STUDY OF FACTORS AFFECTING QUALITY OF SILAGE AS A

COMPONENT OF TOTAL MIXED RATION ON GROWTH AND

PRODUCTION PERFORMANCE IN NILI-RAVI BUFFALOES

BY

RAFI-UDDIN

2004-VA-152

A THESIS SUBMITTED IN THE PARTIAL FULFILLMENT OF THE

REQUIREMENT FOR THE DEGREE

OF

DOCTOR OF PHILOSOPHY

IN

LIVESTOCK MANAGEMENT

UNIVERSITY OF VETERINARY & ANIMAL SCIENCES,

LAHORE

2016

To,

The Controller of Examinations,

University of Veterinary and Animal Sciences,

Lahore.

We, the Supervisory Committee, certify that the contents and form of the thesis, submitted

by Mr. Rafi-Uddin, have been found satisfactory and recommend that it be processed for further

evaluation by the External Examiner (s) for the award of the Degree.

CHAIRMAN __________________________________________

(PROF. DR. MUHAMMAD ABDULLAH)

MEMBER ___________________________________________

(PROF. DR. KHALID JAVED)

MEMBER ___________________________________________

(PROF. DR. MAKHDOOM ABDUL JABBAR)

i

DEDICATION

I DEDICATE THE FRUIT OF THIS HUMBLE EFFORT

TO

MY BELOVED PARENTS (LATE)

ii

ACKNOWLEDGEMENTS

All prays to almighty “Allah” Who provided means and opportunities on my way and gave

me will, strength and health to accomplish this task, who gave me ambassadors at earth for

intellectual interaction with them.

In the first place, I would like to gratefully acknowledge Prof. Dr. Muhammad Abdullah

for his enthusiastic supervision, advice, and guidance from the very early stage of my doctoral

program as well as giving me extraordinary experiences throughout the work. Above all and the

most needed, he provided me unflinching encouragement and support in various ways.

It is an honor for me to express gratitude to other committee members; Prof. Dr. Khalid

Javed, and Prof. Dr. Makhdoom Abdul Jabbar for their patient guidance, encouragement and

excellent advice throughout the study period. I am also grateful to other teachers in the faculty of

Animal production and Technology; Dr. Jalees Ahmad Bhatti and Dr. Nisar Ahmad, whose helpful

suggestions reduced ambiguity in my work under friendly environment.

I would like to express my gratitude to the Vice Chancellor, University of Veterinary and

Animal Sciences Lahore for giving good environment for scholars to flourish. I am also grateful

to Secretary, Livestock and Dairy Development (L&DD) Baluchistan for granting me study leave.

Their cooperation enabled me to go across the course of higher education without financial crisis.

It is difficult to overstate my gratitude to all the scientists of the world and to those who

helped build the equipments that allowed me to run my experiments; without them, I could not be

succeeded to write this dissertation. I would be glade to extend my thanks to Dr. Nawaz Saeed

(Ex. Chief Research Officer BRI, Pattoki), Dr. Ihsanullah (Farm Superintendent) and Mr.

Muhammad Asim (Farm Manager), They extended their cooperation to me for supply of research

material and facilities needed at my research site, Buffalo Research Institute Pattoki, District

Kasur.

During this work I have collaborated with many colleagues and workers for whom I have

great regard and I wish to extend my warmest thanks to all those who have helped me with my

work. I am thankful to research fellows: Mr. Mehtab Ahmad, Mr. Rahman Ullah Mohmand, Mr.

Muhammad Ramadan and Mr. Mir Ahmad Khan. I can’t forget dedication and hard work of Mr.

iii

Muhammad Imran, Abdul Shaboor, Noor Ahmad and Muhammad Yaseen. They were physically

involved while handling animals.

I would like to express my profound gratitude to my beloved Parents (Late), elder brothers,

Prof. Allauddin, Mr. Sallahuddin and younger brother Mr. Sharafuddin. I would also like to

mention Abgina Khan, Khushal Khan, Muhammad Abdullah and little Ayika Khan, my children,

who teased me a lot during my studies, lots of love and prayers for them. I feel guilty if I don’t

pay my thanks to my beloved wife who acknowledged, exhibited great patience and sacrificed her

settled life for my doctoral program.

Finally I would like to appreciate Higher Education Commission, Pakistan for providing

Indigenous Scholarship which helped me a lot in completion of my studies and providing an

opportunity for conducting six months research in University of Sydney, Australia through IRSIP.

May Almighty Allah shower his blessings on all of us…!!

Rafiuddin kakar

iv

TABLE OF CONTENTS

Sr.NO. Title Page No.

Dedication i

Acknowledgement ii

Table of contents iv

List of Tables vi

Abbreviations vii

Chapters

1 Introduction 1

2 Review of Literature 4

2.1. Stage of maturity on quality of cereal silages 4

2.2 Silo types effect on quality characteristics of cereal silages 9

2.3 Additive effects on fermentation dynamics of cereal silages 11

2.4 Feeding effects of cereal silages on production performance of dairy

animals

17

2.5 Literature cited 27

3 Experiment 1 33

3.1 Abstract 33

3.2 Introduction 34

3.3 Materials and methods 35

3.4 Results 38

3.5 Discussion 39

3.6 Literature cited 42

4 Experiment 2 48

4.1 Abstract 48

4.2 Introduction 49

4.3 Materials and methods 50

4.4 Results 53

4.5 Discussion 54

4.6 Literature cited 56

5 Experiment 3 61

5.1 Abstract 61

5.2 Introduction 62

v

5.3 Materials and methods 63

5.4 Results 67

5.5 Discussion 68

5.6 Literature cited 70

6 Experiment 4 76

6.1 Abstract 76

6.2 Introduction 77

6.3 Materials and methods 78

6.4 Results 79

6.5 Discussion 80

6.6 Literature cited 82

7 Experiment 5 86

7.1 Abstract 86

7.2 Introduction 87

7.3 Materials and methods 87

7.4 Results 89

7.5 Discussion 90

7.6 Literature cited 93

8 Summary 99

vi

LIST OF TABLES

Sr.NO. Title Page No.

3.1 Date of sowing and harvest stage of maturity for three cereal fodders 45

3.2 Effect of maturity stages on chemical composition and nutritive value

of silages

45

3.3 Fermentation characteristics and IVDMD of silages at different stage

of maturity

46

3.4 Effect of maturity stages on physical quality of silages 47

4.1 Date of sowing and harvest for three cereal fodders 58

4.2 Effects of silo types on physical characteristics of cereal silages 58

4.3 Effects of silo types on fermentation characteristics and IVDMD of

silages

59

4.4 Effects of silo type on chemical composition of cereals silages 60

5.1 Date of sowing and harvest for three cereals fodders 73

5.2 Effects of additive on fliege score, IVDMD and LA of silages 73

5.3 Effects of additive on pH during fermentation kinetics of silage 74

5.4 Effects of inclusion level of additive on chemical composition of

silages

75

6.1 Nutritional composition of different roughages fed to buffalo calves 84

6.2 Nutrients intake and weight gain of buffalo calves fed different roughages

84

6.3 Nutrients digestibility of different roughages fed to buffalo calves 85

7.1 Ingredients and chemical composition of four experimental TMR (DM

basis)

96

7.2 Nutrient intake (kg) and digestibility of maize fodder and its silage

based TMR

97

7.3 Milk yield and milk composition of four experimental groups 98

vii

ABBREVIATIONS

ADF Acid Detergent Fiber

BW Body Weight

CP Crude Protein

DM Dry Matter

DMD Dry Matter Digestibility

DMI Dry Matter Intake

GE Gross Energy

IVDMD In-vitro Dry Matter Digestibility

Kg Kilogram

Mcal Mega Calories

ME Metabolizable Energy

N Nitrogen

NDF Neutral Detergent Fiber

NRC National Research Council

OM Organic Matter

PARC Pakistan Agricultural Research Council

SE Standard Error

TDN Total Digestible Nutrient

TMR Total Mixed Ration

WSC Water Soluble Carbohydras

1

CHAPTER 1

INTRODUCTION

Buffalo (Bubalus bubalis) is a fundamental provider of milk, meat, draught power, fuel

and leather production in some developing countries of Asia. Buffaloes could be categorized into

Asian and Mediterranean buffaloes. Asian buffalo are further divided into binary category known

as the water and Marsh (Swamp) buffalo types. The water buffalo (Riverine) having 50

chromosomes while Swamp buffalo have 48, with unique physical structure and behavior meaning

they are reared for different purposes. Riverine buffaloes are mainly found in India, Pakistan and

in some countries of western Asia, primarily used for milk and meat production (Sarwar et al.,

2002a, b). Swamp buffaloes are mainly found in South East Asian countries, primarily used for

draught power but are also used to produce meat and small quantity of milk (Sarwar et al., 2009).

Water buffalo is the major dairy animal of Pakistan contributing about 67% of overall milk

produced in the country (Afzal et al., 2007). It can produce about 8-12 liter/day for 280 day

lactation length with 6 to 9% milk fat (Sarwar et al., 2002. Afzal et al., 2007). Because of its higher

milk fat contents, that milk of buffalo is preferred than camel and cow milk and get higher price

in milk markets of Asian countries (Sarwar et al., 2002 b; Khan et al., 2008a). Current population

of buffaloes in Pakistan is approximately 27.3 million heads and the country getting second

position in the world after India (Khan et al., 2007). Nili-Ravi buffalo is the well-known

widespread dairy breed of the country, which constitutes about 76.6% of the total buffalo

population in Pakistan mainly reared in the Punjab province (Batth et al, 2012). However, milk

yield is still much less compared to developed dairy breeds like Holstein and Jersey, largely due

to random breeding, inefficient reproductive management, poor nutrition and fluctuation in quality

feed supply (Sarwar et al., 2009).

Field grown cereals especially maize (Zee maize), sorghum (Sorghum bicolor), millet

(Pennisetum americanum), oat (Avena sativa) and legumes including lucerne (Medicago sativa)

Introduction

2

and berseem (Trifolium alexanderinum) forages contribute a key role in the agricultural economy

of developing countries by providing economical feedstuff. Forage feeding alone contribute

approximately 70% of the total cost of livestock production in the developing countries of the

world specially Asia including Pakistan (Bhatti et al., 2009). Grown area under fodder production,

in the country are near to 10.5 % of the total cultivated area of 22.5 million hectares. The share in

cultivated area of Punjab, Sindh, KPK and Baluchistan are as 82.57, 11.51, 4.49 and 1.47%

respectively (Fodder research program, PARC. 2016). In the country cultivated area under

numerous fodder crops is roughly 2.35 million hectares with 52.92 of fodder production per year.

Per hectare fodder production is 22.51 million tons approximately (Agric. Statistic. of Pakistan

2009-10). Fodder supply for livestock particularly large ruminants feeding is about 54-60% lesser

then required demand, probably due to low per acre yield and preference of cash crop cultivation,

this constraints minimized the area for fodder production (Sarwar et al., 2002b). This shortage of

supply is joined with the reduction in area under fodder crops by 2% per decade after, mostly due

to urbanization (Sarwar et al., 2002b).

The performance of dairy animals depends largely on the continued availability of forage

quality in adequate amounts. Subsequently, the fundamental limitation on profitable livestock

production in under-developed and developing countries is the lack of valuable quality forage

(Sarwar et al., 2002). In many under progress countries due to increasing human necessity for food,

only limited cultivated land can be allocated to fodder production. Moreover, in this region, the

low performance for period’s acres of forage and fodder shortages, one is during the summer

months and in second place in the winter months, further exacerbate the situation. (Sarwar et al.,

2002). While, in reset of year forage is abundant and leftovers together in the fields. The use of

this additional forage can bridge the gap between supply and demand during periods of dearth.

Through silage technology the conservation of extra cereals and grasses forage in the abundant

season could help to reduce irregular troubling pattern throughout the year. Cereals and grasses

Introduction

3

forages are being used widely worldwide for making silage due to its relatively low buffering

capability and higher concentration of fermentable water soluble carbohydrates ((Bolsen et al.,

1996)). Therefore, the decline in pH is rapid, and the final pH is mostly low as compared to lucerne

and berseem fodders (Bolsen et al., 1996). At all cereals and grasses fodders that have sufficient

fermentable carbohydrates (WSC) can be silage, but the most popular crop for silage making is

corn (Woolford, 1984). Nonetheless, oat, sorghum, barley, millet, and grasses mott and Jambo

were used for silage making throughout the world (Tauqir et al., 2007). Deterioration in the quality

of forage during ensiling period of silage occurs during aerobic phase. Though, the preservation

of excess fodder and making the free land for planting are advantages subsequent silage ensilation.

However, due to lack of research and scientific literature on silage making and its benefits

for livestock production, limits the propagation of silage. Furthermore, continuous feeding fresh-

green fodder (legumes and grasses) through “cut and carry system” is still contributing

significantly to the nutrition of animals. Due to these constraints in developing countries of South

Asia, dairy producers trust in a strong myth that silage feeding could hinder productivity of

animals, in-terms of growth rate and milk yield (Khan et al., 2006c).

In Pakistan silage production is in its infancy stage and lake of awareness among farmers

regarding silage feeding and its preservation is limited. Moreover, the aim of this study was

developed ideas and acceptability of feed silage and among small-scale milk producers and fodder

collection to its good state of maturity, ideal silage technology (silo) ensilation and advantages of

silage feed for abundance, as well as the period of scarcity of dairy practices circa on growth and

milk production Nali-Ravi buffalo.

4

CHAPTER 2

REVIEW OF LITERATURE

The silage production is relatively new practice in Pakistan. Various aspects of crop

production as well as storage conditions affect the quality of silage. In the following paragraphs

we have reviewed the effect of stage of plant maturity, types of silos and levels of bacterial

inoculation on fermentation characteristics, composition, and physical quality of silages. The

previous studies conducted to evaluate the productive performance of buffaloes fed silage verses

fresh fodder has also been presented in this section.

2.1. Stage of maturity on quality of cereal silages

Stage of plant maturity is one of the most important factor influencing nutritional quality

and fermentation characteristics of cereal silages (Khan et al., 2011). However, there is conflicting

evidence on the ideal stage of harvesting for silage making of cereal forages. Supreme stage of

forage to maintain nutritive value and quality of silage varied in literature of the previous

researchers.

St. Pierre et al. (1987) Investigated the feeding value of maize silage harvested at the milk

or dough stage, or after one, two, or five frosts (number of days between harvest or interval, after

dough stage) in lactating Holstein cows. The neutral detergent fiber (NDF) and dry matter (DM)

contents of silage significantly increased from 59.0 to 65.9 and 23.2 to 45.0 respectively as the

maturity advanced form milk stage to after five frost. Dry matter intake increased (14.5 to 16.5

kg/day) and 4% fat-corrected milk increased (19.6 to 19.7 kg/day) as maturity of the silage

increased up to the silage harvested after two frosts and then declined for the silage harvested after

five frosts. Gross energy apparent digestibility decreased from 64.9% for milk stage silage to

60.6% for silage from corn harvested after five frosts. Concluded, that maize crop should be

Review of Literature

5

harvested for quality silage at its optimum stage of maturity (34% DM) after second frost and

obtained maximum production for a profitable dairying.

Khan et al. (2011) carried out a laboratory silos based experiment on three stages of

physiological maturity i.e., pre heading, heading and milk stage of maize (Zea mays), sorghum

(Sorghum bicolor) and millet (Pennisetum americanum) silages. The results showed that the dry

matter (DM), neutral detergent fiber (NDF), acid detergent fiber (ADF) and lactic acid contents

increased (P<0.05) in all three fodder silages with advancing age. However, crude protein (13.30

to 8.82., 8.90 to 6.06 and 8.84 to 5.18), total digestible nutrients (63.83 to 58.12, 61.97 to 56.93,

and 60.59 to 55.75) and metabolizable energy contents (2.39 to 2.15, 2.31 to 2.08 and 2.24 to 2.02)

decreased (P<0.05) respectively, in all three silages with the advancement of their growth. The

water soluble carbohydrates and pH values of silages prepared from three fodders decreased

(P<0.05) as the plants advanced in age. And recommended that “milk stage” of physiological

maturity was the best stage for silage making of cereals fodder.

Russell. (1986) investigated the weekly growth intervals of harvest date, from 3 weeks pre

to 5 weeks post physiological maturity on the ensiling characteristics of maize stovers silage. The

results showed a linearly (P<0.01) decreased in crude protein (CP) contents (87.6 to 77.9 g/kg

DM) and quadratic increased in DM (295, 282 and 311 g/kg) from early to later harvested stovers.

Though, the changes in the concentrations of neutral detergent fiber (NDF; 626.0 679.0 g/kg) and

acid detergent fiber (ADF; 354.4 405.1 g/kg) were inversely related to those of CP. While, lactic

acid in the stovers silages increased (29.8 to 30.0 g/kg) with later harvest. Therefore, heifers fed

on the early-harvested silage tended to have greater daily gains (0.47kg/day) as compare to later

harvested (0.43kg/day) and required less feed per kg gain (P~0.05) than did heifers fed on the late-

harvested stovers silage.

Review of Literature

6

Firdous et al. (1996) also summarized the effects various growth stages at weekly intervals

on in-vitro digestibility of whole plant and its morphological fractions (leaf and stem) of maize

cultivar. The results showed that in-vitro dry matter digestibility (IVDMD) declined significantly

(P<0.01) in whole maize plant (75.82, 70.30, 67.35 and 65.69), as well as in leaf (77.04, 75.75,

73.98 and 69.94) and stem (66.82, 62.02, 59.45 and 58.52) fractions respectively with advancing

stage of maturity corresponding to early growth, flowering, milk/dough and mature stage.

Similarly, IVDMD of neutral detergent fiber (NDF) and acid detergent fiber (ADF) were also

significantly decreased in whole plant and its subsequent leaf and stem fractions with advancing

growth. Results revealed that maize fodder should be harvested at its flowering stage of maturity

(between 8th - 9th week of age) to obtain better digestibility and harvest better quality nutrients for

livestock feeding.

Edmisten et al. (1998) evaluated the effects of six stages of maturity (vegetative, boot,

heading, milk, soft dough and hard dough) in small grain cereals (barley, oat, rye, and wheat) on

nutritive quality as a replacement for corn silage. The IVDMD all species generally decreased

from the vegetative (765-854 g/kg) through the milk stage (505-662 g/kg) and then remained

unchanged or increased slightly through hard dough. While, IVDMD of ensiled barley was (805,

779, 736, 634, 633 and 584 g/kg) respectively for vegetative, boot, heading, milk, soft dough and

hard dough stages of maturity a similar pattern were also observed for oat, rye, and wheat ensiled

forages. The NDF, and ADF fractions usually increased from the vegetative to milk stages and

remained unchanged or increased slightly through the hard dough stage. The ADF and lignin are

negatively associated with forage digestibility while NDF values are negatively related to dry

matter intake. Crude protein content generally decreased from the vegetative through milk stages

and then leveled off or decreased slightly through the hard dough stage. Results of study suggested

that the vegetative or boot stages would provide high quality forage for grazing or green crop

operations.

Review of Literature

7

Khan et al. (2012) ensiled maize forage at 300, 340, 380, and 420 g/kg DM contents of

fresh weight denoted by (MS30, MS34, MS38 and MS42) and fed to dairy cows in combination

with concentrate. Sixty-four multiparous Holstein-Friesian dairy cows in their first week of

lactation were assigned to the four dietary treatments according to a randomized complete block

design (RCBD). Corn silages were offered ad libitum as part of a basal forage, whereas the

concentrates were given at the rate of 8.5 kg of DM/Cow per day during the experimental period.

Results shown that dry matter (DM), crude protein (CP) and energy intakes did not differ across

the maize silages harvested at different DM contents MS30 to MS42. While, the intake of NDF

and ADF were decreased 366, 350, 345 and 341 g/kg; and 212, 202, 198 and 196 g/kg respectively,

with increasing maturation. Milk yield and composition was not different across the corn silages,

except for fat content lowered in MS42 (1.60 kg/d) compared to MS34 (1.70 kg/d) and MS38 (1.70

kg/day).

Rinne. (1997) studied 1st cut mixed herbage (timothy “Phleum pretense” and meadow

fescue “Festuca pratemis” silages harvested at four stages of maturity, 1) pre bloom, 2) early

bloom, 3) full bloom ,and 4) late bloom stage (approximately one-week intervals) from the same

sward. However, progressive maturity of herbage increased neutral detergent fiber (NDF; from

409, 497, 579 and 623 g/kg of DM). While, decreased CP content (29.9, 26.7, 18.7 and 17.4 g/kg)

associated with decreased digestibility of organic matter OM (0.821, 0.816, 0.758 and 0.747 g/kg)

respectively of silages in order of increasing maturity in the field. N intake decreased significantly

(P< 0.01; 167.5 to 118.0 g per day) with advancing maturity of grass. Early harvest of herbage

ensured high production level by high OM digestibility of silage. While, very early harvest cannot

be recommended due high nitrogen losses from the animals.

