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J.Agric.&Env.Sci.Dam.Univ.,Egypt Vol.11 (2) 2012
EFFECT OF SOME HERBS ON THE RUMEN
FERMENTATION:
1- EFFECT OF GINGER (ZINGIBER OFFICINALE) AND
GARLIC (ALLIUM SATIVUM) ON GAS
PRODUCTION, ENERGY VALUES, ORGANIC
MATTER DIGESTIBILITY AND METHANE
EMISSION, IN VITRO.
TAG EL-DIN
1, A.E.; MOHARAM
1, M.S.; NOUR
1, A.A.; AND NASSER
2, M. E.
A. 1- Animal and poultry production Dept., Faculty of Agirculture, Damanhour University,
Damanhour, Egypt.
2- Animal Production Dept., Faculty of Agriculture, Alexandria University, Alexandria,
Egypt.
ABSTRACT
The present study was designed, to evaluate the effect
of some herbs on gas production, energy values, organic
matter digestibility, microbial protein and methane
production, in vitro. The experimental design was a
completely randomized design in two factors (CRD in 2
factors). An in vitro gas production technique simulate the
rumen fermentation process have been used to evaluate the
potential of feeds to supply nutrients for ruminants and test
the rank of herbs and spices according to their capacity in
lowering of the methane production. Ground samples (100
mg DM) of 30% wheat straw and 70% concentrate were
incubated in 50 ml glass syringes with rumen fluid obtained
from fistulated sheep fed berseem hay and commercial
concentrate mixture (twice a day). The herbs and spices
tested were ginger (Zingiber officinale), garlic cloves (Allium
sativum), and garlic pulp and garlic juice. These spices
(ginger, garlic cloves and garlic pulp) were added at the level
of 0, 0.1, 0.5, 1, 2, and 3% of concentrate while, garlic juice
was added at 0, 0.5, 1, 2, 3, and 5 ml/kg DM. Cumulative gas
production was recorded at 3, 6, 9, 12, 24, 48, 72 and 96 h of
33
J.Agric.&Env.Sci.Dam.Univ.,Egypt Vol.11 (2) 2012
incubation and the kinetics of gas production was described
by using the equation Gas (t)= a + b (1- e-ct
). At 24 h, gas
production volume was the highest value for sample which
contain garlic juice at level (5 ml/kg DM) (P<0.05) and
greater than for other levels of garlic juice (3, 2, 1, and 0.5
mg/kg DM, respectively) and ration which contains ginger at
level (0.1%) (P<0.05) than other rations contained garlic
cloves and garlic pulp sample or control. Total gas
production at 96 h and the maximum rate of gas production
increased when garlic juice was added to samples. Also, the
results showed that the same trend and were significant
differences (p<0.05) and higher in metabolizable energy and
net energy, dry matter digestibility, organic matter
digestibility, short chain fatty acids and microbial protein
for using garlic juice and ginger with different levels than
garlic cloves, garlic pulp, and control. Also, using of garlic
juice and ginger with different levels in the concentrate
reduced methane production with 70-77 % compared with
the control. In an overall conclusion seems that, the
additions of herbs improved the rumen fermentation and
reduce methane production.
Key words: rumen fermentation, gas production, herbs, energy value, microbial
protein, methane production.
INTRODUCTION
In recent years there has been observed an increased interest in
the potential use of herbs and plant essential oils rich in secondary
metabolites to modify rumen fermentation. The goal of such
modifications is to increase the efficiency of the symbiosis between
ruminant and rumen microorganisms in order to improve profits in
animal feeding without negative impact on environment.
Methanogenesis from ruminants is one of the major cause of global
warming and methanogenesis reduces the efficiency of nutrient
utilization hence manipulation of rumen microbial ecosystem for
reducing methane emission is very vital.
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J.Agric.&Env.Sci.Dam.Univ.,Egypt Vol.11 (2) 2012
Modification of ruminal fermentation using feed additives, such
as antibiotics, has proved to be a useful strategy to improve production
efficiency in dairy cattle. The use of antibiotics as feed additives has
proved to be a useful tool to reduce energy and nitrogen losses from
the diet (McGuffey et al., 2001). However, the use of antibiotics as
feed additives in dairy cows has been of increasing concern due to the
potential appearance of residues in milk. Furthermore, the use of
antibiotics as a feed additive has been banned in the European Union
(Russell and Houlihan2003).
Also, recent legislation (1831/2003; EC, 2003) has been
introduced within the European Union to prohibit the use of growth-
promoting antibiotics in animal feeds. The scientific basis for these
restrictions, based around concerns that the use of antibiotics in
animal agriculture can give rise to transmissible resistance factors that
may compromise the therapeutic use of antibiotic in humans, may be
questionable (Casewell et al., 2003). Nevertheless the removal of
antibiotic growth-promoters has led to an increased interest in
alternative means of manipulating rumen fermentation. Here it will
consider one of possible alternatives: natural plant products. For this
reason, scientists are interested in evaluating the potential use of
natural antimicrobials such as herbs and plant extracts. Currently, the
use of plant herbs has resulted in improving rumen ecology (Kamra,
2005; and Wanapat et al., 2008a).
