EFFECT OF FEEDING DECORTICA TED COTT ONSEED CAKE …...Ishtiyak sir, Vijay sir, Nazam sir, Sanjay...

EFFECT OF FEEDING DECORTICATED COTTONSEED

CAKE ON GROWTH PERFORMANCE IN

CROSSBRED CALVES

THESIS SUBMITTED TO THE

NATIONAL DAIRY RESEARCH INSTITUTE, KARNAL

(DEEMED UNIVERSITY)

IN PARTIAL FULFILMENT OF THE REQUIREMENT

FOR THE AWARD OF THE DEGREE OF

MASTER OF VETERINARY SCIENCE

IN

ANIMAL NUTRITION

BYREKHA MOURYA

(B.V.Sc. & A.H.)

DAIRY CATTLE NUTRITION DIVISION

NATIONAL DAIRY RESEARCH INSTITUTE

(I.C.A.R.)

KARNAL-132001 (HARYANA), INDIA

2013Regn. No. 2091109

DEDICATED DEDICATED DEDICATED DEDICATED

TO MYTO MYTO MYTO MY

FAMILY ,FAMILY ,FAMILY ,FAMILY ,

PROFESSION PROFESSION PROFESSION PROFESSION

&&&&

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ACKNOWLEDGEMENTACKNOWLEDGEMENTACKNOWLEDGEMENTACKNOWLEDGEMENT

Words have always failed to express the magnitude of feelings, particularly

when it comes to acknowledging the help rendered.

I express my deep sense of gratitude to my preceptor Dr. J.P. Sehgal,

Principal Scientist, Dairy Cattle Nutrition Division, NDRI, Karnal, for his benevolent

and sagacious guidance, constant encouragement, invaluable suggestions, keen

interest, sympathetic understanding and morale boost up during the entire course of

this research work. I feel lucky to get a mentor like Sehgal sir who helped me at every

time when I was in need of help and guidance of any and at any time. My

indebtedness is reserved for his goodwill and patience during entire period of my

research work. I am really thankful to him for everything.

I express my profound sense of gratitude to all the members of my advisory

committee namely, Dr. S.S. Kundu, Principal Scientist& Head DCN Division; Dr. A.

K. Tyagi, Principle Scientist, DCN Division; Dr. Mahendra Singh, Principle

Scientist, DCP Division; Dr. Shiv Prasad, Principle Scientist, LPM Section; for

extending their incisive guidance, valuable suggestions and constructive ideas at

every stage of this endeavour.

I am also thankful to Dr. Madhu Mohini, Principle Scientist, DCN Division;

and Dr. Anjali Aggarwal, Principle Scientist, DCP Division for their immense help

in Invitro and haematological study.

My sincere thanks are also to Dr. A.K. Srivastava, Director, NDRI, Karnal for

providing necessary facilities for carrying out this study and ICAR for financial

assistance in the form of Junior Research Fellowship during my Master’s

programme.

I am also very much thankful to all my respected seniors Drs. Ajaz sir,

Ishtiyak sir, Vijay sir, Nazam sir, Sanjay Sawant sir, Sukhjinder Jeet sir ,Madhu

Suman mam, Anjila mam, Sonali mam, Farukh sir, Sakendra sir, Umesh sir, who

helped me in every way when I was in need of their help. I wish to express my cordial

appreciation and special thanks for their kind support and affection, which has

enabled me to complete this work.

No words are enough to thank my batchmates Drs. Lalesh, Shailesh, Vimlesh,

Mithlesh, Puneet, Vikrantjeet, Suraj, Nana, Bhawana, Abhilasha, Bisitha and Tho for

their whole hearted bolster during the entire period in this institute.

The support and encouragement of my roommate Anjali, my seniors in

Alaknanda hostel Drs. Rupal mam, Bharti mam and Shilpi mam, whose cooperation

made my stay at the institute a pleasurable.

The help rendered by Mr. H. K. Meenaji and Mr. Rajpalji (Uncle ji) during my

research work is duly acknowledged. A formal line of appreciation is not sufficient to

express my gratitude and indebtedness to them.

I am also thankful to Virenderji, Surjeet, Lalit, Sandeep and other worker of

cattle yard for their cooperation and help during entire the experiment.

Words would never be able to fathom the depth of my feeling for my loving

husband and all family members, who displayed enormous strengths, courage and

preservance in helping me to complete my degree.

At last but not least, I record my sincere thanks to all beloved and respected

people, who helped me, could not find separate mention.

Date: July, 2013 (Rekha Mourya)

CONTENTS

Chapter Title Page no.

1 Introduction 1-4

2 Review of literature 5-17

2.1 Cotton crop production in world 5

2.2 Feeding value of cottonseed cake 6

2.3 Effect of cottonseed or cottonseed cake on feed intake

and digestibility of nutrients

8

2.4 Effect of decorticated cottonseed cake on animal growth

performance

9

2.5 Effect of cottonseed cake on lactation performance 10

2.6 Effect of cottonseed / cottonseed cake on blood

biochemical profile

11

2.7 Cottonseed cake as a substitute for soybean meal 12

2.8 Cottonseed cake vis-à-vis gossypol (anti nutritional

factor )

14

2.9 Plasma gossypol in lactating dairy cattle fed cottonseed

and cottonseed cake

16

3 Material and method 18-34

3.1 Proximate analysis of different feed ingredients 18

3.2 Estimation of cell wall constituents 22

3.3 In vitro (1st stage) technique (Tilly and Terry, 1963) 24

3.4 In vivo study 26

3.5 Metabolic trial 29

3.6 Haematological parameters 31

3.7 Stastical analysis 35

4 Results and discussion 35-56

4.1 Proximate composition and cell wall components of

different ingredients concentrate mixture

35

4.2 Invitro studies 49

4.3 Growth study 41

4.4 Effect of partial replacement of soybean meal with

cottonseed cake or decorticated cottonseed cake on

43

Chapter Title Page no.

voluntary feed intake, productive performance and

percent feed efficiency

4.5 Nutrient intake and nutrient utilization 46

4.6 Nutritive evaluation in terms of percent digestible crude

protein (DCP) and total percent total digestible nutrients

(% TDN)

49

4.7 Nitrogen balance 50

4.8 Blood biochemical profile 52

4.9 Economics of feeding 54

5 Summary and conclusion 57-59

6 Bibliography i - xi

LIST OF TABLES

Table

No. Title

Page

No.

