54
EVALUATION OF TOTAL MIXED RATIONS
SUPPLEMENTED WITH EXOGENOUS
FIBROLYTIC ENZYMES AND / OR LIVE YEAST
CULTURE IN BUFFALO BULLS
By
P. RAVIKANTH REDDY B.V.Sc. & A.H.
THESIS SUBMITTED TO THE
SRI VENKATESWARA VETERINARY UNIVERSITY
IN PARTIAL FULFILMENT OF THE REQUIREMENTS
FOR THE AWARD OF THE DEGREE OF
MASTER OF VETERINARY SCIENCE
(ANIMAL NUTRITION)
IN THE FACULTY OF VETERINARY SCIENCE
DEPARTMENT OF ANIMAL NUTRITION
NTR COLLEGE OF VETERINARY SCIENCE,
GANNAVARAM
SRI VENKATESWARA VETERINARY UNIVERSITY
TIRUPATI – 517 502.
NOVEMBER, 2014
55
Certificate
P. RAVIKANTH REDDY has satisfactorily prosecuted the course
of research and that the thesis entitled “EVALUATION OF TOTAL
MIXED RATIONS SUPPLEMENTED WITH EXOGENOUS
FIBROLYTIC ENZYMES AND / OR LIVE YEAST CULTURE IN
BUFFALO BULLS” submitted is the result of original research work
and is of sufficiently high standard to warrant its presentation to the
examination. I also certify that the thesis or part thereof has not been
previously submitted by him for a degree of any University.
Date: (Dr. D. SRINIVAS KUMAR)
Major Advisor
Associate Professor
Department of Animal
Nutrition
NTR College of Veterinary
Science
Gannavaram – 521 102.
56
CERTIFICATE
This is to certify that the thesis entitled “EVALUATION OF TOTAL
MIXED RATIONS SUPPLEMENTED WITH EXOGENOUS FIBROLYTIC
ENZYMES AND / OR LIVE YEAST CULTURE IN BUFFALO BULLS”
submitted in partial fulfillment of the requirements for the degree of MASTER
OF VETERINARY SCIENCE for Sri Venkateswara Veterinary University,
Tirupati, is a record of the bonafide research work carried out by P.
RAVIKANTH REDDY, under our guidance and supervision. The subject of
the thesis has been approved by the Student’s Advisory Committee.
No part of the thesis has been submitted by the student for any other
degree or diploma. The published part has been fully acknowledged. All the
assistance and help received during the course of investigations have been
duly acknowledged by the author of the thesis.
(Dr. D. SRINIVAS KUMAR)
Chairman of the advisory Committee
Thesis approved by the Student Advisory Committee Chairman: Dr. D. Srinivas Kumar _____________________
Associate Professor Department of Animal Nutrition N.T.R College of Veterinary Science Gannavaram – 521 102
Member: Dr. E. Raghava Rao _____________________
Professor & Head Department of Animal Nutrition N.T.R College of Veterinary Science Gannavaram – 521 102
Member: Dr. K. Ananda Rao ______________________
Senior Scientist & Head Buffalo Research Station VR Gudem – 534 101
57
CONTENTS
Chapter No. Title Page No.
I INTRODUCTION 1- 3
II REVIEW OF LITERATURE 4 - 44
III MATERIALS AND METHODS 45 - 53
IV RESULTS 54 - 80
V DISCUSSION 81- 106
VI SUMMARY 107-111
LITERATURE CITED 112-127
58
TABLE OF CONTENTS
Chapter
No.
Title Page
I INTRODUCTION 1-3
II REVIEW OF LITERATURE
2.1 Effect of supplementation of exogenous fibrolytic
enzymes in ruminant rations
4
2.1.1 Mode of action of exogenous fibrolytic enzymes 4
2.1.2 Effect of exogenous fibrolytic enzymes
supplementation on in vitro digestibility
8
2.1.3 Effect of exogenous fibrolytic enzymes
supplementation on rumen fermentation pattern
12
2.1.3.1 Effect of EFE supplementation in Buffaloes 12
2.1.3.2 Effect of EFE supplementation in Cattle 12
2.1.3.3 Effect of EFE supplementation in Sheep and Goat 15
2. 1.4 Effect of exogenous fibrolytic enzymes
supplementation on nutrient digestibility and
nutritive value
17
2.1.4.1 Effect of EFE supplementation in Buffaloes 17
2.1.4.2 Effect of EFE supplementation in Cattle 19
2.1.4.3 Effect of EFE supplementation in Sheep and Goat 21
2.2 Effect of Yeast culture supplementation in ruminant
rations
24
2.2.1 Mode of action of live Yeast culture 24
2.2.2 Effect of Yeast culture supplementation on in vitro
digestibility
26
2.2.3 Effect of Yeast culture supplementation on rumen
fermentation pattern
28
2.2.3.1 Effect of Yeast culture supplementation in Buffaloes 28
2.2.3.2 Effect of Yeast culture supplementation in Cattle 29
59
2.2.3.3 Effect of Yeast culture supplementation in Shee.p
and Goat
31
2.2.4 Effect of exogenous fibrolytic enzymes
supplementation on nutrient digestibility and
nutritive value
34
2.2.4.1 Effect of Yeast culture supplementation in Buffaloes 34
2.2.4.2 Effect of Yeast culture supplementation in Cattle 36
2.2.4.3 Effect of Yeast culture supplementation in Sheep
and Goat
37
2.3 Effect of EFE and Yeast culture supplementation in
ruminants
41
2.3.1 Effect of EFE and Yeast culture supplementation on
invitro digestibility
41
2.3.2 Effect of EFE and Yeast culture supplementation on
rumen fermentation pattern
42
2.3.3 Effect of EFE and Yeast culture supplementation in
nutrient digestibility and nutritive value
44
III MATERIALS AND METHODS
3.1 Source of exogenous fibrolytic enzymes (EFE) and
live Yeast culture
45
3.2 Preparation of total mixed rations 46
3.3 In vivo studies 46
3.3.1 Selection of animals 46
3.3.2 Experimental design 46
3.3.3 Hosing and management 46
3.3.4 Feeding regimen 47
3.3.5 Weighing of animals 47
3.3.6 Metabolism trial 48
3.4 Rumen fermentation studies 48
3.5 In vitro studies 49
60
3.6 Collection of samples 49
3.6.1 Feed and feed residue 49
3.6.2 Faeces 49
3.6.3 Urine 50
3.7 Analytical methods 50
3.7.1 Proximate analysis 50
3.7.2 Analysis of cell wall constituents 51
3.7.3 Analysis of minerals 51
3.7.4 Rumen metabolic profile studies 51
3.8 Statistical analysis 52
IV RESULTS
4.1 Evaluation of total mixed rations 54
4.1.1 Chemical composition and cell-wall constituents of
Groundnut haulms
54
4.1.2 Chemical composition and cell-wall constituents of
total mixed rations
56
4.2 In vitro studies 56
4.2.1 In vitro digestibility of DM in total mixed rations 58
4.2.2 In vitro digestibility of CP in total mixed rations 58
4.2.3 In vitro digestibility of NDF in total mixed rations 58
4.2.4 In vitro digestibility of ADF in total mixed rations 58
4.3 Rumen fermentation studies 60
4.3.1 Rumen pH 60
4.3.2 Total volatile fatty acids (TVFA) 62
4.3.3 Ammonia nitrogen 62
4.3.4 Total nitrogen 64
4.3.5 TCA insoluble protein nitrogen 64
4.3.6 Residual nitrogen 67
61
4.3.7 Food and protozoal nitrogen 69
4.4 Metabolism studies in Murrah buffalo bulls 72
4.4.1 Apparent nutrient digestibilities 72
4.4.2 Nitrogen balance 74
4.4.3 Calcium balance 74
4.4.4 Phosphorus balance 77
4.4.5 Plane of nutrition of buffalo bulls 79
V DISCUSSION
5.1 Evaluation of total mixed rations 81
5.1.1 Chemical composition and cell-wall constituents of
Groundnut haulms 81
5.1.2 Chemical composition and cell-wall constituents of
total mixed rations 82
5.2 In vitro studies 83
5.2.1 In vitro DM digestibility (%) of total mixed rations 83
5.2.2 In vitro CP digestibility (%) of total mixed rations 84
5.2.3 In vitro NDF digestibility (%) of total mixed rations 85
5.2.4 In vitro ADF digestibility (%) of total mixed rations 86
5.3 Rumen fermentation pattern 87
5.3.1 Rumen pH 87
5.3.2 Total volatile fatty acids (TVFA) 89
5.3.3 Ammonia nitrogen (NH3-N) 90
5.3.4 Total nitrogen 92
5.3.5 TCA insoluble nitrogen 93
5.3.6 Residual nitrogen 95
5.3.7 Food and protozoal nitrogen 96
5.4 Metabolism studies in graded Murrah buffalo bulls 97
5.4.1 Apparent nutrient digestibilities 97
5.4.2 Nitrogen balance 99
62
5.4.3 Calcium balance 101
5.4.4 Phosphorus balance 102
5.4.5 Plane of nutrition 103
5.5 Conclusion 105
V SUMMARY 107
LITERATURE CITED 112
63
LIST OF TABLES
Table
No. Title Page No.
1 Chemical composition (% DMB) of Groundnut haulms 55
2 Chemical composition (% DMB) of total mixed rations
fed to buffalo bulls during the study
57
3 In vitro digestibility (%) of total mixed rations
supplemented with EFE and/or live yeast culture
59
4 Rumen pH of buffalo bulls fed total mixed rations with
exogenous fibrolytic enzymes and/or live yeast culture
61
5 TVFA concentration (MEq/L of SRL) in buffalo bulls
fed total mixed rations with exogenous fibrolytic
enzymes and/or live yeast culture
63
6
Ammonia nitrogen concentration (mg/100ml SRL) in
buffalo bulls fed total mixed rations with Exogenous
Fibrolytic Enzymes and/or live yeast culture
65
7 Total nitrogen concentration (mg/100ml SRL) in
buffalo bulls fed total mixed rations with Exogenous
Fibrolytic Enzymes and/or live yeast culture
66
8 TCA insoluble protein nitrogen (mg/100ml SRL) in
buffalo bulls fed total mixed rations with Exogenous
Fibrolytic Enzymes and/or live yeast culture
68
9
Residual nitrogen (mg/100ml SRL) in buffalo bulls fed
total mixed rations with Exogenous Fibrolytic Enzymes
and/or live yeast culture
70
10
Food and protozoal nitrogen (mg/100ml SRL) in
buffalo bulls fed total mixed rations with Exogenous
Fibrolytic Enzymes and/or live yeast culture
71
11 Apparent digestibility (%) of nutrients in buffalo bulls
fed total mixed rations with Exogenous Fibrolytic
Enzymes and/or live yeast culture
73
12 Nitrogen utilization in buffalo bulls fed total mixed
rations with Exogenous Fibrolytic Enzymes and/or live
yeast culture
75
64
13 Calcium utilization in buffalo bulls fed total mixed
rations with Exogenous Fibrolytic Enzymes and/or live
yeast culture
76
Table
No. Title Page No.
14 Phosphorus utilization in buffalo bulls fed total mixed
rations with Exogenous Fibrolytic Enzymes and/or live
yeast culture
78
15 Plane of nutrition of buffalo bulls fed total mixed
rations with Exogenous Fibrolytic Enzymes and/or live
yeast culture
80
65
Acknowledgements
I would like to start with the person, who made the biggest difference in my
life, my mentor, Dr. D. Srinivas Kumar, Associate Professor, Department of Animal
Nutrition, NTR College of Veterinary Science, Gannavaram. This work would not
have been possible without his guidance, support and encouragement. Under his
guidance I successfully overcame many difficulties and learned a lot. Despite of his
busy schedule, he used to review my thesis progress, give his valuable suggestions
and made corrections. His unflinching courage, perseverance and dedication towards
the work will always inspire me.
It is with sincerest gratitude that I acknowledge the support and help of my
Professor Dr. E. Raghava Rao, Professor & Head, Department of Animal Nutrition,
NTR College of Veterinary Science, Gannavaram. I consider it an honor and
fortunate to work under his scholarly able and timely guidance.
I owe my deepest gratitude to Dr. K. Ananda Rao, Senior Scientist & Head,
Buffalo Research Station, VR Gudem for his kind concern and consideration and for
his patience and steadfast encouragement to complete this study.
I express my profound sense of reverence to Dr. T. Janardhan Reddy, Rtd.
Professor, Dept. of Animal Nutrition, who was abundantly helpful and patience
enough to offer valuable suggestions during the course of my study and research.
I would like thank Dr. K. Raja Kishore, Asst. Professor, Dept. of Animal
Nutrition, for his contribution by stimulating suggestions and encouragement during
the course of study and lab work and D. P. Ramesh Babu, ,Asst. Professor, Dept. of
Animal Nutrtion, NTR College of Veterinary Science, Gannavaram for his nice
interaction and valuable suggestions in completing my thesis.
It gives me great pleasure in expressing my deepest gratitude to Dr.
Nagaraja Kumari, Dr. Sowjanya Lakshmi, Dr. Rajamma, Dr.Swarna
Venkateswarlu Dr. Basava Reddy, Dr. Iqbal hyder, Dr. Chaitanya and Dr.
Vasantha for their valuable suggestions regarding the thesis and my future plannings.
I am very much thankful to my dearest colleagues Dr. M. Kiran Kumar, Dr.
B. Vamsidhar and Dr. Harish Khanna for their nice company and cooperation.
66
I would like to acknowledge my seniors Dr. V.Subba Reddy, Dr.
Rosi Reddy, Dr. Bhaskara Rama Raju, Dr. B. Suresh Babu, Dr. Srinivas Reddy and
Dr. Sandeep Reddy for their moral support and motivation which drives me to get my
best.
I am fortunate enough to have such friends like Dr. Nagendra
Reddy, Dr. Hemanth Kumar, Dr. Chaitanya Shankar and Dr. B.Obula Reddy, Dr.
Karthik Reddy, Dr. Veena Dhanekula.
I would like to acknowledge my appreciation to my juniors Rama Chandra
Reddy, Chintayya, Praharsha, Sridhar Reddy, Venkata Krishna, Praveen Reddy,
Prabhakar Reddy, Seshi Reddy and Avula Reddy for helping me during the work and
also for providing good companionship.
I extend my special thanks to Rajendra Prasad, B.V. Ramesh, K..Jogi Raju,
Smt. P.Sujatha, L. Venkatacharyulu, K. Koteswara Rao and Sathish of Dept. of
Animal Nutrition for their help and cooperation during the course of my study and
research.
I am very much thankful to Sri. K.S.R. Vithal Assoc. Professor & Head,
Library and the library staff for their extensive and timely help in collecting the
literature.
I extend my sincere thanks to Sri Venkateswara Veterinary University,
Tirupati for providing facilities to carry out my M.V.Sc research work in time.
Last but not the least, I have to thank my father P. Venkata Reddy and my
mother P. Sharadha Devi for their love and support through out my life and without
whom I can’t imagine even my existence. It is a great pleasure to thank my sweetest
sister P. Meenakshi Devi and brother P. Sagar Reddy with whom my early journey of
life was so happy, cheerful and progressive.
Ravi Kanth Reddy. P…
67
DECLARATION
I, Mr. P. RAVI KANTH REDDY hereby declare that the thesis
entitled “EVALUATION OF TOTAL MIXED RATIONS
SUPPLEMENTED WITH EXOGENOUS FIBROLYTIC ENZYMES
AND / OR LIVE YEAST CULTURE IN BUFFALO BULLS” submitted
to SRI VENKATESWARA VETERINARY UNIVERSITY for the
Degree of MASTER OF VETERINARY SCIENCE is a result of original
research work done by me. It is further declared that the thesis or any
part thereof has not been published earlier in any manner.
Date: (P. RAVI KANTH REDDY)
68
Name of the author : P. RAVIKANTH REDDY
Title of thesis : EVALUATION OF TOTAL MIXED RATIONS
SUPPLEMENTED WITH EXOGENOUS
FIBROLYTIC ENZYMES AND / OR LIVE
YEAST CULTURE IN BUFFALO BULLS
Degree to which it is
submitted : MASTER OF VETERINARY SCIENCE
Faculty : VETERINARY SCIENCE
Department : ANIMAL NUTRITION
Major advisor : Dr. D. SRINIVAS KUMAR Ph.D.,
Associate Professor
Department of Animal Nutrition
N.T.R. College of Veterinary Science
Gannavaram – 521 102
University : SRI VENKATESWARA VETERINARY UNIVERSITY
Tirupati – 517 502
Year of submission : September, 2014
ABSTRACT
In 4 x 4 LSD, four fistulated graded Murrah buffalo bulls (Avg. B.W.