Hussain et al. (2002) studied the effects of harvesting intervals (maturity) of oat forage

harvested on days 70, 85, 100 and 115 of planting to investigate nutritionalionl quality of fodder.

Review of Literature

8

The results revealed that CP contents decreased significantly from 13.06 to 8.02 per cent While,

in contrast crude fiber increased 23.04 to 24.63 per cent with advancing age of oat. Concluded,

that minimum green forage and dry matter yield with superior quality forage as determined by

higher crude protein and lower crude fiber contents were observed when the crop was harvested

or grazed after 85 days of sowing.

Khorasani et al. (1997) studied maturity stages from boot to soft dough on chemical

composition of barley, oats, and triticale cereal crop silages. Results revealed that the relationship

between DM content and stage of maturity of crops were significant DM content of crops increased

as maturity advanced. Overall DM percent increased from an average of 13% for first harvest (at

boot stage) to an average of 41.9% (± 4.9 SD) for the last harvest (at soft-dough stage). While, a

linear and quadratic (P < 0.05) correlations were observed between the CP content of cereal grain

crops and advancing maturity. CP concentration declined from an average of 23% at the boot stage

to 15% at the soft-dough stage. The relationship between maturity and NDF concentration was

linear for alfalfa, whereas this relationship for the cereals was curvilinear. Results indicated that

by harvesting small grain cereals at the proper stage of maturity, the fiber and CP concentrations

in cereal forages can be manipulated to meet the feeding quality standards and getting maximum

levels of growth and milk production.

Mustafa and Seguin. (2003) carried out an experiment to investigate the effects of oat silage

(ensiled in mini-silos) harvested at boot or soft dough stage of maturity on digestibility and

chemical composition. Two ruminal cannulated lactating Holstein cows were used to determine

in-situ ruminal nutrient degradability. Results of the in-situ incubation experiment indicated that

oat harvested at the soft dough stage had lower ruminal dry matter (60.6 vs. 66.4%). CP (81.3 vs.

88.7%) and NDF (35.4 vs. 42.2%) degradability than oat harvested at the boot stage. However,

chemical characteristics of oat silage after 45 days of ensiling at the boot stage contained more

crude protein (CP) and less starch than that harvested at the soft dough stage. While, stage of

Review of Literature

9

maturity had no effect on neutral detergent fiber (NDF) and acid detergent fiber (ADF) of oat

silage. It was concluded that chemical composition and ruminal nutrient degradability of oat silage

are significantly influenced by stage of maturity.

Giardini et al. (1976) carried out an experiment in a factorial design, among three stages of

maturity of maize silage (milk stage, dough stage and late dough stage with 24-28-39% DM at

harvest) and its feeding value in young Polish Holstein bulls. The best results were attained forages

were harvested at 39% DM content in an all-maize silage ration. While earlier harvests stage

having 24% and 28% DM gave rations with a higher feed efficiency of the DM but were not

feasible due low dry matter yield with increasing costs of feeding per hectare.

2.2. Silo types Effect on quality characteristics of cereal silages

In recent years or before in the world most of the silage quality experiments have been

carried out in laboratory scale silos (LSS) under confined environmental condition. Whereas, in

contrast big scale silo (BSS) research is needed. Which ultimately expose to varying environmental

factors including seasonal temperature, finally affect silage quality in terms of pH, and chemical

composition (Kızılsimsek et al., 2005).

Therefore reviews of researcher about silo types are presented as, Kızılsimsek et al. (2005)

who studied both winter (expt: 1) and summer (Expt: 2) growing fodder crops. Ensiled in

laboratory scale silo (LSS) and big scale silo (BSS) for 45 days of incubation period, with loading

bulks of 10 kg and 3 tons of fresh material, respectively. The results showed that’s silo type had a

significant effect on silage structure score (3.33 and 3.71, respectively for BSS and LSS) in the

first experiment whereas it affected the color score (3.47 and 2.97, for BSS and LSS) of silage in

the second experiment. Moreover, significantly higher mean smell scores (4.83 and 4.43 of expt:

1 and 6.10 and 4.73 for expt: 2) were also observed in BSS followed by LSS. While, in terms of

fermentation characteristics significantly the lowest pH value (3.80 and 3.95 expt: 1 and 4.16 and

Review of Literature

10

4.32 expt: 2, respectively) were also observed in BSS followed by LSS silo. Concluded that silage

in BSS were better fermented than LSS.

Mtengeti et al. (2014) also carried out an experiment in a factorial design of 2 x 2 x 2 with

two replications to investigate the effects of processing, additive and silo type on the quality of

elephant grass silage. Four treatments were imposed on the forage before it was ensiled in either

earth pits or concrete silos. After 90 days of incubation period the DM losses did not differ

significantly between the silo types. However, the CP content was significantly (P < 0.0001; 61.6

vs 54.4 g/kg DM) higher in earth pit than concrete silos. Significantly (P < 0.05) lower pH and

higher in-vitro dry matter digestibility IVDM (4.28 vs 4.35 and 48.7 vs 43.5 g/kg DM,

respectively) were observed in earth pit as compared to concrete silo. The sensoric scores,

appearance, smell and texture of silage were significantly (3.1 vs 2.7, 2.7 vs 2.4 and 2.4 vs 1.9,

respectively) better for earth pit as compared to concrete silo. There was a non-significant effect

among the treatments when the animals were fed either earth pit or concrete silo silages. It can be

concluded that ensiling of elephant grass in earth pit silo can produce good quality silage.

A review paper submitted by Johnson et al. (2001) who investigated that type of silos are as trench,

bunker, bag, and upright has a significant impact on quality of silage in terms of DM and CP

contents during storage. He reported that DM loss was higher in bunker and trench silo compared

to bag and upright silos due to exposer of greater surface area to oxygen and environmental

temperature, thereby affect microbial fermentation inside the silo. Also find out inadequate

research suggests corn silage harvested at 38% DM was preserved well in a bag silo compared to

a bunker or trench silo. For obtaining better quality silage rapid filling of silo (bunker, trench) by

means of the progressive wedge technique has been progress fermentation and nutritive value of

resulting silage.

Review of Literature

11

Meeske et al. (2002) studied the effects of inoculants (Lactobacillus plantarum and

Pediococcus acidilactici as well as amylase) on maize silage ensiled in laboratory and bunker silos.

The inoculant did not result in a more rapid lowering of the pH or a more rapid lactic acid

production compared to untreated maize silage made in laboratory silos. Both the control and

inoculated maize silages were well preserved with a pH of 3.57 and 3.62 and lactic acid

concentration of 66 and 63 g/kg DM, respectively. The maize silages made in the bunker silos

were well preserved with a DM of 283 and 307 g/kg silage, pH of 3.50 and 3.51, lactic acid of 37.0

and 35.3 g/kg DM for the control and inoculated maize silage, respectively. The intake of untreated

and inoculated maize silage by Jersey cows was 7.6 and 8.4 kg DM/day for the control and

inoculant treatment, respectively. Milk production, milk composition, live weight and condition

score of Jersey cows was not significantly affected by the addition of the inoculant to maize silage.

2.3. Additive effects on chemical and fermentation dynamics of cereal silages

Homofermentative lactic acid bacteria present on maize plants prior to ensiling may be too

low to ensure rapid efficient preservation. In the USA 69% surveyed samples of maize crops

showed below 1000 colony forming units of lactobacilli per gram of fresh material (Speckman et

al., 1981). To obtain a high quality well fermented palatable silage, a rapid drop in pH is needed

to inhibit the growth of enterobacteria and clostridia (McDonald et al., 1981). This happens when

lactic acid bacteria (LAB) utilize water-soluble carbohydrates and produce lactic acid.

Worldwide used pre ensiling silage additives include biological inoculants, enzymes,

grains, molasses, urea, and acids (Weiss and Underwood, 2006). Minimum nutrients loss

especially protein and energy during fermentation period is the major goal of silage making, in

direction to attain this objective growth of acids producing bacteria should be stimulated during

initial anaerobic phase. Bacterial inoculant and enzymes specifically, commercial biological silage

inoculant was widely used to establish a desirable microbial flora in the ensiled forage.

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Nkosi et al. (2012) ensiled whole-crop sweet sorghum (Sorghum bicolor L. Moench) at

dough stage (323 g/kg DM) in 1 L anaerobic laboratory jars (silo) for 25 days of fermentation. To

investigate the effects of bacterial inoculation and cellulase on the fermentation quality of ensiled

forage. The treatments were i) no additive (control); ii) Lactobacillus buchneri (LB); iii)

Lactobacillus plantarum (LP); and iv) LB+E, a combination of LB and enzyme. Concluded that

inoculation significantly reduced pH in treatment iv) LB+E followed by iii) LP, LB ii) and control

3.57, 3.59, 3.62 and 3.90 respectively. And increased lactic acid content in (LP) followed by (LB)

and (LB+E) and lower in control (127, 95.3, 94.7 and 81.2 g/kg DM, respectively). However,

Inoculation also increased crude protein (39, 41.8, 41.4 and 40 g/kg DM) and decreased NDF (489,

438, 471 and 441 g/kg DM) and ADF (331, 297, 314 and 295 g/kg DM) in treatments i, ii, iii and

iv, respectively.

However, other researchers were also studied the use of additive to enhance silage quality

in response to bacterial inoculants application as, Jalc et al. (2009) who, studied the effect of three

bacterial probiotic inoculants (Lactobacillus plantarum, L. fermentum, and Enterococcus faecium

(coted as CCM 4000, LF2 and CCM 4231 respectively) on the nutritional characteristics and

fermentation quality of corn silage, under laboratory conditions. The inoculants were applied at a

concentration of 1.0×109 cfu/ml and treatment made as control (un-inoculated). The chopped corn

was ensiled in 40 plastic jars (1 L) divided into four groups (4×10 per treatment). All corn silages

had a low pH (below 3.55) and 83-85% of total silage acids comprised lactic acid after 105 days

of ensiling. The probiotic inoculants in the corn silages affected corn silage characteristics in terms

of significantly (P<0.05-0.001) higher pH (all treated silages had higher pH (3.48-3.54) than

untreated silage (3.44), numerically lower crude protein content compared to control silage.

However, the inoculants did not affect the concentration of total silage acids (acetic, propionic,

lactic acids) as well as in-vitro dry matter digestibility (IVDMD) of corn silages.

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Meeske et al., (2002) carried out an experiment to investigate the effect of lactic acid

bacterial inoculant, Maize-all (Lactobacillus plantarum and Pediococcus acidilactici as well as

amylase) on fermentation dynamics of the silage as well as the intake, milk production and milk

composition of Jersey cows fed maize silage diets. Maize fodder (PAN 6364) was harvested at the

half to three quarter milk line (DM 30%) and ensiled in laboratory and bunker silos. The inoculant

did not result in a more rapid lowering of the pH or a more rapid lactic acid production compared

to untreated maize silage made in laboratory silos. Both the control and inoculated maize silages

were well preserved with a pH of 3.57 and 3.62, a lactic acid concentration of 66 and 63 g/kg DM

respectively. The maize silages made in the bunker silos were well preserved with a DM of 283

and 307 g/kg silage, CP of 78.4 and 83.0 g/kg DM, pH of 3.50 and 3.51, lactic acid of 37.0 and

35.3 g/kg DM for the control and inoculated maize silage, respectively. The intake of untreated

and inoculated maize silage by Jersey cows was 7.6 and 8.4 kg DM/day for the control and

inoculant treatment, respectively. Milk production, milk composition, live weight and condition

score of Jersey cows was not significantly affected by the addition of the inoculant to maize silage.

Bilal. (2009) was conducted a laboratory silo based experiment to determine the best level

of additives (molasses and corn) applied at the rate of 0, 1, 3 and 5% of the forage dry matter

(DM). Mott grass was harvested at 45 and 60 days of its re-growth, chopped with an appropriate

particle length of ½ inches and filled in plastic boxes by mixing additives, with three replicates

each and kept at room temperature. Three silos of each treatment were opened at each fermentation

period (30, 35 and 40 days) for determination of pH and lactic acid contents. The results indicated

that mott grass cut at 45 days of its regrowth was the best to harvest maximum nutrients. The pH

decreased (from 4.83 to 4.07) and lactic acid increased (from 3.64 to 3.97) with level of additives

and fermentation periods. Dry matter and crude protein contents increased to some extent.

However, silage without additives showed the highest pH and low lactic acid, indicating the poor

quality silage. Similarly, a loss in DM and crude protein was observed in mott grass ensiled without

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additives. It was concluded that the use of additives such as molasses at the rate of 3% fodder DM

was found to be the best at 35 days fermentation period, to make quality mott grass silage.

Adesoji et al. (2010) carried out a study to investigate the impact of starter culture of

“Lactic acid producing bacteria (LAB) on quality of silage made from guinea grass (Panicum

maximum) silage. The chopped grass was inoculated with 10 ml of approximately 106 cfu/ml of

the L. plantarum, then ensiled into polyethylene bags. The results showed a rapid drop in the pH

of the inoculated silage (decreasing from 4.50 to 3.83) compared to the uninoculated silage (that

changed from 4.60 to 4.73) on day 15th and 30th respectively. The Crude protein (CP) contents

after 60 days of fermentation were also significantly higher in inoculated (9.84 g/100g DM) silage

as compare to control (7.86 g/100g DM). The ADF and NDF were also reduced in the inoculated

silage with 64.00 g/100 DM of NDF for the control silage at day 30 and 60.00 g/100 DM at the

same day for the inoculated silage. It is, therefore, concluded that inoculation of silage with homo-

lactic LAB increases fermentation as well as chemical composition of the ensiled forage.

Ozduven et al. (2010) revealed, the effects of lactic acid bacteria inoculant Sil-All (Alltech,

UK), enzymes (Global Nutritech, TR) and mixture of both on the fermentation, cell wall content,

and in vitro dry and organic matter digestibility characteristics of triticale (xTriticosecale

Wittmack) silages. Triticale (Pioneer -1188: Iowa, USA) was harvested at the milk stage of

maturity and inoculants were applied at rate of 6.00 log10 cfu/g of chopped fresh forage, then

ensiled in 1.0 liter anaerobic jars. After 45 days of ensiling, the results showed significantly lower

pH (3.7, 3.8, 4.1 and 4.5) and higher contents of lactic acid (10.5, 10.2, 9.3 and 7.3 %) respectively

for LAB+Enzyme, LAB, enzyme and control silage. While, the inoculation also effect the NDF%

(56.8, 60.8, 58.7 60.4) as well as in-vitro dry matter digestibility (IVDMD) (60.7, 58.3, 60.9 and

57.6) respectively, but have non-significant effect on CP and ADF contents. Concluded that

enzymes and lactic acid bacteria+enzymes mixture inoculant decreased neutral detergent fibre

content and increased in vitro dry and organic matter digestibility of silages.

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Khan et al. (2006c) examined the level of cane molasses on chemical composition and

fermentation characteristics of oat grass silage (OGS). And feeding value of oat grass silage (OGS)

ensiled with 2% molasses on intake, digestibility, milk yield and composition of Nali-Ravi

buffaloes (Bubalus bubalis). Oat grass (OG) harvested at 50-days of age was ensiled in laboratory

silos (transparent thick polyethylene bags of 10-kg capacity) with cane molasses at the rate of 0,

2, 4 and 6% of OG dry matter (DM). After 40 days of fermentation, silage pH was significantly

decreased (3.85, 3.63, 3.57 and 3.52) while lactic acid content increased (4.15, 4.43, 4.45 and 4.44)

with increasing level of cane molasses. However, a similar trends were observed for dry matter

(DM), crude protein (CP), neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents.

Two experimental diets of OG and OGS were formulated using 75:25 forage to concentrate ratio

on DM basis. Dry matter and CP intakes and its apparent digestibilities as well as milk yield were

similar in lactating buffaloes fed either OG- or OGS-based diets. Milk yield (4% FCM), milk

contents i.e. solids not fat and CP were similar while, milk fat, total solids and true protein content

were higher with OG compared with the OGS diet. The results of this study indicate that OG

ensiled with 2% molasses could safely replace 75% DM of green oat fodder in the diets of lactating

buffaloes.

Aragon et al. (2012) find out the effect of inoculation on nutrient content, fermentation,

and beef cattle performance for whole-plant corn silage treated with a commercial product BSM

(blend of Enterococcus faecium, Lactobacillus plantarum, and Lactobacillus brevis) was compared

to a control treatment with no silage additives (CT). Whole plant corn harvested at the milk/dough

stage of maturity (32.3% DM) chopped and treated with a silage inoculant (BSM) at the rate of 4

g/ton. BSM increased the fermentation rate with a significantly deeper pH (P < 0.01; 3.89 and 3.71

respectively, for CT and BSM), a significant increase in the total organic acids concentration (P <

0.05; 80.0 and 93.3g/kg DM), more lactic acid (P < 0.01; 50.3 and 61.4g/kg DM) compared to

CT. BSM-treated silage decreased DM by 3.0 % (P < 0.01) and had a higher digestible energy and

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a higher metabolizable energy concentration by 2.3 (P < 0.01) and 1.00 % (P < 0.05), respectively,

compared to untreated silage. The DM intake of silage treated with BSM increased by 6.14 %, and

improved weight gain and the feed conversion by 8.0 (P < 0.01) and 3.4%.

Iqbal et al. (2005) evaluated the effects of organic green culture (multiple probiotic) and

enzose (corn dextrose) on pH, lactic acid and chemical composition of mott grass silage (MGS).

Mott grass (MG) was harvested at 50 days of re-growth and ensiled in small laboratory silos with

organic green culture (OGC) and enzose at the rate of 0, 1.5 and 3 g kg/kg and 0, 1.5, and 3%

respectively. Silage pH decreased (4.7-4.1) and lactic acid (4.1- 4.5%) increased with increasing

level of enzose and remained unchanged by OGC treatment. The DM and CP recovery was higher

in inoculant (OGC) treated silages as compared to control (11.31 vs 11.1 and 10.6 vs 10.0

respectively). The NDF% of MGS increased (69.0-70.9) with increasing of OGC, however, was

unaffected by enzose treatment. While, ADF% of MGS was unaffected by varying level of enzose

and OGC. Concluded, that enzose can be used as a good source of fermentable carbohydrates for

preparing silage of fodders with low fermentable sugars.

Khadem et al. (2009) investigated the effects of barley flour on the fermentation

characteristics and its feeding value of alfalfa silage on the productivity of dairy cows. Alfalfa

forage was ensiled either with (12 kg: 100 kg dry matter bases) or without barley flour. Eighteen

multi-parous cows were assigned to three equal treatment groups using a completely randomized

design. Three isocaloric and isonitrogenous total mixed rations (TMRs) containing alfalfa hay,

ordinary alfalfa silage or barley flour mixed alfalfa silage were then prepared. The concentration

of lactic acid was higher in barley flour mixed alfalfa silage (510 g/kg DM) compared to ordinary

alfalfa silage (369 g/kg of DM). Although dry matter intake and milk production were not affected

by type of TMRs. It is concluded that the addition of barely flour when making alfalfa silage may

improve the fermentation process during ensilage and with non-significant effects on productivity.

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Saricicek. (2010) was investigate the effects of differ additives, molasses (M), dried poultry

mannure (DPM), microbial inoculant, sill-all 4x4 (SA), silofarm formiat dry (SD) and silofarm

liquid (SL) on the nutrient, silage fermentation and silage quality, energy content of corn silage.

Additives were applied at the rate of 3%, 30%, 10g/ton, 2.51/ton and 5kg/ton respectively. Results

showed that fleig score (quality) of silage were numerically higher in SA followed by M, control,

SL, SD and lower in DPM, are as 120.52, 119.50, 115.36, 110.64, 108.89 and 84.203 respectively.

Silage treated with DPM had significantly greater CP and CF content compared with other groups

(P<0.01). The pH value after 96 h of incubation was also higher in silage treated with DPM

compaired the other groups (P<0.01). The lactic acid concentration of silage were the highest for

silage with inoculant additive (P<0.01).

2.4. Feeding effects of cereal silages on production perfarmance of dairy animal

The feeding effects of cereals silages on growth performance, nutrients intake and

digestibility in dairy calves and the impact of feeding a replacement of maize fodder with maize

silage in total mixed ration (TMR) on nutrient intake, digestibility and production performance,

dry matter (DM), crude protein (CP), neutral detergent fiber (NDF) and acid detergent fiber (ADF)

intake, as well as digestibility and efficiency was varied in below published literature.

Charmley and Duynisveld. (2004) carried out an experiment to investigate the effects of

partially replacing silage with straw-barley-soybean meal mixtures on cow-calf performance.