Herbs, spices and their extracts were already used thousands of
years ago in Mesopotamia, Egypt, India, China and old Greece, where
they were appreciated for their specific aroma and various medicinal
properties (Greathead, 2003). When discussing the use of herbs and
spices as feed additives, it can hardly rely only on old believes about
health impact of certain herbs and spices or their active components.
On the other hand, the ruminal methane production is a by-
product of the microbial digestive process and represents a loss of 2–
12% of the feed energy. Furthermore, emission of methane is
considered as one of the most important global environmental issues
(IPCC 2001). Therefore, decreasing methane production is desirable
for reducing the greenhouse gas emission with improved efficiency of
the digested energy utilization (Johnson and Johnson 1995). A
previous report by Kurihara et al. (1999) indicated that methane
33
J.Agric.&Env.Sci.Dam.Univ.,Egypt Vol.11 (2) 2012
energy loss in cattle fed on tropical forage diets was higher than in
those fed on temperate forage diets, due to the relative high levels of
fibre and lignin and a low level of non-fibre carbohydrate in tropical
forages. Also, the livestock in developing countries are
predominantly maintained on a high-roughage diet with little or no
concentrate resulting in increased ruminal methanogenesis. Therefore,
the use of browse species containing secondary compounds as feed
supplement rich in plant secondary metabolites (PSM) for ruminants
in many parts of the tropics is increasing in order to improve animal
performance and reduce methane production (Abdulrazak et al. 2000
and Patra, and Saxena,2010). Tannins and saponins constitute the
major classes of PSM that are currently under research in a number of
laboratories. The antimicrobial action and its effects on rumen
fermentation of these compounds depend on their nature, activity and
concentration in a plant or plant product.
Garlic (Allium sativum) is a herb or spice plant that has been
used by humans as a source of antimicrobial agents for the
gastrointestinal tract. It has a complex mixture of many secondary
plant products including allicin (C6H10S2O), diallyl sulfide (C6H10S),
dialyl disulfide (C6H10S2) and allyl mercaptan (C3H6S) among others
(Lawson, 1996). These compounds could manipulate rumen
fermentation such as decreased in the proportion of acetate and
increased in proportion of propionate and butyrate, inhibition of
methanogenesis and decreased in the CH4: VFA ratio (Busquet et al.,
2005b).
Ginger (Zingiber officinale) has been shown to have
antithrombitic, antioxidant, anti-inflammatory, and anti-bacterial
properties. In the 1970s ginger was first found to have anti-
inflammatory properties including inhibition of prostaglandin
synthesis (Kiuchi et al., 1982).
The objectives of this study are develop new plants or herbs as
dietary supplements for animals to replace chemical additives and
decrease the environmental pollution by reduce methane emission.
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J.Agric.&Env.Sci.Dam.Univ.,Egypt Vol.11 (2) 2012
MATERIALS AND METHODS
Animals and feeds:
Three fistulated Rahmany rams (Egyptian native breed), fed
wheat straw and commercial concentrate mixture twice (30:70), a day
were used for rumen liquor collection in order to application in gas
production technique.
The experimental samples were composed of (70% concentrate
mixture + 30% wheat straw) with different levels of ginger (Zingiber
officinale rhizomes), garlic (Allium sativum bulbs), and garlic residue
and garlic juice were added at the level of 0, 0.1, 0.5, 1, 2, and 3% of
concentrate while, garlic juice was added at the level of 0, 0.5, 1, 2, 3,
and 5 ml/kg DM.
Prepare Garlic cloves:
Garlic cloves are cut to small pieces, dried on 60oC, and then
grinding, after that stored until used.
Prepare garlic juice:
Add 4 liters of distilled water to 1 kg garlic cloves and then
mixing in a blender. The resulting mixture was liquidated confused by
a refinery with a diameter of 1 mm and then tired juice product in
bottles and kept them on the refrigerator 4-5 oC until use.
Prepare garlic pulp:
The pulp resulting from squeezed was dried at 60oC and kept till use.
Four treatments: 1= CFM1 (supplemented with ginger) + wheat
straw (WS), 2= CFM2 (supplemented with Garlic root) +(WS), 3=
CFM3 (supplemented with Garlic dregs ) +(WS), 4= CFM4
(supplemented with garlic juice) +(WS) were used.
Proximate analysis:
Samples were milled through a 1 mm sieve for chemical
analysis and gas production technique. Dry matter (DM) was
determined by drying the samples at hot oven on 80oC overnight and
ash by igniting the samples in muffle furnace at 600oC for 2 h.
Organic matter (OM), crude protein (CP), ether extract (EE), nitrogen
free extract (NFE) and crude fiber (CF) were determined according to
the procedure of AOAC (1990). Chemical analysis was carried out in
duplicate.
Chemical composition of wheat straw, concentrate and herbs
used for in vitro gas production technique is presented in Table (1).
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J.Agric.&Env.Sci.Dam.Univ.,Egypt Vol.11 (2) 2012
In vitro gas production technique
In vitro gas production was undertaken according to the
procedure described by Menke and Steingass (1988).