377.05 ± 43.36 kg) were randomly allotted to four dietary treatments viz.
groundnut haulms based TMR with R: C ratio of 70: 30 (T1), T1 supplemented
with exogenous fibrolytic enzymes (EFE) @ 15 g/animal/day (T2), T1
supplemented with yeast culture @ 10 g/animal/d (T3) and T1 supplemented
with EFE @ 15 g/animal/day and yeast culture @ 10 g/animal/d (T4) and
evaluated for their effect on in vitro digestibility, rumen fermentation pattern,
mineral balances and nutrient utilization in buffalo bulls. The in vitro
digestibility (%) of DM, CP, NDF and ADF were lower (P<0.01) in T1 when
compared to T2, T3 or T4. Further, the in vitro digestibility (%) of DM, CP,
NDF and ADF increased linearly from T2 to T4. However, no significant
69
(P>0.05) differences were observed between T2 and T3, T2 and T4 and T3 and
T4.
Rumen fermentation studies conducted in fistulated buffalo bulls
revealed that rumen pH values were highest at 0 h and declined to minimum by
4 h post feeding, while TVFA, NH3-N, and N fractions reached peak at 4 h post
feeding and later followed a decreasing trend in all the treatments. The present
study indicated that supplementation of EFE in TMR (T2) had no effect
(P>0.05) on rumen pH and food and protozoal N concentration while it
increased (P<0.01) the TVFA, NH3-N and other N fractions when compared to
T1. Further, yeast culture supplementation in TMR (T3) increased (P<0.01)
rumen pH, TVFA, NH3-N, total N, TCA-insoluble N and residual N while it
had no effect (P>0.05) on food and protozoal N in buffalo bulls. Furthermore,
supplementation of EFE and/or live yeast culture in TMR (T4) increased
(P<0.01) rumen pH, TVFA, NH3-N and N fractions in buffalo bulls as
compared to the control group.
The digestibility coefficients (%) of gross nutrients and fibre fractions
increased linearly from T1 to T4 but the differences were not significant
(P>0.05). The study indicated that supplementation of EFE and/or live yeast
culture in TMR had no effect (P>0.05) on the digestibility of gross nutrients
and fibre fractions. All the buffalo bulls were in positive N, Ca and P balance.
Further, supplementation of EFE and/or live yeast culture in TMR had no
effect (P>0.05) on N, Ca and P retentions expressed as either g/d or % intake or
% absorbed. The average DMI of buffalo bulls expressed as kg/d or as % BW
was comparable among the treatments. Supplementation of EFE and/or live
yeast culture in TMR had no effect (P>0.05) on DCP and TDN contents
expressed as % in the diet consumed or as kg/d. Furthermore, the DM, DCP,
TDN and ME intakes per kg W0.75
were similar among the treatments and were
higher than the values recommended by ICAR (1998) and Kearl (1982)
standards.
70
Based on the results of the present study, it is concluded that
supplementation of EFE and/or live yeast culture to groundnut haulms based
TMR resulted in improved digestibility of nutrients in vitro while in vivo
studies conducted in buffalo bulls revealed no affect on DM intake and on
digestibility of gross nutrients and fibre fraction
71
LIST OF ABBREVATIONS
% : Percent
± : Plus or Minus
g : Grams
g/d : Grams/day
Kg : Kilogram
ADF : Acid detergent fibre
ADL : Acid detergent lignin
AFRC : Agricultural Food and Research Council
AOAC : Association of Official Analytical Chemists
Ca : Calcium
CF : Crude fibre
CP : Crude protein
DCP : Digestible crude protein
DE : Digestible energy
DM : Dry matter
DMB : Dry matter basis
DMI : Dry matter intake
EE : Ether extract
EFE : Exogenous fibrolytic enzymes
ICAR : Indian Council of Agricultural Research
IVDMD : In vitro dry matter digestibility
IVCPD : In vitro crude protein digestibility
IVNDFD : In vitro neutral detergent fibre digestibility
IVADFD : In vitro acid detergent fibre digestibility
LSD : Latin square design
ME : Metabolizable energy
MEq : Milli-equivalent
N : Nitrogen
NDF : Neutral detergent fibre
NFE : Nitrogen free extract
NH3-N : Ammonia nitrogen
72
OM : Organic matter
P : Phosphorus
R: C ratio : Roughage : Concentrate ratio
SRL : Strained rumen liquor
TA : Total ash
TCA : Trichloro acetic acid
TDN : Total digestible nutrients
TMR : Total mixed ration
Total-N : Total nitrogen
TVFA : Total volatile fatty acids
W kg0.75
: Metabolic body weight
73
CHAPTER – I
INTRODUCTION
The scarcity of green fodder and high cost of conventional feed
ingredients have prompted the researchers to improve the feeding value of poor
quality crop residues. Among the various agricultural poor quality crop
residues, groundnut (Arachis hypogea) haulms (GNH) are abundantly available
in the tropical region of the country. Groundnut, a principal oil seed crop of
India, is grown on about 7.6 million hectares with a production of 7.8 million
tons of nuts in shell. The crop residue left after harvesting the pods is known as
groundnut haulms, and is an excellent source of crude protein, ether extract,
calcium and phosphorous with 5.7% DCP and 55.8% TDN (Shukla et al.,
1985). However, it is high in fibre which limits its use as sole feed for
ruminants. Further, the high fiber content prevents the access of ruminal
hydrolytic enzymes to cellulose and hemicelluloses (Tan et al., 1995).
During recent years, yeast culture and fibrolytic enzymes have been
used to improve the nutritive value and utilization efficiency of poor quality
roughages. Fibrolytic enzymes and yeast culture supplementation in ruminant
diets can increase DMI, production performance, cellulose degradation, and
nutrient digestibility (Kung et al., 1997). Addition of cellulolytic enzymes to
forage at the time of ensiling can aid in the digestion of cell wall
polysaccharides, resulting in greater digestibility and increased intake
74
(Colombatto et al., 2003). When an exogenous fibrolytic enzyme was applied
to the concentrate portion of diet or sprayed onto the total mixed ration before
feeding, the nutrient digestibility in the total digestive tract was increased for
early lactating cows (Yang et al., 2000). Similarly, using the exogenous
fibrolytic enzyme in high grain diets or in grass forages improved fiber
digestion and grain utilization both in situ and in vivo (Feng et al., 1996;
Krause et al., 1998).
Yeast culture (Saccharomyces cerevisiae) has been extensively used as a
dietary supplement in ruminants. The benefits associated with S. cerevisiae
include increased DM and NDF digestion (Carro et al., 1992; Plata et al.,
1994), increased initial rate of fiber digestion (Williams et al., 1991), improved
in situ CP and NDF degradation and microbial efficiency (Olson et al., 1994),
and increased DMI and milk production (Williams et al., 1991; Piva et al.,
1993; Kung et al., 1997). In vitro studies have also shown that yeast culture
favorably altered the mixed ruminal microorganism fermentation and
stimulated cellulose digestion by pure cultures of predominant ruminal bacteria
(Callaway and Martin, 1997). In recent years, attempts have also been made to
feed dairy animals with a composite of several types of probiotic preparations
(Erasmus et al., 2005), assuming their synergistic effect on the productivity and
health of animals. However, only few comparative studies were conducted on
the effect of addition of live yeast culture and / or exogenous fibrolytic
enzymes on productivity and health of dairy animals (Can et al., 2007; Tang et
al., 2008; Lopuszanska – Rusek and Bilik, 2011).
75
Hence, the present experiment was designed to investigate the effect of
direct addition of fibrolytic enzymes preparation and/or yeast culture to
groundnut haulms based TMR in graded Murrah buffalo bulls with the
following objectives:
1. To study the effect of supplementing exogenous fibrolytic enzymes
and/or live yeast culture to total mixed rations on in vitro digestibility of
nutrients.
2. To study the effect of supplementation of exogenous fibrolytic enzymes
and/or live yeast culture to total mixed rations on rumen fermentation
pattern in bulls.
3. To study the effect of feeding total mixed rations supplemented with
exogenous fibrolytic enzymes and/or live yeast culture on nutrient
utilization in bulls.
76
CHAPTER - II
REVIEW OF LIETRATURE
2.1 EFFECT OF SUPPLEMENTATION OF EXOGENOUS
FIBROLYTIC ENZYMES IN RUMINANT RATIONS
In recent years, supplementation of exogenous fibrolytic enzymes (EFE)
as feed additives has received considerable attention. It has been demonstrated
that EFE work in synergy with the endogenous rumen microbial enzymes to
enhance the digestibility and nutritive value of high fibrous diet (Morgavi et
al., 2000), thereby increasing the economic benefits for the farmer. The
research conducted using EFE in ruminants, the possible mode of action and
the effect of supplementation of EFE in the rations of ruminants on various
parameters are reviewed here.
2.1.1 MODE OF ACTION OF EXOGENOUS FIBROLYTIC ENZYMES
The probable modes of action of exogenous feed enzymes in ruminants
can be categorized as shown below.
2.1.1.1 Pre Consumption Effects
The EFE are most effective when applied in liquid form onto dry feed
prior to ingestion (Kung et al., 2000; Beauchemin et al., 2003). This may
77
partially digest feed or weaken cell wall barriers that limit microbial digestion
in the rumen. The EFE releases reducing sugars from the feedstuffs before feed
consumption (Hristov et al., 1996) which arises at least partially from the
solubilisation of NDF and ADF (Hristov et al., 1996; Krause et al., 1998;
Gwayumba and Christensen, 1997). This may therefore increase available
carbohydrates in the rumen (Yang et al., 2000) and also enhance the rapid
microbial attachment and growth (Forsberg et al., 2000). The alteration of feed
structure, due to the partial solubilisation of cell wall before feeding, is more
likely to increase feed degradation in the rumen (Beauchemin et al., 2004).
Further, treating feed with EFE prior to feed consumption will improve the
binding of enzyme to feed particles compared to its direct addition in the
rumen. This is reported to increase the resistance of EFE to proteolysis in the
rumen (Morgavi et al., 2001; Beauchemin et al., 2003).
2.1.1.2 Ruminal Effects
McAllister et al. (2001) reported that in the rumen, the EFE may
hydrolyse feed directly or work synergistically with ruminal microbes to
enhance feed digestion. Hristov et al. (1998) reported that application of EFE
@ 12 mg/g resulted in increased xylanase and cellulase activities in the rumen
by 32% and 11%, respectively. Similarly, Wallace et al. (2001) studied the
stability of EFE in the rumen fluid and reported that addition of EFE to the diet
@ 1.5 mg/g resulted in increased xylanase activity by 5% and cellulase activity
by 15%. These two studies indicated that EFE were actively stable and
78
hydrolyze the feed in the rumen fluid. Evidences of the stability of EFE in the
rumen indicated the substantial synergism between EFE and enzymes secreted
in the rumen and that the net combined hydrolytic activity in the rumen is much
higher than estimated from single sources (Beauchemin et al., 2004). This
positive synergy was reported as a result of an increased in vitro gas
production, total VFA, true degradability of substrate DM and decreased
methane production (Giraldo et al., 2008a). Morgavi et al. (2000) speculated
that the synergy is likely a significant mechanism by which enzyme additives
improve feed digestion. In sub rumen conditions (pH > 5.9) resulted from using
high fermentable diet, EFE effectiveness was considered to be reduced
compared to its effectiveness at higher rumen pH conditions (Beauchemin et
al., 2004). Yang et al. (2002) revealed that the effect of EFE rather than
enhanced microbial activity was the reason for improved fibre digestion in the
rumen during sub-optimal rumen conditions.
Another benefit of EFE application in ruminants is the indirect increase
of attachment and numbers of cellobiose- and glucose- utilizing bacteria in the
rumen (Nsereko et al., 2002). Further, enzyme extracts from Aspergillus oryzae
increased the number of rumen bacteria (Newbold et al., 1992) and work
synergistically with extracts from rumen microorganisms to enhance release of
soluble sugars from hay (Newbold, 1995). Similarly, Giraldo et al. (2008b)
reported that treating high-forage diet with EFE stimulated the in vitro numbers
of microbes and enhanced the fibrolytic activity. The microbial stimulation can
increase the availability of substrate as a result of an improved cell wall
79
digestion and may accelerate the digestion of newly ingested feedstuffs
(Beauchemin et al., 2004) thus leading to increased synergy between EFE and
enzymes of the rumen. Further, the stimulation of total microbial numbers by
EFE can result in increased microbial biomass thus increasing the supply of
metabolizable protein to the small intestine (Yang et al., 1999). Thus,
improvements in digestibility due to an increased hydrolytic activity can also
be attributed to an increased digestion of non-structural components in addition
to increased fibre digestion (McAllister et al., 2001).
2.1.1.3 Post Ruminal Effects
EFE not only increase the fibrolytic activity in the rumen but also
improve fibrolytic activity in the small intestine (Hirstov et al., 2000). In the
small intestine, EFE appear to survive for a sufficient period of time with
sufficient effects on substrate particles when applied to wet feeds and
concentrate premix (Morgavi et al., 2001; Beauchemin et al., 2004). This may
improve nutrient absorption by hydrolyzing substrates that rapidly escape
digestion in the rumen.
It is also reported that EFE work synergistically with microbes even in
the large intestine (Beauchemin et al., 2004). Hirstov et al. (2000) reported that
the xylanase activity in the faeces increased linearly with increasing levels of
enzyme supplementation. Further, feeding exogenous phytases and cellulases
to lactating cows resulted in improved digestibility of feed and reduced the
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faecal excretion of DM, NDF, N and P fractions (Knowlton et al., 2007).
Recently, Sajjad et al. (2008) suggested that EFE may improve feed digestion
within the rumen either by pre-treating the feed with EFE or by directly
increasing the fibrolytic activity into the rumen.
2.1.2 EFFECT OF EFE SUPPLEMENTATION ON IN VITRO
DIGESTIBILITY
Shojaeian and Thakur (2006) conducted a study to find out the effect of
incorporating isobutyrate and EFE in urea and NaOH treated wheat straw on in
vitro degradability. They reported that different levels of EFE or BCFA
supplementation improved IVDMD and IVNDFD of wheat straw, 4% urea
treated wheat straw and 3% NaOH treated wheat straw. Similarly, Balci et al.
(2007) conducted an in vitro study to evaluate the effect of supplementing
fibrolytic enzyme (Promote N.E.T) to either wheat straw or concentrate and
reported that wheat straw showed higher (P<0.05) in vitro DM, OM and NDF
digestibility in the treated group compared to the control, whereas concentrate
feed did not show the same differences (P>0.05).
Miachieo and Thakur (2007) conducted a study to assess the effect of
supplementing a mixture of cellulase and xylanase (1:1) @ 1.5 or 3.0 g/kg DM
on in vitro digestibility of diet containing wheat straw, concentrate mixture and
green fodder in 40:40:20 ratios. They reported that the IVDMD, IVOMD and
IVNDFD were higher (P<0.05) at E1.5 and E3.0 levels, both at 24 h and 48 h
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post-incubation compared to E0 but no significant difference was observed
between E1.5 and E3.0. Later, Shojaeian and Thakur (2007) conducted in vitro
studies on untreated or 4% urea treated wheat straw and concentrate mixture
ground to 1 mm screen size and mixed in R: C ratio of 70: 30, 60: 40 and 50:
50 (on DMB), supplemented at 1.5 and 3 g/kg DM of enzyme mixture
containing equal parts of cellulase and xylanase (EFE). Results revealed no
difference (P>0.05) in IVDMD and IVNDFD between 50: 50 and 60: 40 R: C
ratios and between both levels of EFE supplementation. However, both
IVDMD and IVNDFD were higher (P<0.01) on 60: 40 and 50: 50 than 70: 30
R: C ratio in the substrate.
Thakur et al. (2008) conducted an in vitro experiment to identify
promising EFE, cellulase and xylanase combinations and their optimum level
of supplementation for improving digestibility of TMR. Three enzyme
mixtures E1, E2 and E3 [containing cellulase (4000 IU/g) and xylanase (7990
IU/g)] were added at 1.5, 3.0 and 6.0 g/kg DM to three substrates S1, S2 and S3
[containing concentrate: wheat straw: green fodder (sorghum) in 25:60:15,
40:45:15 and 60:25:15 proportions on DMB, respectively]. IVDMD and
IVNDFD of all substrates at 48 h were improved on addition of EFE in E1, E2
and E3 proportions at 1.5 and 3.0 g/kg DM of TMR while both IVDMD and
IVNDFD declined (P<0.05) on addition of 6.0 g EFE/kg DM of TMR.