3.1 Description of animals 28

4.1 Percent ingredient composition of experimental

concentrate mixtures fed to female crossbred calves 36

4.2 Percent chemical composition and cell wall contents of

different ingredients used in concentrate mixture (on

Dm basis)

37

4.3 Percent chemical composition of concentrate mixtures

SBM ‘C’, ‘T-1’( replacing 10 parts of soybean meal with

cottonseed cake expeller), ‘T-2’ (replacing 10 parts of

soybean meal with decorticated cottonseed cake

solvent extracted), green oats and wheat straw

38

4.4 In vitro degradability of different cakes after incubation

at 48 hrs. 40

4.4(a) In vitro degradability of different concentrate mixtures

after incubation at 48 hrs. 41

4.5 Effect of partial replacement of soybean meal (SBM)

with cottonseed cake (CSC) or decorticated cottonseed

cake (DCSC) on fortnightly body weight changes (kg)

in female crossbred calves

42

4.6 Productive performance of female crossbred calves fed


cake (DCSC) as a partial replacement of soybean meal

(SBM) in complete feed mixtures

45



cake (DCSC) on nutrient intake (kg) in female

crossbred calves (during metabolic trial)

47

Table

No. Title

Page

No.



cake (DCSC) on digestibility coefficient (%) of various

nutrient of different complete feed mixture in female

crossbred calves

49



cake (DCSC) on nutritive evaluation of rations in terms

of % DCP and % TDN in female crossbred calves

50



cake (DCSC) on nitrogen balance in female crossbred

calves

51



cake (DCSC) on blood biochemical profile in female

crossbred calves

53



cake (DCSC) on economics of feeding

55

LIST OF FIGURES

Figure

No. Title

After

Page No.

1 Effect of partial replacement of soybean meal (SBM)

with cottonseed cake (CSC) or decorticated

cottonseed cake (DCSC) on fortnightly body weight

changes in female crossbred calves

42

LIST OF PHOTOS

Photo

No. Title

A/Page

No.

1 Decorticated cottonseeds cake 27

2 Animal during Metabolic trial 29

LIST OF ABBREVIATIONS

% = Percentage

* = multiplication

@ = At the rate

˚C = degree Celsius

ADF = Acid detergent fibre

< = Less then

≤ = less or equal

> = More than

µl = Microlitre

a.m. = Ante Meridian

ad lib = ad libitum

ANOVA = Analysis of varience

ARC = Agriculture research council

BW = Body weight

CaCl2 = Calcium chloride

cm = centimeter

CP = Crude protein

CSC = cottonseed cake

DCSC = decorticated cottonseed cake

SBM = soybean meal

d = days

CPD = Crude protein digestibility

CRD = Complete randomized design

DCP = Digestible Crude protein

DM = Dry matter

DMD = Dry matter digestibility

DMI = Dry matter intake

EE = Ether extract

et. al. = Co-workers

Fig. = Figure (s)

g = Gram (s)

h/hr = Hour

HCl = Hydrochloric acid

i.e. = That is

IVDCPDR = In vitro ruminal crude protein digestibility

IVDMDR = In vitro ruminal dry matter digestibility

IVOMDR = In vitro ruminal organic matter digestibility

KCl = Potassium chloride

KF = Karan Fries

kg = Kilogram

kg/d = Kilogram per day

lbs = Pound

Lt. = litre (s)

m = meter (s)

mg = Milligram (s)

MgCl2.7H2O = Magnesium chloride hydrated

mm = millimeter (s)

