Ruminants are unique - Ability to digest fibrous feedstuffs otherwise indigestible.

- But produce methane --a potential greenhouse gas - As an unavoidable waste product of enteric fermentation, - Global warming.
Enteric fermentation digestive process -CHO are broken down by MO simple molecules for absorption into the bloodstream.

 Globally, ruminants produce 80 MMT of methane annually.

(NRC, 2002)

India has largest livestock population in the world & estimated to emit about 10.8 MMT of CH4 annually or 405.75 x 108 Kcal/day from enteric fermentation.
(Singh and Sikka, 2007)

Dairy animals are most popular livestock enterprises in the country and account for nearly 60% of these enteric emissions.
(Singhal et al., 2005)

From agricultural sector, ruminants contribute major 49 % methane in India. (NATCOM, 2004) In ruminants, 87% CH4 is produced in the rumen & remaining 13% from hindgut fermentation.
(Moss et al., 2000) 3

GHG METHANE
Global antropogenic sources of Methane

Enteric fermentation 28% Biomass burning 5% Biofuel combustion 4%

Manure 4%

Rice 11% Natural gas 15% Coal 8%

Fuel mobile 1% Waste water 10%

Oil 1% Solid waste 13%

Source: US-EPA, 2000 http://www.epa.gov/methane/intlanalyses.html

Per cent Contribution of methane by Different Categories of Livestock (enteric fermentation and manure management).
Bovines

(Swamy and Bhattacharya, 2006)

% Emission
Cattle Indigenous Non-dairy

Buffalo Dairy Cattle Indigen- Buffalo Non- Cattle cross ous Dairy Dairy bred Dairy

Cattle crossbred Non- dairy

Small Ruminants & other Animals

% Emission

Goat

Sheep

Pigs

Camels

Horses and Ponies

Donkeys

Methane emission in India (kg/male/year)

(Singhal et al., 2005) Fig.4: Methane emission (kg/male bovine/year)
4-12 months Working
70.0 60.0
kg/male/year) /male /year) (kg

1-3 yr Breeding+ working

breeding bulls Others

50.0 40.0 30.0 20.0 10.0 0.0 Crossbred Indigenous Buffalo Average

Methane emission in India (kg/female/year)

(Singhal et al., 2005) Methane emission (kg/female/year)
4-12 months 1-3 yr Milking Dry cow s Heifers Others

80.0 70.0
kg/female/year) (kg /female /year)

60.0 50.0 40.0 30.0 20.0 10.0 0.0 Crossbred Indigenous Buffalo Average

State-wise methane emission from Indian livestock in 1994.

(Singhal et al., 2005)

Distribution of methane density (Gg/sq. km/yr)

(Swamy and Bhattacharya, 2006)

Why Inhibit Methane?

I. Loss of feed energy: 1 L of CH4 = 39.5 kJ of feed energy.

Methane emission: 8 % GEI loss. - varies with diet from 2% (cattle in feedlots) to 15% (animals eating very poor quality forage).
(Johnson and Johnson, 1995)

Disadvantage for both cows and producers as this energy could be channeled for production purposes.
II. Green House Effect & Global Warming:

Methane contributes 15 - 20% of total GHG. CH4 has 21 times more GWP than CO2. (IPCC, 1996) -Weather change (By the year 2030 the world is likely to be 12 C warmer than today). - Increase in sea level (17 - 26 cm rise in global mean sea level).

- Health hazards.

10

(Khan et al., 2001) (Moss, 1993)

 Many practices to reduce GHG emissions increase production efficiency and profitability of agricultural operations.
Producers verifiable, quantifiable GHG emission or carbon sequestration able to receive revenue from Carbon Trading markets in the future.

So, The inhibition of CH

production in the rumen would have significant economic and environmental benefits.
4

11

Methanogensis
Substrates used process include:
a) H2

in

the Need methanogens: Methanomicrobium Methanobacterium Methanosarcina

(Baker, 1998 ; Jarvis et al., 2000)

b) c) d) e) f) g) h)

CO2 Prime precursor Formate, Acetate, Methanol, Methylamines, Dimethyl sulfide, Some alcohols.