Forty-eight multiparous cows were used in a 3 × 2 factorial experiment with three totally mixed

ration (TMR). The TMRs were formulated to contain 75, 50 or 25% silage (DM basis). The only

interactions between TMR and protein supplementation were a positive response to protein for

DM intake (P < 0.05) at the 50% silage level. Reducing the amount of silage in the TMR had no

effects on calf performance or milk production, except that milk protein concentration was higher

when the TMR contained 50% silage (quadratic effect; P < 0.04). However as the percentage of

silage in the TMR declined, cows lost less body weight (linear effect; P < 0.001) and appeared to

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improve in condition. It is concluded that silage can be successfully replaced with other ingredients

in winter beef rations, provided the nutrient concentration is balanced.

Bal et al. (1997) studied four stage of maturity (1. early dent, 2. quarter milkline, 3. two-

thirds milkline, and 4. black layer stages, with moisture contents of 69.9, 67.6, 64.9, and 58.0%

respectively) of whole-plant corn silage on intake, digestion, and milk production when fed as a

total mixed rations (TMR). TMRs containing 50% forage (67% corn silage and 33% alfalfa silage)

and 50% concentrate on dry matter basis were fed to twenty multiparous Holstein cows in a

replicated experiment with a 4 × 4 Latin square design with 28-d periods. Dry matter Intakes

(DMI) were similar across the four treatments and ranged from 3.73 to 3.79% of body weight.

Milk production was higher (33.4 kg/d) in group three and lowest (32.4 kg/d) in cows of group

1st. Milk protein production was also highest for group three (1.17 vs. 1.12 to 1.13 kg/d).

Apparent total tract digestion of dry matter, organic matter, crude protein and acid detergent fiber

was lowest for the cows of group four. Revealed, that two-thirds milk-line with some flexibility

between quarter and two-thirds milkline stages of maturity was optimum for corn silage.

Khan et al. (2006) conducted a trial to explore the impact of oat grass (OG) and oats grass

silage (OGS) based diets on feed intake, milk composition and milk production of lactating Nili-

Ravi buffaloes. Two tentative diets of oat grass and oat grass silage were formulated using 25:75

concentrate to forage ratio on dry matter (DM) basis. Dry matter (DM) and crud protein (CP)

intakes were similar among the treatments. While, neutral detergent fiber (NDF) intake was greater

in OG diet as compared to OGS diet fed to buffaloes (8.35 and 7.16 kg/day respectively). Whereas,

apparent DM, CP and NDF digestibility and milk yield were also non-significant between the

treatments. Milk fat, total solids and true protein (6.87 vs 5.95, 16.0 vs 15.1 and 3.35 vs 2.92 %

respectively) contents in buffalo milk were higher with OG diet compared with OGS diet. Solid

not- fat, crude protein and non-protein N contents were similar in buffalo milk fed either OG or

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OGS based diets. Concluded that OGS could safely replace 75% DM of green oat fodder in the

diets of lactating buffaloes without effecting intake, digestibility and milk yield.

Caplis et al. (2005) evaluated the effects of silage, concentrates and combination of both

in winter finishing of beef steers. All the groups’ were assigned to 6 treatments randomly having

18 animals in each group. Feeding treatments were (1) silage only, fed ad lib (SO), (2) silage and

low level of concentrate fed separately (LS), (3) silage + concentrates offered as TMR (LM), (4)

silage + medium level of concentrates fed separately (MS), (5) silage + medium level of

concentrates fed as TMR (MM), (6) restricted silage + concentrates ad libitum (AL). Silage dry

matter intake were decreased (P < 0.001) significantly, however, total dry matter intake of

individual animal was increased significantly (P < 0.001) with the increase in concentrate levels.

The weight gain was found higher in the group fed AL followed by MM, MS, LM, LS and SO,

respectively. Feeding method had no interaction with concentrates level. Silage feeding as TMR

had increased dry matter intake significant (P < 0.001) as compared to feeding separately.

Moreira et al (2006) studied the impact of alfalfa silage (AS): corn silage ratio in 50:50

forage: concentrate. Treatments were assigned randomly among the groups to evaluate the effect

of diet on production and rumen traits of dairy animals. Dry matter intake (DMI) was significantly

increased (P < 0.001) in the group ranges 23.4 with 100% corn silage to 24.7 and 24.3 kg/d with

the inclusion of alfalfa silage in the diet. Milk production followed the similar trend to dry matter

intake having 39.4 kg/d with 100% corn silage against 40.6 kg/d with corn silage: alfalfa silage

mixture. Chemical composition of milk yield had decreased fat, solid not fat and lactose contents

significantly (P < 0.004) whereas there was increased protein contents with the increase in corn

silage in the diet.

Ahvenjarvi et al. (2006) replaced gradually grass silage with whole crop barely silage and

its effect on feed intake, milk production and total tract digestibility in lactating Holstein cows.

Four rumen cannulated dairy cows in early lactation, were assigned to four experimental diets for

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a periods 21 days. The diets (forage part) comprised a mixtures of grass silage and whole crop

barley silage supplemented with 8.9 kg/d of concentrates (DM basis). The percentage (%) of barley

silage in the forage was 0 (control), 20, 40, and 60 of DM. Results showed that replacement of

grass silage with barley silage had no effect on DM, digestible organic matter, or neutral detergent

fiber (NDF) intake. Whereas, digestible NDF (dNDF) intake (7.34, 6.90, 6.74 and 5.91 kg/day)

respectively decreased with increasing the percentage of barley silage. Increases in the proportion

of barley silage linearly decreased milk yield (33.1, 33.2, 32.8 and 31.0 kg/day respectively).

Decreases in milk yield due to inclusion of barley silage were attributed to decreases in diet

digestibility and nutrient supply to the animal. Decreases in organic matter and NDF digestibility

were attributed to the higher indigestible NDF concentration of barley silage compared with that

of grass silage and to the smaller pool size of dNDF in the rumen.

Tauqir et al (2007) a feeding trial was conducted in thirty early-lactating (45±4 days),

multi-parous Nili buffaloes (Bubalus bubalis), ten in each group, were allotted to three

experimental TMRs. To evaluate the feeding worth of jambo grass (Sorghum bicolour×Sorghum

sudanefe) silage and mott grass (Pennisetum purpureum) silage as a replacement of conventional

fodder (jambo grass) in the diet of dairy buffaloes. The control (JG) TMR contained 75% jambo

grass while the other two TMR contained 75% jambo grass silage (JGS) and 75% mott grass silage

(MGS) of dry matter (DM). The remaining 25% DM in each diet was supplied by concentrates,

forage and concentrates were mixed daily and fed ad libitum for 120 days to the experimental

animals. Dry matter intake (DMI) was higher in control (13.3 kg/day) diet compared with JGS

(12.5 kg/day) and MGS (12.03 kg/day) diets. However, DMI as % body weight did not differ

significantly among the treatments. Apparent total tract digestibilities of DM, CP and NDF were

similar in buffaloes fed JG, JGS and MGS diets. However, milk yield and composition were also

similar in buffaloes fed JG and silage based diets. The results indicated that jambo grass and mott

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grass ensiled with 2% molasses for 30 days could safely replace the conventional fresh grass fodder

(75% DM) in the diet of lactating Nili buffaloes without affecting their milk yield.

Bilal (2008) carried out an experiment to determine the effect of feeding mott grass (MG),

mott silage (MGS) and their combinations on the production performance of lactating Sahiwal

cows. Numerically maximum (10.08 kg/day) dry matter intake (DMI) was observed in cow fed on

MG supplemented with molasses and minimum (9.46 kg/day) in those fed MGS ensiled with

molasses. Dry matter intake as a percent body weight ranged from 2.69 to 2.80. However, daily

CP intake varied from 1.21 to 1.30 kg/day, variation in CP intake was attributed to variation in

DMI and NDF intake ranged from 7.08 to 7.60 kg were also non-significant. Milk yield (4% FCM)

ranged from 7.84 to 9.06 Lt/day. Maximum FCM yield was in cows fed mott grass/silage in

combination and minimum in those cows fed mott silage in which no additive was used.

Statistically, difference in milk yield was non-significant (P>0.05) in cows fed mott grass alone

and mott silage alone. Milk composition of cows fed experimental diets remained unaltered.

Significantly higher in-vivo dry matter digestibility (62.20-62.84%) was found in cows fed mott

grass/silage in combination and minimum (58%) in cows fed silage in which no additive was used.

While, non-significant difference among digestibilities of mott grass and silage based diets.

Concluded that MGS is the best substitute of MG fodder, mott grass silage alone or in combination

can be used in dairy animals without any negative impact on dry matter intake, milk production,

milk composition and digestibility.

Cook et al (2008) conducted an experiment to investigate the impact of ground corn (GC)

or steam flaked corn (SFC) feeding based diets on either annual ryegrass silage (RS) or a

combination of 50, 50% blend of RS and corn silage. Diets were fed as TMR. Significant

difference (P < 0.05) were found in DM (23.55 and 23.04 vs 19.65 and 19.63 kg/day), OM (20.05

and 18.78 vs 16.02 and 16.59 kg/day) and NDF (7.08 and 6.66 vs 6.02 and 5.94 kg/day) in group

fed blend (CS) as compared to cows fed annual ryegrass silage (RS) for GC and SFC respectively.

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Whereas total tract digestibility percent of OM (69.45 79.95 vs 67.44 70.08), NDF (50.47 55.74

vs 47.85 44.09) and ADF (47.87 55.85 vs 39.02 34.73) were higher in group fed ryegrass silage as

compared to blend. Cows fed Blend diet had increased (P < 0.05) energy corrected milk as

compared to ryegrass silage and resulted in improved milk efficiency (1.30 and 1.53 vs 1.46 and

1.67) in terms of kg of milk per kg of DMI. Concluded that feeding a blend of annual ryegrass and

corn silage is more desirable than feeding diets based on RS as the sole forage. Supplementing

diets with SFC improved performance and efficiency compared with GC across forage sources.

Vargas-Bello-Pérez et al. (2008) conducted a feeding trial to evaluate the effects of soybean silage

(SS) and 4th cut alfalfa silage (AS) as a component of total mixed ration (TMR) on intake and yield

of lactating Holstein cows. Tow iso-nitrogenous TMR were formulated with 48:52 forage

concentrate ratio. The forage part of the TMR consisted of 72% soybean or alfalfa silage while,

the remaining 28% comes from corn silage in both diets. Significantly higher DMI (25.1 vs 23.5

kg/d) and milk yield (35.5 vs 37.2 kg/day) were observed in the group fed alfalfa silage compared

with soybean silage. Statistically no difference were observed in energy corrected milk and its

efficiency in both the feeding groups of cows. There were no significant difference observed by

the dietary treatments in fat, protein, SNF and lactose contents of milk.

Abrahamse et al. (2008) revealed the feeding value of five different total mixed rations

(TMR) on intake and milk production in early and late lactating dairy cows. The treatments were

as: i) TMR with 55% roughage (27.5% corn silage and 27.5% grass silage) and 45% concentrate

(50:50 mixture of a concentrate rich in structural and a concentrate rich in nonstructural

carbohydrates; treatment CON), ii) TMR with 55% corn silage and 45% of the concentrate mixture

(treatment RCS), iii) TMR with 55% grass silage and 45% of the concentrate mixture (treatment

RGS), iv) TMR with 55% of the roughage mixture and 45% of the concentrate rich in

nonstructural carbohydrates (treatment CNS), and v) TMR with 55% of the roughage mixture and

45% of the concentrate rich in structural carbohydrates (treatment CSC). The DMI was lower (P

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< 0.001) in RGS (TMR) compared with the other treatments (17.3, 20.1, 19.5, 19.5, 19.7 kg/day).

Milk production did not differ between treatments. While, milk fat content was lesser (P<0.001)

in diet RCS (3.55 vs 4.10 %) and greater milk protein content (3.53 vs 3.45 %) than diet RGS.

Intake rate increased when the amount of grass silage decreased, these effects were in line with a

decreased DMI of the RGS diet vs, the other treatments, probably related to the high neutral

detergent fiber (NDF) content. Lactation stage did affect short-term feed intake behavior and DMI,

although different grass silages were fed during early and late lactation. The results indicate that

short-term feed intake behavior is related to DMI and therefore may be a helpful tool in optimizing

DMI and milk production in high-production dairy cows.

Cook et al. (2009) a feeding experiment was conducted to determine the effects of feeding

different proportions of corn silage and ryegrass silage in a total mixed ration (TMR) on the

performance of lactating Holstein dairy cows. Forage provided 49% of the dietary dry matter while

remaining supplemented with ground corn (GC), steam-flaked corn (SFC), and hominy feed (HF)

according to NRC requirement. The proportions of ryegrass silage in TMR, 100, 75, 50, or 25%

of the total forage (49%) dry matter, with corn silage supplying the remainder. The results indicates

that dry matter intake (22.1, 21.7, 21.7 and 20.7 kg/day respectively) and milk protein percentage

(3.00, 2.92, 2.85 and 2.85 respectively) decreased linearly with increasing proportions of ryegrass

silage, but milk protein yield was similar among forage treatments. While, non-significant

difference in milk yield, milk fat percentage and yield, and energy-corrected milk yield were

observed among forage treatments. However, total tract digestibility percentage of NDF (P = 0.03;

41.8 42.4 48.2 48.1) and ADF (P = 0.006; 39.2 41.3 48.2 45.6) increased linearly with increased

ryegrass silage in the diet, but non-significant effects on DM ,CP and OM digestibility .Concluded

that feeding of corn silage and ryegrass silage in combination is more desirable than feeding

ryegrass silage alone.

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Tauqir et al. (2009) a feeding trial was conducted on lactating Nili Ravi buffaloes (Bubalus

bubalis) to find out the effects of Jambo grass (Sorghum bicolour Sorghum sudanefe) silage as a

replacement of conventional fodder (Jambo grass; JG). Two iso-nitrogenous and iso-energetic

experimental total mixed ration (TMR) were formulated with 75% of forage (JG or JGS) and 25%

concentrate on DM basis. Twenty early lactating dairy buffaloes were divided into two equal

groups and fed experimental diets for 60 days at ad libitum. Daily dry matter intakes (p<0.05; 13.3

vs 12.5 kg/day), CP (1.74 vs 1.63 kg/day), neutral detergent fiber (8.35 vs 7.32 kg/day) and acid

detergent fiber (4.31 vs 4.07 kg/day) were significantly higher in animals fed control TMR than

those fed Jambo grass silage TMR. A significant difference of nutrients intakes among JG (control)

and JGS, TMR may be due to the presence of fermentation products in ensiled material that might

have depressed the intake in silage based TMR. Apparent nutrients digestibilities and milk yield

were non-significantly between treatments. The present results indicated that JG ensiled with 2%

molasses for 30 days could safely replace the conventional fresh Jumbo grass fodder in the diet of

lactating Nili Ravi buffaloes without affecting their milk yield.

Hayashi et al. (2009) summarized the effects of whole crop maize silage (MS) as a substitute

for rice straw (RS) on feed intake and milk production in mid-late lactating Murrah buffalo and

Jersey-Hariana cattle in Nepal. Experimental animals of each species were fed the RS (ad libitum)

as basal diet and supplemented with 0.68% concentrates of body weight on DM basis. Maize silage was

substituted at 0, 33%, 67% and 100% in group T1, T2, T3 and T4, respectively for rice. The DMI was

significantly higher (P<0.05) in T3 followed T4, T2 and lowest in T1 treatment, in both species

(10.95, 9.69, 9.84, 8.66 kg/day and 9.17, 9.0, 8.31, 8.10 kg/day for Buffalo and cattle respectively).

The substitution of MS for RS increased the crude protein intake and the TDN intake in the both

species. Whereas, the buffalo showed the highest milk yield feed T4 followed by T3, T2 and T1

(2.82, 2.64, 2.60 and 2.31 kg/day respectively), but the cattle showed non-significant difference in

Review of Literature

25

their milk yield among the treatments. Concluded, that substitution of MS for RS increased the

feed intake in both species and milk production in buffalo.

Tanaka et al. (2010) investigated the effect of citrus pulp silage on production performances in

Holstein lactating cows for a period of two weeks. Citrus pulp silage were fed to experimental animals as

TMR having 20% citrus pulp silage on DM basis. Results of the experiment suggested that there was non-

significant difference (P>0.05) on DMI, milk production and its components were observed as compared

to control group.

Oshita et al. (2007) evaluated the effects of maturity stages i.e. early dent and late dough

stage of corn silage (hybrids) on milk performance of Holstein cows in the northern part of Japan.

The stage of maturity for the early-maturing (EH; 80 d relative maturity) and the mid maturing

(MH; 93 d relative day) were early dent and late dough stage, respectively. The plant and silage

yields for MH were higher than those for EH (70.68 vs 57.57 and 65.24 vs 56.61 ton/ha,

respectively). Therefore, the DM yields of prepared silage per area were similar for both

treatments. For a feeding trial twelve multiparous mid-lactation Holstein cows (58±13 days in

milk) were fed diets based on EH or MH corn silage for 3-week periods. Results showed that DM

intake for EH was found to be higher (p<0.05) than that for MH silage (10.0 vs. 9.1 kg/day).

However, milk production and milk composition and feed efficiency per total feed intake were

similar among the treatments. Concluded that differences in maturation in corn hybrids affect the

silage intake of dairy cows. It may be advantageous to plant early hybrid corn with a reduction in

purchased feed costs for dairy cows under the climatic conditions of the northern part of Japan.

Oltjen and Bolsen. (1980) compared the nutritive value of winter wheat, barley and spring

oats, corn silages, fed to growing steers in three different feeding experiments. Corn and small

cereals forages were harvested at hard-dent stage and dough stage respectively, except for an

additional milk-stage harvest of Eagle wheat in trial 2. Diets were formulated as silage (86%) and

supplement (10% of a milo-SBM mix and 4%) on dry matter basis. In trial 1st, significantly higher

Review of Literature

26

DMI was observed in steers fed corn silage followed by barley, wheat (eagle, Arthur and blue boy

II) respectively, (8.75, 8.15, 7.40, 6.81 and 6.71 kg/day respectively). However, significantly

(P<0.05) a similar pattern was also observed for daily weight gained (1.28, 1.18, .87, 87 and .71

kg). Although efficiency of gain was similar to that for steers fed barley silage (P>.05). Wheat

silages supported poorer performance than corn or barley silages; steers fed Blue Boy II wheat

gained slowest (P<0.05). Concluded that dry matter intake was affected more by composition of

the silage than by plant species, but efficiency of gain was influenced more by plant species.

Regression model analysis revealed that silage dry matter and acid detergent fiber were

consistently responsible for daily gain and dry matter intake responses. Each additional percentage

unit of silage dry matter content increased daily dry matter consumption by .08 kg/steer; each

additional percentage unit of silage acid detergent fiber decreased daily gain .04 kilogram.

Statement of problem

Low milk production and growth rate of Nili-Ravi Buffalo in Pakistan is mainly due to

animals consuming a low nutritional diet and fluctuation in the supply of forage for livestock

feeding. Including, in the country there are two obvious fodder scarcity periods (extremes of

summer and winter) but during rest of the year fodder availability is fairly regular and abundant.

The preservation of this abundance fodder by silage, at its proper stage of maturity, in feasible

storage technology can solve the problem.

Objectives:

1. To investigate factors affecting the nutritional quality of silage

2. Effect of stage of cutting and types of silo on chemical composition i.e. dry matter, crude

protein, NDF and ADF contents of silage

3. To evaluate different level of additive (Sil-all) to enhance silage quality

4. Effect of silage feeding on growth and production performance in Nili-Ravi buffaloes

Review of Literature

27

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33

CHAPTER 3

EXPERIMENT-1

Impact of flowering stage (maturity) on fermentation qualities, digestibility and nutritive

value of silages made from maize, sorghum and oats fodders

ABSTRACT

The objective of current study was to investigate the effects of flowering phase (maturity) i.e. early

bloom (20% flowering), mid bloom (50% flowering) and full bloom (100% flowering) on

chemical and physical characteristics of silages, made from maize, sorghum and oat cereal fodders

in grown in subtropical conditions. The three fodders were harvested at three different maturity

stages, chopped and ensiled in laboratory silos for 30 days fermentation period. The analysis of

variance revealed that the increase in maturity from early to full bloom significantly (P < 0.05)

increased dry matter (DM %) in maize (18.82±0.02 to 25.80±0.05), sorghum (20.65±0.01 to

29.47±0.01) and oat (17.68±0.01 to 26.54±0.01) silages, and decreased crude protein in all. The

NDF and ADF increased with increase in maturity for sorghum and oats silage, while it decreased

for maize silage. Lactic acid increased linearly (P < 0.05) with increasing maturity while pH, ME

and in vitro dry matter digestibility (IVDMD) decreased. Stage of maturity also had significant

effect on physical characteristics (color, smell and structure) of silage in all fodder crops. The

highest flieg score was recorded in full bloom stage of maturity (100% flowering) due to low pH

and higher DM recovery in all cereals silages. It was concluded from the current study that quality

silages can successfully be made from cereal fodders grown in subtropical conditions.