MY = Milk Yield

NDF = Neutral detergent fibre

Na2CO3 = Sodium carbonate

Na2HPO4 = Disodium hydrogen phosphate

NaHCO3 = Sodium bicarbonate

NDRI = National Dairy Research Institute

No. = Number

NRC = National Research Council

N.S. = Not significant

NFE = Nitrogen free extract

OMD = Organic matter digestibility

p.m. = Post Meridian

RDP = Rumen degradable protein

RUP = Rumen Undegradable protein

SE = Standard error

Sr. No. = Serial number

SEM = Standard Error mean

TDN = Total digestible nutrients

TMR = Total mixed ration

UDP = Undegradable protein

v/v = Volume by volume

VFA = Volatile fatty acid

Yr = Year

ABSTRACT

To observed the effects of partial replacement of soybean (de-oiled) meal with cottonseed expeller or decorticated cottonseed cake solvent extracted on voluntary feed intake, digestibility of nutrients, nutritive evaluation of the feeds; growth of calves; blood profile and % feed efficiency; 18 karan fries female calves of similar age (8-9 months) and body weight (114-115 kg) were randomly divided into three groups of each and allotted diets ‘C’, ‘T-1’ or ‘T-2’. The diet ‘C’ having 15% soybean meal. Which was replaced by 10 parts with cottonseed cake expeller in ‘T-1’ diet or 10 parts by decorticated cottonseed cake solvent extracted in ‘T-2’ diet. All the crossbred female calves were fed individually rations (wheat straw: concentrate 50:50) along with 10kg oats green fodder/ animal/ day. Body weight was recorded fortnightly and blood samples collected at monthly intervals. An in vitro rumen degradability study of different diets was also undertaken to determine the rumen degradability of DM, OM and CP. The in vitro rumen dry matter degradability (IVDMDR) (%) significantly higher (P≤0.05) in ‘C’ (71.41±1.26) followed by ‘T-1’ (67.92±1.10) and ‘T-2’ (65.05±0.93) group. The in vitro rumen organic matter degradability (IVOMDR) (%) was significantly higher (P≤0.05) in ‘C’ (67.03±0.85) and lowest in ‘T-2’ (55.03±0.91) group. In vitro rumen crude protein degradability (IVCPDR) was significantly lower (P≤0.05) in ‘T-2’ (56.54±1.25) followed by ‘T-1’ (59.04±1.41) and ‘C’ (65.74±1.73). The dry matter intake (kg/day) was 4.14±0.40, 3.83±0.44 and 4.04±0.39 in ‘C’, ‘T-1’ and ‘T-2’ respectively. The dry matter intake (kg/100kg BW) was 2.50±0.42, 2.35±0.45 and 2.71±0.57 in ‘C’, ‘T-1’ and ‘T-2’ respectively, which did not differ significantly. The average daily gains were observed to be significantly higher (P≤0.05) in group ‘T-2’ (750±51g/day) than group ‘C’ (634±61 g/day) and group ‘T-1’ (583±37 g/day). The digestibility of ether extract (%) was significantly (P≤0.05) higher for ‘T-1’ (87.16±1.04) and ‘T-2’ (86.39±0.93) than ‘C’ (79.34±1.43). Whereas, digestibility of DM, OM, CP, CF, ADF, NDF and NFE did not differ significantly in all three groups. Nitrogen balance was significantly higher in ‘T-2’ (37.30±5.61) and ‘C’ (30.95±2.61) than ‘T-1’ (28.78±1.14), all animals were found in positive nitrogen balance. There was no significant difference in nitrogen intake and nitrogen retention in all three groups. Blood urea nitrogen was significantly higher (P≤0.05) in ‘T-2’ (18.08±0.64) than ‘C’ (14.77±0.61) and ‘T-1’ (14.67±0.58) while blood glucose, total protein, albumin, globulin, albumin: globulin ratio and haemoglobin not differ significantly in all three groups. The cost for per kg body weight gain was(Rs) 73.65, 69.38 and 56.78 in ‘C’, ‘T-1’ and ‘T-2’ respectively. It can be concluded that soybean meal de-oiled can be replaced 10 parts with decorticated cottonseed cake solvent extracted to have higher growth rate; nutritive digestibility; nutritive evaluation and % feed efficiency in growing female calves.

प ( )

( ( ) , ,

, , औ %

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CHAPTER CHAPTER CHAPTER CHAPTER –––– 1111

INTRODUCTIONINTRODUCTIONINTRODUCTIONINTRODUCTION

1 IntroductionIntroductionIntroductionIntroduction

INTRODUCTION

Due to population explosion in India, there will be additional needs of

oils and fats besides grains. As there is a big shortage in oil seeds due to low

production and also export of quality oil seeds; the oil mills are extracting

complete oil from seeds or cakes through solvent extraction. Thus compared

to the oil seed cake solvent extracted cake are made available in plenty for

animals. In these days conventional protein supplements, such as full fat

soya, soya oil cake, soybean meal, mustard oil cake and groundnut cake tend

to increase the cost of animal production due to either their being transported

from other regions or being considered as export commodity in India. Thus,

the replacement of traditional ingredients of feeds with alternatives should be

attempted to lower the maintenance and production costs. The rations

formulated from alternative ingredients with small quantity of traditional

supplements must be efficient and economical, and should offer the same

performance as that of other balanced compounded feeds.

Cotton is very important cash crop in Punjab, Haryana, Maharashtra

and Gujarat. After removing the cotton fibre, the seeds are available for oil

production. Because of use of cottonseed oil in vegetable ghee preparation,

the expeller pressed cotton seed or decorticated cottonseed cake is becoming

available for animals. Hence, it will be in the interest of the animal nutritionists

to know their nutritive value for ruminants. As also now a days due to

increased prices and less availability of soybean meal dairy farmers are

seeking a suitable and viable alternate protein supplement to soybean meal or

full fat soybean (extruded). Among the alternatives cotton seed meal (solvent

extracted) is one which contain 38-44% crude protein, though it contain less

protein than soybean meal de-oiled (47%) but matches to soybean full fat as a

protein source (38%).

Several oil seeds have been investigated for this purpose (Njike, 1977;

Nzekwe and Olomu, 1982; Bamgbose, 1995). Cottonseed cake is a traditional

protein and energy supplement in dairy cattle ration and as an alternative to

soybean meal contains 68.31% total digestible nutrients (TDN), 38.03% crude

protein (CP), 34.92% neutral detergent fibre and 20.37% non-fibrous


carbohydrates (Valadares Filho et al., 2006). It also has reasonable

palatability and is available at a lower cost than soybean meal, thereby

making its use in animal feed a viable alternative (Lana, 2000).Whole

Cottonseed, cottonseed meal (decorticated) and cottonseed meal

(undecorticated) all are excellent protein ingredients. Whole cottonseed is

high in protein, energy and fiber. It is also a good, though, variable source of

thiamine but a poor source of carotene (Obioha, 1992).

