means in the
same row with the different superscript are significantly different {(P<0.01) and (P<0.05)}.
M.Nasehi et al. / Scientific Journal of Animal Science (2013) 2(6) 160-167
163
3. Results
3.1. Chemical composition
Chemical composition of experimental treatments is presented in Table 1. Fungal treatment significantly
(P<0.05) decreased the DM content of straws, among the treatments, DM was higher for untreated canola straw
(89.32%) compared to other treatments. The Ash and CP content of treatments significantly increased with fugal
treatments (P<0.01), but among the treatments, CP was higher for treated Canola straw (8.48%) compared to
other treatments (Table
Fungal treatment had not significant effect on EE content of experimental treatments (P>0.05) (Table 1). The
NDF content of Soybean and Canola straw significantly decreased by fungal treatment (P<0.05). NDF was lower for
treated Canola straw (61.02%) compared to other treatments (Table 1). ADF and ADL content of treatments did
not signifant change by processing with fungi. The ADF content for treated Canola straw (58.02%) was lower
compared to other treatments. The ADL content for treated Soybean straw (9.80%) was lower compared to other
treatments (Table 1).
3.2. In situ degradation
The DM degradability of experimental treatments is presented in Table 2 (Fig 1). Fungal treatment
significantly increased DM degradation at 96h in both straw (P<0.01). The OM degradability of experimental
treatments is presented in Table 3 (Fig 2). Fungal treatment significantly increased OM degradation at 96h in both
straw (P<0.01). At most of the rumen incubation periods, the degradability of DM and OM was higher for the
treated Canola straw than the other treatments (Table 2 and Table 3) . Table 4 shows the degradability parameters
of the DM obtained from the fitted values. The soluble fraction (a) and potential degradability (a+b) of DM
degradability significantly increased with processing in treatments (P<0.01). The
insoluble but potentially
degradable fraction (b) of DM degradability significantly increased with processing by fungi in Soybean straw
(P<0.01) but it did not significant difference in Canola straw. C fraction of DM degradability did not significant
change with fungal treatment in Canola straw, but fungal treatment significantly decreased c fraction of DM
degradability in Soybean straw (P<0.01). DM Effective degradability of experimental treatments significantly
increased with fungal treatment in 2% out flow rate (P<0.05). Fungal treatment did not significant change at 5%
out flow rate of Soybean straw DM degradability, but significantly increased the amount of DM effective
degradability of Canola straw at 5% out flow rate (P<0.05).
Table 2
Ruminal Dry matter degradation (%) of treatments at different incubation times.
Incubation time (h)
SS
TSS
CS
TCS
P value
SEM
0
17.72
c
19.47
b
19.85
b
21.97
a
0.0008
0.72
4
21.97
c
26.42
a
22.72
c
25.05
b
0.0001
0.54
8
27.25
b
29.22
ab
28.72
ab
31.52
a
0.05
1.37
12
29.55
c
31.97
b
33.02
b
38.57
a
0.0001
1.09
24
39.30
c
36.40
d
41.72
b
44.27
a
0.0001
0.85
48
43.07
b
42.52
b
46.25
b
50.02
a
0.003
1.70
72
46.85
c
50.02
b
48.40
bc
53.05
a
0.0001
0.87
96
47.57
c
55.82
a
49.40
b
55.25
a
0.0001
0.73
SS= Soybean straw, TSS= Treated soybean straw, CS= Canola straw, TCS= Treated Canola straw; P value=
Probability value, SEM= Standard error of means, Means in the same row with the different superscript are
significantly different {(P<0.01) and (P<0.05)}.
M.Nasehi et al. / Scientific Journal of Animal Science (2013) 2(6) 160-167
164
Table 3
Ruminal Organic matter degradation (%) of treatments at different incubation times.