Experiment 1

34

INTRODUCTION

The shortage of good quality fodder, round the year, is the major constraint for livestock

production in Pakistan (Khan et al., 2011). The seasonal fodder production poses a great challenge

for livestock farmers to feed their animals when fodder supply is limited especially during summer

(May - July) and winter (November – January) months (Rasool et al., 1996). The conservation of

fodder in the form of silage is a viable solution to ensure its supply during those lean periods (Khan

et al., 2011).

The quality of silage is dependent on the availability of fermentable substrates (McDonald

et al., 1981) energy density, and water content in plants (Bal et al., 1997). As the plant matures,

the water-soluble carbohydrates decrease, thereby decreasing the fermentation activity of bacteria

(Jhonson et al. 2003). Too early or too late harvesting stage not only impairs the energy density of

whole plant but also affects the optimum moisture level required for good silage preservation (Bal

et al., 1997). Therefore, optimum stage of maturity is important to harvest maximum nutrients for

livestock feeding.

Optimal harvesting stage to increase yield and silage quality of maize, in literature, varied

from tasseling stage (Fu et al., 2011), one-third milk line (Johnson et al., 2002), late dough stage

(Vecchiettini et al., 2003) to two-thirds milk line stage (Fariani et al., 1994). When sorghum was

harvested at late-milk, late-dough and hard-grain stages of maturity, higher nutritive values were

noticed at late milk stage silages (Sonon & Bolsen, 1996). Also, Khan et al. (2012) reported that

advancing development (maturity) of corn from 30.0 to 42.0% (i.e. black layer stage) dry matter

(DM) at ensiling did not affect DM intake, milk production and composition in dairy animals.

Despite the difference in optimal harvesting stage for silages, it is also worth mentioning that there

are even very few studies investing the effects of maturity stage on silage quality in subtropical

environment as in Pakistan. Under such scenario, the studies are needed to tackles the issue of

silage production for maximum feed supply to the livestock.

Experiment 1

35

The objective of current study was to examine the effect of varying maturity stages of three

different fodders (maize, sorghum and oats) on fermentation characteristics and nutritive value of

silages under local environmental conditions.

MATERIALS AND METHODS

Sowing and harvesting of cereal fodder crops

The three fodder crops i.e. maize (Zee maize), sorghum (Sorghum bicolor) and oats (Avena

sativa) were used for silage making. The sorghum, maize, and oats were planted during the month

of June, July and November, respectively on agriculture field of Dairy Animals Training and

Research Center, University of Veterinary and Animal Sciences, Ravi Campus Pattoki, Pakistan

(31°1'0" North, 73°50'60" East, 186 meters elevation). Each fodder crop was harvested at three

different stages of maturity. The maturity stages were based on the flowering in the field and

categorized as; 1) 20 % flowering (20% of plants in the field had shown flowers), 2) 50%

flowering, 3) 100 % flowering. The detail of planting and harvesting at different maturity stages

has been presented in Table 3.1. At each harvesting time, the respective fodder was randomly cut

during a clear day from four different parts of the field and chopped by mechanical chopper

(Fimax, V-Belt Driven, MC10X, Turkey) with a chop size of about 2 cm.

Ensiling of chopped fodder in bag silos

The chopped forage taken from different parts of the field was mixed to make a

representative sample for silage making. The mixed sample was packed in laboratory silos

(transparent thick polyethylene bags) with capacity of 40 kg having dimensions 80×40 cm. The

chopped materials were packed in the bag silo step by step. To fill the bag, about 3-6 kg of chopped

forage was placed in the bag each time and pressed manually for compaction to remove air. The

procedure was repeated till the bag was full and then sealed immediately. All the bags were labeled

and stored under shed at room temperature for fermentation.

Experiment 1

36

Physical quality evaluation of silage

After 30 days of fermentation period, the bags were opened and samples were taken for

physical and chemical analysis. For physical analysis, the quality of silages was determined by

color, smell, and structure along with total flieg score described by Kilic (1986). For color

evaluation, the scale 1- 4 was used on the basis of change in green color from dark brown , dark

green to pale yellow; for smell , the scale 1-7 was used on the basis of repugnant putrid smell to

acidic sweet pleasant smell; for structure, the scale 1-4 was used on the basis of softness of leaves

and stem as well as its ability to remain intact after squeezing the silage tightly in hand and then

opening from breaking into small pieces to break into two or three pieces. The same person scored

the silages for smell, color and structure to avoid any bias. Flieg score was calculated using a

formula (flieg Score = 220 + (2 x Dry Matter% - 15) - 40 x pH) reported by Kilics (1986). The

flieg score with value 81-100, 61-80, 41-60, 21-40 and 0-20 represented the silage quality a very

good, good, medium, low and poor, respectively. The overall silage quality was classified into

categories as poor, medium, good and very good on the basis of cumulative score obtained from

color, smell, structure and flieg score.

Chemical quality and pH analyses of silages

Chemical analyses were done to determine pH, lactic acid, dry matter content (DM), crude

protein (CP), neutral detergent fiber (NDF), and acid detergent fiber (ADF). Approximately 25g

sample was taken from each bag immediately after opening. The sample silage was adulterated

with 100 ml of distilled water (Hart and Horn., 1987). After hydration for 10 min using blender,

the diluted material was then filtered through cheese cloth and then pH was determined by using

a digital pH meter. The liquid obtained was further filtrated through Whatman 54 filter paper,

centrifuged and kept at 20 ̊ C for lactic acid determination by high pressure liquid chromatography

(Muck and Dickerson., 1988). Also approximately, 250g sample (in triplicate) was taken from

each bag, dried in a hot-air oven (Memmert, Beschickung-Loading Model 100-800, Germany) at

Experiment 1

37

60°C for 72 hours (for DM%), then ground through hammer mill (Wiley laboratory Mill, Standard

Model No. 2, Arthur H. Thomas Company, USA) making particle size of about 0.5 to1mm and

stored in pre labeled bottles for further laboratory analyses. Nitrogen (N) contents of samples were

determined by procedure AOAC. (1990) using Kjeldahl apparatus (ID 984.13), and then

multiplying the N concentration by a factor 6.25 to calculate CP. The NDF and ADF contents were

determined according to Van Soest et al. (1991). The gross energy of the silage samples was

determined through the IKA C-2000 Bomb Calorimeter, while metabolizable energy (ME) was

calculated as 63% of the gross energy (Mandal et al., 2003).

In vitro dry matter digestibility of silages

The in vitro dry matter digestibility (IVDMD) trials were conducted at University of

Sydney, Camden. The dried samples were taken from Pakistan to Camden by air cargo. For

IVDMD study, rumen liquor (inoculant) was collected from rumen of cannulated lactating

Holstein cows managed on pasture and cereal-based concentrate (9kg DM/cow/day), at

Corstorphine farm, University of Sydney. The collected rumen liquor was filtered through various

layers of cheese cloth and mixed with buffered minerals solution in 1:2 ratio and placed at 39 ̊ C

under O2 free environment. Dry matter digestibility (DMD) was determined in vitro by batch

incubation of samples in rumen liquor (Wang et al., 1999). All the dried samples from respective

cereal silages were incubated in duplicate using ANKOM filter bags technology (New York,

USA). The open side of the bag (having 0.5g ground sample) was sealed with heat sealer impulse,

and then put into a 50ml dark bottle. All the bottles were injected with 25ml of buffer solution

under anaerobic condition then sealed with rubber lid. Bottles including blanks (inoculums only)

were incubated in rotary incubator for 48hrs at 39 ̊ C and rotated 90 times per minute. After 48 h

of incubation the bags having digested sample were removed from the flasks, washed under

running tap water then dried in oven at 60 ̊ C for 48 hours. The IVDMD% was calculated from the

Experiment 1

38

difference of the dry weight of sample and residues remained in the bag after 48 h of digestion

divided by weight of sample×100 (Wang et al., 1999).

Statistical Analysis

The data were analyzed by analysis of variance, using General Linear Models procedures

of SAS (SAS 9.1.3). Differences of means among main effects were compared by Fisher’s least

significant difference test (Steel et al., 1997).

RESULTS

Chemical Composition and Nutritive Value

Chemical composition of three silages (maize, sorghum and oats) harvested at three

different stages of maturity i.e. early bloom (20% flowering), mid bloom (50% flowering) and full

bloom (100% flowering) has been presented in Table 3.2. DM content (%) increased with

increasing maturity from early to full bloom (P < 0.05) in maize (18.82±0.02 to 25.80±0.05),

sorghum (20.65±0.01 to 29.47±0.01) and oats (17.68±0.01 to 26.54±0.01) silages respectively, but

in contrast CP and ME contents were significantly (P<0.05) decreased in all cereals silages with

increasing maturity (Table 3.2). However, with advance maturity increasing pattern of NDF and

ADF were also observed in sorghum and oats silage with progressed age form early to full bloom

stage, but in contrary tended to be decreased NDF and ADF in maize silage with advancing

maturity (Table 3.2).

Fermentation characteristics and IVDMD of silages

The results showed that with the increase in maturity stage of silages, fermentation

characteristics significantly got better (Table 3.3). The pH with advancing maturity linearly

decreased in maize 4.29±0.005, 4.24±0.005 and 3.94±0.008, sorghum 3.95±0.01,3.83±0.008 and

3.62±0.017 and oats silage 4.04±0.008, 3.95±0.02 and 3.71±0.008 from early to full bloom stage,

respectively (P < 0.05, Table 3.3). Whereas, lactic acid concentration significantly (p<0.05)

Experiment 1

39

increased with advance maturity. However, IVDMD decreased with increasing maturity for

sorghum and oats but not or maize (P < 0.05, Table 3.3).

Physical Quality of Silages

Effect of maturity on physical characteristics including color, and smell structure and flieg

score has been shown in Table 3.4. Maturity had non-significant (P>0.05) effect on color of the

silages in all cereals. However, the score for smell and structure positively increased with the

increase in maturity for maize, sorghum and oat silages from early bloom to full bloom (P<0.05).

Numerically higher flieg score was observed in full bloom followed by mid bloom and early bloom

stage of maturity, respectively, for all silages. The cumulative effect of all physical traits indicated

that silages had highest quality at 100 % maturity stage of flowering (full bloom) in all cereal

silages.

DISCUSSION

Chemical Composition and Nutritive Value

The finding of increased DM and decreased CP, and ME with increasing maturity in our

study was consistent with previous studies (Khorasani et al., 1997; Khan et al., 2011). The current

results are in agreement to Khan et al. (2011) who reported that NDF and ADF in maize silage

decreased with increasing maturity while it increased in other cereal silages like sorghum and

millets. The increased DM and lower CP and ME with increasing age could be attributed to

increased lignifications and decreased content of leafy part of plant. The decreased NDF and ADF

in maize silage with increasing maturity could be due to the deposition of starch into grains.

Similarly, Khan et al. (2012) who found that difference in maturity at harvest during grain filling

had a major effect on the carbohydrate structure (starch:NDF ratio) and fatty acid (FA) content of

corn silages.

Experiment 1

40

Fermentation characteristics and IVDMD

The linear decrease in pH values of maize, sorghum and oats silages from early to full

bloom stage of maturity was in agreement with the findings of Sarwatt et al. (1989) who described

that pH values of maize silages decreased with advancing growth of fodder. Similarly, Khan et al.

(2011) also reported that ensiled fodder (maize, sorghum and millet) at initial stage of growth did

not decrease pH quickly. The decrease in pH is mainly due to the accumulation of lactic acid as a

result of fermentation. Lactic acid contents in the our study were supported by the findings of Khan

et al. (2011) who ensiled maize, sorghum and millet fodder at pre-heading, heading and milk stage

of maturity and concluded that lactic acid contents increased with the advancement of age of

fodder. Similarly, Bal et al. (1997) and Harrison et al. (1998) reported that maize silage harvested

at milk stage of maturity have highest concentration of lactic acid. The pre-requisite for the

development of the lactic acid bacteria during the early stages of ensilage are the contents derived

from the plant juices (water soluble carbohydrates) released by plasmolysis as a result of plan cell

wall breakdown (McDonald. 1981). Khan et al. (2011) reported that ensiled fodder (maize,

sorghum and millet) at initial stage of growth had low level of available water soluble

carbohydrates (WSC). Bergen et al. (1991) reported that WSC was greater in silages made from

barley, wheat and oats fodders harvested at milk stages. The range of pH values in current study

ranging between 3.62 and 3.94 at full stage of maturity (100% flowering) in all cereals silages

were consistent with the reports of McCullough (1978), who observed that pH value less than 4.2

was indicative of good quality preserved silage.

The decreased IVDMD in sorghum and oats silages, but unchanged in maize silage with

advancing maturity was supported by Edmisten et al. (1998) who reported that IVDMD of small

grain cereals (barley, oat, rye, and wheat) silages harvested at six growth stages, decreased from

vegetative to the milk stage and then remained similar or declined marginally to the hard dough

stage. The decline in IVDMD from growing to the boot stage was probably due to the consistent

Experiment 1

41

small increase in lignification of stems. Russell et al. (1992) also investigated the effect of growth

on IVDMD harvested at 0, 14 and 28 days intervals after physical maturity and suggested that later

harvest did not affect IVDMD of the maize forages.

Effect of maturity proceeding to physical quality of cereals silages

The improvement in physical traits of silages with increasing age was in agreement to

previous studies (Khan et al., 2011; Khan et al., 2012). Higher values for color, smell, structure

and flieg score in full bloom followed by mid bloom and lower in early bloom stage of maturity

could be due to low pH and higher DM as well as higher lactic acid concentration across all crops

silages in advanced maturity stage. Türemiş et al. (1997) reported that low pH and acetic acid

contents resulted in high physical quality scores.

Conclusion

It was concluded from the current study that quality silages can successfully be made from

cereal fodders grown in subtropical conditions.

Experiment 1

42

LITERATURE CITED

AOAC (Association of Official Analytical Chemists). 1990. Official Methods of Analysis. 15th

Ed., Association of Official Analytical Chemists, Arlington, Virginia, USA.

Bal MA, Coors JG, Shaver RD .1997. Impact of the maturity of corn for use as silage in the diets

of dairy cows on intake, digestion, and milk production. J Dairy Sci. 80: 2497–2503.

Bergen WG, Byrem TM, Grant AL. 1991. Ensiling characteristics of whole-crop small grains

harvested at milk and dough stages. J Anim Sci. 69: 1766-1774.

Edmisten KL, Green JT, Mueller JP, Burns JC. 1998. Winter annual small grain forage potential.

ii. Quantification of nutritive characteristics of four small grain species at six growth stages.

Commun. Soil Sci. Plant Anal., 29 (7&8), 881-899.

Fu F, Guo C, Tang Q, Liu J, Li W. 2011. Growth dynamics and optimal harvesting stage of two

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Fariani A, Warly L, Matsui TA, Fujihara T, Harumoto T. 1994. Rumen degradability of Italian

Ryegrass (Lolium multiflorum, L) harvested at three different growth stages in sheep.

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Harrison JH, Johson, Riley LR, Xu S, Loney K, Hunt CW, Sapienza D. 1998. Effect of harvest

maturity of whole plant corn silage on milk production and component yield and passage

of corn grain and starch into feces. J. Dairy Sci. 79(1):149.

Hart SP, Horn FP. 1987. Ensiling characteristics and digestibility of combinations of turnips and

wheat straw. J. Anim. Sci., 14: 1790-1800

Khan NA, Tewoldebrhan TA, Zom RLG, Cone JW, Hendriks WH. 2012. Effect of corn silage

harvest maturity and concentrate type on milk fatty acid composition of dairy cows. J.

Dairy Sci. 95:1472–1483.

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Khan SH, Azim A, Sarwar M, Khan AG. 2011. Effect of maturity on comparative nutritive value

and Fermentation characteristic of maize, Sorghum and millet silages. Pak. J. Bot., 43(6):

2967-2970.

Khorasani GR, Jedel PE, Helm JH, Kennelly JJ. 1997. Influence of stage of maturity on yield

components and chemical composition of cereal grain silages. J. Anim. Sci. (77): 259-267.

Kilic A. 1986. Silo feed (Instruction, Education and Application Proposals). Bilgehan Press, Izmir,

pp: 327.

Mandal AB, Paual SS, Pathak NN. 2003. Nutrient requirements and feeding of buffaloes and cattle.

Published by Int. Book Distributing Co. Charbagh, Lucknow, India. P-23.

Rasool S, G. Ahmad G, Abdullah M. 1996. Fodder conservation, crop residues and by products in

livestock production system. Proceeding of national conference on the improvement

production and utilization of fodder crops in Pakistan. March 21-27, Pakistan Agriculture

Research Coubcil / FAO, Islamabad, Pakistan, p. 176-186.

Russell JR, Irlbeck NA, Hallauer AR, Buxton DR. 1992. Nutritive value and ensiling

characteristics of maize herbage as influenced by agronomic factors. Anim. Feed Sci.

Technol. 458 38, 11–24.

Steel GD, Torrie JH, Dickey DA. 1997. Principles and procedures of statistics, 3rd ed., Mc Graw-

Hill, New York.

Sonon RN, Bolsen KK. 1996. Effects of cultivar and stage of maturity on agronomic

characteristics, chemical composition and nutritive value of forage sorghum silages. Adv.

Agric. Res. 5(3):1-17.

Sarwatt SV, Mussa MA, Kategile JA. 1989. The nutritive value of ensiled forages cut at three

stages of growth. Anim. Feed Sci. Technol. 22: 237-245.

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Johnson LM, Harrison JH, Davidson D, Mahanna WC, Shinners K, Linder D. 2002. Corn silage

management: Effects of maturity, inoculation and mechanical processing on pack density

and aerobic stability. J. Dairy Sci. 85(2): 434-444.

Muck RE, Dickerson JT. 1988. Storage temperature effects on proteolysis in alfalfa silage. Trans.

ASAE. 31:1005-1009.

McDonald P. 1981. The biochemistry of silage. John Wiley and Sons, Ltd. Chichester, United

Kingdom.

McCullough ME. 1978. Silage-some general consideration. Pages 1-26 in fermentation of silage-

A review. (Ed.):M.E. McCullough. National feed ingredients Assoc., West DesMoines, IA.

Türemiş A, Kizilşimşek M, Kizil S, İnal İ, Sağlamtimur T. 1997. Farklı katkı maddelerinin

çukurova koşullarında yetiştirilen bazı yazlık yem bitkisi ve karışımlarından yapılan

silajlar üzerine etkisi. Türkiye I.Silaj Kong. Bildiri Kitabı. Bursa, s.166-175.

Vecchiettini M, Cinti F, Sandrini E. 2003. Effect of harvest stage on maize silage production. In:

Kirilov A, Todorov N, Katerov I. (Eds.), optimal forage production for animal. Production

and the environment. Proc. 12th Symp. Eur. Grassl. Fed., pp. 303-306. Pleven, Bulgaria.

Van-Soest PJ, Robertson HB, Lewis BA. 1991. Method of dietary fiber and non-starch

polysaccharides determination in relation to animal material. J. Dairy. Sci. (74): 3583-

3591.

Wang Y, McAllister TA, Xu ZJ, Gruber MY, Skadhauge B, Jende-Strid B, Cheng KJ. 1999.

Effects of proanthocyanidins, dehulling and removal of pericarp on digestion of barley

grain by ruminal micro-organisms. J. Sci. Food Agric. 79: 929−938.

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45

Table 3.1. Date of sowing and harvest stage of maturity for three cereal fodders

Fodder type

Date of sowing

Date of harvest

Early bloom

(20% flowering)

Mid bloom

(50% flowering)

Full bloom

(100% flowering)

Maize 15 July 25 September 1st October 6th October

Sorghum 15 June 15th August 25 August 4th September

Oats 15 November 25 February 7th March 12th March

Table 3.2. Effect of maturity stages on chemical composition and nutritive value of silages

Maturity stages

Silages Parameters Early bloom Mid bloom Full bloom

Maize

DM% 18.82±0.02c 22.38±0.03b 25.80±0.05a

CP% 9.84±0.02a 8.60±0.01b 7.89±0.02c

NDF% 66.26±0.01b 66.83±0.01a 62.3±0.01c

ADF% 33.25±0.03b 34.76±0.02a 31.6±0.03c

ME(Mcal/kg) 2.92±0.003a 2.87±0.001b 2.85±0.02b

Sorghum

DM% 20.65±0.01c 26.56±0.03b 29.47±0.01a

CP% 7.45±0.31a 7.00±0.33ab 6.19±0.32b

NDF% 62.42±0.04b 56.50±0.08c 64.68±0.14a

ADF% 33.68±0.01b 29.38±0.01c 35.25±0.05a

ME (Mcal/kg) 2.83±0.009a 2.79±0.005b 2.78±0.009b

Oats

DM% 17.68±0.01c 23.61±0.03b 26.54±0.01a

CP% 8.86±0.02a 7.40±0.02b 6.82±0.01c

NDF% 52.70±0.36c 63.31±0.11b 65.33±0.03a

ADF% 33.42±0.03b 33.41±0.03b 34.26±0.02a

ME (Mcal/kg 2.80±.003a 2.79±0.005a 2.77±0.002b

Means within each row followed by different superscripts are significantly different (P<0.05).