Syringes preparation Samples (100 mg) of the air-dry feedstuffs were accurately
weighted into 50 ml calibrated glass syringe fitted with plungers. The
buffer solution was used in vitro gas production defined as MB9
(Onodera and Handerson, 1980). The buffer consisted of 2.8 g
NaCl; 0.1 g CaCl2; 0.1 g MgSO4.7H2O; 2.0 g KH2PO4; 6.0 g
Na2HPO4 which dissolved in distilled water and made up to 1 L. The
pH was adjusted at 6.8 by CO2 flushing in the solution for 15 min.
Rumen content and preparation
Rumen contents were collected from three rumen cannulated
sheep which was fed with wheat straw ad lib and commercial
concentrate mixture. The rumen contents were collected before the
morning feeding of the animals. Rumen contents were placed in pre-
warmed (39ºC) insulated flasks and transported under anaerobic
conditions to the laboratory. The rumen contents were squeezed
through four layers of cheese-cloth and kept in a water bath at 39ºC
with CO2 saturation until inoculation took place. The buffer and
inoculum (2:1 v/v) were mixed and kept in a water bath at 39ºC with
CO2 saturation (Onodera and Henderson, 1980). All laboratory
handling of rumen fluid was carried out under a continuous flow of
CO2. Buffered rumen fluid (15ml) was pipetted into each syringe,
containing the feed samples, and the syringes were immediately
placed into the water bath at 39ºC. Syringes were incubated in vitro in
water bath for 96 h and gently shaken every 2hr. The syringes were
continuing incubation up to 96 h and gas production was recorded at
3, 6, 9, 12, 24, 72 and 96 h of incubation in vitro. Total gas values
were corrected for blank incubation which contains only rumen fluid.
Cumulative gas production
The cumulative gas production (Y) at time (t) was fitted to the
exponential model of (Ørskov and McDonald, 1979).
Gas (t) = a+b×(1-exp-ct
)
Where; a = the gas production from the soluble fraction (ml), b = the
gas production from the insoluble fraction (ml), c = the gas production
rate (ml/h), and t = incubation time (h).
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J.Agric.&Env.Sci.Dam.Univ.,Egypt Vol.11 (2) 2012
Estimation of energy values, organic matter digestibility, short
chain fatty acids and microbial proteins:
The energy values were calculated from the amount of
produced gas at 24 hr of incubation with supplementary analyses of
crude protein, ash and crude fat. (Menke et al., 1979; Menke and
Steingass, 1988).
ME (MJ /Kg DM) = 1.06+ (0.157* GP at 24 h) + (0.084*CP) +
(0.22* EE) –0.08* A
OMD (%) = 14.88+0.889*gas at 24h+0.45*CP+0.0651*A
NE (Mcal/lb) = ((2.2+ (0.0272*GAS at 24h) + (0.057*CP) +
(0.149*EE))/14.64
Where: ME is the metabolizable energy, OMD is organic matter
digestibility, GP is 24 h net gas production (ml/200 mg DM), A is ash
(% of DM), NE is the net energy, and EE is ether extract or crude fat
(%of DM)
Short chain fatty acids (SCFA) were calculated according to the
Getachew et al., (2005) using the following equation: SCFA= (-
0.00425+0.0222*GP at 24h)*100
Where: GP is 24 h net gas production (ml/200 mg DM).
Microbial protein was calculated as 19.3 g microbial nitrogen per kg
OMD according to Czerkawski (1986).
2.3.4. Methane determination
Methane volume, carbon dioxide volume and the percentage of
methane in the total gas were determined according to (Fievez et al.,
2005).
2.4. Statistical Analysis
Data were subjected to analysis of variance (ANOVA) using the
General linear Model (GLM). Significant differences between
individual means were identified using least significance difference
(LSD) multiple range test (SAS, 2000).
Results and Discussion In vitro gas production
Cumulative gas production profiles, corrected for blank are
shown in table (2) and figure (1) for all treatments. The cumulative
volume of gas production increased with increasing time of
incubation. There were significant differences between the substrates
in terms of gas production at all incubation times. Supplementation
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J.Agric.&Env.Sci.Dam.Univ.,Egypt Vol.11 (2) 2012
herbs and garlic juice to the ration increased significantly gas
production (P<0.05) than those obtained by the control ration. Gas
production volume was the highest value for sample which contained
garlic juice at the level (5 ml/kg DM) (P<0.05) and was greater for
other levels of garlic juice (3, 2, 1, and 0.5 ml/kg DM, respectively)
and the ration which contained ginger at the level of (0.1%) was
higher (P<0.05) than other rations contain garlic cloves and garlic
pulp or control. Garlic juice supplementation obtained the highest
value of gas production volume and the rate of gas production at 96 h
incubation. These results agreement with Nasser (2012) and
Bunglavan, et al., (2010).
Kinetics of gas production obtained from the exponential model
is presented in Table 2. The gas production from the insoluble fraction
(b) was the highest value for ginger and garlic juice at the level of (0.1
% and 5 ml/kg DM, respectively) (P<0.05) than other treatments and
the control. Gas production rate (c) of garlic juice at the level of (5
ml/kg DM) was significantly the highest (P<0.05) than other
treatments and the control. These results are in agreement with the
results reported by Busquet et al. (2005) and (Patra et al., 2010).