Incubation time (h)
SS
TSS
CS
TCS
P value
SEM
0
19.40
c
19.86
c
22.42
a
24.39
a
0.0001
0.64
4
21.94
d
27.19
b
25.04
c
38.65
a
0.0001
0.65
8
26.56
c
29.62
b
30.87
b
47.55
a
0.006
1.06
12
28.40
c
33.64
b
34.49
b
50.69
a
0.01
0.71
24
42.45
b
37.03
c
42.61
b
53.73
a
0.0001
1.15
48
44.92
c
43.28
c
47.55
b
55.57
a
0.0001
1.10
72
47.85
c
47.52
c
50.86
b
57.78
a
0.0008
0.85
96
48.72
d
55.27
b
53.44
c
60.50
a
0.0001
0.61
SS= Soybean straw, TSS= Treated soybean straw, CS= Canola straw, TCS= Treated Canola straw; P value= Probability value
,SEM= Standard error of means, Means in the same row with the different superscript are significantly different {(P<0.01)}.
DM Effective degradability of Canola straw increased with fungal treatment in 8% out flow rate (P<0.05), but
fungal treatment did not significant difference in DM effective degradability of Soybean straw at 8% out flow rate.
Table 5 shows the degradability parameters of the OM obtained from the fitted values. The (a) fraction of Canola
straw OM degradability significantly increased by processing (P<0.05), but did not significant difference in Soybean
straw.
Fig. 1. DM degradation of treatments. Fig. 2. OM degradation of treatments.
The (b) fraction of Soybean straw OM degradability significantly increased by processing (P<0.05), but did not
significant difference in Canola straw. The (a+b) fraction of OM degradability significantly increased with
processing in treatments (P<0.05).
C fraction of OM degradability did not significant change with treatment in Soybean straw, but fungal
treatment significantly increased this fraction in Canola straw (P<0.01). Biological treatment with fungi significantly
increased the amount of OM effective degradability at 2% out flow rate (P<0.01). OM effective degradability of
Soybean straw at 5% and 8% out flow rates did not significant change with fungal treatment, but OM effective
degradability of Canola straw at these out flow rates significantly increased with fungal treatment (P<0.01). (Table
5).
M.Nasehi et al. / Scientific Journal of Animal Science (2013) 2(6) 160-167
165
Table 4
Ruminal Dry matter degradation parameters and effective degradability of treatments.
Items
SS
TSS
CS
TCS
P value
SEM
a (%)
17.40
b
22.67
a
18.65
b
21.05
a
0.007
0.45
b (%)
30.72
b
40.05
a
30.70
b
33.32
b
0.007
1.04
a+b (%)
48.12
c
62.70
a
49.40
c
54.37
b
0.001
1.16
c (/h)
0.04
a
0.01
b
0.05
a
0.05
a
0.0005
0.003
ED= 0.02
38.60
c
40.92
b
40.30
b
44.95
a
0.01
0.39
ED=0.05
32
c
32.80
bc
34.35
b
38.82
a
0.01
0.49
ED=0.08
28.57
c
29.67
bc
30.80
b
34
a
0.04
0.47
SS= Soybean straw, TSS= Treated soybean straw, CS= Canola straw, TCS= Treated Canola straw; a= Soluble fraction,
b= insoluble but potentially degradable fraction, a+b= potential degradability, c= rate of degradation of fraction b (h-
1), ED= effective degradability in out flow rates (0.02, 0.05, 0.08) h-1, P value= Probability value ,SEM= Standard error
of means, Means in the same row with the different superscript are significantly different {(P<0.01)}.
Table 5
Ruminal Organic matter degradation parameters and effective degradability of treatments.