Ganai et al. (2011) carried out in vitro studies to evaluate the effects of
EFE supplemented to bajra straw @ 0, 1, 2, 3 and 4 g/kg substrate DM on
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degradability of DM and NDF using goat rumen liquor. Results indicated
significant (P<0.01) improvement in nutrient degradability of DM and NDF in
bajra straw showing maximum values at enzyme level of 2 g/kg DM.
Thakur and Shelke (2011) studied the effect of different periods of
storage and temperatures of TMR containing EFE on enzyme activity and in
vitro digestibility. They reported that there was no adverse effect of storage
(upto 60 days) and heating (up to 800C) of TMRs containing fibrolytic enzymes
on cellulase and xylanase activities and in vitro fibre digestibility. In another
experiment, Yancy Mary Issac et al. (2011) included Sorghum stover/GN
haulms with and without enzyme treatment in complete ration for sheep and
compared for their efficiency on DMD through in vitro studies using rumen
liquor from sheep. The in vitro DMD of enzyme treated complete ration (53.60
± 3.57) was significantly higher (P<0.05) compared to in vitro DMD (37.2 ±
2.02) of untreated complete ration .
Bhasker et al. (2012) had undertaken in vitro studies to evolve
appropriate fibrolytic cocktail enzyme comprising of cellulase, xylanase and β-
D gluconase for sorghum stover based on IVDMD using rumen liquor from
cattle maintained on sorghum stover. The study indicated that EFE have
potential to increase the IVDMD of sorghum stover and that the enzyme
cocktails (cellulase - xylanase - β-D glucanase, IU/g) at the concentration of
32000 – 25600 - 400 or 38400 – 12800 - 100 was optimum for enhancing
nutrient utilization from sorghum stover for large ruminants.
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Azzaz et al. (2013) conducted an in vitro experiment using rumen fluid
from rams fed berseem hay ration to study the effect of supplementing dried
sugar beet pulp with laboratory produced fibrolytic enzyme (Asperozym) and
commercial fibrolytic enzymes source (Tomoko®) at 3 levels (0, 1, 1.5 and 2
g/kg DM) on IVDMD and IVOMD. Results indicated that increasing the
Asperozym and Tomoko®
supplementation levels up to 2 g/kg DM exhibited
the highest (P<0.05) values of IVDMD and IVOMD.
Bhasker et al. (2013c) conducted in vitro studies to develop an
appropriate fibrolytic cocktail enzyme comprising of cellulase, xylanase and β-
D-glucanase for maize stover with an aim to increase its nutrient utilization in
sheep. They reported that cellulase and xylanase added individually to ground
maize stover at an increasing dose rates (0, 100, 200, 400, 800, 1600, 3200,
6400, 12800, 25600, 32000, 38400 and 44800IU/g DM) increased (P<0.01) the
in vitro dry matter digestibility. Recently, Rajamma (2013) conducted an in
vitro study to evaluate the effect of EFE supplementation on the digestibility of
TMRs containing different R : C ratio. Results indicated that in vitro
digestibility (%) of DM, CP, NDF and ADF were higher (P<0.01) in TMR with
R : C ratio of 60 : 40 (T1) when compared to that with 70 : 30 (T3). Further,
supplementation of EFE in TMRs irrespective of R : C ratio increased the in
vitro digestibility (%) of DM, CP, NDF and ADF.
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2.1.3 EFFECT OF EFE SUPPLEMENTATION ON RUMEN
FERMENTATION PATTERNS
2.1.3.1 Effect of EFE supplementation in Buffaloes
Singh and Das (2009) conducted an experiment to study the effect of
fibrolytic enzyme (Abizyme forte) treatment of wheat straw on rumen
fermentation in buffalo calves. Results indicated that the rumen pH, NH3-N and
total nitrogen concentrations were not affected by the enzyme treatment while
the TVFA concentration was significantly (P<0.05) higher (84.83 m mol/L) in
enzyme treated group as compared to control (74.50 m mol/L).
Thirty multiparous lactating buffaloes were fed roughage with three
levels of fiber (high, medium, low) with or without fibrolytic enzymes. Results
indicated that the pH value increased (P<0.05) while TVFA and NH3-N
concentrations decreased (P<0.05) with increasing dietary fibre content.
Further, the pH value and NH3-N concentration were lower (P<0.05) but
TVFAs concentration was higher (P<0.05) with fibrolytic enzyme
supplementation (Gaafar et al., 2010).
In 4 x 4 LSD, four fistulated buffalo bulls were randomly allotted to four
dietary treatments viz. maize stover based TMR with R: C ratio of 60: 40 (T1),
T1 supplemented with EFE @ 15 g/animal/day (T2), maize stover based TMR
containing R: C in 70: 30 ratio (T3) and T3 supplemented with EFE @ 15
g/animal/day (T4). Results revealed that rumen pH was lower (P<0.01) while
the concentration of TVFA, NH3-N and N fractions were higher (P<0.01) in T1
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when compared to T3. Further, supplementation of EFE in TMRs irrespective
of R: C ratio decreased (P<0.01) rumen pH and increased (P<0.01) the
concentrations of TVFA, NH3-N and N fractions in buffalo bulls (Rajamma et
al., 2014).
2.1.3.2 Effect of EFE supplementation in Cattle
In 4 x 4 LSD, 4 rumen cannulated steers were allotted to 4 dietary
treatments viz. Enzyme 1 (cellulase; E1) applied to fresh forage, then dried; E1
applied to wilted forage, then dried; a combination of enzyme 1 and enzyme 2
(xylanase; E2) in 50:50 combination (E1E2) applied to dry forage immediately
before feeding (E-dry) and untreated forage (Control). All forage treatments
were harvested as dry hay. Results indicated that rumen pH, NH3-N and TVFA
were not altered (P>0.10) by dietary treatment (Feng et al., 1996).
Baah et al. (2005) fed non lactating Holstein cows with TMR containing
50 % rolled barley grain and 50 % orchard grass hay treated with either water
(Control), 0.2 % (vol/wt) Tween 80, 0.2 % (vol/wt) hydrolytic enzyme or 0.2 %
hydrolytic enzyme plus 0.2 % Tween 80. They reported that rumen pH and
TVFA concentrations were not affected (P>0.05) by enzyme treatments.
Similarly, Balci et al. (2007) observed no change in rumen pH (6.19 ± 0.13 vs.
6.20 ± 0.07 in control and enzyme treated groups, respectively) in steers fed on
wheat straw based diets supplemented with Promote N.E.T @ 60 g/day/steer
mixed in concentrate.
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Alvarez et al. (2009) conducted an experiment by allotting 6 rumen
cannulated steers fed oat straw-wheat midlings based diets to three treatments
viz. Control, Fibrozyme (sprayed @ 2 g/kg DM) and Promote N.E.T (sprayed
@ 3 ml/kg DM) to evaluate their effect on rumen fermentation. Results
indicated that both the enzymes increased rumen pH values as compared to
control while there was no effect on TVFA concentration. Later, Gado et al.
(2009) studied the effects of a mixture of exogenous enzymes (ZADO®) from
anaerobic bacteria on ruminal fermentation in cows fed TMRs containing corn
silage and concentrate mixture in 70: 30 ratio. Twenty lactating multiparous
Brown Swiss cows were randomly assigned into 2 groups and fed TMR with or
without addition of 40 g/cow/d of enzymes for 12 weeks. Results indicated that
supplementation of enzymes increased (P<0.05) rumen ammonia N while
rumen pH decreased (P>0.05) compared to control.
Bilik and Lopuszanska-Rusek (2010) conducted an experiment using 12
cows divided into 2 groups of 6 each and fed TMRs with or without fibrozyme.
Results revealed significant increase (P<0.01) in TVFA content while there
was no effect on pH and NH3-N concentration between the groups. In another
study, Bassiouni et al. (2010) randomly assigned 30 multiparous lactating
Friesian cows into 6 groups (5 in each group) and fed experimental rations
consisting (on DMB) of 60% concentrate feed mixture (CFM) + 40% berseem
hay without (BH) or with fibrozyme (BH-E), corn silage without (CS) or with
fibrozyme (CS-E) and rice straw without (RS) or with fibrozyme (RS-E) added
@ 1 g/kg DM. Results revealed that cows fed CS-E ration had significantly
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(P<0.05) the lowest pH value and the highest NH3-N concentration, while those
fed BH ration revealed the highest NH3-N concentration. The pH
value
decreased, but TVFA and NH3-N concentrations increased gradually until 4 h
after feeding and then declined at 6 h.
Phakachoed et al. (2012) conducted a 3 x 3 LSD study by allotting three
fistulated non-lactating dairy cows to three treatments viz. control, fed 10 g
xylanase/cow/d and 20 g xylanase/cow/d to evaluate the effect of xylanase
product (Porzyme) on rumen fermentation. The basal diet comprised of 3 kg/d
of concentrate containing 17 % CP together with ad libitum rice straw. Results
indicated that rumen pH, NH3-N and TVFA concentration at each hour of
incubation were unaffected by enzyme supplementation.
2.1.3.3 Effect of EFE supplementation in Sheep and Goat
Pinos-Rodriguez et al. (2002) conducted a study by randomly assigning
four ruminally cannulated lambs on four diets viz. alfalfa hay, alfalfa hay +
enzyme, rye grass hay and rye grass hay + enzyme to study the effect of
directly fed exogenous fibrolytic enzyme on rumen fermentation. Results
indicated that the enzyme had no effect on rumen pH and NH3-N while the
concentration of TVFA increased (P<0.05) at 3 h and 6 h but was not affected
12 h post feeding. Later, in another study, Pinos-Rodriguez et al. (2008)
reported that addition of fibrozyme @ 2 g/Kg DM of TMR of lambs containing
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different forage : concentrate ratios had no effect on rumen pH, NH3-N and
TVFA concentration irrespective of F : C ratio..
In a crossover design experiment, 6 rumen fistulated Merino sheep were
fed mixed grass hay: concentrate (70 : 30 on DMB) diet to evaluate the effects
of an exogenous fibrolytic enzyme preparation (12 g/d; ENZ) delivered directly
into the rumen on rumen fermentation. Results revealed that supplementation
with ENZ did not affect rumen pH or concentrations of NH3-N and total VFA
(Giraldo et al., 2008b).
Avellaneda et al. (2009) carried out a repeated LSD within a 2×2
factorial arrangement (i.e., enzyme, 0 and 3 g/d/lamb; Guinea grass, 35 and 90
d) to determine the effects of EFE on ruminal degradation of Guinea grass
(Panicum maximum var mombasa) hay cut at 35 and 90 days of growth using
four Suffolk lambs (74.0 ± 2.01 kg BW) fitted with ruminal and duodenal
cannulae. Results indicated that enzyme supplementation had no effect on
ruminal pH, ammonia N and TVFA concentration.
In a crossover design experiment, 6 rumen fistulated Merino sheep were
fed mixed grass hay and concentrate in 70 : 30 ratios with or without direct fed
EFE (12 g/d) to study the effect on ruminal fermentation. Results indicated that
EFE supplementation had no affect (P>0.05) on ruminal pH and concentrations
of NH3-N at any sampling time (Giraldo et al., 2009). Later, Ganai et al. (2011)
conducted an in vitro study to evaluate the effects of EFE (2 g/kg DM) on
rumen fermentation characteristics in complete feed containing bajra straw and
89
concentrate mixture in 60 : 40 ratios. Results revealed that supplementation of
EFE improved (P<0.01) significantly the TVFA but did not alter the rumen pH,
total nitrogen and ammonia N compared to control.
Later, Bhasker et al. (2013c) conducted a trial on 12 rams by feeding 50
% maize stover based TMR supplemented with or without enzyme (cellulase-
xylanase, 12,800 - 12,800 IU/g) to study the effect on rumen fermentation
pattern. Results indicated that TVFA (P<0.01) and NH3-N (P<0.05)
concentration was higher in enzyme supplemented group while no effect was
observed on pH and total N concentration. In another study, twenty Pelibuey
lambs were randomly assigned to control or to one of three enzyme treatments
to evaluate their effects on rumen fermentation. The study indicated that the
fibrolytic enzyme extracts supplementation had no effect on pH, NH3-N and
TVFA concentration in lambs (Torres et al., 2013).
2.1.4 EFFECT OF EFE SUPPLEMENTATION ON NUTRIENT
DIGESTIBILITY AND NUTRITIVE VALUE
2.1.4.1 Effect of EFE supplementation in Buffaloes
EL-Kady et al. (2006) conducted an experiment to investigate the effect
of an enzyme mixture containing cellulase, xylanase, α-amylase and pectinase
activity in fermented Sugar Beat pulp (SBP) in buffalo calves fed on rations
containing concentrate feed mixture (CFM) at 2 % body weight and Pearl
Millet ad libitum. To the CFM, SBP was added at 0 (R1), 0.2 (R2), 0.4 (R3), 0.6
90
(R4) % w/w and fed to 4 groups of buffalo calves (4 in each group). Results
revealed that supplementation of enzyme has no affect on feed intake while the
digestibility of CP, EE, NFE and TDN values were significantly (P<0.05)
higher for all enzyme supplemented groups over control group.
Singh and Das (2009) reported that the digestibility coefficients for DM,
OM, NDF, ADF and CF were higher (P<0.05) in buffalo calves fed enzyme
treated diet compared to control. Further, there was no affect on DMI, DCP and
TDN intake (kg/d) or N balance (g/d). Later, Gaafar et al. (2010) reported that
the digestibility of DM, OM, CP, EE and NFE and nutritive values decreased
(P<0.05) while CF increased (P<0.05) in lactating buffaloes with increasing
fibre content in the diet. Similarly, the digestibility of all nutrients and nutritive
values increased (P<0.05) with supplementation of fibrolytic enzymes. Further,
the intake of DM, TDN, CP and DCP increased (P<0.05) with decreasing
dietary fibre content as well as with fibrolytic enzyme supplementation.
Fifteen mid lactating buffaloes (3months post parturition) were divided
into 3 groups of 5 animals each and were fed 3 different rations viz. control
ration (45 % CFM, 30 % corn silage, 15 % dried sugar beet pulp and 10 % rice
straw), R1 (control ration + Asperozym @ 2 g/Kg DM) and R2 (control ration +
Tomoko® @ 2 g/Kg DM). Results indicated that both Asperozym and
Tomoko® supplementation increased (P<0.05) the digestibilities of DM, OM,
CF, NFE, NDF in treatment groups compared to the control group (Azzaz et
al., 2013).
91
Recently, Rajamma (2013) studied the effect of supplementing EFE in
TMR containing different R : C on nutrient utilization in buffalo bulls. Results
revealed that supplementation of EFE in TMRs irrespective of R : C ratio
increased (P<0.01) the digestibility of CP, EE and CF while there was no effect
on the digestibility of other nutrients. Further, supplementation of EFE in
TMRs irrespective of R : C ratio had no effect (P>0.05) on intake of DCP
(kg/d), TDN (kg/d), DE (M cal) and ME (M cal).
2.1.4.2 Effect of EFE supplementation in Cattle
In switch back design, twelve lactating cows (6 early & 6 late) were
used to study the effect of fibrolytic enzyme formulation on N and P intake,
partitioning and excretion. Diets of early group contained 45 % forage while
late lactation group contained 61 % forage. Results indicated that the enzyme
did not significantly affect the apparent digestibility or excretion of N and P or
retention of these nutrients in the body tissue (Knowlton et al., 2002).
Wang et al. (2004) studied the effects of EFE supplementation (100
ml/kg DM) on ammoniated wheat straw using 32 Continental X British
crossbred beef cows (649.9±11.9 kg) in late gestation. They reported that
applying enzyme to ammoniated wheat straw increased (P<0.05) the
digestibilities of DM, OM and total tract N but did not affect DMI and ADF
digestibility. In another study, Baah et al. (2005) reported that total tract
digestibility coefficients of DM, nitrogen, NDF and ADF were not affected
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(P>0.05) by dietary treatments supplemented with fibrolytic enzymes in non
lactating Holstein cows.