Energy contents, organic matter digestibility, microbial
protein and methane emission: The predicted metabolizable energy (ME, MJ/kg DM), net
energy (NE, MJ/kg DM), organic matter digestibility (OMD%),
microbial protein (MP mg/kg DM) and short chain fatty acids
(SCFA, mM) of tested rations contained herbs are presented in Table
(3). Garlic juice supplementation obtained the highest values (P<0.05)
of SCFA than other treatments.
The present data showed that the ME and NE were higher
(P<0.05) values for both garlic juice at level (5 ml/kg DM) and ginger
with different levels than garlic cloves, garlic pulp, and control. Also,
the results showed that the OMD% and microbial protein were
higher (P<0.05) for garlic juice at the level of 5 ml/kg DM with
(67.06% and 129.42 g/kg OMD, respectively) than for others herbs
supplementation or control.
The results of In vitro total gas, carbon dioxide, methane production
and methane production (%) for 96 h incubation of the experimental
treatments showed in Table (4). It was indicated that using garlic juice
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J.Agric.&Env.Sci.Dam.Univ.,Egypt Vol.11 (2) 2012
and ginger with different levels reduced methane production with 70-
77 % compared with the control.
Also, the results showed that the highest value of carbon
dioxide production was observed with garlic juice at level (5 ml/kg
DM) than other herbs and the control. Garlic juice supplementation
and ginger with different levels obtained the lowest value (P<0.05) of
methane production volume ranged between 3.33 to 4.33 ml.
These results are agreement with Busquet et al., (2005b). They
observed, in batch culture, that garlic oil and diallyl disulphide (300
mg/l of ruminal fluid) reduced methane production by 74 and 69%,
respectively. In the experiment with sheep fed on wheat straw and
concentrate (1:1), inclusion of garlic at 10 g/kg of dry matter
decreased methane formation expressed relative to organic matter
digested (Patra et al., 2008a). Furthermore, numbers of studies have
been conducted on garlic oils or its components for modulation of
rumen fermentation and inhibition of methane production (Busquet et
al., 2005b; Chaves et al., 2008; Kongmun et al., 2010; Patra et al.,
2006c). Ohene-Adjei et al. (2008) reported that addition of garlic oil
(contained allyl mercaptan, 26%; allyl trisulphide, 18% and allicin,
1.5%) to barley-based diet (0.02 g/kg) did not affect total number of
methanogenic archea in sheep as quantified by archeal 16S rRNA
copy numbers. However, the phylogenetic analysis indicated that
garlic oil supplementation inhibited Methanogenic ruminantium. In
contrast, Methanosphaera stadtmanae and M. smithii and some
uncultured groups increased in the supplemented treatments. It has
been suggested that organosulphur compounds increased the
phylogenetic distribution of methanogenic archaea, which may have
been resulted from changes in associated protozoal species (Ohene-
Adjei et al., 2008). However, a large decrease in methane production
by garlic oils or its components in some studies (Patra et al.,
2006c; Busquet et al., 2005b) without significant changes in
protozoal populations (Patra et al., 2010) doesn't support the
hypothesis of protozoal associated methanogenesis reduction. It has
also been observed that all of the rumen ciliate protozoa may not bear
methanogens on their surface. Vogels et al., (1980) reported that
methanogens were associated with only 8% of the cells of Entodinium
longinucleatum, whereas more than half of the cells of Ostracodinium
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obtusum and Eudiplodinium maggii had methanogens on their
pellicles. No externally associated methanogens with holotrichs were
observed (Vogels et al., 1980). Even between the subspecies of one
species such differences may be present: Epidinium ecaudatum subsp.
caudatum and Epidinium ecaudatum subsp. ecaudatum showed 5 and
35% association, respectively (Vogels et al., 1980). Therefore, a
reduction of methanogenesis related to protozoa will depend upon the
species of the protozoa are affected by the phytochemicals. Because
the garlic extracts did not appreciably affect degradability of feeds
(Patra et al., 2010), it appears that garlic oil may specifically inhibit
methanogenic archaea. With the analysis by real-time polymerase
chain reaction, It been suggested that the organosulphur compounds
found in garlic oil perhaps directly inhibit the rumen methanogenic
archaea through an inhibition of the enzyme 3- hydroxy-3-methyl-
glutaryl coenzyme A (HMG-CoA) reductase (Busquet et al.,
2005b). The synthesis of the isoprenoid units in methanogenic archaea
is catalyzed by HMG-CoA reductase (Busquet et al., 2005b).
Another sulphur-containing compounds, synthetic allyl isothiocyanate
inhibited methane production and ruminal methanogenic archaea in
vitro (Lila et al., 2003b). Feeding of cyclodextrin-horseradish oil
(3.5% allyl isothiocyanate) complex to steers at 20 g/kg diet
suppressed methanogens by 20% (Mohammed et al., 2004).
