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AN IN-VITRO STUDY OF SOME SPICES FOR REDUCING METHANOGENESIS IN
RUMINANTS
ABSTRACT
The objective
of this study was to systematically evaluate and compare the effect of selected
spices on methanogenesis using in vitro cultures. Some of the spices selected
are Allium cepa, piper nigrum. Aframonum melegueta, Dennttia tripetala,,
syzygium aromaticum, Gongronema latifolium among others were dried and milled
for chemical analysis and in vitro gas fermentation study. The results showed
that the organic matter was highest in Allium crispum (97.98%) and lowest in
Allium cepa (96.77%) ash content ranges from 4.2% to 2.34% for Allium cepa and
Allium crispum respectively. The crude protein (CP) also was found to vary from
24.50% in Allium cepa to 11.19% for Gongroneme latifolium. NDF values ranges
from 52.00% for Capsicum annuum to 14.43% for Allium ascalonium and ADF values
was 40.50% for Capsicun annuum to 4.25% for Allium crispum, also hemicelluloses
was noticed to be highest in Allium crispum (39.00%) and lowest in Allium
cepa(white) (2.75%). The effect of spices was also observed on the dry matter
digestibility (DMD), methane gas production and reduction and short chain fatty
acid production. For digestibility shea butter gave the highest value of 96.15%
while the lowest was noticed in Vernonia amygdalina (61.54%). Dennttia
tripetala which had the least % methane production (25.02%) also had the
highest percentage of methane reduction (47.69%) and short chain fatty acid
(0.66mmol). This reduction in methane gas could be attributed to the bioactive
substances such as tannin(0.09%) and saponin (0.20%) present in the spices.
These results reveal that spices could be used to enhance digestibility,
increases the energy value of feed and reduces methanogenesis which is of great
concern worldwide. Spices such as Aframonum melegueta, Allium cepa, Vernonia amygdalina,
Allium crispum, Cymbopogon citratus, Dennttia tripetala, Allium cepa(white),
Syzygium aromaticum, Gongronema latifolium, Capiscum annuum that have the same
degree of methane reduction potential like Dennttia tripetala are recommended
for this purpose
TABLES OF
CONTENTS
Title Page –
——— i
Abstract – –
– – – – – – – – – – – ii
Acknowledgement
– – – – – – – – iii
Certification
– – – – – – — v
Dedication
———-vi
Table of
contents ———- vii
List of
tables ———-ix
List of
Plates———–xi
CHAPTER ONE
1.0Introduction———-1
1.2Thesis
Objectives——- –2
CHAPTER TWO
2.0Literature
Review———3
2.1What is
Tannin ———3
2.2Properties
of Tannin ——–3
2.3 Tannin
And Rumen Fermentation——-4
2.4Saponin———-5
2.5Saponin
And Rumen Fermentation——-6
2.6Spices
———-7
2.7In Vitro
Gas Production Techniques ——10
2.7.1History
of In Vitro Gas Production Techniques—–10
2.7.2Effect
Of Venting Gas During The Incubation —–11
2.7.3Effect
of Agitation of the Medium ——-12
2.7.4Effect
of Inoculum ———12
2.7.5Use Of
Blanks———13
2.7.6Effect
of Medium Composition——-13
2.7.7Effect
of Apparatus———14
CHAPTER THREE
3.0Materials
And Methods ——–15
3.1Experimental
Materials and Its Preparation Collection of Spices —15
3.2
Preparation for In Vitro study——-21
3.2.1
Preparation of Buffer——–21
3.2.2
Preparation of Inoculums ——–22
3.3Determination
of Methane Gas ——-24
3.4 Cell Wall
Fraction Determination (NDF & ADF)—–25
3.4.1 Crude
Protein Determination——-26
3.4.2 Ash
determination ———27
3.4.3 Tannin
Content Determination ——-27
3.4.4 Saponin
Content Determination ——-28
3.4.5
Moisture Content Determination ——-29
3.5Short
chain fatty acids———30
3.6Experimental
Design———30
CHAPTER FOUR
4.0Results———-31
4.1In Vitro
Gas Production at Different Hours Of Incubation Treated With
Different
Spices———31
4.2 Effect of
Spices on Dry Matter Disappearance, Absolute Methane And Short
Chain Fatty
Acid Production ——-34
4.3Chemical
Composition of some of the spices used in the study Data of the
Chemical
composition of the ten spices selected by grading are several in 4.3.36
4.4 Tannin
and Saponin Content of Some of the Spices—–39