In another study, Gado et al. (2009) reported the intake of DM and OM
was positively influenced (P<0.05) by EFE supplementation, and the
digestibility of all nutrients was higher (P<0.05) in the total tract of
supplemented cows, although the magnitude of the improvement varied among
nutrients, with the highest improvement in aNDFom and ADFom. Later, in a
cross over design experiment, Peters et al. (2010) studied the effect of feeding
a TMR applied with or without EFE product prior to feeding in multiparous
lactating Holstein cows on nutrient digestion. Results indicated that enzyme
supplementation had no affect on the apparent digestibility of DM, OM, NDF
and ADF.
Bassiouni et al. (2010) conducted an experiment by dividing lactating
Friesian cows into three groups fed different diets viz. CS-E ration (Corn silage
supplemented with Fibrozyme), BH-E ration (Berseem hay supplemented with
Fibrozyme) and RS ration (unsupplemented rice straw). Results revealed that
cows fed CS-E rations showed the highest DM and TDN intake, those fed with
BH-E rations had the highest DCP intake and those fed with RS rations
recorded the lowest intake (P<0.05). The BH-E rations showed the highest
digestibility coefficients of DM, OM, CP and CF and DCP value while the CS-
E rations showed the highest EE digestibility and TDN value.
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2.1.4.3 Effect of EFE supplementation in Sheep and Goat
Pinos-Rodriguez et al. (2002) reported that enzyme supplementation to
either alfalfa or rye grass in lambs resulted in increased intake of DM (P<0.01),
OM and CP (P<0.05) while there was no affect on NDF and ADF intakes.
Further, enzyme supplementation to alfalfa hay increased the digestibilities of
CP, hemicellulose (P<0.05) and NDF (P<0.01). Also, for both hays, the
enzyme improved N balance because lambs retained more N (P<0.05). Later,
Titi and Tabbaa (2004) carried out an experiment by dividing 10 lambs into
two groups viz. cellulase treated and control, to investigate the efficacy of
direct feeding of fibrolytic enzyme on nutrient digestibility. Results revealed
that treated lambs had higher (P<0.05) digestibilities for DM, OM, CF NDF
and ADF and also retained more (P<0.05) nitrogen in their bodies than lambs
control group.
Pinos-Rodriguez et al. (2008) reported that addition of Fibrozyme @ 2
g/Kg DM of TMRs of lambs containing different roughage concentrate ratios
had no affect either on feed intake, digestibility of DM and NDF or on N
balance. Similarly, Avellaneda et al. (2009) reported that there was no enzyme
effect on DM intake, N balance and total tract digestibilities in growing Suffolk
lambs fed guinea grass hay cut at 35 and 90 days.
Yancy Mary Issac et al. (2011) conducted a digestibility trial in sheep to
study the extent to which digestibility was influenced by treatment of sorghum
stover/GN haulms with NSP mixtures. They reported that the digestibility of
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enzyme treated sorghum stover/GN haulms were significantly (P<0.05) higher
than that of their untreated counterparts. On the other hand, Wahyuni et al.
(2012) studied the effect of supplementing the TMR containing oil palm frond
(OPF) silage with different levels of enzyme on feed intake in goats. Results
revealed that the supplementation of enzyme to TMR did not affect (P>0.05)
DMI. Further, the apparent digestibilities of DM, OM and CP were not affected
by enzyme supplementation.
Salem et al. (2012) conducted an experiment in sheep fed Atriplex
halimus (AH) foliages either fresh (AH-F) or sundried (AH-S) in the absence (-
ENZ) or presence (+ENZ) of 10 g/sheep/day of exogenous ZADO enzyme
preparation to assess the effects of sun drying and/or addition of an exogenous
enzyme (ENZ) preparation on intake and digestibility of nutrients. They
observed that enzyme addition to AH-S increased intake (P=0.001) as well as
OM and NDF digestibility (P=0.02). Further, N intake and N digestibility were
also higher (P=0.03) in AH-S sheep supplemented with ENZ.
In a complete random design, 12 mature male Ossimi sheep divided into
4 groups were fed 4 dietary treatments viz. Rice straw with grown barley
(RSGB) with neither ZAD (anaerobic enzyme) nor orange pulp (control, T1),
RSGB + ZAD (T2), RGSB + orange pulp (T3) and RSGB + ZAD + orange pulp
(T4). It was reported that the digestibility coefficients of DM, OM, CF, EE and
NFE were significantly higher in T4 than in T3, in T2 than in T1 (P<0.05). TDN
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and DCP values were higher in T4 than T3, in T2 than T1 (P<0.05) lowest being
in T1 (Gomaa et al., 2012).
Bhasker et al. (2013a) conducted an experiment on 12 male lambs by
randomly allotting to the two dietary groups viz. Sorghum stover based TMR
(R : C, 50 : 50) and TMR supplemented with EFE, to study the effect of EFE
supplementation on nutrient utilization. Results revealed that EFE
supplementation improved the digestibilities of OM, NFE, cell contents, NDF
and cellulose in sheep with no effect on CF and EE digestibilities. EFE
supplementation improved TDN and ME content while no effect was observed
on Ca and P balances. Similarly, in another study, 12 ram lambs were offered
maize stover based TMR (R : C, 50 : 50) with or without EFE. Results showed
that supplementation of EFE has no significant effect on DM, OM, CP, EE and
NFE digestibilities except for higher (P<0.05) CF digestibility. The
digestibilities of fibre fractions were comparable between the groups. Though
non-significant, a 4 % increase in DCP and TDN was observed upon EFE
supplementation (Bhasker et al., 2013b).
In another study, three fibrolytic enzyme extracts were evaluated using 20
Pelibuey lambs for their effects on feed digestibility, using diets containing
60% forage. The study indicated that there were no beneficial effects of
fibrolytic enzyme extracts on feed digestibility in lambs (Torres et al., 2013).
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2.2 EFFECT OF YEAST CULTURE SUPPLEMENTATION IN
RUMINANTS
There is scope for altering or manipulating rumen fermentation by yeast
culture supplementation and to increase the nutrient utilization and to improve
efficiency of production in farm animals (Kamra et al., 2002). The research
conducted using yeast culture in ruminants; the possible mode of action and the
effect of supplementation of yeast culture in the rations of ruminants on various
parameters are reviewed here.
2.2.1 MODE OF ACTION OF LIVE YEAST CULTURE
Effects and modes of action of yeast additives on rumen microbes have
been extensively studied over the last two decades. Several mechanisms have
been described, mostly from in vitro studies and also from studies with animal
models. Weidmeir and Arambel (1985) indicated that yeast supplementation
increased the concentrations of cellulose degrading bacteria in the rumen and
increased the relative concentration of acetate in the rumen. Addition of yeast
culture increased the number of anaerobic bacteria fivefold and total number of
cellulolytic bacteria two folds (Dawson et al., 1987).
Gunther (1989) reported that addition of live yeast cultures causes an
increase in energy supply to the micro organisms through slowly metabolizable
carbohydrates, decreased methane build up in the rumen, increase in the
efficiency of fermentation and increase in microbial protein synthesis which
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leads to improvement of feed conversion for better milk yield. Williams (1989)
indicated that addition of Yea-sacc to the rumen increases the number of
bacteria especially cellulolytic bacteria in the rumen possibly by associating in
H2 transfer, thereby allowing increased cellulolysis and reducing losses of H2
as methane. Erasmus (1991) suggested that yeast may have selective
stimulatory effect on specific rumen bacteria which result in a shift in the
protein synthesis and amino acid profiles.
Newbold (1991) reported that addition of yeast culture to the diets of
cows had reduced ammonia production, increased the flow of non ammonia
nitrogen to the duodenum, and increased fibre digestibility. Williams et al.
(1991) reported that presence of yeast culture in the rumen had an effect on
rumen stochiometry and increase the rate of forage degradation which might
result in increased forage intake and productivity. Karalazos et al. (1992)
suggested that addition of yeast to the ration increases the DM intake, the
relative concentration of the acetate produced in the rumen and ME content of
the diet through reduction in methane production.
Girard (1996) proposed the mode of action of the strain Yea-sacc1026
in
the rumen. Live yeast act by decreasing the lag time of specific ruminal
bacteria. The lag time is the time required by the bacteria to intiate growth and
enzyme production to digest substrate. He also reported that small peptides
from metabolically active yeast trigger exponential growth of ruminal bacteria.
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Beauchemin et al. (2006) proposed that increased bacterial numbers
within the rumen seems to be central to the action of yeast, driving both an
increased rate of fibre degradation in the rumen and an increased flow of
microbial protein from the rumen. Numerous studies (Srinivas Kumar et al.,
2013; Pinloche et al., 2013) documented positive effects of yeast, not only on
the rumen environment, but also on the improvement of microbial activities.
Pinloche et al. (2013) suggested that the beneficial action of live yeast is
due to either the potential oxygen scavenging activity of the yeast itself and/or
the stimulation of the general bacterial population. That process decreases the
inhibitory effect of oxygen on the rumen microflora (Newbold et al., 1996).
Yeast culture provides soluble growth factors (i.e., organic acids, B vitamins,
and amino acids) that stimulate growth of ruminal bacteria that utilize lactate
and digest cellulose (Callaway and Martin, 1997; Mao et al., 2013).
The positive effects of yeast can be attributed to the presence of mannans
and glucose polymers in yeast cell walls. MOS’s are capable of neutralizing
pathogenic bacteria, and they support β-glucans in the process of stimulating
defense mechanisms (Zabek et al., 2014).
2.2.2 EFFECT OF YEAST CULTURE SUPPLEMENTATION ON IN
VITRO DIGESTIBILITY
Malik and Singh (2009) evaluated twelve cultures of Saccharomyces
cerevisiae, added @ 106 cfu/conical flask and five cultures of Aspergillus
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oryzyae as probiotic supplement under in vitro condition using rumen liquor
from adult buffalo. Most of the strains of S. cerevisiae significantly increased
the IVDMD. Later, Fortina et al. (2011) conducted an in vitro study using 6
samples of rumen fluid to determine the DM, CP and NDF degradability of
total mixed ration at 0 and 48 h with the addition of 0 g (Y0) and 1 g (Y1) of
yeast culture. Results indicated that addition of 1 g YC to the rumen fluid
increased IVCPD and IVNDFD after 48 h of incubation but did not influence
IVDMD.
Nehra et al. (2013) conducted an in vitro study to determine the effect of
live yeast culture (YC) supplementation and optimum level of incorporation of
green gram straw in complete feed of goat. The study indicated that the YC
supplementation increased (P<0.01) the IVDMD and IVOMD of complete feed
irrespective of the level of green gram straw. Recently, Elghandour et al.
(2014) studied the effect of Saccharomyces cerevisiae on degradability of corn
stover, oat straw, sugarcane bagasse and sorghum straw and reported that the
direct addition or 72 hrs pre-incubation of Saccharomyces cerevisiae to
sorghum straw increased (P<0.05) DMD and NDFD.
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2.2.3 EFFECT OF YEAST CULTURE SUPPLEMENTAION ON
RUMEN FERMENTATION PATTERNS
2.2.3.1 Effect of Yeast culture supplementation in buffaloes
Gurpreet Singh et al. (2008) grouped 8 male buffalo calves into two
groups out of which I served as control while group II was supplemented with
Yea-sacc1026
at one bolus/animal/day for 21 days. They reported that rumen pH
in both the groups was decreased 3 h after feeding. However, the decline in
post-prandial pH was significantly less in the treatment group compared to the
control. Further, significantly higher values of TVFA, total N and NH3-N were
observed in the supplemented group compared with the control.
In a 4 x 4 LSD, four fistulated Murrah buffalo steers were fed a straw
based complete diet (12% CP) supplemented with 0, 0.1, 0.2 and 0.3% level of
thermo, acid, osmo and bile tolerant yeast (Saccharomyces cerevisiae OBV-9)
to study the effect on rumen fermentation pattern. Results revealed that the pH
of rumen liquor and TVFA concentration were similar among complete diets
whereas, ammonia nitrogen concentration was significantly (P<0.05) lower and
TCA-IPN concentration was significantly (P<0.05) higher on diets
supplemented with different levels of yeast as compared to the control (Bhima
et al., 2009).
Srinivas Kumar et al. (2011b) conducted an experiment on 6 graded
Murrah buffalo bulls by dividing into two groups of three animals each. All the
animals were maintained on 1.5 kg of concentrate mixture and hybrid napier
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(CO-1) fodder ad libitum while the experimental group was supplemented with
Levucell SC20 @ 0.5 g/animal/day. They reported that rumen pH, TVFA
concentration, NH3-N, total N, TCA-IPN, residual N as well as food and
protozoal N increased (P>0.05) with yeast culture supplementation in the diet
as compared to the control.
In 4 x 4 LSD, four graded Murrah buffalo bulls were randomly allotted
to four dietary treatments viz., maize stover based complete ration (T1), T1
supplemented with yeast culture (T2), jowar stover based complete rations (T3)
and T3 supplemented with yeast culture (T4). Results revealed that
supplementation of yeast culture in the complete rations increased (P<
0.01) the concentrations of mean pH, TVFA and N fractions irrespective of the
crop residue (Raj kiran et al., 2013).
2.2.3.2 Effect of Yeast culture supplementation in cattle
An experiment was conducted to study the effect of Saccharomyces
cerevisiae (SC) yeast culture (0 or 10 g/day) in Holstein steers (251 ± 2 kg) on
oat straw based diets. Results revealed that yeast culture supplementation had
no affect on rumen pH while TVFA concentration increased (P>0.05)
compared with the control (Plata et al., 1994).
Kamra et al. (2002) fed calves with roughage based diet supplemented
with or without yeast cell suspension and reported that pH (6.86 vs. 6.27)
increased (P<0.05), NH3-N concentration (20.1 vs. 23.9 mg/100 ml) decreased
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(P<0.05) and that of TVFA (10.7 vs. 11.2 mmol/100 ml) decreased (P>0.05) in
rumen liquor of the yeast group compared to the control.
Sixty multiparous HF cows in 4 groups were fed a total mixed ration
(Control) supplemented with yeast culture (Saccharomyces cerevisiae),
monensin, or both. Results revealed that YC had no effect on mean rumen pH
(5.69 vs. 5.79) and rumen ammonia N (6.35 vs. 7.63 mM) compared with the
control (Erasmus et al., 2005).
Moallem et al. (2009) studied the effects of live yeast (LY)
supplementation to dairy cows during the summer season. Forty two dairy
cows were fed either a control lactating diet or supplemented with 1 g of LY
(Saccharomyces cerevisiae, Biosaf, Lesaffre) per 4 kg of dry matter consumed.
Results indicated that ruminal ammonia concentrations after feeding were
greater in the control group than in the LY group (151.9 vs. 126.1 mg/l,
respectively). Further, the pH values in the rumen that were determined in a
companion trial using 4 fistulated cows tended to be higher in cows that were
supplemented with LY than in the control (6.67 vs. 6.54, respectively).
Ibrahim et al. (2012) investigated the effect of an abrupt or a gradual
introduction to pasture after calving and supplementation with live yeast
culture (YC) on ruminal pH and fermentation in early lactation. Rumen fluid
was harvested on d 8-10 and 22-24 post-partum (PP). Results revealed that
ruminal pH in the first measuring period (d 8-10 PP) were not affected by YC
supplementation while pH in the second measuring period (d 22-24 PP) was
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higher (P<0.01) in YC group than in control. The TVFA concentration was
greater (P<0.01) in YC group than in control while ammonia N concentration
was not affected by YC supplementation during both measuring periods.
In 3 x 3 LSD, three canulated lactating cows were fed a daily ration (24
kg/d) comprising of corn silage (61% of DM), concentrates (30% of DM),
dehydrated alfalfa (9% of DM) and a minerals and vitamins mix (1% of DM)
to study the effect of yeast (BIOSAF SC 47, Lesaffre Feed Additives, France;
0.5 or 5 g/d) as compared to a control (no additive). Results revealed that
supplementation of yeast increased the average pH (P<0.05), VFA (P<0.01)
and decreased the concentration of ammonia (P<0.05) (Pinloche et al., 2013).
2.2.3.3 Effect of Yeast culture supplementation in Sheep and Goat
Arcos-Garcia et al. (2000) conducted a 3 x 3 LSD trial using three
Suffolk ewes by allotting to three dietary treatments viz. control group (CG); 3
g/day of Yea sacc1026
(YS) and 1 g/day of Levucell (LC). They reported that
rumen pH was highest (P<0.05) in CG, and lowest (P<0.05) with YS than with
LC. Further, concentrations of rumen NH3-N were higher in YS and LC
compared to CG while the TVFA concentration was greater (P<0.05) with
yeast cultures (LC and YS) than in CG.
