EFFECT OF SOME HERBS ON THE RUMEN FERMENTATION: 1- … · Ginger (Zingiber officinale) has been...

33

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


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.

33


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


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.

33


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).

33


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).

33


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

34


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

34


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

34


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

33


33


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.

33


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


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.

33


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


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.

33


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.

http://www.lrrd.org/lrrd22/11/cont2211.htm

http://www.lrrd.org/lrrd22/11/cont2211.htm

33


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


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


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.

34


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

34


Figure 1: Effect of the experimental diets on gas production profiles. A= ginger, B= garlic cloves, C= garlic pulp and D= garlic juice.

33


EFFECT OF SOME HERBS ON THE RUMEN FERMENTATION: 1- … · Ginger (Zingiber officinale) has been...

Documents

Transcript of EFFECT OF SOME HERBS ON THE RUMEN FERMENTATION: 1- … · Ginger (Zingiber officinale) has been...