It is a good source of phosphorus and vitamin E and is easily available

in considerable quantity in India. Supplementation of crushed cottonseed in

the ration of cow’s decreases heat increment lowers body temperature and

reduces heat stress of summer (Moody, 1962). In cotton growing regions,

whole cottonseed (WCS) is readily available and in some cases, it becomes

economical to use small-scale expander-expeller equipment to remove a

portion of the oil, which creates a by-product feed ingredient referred to as

expanded-expelled cottonseed. Approximately half the oil is removed during

this process. The expander-expeller process exposes the cottonseed protein

to some heat to make it undegradable in rumen. Excessive heat treatment of

feed stuffs may decrease proteolysis by denaturing the protein, thus blocking

reactive sites of amino acids through microbial proteolytic enzymes. Heat has

been used to decrease protein degradation in the rumen and to increase the

supply of dietary protein to the duodenum. Cottonseed cake, a by-product

from oil industry, is high in protein, and is a good source of crude fiber and

phosphorus. However, both contain gossypol, a polyphenolic bi-naphthyl

dialdehyde that can produce toxic effects in milch animals at higher levels of

inclusion in feeds. However, ruminants have the ability to detoxify large

amounts of gossypol within the rumen. Diets containing up to 25 percent

whole cottonseed have been reported to be safe for consumption by cattle.

Prior to 1980, gossypol was seldom mentioned in reports after feeding

cottonseed or cottonseed meal to mature cattle. It was known that young

cattle, prior to the development of a fully functional rumen, were susceptible to

gossypol poisoning, compared to mature cattle; because, during digestion of

cottonseed and cottonseed meal, gossypol remained bound in the rumen so

that it was mostly unavailable. However, Lindsey et al. (1980) demonstrated

that the ability of the rumen to detoxify gossypol exceeded when very high


intakes of free gossypol were consumed by mature cows. The unique physical

characteristics of cottonseed, along with its better nutritional value, have

resulted in its being extensively used in ruminant’s diets, but concerns about

gossypol have made producers reluctant to use cottonseed or cottonseed

cake in their diets. Cottonseed cakes contain three types of fibre - acid

detergent fibre, neutral detergent fibre and crude fibre. The types of fibre

determine the overall quality. Cottonseed meal is a good protein source for

ruminants. Cottonseed meal has shown promises as a plant protein substitute

to the conventional ones, especially, soya bean meal and groundnut cake.

According to Njike (1977), cottonseed meal is the residue obtained after the

extraction of oil from cottonseed and the product is ground resulting in flakes.

It is produced in commercial quantities in Nigeria and it’s relatively cheap.

Bamgbose (1995) observed that cottonseed cake contains 36.15% protein,

19.96% fibre, 14.42% fat, while biological value was 51.0% compared to 61.0

and 73.0% for defatted and full-fat extruded soybean meals. The protein of

cottonseed meal was found to be low in cystine, methionine and lysine

(Nzekwe and Olomu, 1982). It is palatable with a nutritive value (for dehulled

meals) slightly lower (85-90 %) than that of soybean meal. It is among the

least expensive sources of protein in some region. It is for instance the main

source of protein for livestock in the cotton growing belt of India (NDDB,

2012).

Cottonseed meal has relatively low rumen degradability and is

therefore a good source of by-pass protein that is useful in rations for dairy

cows. Sehgal and Makkar (1994) reported that cottonseed cake

(undecorticated) has higher degradable protein (47.0%) compared to

groundnut cake. Cottonseed meal can replace other oilseed meals (soybean,

rapeseed, sunflower and groundnut) without affecting milk yield and

composition. When supplementing highly digestible forages such as maize

silage, cottonseed meal (expeller) can replace soybean meal without any

detrimental effect on DM intake and milk yield (Coppock et al., 1987).

Now because presently sufficient quantity of high protein decorticated solvent

extracted cottonseed meal is available after the extraction of complete oil from

the cottonseeds by solvent extracted plant, it will be in the interest of the dairy


farmers to use this product. Hence the present study has been conducted with

following objectives:

• To study the in vitro protein degradability of decorticated cottonseed

cake (DCSC), cottonseed cake, soybean meal, groundnut cake and

other ingredients of the concentrate mixture.

• To study the effect of partial replacement of soybean meal with

decorticated cottonseed cake (Solvent extracted) and cottonseed cake

(Expeller) on nutrient utilization, body weight gain in growing calves

and on blood biochemical profile.

CHAPTER CHAPTER CHAPTER CHAPTER –––– 2222

REVIEW OF REVIEW OF REVIEW OF REVIEW OF LITLITLITLITEEEERRRRAAAATURETURETURETURE

5 Review of LiteratureReview of LiteratureReview of LiteratureReview of Literature

REVIEW OF LITERATURE

2.1 COTTON CROP PRODUCTION IN WORLD

Cotton as a fibre crop is produced all around the world but mostly in the

tropics. It does not grow wild as in early days but grows commercially as a

cash crop. The major producers of cotton are also the major producers of

cottonseeds. China, India, USA, Pakistan, Uzbekistan and Brazil are the

major producers globally. The trade in cottonseed as oil crop is very less

compared to other oilseeds. Of the total global production of 35 million tons,

27 million tons is crushed for oil production and around 8 lakh tons is globally

traded. Other by-products of cotton crop like cottonseed oil and cottonseed oil

cake have their production figures hovering around 4.5 and 16 million tons

respectively. India has the distinction of having the largest area under cotton

cultivation and is the second largest producer of cotton and its derivatives,

having annual production of cottonseeds and cottonseed oil cake around

11.50 and 4.0 million tons, respectively, (USDA, 2011).

Although cotton is cultivated in almost all the states in the country, the

states of Punjab, Haryana, Rajasthan, Gujarat, Maharashtra, Madhya

Pradesh, Andhra Pradesh, Tamil Nadu and Karnataka account for more than

95 percent of the area under cultivation and output. These states are also the

major producers of cottonseeds as this is a major by-product of cotton

crop. Cottonseeds arrive in the market as a by-product of ginning operation

carried on cotton. Cottonseeds are oilseeds those are place 2nd in the list of

largest produced oilseeds in the world. This oil seed has a vast number of

uses as it provides oil for humans, feed for animals, fertilizer for plants,

padding fibre and also used in explosives and computer chip boards.