Methanogens use the process of formation of CH4 to generate energy for growth.
37% rumen methanogenesis is due to methanogens living in or on the rumen protozoa.
(Hegarty, 1999) 12

In Rumen
Dietary starch and plant cell wall
Primary fermenters
Ruminobacter amylophilus, Streptococcus bovis, R. albus, F. flavifaciens, F.succinogenes

(Mc Allister et al., 1996)

Sugars and protein Pyruvate Formate

Pyruvate formate

Secondary fermenters
Megasphaera elsdenii Treponema bryantii

VFA+CO2+H2

lyase system

VFA+CO2 +H2 +NH3

CO2 + 4H2
Methanogens, anaerobic condition More negative than -200 mV,

No electron acceptor
13

G= -31 Kcal

CH4 + 2 H2O

Importance of Methahonogenesis
H2 produced by microbes in the rumen Methanogensis Uses H2 to reduce CO2 to form CH4 If we eliminate methanogens Reduces CH4 But leaves high partial pressure of H2 Inhibition of NADH reoxidation & ruminal Fermentn Reduced digestion of fiber and microbial growth. Elimination of methanogens without alternative route of electron transfer affect the enteric fermentation.

(Wolin et al., 1997)

14

Possible intervention sites for lowering ruminant methane.

CO2 CH4

Microbes involved in digestion

H2

Methanogens

Hydrogen formation inhibitors

1. Alternative H2 sinks 1. Anti-methanogens 2. Increase microbial 2. Removal of protozoa growth yields

(Joblin, 1999)
15

Influencing Factors
Internal factors:Methanogens, protozoa, feed residence time, animal species & its production level.

External factors:
Diet composition. Level of feed intake. Forage processing. Feeding frequency. Environmental factors.
( Jarvis,2000 : Lee et al.,2003) (Johnson et al., 1993) ( Takashi, 2001 ; Santoso et al., 2003) (Balch,1960) (McAllister et al.,1996)

Internal factors are under strong influence of external factors.

16

Methane Abatement Options in Ruminant

Dietary Manipulation
Concentrate proportion Conc. type digestibility of forage Leguminous fodder Molasses/ UMNB

Manegement
Animal numbers Forage quality Animal Productivity

Rumen manipulation
Genetic engineering Defaunation

Efficiency/ Less RFI Longevity of animals

Antibiotics Bacteriocin
Vaccines Acetogenes Probiotics

Fats,Oil

Additives
Tannins, saponins

Grazing manegment

Essential oils
Propionate enhancers/Organic acids: fumarate, malate Direct Inhibitors: bromochloromethane amichloral, chloroform,chloral hydrate etc Ionophores: monensin, lasalocid, salinomycin

Methane oxidizers

17

1. Increasing Animal Productivity

(quite tough)

Increased productivity/animal dairy cows

(Kirchgesser et al., 1995)

30 25 20 15 10 5 0

25.8 21.6 18.3

g CH4/kg milk

g CH4/kg milk

4000

5000 Annual milk production

6000

Annual milk production (l) Increasing milk yield from 4000 to 5000 kg/yr increases annual CH4 emissions, but decrease emissions per kg of milk by 16% & 29% for 4000 to 6000 kg/yr for a 600 kg cow.

19

Methane emission per kg of milk in dairy cow

(Tamminga et al., 2007)

CH4 (g/kg milk)

Milk (kg/d) 30% reduction by increasing milk prod. from 10 to 25 kg/d.

20

2. Reducing Livestock Population & Longevity

(Frank OMara, 2004)

Desirable economically also, considering the feed and fodder shortage in our country. May not be practically feasible strategy until: Shift in the rural areas from subsistence to commercial dairy farming National policy on slaughtering of economically unviable animals
religious and social taboos on animal slaughtering in India.
21

Effect of Age at Slaughter on Lifetime Methane Production

200 150 100 50 0 0 4 8 12 16 20 24 28 25 month 25 month 30 month

16.5 % reduction in lifetime emissions and 12% reduction in emissions per kg carcass by going from a 30 to 25 month slaughter 22

3. Management Practices:
Pasture management:
forage species selection inclusion of legumes. continuous vs. rotational grazing strategies.
(McCaughey et al., 1999)

Management-intensive grazing:
- BMP

- more efficient utilization of grazed forage crops via controlled rotational grazing - efficient conversion of forage into meat and milk.
(DeRamus et al., 2003)

Animal selection for increased production Use of growth promoting agents Application of more refined ration balancing technologies.
(Wittenberg, 2003) 23

Monthly CH4 emission in beef cows on Best Management Practices v/s Conventional Forage Management Systems
(De Ramus et al., 2003)

Annual CH4 emissions in cows reflect a 22% reduction from BMP when compared with continuous grazing. 24

4. Effect of Residual Feed Intake

Low RFI group have lower MPR and reduced methane cost of growth (24% of total CH4/d & 25% of CH4/kg ADG).
(Hegarty et al., 2007)

MOA: Differences in metabolizability Genetic link between methanogens and their hosts.
(Hackstein et al., 1996)

Useful for the efficient planning of breeding strategies to select animals that eat considerably less to achieve a similar growth rate and body wt.