In an overall conclusion it seems that, the additions of herbs
(garlic juice and ginger) improved the rumen fermentation and
reduced methane production
Table (1). Chemical composition (%) of concentrate mixture and
wheat straw and herbs.
Items DM
%
OM
%
CP
%
EE
%
CF
%
NFE
%
Ash
%
Concentrate feed
mixture
92.41 93.27 16.75 4.14 6.10 66.28 6.73
Wheat straw 91.34 89.31 2.64 1.36 38.97 46.34 10.69
ginger 93.57 91.20 7.68 6.40 5.90 71.22 8.80
garlic cloves 94.05 94.05 18.10 0.50 1.90 73.55 5.95
garlic pulp 94.69 94.69 6.25 0.30 2.01 86.13 5.31
garlic juice 5.00 99.70 3.00 0.20 0.50 96.00 0.30
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J.Agric.&Env.Sci.Dam.Univ.,Egypt Vol.11 (2) 2012
Table (2). Least square of means of cumulative gas production
volume and kinitecs parameters (ml/200 mg DM) at different
incubation times for experimental treatments.
Items
Additive
Gas production volume Kinitics parameters
a b C 12h 24h 48h 72h 96h
Control 14.3 f
21.5 d 30.5
c 32.5
c 34.2
c 0.54
bc 34.34
bc 0.043
fi
Ginger
0.1% 21.0 de
39.0 b 50.0
ab 53.0
ab 55.0
ab 0.000 58.67
a 0.04
i
0.5% 20.0 de
35.0 bc
45.0 bc
47.0 b 49.0
ab 0.000 50.97
ab 0.05
f
1% 20.0 de
37.0 bc
46.0 b 49.0
ab 52.0
ab 0.000 53.88
ab 0.05
f
2% 24.7 cd
34.7 bc
39.0 bc
40.7 bc
44.0 bc
0.000 42.28 bc
0.073 cd
3% 26.0 cd
36.0 bc
40.0 bc
40.0 bc
44.0 bc
0.000 41.95 bc
0.08 c
Garlic cloves
0.1% 19.0 e 30.0
cd 35.3
c 35.7
c 38.0
bc 0.000 38.22
bc 0.07
d
0.5% 19.3 de
30.3 c 37.0
bc 38.7
bc 41.2
bc 0.063
b 40.69
bc 0.056
ef
1% 18.7 e 31.0 c 37.0
bc 38.7
bc 41.3
bc 0.000 40.98
bc 0.056
ef
2% 15.7 ef 26.3
cd 33.7
c 35.0
c 38.0
bc 0.000 37.71
bc 0.05
f
3% 19.0 e 32.0
c 37.3
bc 38.7
bc 40.3
bc 0.000 40.92
bc 0.063
de
Garlic pulp
0.1% 18.0 e 26.7
cd 33.0
c 33.3
c 35.7
c 0.02
c 35.15
bc 0.056
ef
0.5% 17.3 ef 26.3
cd 30.0
c 30.7
c 33.3
c 0.000 33.72
bc 0.093
b
1% 18.7 e 28.7
cd 34.3
c 35.3
c 37.0
bc 0.000 36.86
bc 0.06
e
2% 17.0 ef 24.3
d 27.0
c 28.3
c 30.7
c 0.000 29.92
c 0.09
b
3% 16.0 ef 24.7
d 29.7
c 32.0
c 33.3
c 0.606
bc 32.59
c 0.053
ef
Garlic juice (ml/kg DM)
0.50 22.7 d 32.7
bc 39.5
bc 41.3
bc
42.5
bc 1.15
b 40.71
bc 0.066
de
1 25.0 cd
39.0 b 44.0
bc 45.0
bc 47.0
b 1.12
bc 45.09
b 0.07
d
2 27.0 c 39.0
b 44.0
bc 46.0
bc 48.0
ab 2.44
a 44.06
bc 0.07
d
3 32.0 b 47.0
a 52.0
ab 53.0
ab 54.0
ab 2.65
a 50.43
ab 0.09
b
5 42.0 a 52.0
a 57.0
a 58.0
a 59.0
a 0.98
bc 56.5
ab 0.12
a
LSD 3.60 6.77 10.41 10.88 11.04 1.05 12.12 0.0092
SEM 0.77 1.02 1.10 1.13 1.13 0.15 1.16 0.0008
Means followed by the same letters are not significant, but different letters are significant
according LSD procedure. LSD: Least square of means. SEM: Standard error of means.
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J.Agric.&Env.Sci.Dam.Univ.,Egypt Vol.11 (2) 2012
Table (3). Short chain fatty acids (SCFA),Metabolizable energy
(ME), Net energy (NE), Organic matter digestibility (OMD), and
Microbial protein (MP).