Solvent extracted decorticated cottonseed meal is normally sold as a

41% protein product but is available as 35% corticated cottonseed cake, 38

%( de-oiled) and 44% decorticated cottonseed protein meals. They contain

over 1% phosphorus and have 70-80% TDN. Cottonseed cakes made from

whole cottonseed contain 18.1 percent acid detergent fibre, 32.3 percent

neutral detergent fibre and 11.4 percent crude fibre. Animals fed with whole


cottonseed or cottonseed cake (undecorticated) would receive substantially

more fibre in their diets.

Cottonseed cake contains minerals such as: calcium, magnesium,

phosphorus, potassium, sodium, sulphur, copper, iron, manganese,

molybdenum and zinc. They also contain some amino acids viz: alanine,

aspartic acid, cystine, glutamic acid, glycine, histidine, lysine, leucine,

arginine, isoleucine, methionine, phenylalanine, proline, serine, threonine,

tryptophan, valine and tyrosine. The minerals provided in cottonseed cakes

cannot meet an animal's mineral nutritional needs. In such cases, the feed

may be combined with other types of grain alfalfa or oats, for instance to

provide proper balance.

Cottonseed cake is an excellent source of protein for a variety of

animal species. Cottonseed cake is often further processed into pellets of

varying size (1/4", 3/8", 1/2", 3/4" and 7/8") depending on the application.

Cottonseed meal can be fed alone or in combination with other plant and

animal protein sources to make a complete balanced ration. The

characteristics of a particular meal are largely determined by the type of oil

extraction process from which the meal was derived - mechanical (screw

press) or expander solvent extraction.

2.2 FEEDING VALUE OF COTTONSEED CAKE

Cottonseed cake (CSC) has been successfully tried as animal fed

ingredient for more than 100 years in areas where cotton production and

processing is prevalent. This is used primarily as a protein supplement for a

variety of beef production operations that include calf creeps and beef cows

feed ingredients. After oil, cottonseed cake is the second most valuable and

most abundant by-product of the crushing process from cottonseed.

The nutrient analysis of cottonseed cake is dependent upon the

process used to extract the cottonseed oil. The standard undecorticated

cottonseed cake contains 25% crude protein on as such fed basis. The crude

fibre level of cottonseed cake is significantly higher (13 vs. 5%) than that of

soybean meal. Consequently, the protein and energy content of cottonseed

cake undecorticated is lower than soybean meal.


According to Coppock (1987), the nutritional protein degradability of

cottonseed cake is similar to that of peanut meal, canola meal, and soybean

meal for lactating dairy cows, and to that of canola and soybean meal for

young calves. However, Sehgal and Makkar (1994) reported a lower protein

degradability of cottonseed cake (60.5%) compared to GNC (76.3%).

From an historical perspective, when Oklahoma researchers (Hibberd

et al., 1987) added increasing levels of cottonseed cake to low-quality native

grass hay diets containing equal amounts of corn. They observed a significant

improvement in digestibility of feed. Several growth trials have supported

these results through comparable performance using either hay or silage

based diets (Brown, 1991).

Several research trials with beef cows have estimated the protein and

energy value of cottonseed cake, relative to other protein sources, under a

variety of dietary conditions. A Louisiana study (Coombs, 1996) evaluated the

effect of self-fed protein supplements during late gestation and early lactation,

with an energy ingredient during the second half of the supplementation

period, on cow body weight changes and subsequent calf performance. Cows

also had ad libitum access to a Bermuda grass hay (9.9% crude protein and

49.6% TDN) throughout the supplementation period. The experimental

supplement evaluated included cottonseed cake with salt (desired daily intake

of 0.7kg), a commercially available high protein (40%), and low protein (20%)

block. The result showed that there was no difference among protein

supplement sources on cow body weight change and weaning weights of

calves and when low-quality native grass hay (4.7% crude protein) was used

as the base diet.

Gonzalez et al. (1988) supplemented fall-calving cows at calving with

1.12kg of cottonseed cake daily. During the first five weeks of lactation, the

control treatment (no protein supplementation) lost more than 100 lbs of body

weight, while the cows supplemented with cottonseed cake gained almost 50

lbs. Hay intake was higher by 33% for control and 110% for the cottonseed

cake supplemented cows during the first five weeks after calving. The

supplemented cows produced more milk with faster weight gain than control

cows. This study showed that small quantities of cottonseed cake efficiently


improved the utilization of low-quality forage and performance of lactating

beef cows.

Florida researchers conducted two trials that evaluated the effects of

supplemental cottonseed cake on the performance of nursing beef calves

(Kunkle et al., 1991). The nursing calves averaged 430 to 560 lbs at the

initiation of the two summer trials. Consumption of the cottonseed cake- salt

supplement averaged 452g per head per day in trial 1 and 345g per head per

day in trial 2. The calves supplemented with cottonseed cake gained 255g

per head per day more in trial 1 and 200g per head per day more in trial 2

compared to control.

2.3 EFFECT OF COTTONSEED OR COTTONSEED CAKE ON FEED

INTAKE AND DIGESTIBILITY OF NUTRIENTS

According to Church (1993), one of the most important factors that can

influence digestibility is the diet composition. According to Van Soest (1994),

Orskov (2000) and McDonald et al. (2002), the factors that could affect the

nutrient digestibility would be feed intake, the proportion and degradability of

the cell wall, the composition of the feed component, feed composition, feed

preparation, protein-to-energy ratio, rate of degradability and factors inherent

to the animal.

Wanapat et al. (1996) found that straw intake was significantly

decreased from 1.25% to 0.88% of live weight as the level of cottonseed cake

was increased from 2-5 kg/d in dairy cows. However, a summary of 18

feeding trials (Coppock et al., 1987) with whole cottonseed showed no

significant reduction (P > 0.05) in dry matter intake when whole cottonseed

was included up to 25% of the ration.

