evaluation of total mixed rations supplemented with exogenous fibrolytic enzymes and / or live

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


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


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


total mixed rations

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


Groundnut haulms 81


total mixed rations 82


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



66

8 TCA insoluble protein nitrogen (mg/100ml SRL) in



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



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


yeast culture

76

Table

No. Title Page No.

14 Phosphorus utilization in buffalo bulls fed total mixed


yeast culture

78

15 Plane of nutrition of buffalo bulls fed total mixed


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

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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).

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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


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

110

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


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).

114

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.


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.

116


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

117

the apparent digestibility of nutrients. Further, supplementation of either SC or

MOS alone or in combination had no affect on DMI.

118

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.

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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



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



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


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


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


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


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


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


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


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


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


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


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


151

Table 13: Calcium utilization in buffalo bulls fed total mixed ration supplemented with exogenous fibrolytic enzymes


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


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


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


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



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


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


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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Documents

evaluation of total mixed rations supplemented with exogenous fibrolytic enzymes and / or live