Items
Additive
SCFA
(mM)
ME
(MJ /Kg DM)
NE
(MJ /Kg DM)
OMD
%
MP
(g/kg DM)
Control 47.31 d 5.58
d 3.53
d 40.12
c 77.43
c
Ginger
0.1% 86.16 b 8.33
b 4.01
a 63.18
ab 121.94
ab
0.5% 77.28 bc
7.70 bc
3.90 bc
59.56 ab
114.95 ab
1% 81.72 bc
8.02 bc
3.95 bc
61.28 ab
118.26 ab
2% 76.54 bc
7.65 bc
3.88 bc
56.64 b 109.32
b
3% 79.5 bc
7.86 bc
3.91 bc
60.18 ab
116.14 ab
Garlic cloves
0.1% 66.18 cd
6.91 cd
3.76 cd
50.18 bc
96.85 bc
0.5% 66.92 cd
6.96 cd
3.77 cd
50.47 bc
97.41 bc
1% 68.4 c 7.07
cd 3.79
c 51.05
bc 98.53
bc
2% 58.04 cd
6.33 cd
3.66 cd
46.88 bc
90.49 c
3% 70.62 c 7.22
c 3.2
c 51.91
bc 100.18
bc
Garlic pulp
0.1% 58.78 d 6.39
cd 3.67
cd 44.72
c 86.33
c
0.5% 63.22 cd
6.33 cd
3.66 cd
44.43 c 85.74
c
1% 53.6 d 6.69
cd 3.72
cd 46.46
c 89.67
c
2% 53.6 d 6.00
d 3.59
d 42.55
c 82.13
c
3% 54.34 d 6.04
d 3.59
d 42.8
c 87.60
c
Garlic juice (ml/kg DM)
0.50 72.28 bc
7.34 bc
3.84 bc
50.14 bc
96.76 bc
1 86.16 b 8.32
b 4.00
b 55.67
bc 107.44
bc
2 86.16 b 8.31
b 4.00
b 55.63
bc 107.37
bc
3 103.92 a 9.55
ab 4.21
a 62.68
ab 120.97
ab
5 115.02 a 10.32
a 4.34
a 67.06
a 129.42
a
LSD 15.04 0.92 0.91 9.05 17.48
SEM 2.26 0.16 0.03 0.11 2.14
Means followed by the same letters are not significant, but different letters are
significant according LSD procedure. LSD: Least square of means.
SEM: Standard error of means.
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J.Agric.&Env.Sci.Dam.Univ.,Egypt Vol.11 (2) 2012
Table (4). In vitro total gas, carbon dioxide, methane production in ml
and methane production in percentage on 96 hour incubation of herbs
at different inclusion levels with substrate.
Additives Total gas (ml at 96 hrs) CO2 ml CH4 ml CH4 %
Control 34.16 c 19.5
d 14.66
a 42.97
a
Ginger
0.1% 55 ab
51.33 ab
3.66 d 6.66
c
0.5% 49 ab
45.00 ab
4.00 d 8.16
c
1% 52 ab
48.66 ab
3.33 d 6.41
c
2% 44 bc
39.66 bc
4.33 d 9.89
c
3% 44 bc
39.66 bc
4.33 d 9.84
c
Garlic cloves
0.1% 38 bc
29.33 cd
8.66 c 23.16
b
0.5% 41.16 bc
31.33 c 10.33
b 25.99
b
1% 41.33 bc
31.83 c 9.50 b 24.09
b
2% 38 bc
29.00 cd
9.00 bc
24.89 b
3% 40.33 bc
31.33 c 9.00
bc 23.23
b
Garlic pulp
0.1% 35.66 c 26.33
cd 9.33
bc 26.49
b
0.5% 33.33 c 24.33
cd 9.00
bc 29.83
b
1% 37 bc
28.66 cd
8.33 c 22.59
b
2% 30.66 c 22.33
cd 8.33
c 28.51
b
3% 33.33 c 24.33
cd 9.00
bc 29.96
b
Garlic juice (ml/kg DM)
0.50 42.5 bc
38.50 bc
4.00 d 9.42
c
1 47 b 43.00
b 4.00
d 8.51
c
2 48 ab
44.33 b 3.66
d 7.63
c
3 54 ab
50.33 ab
3.66 d 6.79
c
5 59 a 55.33
a 3.66
d 6.21
c
LSD 11.04 10.95 1.54 8.86
SEM 1.13 1.46 0.47 1.61
Means followed by the same letters are not significant, but different letters are
significant according LSD procedure. LSD: Least square of means.
SEM: Standard error of means.
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REFERENCES
Abdulrazak, S.A., Fujihara, T., Ondiek, J.K. and Ørskov, E. R.,
(2000). Nutritive evaluation of some Acacia tree leaves from
Kenya, Animal Feed Science and Technology, 85, 89–98.
AOAC., (1990). Association of Official Analytical Chemists. Official
Methods of Analysis. Vol. I 15th ed. AOAC, Arlington, VA.
Busquet, M., Calsamiglia, S., Ferret, A., Carro, M.D., and Kamel,
C., (2005b). Effect of garlic oil and four of its compounds or
rumen microbial fermentation. J. Dairy Sci. 88, 4393–4404.
Bunglavan, S.J., Valli, C.; Ramachandran, M.; and Balakrishnan,
V. (2010). Effect of supplementation of herbal extracts on
methanogenesis in ruminants. Livestock Research for Rural
Development, 22 (11).