Shirley (1998) reported no difference in DMI when feeding expanded

expelled cottonseed in place of Whole cottonseed. Pena et al. (1986) also did

not found any change in OM intake when feeding, raw, extruded or roasted

cottonseed. Similarly other studies (Saijpaul et al., 2006; Sullivan et al., 1993)

did not find any difference in DMI when feeding whole linted cottonseed and

cracked or ground pima cottonseed. Arajuo et al. (2004) also observed no

influence of the treatments on the nutrient intake.


Van Horn et al. (1979) analysed different sources of protein

(cottonseed meal and soybean meal) and different CP levels in the diet (13.5

and 16.3%) reported an increase in DM intake in both treatments; however,

this increase was more significant when the protein source was cottonseed

cake.

Chaudhary and Raj (2008) divided eighteen Surti buffaloes, in their

early stage of lactation, into three groups of six animals each, and allotted

them to three dietary treatments viz. T1 – zero per cent cottonseed in

concentrate mixture, T2 – 30 per cent cottonseed in concentrate mixture and

T3 – 60 per cent cottonseed in concentrate mixture. Additionally, all groups

were fed on basal roughage consisting of 15 kg green Lucerne and ad libitum

Sorghum Stover. The buffaloes of T3 group consumed significantly (P<0.05)

higher DM, DCP and TDN than T1 group, but there were no significant

differences observed between T2 and T1 groups.

Silva et al. (2009) found no change in the apparent digestibility

coefficient of the nutrients (P>0.05) with the replacement of soybean meal by

cottonseed cake which was probably due to the similarity in the experimental

diets. However, Pina et al. (2006) reported lower digestibility values for DM,

OM, EE, CP and NDF when analysing the use of cottonseed meal compared

to soybean meal. However, these authors used higher levels of these

ingredients in diets provided to pure-bred animals with greater milk

production.

2.4 EFFECT OF DECORTICATED COTTONSEED CAKE ON ANIMAL

GROWTH PERFOMANCE

When the oil is extracted out decorticated cottonseed cake is a high

protein supplement being produced after removing lint and cortex portion from

cottonseed and generally it is solvent extracted cottonseed cake. Also

cottonseed cake is considered to be a natural source of higher rumen

undegradable protein (Sehgal and Makkar, 1994) i.e. 39 percent known to be

rumen by pass protein. Thus this will be a better protein supplement for higher

growth and milk production in ruminants.

Torane et al. (2006) conducted an experiment to see the effect of

different levels and sources of bypass protein with urea treated or untreated


wheat straw on performance of crossbred calves. One balanced concentrate

mixture (CM) with RDP to UDP ratio of 65:35 was compounded and fed to the

calves in control group C with untreated wheat straw ad libitum. Calves in

experimental groups T1 and T2 were fed ad libitum urea treated wheat straw

with decorticated groundnut cake and un-decorticated cottonseed cake having

RDP to UDP ratio of 78:22 and 52:48, respectively.

They found higher average daily gains for calves in group T2 as

compared to C and T1. Feed efficiency of calves in groups T2 and T1 was

superior over group C. The post feeding blood urea nitrogen level in calves of

group T2 was lower as compared to those in C and T1. The cost of composite

ration per kg gain in body weight for calves in groups C, T1 and T2were found

to have economic superiority of T2 over C and T1.

The results revealed that urea treatment of wheat straw improved the

nitrogen content, voluntary feed intake and the digestibility of nutrients.

Feeding of crossbred calves with urea treated wheat straw based ration with

un-decorticated cottonseed cake containing 48 per cent UDP, stimulated the

growth rate to achieve better and economic feed efficiency in early stages of

growth.

Bangani and co-workers (2000) compared the growth performance of

18 Holstein and 20 Jersey heifers fed calf starter containing either cottonseed

oil-cake (CSC) or soybean oil-cake (SBC). The diets were isonitrogenous and

isocaloric and were fed from two weeks of age until two or three months of

age to Holstein or Jersey calves respectively.

They found no differences between calf starter containing CSC or SBC

(P > 0.05) in respect of feed intake, growth rate or per cent efficiency of feed

conversion. Average daily gain of Jersey and Holstein heifer calves was

0.435±0.02 and 0.635±0.03 kg/day respectively. It was concluded that the

most important criterion for inclusion of CSC in calf starter ration is the cost of

cottonseed oil cake relative to that of soybean oil cake, provided that the

cottonseed oil cake used has a gossypol content of less than 200 ppm.


2.5 EFFECT OF COTTONSEED CAKE ON LACTATION

PERFORMANCE

Coppock and Wilks (1991), in a review article, stated that inclusion of

about 15% of whole cottonseed in the diet of lactating cows resulted in a small

increase in milk yield, milk fat percentages and decrease in milk protein

percentages. This is also the level recommended by Arieli (1998).

Wanapat et al. (1996) found that milk production of crossbred Holstein-

Zebu cows fed a low protein basal diet of rice straw and cassava chips was

markedly improved when the cottonseed meal supplement was increased

from 2 to 4 kg/day. This feeding system is an economical attractive alternative

to farmers who traditionally use commercial concentrates.

Dhiman et al. (1999) found an increase in milk and milk protein yields

and a tendency for increase in fat yield for cows fed a full fat extruded

cottonseed in comparison to control that contained no oilseeds. Depressions

in milk protein have also been reported previously with the feeding of whole

cottonseed (Anderson et al., 1979). Bernard and Calhoun (1997) found no

change in milk yield when they compared diets containing 15% of a 50:50

mixture of extruded cottonseed and soybeans.

However, Imaizumi et al. (2002) found a decrease in milk production as

well as fat, lactose and protein content in the milk when soybean meal was

replaced with cottonseed meal in lactating Holsteins.