CHATER FIVE
5.0
Discussion———-40
5.1In-Vitro
Gas Production At Different Hours Of Incubation Treated
With
Different Spices——–40
5.2Effect of
spices on dry matter disappearance methane and short chain
fatty acid
production——–40
5. 3Chemical
Composition of Some of the Spices Used in the Study —41
5.4Tannin and
Saponin Content of Some Of The Spices Used —-42
CHAPTER SIX
6.0Conclusion
And Recommendation——-43
REFERENCES———-44
LIST OF
TABLES
Table 2.1:
List of spices and there active ingredients as well as medicinal uses -9
Table 3.1:
While their pictures are shown in plate 3.1. —–15
Table 4.1:
Effect of Spices on the Volume of Gas (ml/130mg) Produced at Different
Incubation
Hours ——-33
Table 4.2:
Effect Of Species On Dry Matter Disappearance, Methane And
Shortchain
Fatty Acid Product ——-35
Table 4.3
Chemical Composition of Some of the Spices Used In the Study (%) –37
Table 4.4
Tannin and Saponin Content of Some of the Spices (%)—-38
LIST OF
PLATES
Plate 2.1:
Tannin powder———4
Plate 3. 2:
Collection of rumen fluid from a goat with a suction tube into a
Thermoflask———23
Plate 3.3:
Filtration of rumen liquor with cheese cloth —-23
Plate 3.4:
Measuring of 20ml of buffered solution into syringes —-24
Plate 3.5:
Syringes containing buffered rumen fluid being kept in an incubator at 39 o C-24
CHAPTER ONE
1.0 INTRODUCTION
Livestock is
one of the longest sources of methane emission with 80 – 115 million tons
produced per year, equivalent to 15 – 20% of total anthropogenic methane (IPCC,
2001).
Ruminants are
major contributors to biogenic methane formation. It has been estimated that
preventing methane formation from ruminants would stabilized atmospheric
methane concentrations and improve animal performance (Johnson and Johnson,
1995). The global cattle population is responsible for 73% of methane emissions
of all livestock and methane produced during ruminal fermentation represents a
loss of 2-15% of gross energy intake and may also known as a great contributor
to global warming which is a primary environmental concern world wide (moss et al.,
2000).
Recent
studies have shown that plant secondary metabolites such as tannin, essential
oils and Saponin at lower concentration could be used to manipulate rumen
fermentation favorably.
Plant
bioactive (PB) or plant secondary compounds are chemicals synthesized in plants
but are not involves in the primary biochemical processes of plant growth but
acts as a protective agents against predators.
Plant
extracts from spices and medicinal plants with high concentrations of secondary
compounds such as tannin and saponin are good candidates for reducing ruminal
methanogenesis (Teferedegne, 2000).
Therefore
altering dietary formulation can have great impacts on environmental
performance from dairy operations.
Many studies
have reported that feeding forages containing tannin decreases ruminal protein
degradation (Min et al., 2003) and also have the potential to reduce enteric
CH4 emissions (Carulla et al., 2005; Animut et al., 2008).
It was also
reported that saponins or saponin-like substances had the potential to suppress
the methane emission, reduces protozoa counts and change fermentation patterns
(Hristov et al., 1999). At appropriate dose tannin and saponin decreases
methane production and increases the efficiency of microbial protein synthesis
(Min et al., 2003).
Therefore
this experiment is planned to ascertain the effect of plants extracts such as
tannin and saponin from different species and their effect on methane are other
in vitro fermentation parameter.
1.2 Thesis
Objectives
The study of
tannin and saponin content of some species and its effect on in vitro rumen
fermentation have been designed with the following objectives.
To determine
the tannin, crude protein, cell wall and saponin, contents of some spices in
Edo State, Nigeria.
To determine
the effects of adding spices as additives on in vitro ruminal methane
production and other fermentation parameters.
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