25

Relationship of feedlot RFI with methane production

(Nkrumah et al., 2006)

Trait High
Methane, L/kg of BW0.75
Intake energy, kcal/kg of BW0.75 Methane energy, kcal/kg BW0.75

RFI group Medium

1.68 c
382.24 15.90cd

Low
1.28d
387.98 12.09d

1.71c
384.77 16.08c

Methane Energy loss % of GEI

4.28c

4.25c

3.19d
(25%)

28% & 24% less total CH4 prod. in low- RFI than high & medium RFI animals.

26

Dietary manipulation
(strategic feeding)

Principles of reduction in Enteric CH4 Production

Reducing H2 & CO2 production (Ionophores). Reducing protozoa or Defaunation- prevention of interspecies H2 transfer ( Fats, Saponins, UMMB). Enhancing propionate production (Acrylate, Organic acids) Rechanneling H2 & CO2 to acetate (Enhancing acetogenesis). Reducing or killing methanogens ( AQ, BES, Vaccines). Use of alternate electron sink ( OA, Nitrate, Sulfate). Enhancing hydrogen utilizers (Microbes) .
28

5. Effect of Concentrate

A. Effect of Concentrate proportion

Negatively correlated with methane emissions.
(Holter and Young, 1992; Kurihara et al., 1998; Yan et al., 2000)

propionate acetate (and sometimes butyrate). rumen pH Methanogens are pH sensitive animal performance
(Johnson and Johnson, 1995)

Forage replaced by concentrate rich diet lowered CH4 production by 40% (from 272 to 170 g/day).

Limitations: Required minimum level of physical structure in the diet Balance between energy intake and requirements

(Veen. 2000)

(Kadokawa & Martin, 2006)

30

The effect of increasing the Proportion of Concentrates in the diet

(Yan et al., 2000)

methane output

Proportion of Concentrate
Total DE,DMI and feeding level are kept constant
31

Effect of Concentrate Level on Methane Emissions and their relationship to animal productivity.
(Lovett et al., 2005)

Item
CH4 (g/d) CH4 (g/kg of DMI) CH4 (g/kg of milk) CH4 (g/kg of FCM) CH4 (g/kg of milk protein) CH4 (g/kg of milk fat)

Low concentrate
(0.87 kg on DM basis)

High concentrate
(5.24 kg on DM basis)

346 19.60 21.0 19.26 555 525

399 17.83 (9%) 17.7 (15%) 16.02 509 428

Free - ranging dairy cows Effective within dairy industry

32

Methane production by lactating buffaloes under different feeding regimes

Parameters Total DM intake (kg)
Milk yield(kg/d/animal)
Berseem + Wheat straw 14.68
Berseem+ wheat straw + Concentrate Wheat straw + Concentrate

9.90

9.28

7.01

7.42

7.61

Methane production
Total production (g/d)
259.74 a 17.76 a 22.31 b 162.67 b 16.52 ac 22.08 b 177.03 b 19.13 ab 30.20 a

g /kg DMI g /kg DDMI

g /kg milk yield

38.16 a ( Singhal et al., 2006 )

22.12 b

23.45 b
33

B. Effect of Concentrate Type

Variable to: Structural (cellulose, hemicellulose) Non-structural (starch, sugars) carbohydrates.
(Ovenell-Roy et al., 1998)

CH4 emision: For every g of cellulose digested = 3 hemicellulose = 5 soluble residue. (Moe and Tyrrell, 1979) Soluble sugars > potential than starch.
(Johnson & Johnson, 1995)

Cattle in finishing phase fed with barley - 2.8 % corn based diets - 4.0% of GEI CH4 losses accounted.
(Beauchemin & McGinn, 2005)

Feeding more concentrates per cow, with a higher amount of (rumen resistant) starch and less sugars has a very positive effect.
34

6. Effect of Forage Quality

Digestibility of cell walls in forages lower CH4 emission.
(Johnson and Johnson, 1995)

Forage maturity and physical form influence CH4 production: Higher for: mature forage vs. immature forage, coarse chopped vs. finely ground or pelleted low quality forage,

hay vs. silage.