Items
SS
TSS
CS
TCS
P value
SEM
a (%)
17.69
a
23.12
ab
21.75
b
24.66
a
0.03
0.53
b (%)
31.57
b
35.12
a
31.42
b
32.71
ab
0.1
0.81
a+b (%)
49.26
c
58.25
a
53.18
b
57.37
a
0.02
0.95
c (/h)
0.04
b
0.02
b
0.04
b
0.12
a
0.001
0.01
ED= 0.02
39.65
d
40.85
c
42.87
b
53.20
a
0.0001
0.35
ED=0.05
32.80
c
33.45
c
36
b
48.62
a
0.0001
0.39
ED=0.08
29.22
c
30.40
c
32.47
b
45.32
a
0.0001
0.38
SS= Soybean straw, TSS= Treated soybean straw, CS= Canola straw, TCS= Treated Canola straw; a= Soluble fraction,
b= insoluble but potentially degradable fraction, a+b= potential degradability, c= rate of degradation of fraction b (h-
1), ED= effective degradability in out flow rates (0.02, 0.05, 0.08) h-1, P value= Probability value ,SEM= Standard error
of means, Means in the same row with the different superscript are significantly different {(P<0.01) and (P<0.05)}.
4. Discussion
4.1. Chemical composition
The protein content of the mycelium was reported relatively high (Ragunathan et al., 1996), so it was
expected that the treated straw, that contained fungal mycelium to have a higher concentration of CP. An increase
of CP content in wheat straw incubated with Pleurotus species had also been reported (Ardon et al., 1996; Zadrazil
et al., 1996; Fazaeli et al, 2004).
The increase in the CP contents may be due to secretion of certain extra cellular enzymes which are
proteineous in nature into the waste during their breakdown and its subsequent metabolism (Kadiri, 1999;
Akinfemi et al, 2009). CP increase could also be due to the capture of excess nitrogen by aerobic fermentation
(Sallam et al., 2007). suggesting that the treated substrates are good source of protein for livestock. This agrees
with the findings of Zadrazil (1993), Belewu and Okhawere (1998), and Iyayi and Aderolu (2004). The NDF content
of Soybean and Canola straw significantly decreased by fungal treatment (P<0.05) (Table 1). This was due to the
natural habitats of the white-rote fungi that largely depend on organic carbon (for their energy requirement)
including carbon in the form of structural material such as lignocellulosic (Jennings and Lysek, 1999). The losses of
NDF from the straw suggested that these fungi could solubilize and utilize the cell wall as carbon source and thus
changed the ratio of insoluble to soluble carbohydrates in the straw. The decrease in NDF contents of the treated
straw has been supported by other reports (Singh, 1990; Yalchi and Hajieghrari., 2011 ). However, the potential of
NDF degradation among species of fungi could be different (Jalc et al., 1996; Zadrazil et al., 1996).
M.Nasehi et al. / Scientific Journal of Animal Science (2013) 2(6) 160-167
166
4.2. In situ degradability
Fungal treatment significantly increased DM and OM degradation at 96h in both straw (P<0.01) (Table1 and
Table 2). The reason for such improvement in the degradability may be related to the breaking down of cell wall
bonds during the fermentation of straw with the fungi (Jennings and Lysek, 1996; Call and Mücke, 1997; Fazaeli et
al, 2004). These higher values of degradability parameters in fungal treated straws may be explained by the lower
cell wall components in these treatments (Table 4 and Table 5 ). Valmaseda et al. (1991) and Gutierrez et al. (1996)
noted that fermentation of straw with Pleurotus fungi decreased the cell wall components and increased the
soluble fraction of the carbohydrates in the straw that could be as a result of the enzymatic degradation.
5. Conclusion
In conclusion, treatment of Soybean and Canola straw with Pleurotus florida fungi, resulted in a reduction of
those cell wall components and increasing of CP and rumen degradability. The results obtained in this study
suggest that the treatment of Soybean and Canola straw by the application of fungi will help in conversion of
agricultural wastes to higher quality ruminant feed thereby enhancing their digestibility by ruminants. It is
therefore recommended that more work should geared towards this direction to harness the hidden potentials of
agricultural wastes for the benefit of the developing countries.
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