Experiment 1

46

Table 3.3. Fermentation characteristics and IVDMD of silages at different stage of

maturity

Maturity stages

Silages parameters Early bloom Mid bloom Full bloom P-V

Maize

pH 4.29±0.005a 4.24±0.005b 3.94±0.008c <.0001

LA% 4.03±0.04c 4.46±0.08b 7.06±0.05a <.0001

IVDMD% 68.80±0.17 67.53±0.26 67.00±0.76 0.087

Sorghum

pH 3.95±0.01a 3.83±0.008b 3.62±0.017c <.0001

LA% 4.67±0.09c 5.76±0.09b 6.18±0.13a 0.0002

IVDMD% 65.30±0.90a 63.03±0.78a 60.26±0.49b 0.009

Oats

pH 4.04±0.008a 3.95±0.02b 3.71±0.008c <.0001

LA% 3.22±0.067c 3.74±0.19b 4.91±0.07a 0.0002

IVDMD% 60.60±0.49a 57.46±0.76b 55.33±0.41c 0.002

Means within each row followed by different superscripts are significantly different (P<0.05).

Experiment 1

47

Table 3.4. Effect of maturity stages on physical quality of silages

Maturity stages

Silages Parameters Early bloom Mid bloom Full bloom P-V

Maize

Color 3.66±0.08 3.50±0.05 3.70±0.05 0.172

Smell 3.76±0.14b 4.77±0.14a 5.27±0.40a 0.018

Structure 2.63±0.08c 3.43±0.12b 3.90±0.05a 0.000

Sensoric score 10.05 11.7 12.87

Flieg score 71.04 80.16 99.00

Quality class Good Very good Very good

Sorghum

Color 3.36±0.08 3.66±0.08 3.36±0.08 0.083

Smell 4.31±0.09b 4.40±0.09ab 4.91±0.21a 0.056

Structure 2.80±0.11c 3.37±0.17b 3.88±0.04a 0.002

Sensoric score 10.47 11.43 12.15

Flieg score 78.76 94.22 109.68

Quality class Good Very good Very good

Oats

Color 4.53±0.14 4.10±0.20 4.70±0.32 0.26

Smell 3.33±0.08b 3.40±0.05b 3.90±0.05a 0.002

Structure 2.80±0.15c 3.36±0.08b 3.83±0.08a 0.002

Sensoric score 10.66 10.86 12.43

Flieg score 88.3 104.92 119.14

Quality class Good Very good Very good

Means within each row followed by different superscripts are significantly different (P<0.05).

48

CHAPTER 4

EXPERIMENT-2

Comparison of silo types on chemical composition and physical quality of silage made from

maize, sorghum and oats fodders

ABSTRACT

The objective of current study was to investigate the effects of trench, bunker and plastic bag silos

on chemical and physical characteristics of silages made from three cereal fodders i.e. maize,

sorghum and oat in subtropical conditions. Each fodder was harvested at 30-35% dry matter (DM)

and ensiled in the said three silo types. The results revealed that trench silo had significantly

(P<0.05) highest sensory score (smell, color and structure) followed by bunker and bag silo for

each cereal silage. The sensory score for maize (12.99, 12.09 and 10.96), sorghum (11.17, 12.72,

and 10.28) and oat silages (12.55, 13.27 and 11.47) corresponding to trench, bunker and bag silos,

respectively. The lowest pH values were observed in trench followed by bunker and bag silos, in

maize (3.61, 3.65 and 3.81), sorghum (3.71, 3.81 and 3.89) and oat silage (3.82, 3.87 and 3.93),

respectively. However, the DM and crude protein (CP) were significantly (P<0.05) higher in trench

followed by bunker and bag silos. In-vitro dry matter digestibility (IVDMD) also varied

significantly (P<0.05) among silo types. The higher IVDMD % for maize (65.83, 64.53 and 63.00),

sorghum (62.23, 60.43 and 58.00) and oats (59.60, 58.6 and 57.16) silages were observed in trench

followed by bunker and bag silo, respectively. The current findings revealed that silage quality

was highest in trench silo for cereal fodders in subtropical conditions.

Experiment 2

49

INTRODUCTION

High quality silage is the result of several management practices. Johnson and Harrison

(2001) classified the management of silages into four categories: 1) harvesting 2) silo types 3)

filling and covering, and 4) feed out period. The stage of maturity at harvest has a marked impact

on fermentation qualities, digestibility and nutritive value of silages as reviewed in previous

chapter. The silo type also affects the physical and chemical properties of silages. Different types

of silos are in practice for silage making including bunker, pile, upright, pit or trench silo and

plastic bag systems.

The increased dry matter losses during ensiling period are often due to exposure to oxygen.

The pile and bunker silos have higher risk of oxygen exposure as compared to bag silos due to

increased surface area (Johnson and Harrison., 2001). The temperature during ensiling and feed

out period also impacts the silage quality. Johnson and Harrison (2001) observed that bag silo was

cooler during six month ensiling period and feed out phase compared to bunker silo. Also the cost

of silage production including harvesting and storage is an important factor to choose silo types.

Holmes (1998) conducted the cost analysis for different silo types in USA and found that bagging

system was the least cost as compared to pile, bunker or upright silo. However, such studies cannot

be generalized globally as the cost of raw material could vary area to area and may have different

the economics of silage production under similar silo types.

Most of the silage experiments were carried out in laboratory scale silos, and little data is

available mentioning the change in silage characteristics when moving from laboratory scale to

large scale silos. Laboratory scale silos are usually kept at room temperature, but large scale silos

are under different environmental conditions determined by the location and season (Kızılsimsek

et al. 2005). As silage production is getting popular in Pakistan and different silo types are in

practice however, the studies evaluating the effect of silo types on silage characteristics are limited.

Experiment 2

50

The objectives of the present study was to investigate the effects of silo types (bunker,

trench/pit, and bag) on the chemical composition, fermentation characteristics and physical quality

of maize, sorghum and oats silages.

MATERIALS AND METHODS

Fodder crops

The three fodder crops i.e. maize (Zee maize), sorghum (Sorghum bicolor) and oats (Avena

sativa) were used for silage making. The sorghum, maize, and oats were planted during the month

of June, July and November 2012, respectively on agriculture field of Dairy Animals Training

and Research Center, University of Veterinary and Animal Sciences, Ravi Campus Pattoki,

Pakistan (31°1'0" North, 73°50'60" East, 186 meters elevation).

Harvesting of fodder crops

All the crops under investigation in the field were harvested after full bloom with an

average dry matter of 30-35% at ensiling. For DM %, the respective fodder was randomly cut

during a clear day from four different parts of the field, chopped, mixed carefully, and duplicate

samples 250 g were dried in a hot oven at 60 ° C for 72 hours. The detail of planting and harvesting

has been presented in Table 4.1. The fodders were chopped by mechanical chopper (Fimax, V-

Belt Driven, MC 10X, Turkey) with a chop size of about 2 cm to makes it easy for compacting the

silage and removing air when loaded into silos.

Ensiling of fodders in bunker, trench and bag silos

The fodder crops were ensiled in three different silo types: 1) bunker; 2) trench/pit; 3)

plastic bags. The bunker, trench and plastic bag silos had the dimensions as 30×12×6, 30×12×6

feet and 36 × 24ʺ with the loading capacity of 40, 40 tones, and 40 kg of fodder, respectively. The

density of chopped fodder in silo (about 20 kg per cubic feet) was same for all types of silos.

Fodder was filled into the silos layer by layer compacted every layer by continuous treading to

Experiment 2

51

removes air and the silos were sealed immediately with an air-tight cover once it was filled to

inhibit the infiltration of air and precipitation in the silage for improving and speeding up

fermentation process. After 30 days of fermentation period, the three solos were opened and

samples were taken for physical quality, chemical composition fermentation characteristics and in

vitro dry matter digestibility (IVDMD).

Physical quality of silages

For physical analysis, the quality of silages was determined by color, smell, and structure

along with total flieg score described by Kilic (1986). For color evaluation, the scale 1- 4 was

used on the basis of change in green color from dark brown , dark green to pale yellow; for smell

, the scale 1-7 was used on the basis of repugnant putrid smell to acidic sweet pleasant smell; for

structure, the scale 1-4 was used on the basis of softness of leaves and stem as well as its ability to

remain intact after squeezing the silage tightly in hand and then opening from breaking into small

pieces to break into two or three pieces. The same person scored the silages for smell, color and

structure to avoid any bias. All the scores for color, smell and structure were added to make a

cumulative score as sensory score. Flieg score was calculated using a formula (flieg Score = 220

+ (2 x Dry Matter% - 15) - 40 x pH) reported by Kilics (1986). The flieg score with value 81-100,

61-80, 41-60, 21-40 and 0-20 represented the silage quality a very good, good, medium, low and

poor, respectively.

Chemical composition of silages

For chemical composition, approximately, 250g sample (in triplicate) was taken from each

silo type, dried in a hot-air oven (Memmert, Beschickung-Loading Model 100-800, Germany) at

60°C for 72 hours (for DM%), then ground through hammer mill (Wiley laboratory Mill, Standard

Model No. 2, Arthur H. Thomas Company, USA) making particle size of about 0.5 to1mm and

stored in pre labeled bottles for further laboratory analyses. Nitrogen (N) contents of samples were

determined by procedure AOAC. (1990) using Kjeldahl apparatus (ID 984.13), and then

Experiment 2

52

multiplying the N concentration by a factor 6.25 to calculate CP. The NDF and ADF contents were

determined according to Van Soest et al. (1991). The gross energy of the silage samples was

determined through the IKA C-2000 Bomb Calorimeter, while metabolizable energy (ME) was

calculated as 63% of the gross energy (Mandal et al, 2003).

Fermentation Characteristics

For fermentation characteristics the pH and lactic acid content was measured in silages.

Approximately 25g composite sample from different points of was taken from each silo type

immediately after opening. The sample silage was mixed with 100 ml of distilled water (Hart and

Horn, 1987). After hydration for 10 min using blender, the diluted material was then filtered

through cheese cloth and then pH was determined by using a digital pH meter. The liquid obtained

was further filtrated through Whatman 54 filter paper, centrifuged and kept at 20 ̊ C for lactic acid

determination by high pressure liquid chromatography (Muck and Dickerson, 1988).

In vitro dry matter digestibility of silages

The in vitro dry matter digestibility trials were conducted at University of Sydney, Camden.

The dried samples were taken from Pakistan to Camden by air cargo. For IVDMD study, rumen

liquor (inoculant) was collected from rumen of cannulated lactating Holstein cows managed on

pasture and cereal-based concentrate (9kg DM/cow/day), at Corstorphine farm, University of

Sydney. The collected rumen liquor was filtered through various layers of cheese cloth and mixed

with buffered minerals solution in 1:2 ratio and placed at 39 ̊ C under O2 free environment. Dry

matter digestibility (DMD) was determined in vitro by batch incubation of samples in rumen liquor

(Wang et al., 1999). All the dried samples from respective cereal silages were incubated in

duplicate using ANKOM filter bags (F57 filter bags; 128 pore size 25μm, 55 mm long and 50 mm

wide, New York, USA). The open side of the bag (having 0.5g ground sample) was sealed with

heat sealer impulse, and then put into a 50ml dark bottle. The bottle contained 25 ml of a 2: 1

buffer: rumen fluid saturated with gas N (O2-free) with 0.5 ml cysteine sulphide reducing agent.

Experiment 2

53

Bottles were fitted with rubber plugs placed in an incubator (Forma Scientific, model 39419-1,

Marietta, OH, USA). Incubator temperature was 39 C and bottles were placed at a rotary shaker

with 90 oscillations / min (Lab-Line Instruments Inc., Melrose Park, IL, USA). Eight bottles

containing only inoculum also included in each series as a blank control. After 48 h of incubation

the bags having digested sample were removed from the flasks, washed under running tap water

then dried in oven at 60 ̊ C for 48 hours. The IVDMD% was calculated from the difference of the

dry weight of sample and residues remained in the bag after 48 h of digestion divided by weight

of sample×100 (Wang et al., 1999).

Statistical Analysis

The collected data on different factors were analyzed through “complete randomized

design” (CRD) with one-way analysis of variance, using SAS 9.1.3 portable software. While,

Fisher LSD test was applied for means comparison (Steel et al., 1997).

RESULTS

Physical quality and fermentation characteristics of silages

The results indicated that trench silo had highest sensory score (smell color and structure)

followed by bunker and bag silo for each cereal silage (Table 4.2; P<0.05). The sensory score for

maize silage was 12.99, 12.09 and 10.96 in trench, bunker, and bag silo respectively. Similarly,

the sensory scores for sorghum and oats were 11.17, 12.72, 10.28 and 12.55, 13.27 11.47

corresponding to trench, bunker and bag silos, respectively. Also, the highest flieg score was

observed in trench followed by bunker and bag silo irrespective of the cereal fodder. Flieg score

for maize (118.08, 121.12 and 109.08), sorghum (110.22, 116.92 and 106) and oats silages

(106.04, 108.66 and 102.66) were presented in Table 4.2.

The results showed that silo type had significant effect on lactic acid concentration and pH

values (Table 4.3; P<0.05). The respective lactic acid contents for maize, sorghum and oats were

Experiment 2

54

(8.65, 9.19, 8.38), (6.40, 6.52 6.37) and (5.80, 5.85, 5.75) corresponding to bunker, trench and bag

silos, respectively. Similarly, the lowest pH values was observed in trench followed by bunker and

bag silos, in maize (3.61, 3.65 and 3.81), sorghum (3.71, 3.81 and 3.89) and oat silage (3.82, 3.87

and 3.93) respectively.

Chemical composition and In-vitro dry matter digestibility of silages

The DM and CP were significantly (P<0.05) higher in trench followed by bunker and bag

silos made from maize, sorghum and oats fodders respectively (Table 4.4). However, silo type did

not have any effect on NDF and ADF concentration in all cereals silages (Table 4.4).

Silo types significantly (P<0.05) affected the IVDMD in ensiled cereals (maize, sorghum

and oats). The higher IVDMD digestibility was observed in trench followed by bunker and bag

silo for maize (65.83, 64.53 and 63.00), sorghum (62.23, 60.43 and 58.00) and oats (59.60, 58.6

and 57.16) silages, respectively (Table 4.3).

DISCUSSION

Physical quality and fermentation characteristics of silages

The higher sensory and flieg scores, and low pH for trench silos compared to bunk and bag

silos were in line with Mtengeti et al. (2014) who reported that overall quality (flieg score and

sensory scores) of elephant grass silage ensiled in trench silos was slightly better than concrete

bunker silos. They also reported low pH for trench silos. This was probably due to underground

cool environment of the trench silos, whereas the bunk silos were built above the ground and were

more exposed to direct changes in ambient temperatures thereby increasing chances of the walls

of the silos to absorb the excess heat and cold that might have affected the normal microbial

fermentation. Also, the trench silo might have facilitated better packing and compaction of forage

material inside the silo. Similarly, the current findings were in agreement to Kızılsimsek et al.

(2005) who found lower pH values in big scale silo as compared to laboratory scale silo of winter

and spring leguminous and cereals silages and suggested that silages in big scale were better

Experiment 2

55

fermented than in laboratory. The lower pH is usually an indicative of increased lactic acid

concentration thereby implying better fermentation of silages during ensiling period.

Chemical composition and In-vitro dry matter digestibility of silages

The results of present study indicating the higher DM and increased in-vitro DM

digestibility in trench silo were similar to the previous studies. Mtengeti et al. (2014) reported that

DM and as CP contents, were higher in elephant grass silage ensiled in trench silos compared to

concrete bunker silos. Pizarro and Vera (1980) also studied the effect of silo types in maize fodder

and found that DM losses were lowest (9%) in trench silo compared to bunker (25%) and clamp

silos (35%). The abnormal bacterial fermentation due to change in environmental temperature

could be a reason for higher DM in trench silos as they are built in ground and have more stable

ambient temperature. Contrary to current findings, Johnson and Harrison (2001) reported that DM

losses were higher in bunker silos than bag silos. They did not compare the trench silo with other

types. Although they attributed the increased loss of DM in bunker silo to more exposed surface

area to oxygen in bunker silo as compared to bag silo however, the better results in their study for

bag silos could also be due to the size of bag silos as large scale silos tended to have better

fermentation as describe earlier.

Conclusion

Considering the current findings, it was concluded that trench silos were better in making

silages from cereal fodders as fodders ensiled in trench silos have better physical quality, chemical

composition and fermentation characteristics of silages. It seemed that trench silos more resistant

to ambient temperature there by improving the silage quality in sub-tropical area like Pakistan.

Experiment 2

56

LITERATURE CITED

AOAC (Association of Official Analytical Chemists). 1990. Official Methods of Analysis. 15th

Ed., Association of Official Analytical Chemists, Arlington, Virginia, USA.

Holmes BJ. 1998. Choosing forage storage facilities. Proceedings in Dairy Feeding Systems,

Management, Components, and Nutrients Conference. NRAES-116. Ithaca, NY.

Hart SP, Horn FP. 1987. Ensiling characteristics and digestibility of combinations of turnips and

wheat straw. J. Anim. Sci., 14: 1790-1800

KızılsimsekM, Erol A,Calıslar S. 2005. Effects of raw material and silo size on silage quality.

Livestock research for rural development, 17 (3).

Kilic A. 1986. Silo feed (Instruction, Education and Application Proposals). Bilgehan Press,

Izmir, pp: 327.

Mtengeti EJ, MaedaFH, UrioNA. 2014. Effects of chopping, additive and silo type on the quality

of elephant grass (Pennisetum Purpureum) silage. Livestock Research for Rural

Development 26 (4).

Mandal AB, Paual SS, Pathak NN. 2003. Nutrient requirements and feeding of buffaloes and cattle.

Published by Int. Book Distributing Co. Charbagh, Lucknow, India. P-23.

Muck RE, Dickerson JT. 1988. Storage temperature effects on proteolysis in alfalfa silage. Trans.

ASAE. 31:1005-1009.

Johnson LM, Harrison JH. 2001. Scientific aspects of silage making. Proceedings, 31st California

Alfalfa & Forage Symposium, (12-13, December), Modesto, CA, UC Cooperative

Extension, University of California, Davis.

Pizaro EA, Vera R R. 1980. Efficiency of fodder conservation systems. Maize silage. In: Fodder

Conservation in the 80’s (Edited by Thomas C). Occasional Symposium No.11. British

Grassland Society. Pp 436-441.

Experiment 2

57

Steel GD, Torrie JH, Dickey DA. 1997. Principles and procedures of statistics, 3rd ed., Mc Graw-

Hill, New York.

Van-Soest PJ, Robertson HB, Lewis BA. 1991. Method of dietary fiber and non-starch

polysaccharides determination in relation to animal material. J. Dairy. Sci. (74): 3583-

3591.

Wang Y, McAllister TA, Xu ZJ, Gruber MY, Skadhauge B, Jende-Strid B, Cheng KJ. 1999.

Effects of proanthocyanidins, dehulling and removal of pericarp on digestion of

barley grain by ruminal micro-organisms. J. Sci. Food Agric. 79: 929−938.

Experiment 2

58

Table 4.1. Date of sowing and harvest for three cereal fodders

Fodder type Date of sowing Date of harvest

Maize 15 July 21 October

Sorghum 15 June 19 September

Oats 15 November 28 March

Table 4.2. Effects of silo types on physical characteristics of cereal silage

Silo type

Silages Parameters Bunker Trench Bag P-value

Maize

Color 3.54±0.05b 3.80±0.02a 3.27±0.04c 0.0004

Smell 5.64±0.03a 5.71±0.02a 4.93±0.05b <.0001

Structure 2.91±0.12b 3.48±0.08a 2.76±0.16b 0.0187

Sensory score 12.09 12.99 10.96

Flieg score 118.08 121.12 109.08

Sorghum

Color 3.40±0.008b 3.52±0.01a 3.32±0.04b 0.0046

Smell 5.20±0.25b 6.13±0.20a 4.50±0.17b 0.0046

Structure 2.57±0.03b 3.07±0.12a 2.46±0.20b 0.0467

Sensory score 11.17 12.72 10.28

Flieg score 110.22 116.92 106

Oats

Color 3.79±0.02b 3.92±0.01a 3.72±0.01b 0.0014

Smell 5.33±0.27a 5.73±0.23a 4.53±0.14b 0.0235

Structure 3.43±0.04ab 3.62±0.10a 3.22±0.05b 0.0218

Sensory score 12.55 13.27 11.47

Flieg score 106.04 108.66 102.66

Means within each column followed by different superscripts are significantly different (P<0.05).