35

Effect of Forage OM Digestibility on CH4 Emission

(Boadi et al., 2002)

Parameter High

Forage Quality
Medium
50.7

Low
38.5

IVOMD, %
DMI, kg/d CH4, L/d

61.5

Ad-libitum
9.7a 281.7a 8.9a 289.8a 6.3c 203.5b

CH4, %GEI
DMI, kg/d CH4, L/d

6.0

7.1

6.9

Restricted Intake (2% BW)

6.4 224.6 6.1 193.3 6.1 195.6

CH4, %GEI

7.6

7.1

7.1
36

7. Effect of Forage Type

Forage legumes: (lucerne or red clover)
less CH4 emisssion (g/kg DMI) than grass.
(Ramirez and Barry, 2005)

Due to: - lower proportion of SC and faster passage rate. - Presence of tannin, saponin.
(McCaughey et al., 1999)

Fresh grass < Grass silage (CH4 emission)

Maize silage and Whole Cereal Plant silage (WPS):
High starch, less CH4 emission.
(Tamminga et al., 2007)

Replacing half of the 60% of grass silage with maize silage in midlactation dairy cattle reduced CH4 production from 6.0 to 5.8% of GEI.
(Van Laar and Van Straalen, 2004)
37

8. Use of Molasses/ UMB

Cost effective Potential to reduce CH4 emissions by 25 to 27%.
(Bowman et al., 1992; Robertson et al., 1994)

In India CH4 reduction from 10 to 15%.

(Singh 2001; Srivastava et al., 2002)

Increase milk production at the same time. Chemical upgrading of poor quality roughage.
38

9. Effect of Dietary Oil/ Lipids

Bio-hydrogenation of unsaturated fatty acid. Enhances propionic acid production. Protozoal inhibition.
(Dohme et al., 1997)

Coconut oil addition at 3.5 and 7.0% reduces CH4 production by 28 and 73% respectively. (Machmullar and Kreuzer, 1997) Addition of mustard oil (68 mg) in WS + Conc. based diet reduced 18% of methane production in vitro.
(Tyagi and Singhal, 1998)

Fish oil at 7.5% reduces 80% methane production.

Disadvantage:
High oil prices and sustainability

(Fievez et al., 2003)

Decreases DMI and Fiber digestion (at 5% inclusion level).

(Barun Bairagi and Mohini, 2005 )
39

Effect of Refined Soy Oil or Whole Soybean

(Jordan et al., 2006)

Item

Treatment

Control
DMI, kg CH4, L/d CH4, L/kg of DMI CH4, % GEI

Whole soybean Refined soy oil

(6% of DMI) (6% of DMI)

7.88
137.8 17.9 3.9

6.32
103.0 (25%) 15.2 3.7 (5%)

7.52
83.9 (40%) 11.2 2.3 (41%)

CH4, L/kg of ADG

CH4, L/kg of ADCG Protozoa, 106/mL

99.4
141.4 1.87

89.3
125.1 1.19

56.0
85.2 0.88
40

Control: Barley/Soybean meal-based concentrate, (F:C = 10:90)

High-forage diet with Canola Oil - CH4 emissions

(Beauchemin and McGinn, 2006)

Item Control DMI Methane g/heifer g/kg of DMI % of GE intake % of DE intake

6.38a 159.3a 25.5 7.93ab 12.76

Treatment Canola oil

5.07b 108.0b (32%) 21.7 6.30a (21%) 11.97

Angus heifers fed Canola oil (6% in DM) + barley silage (75%) + 19% barley grain Reduced CH4 emissions by 32%. 41

A high-forage diet with Additives

(McGinn et al., 2004)

Item

Treatment

Control
DMI, kg/d Methane g/steer g/kg of DMI 7.40 166.2b 22.64b

Sunflower oil (1mL/kg DM) (33mg/kg DM) (5% of DMI) 7.55 7.71 6.91
164.4b 22.11b 159.6b 20.70b,e 129.0c (22%) 18.81c

Enzyme

Monensin

% GE intake
% DE intake

6.47b
10.51bc

6.32b (3%)
11.27b

5.91b,e (9%)
9.31cd,e

5.08c (21%)
8.76d
42

In beef cattle, the addition of sunflower oil (400 g/d or 5% of DMI) decreased CH4 emissions by 22% without -ve effect on DMI.

Effect of crushed seed oil on in vitro methanogenesis

(Machmuller et al., 1998)

Without Rumen Protected fat fat

Coconut oil

Rape- Sun Linseed seed flower seed

43

10. Plant Secondary Metabolites

Potential plants / tropical MPT

bioactive secondary plant metabolites (saponins and tannins) able to suppress methanogenesis.
(Carulla et al., 2005; Hu et al., 2006)

A. Condensed tannins:
Mechanism: 1) Indirectly reduction in fiber digestion
2) Directly inhibit the growth of methanogen.
(Tavendale et al., 2005)

H2 prod.