In 4 x 4 LSD, four fistulated rams were fed complete rations containing
GN haulms and concentrates in 60:40 or 70:30 ratios with or without yeast
culture to study the effect on rumen fermentation. Results revealed that
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ammonia nitrogen concentration was higher (P<0.05) in YCP supplemented
group while pH, concentration of total nitrogen and TVFA were comparable
among the treatments (Aasha Rekha et al., 2006).
Chandrashekar Patil et al. (2009) fed complete diet with or without yeast
culture in sheep and reported that the pH and NH3-N were found to be
significantly (P<0.01) lower and TVFA concentration was significantly
(P<0.01) higher in yeast fed group while total N, TCA perceptible N and
soluble N was similar among the groups.
Garg et al. (2009) conducted a trial using twelve male Magra lambs by
dividing into two equal groups and fed groundnut straw based complete feed
blocks with (YS) or without (YU) yeast (Saccharomyces cerevisiae) to study
the effect on rumen fermentation pattern. Yeast supplementation increased
rumen pH (P<0.01), TVFA concentration (P<0.01) and total N (P<0.05) while
the concentration of ammonia N decreased (P<0.05) with yeast culture
supplementation in the diet.
An experiment was conducted using 20 adult Nellore rams by dividing
into four groups and fed paddy straw based complete diets to study the effect
on rumen profile. The animals in the control group (T1) were fed chopped
paddy straw and concentrate mixture separately, while those in group 2 (T2)
complete ration in mash form, in group 3 (T3), T2 was supplemented with yeast
culture @ 0.1% level, and in group 4 (T4), T2 was subjected to expander-
extruder processing. Results indicated significantly (P<0.01) higher
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concentration of total-N, TCA-ppt-N, NH3-N and TVFA in groups T2, T3 and
T4 compared to T1 but differences among T2, T3 and T4 were not significant
(Chander Datt et al., 2011).
In 3 x 3 LSD, Kowalik et al. (2011) fed goats with a basal diet
comprising of hay (63%), barley meal (31%) and soybean meal (4%) (Control),
basal diet supplemented with live Saccharomyces cerevisiae (CNCM I-1077)
cells (R1) or their metabolites (R2). The additives were supplied @ 3 and 25
g/day, respectively. Results indicated that yeast metabolites decreased (P<0.01)
TVFA and increased rumen pH from 6.5 to 6.7 while live yeast had no effect
both on TVFA and rumen pH.
Sixty Ossimi male lambs were randomly assigned to six nutritional
groups and fed with two basal rations differing in roughage ratios (control 1 or
2) without or with supplementation of 0.1 or 0.2% dry yeast containing 108
cells of Saccharomyces cerevisiae per g to evaluate the effect of yeast culture
on rumen fermentation. Results indicated that yeast culture supplementation
increased (P<0.05) ruminal pH value and decreased (P<0.05) ammonia
concentration while there was no effect on TVFA concentration (Sawsan et al.,
2012).
Harikrishna et al. (2013) conducted a trial using 18 Nellore ram lambs
by randomly assigning to one of the three dietary treatments viz. diet with no
yeast (CON), diet with mesophilic yeast, 1 g/kg (MPY) and diet with
thermotolerant yeast, 1 g/kg (TPY) to evaluate the effect on rumen
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fermentation pattern. They reported that all the nitrogen fractions of rumen
fluid (except food and protozoal nitrogen), pH and TVFA were higher (P<0.01)
on TPY diet, compared to other two diets.
2.2.4 EFFECT OF YEAST CULTURE SUPPLEMENTATION ON
NUTRIENT DIGESTIBILITY AND NUTRITIVE VALUE
2.2.4.1 Effect of Yeast culture supplementation in Buffaloes
Mahender et al. (2005) fed 12 Murrah buffaloes in early lactation with
complete diets supplemented with yeast culture @ 0.1% (R1) or without yeast
culture (R2) and a conventional ration (R3) containing concentrate mixture,
maize fodder and sorghum straw ad lib and reported that the digestibility of
DM, OM, CF and NFE increased (P>0.05) while that of CP and EE increased
(P<0.05) significantly with supplementation of yeast in R1 when compared
with R2 and R3. Further, DCP (P<0.05) and TDN contents (P>0.05) were
higher in R1 compared to R2 and R3.
Srinivas Kumar et al. (2010) conducted a trial in 12 graded Murrah
buffalo bull calves by dividing into two equal groups to study the effect of
yeast culture supplementation on nutrient utilization. All the animals were
offered a conventional concentrate mixture @ 500 g/d and chopped Guinea
fodder ad libitum. In the yeast supplemented group, the concentrate mixture
was supplemented with Levucell SC20 at 0.25 g/animal/day. They reported that
the DMI (kg/100 kg BW) was lower (P>0.05) in the YC supplemented group
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compared to the control. The average digestibility coefficients of DM, EE and
NDF (P<0.05), OM, CP, CF, ADF and cellulose (P<0.01) increased
significantly with YC supplementation in the diet compared with the control. N
(P<0.01), Ca and P (P<0.05) balances (g/day) were significantly higher in the
YC supplemented group compared with the control. Further, % DCP (P>0.05)
and TDN (P<0.01) content increased with YC supplementation in the diet
compared with the control.
In another study, 12 graded Murrah buffaloes were divided into two
equal groups (Control and Treatment), to study the effect of yeast culture
supplementation on nutrient utilization. Animals in both the groups received a
basal diet comprising of roughages and concentrates to meet requirements
(ICAR, 1998). The animals in treatment group received yeast culture @ 0.5
g/animal/day. Results revealed that the DMI (kg/100 kg BW), DCP, TDN,
digestibility coefficients of gross nutrients and fibre fractions were not affected
by YC supplementation in the diet compared with the control (Srinivas Kumar
et al., 2011a).
Raj Kiran et al. (2014) reported that the digestibility coefficients of DM,
OM, CP, EE, CF, NDF, ADF, hemi-cellulose and cellulose increased (P<0.01)
with yeast culture (Saccharomyces cerevisiae) supplementation in complete
rations irrespective of the type of roughage.
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2.2.4.2 Effect of Yeast culture supplementation in Cattle
Kamra et al. (2002) fed 16 crossbred cattle calves in 2 groups given or
not a daily dose of 10 ml yeast cell suspension (YC) for 159 days and reported
that the digestibility of DM, OM, NDF and ADF decreased while that of CP and
EE increased with YC in the diet but the differences were not significant. In
another study, Reddy and Bhima (2003) fed 12 Deoni calves in two groups
with complete diets supplemented without or with lyophilized yeast culture at
0.1 % level and reported that the digestibility of DM, OM, CP, CF, EE, NFE, DCP
and TDN contents were significantly (P<0.05) higher on the diet containing
yeast culture compared with the control diet.
In a switch over design, Rajanna et al. (2005) divided 12 crossbred cows
into 2 groups (Control and Treatment) and fed a diet comprising of ragi straw
and concentrate mixture in 35: 65 ratio. Animals in treatment group received
10 g/day of yeast as a top dress to concentrate mixture. Results indicated that
the yeast culture supplementation had no influence on the whole tract
digestibility of DM, OM, CP, NDF and ADF.
Kishan Kumar and Ramana (2008) fed 12 Deoni calves in two groups
with a complete diet supplemented without (CD) or with yeast culture (YC) @
0.1% and reported that the digestibility of DM, OM, CP, EE, CF, NFE, NDF
and ADF were increased significantly (P<0.01) while DCP (6.3 vs. 5.8%) and
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TDN (70.1 vs. 67.4%) contents were increased (P>0.05) with YC in the diet
compared with the control.
Eighteen crossbred cows were randomly divided into three equal groups
of six each and were fed on concentrate mixture of same composition which
was supplemented with probiotic feed supplement @ 0 (Group I, Control), 10
(Group II) and 20 (Group III) g/day/cow, respectively. Results revealed that the
digestibility of DM, OM, CP, EE, CF and NFE and % DCP and TDN contents
were higher (P>0.05) in probiotic supplemented groups compared with the
control (Phondba et al., 2009).
Moallem et al. (2009) reported that live yeast (Saccharomyces cerevisiae,
Biosaf, Lesaffre, 1 g/ 4 kg DM consumed) supplementation to dairy cows
during the summer season had no effect on the apparent digestibility of DM,
OM, CP, NDF and ADF. Recently, Raval et al. (2013) reported that
supplementation of probiotics (containing Saccharomyces cerevisiae and
Lactobacillus sporogenes) in the diet of lactating Kankrej cows resulted in
increased (P<0.05) digestibility of DM and EE but had no effect on the
digestibility of OM, CP, CF and NFE.
2.2.4.3 Effect of Yeast culture supplementation in Sheep and Goats
Sixteen Nellore brown rams were divided into 4 groups of 4 animals
each and fed a complete ration with 60:40 roughage to concentrate ratio (CR-1)
supplemented with Lactobacillus acidophilus, 1 g (CR-2), Saccharomyces
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cerevisiae (Yea-sacc1026
), 10 g (CR-3) or L. acidophilus, 0.5g + Yea-sacc1026
, 5g
(CR-4). Results revealed that the CP digestibility in CR-1 was significantly
lower (P<0.01) than those in CR-2, CR-3 or CR-4. Improved fermentation
efficiency due to Yea-sacc1026
supplementation resulted in higher (P<0.01) CF
and cell wall constituents digestibility on CR-3 than on CR-1 (Nagamalleswara
Rao et al., 2001).
El-Ghani (2004) fed three Zaraibi bucks with concentrate mixture and
roughage (Control) supplemented with 3 g or 6 g of yeast culture (YC) and
reported a non significant increase (P>0.05) in the digestibility coefficients of
DM, OM and CP and significant increase in digestibility coefficients of CF
(P<0.01), EE (P<0.05) and NFE (P<0.01) for bucks fed YC than for controls.
Aasha Rekha et al. (2005) conducted studies to evaluate the groundnut
haulms (GNH) alone or with yeast culture (GNH-YC) as sole feed for Nellore
brown rams and reported that supplementation of yeast culture to GNH
improved the digestibility of nutrients considerably especially cellulose
(P<0.05). Further, they reported that the N and Ca retention were not affected
while P retention as per cent of absorbed P was improved (P<0.05) (67.1 vs.
75.6) when GNH were supplemented with yeast culture.
Eighteen Nellore ram lambs were fed on complete diets supplemented
with yeast culture @ 0.1% (R3) or without yeast culture (R2) and a
conventional ration (R1) containing concentrate mixture and chopped sorghum
straw ad libitum to study the effect on nutrient utilization. Results revealed that
111
yeast supplementation in the diet increased significantly (P<0.01) the
digestibility of DM, OM, CP, CF and NFE and increased DCP and TDN
contents (P>0.05) in ram lambs compared to un-supplemented diets. Further,
yeast supplementation in the diet increased significantly (P<0.01) N, Ca and P
balances in ram lambs compared to un-supplemented diets (Mahender et al.,
2006).
Titi et al. (2008) conducted an experiment to examine the effects of
inclusion of yeast culture (YC) to a diet based on barley grain and wheat straw
on digestibility of Awassi lambs. YC was added to the diet of treated group at
the level of 12.6 kg YC/tonne of diet. Results revealed that addition of YC had
no effect on apparent digestibility of DM, CP and NDF, but it increased
(P<0.05) digestibility of OM and ADF. Further, no differences were observed
in N intake, output or retention.
Chandrashekar Patil et al. (2009) fed complete diet with or without yeast
culture in sheep and reported that supplementation of yeast culture has no
significant effect on the digestibility of nutrients and forage fiber fractions
except cellulose, which increased significantly (P<0.01) in yeast fed group.
However, N, Ca and P balances, DCP and TDN intakes (g/d) were similar
among the groups.
Twelve male Magra lambs were fed groundnut straw based complete
feed blocks with (YS) or without (YU) yeast (Saccharomyces cerevisiae) to
study the effect on nutrient utilization. It was reported that the DMI (kg/100 kg
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BW), digestibility of DM, OM, gross nutrients and fibre fractions were
comparable between the two groups. The DCP and TDN contents were 10.53
and 51.82 per cent in YU group and 11.32 and 56.38 per cent in YS group,
respectively (Garg et al., 2009).
A complete diet was formulated and supplemented with three levels (D1 -
0 g/kg; D2 - 1 g/kg; D3 - 2 g/kg; D4 - 3 g/kg) of thermo-tolerant yeast
(Saccharomyces cerevisiae, OBV-9 @ 5Χ 108 cfu/g) to determine the best
level for sheep. Results revealed that the digestibility of dry matter, organic
matter, crude protein, crude fibre, EE, NFE and fibre fractions increased
significantly (P<0.01) on rations D2 to D4 over control, while the difference
among rations D2 to D4 was not significant (Harikrishna et al., 2012).
Sawsan et al. (2012) reported that addition of yeast culture to the basal
ration improved the digestion coefficients of DM, OM, CP and CF. Further, N
balance, TDN and DCP also increased (P<0.01) with addition of yeast culture
in the diet. Recently, Harikrishna et al. (2013) compared the effect of thermo-
tolerant yeast and mesophilic yeast compared to the control and reported that
the digestibility of DM, OM (P<0.05), CP, CF, NFE, NDF, ADF, hemi-
cellulose (P<0.05) and cellulose were higher (P<0.01) on thermo-tolerant yeast
diet as compared to the other two diets.
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2.3 EFFECT OF SUPPLEMENTATION OF EFE AND YEAST
CULTURE IN RUMINANT RATIONS
It was well established that supplementing nitrogen, readily fermentable
energy, minerals and monensin in licking blocks improved rumen fermentation
and nutrient utilization (Debasis and Singh, 2002). The research conducted
using yeast culture in combination with monensin or enzymes in ruminants;
and the effect of supplementation in the rations of ruminants on various
parameters are reviewed here.
2.3.1 EFFECT OF EFE AND YEAST CULTURE SUPPLEMENTATION
ON IN VITRO DIGESTIBILITY
An in vitro gas production experiment was conducted to determine the
effect of yeast culture (YC) and cellulolytic enzyme (CE) included in urea-
molasses-mineral (UMM) licking blocks on fibre degradation of wheat straw.
The treatment blocks include control (CON), typical block (B), yeast culture
block (YCB), cellulolytic enzyme block (CEB) and yeast culture plus
cellulolytic enzyme block (YCCEB). Results indicated that both DMD and
NDFD were significantly (P<0.05) enhanced by supplemental UMM licking
blocks, of which the greatest effect were found with YCB, followed by
YCCEB, CEB and B (Can et al., 2007).
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Tang et al. (2008) conducted an in vitro study to evaluate the effects of
yeast culture and fibrolytic enzyme preparation (containing cellulase and
xylanase) on fermentation characteristics of rice straw, wheat straw, maize
stover and maize stover silage. Four levels of yeast culture and fibrolytic
enzyme supplements (0, 2.5, 5 and 7.5 g/kg of straw DM, respectively) were
tested in a 4Χ4 factorial arrangement. They reported that supplementation of
yeast culture increased the IVDMD and IVOMD while fibrolytic enzyme
supplementation enhanced the IVDMD and IVOMD for 4 types of cereal
straws. Further, the study revealed significant interactions between EFE and
yeast on IVDMD and IVOMD of each type of straw.
2.3.2 EFFECT OF EFE AND YEAST CULTURE SUPPLEMENTATION
ON RUMEN FERMENTATION PATTERN
In 4 x 4 LSD, Garcia et al. (2000) conducted a trial using four Suffolk
sheep fitted with rumen cannula by allotting to four dietary treatments viz.
control, 1 g/d yeast culture (L), 25 mg/d of monensin (M) and a combination of
L and M, to study the effect of adding yeast culture, monensin or both on
rumen fermentation. Results indicated that the feed additives had no effect on
rumen pH and TVFA concentration.
In a CRD experiment, 60 multiparous HF cows were randomly allotted
to four dietary treatments viz. control diet (C), control plus 2550ppm (DM
basis) of yeast culture (YC), control plus 10 ppm (DM basis) of monensin (M)
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and control plus 2550ppm (DM basis) of YC plus 10 ppm (DM basis) of
monensin, to study the effect of feeding yeast culture, monensin or both on
rumen fermentation pattern. Results revealed that the mean postpartum rumen
pH and ammonia N concentrations were not affected by treatment, although pH
and ammonia N progressively decreased with either YC or M as the level of pH
or ammonia N in the cows increased (Erasmus et al., 2005).