Experiment 2

59

Table 4.3. Effects of silo types on fermentation characteristics and IVDMD of silages

Silo type

Silage Parameters Bunker Trench Bag p-value

Maize

pH 3.65±0.01b 3.61±0.01b 3.81±0.02a 0.0004

Lactic acid % 8.65±0.06b 9.19±0.06a 8.38±0.08c 0.0005

IVDMD% 64.53±0.14b 65.83±0.33a 63.00±0.36c 0.0016

Sorghum

pH 3.81±0.005b 3.71±0.01c 3.89±0.01a 0.0001

LA 6.40±0.16a 6.52±0.21a 6.37±0.13a 0.8020

IVDMD% 60.43±0.44b 62.23±0.21a 58.00±0.20c 0.0002

Oats

pH 3.87±0.005b 3.82±0.01c 3.93±0.008a 0.0012

LA 5.80±0.17a 5.85±0.18a 5.75±0.19a 0.9251

IVDMD% 58.6±0.27a 59.60±0.36a 57.16±0.37b 0.0065

Means within each row followed by different superscripts are significantly different (p<0.05).

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Table 4.4. Effects of silo type on chemical composition of cereals silages

Silo type

Silage Parameters Bunker Trench Bag p-value

Maize

DM% 29.66±0.08b 30.26±0.04a 28.24±0.04c <.0001

CP% 6.19±0.02b 6.58±0.08a 6.15±0.03b 0.0021

NDF% 62.97±0.24a 63.70±0.66a 62.97±1.06a 0.7353

ADF% 32.47±0.18a 32.64±0.22a 33.23±0.63a 0.4312

ME(Mcal/kg) 2.87±0.006b 2.93±0.02a 2.85±0.005b 0.0238

Sorghum

DM% 28.81±0.10b 30.16±0.04a 28.30±0.08c <.0001

CP% 5.58±0.06b 5.85±0.05a 5.31±0.02c 0.0008

NDF% 61.47±1.96a 59.84±2.01a 61.10±2.42a 0.8572

ADF% 29.77±2.75a 30.02±2.49a 32.34±2.19a 0.7363

ME(Mcal/kg) 2.84±0.001a 2.83±0.002a 2.56±0.23a 0.3321

Oats

DM 27.92±0.03b 28.23±0.05a 27.43±0.05c <.0001

CP 5.70±0.04b 5.92±0.04a 5.60±0.02b 0.0039

NDF 63.11±0.43a 64.15±0.15a 61.33±2.23a 0.3716

ADF 34.22±0.52a 34.21±0.96a 33.93±0.73a 0.9527

ME(Mcal/kg) 2.78±0.01b 2.84±0.001a 2.82±0.002a 0.0099

Means within each row followed by different superscripts are significantly different (p<0.05).

61

CHAPTER 5

EXPERIMENT-3

Study of different levels of commercial additive “Sil-all” on quality of cereals silages

ABSTRACT

The present study examined the effect of silage biological additive (Sil-all 4 × 4) on nutritive value,

fermentation and physical quality of silages made from maize, sorghum and oat fodders. All the

cereal fodders were harvested having 30-35% DM contents and ensiled in laboratory silos with

additive at the rate of 0.0(control), 8, 10 and 12g/ton fresh forage material for 35days of ensilation

period. The statistical analysis of variance exposed that with increasing level of additive

significantly (P<0.05) increased DM% in Maize (29.01±0.006, 30.53±0.02, 31.92±0.005 and

31.69±0.008), Sorghum (28.28±0.01, 29.52±0.06, 30.17±0.03 and 30.76±0.02) and Oat

(29.65±0.06, 30.54±0.04, 30.15±0.02, and 30.72±0.03) respectively. Whereas, a similar pattern

was also observed for CP content in all. While, in contrast NDF and ADF concentration decreased

significantly (P<0.05) with increasing additive levels. The decreased of NDF in maize

(64.32±0.01, 57.88±0.01, 56.21±0.02 and 49.95±0.02), Sorghum (64.68±0.73, 63.17±0.03,

62.86±0.03 and 58.54±0.32) and Oat (65.26±0.08, 63.30±0.10, 60.87±0.32 and 59.13±0.19)

silages, a similar trend was also revealed for ADF content in all. However, the pH recorded on day

15, 18, and 22 as well as after 35 days of ensilation was also significant (P<0.05) among the

treatments in all silages. Lactic acid concentration and in-vitro dry matter digestibility (IVDMD)

were significantly (P<0.05) higher in inoculated as compare to control silage. Numerically the

highest flieg score regarding to silage quality were recorded in T2 followed by T3, T1 and T0

(control) silages. The results of the current study indicate that cereal fodders ensiled with 10g/ton

of sil-all additive could be economical in terms of nutrients recovery.

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62

INTRODUCTION

Considering the actual weather conditions, silage is the best method for preserving fresh

forage with minimal losses. The quality and nutritive value are influenced by many biological and

technological factors. When appropriate techniques are used, silage will have high nutritional

value and quality. The silage quality is often poor or unsatisfactory when the fermentation

conditions are not fully met (Lattemae et al., 2006). Factors that influence the degree of

fermentation include wilting green forage, cut length, ensiling type of technology, and the amount

of an additive used (Haigh., 1988). Silage additives include feedstuffs, urea, inoculants and acids

(Weiss and Underwood., 2006). The major goal in silage making is to preserve silage material

with minimum nutrient loss. In order to achieve this goal, growth of acid producing bacteria should

be stimulated. Especially, lactic acid producing bacteria is generally used to accomplish this target.

Wheat is usually used to deliver readily available carbohydrates, needed for fermentation process

during ensiling and commercial bacterial inoculant is used to create a desirable microbial

population to convert energy into organic acids ultimately reducing pH in ensiled forage.

Cereal fodders especially maize is an ideal crop for silage making due to comparatively

high dry matter (DM) content, acceptable crude protein (buffering capacity) and adequate water-

soluble carbohydrates (energy) for lactic acid production (McDonald et al., 1991). To inhibit the

growth of enterobacteria and clostridia bacteria, a rapid drop in pH is needed, for achieving high

quality well fermented palatable silage (McDonald et al., 1991). This happens when native

homofermentative acids producing bacteria utilize water-soluble carbohydrates and produce

organic acids (lactic and acetic acids). However, heterofermentative lactic acid bacteria are

dominant on a cereals crop prior to ensiling, fermentation will be less efficient and the end products

of fermentation will be lactic acid, acetic acid, ethanol and carbon dioxide (McDonald et al., 1991).

The population of lactic acid bacteria present on maize plants prior to ensiling is too low which is

not sufficient for conversion of energy into organic acids. Speckman et al. (1981) surveyed

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63

numbers of lactobacilli on maize crops in the USA and showed that 69% of samples had counts

below 1000 colony forming units per gram of fresh material. However, Meeske & Basson (1998)

found that the number of lactic acid bacteria on fresh chopped maize plants prior to ensiling was

as high as 109colony forming units per gram of fresh material.

Ensiling phenomenon is based on natural anaerobic fermentation of fodder in presence of

lactic acid producing bacteria, which converts readily fermentable carbohydrates into organic acids

(Koc et al., 2008). During this process water soluble carbohydrates are respired and intrinsic plant

proteases can convert the protein into ammonia (Muck., 1988). Early achieved anaerobic

conditions and rapid decline in pH can minimize the nutrient losses by reducing respiration and

prolonged fermentation (Charmley., 2001). So, a rapid decline in silage pH with the addition of

inoculants can improve the fermentation characteristics, nutritive value and utilization of the

silage.

Therefore, the current study was planned to determine the effect of adding three inclusion

level of sil-all (a commercial biological silage inoculant) on cereal fodders maize, sorghum and

oat at the time of ensiling on fermentation characteristics, chemical composition and physical

quality of the silages.

MATERIALS AND METHODS

Fodder crops

The three fodder crops i.e. maize (Zee maize), sorghum (Sorghum bicolor) and oats (Avena

sativa) were used for silage making. The detail of planting and harvesting has been presented in

Table 5.1. The maize, sorghum, and oats were planted during the month of June, July and

November 2012, respectively on agriculture field of Dairy Animals Training and Research Center,

University of Veterinary and Animal Sciences, Ravi Campus Pattoki, Pakistan (31°1'0" North,

73°50'60" East, 186 meters elevation). The fodder crops were harvested with sickle after full bloom

Experiment 3

64

with an average dry matter of 30-35% and then chopped by mechanical chopper (Fimax, V-Belt

Driven, MC 10X, Turkey) with a chop size of about 2 cm.

Inoculants and treatment groups

A commercial biological additive (Sil-All 4 × 4, Alltech) containing homo and hetero-

fermentative lactic acid producing bacteria (L. plantarum, Enterococcus faecium, Pediococcus

acidilacti, and Lactobacillus salivarius) and 4 enzymes (α-amylase, cellulase, hemicellulose and

pentosanase) was used for inoculation. Each fodder crop was divided into 4 groups; 1) T0 the

silage was made without the addition of inoculants; 2) T1 inoculants was added at 8g/ton on fresh

forage material; 3) T2 10g/ton on fresh forage; 4) T3 12g/ton on fresh forage material.

Application of inoculants on fodders and ensiling

The chopped material of maize, sorghum, and oats forages was weighed and spread out on

a 5×5 meter plastic sheet for each treatment group separately. For inoculation purpose a fresh

inoculant culture of “Sil-All 4 × 4” was first dissolved separately in 200 ml of distilled water

according to mentioned dose (8, 10 and 12g/ton) and then sprayed the whole suspension evenly

onto each mass of respective forage and one treatment was made as control sprayed with 200ml of

distilled water (no-inoculant) then mixed manually by rolling the forage on the plastic sheet. The

treated forages of each respective treatments were ensiled in a pre-labeled polyethylene bags silos

with loading capacities (35-40 kg), having dimensions 80×40 cm. All the bags were sealed

immediately and stored under shed at room temperature for fermentation.

During fermentation period a random sample was taken from each treatment on day 15, 21

and 28, for determination of pH. After 30 days of ensiling period all bags of respective silages

were opened and a composite sample was taken for physical quality, chemical composition

fermentation characteristics and in-vitro dry matter digestibility (IVDMD).

Experiment 3

65

Physical quality of silages

For physical analysis, the quality of silages was determined by total flieg score described

by Kilic (1986). Flieg score was calculated using a formula (flieg Score = 220 + (2 x Dry Matter%

- 15) - 40 x pH) reported by Kilics (1986). The flieg score with value 81-100, 61-80, 41-60, 21-40

and 0-20 represented the silage quality a very good, good, medium, low and poor, respectively.

Chemical composition of silages

For chemical composition, approximately, 250g sample (in triplicate) was taken from each

silo type, dried in a hot-air oven (Memmert, Beschickung-Loading Model 100-800, Germany) at

60°C for 72 hours (for DM%), then ground through hammer mill (Wiley laboratory Mill, Standard

Model No. 2, Arthur H. Thomas Company, USA) making particle size of about 0.5 to1mm and

stored in pre labeled bottles for further laboratory analyses. Nitrogen (N) contents of samples were

determined by procedure AOAC. (1990) using Kjeldahl apparatus (ID 984.13), and then

multiplying the N concentration by a factor 6.25 to calculate CP. The NDF and ADF contents were

determined according to Van Soest et al. (1991). The gross energy of the silage samples was

determined through the IKA C-2000 Bomb Calorimeter, while metabolizable energy (ME) was

calculated as 63% of the gross energy (Mandal et al., 2003).

Fermentation Characteristics

For fermentation characteristics the pH and lactic acid content was measured in silages.

Approximately 25g composite sample from different points of was taken from each silo type

immediately after opening. The sample silage was mixed with 100 ml of distilled water (Hart and

Horn., 1987). After hydration for 10 min using blender, the diluted material was then filtered

through cheese cloth and then pH was determined by using a digital pH meter. The liquid obtained

was further filtrated through Whatman 54 filter paper, centrifuged and kept at 20 ̊ C for lactic acid

determination by high pressure liquid chromatography (Muck and Dickerson., 1988).

Experiment 3

66

In vitro dry matter digestibility of silages

The in-vitro dry matter digestibility trials were conducted at University of Sydney,

Camden. The dried samples were taken from Pakistan to Camden by air cargo. For IVDMD study,

rumen liquor (inoculant) was collected from rumen of cannulated lactating Holstein cows managed

on pasture and cereal-based concentrate (9kg DM/cow/day), at Corstorphine farm, University of

Sydney. The collected rumen liquor was filtered through various layers of cheese cloth and mixed

with buffered minerals solution in 1:2 ratio and placed at 39 ̊ C under O2 free environment. Dry

matter digestibility (DMD) was determined in vitro by batch incubation of samples in rumen liquor

(Wang et al., 1999). All the dried samples from respective cereal silages were incubated in

duplicate using ANKOM filter bags (F57 filter bags; 128 pore size 25μm, 55 mm long and 50 mm

wide, New York, USA). The open side of the bag (having 0.5g ground sample) was sealed with

heat sealer impulse, and then put into a 50ml dark bottle. The bottle contained 25 ml of a 2: 1

buffer: rumen fluid saturated with gas N (O2-free) with 0.5 ml cysteine sulphide reducing agent.

Bottles were fitted with rubber plugs placed in an incubator (Forma Scientific, model 39419-1,

Marietta, OH, USA). Incubator temperature was 39 C and bottles were placed at a rotary shaker

with 90 oscillations / min (Lab-Line Instruments Inc., Melrose Park, IL, USA). Eight bottles

containing only inoculum also included in each series as a blank control. After 48 h of incubation

the bags having digested sample were removed from the flasks, washed under running tap water

then dried in oven at 60 ̊ C for 48 hours. The IVDMD% was calculated from the difference of the

dry weight of sample and residues remained in the bag after 48 h of digestion divided by weight

of sample×100 (Wang et al., 1999).

Experiment 3

67

Statistical Analysis

The collected data on different factors were analyzed under completely randomized design

with one-way analysis of variance, using SAS 9.1.3 portable software. The comparison of means

was done by DMR test (Steel et al., 1997).

RESULTS

Physical quality and fermentation characteristics of silages

Flieg score for maize (107.82, 114.06, 118.84 and 115.58), sorghum (93.16, 113.16, 112.94

and 112.92) and oats silages (111.9, 117.27, 114.9 and 116.44) were presented in Table 5.2.

Numerically the highest flieg score with respect to silage physical quality were recorded in T2

followed by T3, T1 and T0 (control) in Maize and sorghum silages. While in oats the highest flieg

score was recorded in T1 followed by T3, T1 and T0. The addition of biological additives at ensiling

enhanced the fermentation characteristics including pH and lactic acid of cereal silages. The pH

values significantly (P<0.05) decreased and lactic acid concentration increased in inoculated

compared to control silages. A significantly (P<0.05) lower pH was detected in inoculated one on

days 15, 21 and 28 during fermentation, as well as after 30 days of fermentation period as compare

to control silages (Table 5.3). However, non-significant (P>0.05) difference was found between

T2 and T3, while T0 was significantly (P<0.05) different with T1, T2 and T3 in maize, sorghum and

oats silages after 30 days of incubation. Lactic acid concentration was also significantly (P<0.05)

different between control and inoculated cereals silages, but non-significant (P>0.05) results was

observed between T2 and T3 but significant with T1 in all observation (Table 5.2).

Chemical composition and In-vitro dry matter digestibility of silages

The chemical composition of inoculated and un-inoculated maize, sorghum and oat silages

has been shown in Table 5.4. Inoculated silage had significantly (P<0.05) higher DM content then

control (T0) for all cereal silages. Also, DM content significantly (P<0.05) differed with the

Experiment 3

68

varying inoculants. Higher DM content was observed in T2 followed by T3 and T1 inoculated

treatments in all cases. A similar pattern for CP content was observed between control and

inoculated silage in all cases. While, in contrast to DM and CP content, a significantly (P<0.05)

lowest concentration of NDF and ADF were observed in inoculated as compare to control, and

also decreased significantly (P<0.05) with increasing level of inoculants at ensiling of the forages.

IVDMD was significantly higher (P<0.05) in inoculated treatments compared to control,

nonetheless the difference was non-significant between T1, T2 and T3 in maize silage. Similarly,

in sorghum a significant difference were observed between control and inoculated, but non-

significant difference between T2 and T3 while both are significant with T1 treatment. In oats

increasing inoculants level also increased IVDMD. The highest IVDMD was recorded in T3

followed by T2, T1 and T0 respectively (Table 5.2).

DISCUSSION

Physical quality and fermentation characteristics of silages

The findings of lower pH and increased lactic acid concentration in inoculated silages in

present study were in agreement to Nkosi et al. (2012) who studied the application of bacterial

inoculant and cellulase enzyme on fermentation quality of silage made from sorghum forage in

laboratory jars. They concluded that inoculation reduced pH, and increased lactic acid content in

inoculated silage compared with control silage. Similarly, Sucu and Filya (2006) reported that

higher lactic acid concentration and lower pH value was recorded in inoculated corn silage.

Likewise, Aragon et al. (2012) reported that inoculation of whole crop maize fodder at ensiling

with commercial additive (blend of homo- and hetero-fermentative lactic acid bacteria, BSM)

increased the fermentation rate with a significantly deeper pH and increased concentration of lactic

acid compared to un-treated. The bacterial inoculants stimulate lactic acid fermentation, increasing

speed of pH decrease and improving silage preservation.

Experiment 3

69

Chemical composition and In-vitro dry matter digestibility of silages

The results of our study were in agreement with the findings of Aragon et al. (2012) who

found that DM recovery and digestible protein was significantly (P<0.05) higher in maize silage

treated with commercial inoculant having bacteria “Enterococcus faecium, Lactobacillus

plantarum, and Lactobacillus brevis” in comparison with control silage (without

additives).Similarly, Iqbal et al. (2005) measured the effects of multiple probiotic (organic green

culture) and enzose (corn dextrose) on chemical composition of mott grass silage and reported that

DM and CP losses were decreased with increasing levels of multiple probiotic and enzose

concentration. The sharp decline in pH of inoculated silages was the major reason in reducing the

protein degradation during fermentation process (Xing et al. 2009) and thereby increasing DM in

inoculated silages. Contrary to current findings, Meeske et al. (2002) reported that CP

concentration was higher in control silage compared to the inoculated silage, and suggested that

protein breakdown or N loss was more in laboratory treated maize silage. This contradiction could

be due to the variation ensiling temperature as it could significantly affect fermentation process

and thereby CP and DM of silages.

In agreement to our findings Ozduven et al. (2010) investigated the application of enzymes

or lactic acid or mixture of both additives and reported the decrease neutral in NDF content and

increased in-vitro dry matter digestibility of triticale silages. However, Ozduven et al. (2010) also

reported that application of the above additive treatments did not affect ADF concentration in

triticale silage, contrary to our ADF results. The inoculants or application of enzymes (cellulases

and hemicellulases) degraded the cell wall content of the ensiled crops and subsequently improved

the organic matter and fiber digestibility (Selmer-Olsen et al., 1993).

Conclusion

The results of the current study indicate that cereal fodders ensiled with 10g/ton of sil-all

additive could be economical in terms of nutrients recovery.

Experiment 3

70

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and an enzyme on the fermentation quality and aerobic stability of ensiled whole-crop

sweet sorghum. S. Afr. J. Anim Sci. 42 (3):233-240.

Ozduven ML, Onal ZK, Koc F. 2010. The Effects of Bacterial Inoculants and/or Enzymes on the

Fermentation, Aerobic Stability and in vitro Dry and Organic Matter Digestibility

Characteristics of Triticale Silages. Kafkas Univ Vet Fak Derg.16(5):751-756.

Sucu E, Filya I. 2006. Effects of homo-fermentative lactic acid bacterial inoculants on the

fermentation and aerobic stability characteristics of low dry matter corn silages. Turk J Vet

Anim Sci, 30, 83-88.

Steel GD, Torrie JH, Dickey DA. 1997. Principles and procedures of statistics, 3rd ed., Mc Graw-

Hill, New York.

Experiment 3

72

Selmer-Olsen I, Henderson AR, Robertson S, McGinn R. 1993. Cell wall degrading enzymes for

silage. 1. The fermentation of enzyme-treated ryegrass in laboratory silos. Grass Forage

Sci, 48: 45-54.

Speckman CA, Phillips RM, Linnertz DP, Berger JCA, Carver LA, Parker RB. 1981. A survey for

indigenous Lactobacillus species on standing field corn at ensiling maturity. J. Anim. Sci.

53, Suppl., 1, 99.

Van-Soest PJ, Robertson HB, Lewis BA. 1991. Method of dietary fiber and non-starch

polysaccharides determination in relation to animal material. J. Dairy. Sci. (74): 3583-

3591.

Weiss B, Underwood J. 2006. Silage Additives. Ohio State University Extension department of

Horticulture and Crop Science 2021 Coffey Road, Colombus, Ohio 43210-1044, AGF-

018-92.