Cichory (Chichorium intybus) promising forage also reduce CH4 emission. (Ramirez & Barry, 2005)
Risk of bloat
(Singh and Sikka, 2007)
44

B. Saponin
No direct effect but reduce protozoa (50-60%). 70% methanogenes associated with protozoa.
(1983,Moss et al., 2001)

Tea saponin (0.2 and 0.4 mg/ml) decreased (12.7% and 14.0%) CH4 emission in in vitro.
(Hu et al., 2005)

Source:
Extract of soapnut Yucca extracts Fruit of Sapindus saponaria Leaves of Entrobium timoba Methanol extracts of Sapindus rarak fruit Tree leaves of Enterolobium ciclocarpum
(Kamra, 1993) (Wallace et al., 1994) (Diaz et al., 1999) (Gupta et al., 1993) (Thalib et al.,1996) (Alberto et al., 1992)
45

The effect of Condensed Tannin, PEG, BES

(Tavendale et al., 2005)

Parameter

Medicago sativa
Control PEG BES 6.47

Lotus pedunculatus
PEG + Control BES 6.21 6.14 PEG 6.50 BES 6.30 PEG + BES 6.26

pH

6.48

6.48

CH4(ml)
CH4 (%)

12.51
18.13 0.14 0.20

11.88 1.80
11.88 1.80 0.17 0.24 6.55 10.13

1.63
1.63 6.57 10.53

8.80
15.19 0.07 0.11

10.58 1.86
16.25 3.86 0.14 0.22 1.94 3.84

1.95
3.31 3.30 5.73

H2(ml)
H2 (%)b

Medicago sativa: - condensed tannin, Lotus pedunculatus: + condensed tannin Measured after 12 h.

PEG (to bind condensed tannins) BES (to inhibit methanogens)

46

11. Direct Inhibitors

Halogenated methane analogues and related compounds.
(Van Nevel and Demeyer, 1995)

Chloroform

(Bauchop,1967)

Chloral hydrate- converted to chloroform in rumen.

(Prins, 1965)

Amichloral- (a hemiacetal of chloral and starch): safer but activity declined with prolonged feeding.
(Johnson, 1974)

Trichloroacetamide and trichloroethyl adipate

(Clapperton, 1974 & 77)
47

Cont.

Bromochloromethane

(Sawyer et al., 1974)

Combination of bromochloromethane and -cyclodextrin

(May et al., 1995)

2-bromoethanesulfonic acid (BES):

Specific potent inhibitor of methanogen, not inhibit other bacteria.
(Sparling and Daniels, 1987)

9,10-anthraquinone:
Naturally occurring glycoside.

Methyl Co A

Methyl reductase Demethylation

CH4
(Odom et al., 1995)
48

Inhibition by Halogenated Compounds

Comp. TCE1 Amount /day 120 mg 240 mg Inhibition (%) 21-53 53-59 Animal Sheep

HCS2 TCA3
CH4

2.2 g 120 mg
4g

50-82 94
100

Sheep Sheep
Sheep

BCM5
BES6

5.5 g
0.03 mM

100
76

Steers
In vitro

1) Trichloroethyl adipate 2) Hemiacetyl of chloral & starch 3) Trichloroacetamide 4) Chloral hydrate 5) Bromochloromethane 6) Bromoethane sulfonic acid
49

Effect of 2-BES on methane emission from Lucerne (L) & Maize (M)
(Agarwal et al., 2005)

40

Methane ml/gDM

30
L L+BES M M+BES

20

10

0 4 8 12 24
50

Effect of 9,10-AQ (10 ppm/12 h)

(Garcia-Lopez et al., 1996)

In vitro microbial continuous culture fermentation 10:90 forage concentrate diet.

51

12. Use of Ionophores

Monensin, Rumensin, Lasalosid, Salinomysin:

- shift the H2 producing gram +ve

gram ve

- shift in fermentation from acetate to propionate - Reduction in voluntary feed intake causing fermentation.
(Goodrich et al., 1984)

- Selectively acetate (and therefore H2) production.

(Slyter, 1979)

- Inhibiting the release of H2 from formate.

(Van Nevel and Demeyer 1977)

Rumen microbes adapt to monensin on continuous feeding.

(Johnson & Johnson, 1995; 2002)
52

Effects of Monensin lactating dairy cows

(Odongo et al., 2007)

Response*

TMR

Control
DMI, kg/d
19.7

Monensin
19.1

Methane production, g/d

Methane production, g/kg BW

458.7a
0.738a

428.7b (7%)
0.675b (9%)

Methane production, g/g NDFI

0.069

0.066

(p<0.05) TMR (Forage : concentrate=60:40) Monensin 24 mg Rumensin Premix/ kg DM).