Chaudhary and Hemraj, (2008) studied lactating performance of Surti

buffaloes in their early stage of lactation. The animals were divided into three

groups and were allotted to three dietary treatments viz. T1 –0%, T2 – 30%

and T3 – 60% cottonseed in concentrate mixtures. They recorded higher milk

yield in T3 as compared to T1 group and there was no difference in milk yield

of T2 and T1 groups. A vasectomised bull was paraded among the

experimental animals daily for finding the buffaloes in heat and those showing

complete oestrus symptom were inseminated. Their results showed that

higher levels of cottonseed in the diet substantially improved the milk yield

and post-partum reproductive performance of buffaloes.


2.6 EFFECT OF COTTONSEEDS/COTTONSEED CAKE ON BLOOD

BIOCHEMICAL PROFILE

The plasma protein level is a function of the hormonal balances,

nutritional status, water balance and the other factors affecting the health. The

level of the serum albumin is dependent upon the nutritional status and its

synthesis is diminished during fasting, malnutrition and poor condition of liver,

while the serum globulin level is related with immune status of the animal

(Jain, 1986).

The intake of gossypol through cottonseed affects the blood

biochemical profile of the animals (Mena et al., 2004).The blood profile of

cattle given cottonseed @1.2-2.0kg/d showed gossypol toxicity with higher

values of haemoglobin, proteins, albumin: globulin ratio, urea inorganic

phosphorus (Mena et al., 2004), aspartate amino transferase, lactate

dehydrogenase, creatine kinase, cholesterol and triglycerides (Coppook et al.,

1994).

Also Saijpaul et al. (2006) reported that levels of haemoglobin,

glucose, cholesterol, creatinine, urea nitrogen, uric acid, chloride, calcium and

phosphorus in the plasma of experimental animals, fed whole linted

cottonseed @ 5% of their milk yield, did not differ significantly from control

group, but there was a significant rise in the values of total proteins, albumin,

and globulin of the treatment group than control. However the values were

within the normal reported range (Kaneko, 1989).

Similarly Colin et al. (1996) reported no significant change in the blood

glucose and uric acid levels in cattle fed whole cottonseed meal for 430 days.

ALP is a membrane bound enzyme that is used to diagnose bone and liver

disorders while increase in ALT and AST are indicator of soft tissue damage

particularly that of liver. Production of ALT, AST and ALP is increased in

response to primary or secondary hepatic cellular disorders. Liver

degenerative changes occur in gossypol toxicity and the serum activities of

ALP, ALT and AST have been reported to be significantly higher due to

gossypol toxicity by feeding of cottonseed cake (Abd-Allah and El-Fadali,

1996).


2.7 COTTONSEED MEAL AS SUBSTITUTE FOR SOYBEAN MEAL

With increase in prices of the soybean meal de-oiled/full fat soybean,

the dairy farmers will like to use alternative/ cheaper protein supplement.

Cotton seed cake expeller or solvent extracted being cheaper will be a good

source of protein supplement.

Ojewola and co-workers (2006) used cottonseed cake as substitute for

soybean meal in broiler ration. For this 6 week feeding trial was conducted,

cottonseed cake was substituted for soybean meal at 0, 25, 50, 75 and 100%

and the diets were respectively designated as diets 1, 2, 3, 4 and 5 in a

completely randomized design.

The results showed that there were no significant differences in the

bird’s mean daily weight gain and feed-to-gain ratio while the mean daily feed

intake was influenced. Birds fed diets 3 and 4 respectively consumed 150.93g

and 153.68g. Birds fed diet 3 gave the highest weight gain while diet 1 was

the least. Diet 5 had the highest percent mean values for nitrogen (81.45),

crude fibre (60.81), ether extract (95.57), ash (66.79) and dry matter (85.72)

retentions while birds fed diet 1 was least for same parameters.

Mineral utilization followed the same trend; diet 5 was the highest

(69.08), (84.72), (71.91), (79.39) for phosphorous, potassium, calcium and

magnesium respectively while diet 1 gave least values for same parameters.

Diet 5 was found to be the cheapest (N 290.01) and savings were significantly

(P<0.05) improved with the use of this diet while diets 1 and 3 had the least

values of N 285.94 and N 302.67 respectively for marginal revenue. Though,

the diets were comparable, diet 5 showed superior indices for nutrient

utilization and economics of production. In conclusion, there was an indication

that iron treated cotton seed meal can serve as a substitute for soybean meal

in broiler diet.

Balogun et al. (1990) studied the effect of undecorticated cottonseed

cake in weaner and growing finishing Large White × Landrace pigs for two

months studies. A digestibility study has done to determine the value of

undecorticated cottonseed cake (UCSC) as a protein supplement replacing

soya bean meal (SBM). They did a feeding experiment with pigs, four lots of

six pigs each were individually fed on isonitrogenous diets containing 0, 10,

20 and 30% undecorticated cottonseed cake. Their result shows that there is


no adverse effect of feeding undecorticated cottonseed cake on either on feed

intake or on weight gain.

In a second growth experiment lasting 35 days, 24 cross-bred pigs

were fed on diets containing 0, 20, 30 and 41.1% undecorticated cottonseed

cake (without soybean meal) in a randomized design involving six individually

fed pigs per treatment. The growth studies showed that 10% was the optimum

level of inclusion of undecorticated cottonseed cake in the diets of pigs.

2.8 COTTONSEED CAKE VIS-À-VIS GOSSYPOL (ANTI NUTRITIONAL

FACTOR)

Gossypol is a pigment found naturally in many Gossypium species

including cotton and is located in glands throughout the plant. Gossypol is

found in free state in the whole seed and is bound to lysine or other

components during processing into cake. Gossypol bound in this way has

generally been considered unavailable to the animal. Animal sensitivity to

gossypol is considerably different between species and classes of animals.

The quantity of free gossypol has been used as a guide by many nutritionists

to make recommendations on feeding of cottonseed products.