Can et al. (2007) reported that supplementation of yeast culture,
cellulolytic enzymes or their combination included in licking blocks increased
(P<0.05) pH, ammonia N and TVFA concentration in the rumen.
Later, Bagheri et al. (2009) studied the effects of supplementation of
live yeast (SC), yeast cell-wall mannan-oligosaccharide (MOS) or both on
rumen fermentation pattern of cows during early lactation and reported that
either SC or MOS supplementation alone or in combination had no effect on
rumen pH, ammonia N and TVFA concentration.
Lopuszanska-Rusek and Bilik (2011) conducted a study by allotting 24
cows into 4 groups and fed 4 diets viz. control diet (C), control supplemented
with fibrolytic enzymes (E), control supplemented with yeast (D) and control
supplemented with E and D (ED). They reported that supplementation of diets
with enzyme or yeast preparation or both increased TVFA content and
individual fatty acid content in TVFA but the preparations had no effect on pH
and NH3-N concentration in rumen fluid pre and post-prandially.
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2.3.3 EFFECT OF EFE AND YEAST CULTURE SUPPLEMENTATION
ON NUTRIENT DIGESTIBILITY AND NUTRITIVE VALUE
Garcia et al. (2000) reported that addition of yeast culture, monensin or
both to the control diet in sheep had no effect on the digestibility of DM and
NDF. In another study, Malik and Bandla (2010) concluded that
supplementation of EFE along with probiotics improved (P<0.001)
significantly the OM, NDF and ADF digestibility.
Kung et al. (1997) fed the Holstein cows with a supplement containing
live yeast and enzymes twice daily with a diet of 50:50 (wt/wt) forage to
concentrate (DM basis). In the first lactation experiment, cows in midlactation
were offered a diet with corn silage as the primary forage source. The
supplement had no effect on pH, milk production, milk composition, or dry
matter intake in first lactation experiment.
Bagheri et al. (2009) studied the effects of supplementation of live yeast
(SC), yeast cell-wall mannan-oligosaccharide (MOS) or both on nutrient
digestibility of cows during early lactation and reported that SC
supplementation alone or in combination with MOS increased (P<0.05) the
apparent digestibility of DM and CP while there was no effect on NDF
digestibility. On the other hand, MOS supplementation alone had no effect on
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the apparent digestibility of nutrients. Further, supplementation of either SC or
MOS alone or in combination had no affect on DMI.
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CHAPTER – III
MATERIALS AND METHODS
In the present study, a total mixed ration with roughage: concentrate
ratio of 70: 30 was prepared incorporating groundnut haulms as a sole source
of roughage. The TMR as well as groundnut haulms were analyzed for
proximate principles and forage fibre constituents. Further, the total mixed
rations supplemented with exogenous fibrolytic enzyme (EFE) and/or live
yeast culture were evaluated through in vitro and in vivo methods for their
nutrient digestibility using graded Murrah buffalo bulls.
3.1 SOURCE OF EXOGENOUS FIBROLYTIC ENZYMES (EFE) AND
LIVE YEAST CULTURE
The exogenous fibrolytic enzyme (Fibrozyme) used in the present study
was procured from M/s Alltech Inc., Nicholasville, USA. The fibrozyme
(fermentation extracts of Aspergillus niger and Trichoderma viride containing
cellulases and hemicellulases; 100 IU as xylanase/g) was supplemented at the
rate of 2.5 g of enzyme powder /kg TMR (on DM basis). The live yeast culture
(Levucell SC 20 diluted) used in the present study was procured from
Lallemand, France. The live yeast culture (Saccharomyces cerevisiae 1- 1077)
containing 4 x 109 CFU / 10 g was supplemented at the rate of 10 g
/animal/day.
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3.2 PREPARATION OF TOTAL MIXED RATIONS
Total mixed ration (around 12% CP) containing roughage: concentrate
ratio of 70:30 was prepared using groundnut haulms as roughage source. The
ingredient composition of this complete ration is furnished in Table A.
3.3 IN VIVO STUDIES
3.3.1 Selection of animals
Four graded Murrah buffalo bulls, about 7 yrs of age with an average
body weight of 377.05 ± 43.36 kg each fitted with a permanent rumen fistula
were taken for the present study. All the animals were vaccinated against HS
and FMD and were dewormed one week prior to the start of the experiment.
3.3.2 Experimental design
The experimental TMRs were offered to the animals in a 4 x 4 Latin
square design allowing the switch over after every 28th
day.
3.3.3 Housing and management
The animals were housed in well ventilated conventional sheds
maintained in good hygienic condition and are stall fed throughout the
experimental period. Fresh, clean drinking water was provided to animals at
about 9.00AM and 3.00PM daily. Two days prior to the collection period, the
animals were shifted to the metabolism stalls for adaptation.
120
3.3.4 Feeding regimen
In 4 x 4 LSD, four graded Murrah buffalo bulls were randomly allotted
to four dietary treatments comprising of TMR supplemented with exogenous
fibrolytic enzyme (EFE) and/or live yeast culture as shown below. All the
animals were offered 6.5 kg each of respective dietary treatment (T1, T2, T3 and
T4) to meet the maintenance requirements (ICAR, 1998).
T1 – TMR containing R: C in 70:30 proportions
T2 – T1 supplemented with EFE @ 15 g/animal/day
T3 - T1 supplemented with live yeast culture @ 10 g/animal/day
T4 – T1 supplemented with EFE @ 15 g/animal/day plus live yeast
culture @ 10 g/animal/day
3.3.5 Weighing of animals
The animals were weighed weekly once during each period and at the
beginning and ending of the metabolism trial. Every time, weighing was done
on two consecutive days before offering feed and water and the average body
weight was calculated.
3.3.6 Metabolism trial
Each period of a Latin square consisted of a 21 days preliminary period
and 7 days collection period. The buffalo bulls were fed respective total mixed
rations at 9.00 AM and 3.00 PM all through the experimental period. On 20th
121
day of the preliminary period, the buffaloes were shifted to the metabolism
stalls for adaptation so as to reach their normal feed consumption. Daily feed
intake, feed refusals if any as well as faeces and urine voided were recorded
daily at 9.00 AM.
3.4 RUMEN FERMENTATION STUDIES
During the metabolism trial, at the end of each collection period, rumen
liquor was collected from rumen fistulated buffalo bulls just prior to feeding (0
h) and at 1, 2, 4, 6 and 8 h post feeding. On the days of rumen liquor collection,
feed was offered at 8 AM (before 0 h collection) and at 4 PM (after 8 h
collection) to avoid the effect of continuous feeding on concentration of rumen
metabolites. The animals were offered water one hour before start of collection
and after last collection to eliminate influence of water on nitrogen
concentration. The collected rumen liquor samples were strained through 4
layers of muslin cloth and resultant liquid was designated as strained rumen
liquor (SRL). About 100 ml of the SRL was drawn at each collection into a
clean sterile polythene bottle. pH of rumen liquor was determined immediately
by using digital pH meter.
The ammonia nitrogen in SRL was determined immediately after
collection. The remaining SRL was preserved after adding 1 ml of saturated
mercuric chloride to stop the microbial activity. All the samples were stored in
polyethylene bottles and preserved at sub-zero temperature for further analysis.
122
3.5 IN VITRO STUDIES
The four dietary treatments comprising of TMR supplemented with
exogenous fibrolytic enzyme (EFE) and/or live yeast culture were evaluated for
in vitro DM, CP, NDF and ADF digestibility (Tilley and Terry, 1963) with
strained rumen liquor (SRL) collected from rumen fistulated graded Murrah
buffalo bulls maintained on a conventional diet.
3.6 COLLECTION OF SAMPLES
3.6.1. Feed and feed residue
Representative samples of feed offered and feed residue if any during
each collection period were collected and pooled for 7 days and were latter
ground separately in a laboratory Wiley mill through 2 mm screen and
preserved in air tight bottles for proximate and fibre analysis.
3.6.2 Faeces
Twenty four hour collection of faeces was recorded on every day
morning at 9.00 AM for seven days. The faeces were weighed, mixed
thoroughly and representative sample (2%) was taken in polythene bag
separately for each animal and stored at -10ºC in a deep freeze.
After completion of each trial, pooled faecal samples were thawed to
room temperature mixed properly and fresh samples were taken for nitrogen
(8-10g) and DM (50-60g) analysis. For further analysis, faeces was dried at
60ºC and ground to pass through 1 mm screen and preserved in air tight bottles.
123
3.6.3 Urine
Twenty four hour collection of urine was recorded on every day
morning at 9.00 AM for seven days. The urine voided by each animal was
measured, mixed thoroughly and representative sample (2%) was taken in glass
bottle for each animal and stored at 4ºC in a refrigerator after addition of few
drops of concentrated sulphuric acid.
3.7 ANALYTICAL METHODS
3.7.1 Proximate Analysis
Samples of groundnut haulms, TMR and faeces were analyzed for
proximate constituents and urine for N according to AOAC (2007) methods.
The crude protein (N x 6.25) was estimated with fresh faeces samples. Nitrogen
analysis was done by using Turbotherm and Vapodest (Gerhardt, Germany)
analyzer. The crude protein (N x 6.25) of feeds and leftovers were also
estimated with fresh material.
3.7.2 Analysis of cell-wall constituents
Cell-wall constituents viz., neutral detergent fibre (NDF), acid detergent
fibre (ADF), cellulose, acid detergent lignin (ADL), and silica were determined
for feeds and faeces by using the methods described by Van Soest et al. (1991).
Hemi-cellulose was calculated as the difference between NDF and ADF.
3.7.3 Analysis of minerals
124
Calcium and Phosphorus in feeds and faeces were determined according
to methods described by Talapatra et al., (1940). Calcium and Phosphorus in
urine samples were determined by Ferro and Ham (1957) and Fiske and Subba
Row (1925), respectively.
3.7.4 Rumen metabolic profile studies
Rumen metabolic profile studies in terms of rumen pH, ammonia
nitrogen (NH3-N) and total volatile fatty acids (TVFA) concentration were
determined in strained rumen liquor (SRL). The pH of the rumen liquor was
measured immediately after collection of rumen liquor using digital pH meter
and ammonia nitrogen was determined by micro-diffusion method of Conway
(1957) using mixed indicator (Livingston et al., 1964). The TVFA
concentration of SRL was determined by using the procedure of Barnett and
Reid (1957). The total nitrogen (Micro-kjeldahl), TCA-insoluble protein
nitrogen (Cline et al., 1958), residual nitrogen, food and protozoal nitrogen
(Singh et al., 1968) were also determined.
3.8 STATISTICAL ANALYSIS
The data was analyzed statistically (Snedecor and Cochran, 1994) and
tested for significance using Latin Square Design and Duncan’s multiple range
test (Duncan, 1955) using SPSS 17.0 version.
125
Table A. Ingredient composition of the total mixed ration fed to graded
Murrah buffalo bulls during the metabolism trial
Ingredient Total Mixed Ration
Ground nut haulms 70.0
Maize grain 8.1
De Oiled Rice Bran 10.5
Cotton seed cake 7.5
Sunflower cake 3.0
Mineral mixture 0.6
Salt 0.3
Total 100.0
126
CHAPTER - IV
RESULTS
Four dietary treatments comprising of total mixed ration (TMR) with a
roughage: concentrate ratio of 70:30 supplemented with exogenous fibrolytic
enzyme (EFE) and/or live yeast culture are evaluated for their effect on nutrient
digestibility, balance of minerals, nutritive value and rumen fermentation
pattern using four rumen fistulated graded Murrah buffalo bulls. Further, the
TMRs supplemented with EFE and/or live Yeast cultures are also evaluated
through in vitro methods to study the degradability of DM, CP, NDF and ADF.
4.1 EVALUATION OF TOTAL MIXED RATIONS
4.1.1 Chemical composition and cell-wall constituents of Groundnut
haulms
The chemical composition of groundnut haulms used in the present
study is presented in Table 1. The per cent DM, OM, TA, CP, EE, CF and NFE
of groundnut haulms were 88.36, 90.90, 9.10, 9.3, 1.56, 37.80 and 42.24,
respectively. The per cent NDF, ADF, ADL, hemi-cellulose, cellulose and
silica content of groundnut haulms were 56.42, 48.89, 10.49, 7.53, 36.94 and
1.98, respectively. The per cent calcium and phosphorous content of groundnut
haulms were 1.07 and 0.20, respectively.
127
Table 1: Chemical composition (on % DM basis) of Groundnut haulms
Nutrient Groundnut haulms
Dry matter 88.36
Organic matter 90.90
Total ash 9.10
Crude protein 9.30
Ether extract 1.56
Crude fibre 37.80
Nitrogen free extract 42.24
Neutral Detergent Fibre 56.42
Acid Detergent Fibre 48.89
Acid Detergent Lignin 10.49
Hemi-cellulose 7.53
Cellulose 36.94
Silica 1.98
Calcium (%) 1.07
Phosphorus (%) 0.20
128
4.1.2 Chemical composition and cell-wall constituents of total mixed
rations
The chemical composition of groundnut haulms based total mixed ration
(TMR) with a roughage: concentrate ratio of 70:30 which was fed to buffalo
bulls during the metabolism trial is presented in Table 2. The per cent DM,
OM, TA, CP, EE, CF and NFE content were 92.82, 90.88, 9.12, 12.25, 1.54,
27.62 and 49.47, respectively.
The cell wall constituents of groundnut haulms based TMR fed to
buffalo bulls during the metabolism trial is presented in Table 2. The per cent
NDF, ADF, ADL, hemi-cellulose and cellulose were 53.22, 43.24, 9.77, 9.98
and 31.59, respectively. The per cent Ca and P contents were 1.03 and 0.49,
respectively.
4.2 IN VITRO STUDIES
The in vitro digestibility of dry matter (DM), crude protein (CP), neutral
detergent fibre (NDF) and acid detergent fibre (ADF) of total mixed ration
supplemented with exogenous fibrolytic enzyme and/or yeast culture was
studied using rumen liquor collected from buffalo bulls maintained on a
standard basal diet and is presented in table 3.
129
Table 2: Chemical composition (on % DM basis) of the total mixed ration
fed to buffalo bulls during the study
Nutrient TMR
Dry matter 92.82
Organic matter 90.88
Total ash 9.12
Crude protein 12.25
Ether extract 1.54
Crude fibre 27.62
Nitrogen free extract 49.47
Neutral Detergent Fibre 53.22
Acid Detergent Fibre 43.24
Acid Detergent Lignin 9.77
Hemi-cellulose 9.98
Cellulose 31.59
Silica 2.24
Calcium (%) 1.03
Phosphorus (%) 0.49
130
4.2.1 In vitro digestibility of DM in total mixed rations
The in vitro DM digestibility (%) was 50.97, 55.69, 56.63 and 57.02 in
T1, T2, T3 and T4, respectively. The % IVDMD was higher (P<0.05) in T4 and
lower (P<0.05) in T1
when compared to other treatments. However, no
significant (P>0.05) differences were observed among T2, T3 and T4 groups.
4.2.2 In vitro digestibility of CP in total mixed rations
The in vitro CP digestibility (%) was 53.98, 58.43, 59.64 and 60.57 in
T1, T2, T3 and T4, respectively. The % IVCPD was higher (P<0.05) in T4 and
lower (P<0.05) in T1 as compared to other treatments. However, no significant
(P>0.05) differences were observed among T2, T3 and T4 groups.
4.2.3 In vitro digestibility of NDF in total mixed rations
The in vitro NDF digestibility (%) was 49.15, 54.41, 55.28 and 57.28 in
T1, T2, T3 and T4, respectively. The % IVNDFD was higher (P<0.05) in T4 and
lower (P<0.05) in T1 as compared to other treatments. However, no significant
(P>0.05) differences were observed among T2, T3 and T4 groups.
4.2.4 In vitro digestibility of ADF in total mixed rations
131
The in vitro ADF digestibility (%) was 46.42, 51.94, 53.56 and 55.00 in
T1, T2, T3 and T4, respectively. The % IVADFD was higher (P<0.05) in T4 and
lower (P<0.05) in T1 as compared to other treatments. However, no significant
(P>0.05) differences were observed among T2, T3 and T4 groups.