53

Ionophore feeding on Ruminal Fermentation

(Guan et al., 2006)

* = P<0.05
Low-concentrate diet - ionophore (C); + monensin (M); or + either monensin or lasalocid (M/L) which were rotated every 2 wk in yearling steers.

54

Effect of Ionophore and Dietary Cation (2.5%)

(Rumpler et al., 1986)

lonophore group

Days after beginning on ionophore 2-3

113a 96a 100a 103a

Cation treatment Control Na K

12-13
107a 123b 115a 115b

22-23
124a 124b 117a 120b

CH4 production, l/d

CH4 production, l/d 107a 123b 114ab 115ab 109a 100a 101a 103a 124a 110ab 124b 119b

No ionophore Monensin Lasalocid Avg

Steers fed a high grain basal diet Na addition CH4 prod. in the monensin group by 19% No significant effect of K addition on CH4 prod.

P< 0.05

55

13. Propionate enhancers or Alternative hydrogen acceptors:

Dicarboxylic organic acids:
Malate, Fumarate, Citrate, Succinate, Acrylate: i) Alter rumen fermentation as ionophores. (Martin, 1998) ii) Precursor of propionate (Ouda et al, 1999)

Reduces CH4 production - In vitro (Asanuma et al., 1999; Newbold et al., 2002) - In vivo (Newbold et al., 2002) - The response was dose dependent.
(Martin and Streeter, 1995)
56

Effects of Myristic Acid - Lactating Dairy Cows

(Odongo et al., 2007)

Response DMI, kg/d

Control (TMR) 15.2

Myristic acid (5% on DM basis) 14.2

Milk yield, kg/d

Milk fat, % Milk protein, % Methane production, L/d Methane production, MJ/d

14.9
4.2 3.5 608.2 23.0

13.4
4.1 3.6 390.6 (36%) 14.8 (36%)
57

Effect of Malate in vitro

(Carro and Rannilla, 2003)

Parameter

Malate (mM)

0
(0 mg/g)

4
(53.6 mg/g)
6.13 1.11 3.94 1.43

7
(93.8 mg/g)
6.16 1.10 4.12 1.42

10
(134 mg/g)
6.21 1.10 4.06 1.38

pH CH4 (mM) TVFA (mM) A:P

6.12 1.20 3.72 1.50

RL from merino sheep fed lucerne hay + concentrate (300g) Malate source: Rumelato ( Disodium malate+ calcium malate 16:84)

58

Effect of Fumarate in vitro

(Asanuma et al., 1999)

Fumarate (mM) Added 0 20 30 Consumed 0 16.0 27.0

CH4 ( mM/L culture) 15.2 14.4 13.5

VFA produced (m M)

Final pH

A
29.3 33.0 32.6

P
12.7 19.8 20.0

B
6.7 6.1 6.6 6.3 6.4 6.6

Substrate: hay powder and concentrate RL from goat 20h post feeding
Here 20 & 30 mM fumarate 34.8 & 52.2 mg/g substrate
59

14. Essential Oils

Plant extracts

Terpenoids and Phenolic compounds,(e.g monoterpenes, limonene, thymol, carvacrol) have antimicrobial activity.
EuO and black seed extract could be a promising methane mitigating agent in vitro.
(Sallam et al., 2007) The specific mode of action poorly characterized or understood.

60

Effect of plant extracts in vitro

220 200 180 160 140
GP

Thyme

65

Fennel Ginger B.seed

Thyme Fennel

60 55 50 45 40
CH4

Ginger B.seed

120 100 80 60 40 20 0
Co ntr ol T0 .5 T1 .0 T1 .5 F0 .5 F1 .0 F1 .5 G0 .5 G1 .0 G1 .5 BS 0. 5 BS 1. 0 BS 1. 5

35 30 25 20 15 10 5 0
Control F0.5 F1.0 F1.5

BS0.5

BS1.0

Plant Extracts

Effect of different levels of plant extracts on gas production et al., 2007 (GP, ml/g DM) in vitro in cubation forSallam 24 hr.

Plant extracts Effect of different levels of plant extracts on methane production (ml/g DOM) in vitrofor 24 hr incubation

61

BS1.5

T0.5

T1.0

T1.5

G0.5

G1.0

G1.5

Effect of different levels of Eucalyptus Oil - in vitro

140 120 100 GP (ml/g DM)

35 30 25

80

20
60 40 20 0 Control 25l 50l 100l 150l

CH4

15 10 5

EuO levels

0 Control 25 l 50 l EuO levels 100 l 150 l

Fig.2 Effe c t of diffe r e nt le ve ls of EuO on me tha ne pr oduc tion (ml/g DOM) in vitr o

Sallam et al., 2007

62

Result of Dietary Manipulation

(Singh and Sikka, 2007) Method Extent of Reduction (%)

Increase in Concentrate mixture Supplementation of Monensin a) Maintenance diet

20 32

14 23

b) Medium production diet

c) High producing diet Supplementation of UMMB Supplementation of green fodder

23 32
14 25 10 11 11 27
63

Rumen Manipulation

15.Stimulation of Acetogens:
Produce acetic acid by the reduction of CO2 with H2.