Casewell, M., Friis, C., Marco, E., McMullin, P., and Phillips, I.,
(2003). The European ban on growth-promoting antibiotics
and emerging consequences for human and animal health. J.
Antimicrob. Chemother. 52, 159–161.
Chaves, A.V., He, M.L., Yang, W.Z., Hristov, A.N., McAllister,
T.A., and Benchaar, C., (2008). Effects of essential oils on
proteolytic, deaminative and methanogenic activities of mixed
ruminal bacteria. Can. J. Anim. Sci. 89, 97–104.
Czerkawski, J. W. (1986). An introduction to rumen studies.
Pergamon Press. Oxford. New York.
EC, (2003). Regulation EC No 1831/2003 of the European Parliment
and Council of 22 September 2003 on additives for use in
animal nutrition. Official J. Eur. Commun. L268, 29–43.
Fievez V, Babayemi O J and Demeyer D (2005). Estimation of
direct and indirect gas production in syringes: A tool to
estimate short chain fatty acid production that requires minimal
laboratory facilities, Animal Feed Science and Technology
123–124: 197–210.
33
J.Agric.&Env.Sci.Dam.Univ.,Egypt Vol.11 (2) 2012
Getachew, G., De Peters, E., Robinson, P. and Fadel, J. (2005). Use of in vitro rumen gas production techniques to evaluate
microbial fermentation of ruminant feeds and its impact on
fermentation products. Anim. Feed Sci. Technol. 124: 547-
559.
Greathead H (2003). Plants and plant extracts for improving animal
productivity. Proc Nutr Soc 62:279–290
IPCC, (2001). In: Houghton, J.T. et al. (Eds.), Climate Change 2001:
The Scientific Background, vol. 94. Cambridge University
Press, Cambridge, UK.
Johnson, K.A., and Johnson, D.E., (1995). Methane emissions from
cattle, Journal of Animal Science, 73, 2483–2492.
Kamra, D.N., (2005). Rumen microbial ecosystem. Curr. Sci. 89, 124–
135.
Kongmun, P., Wanapat, M., Pakdee, P., and Navanukraw, C.,
(2010). Effect of coconut oil and garlic powder on in vitro
fermentation using gas production technique.Livest. Sci. 127,
38–44.
Kiuchi, F., Shibuya, M., and Sankawa, U., (1982). Inhibitors of
prostaglandin biosynthesis from ginger. Chemical and
Pharmaceutical Bulletin (Tokyo) 30, 754–757.
Kurihara, M., Magner, T., Hunter, R. A., and McCrabb, G. J.,
(1999). Methane production and energy partition of cattle in
the tropics, British Journal of Nutrition, 81, 227–334.
Lawson, L., (1996). The composition and chemistry of garlic cloves
and processed garlic. In: Koch, H.P., Lawson, L.D. (Eds.),
Garlic. The Science and Therapeutic Application of Allium
sativum L. and Related Species. Williams and Wilkins,
Baltimore. MD, pp. 37–107.
Lila, Z.A., Mohammed, N., Kanda, S., Kamada, T., and Itabashi,
H., (2003b). Effect of acyclodextrin allyl isothiocyanate on
ruminal microbial methane production in vitro. Anim. Sci. J.
74, 321–326.
33
J.Agric.&Env.Sci.Dam.Univ.,Egypt Vol.11 (2) 2012
McGuffey, R.K., Richardson, L.F., and Wilkinson, J.I.D. (2001). Ionophore for dairy cattle: current status and future outlook. J.
Dairy Sci. 84, E194–E203.
Menke, K.H., and Steingass, H., (1988). Estimation of the energetic
feed value obtained from chemical analysis and gas production
using rumen fluid. Anim. Res. Dev. 28, 7–55.
Menke, K.H., Raab, L.; Salewski, A.; Steingass, H.; Fritz, D. and
Schneider, W. (1979). The estimation of digestibility and
metabolizable energy content of ruminant feedstuffs from the
gas production when they incubated with rumen liquor in vitro.
J. Agric. Sci. (Cambridge) 92: 217-222.
Mohammed, N., Ajisaka, N., Lila, Z.A., Hara, Koji, Mikuni, K.,
Hara, K., Kanda, S., and Itabashi, H., (2004). Effect of
Japanese horseradish oil on methane production and ruminal
fermentation in vitro and in steers. J. Anim. Sci. 82, 1839–
1846.
Nasser, M. E. A, (2012). Contribution of both soluble and insoluble
fractions of untreated and treated Acacia saligna and leucaena
leucocephala with different levels of urea to rumen
fermentation, in vitro. The International Conference of the
University of Agronomic sciences and Veterinary Medicine of
Bucharest "Agriculture for life, life for agriculture". Vol. 12,
Issue 4.
Ohene-Adjei, S., Chaves, A.V., McAllister, T.A., Benchaar, C.,
Teather, R.M., and Forster, R.J., (2008). Evidence of
increased diversity of methanogenic archaea with plant extract
supplementation. Microbial. Ecol. 56, 234–242.
Onodera, R. and Henderson, C. 1980. Growth factors of bacterial
origin for the culture of the rumen oligotrich protozoon,
Entodinium caudatum. J. Appl. Bacteriol., 48, 125-134.