132
Table 3: In vitro digestibility (%) of Total Mixed Ration supplemented with EFE and/or live yeast culture
Nutrient digestibility (%) T1 T2 T3 T4
IVDMD* 50.97a ± 0.50 55.69
b ± 0.51 56.63
b ± 0.83 57.02
b ± 1.27
IVCPD* 53.98a ± 0.50 58.43
b ± 1.00 59.64
b ± 0.95 60.57
b ± 0.92
IVNDFD* 49.15a ± 0.88 54.41
b ± 1.37 55.28
b ± 1.39 57.28
b ± 0.74
IVADFD* 46.42a ± 0.72 51.94
b ± 0.59 53.56
b ± 0.72 55.00
b ± 2.48
ab values in the rows bearing different superscripts differ significantly
*(P<0.05)
133
4.3 RUMEN FERMENTATION STUDIES
The rumen fermentation pattern in respect of pH, total volatile fatty acids
(TVFA) and nitrogen fractions in strained rumen liquor (SRL) of buffalo bulls fed
total mixed ration supplemented with exogenous fibrolytic enzyme and/or live yeast
culture is presented in Tables 4 to 10.
4.3.1 Rumen pH
The mean pH values of rumen fluid of buffalo bulls as affected by feeding
total mixed ration supplemented with exogenous fibrolytic enzyme and/or live yeast
culture with time of sampling are presented in Table 4. The mean values of pH were
6.81 ± 0.04, 6.91 ± 0.04, 6.95 ± 0.07 and 7.01 ± 0.02 for T1, T2, T3 and T4,
respectively. The mean pH values were significantly higher (P<0.01) in T4 and
lower (P<0.01) in T1. However, no significant differences were observed between T1
and T2, T2 and T3, and T3 and T4.
134
There was a significant difference (P<0.01) due to time of sampling with
respect to pH concentration. pH of rumen liquor showed a decreasing trend up to 4 h
post feeding in all the buffalo bulls irrespective of the treatment prior to attaining
normal levels. Diet x hour interaction was not significantly different for rumen pH.
135
Table 4: Rumen pH in buffalo bulls fed total mixed ration supplemented with exogenous fibrolytic enzymes and/or
live yeast culture
Hour 0 2 4 6 8 Mean ± SE**
T1 6.95 ± 0.03 6.72 ± 0.04 6.57 ± 0.03 6.82 ± 0.04 6.98 ± 0.07 6.81A ± 0.04
T2 7.03 ± 0.05 6.83 ± 0.05 6.74 ± 0.04 6.90 ± 0.02 7.07 ± 0.03 6.91AB
± 0.04
T3 7.08 ± 0.06 6.86 ± 0.08 6.76 ± 0.08 6.93 ± 0.07 7.11 ± 0.06 6.95
BC ± 0.07
T4 7.12 ± 0.03 6.94 ± 0.01 6.79 ± 0.01 7.04 ± 0.05 7.14 ± 0.04 7.01C ± 0.02
Mean ± SE** 6.88d ± 0.04
6.78
b ± 0.04
6.74
a ± 0.04
6.78
c ± 0.04
6.86
d ± 0.04
ABC Values in the columns bearing different superscripts differ significantly.
abcd Values in the rows bearing different superscripts differ significantly
**P<0.01 (n = 4)
136
4.3.2 Total volatile fatty acids (TVFA)
The total volatile fatty acid (TVFA) concentration in SRL of buffalo bulls fed
total mixed ration supplemented with exogenous fibrolytic enzyme and/or live yeast
culture with time of sampling are presented in Table 5. The mean TVFA
concentration in SRL of buffalo bulls were 81.56 ± 0.21, 86.51 ± 0.08, 87.86 ± 0.32
and 89.46 ± 0.10 meq/L in T1, T2, T3 and T4, respectively. The mean TVFA
concentration (meq/L) was higher (P<0.01) in T4 and lower (P<0.01) in T1 as
compared to other treatments.
Time after feeding linearly increased (P<0.01) the TVFA concentration up to
4 h post feeding beyond which there was a decline in its concentration. Significant
(P<0.05) diet x hour interaction was observed.
4.3.3 Ammonia nitrogen
The ammonia nitrogen concentration in SRL of buffalo bulls fed total mixed
ration supplemented with exogenous fibrolytic enzyme and/or live yeast culture is
presented in Table 6. The mean ammonia nitrogen concentration in SRL of buffalo
bulls was 10.12 ± 0.12, 10.97 ± 0.08, 11.15 ± 0.08 and 11.33 ± 0.05 mg/100 ml SRL
in T1, T2, T3 and T4, respectively. The mean ammonia nitrogen concentration was
higher (P<0.01) in T4 and lower in T1 when compared to other treatments. However,
no significant difference was observed between T2 and T3 and T3 and T4.
137
Table 5: TVFA concentration (meq/L of SRL) in buffalo bulls fed total mixed ration supplemented with exogenous
fibrolytic enzymes and/or live yeast culture
ABCDValues in the columns bearing different superscripts differ significantly.
abcdValues in the rows bearing different superscripts differ significantly
**
P<0.01 (n = 4)
Hour 0 2 4 6 8 Mean ± SE**
T1 67.06 ± 0.66 87.13 ± 0.48 94 ± 0.98 85.31 ± 0.98 74.31 ± 0.77 81.56A ± 0.21
T2 69.25 ± 0.55 90.75 ± 0.85 100 ± 0.53 90.81 ± 0.37 80.81 ± 0.80 86.51B ± 0.08
T3 70.94 ± 0.64 92.25 ± 0.85 102.56 ± 0.79 91.94 ± 0.65 81.63 ± 0.69 87.86C ± 0.32
T4 71.75 ± 0.71 93.94 ± 0.68 104.06 ± 0.80 93.94 ± 0.50 83.63 ± 0.55 89.46D ± 0.10
Mean ± SE** 69.75a ± 1.04
91.02
c ± 1.45
100.39
d ± 2.22
90.50
c ± 1.85
80.09
b ± 2.02
138
Rumen ammonia nitrogen concentration increased linearly (P<0.01) up to 4 h
post feeding beyond which there was a decline in its concentration. No Diet x hour
interaction was observed for ammonia nitrogen concentration.
4.3.4 Total nitrogen
The concentration of total nitrogen in SRL of buffalo bulls as affected by
feeding total mixed ration supplemented with exogenous fibrolytic enzyme and/or
live yeast culture is presented in Table 7. The mean values for total nitrogen
concentration were 75.75 ± 0.33, 81.40 ± 0.12, 83.25 ± 0.50 and 85.00 ± 0.47
mg/100 ml SRL in T1, T2, T3 and T4, respectively. The mean total nitrogen
concentration in SRL of buffalo bulls was higher (P<0.01) in T4 and lower (P<0.01)
in T1 when compared to other treatments.
The total nitrogen concentration increased (P<0.01) linearly up to 4 h post
feeding in all the buffalo bulls irrespective of the treatment beyond which there was
a decline in its concentration. No Diet x hour interaction was observed for total
nitrogen concentration in the SRL.
4.3.5 TCA insoluble protein nitrogen
The TCA insoluble protein nitrogen concentration in SRL of buffalo bulls fed
total mixed ration supplemented with exogenous fibrolytic enzyme and/or live yeast
culture is presented in Table 8. The mean values of TCA insoluble protein
139
Table 6: Ammonia nitrogen concentration (mg/100 ml SRL) in buffalo bulls fed total mixed ration supplemented
with exogenous fibrolytic enzymes and/or live yeast culture
Hour 0 2 4 6 8 Mean ± SE**
T1 8.01 ± 0.24 11.92 ± 0.20 12.93 ± 0.14 9.85 ± 0.19 7.91 ± 0.38 10.12A ± 0.12
T2 9.16 ± 0.06 12.58 ± 0.33 13.65 ± 0.28 10.68 ± 0.17 8.78 ± 0.09 10.97B ± 0.08
T3 9.26 ± 0.09 12.81 ± 0.34 13.91 ± 0.19 10.80 ± 0.28 8.95 ± 0.05 11.15BC
± 0.08
T4 9.44 ± 0.15 13.09 ± 0.23 14.08 ± 0.13 10.93 ± 0.18 9.11 ± 0.13 11.33C
± 0.05
Mean ± SE** 8.97a ± 0.32
12.60
c ± 0.25
13.64
d ± 0.25
10.56
b ± 0.24
8.69
a ± 0.27
ABCValues in the columns bearing different superscripts differ significantly.
abcd Values in the rows bearing different superscripts differ significantly
**P<0.01 (n = 4)
140
Table 7: Total nitrogen concentration (mg/100 ml SRL) in buffalo bulls fed total mixed ration supplemented with
exogenous fibrolytic enzymes and/or live yeast culture
Hour 0 2 4 6 8 Mean ± SE**
T1 62.75 ± 0.48 83.25 ± 0.85 94.50 ± 0.65 74.50 ± 0.65 63.75 ± 0.48 75.75A ± 0.33
T2 69.25 ± 0.63 88.50 ± 0.65 98.50 ± 0.65 80.25 ± 0.63 70.50 ± 0.87 81.40B ± 0.12
T3 71.00 ± 0.41 91.00 ± 0.41 100.75 ± 0.85 82.50 ± 0.65 71.00 ± 1.08 83.25C ± 0.50
T4 71.50 ± 0.65 93.00 ± 0.41 103 ± 0.91 84.25 ± 0.85 73.25 ± 0.85 85.00D ± 0.47
Mean ± SE** 68.63a ± 2.02
88.94
d ± 2.11
99.19
e ± 1.81
80.38
c ± 2.12
69.63
b ± 2.05
ABCDValues in the columns bearing different superscripts differ significantly.
abcdeValues in the rows bearing different superscripts differ significantly
**
P<0.01 (n = 4)
141
nitrogen concentration were 25.40 ± 0.35, 27.03 ± 0.49, 27.55 ± 0.32 and 28.33 ±
0.19 mg/100ml SRL in T1, T2, T3 and T4, respectively. The mean TCA insoluble
protein nitrogen concentration was significantly higher (P<0.01) in T4 and lower
(P<0.01) in T1 compared to other treatments while there was no significant
difference between T2 and T3 and T3 and T4.
Time after feeding significantly (P<0.01) affected the TCA insoluble protein
nitrogen concentration in SRL of buffalo bulls. The TCA insoluble protein nitrogen
reached peak concentration at 4 h post feeding beyond which there was a decline.
Diet x hour interaction was not significant (P>0.05) for TCA insoluble protein
nitrogen concentration in SRL.
4.3.6 Residual nitrogen
The values representing residual nitrogen concentration in SRL of buffalo
bulls fed total mixed ration supplemented with exogenous fibrolytic enzyme and/or
live yeast culture is presented in Table 9. The mean residual nitrogen concentration
(mg/100ml SRL) was higher (P<0.01) in T4 and lower (P<0.01) in T1 and the mean
values were 22.41 ± 0.34, 24.60 ± 0.22, 25.24 ± 0.12 and 25.76 ± 0.27 in T1, T2, T3
and T4, respectively. However, no significant difference was observed between T3
and T4.
142
143
Table 8: TCA insoluble protein nitrogen (mg/100 ml SRL) in buffalo bulls fed total mixed ration supplemented with
exogenous fibrolytic enzymes and/or live yeast culture
Hour 0 2 4 6 8 Mean ± SE**
T1 21.50 ± 0.35 29.50 ± 0.20 34.75 ± 0.66 24.00 ± 0.91 17.25 ± 0.52 25.40A ± 0.35
T2 23.88 ± 1.13 30.88 ± 0.83 35.63 ± 0.52 25.63 ± 0.88 19.13 ± 0.31 27.03B ± 0.49
T3 24.38 ± 0.77 31.63 ± 0.55 36.38 ± 0.52 26.13 ± 0.90 19.25 ± 0.52 27.55BC
± 0.32
T4 25.00 ± 0.84 32.13 ± 0.83 37.13 ± 0.55 27.25 ± 0.66 20.13 ± 0.43 28.33C ± 0.19
Mean ± SE** 23.69b ± 0.76
31.03
d ± 0.57
35.97
e ± 0.51
25.75
c ± 0.68
18.94
a ± 0.60
ABCValues in the columns bearing different superscripts differ significantly.
abcdeValues in the rows bearing different superscripts differ significantly
**P<0.01 (n = 4)
144
Time after feeding significantly (P<0.01) affected the residual nitrogen
concentration in SRL of buffalo bulls. Residual nitrogen concentration in the SRL
increased (P<0.01) linearly up to 4 h post feeding after which the values decreased.
No significant diet x hour interaction was observed.
4.3.7. Food and Protozoal nitrogen
The concentration of food and protozoal nitrogen in SRL of buffalo bulls fed
mixed ration supplemented with exogenous fibrolytic enzyme and/or live yeast
culture is presented in Table 10. The mean values of food and protozoal nitrogen
(mg/100ml SRL) in SRL of buffalo bulls were 17.81 ± 0.38, 18.81 ± 0.26, 19.31 ±
0.15 and 19.59 ± 0.69 in T1, T2, T3 and T4, respectively.
Diet (P<0.05) and time after feeding (P<0.01) significantly affected the food
and protozoal nitrogen concentration in SRL of buffalo bulls while diet x hour
interaction was non-significant. The mean food and protozoal nitrogen concentration
(mg/100ml SRL) was higher (P<0.05) in T4 and lower (P<0.05) in T1 compared to
other treatments. But, no significant differences were observed between T1 and T2,
T1 and T3, T2 and T3 and T3 and T4. Peak concentration of food and protozoal
nitrogen was observed 4 h post feeding beyond which there was a decline in its
concentration irrespective of the treatment.
145
Table 9: Residual nitrogen (mg/100 ml SRL) in buffalo bulls fed total mixed ration supplemented with exogenous
fibrolytic enzymes and/or live yeast culture
Hour 0 2 4 6 8 Mean ± SE**
T1 20.74 ± 0.56 23.64 ± 0.22 25.70 ± 0.89 21.09 ± 0.63 20.90 ± 0.47 22.41A ± 0.34
T2 22.78 ± 0.36 25.05 ± 0.67 27.35 ± 0.19 23.51 ± 0.21 24.29 ± 0.88 24.60B ± 0.22
T3 23.43 ± 0.19 26.31 ± 0.45 28.46 ± 0.48 24.64 ± 0.35 23.36 ± 0.31 25.24C ± 0.12
T4 23.19 ± 0.07 27.35 ± 0.45 29.68 ± 0.48 24.76 ± 0.59 23.83 ± 0.47 25.76C ± 0.27
Mean ± SE** 22.53a ± 0.61
25.59
c ± 0.80
27.80
d ± 0.84
23.50
b ± 0.85
23.09
ab ± 0.76
ABCValues in the columns bearing different superscripts differ significantly.
abcdValues in the rows bearing different superscripts differ significantly
**P<0.01 (n = 4)
146
Table 10: Food and protozoal nitrogen (mg/100 ml SRL) in buffalo bulls fed total mixed ration supplemented with
exogenous fibrolytic enzymes and/or live yeast culture
Hour 0 2 4 6 8 Mean ± SE*
T1 12.50 ± 0.74 18.19 ± 0.78 21.13 ± 1.01 19.56 ± 1.68 17.69 ± 0.90 17.81A ± 0.38
T2 13.44 ± 1.46 20.00 ± 1.47 21.88 ± 0.77 20.44 ± 1.33 18.31 ± 1.37 18.81AB
± 0.26
T3 13.94 ± 0.53 20.25 ± 0.68 22.00 ± 0.14 20.94 ± 1.43 19.44 ± 1.30 19.31AB
± 0.15
T4 13.88 ± 1.17 20.44 ± 1.24 22.13 ± 0.75 21.31 ± 1.46 20.19 ± 1.18 19.59B ± 0.69
Mean ± SE** 13.44a ± 0.33
19.72
b ± 0.52
21.78
c ± 0.22
20.56
bc ± 0.38
18.91
b ± 0.56
ABValues in the columns bearing different superscripts differ significantly.
abc Values in the rows bearing different superscripts differ significantly
*P<0.05
**P<0.01 (n = 4)
147
4.4 METABOLISM STUDIES IN MURRAH BUFFALO BULLS
4.4.1 Apparent nutrient digestibility coefficients
The apparent nutrient digestibility coefficients in buffalo bulls fed total
mixed rations are presented in Table 11. The digestibility coefficients of DM ranged
from 55.21 to 58.16, OM from 59.88 to 62.62, CP from 60.31 to 62.51, EE from
66.79 to 69.61, CF from 48.52 to 51.86 and NFE from 66.70 to 69.15 in buffalo
bulls fed total mixed ration supplemented with exogenous fibrolytic enzyme and/or
live yeast culture. The digestibility coefficients of DM, OM, CP, EE, CF and NFE
were numerically higher in T4 followed by T3 and T2 as compared to T1 but the
differences between treatments were not statistically significant (P>0.05).