Wang Y, McAllister TA, Xu ZJ, Gruber MY, Skadhauge B, Jende-Strid B, Cheng KJ. 1999.

Effects of proanthocyanidins, dehulling and removal of pericarp on digestion of barley

grain by ruminal micro-organisms. J. Sci. Food Agric. 79: 929−938.

Xing L, Chen J, Han LJ. 2009. The effect of an inoculant and enzymes on fermentation and

nutritive value of sorghum straw silages. Bioresour Technol, 100, 488-491.

Experiment 3

73

Table 5.1. Date of sowing and harvest for three cereals fodders

Fodder type Date of sowing Date of harvest

Maize 15 July 21 October

Sorghum 15 Jun 19 September

Oats 15 November 28 March

Table 5.2. Effects of additive on fliege score, IVDMD and LA of silages

Inoculant level

Silages Parameters 0.0(control)

T

8g/ton

T1

10g/ton

T2

12g/ton

T3

Maize

Lactic acid% 7.32±0.25c 9.81±0.02b 10.27±0.02a 10.53±0.03a

IVDMD% 62.95±0.42b 64.50±0.17a 64.37±0.08a 64.87±0.22a

Flieg score 107.82 114.06 118.84 115.58

Sorghum

Lactic acid% 5.81±0.01d 8.33±0.05c 8.56±0.02b 8.87±0.04a

IVDMD% 56.27±0.74c 58.34±0.09b 60.56±0.19a 60.65±0.05a

Flieg score 93.16 113.16 112.94 112.92

Oats

Lactic acid% 4.90±0.02c 6.37±0.09b 6.97±0.39ba 7.70±0.24a

IVDMD% 54.44±0.66d 56.83±0.17c 58.34±0.09b 60.56±0.19a

Flieg score 111.9 117.27 114.9 116.44

Means within each row followed by different superscripts are significantly different (P<0.05).

Experiment 3

74

Table 5.3. Effects of additive on pH during fermentation kinetics of silage

Inoculants level

Silages

Days of ensiling

0.0(control)

(T)

8g/ton

(T1)

10g/ton

(T2)

12g/ton

(T2)

pH

Maize

15 4.17±0.02a 3.86±0.03c 4.02±0.04b 4.01±0.01b

21 3.94±0.008a 3.81±0.01b 3.86±0.02b 3.84±0.01b

28 3.84±0.01a 3.81±0.008ba 3.76±0.01bc 3.75±0.02d

After 30 days 3.88±0.02a 3.77±0.008c 3.82±0.008b 3.82±0.01b

Sorghum

15 4.16±0.02a 4.04±0.01b 4.03±0.01b 4.05±0.03b

21 4.04±0.03a 3.82±0.01bc 3.77±0.02c 3.88±0.01b

28 3.88±0.01a 3.80±0.02bc 3.75±0.02c 3.82±0.01ab

After 30 days 4.21±0.01a 3.76±0.01c 3.81±0.005b 3.84±0.01b

Oats

15 3.93±0.008a 3.75±0.01c 3.81±0.005b 3.84±0.01b

21 3.70±0.01a 3.53±0.01c 3.61±0.01b 3.63±0.02b

28 3.66±0.008a 3.51±0.008c 3.56±0.006b 3.54±0.01bc

After 30 days 3.81±0.005a 3.72±0.01c 3.76±0.008b 3.75±0.01b

Means within each row followed by different superscripts are significantly different (P<0.05).

Experiment 3

75

Table 5.4. Effects of inclusion level of additive on chemical composition of silages

ADDITIVE LEVEL

Silages Parameters 0.0(control)

T0

8g/ton

T1

10g/ton

T2

12g/ton

T3

Maize

DM% 29.01±0.006c 30.53±0.02b 31.92±0.005a 31.69±0.008a

CP% 6.05±0.006c 6.50±0.11a 6.20±0.05b 6.27±0.01bb

NDF% 64.32±0.01a 57.88±0.01b 56.21±0.02c 49.95±0.02d

ADF% 24.78±0.08a 23.09±0.45b 22.40±0.50b 23.46±0.08b

ME(Mcal/kg) 2.85±0.01a 2.85±0.01a 2.85±0.01a 2.84±0.01a

Sorghum

DM% 28.28±0.01d 29.52±0.06c 30.17±0.03b 30.76±0.02a

CP% 5.12±0.01c 5.65±0.02ab 5.76±0.02a 5.58±0.09b

NDF% 64.68±0.73a 63.17±0.03b 62.86±0.03b 58.54±0.32c

ADF% 33.08±0.68a 33.03±0.26a 32.52±0.65a 32.31±0.90a

ME(Mcal/kg) 2.80±0.002b 2.80±0.001ab 2.8±0.001a 2.80±0.002a

Oats

DM% 29.65±0.06d 30.54±0.04b 30.15±0.02c 30.72±0.03a

CP% 5.61±0.08b 6.33±0.06a 6.40±0.02a 6.23±0.11a

NDF% 65.26±0.08a 63.30±0.10b 60.87±0.32c 59.13±0.19d

ADF% 34.59±1.02a 35.40±1.52a 35.06±0.92a 35.40±0.90a

ME(Mcal/kg) 2.82±0.004a 2.82±0.007a 2.83±0.01a 2.83±0.005a

Means within each row followed by different superscripts are significantly different (P<0.05).

76

CHAPTER 6

EXPERIMENT-4

Effect of feeding maize, sorghum and oat silage on growth performance of Nili-Ravi buffalo

calves during summer in the sub-tropical region of Pakistan

ABSTRACT

Twenty four young male calves of Nili-Ravi buffalo with an average age of 7.0±1 months (mean

± SD) and weight 110±08kg were raised during summer of 2013 (14 May to 12 August), at Buffalo

Research Institute, Pattoki, District Kasur, Pakistan. All the calves were divided into four groups

according to completely randomized design, with six calves in each group. The groups and

treatments are as, A = Maize fodder, B= Maize silage C = Sorghum silage and D = Oats silage.

The results showed that dry mater intake (DMI) was significantly (P< 0.05) higher in calves of

group A, similar between B and C but lower in group D, stand as 3.38±0.04, 2.44±0.03,

2.43±0.02and 2.24±0.02kg/day respectively. While, crude protein (CP) intake was also higher in

group A, followed by B, C and D. Similarly, NDF and ADF intake were also significantly (P<0.05)

higher in group A, but nonsignificant between groups B, C and D. Numerically weight gain was

different however surprisingly non-significant (P > 0.05) among the groups. While dry matter

digestibility (DMD) revealed non-significant (P > 0.05) result between the groups. Numerically

low digestibility of control diet might be due to poor quality summer fodder as well as warm

environmental condition. However same pattern existed for CP and NDF digestibility among the

groups. Higher feed efficiency was observed in calves fed MS diet (0.167) to convert into gain

followed by SS (0.160), OS (0.157) and MF (0.110). The calves consumed more feed on MF diet

to gain one kg of body weight, while on MS followed by SS and OS diet performed best and were

highly efficient to gain in weight. It is concluded from the results of current study that cereal silages

could replace the summer grown maize fodder in the diet of growing buffalo calves without any

negative effect on growth and digestibility.

Experiment 4

77

INTRODUCTION

In Pakistan, majority of buffalo is raised on low quality forages and crop-waste that contain

high levels of lignocelluloses, low proteins and fermentable carbohydrates. Although feeding

fresh-green fodder (legumes and grasses) through “cut and carry system” is still contributing

significantly to the nutrition of buffaloes, however, weather extremes limit the use of green fodders

to certain seasons. Thus, during extreme fodder scarcity periods (May to June and November to

December) in Pakistan, buffaloes are completely converted to cereal straws to meet their energy

and protein demands (Khan et al., 2006). Silage production is an important strategy to overcome

the seasonal fodder deficiencies.

The efforts are being done to introduce silage making from different various fodder crops

in Pakistan. The most ideal silage crop is maize (Bolsen et al, 1996; Evitayani et al, 2004),

however, oats, sorghum, barley, millet, mott, and jumbo herbs can also be ensiled (Khan et al.,

2006). The buffalo calves less than one year of age raised on different feeding systems showed

range of growth performances. Burque et al. (2008) investigated the growth performance in buffalo

calves raised on urea treated wheat straw based TMR and reported average daily gain of 0.76 kg.

Anjum and Cheema (2016) reported 0.69 kg average daily gain in buffalo calves raised on millet

silage. Afzal et al. (2009) raised buffalo calves from weaning to one year of age on oat and maize

fodder under cut and carry system and reported an average daily gain of 0.43 kg.

The current experiment was planned to check the growth performance of Nili Ravi buffalo

calves raised on three different silage crops (maize, sorghum and oats) during summer in

subtropical region of Pakistan.

Experiment 4

78

MATERIALS AND METHODS

Experimental animals and housing

Twenty four young male calves of Nili-Ravi buffalo with an average age of 7.0±1 months

(mean ± SD) and weight 110±08kg were raised during summer of 2013, at Buffalo Research

Institute, Pattoki, District Kasur, Pakistan. All experimental calves were kept in a shed as a tie stall

system and individually fed. Free excess of water to each calf was provided by individual water

tubs. Before the start of the experiment, all calves were vaccinated against Foot and Mouth and

Black Quarter disease and dewormed for both external and internal parasites.

Treatments and experimental design

The twenty four calves were randomly divided into 4 groups under completely randomized

design and assigned one of the following four treatments with six calves in each treatment.

MF = Ad libitum maize fodder + concentrates @ 0.5% of body weight

MS = Ad libitum maize silage + concentrates @ 0.5% of body weight

SS = Ad libitum sorghum silage + concentrates @ 0.5% of body weight

OS = Ad libitum oat silage + concentrates @ 0.5% of body weight

Experimental diets were offered twice daily at 9:00 h and 18:00 h. Feed refusals were measured after

24 h before every morning feeding. The feeding trial lasted for ninety days including fifteen days of

adaptation period starting from 14th May and terminated on 8th August, 2013. The composition of

roughages has been shown in Table 6.1.

Parameters studied

Feed offered, and refusal of each animal was recorded daily to calculate DM intake (DMI).

The weight was recorded for each calf on fortnightly basis to estimate the average daily weight gain.

Feed efficiency was also measured (weight gain/ DMI).

Experiment 4

79

Digestibility trial

During the last week of study period, a digestibility trial was carried out for three days to

estimate the dry matter digestibility (DMD) of DM, CP, and NDF. A calf of each group was

randomly selected for the trial, and intake and total feces were collected and weighed daily.

Laboratory Analyses

A composite fecal grab of 30 g in triplicate was dried in a hot-air oven (Memmert,

Beschickung-Loading Model 100-800, Germany) at 60°C for 72 hours (for DM%), then ground

through hammer mill (Wiley laboratory Mill, Standard Model No. 2, Arthur H. Thomas Company,

USA) making particle size of about 0.5 to1 mm for proximate analysis. The feed and fecal samples

were analyzed for CP (AOAC, 1990), NDF, ADF (Van Soest et al., 1991) and ME at Animal

Nutrition Laboratory, Ravi campus UVAS. The gross energy of the silage samples was determined

through the IKA C-2000 Bomb Calorimeter, while metabolizable energy (M.E) was calculated as

63% of the gross energy (Mandal et al., 2003).

Statistical Analysis

The data thus collected were analyzed through ANOVA under CRD using SAS 9.1. The

difference among means was tested through DMRT (Duncan, 1955). The significance level was set

at P≤0.05.

RESULTS

Nutrients intake and weight gain

Dry matter, CP, NDF and ADF intake has been presented in Table 2. Dry mater intake was

significantly higher in calves of group MF (3.38±0.04), however similar between MS (2.44±0.03)

and SS (2.43±0.02) but lower in group OS (2.24±0.02kg/day). While a similar trend was observed

for CP. NDF (neutral detergent fiber) and ADF (acid detergent fiber) intake were also significantly

higher in group MF, but similar between groups all other groups (Table 2). The daily weight gain in

Experiment 4

80

treatment MF, MS, SS, and OS was 373.21±30.28, 389.52±9.09, 407.60±41.50, and 353.26±36.45

respectively. Although, these values were numerically different however the weight gain was

statistically similar among the treatment groups.

Dry matter digestibility and feed efficiency

Dry matter digestibility revealed non-significant (P > 0.05) result between the groups.

Numerically low digestibility of DM was observed in MF (53.94±0.82) and OS (51.16±2.86) while

higher in MS (54.77±1.34) and SS (54.44±0.16). Similar pattern existed for CP and NDF

digestibility among the groups (Table 3).

Numerically higher feed efficiency was observed in calves fed MS diet (0.167) followed

by SS (0.160), OS (0.157) and MF (0.110). The calves consumed more feed on MF diet to gain

one kg of body weight, followed by SS and OS and MF diet.

DISCUSSION

Nutrients intake and weight gain

The lower DM intake in silage diets as compared to fodder in present study was in

agreement to Sarwar et al. (2005) who reported higher DM intake in berseem fodder as compared

to berseem silage. The lower DM intake in silage diets could be due to the lower pH of silages

(Ruiz, et al., 1992). The higher NDF intake for fodder diet was also in line with Sarwar et al.

(2005) who found higher NDF intake in berseem fodder as compared to berseem silage. The higher

CP and NDF intake in MF diet in current study could be the results of higher DM intake from

fodder and the fodder diet had higher CP and NDF content as shown in results. The weight gain

in our study was comparable to Afzal et al (2009) who reported an average daily weight gain of

about 0.43 kg. The numerically lowest weight gain in oat silage diet could be due the limited

supply of protein as in oat silage diet the daily protein intake was lowest. Also similar weight gain

in MS and SS diets could be attributed to the similar nutrient intake. More DM intake and

Experiment 4

81

numerically lower weight gain in MF diet compared to MS and SS could be the result of higher

NDF content in MF diet.

Dry matter digestibility and feed efficiency

The findings of apparent dry matter digestibility in present study were similar to the results

of Khan et al. (2006) who reported that DM, CP and NDF digestibility were similar in lactating

buffaloes fed either oat grass or oat grass silage diets. In contrast with these findings Torotich

(1992) reported a decreased in DM and NDF digestibility of silage based diets and attributed this

decrease to low pH of rumen in silage fed diets leading to decline the growth of fiber consuming

bacteria in the rumen. Khorasani et al. (1993) also reported significantly higher DMD of fodder

due to higher concentration of soluble carbohydrates and lower lignin content in the fodder than

that of its silage. Numerically low digestibility of fodder diet might be due to poor quality of local

summer fodder as well as harsh environmental condition which lead to increased lignification in

the crop.

The calves consumed more feed on MF diet to gain one kg of body weight and calves on

MS diet performed best and were highly efficient to gain one kg weight. The results of our study

are similar to Khan et al., (2006) who described that the higher intake increased the rate of passage

of digesta and therefore reduced the digestibility of feed. Whenever the digestibility of dry matter

is less, the amount of absorbed material from gut will be less, and excretion of materials from the

gastrointestinal tract will be high and this can affect daily gain and feed efficiency. Also reduced

feed efficiency in calves fed on maize fodder could be attributed to poor quality local summer

fodder as well as harsh environmental condition which lead to increased lignification in the fodder

crops.

Experiment 4

82

LITERATURE CITED

Anjum MI, Cheema AU. 2016. Feeding value of millet harvested as silage or hay fed to buffalo

calves supplemented with concentrate on growth performance and nutrient digestibility

Pakistan J. Zool., vol. 48(1), pp. 101-105.

Afzal M, Anwar M, Mirza MA, Andrabi SMH. 2009. Comparison of growth rate of male buffalo

calves under open grazing and stall feeding system. Pakistan Journal of Nutrition 8 (2):

187-188.

AOAC: Official Methods of Analysis. 1990. 15th

Ed., Association of Official Analytical Chemists,

Arlington, Virginia, USA.

Burque AR, Abdullah M, Babar ME. , Javed K, Nawaz H. 2008. Effect of urea feeding on feed

intake and performance of male buffalo calves. J. Anim. Pl. Sci. 18(1).

Bolsen KK, Ashbell G, Weinberg ZG. 1996. Silage fermentation and silage additives: Review.

Asian-Aust. J. Anim. Sci. 9:483-489.

Duncan DB. 1955. Multiple range and F-Tests. Biometrics, 11, 1-42.

Evitayani, Warly L, Fariani A, Ichinohe T, Fujihara T. 2004. Seasonal changes in nutritive value

of some grass species in west Sumatra, Indonesia. Asian-Aust. J. Anim. Sci. 17:1663-

1668.

Khan MA, Iqbal Z, Sarwar M, Nisa M, Khan MS, Lee WS, Lee HJ, Kim HS. 2006a. Urea Treated

Corncobs Ensiled with or without Additives for Buffaloes: Ruminal Characteristics,

Digestibility and Nitrogen Metabolism. Asian-Aust. J. Anim. Sci. 19:705-712.

Khan MA, Sarwar M, Nisa M, Khan MS, Bhatti SA, Iqbal Z, Lee WS, Lee HJ, Kim HS, Ki KS.

2006b. Feeding Value of Urea Treated Wheat Straw Ensiled with or without Acidified

Molasses in Nili-Ravi Buffaloes. Asian-Aust. J. Anim. Sci. 19:645-650.

Experiment 4

83

Khorasani GR, Jedel PE, Helm JH, Kennelly JJ. 1997. Influence of stage of maturity on yield

components and chemical composition of cereal grain silages. J. Anim. Sci. (77): 259–

267.

Mandal AB, Paual SS, Pathak NN. 2003. Nutrient requirements and feeding of buffaloes and

cattle. Published by Int. Book Distributing Co. Charbagh, Lucknow, India. P-23.

Ruiz TM, Sanchez WK, Straples CR, Sollenberger LE. 1992. Comparison of “Mott” dwarf

elephant grass silage and corn silage for lactating dairy cows. J. Dairy Sci., 75: 533-540.

Sarwar M, Khan MA, Nisa M, Touqir NA. 2005. Influence of berseem and Lucerne silages on

feed intake, nutrient digestibility and milk yield in lactating nili buffaloes. Asian-

Australasian. J. Anim. Sci.18 (4): 475-478. doi: http://dx.doi.org/10.5713/ajas.

Steel GD, Torrie JH, Dickey DA. 1997. Principles and procedures of statistics, 3rd ed., Mc

Graw-Hill, New York.

Torotich MJ. 1992. Minimizing the loss of ammonia during urea treatment of wheat straw for

growing calves. MSc Thesis, Haryana Agri. Univ. Hissar, India.

Van-Soest PJ, Robertson HB, Lewis BA. 1991. Method of dietary fiber and non-starch

polysaccharides determination in relation to animal material. J. Dairy. Sci. (74): 3583-

3591.

Experiment 4

84

Table 6.1. Nutritional composition of different roughages fed to buffalo calves

Roughages

Nutrients Maize fodder

(MF)

Maize silage

(MS)

Sorghum silage

(SS)

Oat silage

(OS)

DM% 20.46 27.02 25.24 28.73

CP% 6.23 7.76 6.59 6.38

NDF% 64.65 62.48 61.17 63.54

ADF% 34.78 26.09 29.70 34.92

GE cal/kg 3820 3998 3916 3929

pH - 3.66 3.80 3.71

Table 6.2. Nutrients intake and weight gain of buffalo calves fed different roughages

Parameters Roughages

Maize fodder

(Mean ± SE)

Maize silage

(Mean ± SE)

Sorghum silage

(Mean ± SE)

Oat silage

(Mean ± SE)

DMI (kg/day) 3.38±0.04a 2.44±0.03b 2.43±0.02b 2.24±0.02c

CPI(kg/day) 0.242±0.01a 0.188±0.0.004b 0.160±0.003c 0.138±0.002d

NDF (kg/day) 2.27±0.06a 1.57±0.034b 1.68±0.033b 1.65±0.027b

ADF (kg/day) 1.184±0.031a 0.638±0.013b 0.893±0.017b 0.895±0.014c

Weight gain

(g/day)

373.21±30.28 389.52±9.09 407.60±41.50 353.26±36.45

Means in a same row with different superscripts are significantly different (P<0.05).