As daily feed additive CH4 prod. in vitro.

(Lopez et al., 1999)

Do not compete in rumen with methanogens. (10 to 100 times higher affinity for H2)
(Joblin, 1999)

Genetically modified acetogens can compete more effectively in the rumen.

65

16. Methane Oxidisers:

Oxidation of methane to CO2 Methane oxidising bacteria isolated from the rumen.
(Hanson, 1992)

Methane oxidising bacterium from the gut of young pigs when added to rumen fluid in vitro CH4 accumulation.
(Valdes et al., 1997)

66

17. Defaunation:
Removal of protozoa from rumen. Reduces CH4 production by 20 - 50 % & Improves the feed utilization efficiency. (Pal et al., 1994)

Defaunating agents:
Plant secondary metabolites (Saponin)- without inhibiting bacterial activity. (Kamra, 2005; Thalib et al, 1995) Fat (USFA)
(Mudgal et al., 2003)

USFA inhibit methanogenesis even in the absence of rumen protozoa.

Toxicity of long chain FA to methanogen.

(Dohme et al, 1999)

Pelleted concentrates fed.

(Kreuzer and Kirchgessner, 1987)

67

18. Probiotics
Aspergillus oryzae (AO) - 50% CH4 prod. directly reducing protozoal population (45%).
(Frumholtz et al., 1989)

Saccharomyces cerevisiae (SC) - CH4 by 10% (in vitro).

(Mutsvangwa et al., 1992)

Mode of action of probiotics: Reduce H2 availability to methanogens. Increased butyrate or propionate (Martin et al., 1989) Reduced protozoal numbers (Newbold et al., 1998) Promotion of acetogenesis as a sink for hydrogen.
(Chaucheyras et al., 1995)
68

19. Immunisation
Immunise ruminants against their own methanogens.
(Baker,1995)

Developed vaccine containing methanogenic microorganisms.

Ag

derived

from

rumen

(Baker, 1998)

Vaccine: cost-effective and long-acting to reduce CH4 emission and enhance animal production under grazing. Natural development of Ab by lambs against mixed methanogens.
(Holloway and Baker, 2000)

In small ruminants (sheep) reduces CH4 emission by 7%.

(Baker,1995)

Development of a vaccination strategy is in progress.

69

20. Bacteriocins
Antibiotics- protein or peptide in nature, produced by bacteria. Many lactic-acid bacteria produce bacteriocins- effect on methanogens rather than a direct pH effect.
(Russell, 1998)

Some strains of Butyrivibrio produced inhibitory activity.

(Kalmakoff et al., 1996)

Lactococcus lactis (Nisin) - stimulate propionate production & 36% CH4 (in vitro).
(Callaway et al., 1997)

Further research required to establish their adaptability & long term effectiveness as a feed additive for methane supressor.
70

Evaluation of Mitigation Strategies in Indian Context

(Sirohi et al., 2007)

Applicable to all types of dairy and non-dairy animals who are on poor diet. Cost of the technology for indigenous cows and buffaloes.

I. Strategic Supplementation Using Molasses-Urea Products (MUP) Assumptions:

(a) Adult animal 400 g/day & heifers 300 g/day. (b) CH4 reduction 11% /animal. (c) Milk yield increased by 10%. Existing avg lactation yield: local cows 1.8 litres/day (270 d lactation length) buffaloes 4.0 litres/day (280 d). (d) The cost of MUP 0.089/ kg. (e) Selling price of cow milk - 0.14/litre buffalo milk - 0.16/litre.