The per cent digestibilities of cell wall constituents in buffalo bulls fed total
mixed rations are presented in Table 10. The digestibility coefficients of NDF
ranged from 50.60 to 55.19, ADF from 47.18 to 52.50, hemi-cellulose from 63.85 to
66.85 and cellulose from 59.16 to 63.78. The digestibility coefficients of NDF,
ADF, hemi-cellulose and cellulose were higher in T4 when compared to other
treatments. However, no significant (P>0.05) differences were observed between the
treatments.
148
Table 11: Apparent digestibility (%) of nutrients in buffalo bulls fed total mixed ration supplemented with
exogenous fibrolytic enzymes and/or live yeast culture
Nutrient T1 T2 T3 T4
Dry matterNS
55.21 ± 0.98 57.08
± 1.03 57.84
± 1.85 58.16
± 1.11
Organic matterNS
59.88 ± 1.16 61.94
± 0.83 62.30
± 1.47 62.60
± 1.10
Crude proteinNS
60.31 ± 1.29 62.08
± 2.13 62.43
± 1.75 62.51
± 1.48
Ether extractNS
66.79 ± 2.50 68.43
± 2.43 69.16
± 2.63 69.61
± 2.23
Crude fibreNS
48.52 ± 2.53 51.29
± 1.49 51.58
± 2.44 51.86
± 1.53
Nitrogen free extractNS
66.70± 0.69 68.60 ± 1.04 68.74
± 1.67 69.15
± 2.29
Neutral detergent
fibreNS
50.60 ± 1.92 53.50
± 1.36 54.51
± 1.29 55.19
± 1.62
Acid detergent fibreNS
47.18 ± 2.08 50.61
± 1.96 51.76
± 1.34 52.50
± 1.52
Hemi celluloseNS
63.85 ± 5.07 66.01
± 3.08 66.42
± 3.38 66.85
± 2.49
CelluloseNS
59.16 ± 0.40 62.55
± 1.55 63.17
± 0.77 63.78
± 1.00
NS – Non Significant
149
4.4.2 Nitrogen balance
Daily nitrogen intake, outgo and retention in buffalo bulls fed total mixed
ration supplemented with exogenous fibrolytic enzyme and/or live yeast culture are
presented in Table 12. The N excretion through faeces, urine and total N excretion
(g/d) were higher in T1 and lower in T4 but differences between treatments were not
significant (P>0.05). All the animals were in positive balance for nitrogen and the
average nitrogen retentions (g/d) were 59.44, 62.14, 64.77 and 65.16 in T1, T2, T3
and T4, respectively. The N retention expressed as either g/d, as per cent of intake or
as per cent absorbed were higher in T4 and lower in T1 when compared to other
treatments. However, no significant (P>0.05) differences between treatments were
observed.
4.4.3 Calcium balance
Calcium intake, outgo and retention in buffalo bulls fed total mixed ration
supplemented with exogenous fibrolytic enzyme and/or live yeast culture are
presented in Table 13. The calcium excretion through faeces and total calcium
excretion (g/d) was higher in T1 while calcium excretion through urine was higher in
T2. However, no significant differences (P>0.05) were observed between the
treatments. All the animals were in positive calcium balance. The calcium retentions
(g/day) were 19.95, 20.76, 22.82 and 24.54 in T1 T2, T3 and T4, respectively. The
150
Table 12: Nitrogen utilization in buffalo bulls fed total mixed ration supplemented with exogenous fibrolytic
enzymes and/or live yeast culture
T1 T2 T3 T4
Nitrogen intake, g/d 127.16 127.16 127.16 127.16
Nitrogen outgo, g/d
FaecesNS
50.55 ± 1.64 48.30
± 2.71 46.10
± 2.22 47.74
± 1.88
UrineNS
17.17 ± 1.67 16.72
± 1.43 16.29
± 0.54 14.25
± 4.26
TotalNS
67.72 ± 2.50 65.02
± 2.06 62.38
± 2.03 62.00
± 5.00
Nitrogen retention
Retention, g/dNS
59.44 ± 2.50 62.14
± 2.06 64.77 ± 2.03 65.16
± 5.00
% intakeNS
46.74 ± 1.97 48.87
± 1.62 50.94
± 1.59 51.25
± 3.93
% absorbedNS
77.53 ± 2.37 78.85
± 1.44 79.89
± 0.63 81.96
± 5.33
NS – Non Significant
151
Table 13: Calcium utilization in buffalo bulls fed total mixed ration supplemented with exogenous fibrolytic enzymes
and/or live yeast culture
T1 T2 T3 T4
Calcium intake, g/d 62.12 62.12 62.12 62.12
Calcium outgo, g/d
FaecesNS
37.81 ± 2.88 36.59
± 0.83 34.57
± 1.63 33.53
± 1.64
UrineNS
4.35 ± 0.69 4.76 ± 0.72 4.72
± 0.37 4.05
± 0.65
TotalNS
42.16 ± 3.54 41.35 ± 0.46 39.29
± 1.27 37.58
± 1.45
Calcium retention
Retention, g/dNS
19.95 ± 3.54 20.76
± 0.46 22.82
± 1.27 24.54
± 1.45
% intakeNS
32.12 ± 5.70 33.42 ± 0.74 82.91 ± 0.39 85.87 ± 1.95
% absorbedNS
79.97 ± 6.13 81.52 ± 2.28 82.91 ± 0.39 85.87
± 1.95
NS – Non Significant
79
calcium retention expressed as either g/d, as per cent of intake or as per cent absorbed were higher in T4 and lower in T1
when compared to other treatments. However, the differences between treatments were not significant (P>0.05).
4.4.4 Phosphorus balance
Phosphorus intake, outgo and retention in buffalo bulls fed total mixed ration supplemented with exogenous
fibrolytic enzyme and/or live yeast culture are presented in Table 14. The phosphorus excretion through faeces, urine or
total phosphorus excretion (g/d) was higher in T1 and lower in T4 when compared to other treatments. However, no
significant (P>0.05) differences were observed between the treatments. All the animals were in positive phosphorous
balance. The phosphorous retentions (g/d) were 15.28, 15.77, 16.02 and 16.48 in T1 T2, T3 and T4, respectively. The
phosphorus retention (g/d) was higher in T4 and lower in T1 when compared to other treatments but, the differences between
treatments were not significant (P>0.05). The phosphorous retention expressed as either g/d, as per cent of intake or as per
cent absorbed were higher in T4 and lower in T1 when compared to other treatments. However, no significant (P>0.05)
differences between treatments were observed.
80
Table 14: Phosphorus utilization in buffalo bulls fed total mixed ration supplemented with exogenous fibrolytic
enzymes and/or live yeast culture
T1 T2 T3 T4
Phosphorus intake, g/d 29.23 29.23 29.23 29.23
Phosphorus outgo, g/d
FaecesNS
10.99 ± 0.27 10.58
± 0.62 10.52
± 0.29 10.35
± 0.79
UrineNS
2.95 ± 0.25 2.88
± 0.31 2.69
± 0.24 2.40
± 0.27
TotalNS
13.94 ± 0.16 13.46
± 0.65 13.20
± 0.26 12.75
± 0.67
Phosphorus retention
Retention, g/dNS
15.28 ± 0.16 15.77
± 0.65 16.02
± 0.26 16.48
± 0.67
% intakeNS
52.29 ± 0.53 53.95
± 2.21 54.83
± 0.88 56.37
± 2.29
81
% absorbedNS
83.86 ± 1.19 84.53
± 1.58 85.66
± 1.16 87.31
± 1.19
NS – Non Significant
4.4.5 Plane of nutrition of buffalo bulls
The data on plane of nutrition of buffalo bulls fed total mixed ration supplemented with exogenous fibrolytic enzyme
and/or live yeast culture is presented in Table 15. The DM intake expressed as per cent of body weight was similar among
the treatments. The DCP content expressed as % in the diet consumed or as intake (kg/d) was marginally higher in T4 as
compared to other treatments. However, the differences between the treatments were not statistically significant (P>0.05).
Similarly, the TDN content expressed as % in the diet consumed or as intake (kg/d) was higher in T4 and lower in T1 when
compared to other treatments but the differences between treatments were not significant (P>0.05). Further, the estimated
DE and ME intakes (M cal/d) also followed the same trend. The DM, DCP and TDN intakes expressed as g/kg W0.75
were
similar among the treatments. Similarly, the ME intake (M cal/kg W0.75
) and the protein energy ratio (g/M cal) were
comparable among the treatments.
82
82
Table 15: Plane of nutrition of buffalo bulls fed total mixed ration supplemented with exogenous fibrolytic enzymes
and/or live yeast culture
NS – Non Significant
Treatmen
t
Avg.
B .Wt
W
kg0.75
DMI
(kg/d
)
DMI
as
%
B.Wt
.
DCP TDN DE
IntakeN
S
(Mcal)
ME
IntakeN
S
(Mcal)
Intake /unit W kg0.75 DCP
(g) /
MENS
(Mcal
)
% in diet
consumedN
S
IntakeN
S
(kg/d)
% in diet
consumedN
S
IntakeN
S
(kg/d)
DMNS
(g)
DCPN
S
(g)
TDNN
S
(g)
MENS
(Mcal
)
T1 376.6
3
85.4
0
6.03 1.62
7.96
0.48
56.67
3.42
15.06
12.36
71.0
6
5.67 40.32
0.15
38.84
T2 376.6
3
85.5
5
6.03 1.70
8.19
0.49
58.67
3.54
15.59
12.79
70.9
1
5.82
41.60
0.15
38.60
T3 376.6
3
85.6
2
6.03 1.67
8.24
0.50
58.89
3.55
15.65
12.84
70.8
6
5.84
41.69
0.15
38.69
T4 376.6
3
85.6
4
6.03 1.68
8.25
0.50
59.19
3.57
15.73
12.91
70.8
4
5.84
41.86
0.15
38.58
ICAR
1998
350.0
0
80.9
2 5.0 1.5 - 0.23 - 2.7 11.46 9.4
61.7
9 2.84 33.87 0.12 24.48
Kearl,
1982
350.0
0
80.9
2 5.7 1.6 - 0.23 - 2.6 11.59 9.5
70.4
4 2.85 32.13 0.12 23.75
83
i
CHAPTER – VI
SUMMARY
In the present study, a total mixed ration (TMR) containing 12 % CP was
prepared using groundnut haulms as roughage source and was supplemented with
exogenous fibrolytic enzymes (EFE) and/or live yeast culture to form the 4
dietary treatments. In 4 x 4 LSD, four graded Murrah buffalo bulls (avg. b. wt.
377.05 ± 43.36 kg) were randomly allotted to four dietary treatments viz. TMR
with R: C ratio of 70: 30 (T1), T1 supplemented with EFE @ 15 g/animal/day (T2),
T1 supplemented with yeast culture @ 10 g/animal/d (T3) and T1 supplemented
with EFE @ 15 g/animal/day and yeast culture @ 10 g/animal/d (T4) and
evaluated for their effect on in vitro digestibility, rumen fermentation pattern,
mineral balances and nutrient utilization in buffalo bulls.
Both groundnut haulms as well as groundnut haulms based total mixed
ration were evaluated for their chemical composition and cell-wall constituents. In
vitro digestibilities (%) of TMR supplemented with EFE and/or live yeast culture
were in the range of 50.97 to 57.02; 53.98 to 60.57; 49.15 to 57.28 and 46.42 to
55.00 for DM, CP, NDF and ADF, respectively. The in vitro digestibility (%) of
DM, CP, NDF and ADF were lower (P<0.01) in T1 when compared to T2, T3 or
T4. Further, the in vitro digestibility (%) of DM, CP, NDF and ADF increased
linearly from T2 to T4. However, no significant (P>0.05) differences were
observed between T2 and T3, T2 and T4 and T3 and T4.
Rumen fermentation studies conducted using graded Murrah buffalo bulls
revealed that rumen pH values were highest at 0 h (before feeding) and declined
ii
to minimum by 4 h post feeding while TVFA concentration (meq/L) was the
reverse of pH which increased gradually, peaked at 4 h post feeding and then
declined irrespective of the treatment. Further, NH3-N, total N, TCA-insoluble N,
residual N and food and protozoal N (mg/100ml SRL) also reached peak 4 h post
feeding and later followed a decreasing trend in all the treatments. The present
study indicated that supplementation of EFE in TMR (T2) had no effect (P>0.05)
on rumen pH and food and protozoal N concentration while it increased (P<0.01)
the TVFA, NH3-N, total N, TCA-insoluble N, and residual N when compared to
T1. Moreover, the present study also revealed that supplementation of yeast
culture in TMR increased (P<0.01) rumen pH and the concentrations of TVFA,
NH3-N, total N, TCA-insoluble N, and residual N while it had no effect (P>0.05)
on food and protozoal N in buffalo bulls. Furthermore, the present study indicated
that supplementation of EFE and/or live yeast culture in TMR (T4) increased
(P<0.01) rumen pH and the concentrations of TVFA, NH3-N, total N, TCA-
insoluble N, residual N and food and protozoal N in buffalo bulls as compared to
the control.
The digestibility coefficients of DM ranged from 55.21 to 58.16, OM from
59.88 to 62.62, CP from 60.31 to 62.51, EE from 66.79 to 69.61, CF from 48.52
to 51.86, NFE from 66.70 to 69.15, NDF from 50.60 to 55.19, ADF from 47.18 to
52.50, hemi-cellulose from 63.85 to 66.85 and cellulose from 59.16 to 63.78 in
buffalo bulls fed total mixed ration supplemented with EFE and/or live yeast
culture. The digestibility of DM, OM, CP, EE, NFE, NDF, ADF, hemi-cellulose
and cellulose increased linearly from T1 to T4 but the differences between
treatments were not significant (P>0.05). The present study revealed that
iii
supplementation of EFE and/or live yeast culture in TMR had no effect (P>0.05)
on the digestibility of gross nutrients and fibre fractions.
The balance studies indicated that all the buffalo bulls were in positive
balance for N, Ca and P. The N, Ca and P excretion through faeces, urine and total
outgo (g/d) were higher in T1 and lower in T4 but the differences between
treatments were not significant (P>0.05). Further, the present study revealed that
supplementation of EFE and/or live yeast culture in TMR had no effect (P>0.05)
on nitrogen, calcium and phosphorous retentions expressed either as g/d or as %
intake or as % absorbed in buffalo bulls.
The average DMI of buffalo bulls expressed as kg/d or as % BW was
comparable among the treatments. The present study indicated that
supplementation of EFE and/or live yeast culture in TMR had no effect (P>0.05)
on DCP and TDN content expressed as % in the diet consumed or as kg/d.
Furthermore, the DM, DCP, TDN and ME intakes per kg W0.75
were similar
among the treatments and were higher than the values recommended by ICAR
(1998) and Kearl (1982) standards. Similarly, the DCP: ME ratio was comparable
among the four dietary treatments.
Based on the results obtained from the present study the major findings are
as follows:
1. In vitro studies indicated that supplementation of EFE and/or live yeast
culture in TMRs improved the in vitro digestibility (%) of nutrients.
iv
2. Rumen fermentation studies indicated that supplementation of EFE and/or
live yeast culture in TMR improved the concentrations of total VFA, NH3-N
and N fractions in the rumen.
3. The DMI in buffalo bulls was higher than that recommended by ICAR
(1998) and Kearl (1982) indicating that the diets are palatable and that
supplementation of exogenous fibrolytic enzymes and/or live yeast culture
had not affected the palatability.
4. Supplementation of EFE and/or live yeast culture in TMR had no effect
(P>0.05) on the digestibility of gross nutrients and fibre fractions.
5. All the animals were in positive balance for N, Ca and P. Further,
supplementation of EFE and/or live yeast culture in TMR had no effect
(P>0.05) on retentions of N, Ca and P in buffalo bulls.
6. Plane of nutrition of buffalo bulls during the study period revealed that
supplementation of EFE and/or live yeast culture has no effect on DCP and
TDN (%) and on DM, DCP, TDN and ME intakes per kg W0.75
.
Thus, it is concluded that in vitro digestibility of nutrients increased with
supplementation of EFE and/or live yeast culture to groundnut haulms based
TMR while in vivo studies conducted in buffalo bulls revealed no effect on DM
intake and on the digestibility of gross nutrients and fibre fractions. However,
rumen fermentation studies conducted in buffalo bulls revealed improvement in
the concentrations of TVFA, NH3-N and N fractions.
v
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