Experiment 4

85

Table 6.3. Nutrients digestibility of different roughages fed to buffalo calves

Roughages

Nutrients Maize fodder

(Mean ± SE)

Maize silage

(Mean ± SE)

Sorghum silage

(Mean ± SE)

Oat silage

(Mean ± SE) P.V

DMD (%) 53.94±0.82 54.77±1.34 54.44±0.16 51.16±2.86 0.421

CPD (%) 66.15±0.63 67.51±0.46 66.40±0.74 65.05±0.37 0.066

NDF (%) 50.25±0.69 51.77±0.54 51.56±1.64 49.92±2.22 0.752

Feed efficiency

(%)

0.110

(11%)

0.167

(16.7%)

0.160

(16%)

0.157

(15.7%)

Mean with different superscript in a row differ significantly (P<0.05)

86

CHAPTER 7

EXPERIMENT-5

Impact of replacement of maize fodder with maize silage in total mixed ration on nutrient

intake, digestibility and production performance of early lactating Nili-Ravi buffaloes

ABSTRACT

Sixteen early-lactating (40±10 days in milk), primiparous Nili-Ravi buffaloes with similar milk

production were selected and randomly divided into 4 treatment groups with 4 animals in each

group. Four iso-nitrogenous and iso-caloric experimental TMRs were formulated with 60:40 ratio

of roughage to concentrates on DM basis. The roughage part (60%) of four TMRs comprising

maize fodder (MF) and maize silage (MS) of different ratio. Treatment A had 60% maize fodder

+ 0.0% maize silage, Treatment B 40% maize fodder + 20% maize silage, treatment C 20% maize

fodder + 40% maize silage and treatment D had 0.0% maize fodder + 60% maize silage on DM

basis respectively. Whereas, concentrate was similar for all TMRs. Dry matter intake (DMI)

significantly decreased with increasing addition level of maize silage in TMR. The highest DMI

was recorded in group A (control, 17.99±0.06) followed by B (17.81±0.06), C (17.68±0.06) and

D (16.22±0.07). However, a similar trend was also observed for CP, NDF and ADF intake in all.

DM digestibility significantly higher in control (64.49±0.54) followed by B, C and lowest in group

D (60.17±0.75) respectively. While crude protein (CP) digestibility was also exposed a significant

trend amongst group A with B, C and D, but non-significant among group B, C and D. Furthermore

a similar pattern for neutral detergent fiber (NDF) and acid detergent fiber (ADF) digestibility

were perceived among the groups. Milk yield was significantly (P<0.05) different stand as

8.97±0.12, 8.61±0.09, 8.45±0.10and 8.31±0.07 respectively for group A, B, C and D. Similarly,

milk composition of buffaloes fed on different TMR also linearly different among the groups. Milk

fat % increasing with increasing silage inclusion in TMR the highest fat% was observed in group

C and D (7.18±0.14 and 7.17±0.12) while lowest in A and B (6.76±0.10 and 6.76±0.11). Similarly

Experiment 5

87

solid not fat (SNF), protein, lactose and ash were also significantly (P<0.05) higher in silage based

TMR as compare to fodder base.

INTRODUCTION

Buffalo (Bubalus bubalis) is the major dairy animal in Pakistan contributing about 67% of

total milk production in the country (Afzal et al., 2007). Most of the animals are raised in rural

areas in small herds (< 3 animals) and are fed seasonal fodders or cereal crop leftovers. In peri

urban areas the dairy buffalo is kept for commercial purposes and raised on wheat straw based

TMR along with small quantity of fresh fodder. The seasonal fodder production poses a challenge

for dairy farmers to feed their animals when fodder supply is limited especially during summer

(May - July) and winter (November – January) months (Rasool et al., 1996). Silage is a viable

solution to ensure its supply during those lean periods (Khan et al., 2011).

Despite benefits of silages, it has been reported that the DM intake was lower in silage fed

dairy cows due to the fermentation products causing lower pH in silages (Ruiz, et al. 1992). Also

DM content of silages might also affect the DM intake in dairy cattle (Yahaya et al., 2004). In

dairy buffaloes, the silage feeding has not been extensively researched (Sarwar et al., 2005). Under

such scenario, a study was needed to explore the effect of fodder replacement with silage in

lactating dairy buffaloes.

The objective of present study was to investigate the effect of maize fodder replacement

with maize silage on productive performance in lactating dairy buffaloes.

MATERIALS AND METHODS

Experimental animals and housing

The experiment was carried out at Buffalo Research Institute, Pattoki, District Kasur,

Pakistan during winter months (from 20th October, 2013 and terminated on 20 Jan 2014). Sixteen

early-lactating (40±10 days in milk), primiparous Nili-Ravi buffaloes with similar milk production

were selected and randomly divided into 4 treatment groups with 4 animals in each group. All

Experiment 5

88

experimental animals were kept in a shed as a tie stall system and individually fed. Free excess of

water to each animal was provided by individual water tub. Before the start of the experiment, all

animals were vaccinated against and dewormed according to the farm protocol.

Treatments diets

Four iso-nitrogenous and iso-caloric experimental TMR were formulated with 60:40 ratio

of roughage to concentrates on DM basis. The experimental diets differed in roughage source.

Treatment 1 had 60 % maize fodder (MF) as roughage in the TMR, treatment 2 had 40% maize

fodder + 20% maize silage (MF40), treatment 3 had 20% maize fodder + 40% maize silage

(MF20), and treatment 4 had 0.0% maize fodder + 60% maize silage (MF0) on DM basis, whereas,

concentrate was similar for all TMRs. The chemical composition of all 4 diets has been presented

in Table 1. The treatment diets were offered for 90 days.

Variables recorded

The measured quantity of roughage and concentrate of respective TMR was mixed daily

and offered twice a day (morning and evening) at adlibitum (10% feed refusal) on individual

feeding basis and orts were measured in the morning during the whole study period. All

experimental animals in each treatment had free access to clean, fresh water throughout the day.

Nutrients intake included DM, CP, NDF and ADF of each animal was recorded daily, whereas,

body weights of all animals was taken monthly. Milk production was recorded daily (morning and

evening) and milk samples were collected weekly for fat, protein and ash composition analysis.

Digestibility trial

Three days digestibility trial was carried out in the last week of experiment as described by

Khan et al. (2004). One animal from each group was randomly selected for the trial. Random fecal

samples were taken twice a day so that a sample for every 3 h interval within 24 h (8 samples)

between meals in the morning and the afternoon was obtained (Sarwar et al., 1991). For each

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collection, 50 g of sample was weighed and composited to form one sample per animal.

Laboratory analyses

Feed, orats and fecal samples were dried in the oven at 60 ˚C for 72 h and analyzed for

DM%, percent digestibility and proximate analysis. The samples were analyzed for CP (AOAC,

1990), NDF, ADF (Van Soest et al., 1991) and ME at Animal Nutrition Laboratory, Ravi campus

UVAS. The gross energy of the feed samples was determined through the IKA C-2000 Bomb

Calorimeter, while metabolizable energy (M.E) was calculated as 63% of the gross energy (Mandal

et al., 2003). The milk was analyzed with Lactoscan-S Milk Analyzer (50 W, Milkotronic Ltd.,

Bulgaria) to determine the fat, not solid fat, protein, lactose and ash in Operation Quality

Laboratory BRI, Pattoki.

Statistical analysis

The collected data were analyzed by one-way ANOVA techniques under completely

randomized design using SAS software 9.1.3 Portable. Differences between treatments means

were tested through the Duncan multiple range test (Duncan, 1955).

RESULTS

Nutrients intake

Nutrients intake of 4 different TMRs fed to early lactating Nili Ravi buffaloes has been in

Table 2. Dry matter intake significantly decreased with increasing inclusion level of maize silage

in TMR (p>0.05). The highest dry matter intake was recorded in MF (17.99±0.06) followed by

MF40 (17.81±0.06), MF20 (17.68±0.06) and MF0 (16.22±0.07). A similar trend was observed for

CP, NDF and ADF intake.

Digestibility

Dry matter digestibility was significantly higher in lactating buffalo fed on MF TMR

(64.49±0.54) followed by MF40 (62.07±0.20), MF20 (60.81±0.45) and MF0 (60.17±0.75) (p<

0.05; Table 2). Dry matter digestibility decreased with increased level of silage in TMR. The CP

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digestibility was significantly (P<0.05) higher in MF diet compared to all others (Table 2). All the

three silage diets had similar CP digestibility. Similarly, highest NDF digestibility (53.07±0.50)

was observed in MF group followed by MF40 (51.78±0.29), MF20 (50.93±0.34) and MF0

(50.67±0.18).

Milk yield and milk composition

The results showed that with increasing percentage of silage in TMR, milk yield was

negatively affected in dairy buffaloes (Table 3). The average milk yield was significantly (P<0.05)

higher in MF (8.97±0.12) followed by MF40 (8.61±0.09), MF20 (8.45±0.10) and MF0 (8.31±0.07)

(Table3). The milk composition showed contrary trends as that of milk yield. Milk fat % increased

with increasing silage inclusion in TMR. The highest fat% was observed in group MF20 and MF0

(7.18±0.14 and 7.17±0.12), and lowest in MF and MF40 groups (6.76±0.10 and 6.76±0.11)

accordingly (Table 3). Similarly SNF, protein, lactose and ash were also significantly (P<0.05)

higher in silage based TMR groups as compared to only fodder group (Table 3).

DISCUSSION

Nutrients intake

The decreased in DM intake in silage groups in present study was in line with Sarwar et al.

(2005) and who reported a decrease in DM intake in berseem silage fed lactating buffaloes. They

attributed the decrease in DM intake to the presence of fermentation products in silage. Similarly,

Touqir et al. (2009) compared the replacement of Jambo grass (Sorghum bicolour Sorghum

sudanefe) with Jambo grass silage in Nili-Ravi buffaloes and found a decrease in DM intake of

silage fed animals. Likewise, Ruiz et al. (1992) explained that low pH due to the fermentation

products in silages might cause the decrease in DM intake. The replacement of maize fodder with

maize silage had the same effect on DM intake as that of other silage crops. Decreased CP intake

with increasing silage ratio was in agreement to what has been previously reported (Sarwar et al.,

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91

2005; Touqir et al., 2009). The degradation of CP to non-protein nitrogen during ensiling process

might explain the decreased CP intake in silage fed animals (Ruiz et al. (1992).

The higher NDF and ADF intake in high fodder groups (MF and MF40) in present study

was similar to previous findings (Sarwar et al., 2005; Khan et al., 2006; Touqir et al., 2009). As

the high silage diets had relatively high fiber in our study that might have compromised DM intake

due to filling affect and also lowered the NDF intake. The fiber content in silage based TMR in

our experiment was higher than fodder because fodder was harvested at early stages and for silages,

crop was harvested at optimum stage of maturity having DM of about 30 %. Decreased ruminal

pH in silage fed diets might also have affected the degradation of NDF due to higher lactic acid

contents and thus reduced NDF intake.

Digestibility

The current results of lower DM, CP, and NDF digestibility in silage based TMR were in

line with previous studies (Sarwar et al., 2005; Touqir et al., 2009). The higher digestibility in

fodder based diets could be due to higher concentration of soluble carbohydrates and lower lignin

content in the fodder than that of its silage (Khorasani et al. (1993). Also, low pH of silage due to

lactic acid contents, decreased the ruminal pH and thus reduced hemicellulose and cellulose

digestibility by depressing the growth of cellulolytic bacteria in the rumen of silage fed animal

(Sarwar et al., 2005).

The DM, CP, NDF digestibility values were also in range as reported previously. Sarwar

et al. (2005) reported digestibility of DM as 64.8 and 62% and CP as 72 and 71.5% for berseem

fodder and berseem silage, respectively. Khan et al. (2006) reported 71.3 and 71.1% CP

digestibility for oat fodders and oat silage, respectively. Likewise Touqir et al. (2009) reported

71.3 and 71.13% CP digestibility for jumbo grass and jumbo silage respectively. However, the

current findings showed slightly higher DM digestibility than that of Khan et al (2006) and Touqir

et al. (2009). The different fodders in their diets could explain that difference. However, it was

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92

evident from these findings that CP digestibility was more consistent in buffaloes that DM and

NDF irrespective of fodder types.

Milk yield and milk composition

Higher milk yield in high fodder compared to high silage based diet was in agreement to

Sarwar et al. (2005) who reported higher milk yield in berseem fodder compared to berseem silage

diets. Likewise, Touqir et al. (2009) reported a higher milk yield in sorghum grass than sorghum

silage. Higher milk yield could be due to higher DM intake in fodder based diets. Increased milk

fat % in our study was similar to Touqir et al. (2009) who found higher milk fat in silage based

diets. The acetate is a major end product of lactate fermentation, the conversion of lactic acid

content of silage-based diets to acetate might have improved the milk fat contents. (Chamberlain

and Roberston., 1992; Man and Wiktorsson., 2001). Increased protein content in current study

was in line with Givens and Rulquin (2004) who found increase in milk protein concentration in

silage fed diets and attributed that to efficient microbial protein synthesis.

Conclusion

The results of present study revealed that fresh corn fodder can be successfully replaced

with corn silage in lactating buffaloes. Although dry matter intake was lower in silage based diets

that lead to slightly reduced milk yield but the total solids were higher.

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LITERATURE CITED

Afzal M, Anwar M, Mirza MA .2007. Some factors affecting milk yield and lactation length in

Nili-Ravi buffaloes. Pakistan. Vet. J. (27): 113-117.

AOAC: Official Methods of Analysis. 1990. 15th

Ed., Association of Official Analytical Chemists,

Arlington, Virginia, USA.

Chamberlain DG, Roberston S. 1992. The effects of the addition of various enzyme mixtures on

the fermentation of perennial rye grass silage and on its nutritional value for milk

production in dairy cows. Anim. Feed Sci. Technol. 37:257.

Duncan DB. 1955. Multiple range and F-Tests. Biometrics, 11, 1-42.

Givens DI and Rulquin H 2004. Utilisation by ruminants of nitrogen compounds in silage-based

diets. Animal Feed Science and Technology 114, 1–18.

Khan SH, Azim A, Sarwar M, Khan AG .2011. Effect of maturity on comparative nutritive value

and Fermentation characteristics of maize, sorghum and millet silages. Pak. J. Bot., 43(6):

2967-2970.

Khan MA, Sarwar M, Nisa M, Iqbal Z, Khan MS, Lee WS, Lee HJ, Kim HS .2006. Chemical

composition, in situ digestion kinetics and feeding value of oat grass (avena sativa) ensiled

with molasses for Nili-Ravi Buffaloes. Asian-Aust. J. Anim. Sci. 19 (8): 1127-1133.

Khan MA, Sarwar M, Nisa M, Khan MS. 2004. Feeding value of urea treated corncobs ensiled

with or without enzose (corn dextrose) for lactating crossbred cows. Asian-Aust. J. Anim.

Sci. 17:1093-1097.

Khorasani GR, Jedel PE, Helm JH, Kennelly JJ. 1997. Influence of stage of maturity on yield

components and chemical composition of cereal grain silages. J. Anim. Sci. (77): 259–

267.

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Mandal AB, Paual SS, Pathak NN. 2003. Nutrient requirements and feeding of buffaloes and

cattle. Published by Int. Book Distributing Co. Charbagh, Lucknow, India. P-23.

Rasool S, G. Ahmad G, Abdullah M. 1996. Fodder conservation, crop residues and by products in

livestock production system. Proceeding of national conference on the improvement

production and utilization of fodder crops in Pakistan. March 21-27, Pakistan Agriculture

Research Coubcil / FAO, Islamabad, Pakistan, p. 176-186.

Ruiz TM, Sanchez WK, Straples CR, Sollenberger LE. 1992. Comparison of “Mott” dwarf

elephant grass silage and corn silage for lactating dairy cows. J. Dairy Sci., 75: 533-540.

Sarwar M, Khan MA, Nisa M, Touqir NA. 2005. Influence of berseem and lucerne silages on feed

intake, nutrient digestibility and milk yield in lactating Nili buffaloes. Asian-Aust. J. Anim.

Sci. Vol. 18, No. 4: 475-478.

Steel GD, Torrie JH, Dickey DA. 1997. Principles and procedures of statistics, 3rd ed., Mc Graw-

Hill, New York.

Sarwar M, Firkins JL, Estridge ML. 1991. Effect of replacing neutral detergent fiber of forage with

soy hulls and corn gluten feed for dairy heifers. J. Dairy Sci. 74:1006-1015.

Tauqir NA, Sarwar M, Jabbar MA, Mahmood S. 2009. Nutritive value of jumbo grass (sorghum

bicolour sorghum sudanefe) silage in lactating Nili-Ravi buffaloes. Pakistan. Vet. J. 29(1):

5-10.

Man NV, Wiktorsson H. 2001. Cassava tops ensiled with or without molasses as additive effect

on quality, feed intake and digestibility by heifers. Asian-Aust. J. Anim. Sci. 14:624-631.

Van-Soest PJ, Robertson HB, Lewis BA. 1991. Method of dietary fiber and non-starch

polysaccharides determination in relation to animal material. J. Dairy. Sci. (74): 3583-

3591.

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Yahaya MS, Goto M, Yimiti W, Smerjai B, Kawamoto Y. 2004. Additives effects of fermented

juice of epiphytic lactic acid bacteria and acetic acid on silo fermentation and ruminal

degradability of tropical elephant grass. J Anim Vet Adv. 3:115–121.

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Table 7.1. Ingredients and chemical composition of four experimental TMR (DM basis)

Ingredients% TMR

A(control) B C D

Maize fodder 60.00 40.00 20.00 ------

Maize silage ------- 20.00 40.00 60.00

Cotton seed cake 04.00 04.00 04.00 04.00

Canola meal 10.00 10.00 10.00 10.00

Soybean meal 15.00 15.00 15.00 15.00

Maize grain 3.00 3.00 3.00 3.00

Rice polishing 5.00 5.00 5.00 5.00

Molasses 2.00 2.00 2.00 2.00

Minerals Mixture 1.00 1.00 1.00 1.00

Total % 100 100 100 100

Chemical composition%

DM 25.59 28.45 32.04 35.62

CP 16.93 16.69 16.45 16.21

NDF 38.70 39.10 40.35 41.70

ADF 22.78 23.06 24.02 25.47

GE.cal/gm 4037 3986 3968 3942

TMRs: A) contained 60% maize fodder, B) 40% maize fodder + 20% maize silage, C) 20% maize

fodder + 40% maize silage and D) 60% maize silage respectively. The remaining 40% DM in each

TMR was supplied by the concentrates

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Table 7.2. Nutrient intake (kg) and digestibility of maize fodder and its silage based TMR

Means within each row followed by different superscripts are significantly different (P<0.05)

Nutrients Intake

Groups

A(control) B C D

DM 17.99±0.06a 17.81±0.06b 17.68±0.06b 16.22±0.07c

CP 3.23±0.01a 3.19±0.02b 3.15±0.01c 2.86±0.01d

NDF 9.48±0.06a 9.44±0.04a 9.23±0.08b 8.75±0.04c

ADF 4.58±0.01a 4.51±0.02b 4.35±0.02c 4.27±0.02d

Digestibility%

DMD 64.49±0.54a

62.07±0.20b 60.81±0.45bc 60.17±0.75d

CPD

72.90±0.32a

70.89±0.28b

70.70±0.27b

70.11±0.27b

NDF

53.07±0.50a

51.78±0.29b

50.93±0.34bc

50.67±0.18c

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Table 7.3. Milk yield and milk composition of four experimental groups

Yield kg /day

Groups

A(control) B C D

Milk yield 8.97±0.12a 8.61±0.09b 8.45±0.10bc 8.31±0.07c

Composition%

Fat 6.76±0.10b 6.76±0.11b 7.18±0.14a 7.17±0.12a

SNF 10.15±0.05b 10.32±0.07a 10.37±0.05a 10.35±0.05a

Protein 3.94±0.02b 4.05±0.03a 4.06±0.02a 4.06±0.03a

Lactose 5.31±0.03b 5.45±0.04a 5.47±0.02a 5.44±0.03a

Ash 0.83±0.003b 0.84±0.006a 0.84±0.003a 0.84±0.004a

Means within each row having different superscripts are significantly different (P<0.05).

99

CHAPTER 8

SUMMARY

Silage production is at initial stages to be a part of animal agriculture in Pakistan. The lack

of research on silage making and its benefits for livestock production under local conditions is an

important factor for slow propagation of silage in our country. Under such scenario a multi-step

study was conducted. At first, the effect of proper maturity stage for harvesting different fodders

was investigated, and then the effects of silo type and silage additives were assessed on silage

quality. In last part of the study the feeding trials were conducted on growing calves and lactating

buffalo to evaluate the effect of silage feeding on growth and milk production respectively.

In all the three fodders i.e. oats, maize and sorghum, the full bloom stage for harvesting

produced the best results regarding silage quality and fermentation characteristics. Although the

trench silo produced best results regarding fermentation characteristics and silage quality, the

expected operational cost and dry matter losses during face management for trench silo would

make it harder for farmers to adopt. Under such circumstances, for long term use the bunkers

would be a good choice for silage making with comparable silage quality as that of trench silo.

Silage inoculants certainly improved the silage quality and it is highly recommended to use such

additives for silage making and these additives are not that costly. Buffalo calves raised on fresh

corn fodder and three different silages showed similar daily weight gain. Further growth trials on

buffalo calves with varying levels of concentrate feeding along with silage are suggested to

investigate silage feeding in calves. Corn silage fed lactating buffaloes had lower dry matter intake

and total milk yield, but higher total solids as compared to fresh fodder feeding. The future studies

of silage feeding compared to different inclusion levels of fresh and dry roughage sources would

add further to explore the economic implications of silage feeding.