Ørskov E. R. and Mc Donald, I. (1979). The estimation of protein
degradability in the rumen from incubation measurements
weighted according to rate of passage. J. Agric. Sci.
Cambridge. 92: 499-503.
34
J.Agric.&Env.Sci.Dam.Univ.,Egypt Vol.11 (2) 2012
Patra, A.K., and Saxena, J. (2010). A new perspective on the use of
plant secondary metabolites to inhibit methanogenesis in the
rumen. Phytochemistry 71:1198–1222
Patra, A.K., Kamra, D.N., and Agarwal, N., (2010). Effects of
extracts of spices on rumen methanogenesis, enzyme activities
and fermentation of feeds in vitro. J. Sci.Food Agric. 90, 511–
520.
Patra, A.K., Kamra, D.N., and Agarwal, N., (2006c). Effect of
spices on rumen fermentation, methanogenesis and protozoa
counts in in vitro gas production test. Int. Cong. Ser. 1293,
176–179.
Patra, A.K., Kamra, D.N., Bhar, R., Kumar, R., and Agarwal, N.,
(2008a). Plant secondary metabolites present in Terminalia
chebula and Allium sativum reduce methane emission in
sheep. Aust. J. Exp. Agric. 48, lxx-lxxi.
Russell, J.B., and Houlihan, A.J., (2003). Ionophore resistance of
ruminal bacteria and its potential impact on human health.
FEMS Microbiol. Rev. 27, 65–74.
SAS (2000). Statistical Analysis Systems Institute. SAS user guide:
Statistics Version 8. SAS Institute Inc., Cary. NC.
Vogels, G.D., Hoppe, W.F., and Stumm, C.K., (1980). Association
of methanogenic bacteria with rumen ciliates. Appl. Environ.
Microbiol. 40, 608–612.
Wanapat, M., Cherdthong, A., Pardee, P., and Wanapat, S.,
(2008a). Manipulation of rumen ecology by dietary
lemongrass (Cymbopogon citratus Stapf.) powder
supplementation. J. Anim. Sci. 86, 3497–3503.
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الملخص العربي
تاثير استخدام -1تأثير بعض النباتات العطرية على تخمرات الكرش:
الزنجبيل والثوم على انتاج الغاز وقيم الطاقة وهضم المادة العضوية وانتاج
غاز الميثان معمليا.
دمحم عماد عبد الوهاب ناصر* –ايمن احمد السيد نور –دمحم صالح محرم –عبدالرازق تاج الدين
جامعة دمنهور. –كلية الزراعه
جامعة االسكندرية. –* كلية الزراعة
هذه التجربة بهدف تقييم تأثير إضافة بعض النباتات العطرية أو مستخلصاتها أجريت على إنتاج الغاز وقيم الطاقة ومعامل هضم المادة العضوية والبروتين الميكروبي وإنتاج غاز الميثان
راسة طريقة إنتاج الغاز معمليا. استخدم اضافة كل من الزنجبيل والثوم حيث استخدم في هذه الد% من مخلوط العلف المركز ، بينما تم اضافة 3، 2، 0، 1.5، 1.0وتفل الثوم بمستويات صفر،
كجم مادة جافة من العلف المركز وتم /مل 5، 3، 2، 0، 1.5عصير الثوم بمعدل صفر ، ساعة فى الكرش الصناعى وكانت مادة 96، 72، 48، 24، 02، 9، 6، 3التحضين لمدد
( . استخدم فى هذه 31: 71العلف تتكون من مخلوط من العلف المركز وتبن القمح بنسبة )الدراسة ثالث كباش اغنام بلدى ذات فتحة فى الكرش لسحب محتويات الكرش وكانت تتغذى هذه
( وكانت اهم النتائج المتحصل عليها 71: 31الحيوانات على مخلوط علف مركز وتبن قمح بنسبة ) كاالتى :
ساعة 96، 24، 02استخدام عصير الثوم اعطى اعلى انتاج للغاز يليه الزنجبيل عند اوقات - من التحضين.
كجم مادة جافة اعطى اعلى انتاج للغاز عند / مل 5، 3استخدام عصير الثوم عند مستويات -جميع اوقات التحضين ، وكذلك اعطت نتائج اعلى فى كل من الطاقة الميتابولزمية والطاقة الصافية ومعامل هضم المادة العضوية والبروتين الميكروبى ،وكذلك اعلى انتاج فى االحماض الدهنية
لكربون واقل انتاج من حجم ونسبة غاز الميثان.قصيرة السلسلة واعلى انتاج لحجم ثانى اكسيد االتخمر ويزيد من انتاج من هذه الدراسة نستنتج ان استخدامات النباتات العطرية تحسن من عمليات
الطاقة وانخفاض فى حجم ونسبة غاز الميثان ،وان عصير الثوم تفوق على باقى االضافات .المستخدمة
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J.Agric.&Env.Sci.Dam.Univ.,Egypt Vol.11 (2) 2012
Figure 1: Effect of the experimental diets on gas production profiles. A= ginger, B= garlic cloves, C= garlic pulp and D= garlic juice.
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