71

Economics of MUP supplementation

Type of animals Annual cost of supplem entation (C) Annual CH4 emission Annual CH4 reduction (RE)@11 % Gross cost of reduction Gross cost of reduction Increased return from milk production (RM)@10% Net cost of reduction Net cost of reduction

Units

Kg/head

Kg/head

/kgCH4

/t CO2

/kgCH4

/t CO2

Dairy Animals Adult animal:

Local Cow
Buffalo

13.0
13.0 9.7 9.7

36
77 22 37

3.96
8.47 2.42 4.07

3.3
1.5 4 2.4

156.3
73.1 191.8 114.0

6.9
18.0 -

1.5
-0.6
-

72.7
-28.1

Heifer:
Local cow Buffalo

Non-Dairy Animals
Adult male: Cattle Buffalo 13.0 13.0 34 55 3.74 6.05 3.5 2.1 165.4 102.3 72

II. Economics of Increased Concentrate Feeding

Assumptions: Less than 500 g conc. /animal/ day. or 7.5% conc. (not sufficient) Recommended nutritional requirement for the Indian cattle: Concentrate: Roughage = 40 : 60 (DM basis). C :R = 50:50 (for high milk producing dairy animals)
Type of animals Annual cost of addi. Conc. (C) Annual CH4 emission Annual CH4 reduction (RE) @11% Gross cost of reduction Gross cost of reduction Increased return from milk production (RM)@10% Net cost of reduction Net cost of reduction

Units

40.2 80.3 100.4 80.3

Kg/h 36 77 39 35

Kg/head 5.4 11.6 5.9 5.3

/kg CH4 7.4 6.9 17.0 15.2

/t CO2 354.1 329.6 810.1 721.5

6.9 72.0 141.6 -

/kgCH4

/t CO2 292.8 34.1 -322.8 73

Dairy Animals
Local cow Buffalo Crossbred cows

6.1 0.7 -7.0

-

Non-Dairy Animals

III. Feed Additive Ionophore

(Monensin Sodium Salt)

Assumptions:
(a) 100 mg of monensin/ animal/d.
(b) CH4 reduction potential: maintenance ration : 20%, (Indigenous animals) medium production ration: 30%, (buffaloes) high production ration : 22%, (crossbred) (c) Increase in milk yield is 5%. (d) Rumensin ( 200g monensin -Na salt/ kg) costs 7.4/ kg. Pure monensin sodium salt costs about 35,714/kg.
74

Cost through Ionophore Feed Additive

Type of animals Annual cost of ionopho re (C) Annual CH4 emission Annual CH4 reduction (RE)@11% Gross cost of reduction Gross cost of reduction Increased return from milk production (RM)@10% Net cost of reduction Net cost of reduction

Units

Kg/head

Kg/head

/kg CH4

/t CO2

/kg CH4

/t CO2

Monensin premix (Rumensin)

Local Cow
Buffalo Crossbred cows Other animals

0.27
0.27 0.27 0.27

36
77 39 35

7.2
23.1 8.6 7.0

0.04
0.01 0.03 0.04

1.8
0.6 1.5 1.8

3.5
9.0 14.2

0.4
0.4 1.6

21.2
-18.0 76.9

Pure Monensin sodium salt

Local Cow 651.78 36 7.2 90.5 4310.7 3.5

90.0

4287.8

Buffalo
Crossbred cows Other animals

651.78
651.78 651.78

77
39 35

23.1
8.6 7.0

28.2
75.8 93.1

1343.6
3609.0 4433.9

9.0
14.2

27.8
74.1

1325.0
3530.6
75

Conclusions
1. Many recommended on-farm practices will reduce
enteric CH4 and total GHG emissions by reducing feed costs associated with animal maintenance: Increasing productivity per cow to reduce methane emissions per kg of milk. When total output levels (e.g. total milk or beef produced) remain constant and livestock numbers are reduced. A lower culling age Best management practices and pasture management. Selection of Low RFI animals that achieve similar growth rate and body weight.

2. By dietary manipulation through:

Increasing proportion of concentrate rich in starch, High quality leguminous forage, Grinding and pelleting of forages, Inclusion fats and oils, TMR and UMMB, Organic acids (fumarate, malate) Tannin and saponin rich plants, Ionophores, like monensin, lasalocid, silnomycin Direct inhibition of methanogensis by using halogenated methane analogues (BES, AQ) is not found suitable.

3. Through Indirect ways of altering rumen ecology :

- Immunisation, - Elimination of ciliate protozoa, - Enhanced bacteriocin production, - Enhanced acetogenesis - Probiotics or Yeast cultures

4. Biotechnology can play an important role by manipulating the rumen bugs to enhance the digestibility of poor quality feed stuff and lower or halt the production of methane. 5. Indian senario:
Under Indian field conditions 25% CH4 reduction can be achieved. Due to poor genetic potential and low lactation yield, the net cost of CH4 reduction is very high in local cows.
The gross cost will range from Rs. 39.2 -100.8/kg CH4.

CH4 mitigation in local cows and buffaloes through increasing concentrates in the diet is a more expensive option.

Use of monensin premix is encouraging and the most cost effective technology.
Strategic supplementation (energy, nitrogen and minerals) using UMP is the second less expensive and feasible abatement option.

Thank You

Methane