Whole cottonseed has been shown to contain from 0.40 to 2.0% free

gossypol. The level of gossypol is affected by species, variety, fertilization,

growing conditions, and insect pressure. The presence of gossypol affords the

plant some protection against predators such as insects, field mice, and

raccoons that might otherwise feed on these plants and/or their seeds

(Boatner, 1948; Berardi and Goldblatt, 1969).

Gossypol exists as two stereoisomers or mirror images of each other,

which are designated as (+) and (-) isomers. The minus or “(-)” isomer has

been shown to be more detrimental biologically within the animal. These

isomers exist in two distinct states: bound and unbound. The unbound form of

this compound has been shown to be most biologically active in the animal.

The bound gossypol is essentially unavailable to the animal, (because these

are chemical determinations) but the possibility for some crossover of

biological activity exists (Calhoun et al., 1995).

Whole cottonseed typically contains 1.5-2.0% gossypol, all in the

unbound form, but levels can vary to as low as 0.4% in some commercial

species. (Calhoun, 1995, Nomeir and Abou-Donia, 1985). Breakdown and


maceration by chewing of this seed by the animal and subsequent exposure

of this gossypol to rumen microorganisms allows a number of deactivation,

binding, and degradation actions to occur that render the gossypol

unavailable to the animal. Due to the nature of the rumen, prolonged

exposure time, and extensive physical and chemical breakdown of the whole

seed, the ruminant is given some practical protection from the compound.

Binding to free epsilon amino nitrogen in the rumen whether as free amino

acids or peptides attaching to microbial cell walls, or binding to available metal

ions such as iron all contribute to the detoxifying action of the rumen.

Gossypol toxicity may be an issue when feeding large amounts of

cottonseed products. Gossypol is toxic to non-ruminants, but ruminants are

more tolerant (Danke et al., 1965). There are reports of signs of gossypol

toxicity in cows consuming 10.1kg/d of cottonseed cake (Lindsey et al.,1980),

but Blackwelder (1997) fed 8.4kg/day of cotton products (cottonseed cake and

whole cottonseed) without observed toxicity.

Calhoun and Holmberg, (1991) have discussed the potential for

gossypol toxicity in cattle consuming cotton products. They believed that free

gossypol was the toxic form of gossypol and as bound gossypol was not

available to ruminants and it was not toxic. Consequently, recommendations

for using cottonseed and cottonseed meal in animal diets were based on free

gossypol (Berardi and Goldblatt, 1980; Calhoun and Holmberg, 1991). Total

gossypol, which includes both free and bound gossypol was seldom

determined.

Rogers and Poore (1995) reported that it was safe to feed higher

levels of free gossypol in the diet when the source of free gossypol was whole

cottonseed instead of cottonseed meal. For example, for mature cows and

bulls the recommended safe levels of free gossypol in the diet were 900 and

1,200 ppm when whole cottonseed was fed compared with 200 and 600 ppm,

respectively, when the source of free gossypol was cottonseed cake. The

higher levels of free gossypol recommended for diets containing cottonseed

reflects the fact that free gossypol in unprocessed whole cottonseed is

extensively bound during digestion in the rumen and is less available than

free gossypol in cottonseed cake.


Further Calhoun, (1995) and Mena et al. (2001) concluded that the

plasma gossypol response to free gossypol intake was 3.8 times higher when

the source of free gossypol was from cottonseed cake indicating higher

availability of free gossypol in cottonseed cake compared to cottonseeds.

Santos et al. (2002) in Holstein steers observed that cracked whole

cottonseed increased plasma concentrations of total gossypol, although total

gossypol intake measured as g/d or as mg/kg of live weight per day did not

differ.

However, in studies with cattle and sheep, in which different sources of

free gossypol (gossypol acetic acid, cottonseed and cottonseed cake

processed by different methods) were fed, it was observed that plasma and

tissue levels of gossypol and toxicity often were not correlated with either the

free gossypol content of the feed or free gossypol intakes (Calhoun et al.,

1990; Calk, 1992; Calhoun and Wan, 1995; Wan et al., 1995). These studies

demonstrate that free gossypol is of limited value for making

recommendations about levels of cottonseed and cottonseed cake that can be

fed safely to ruminants.

The official methods for the estimation of free and total gossypol are

spectrophotometric procedures based on the reaction of aniline with gossypol

to form dianilino gossypol (AOCS, 1985). Although widely used, this

procedure is only applicable to the determination of gossypol in cottonseed,

cottonseed cake and untreated cottonseed meals. They are unsuitable for the

determination of gossypol in mixed feeds because of interferences with other

components of feeds (Pons, 1977) and are not selective. They measure not

only gossypol, but also gossypol analogues and gossypol derivatives, with an

available aldehyde group that are soluble under the conditions of the

methods. The most serious limitation of spectrophotometric procedures is

they do not determine the (+) and (–) isomers of gossypol.

The gossypol molecule exhibits optical activity because of stearic

hindrance to rotation about the internaphthyl bond (Matlin et al., 1988). High

performance liquid chromatography (HPLC) procedures also have been used

to measure free and total gossypol in cottonseed and cottonseed meal (Hron

et al., 1990). They are selective for gossypol and are 50 to100 times more

sensitive than the official methods (Pons 1977; Hron et al., 1990).


2.9 PLASMA GOSSYPOL IN LACTATING DAIRY CATTLE FED

COTTONSEED AND COTTONSEED CAKE

Prieto et al. (2003) studied on lactating Holstein cows which were fed

diets with increasing levels of cracked Pima cottonseed to determine its

effects on plasma gossypol concentrations as well as milk yield and

composition and dry matter (DM) intake in a short-term study. All diets

contained 12.8% cottonseed, 43.5% concentrate, and 43.7% chopped alfalfa

hay on a DM basis. The proportion of whole Upland cottonseed to cracked

Pima cottonseed in the four diets was 100:0, 67:33, 33:67, and 0:100. All

periods were 35 d. Upland and cracked Pima cottonseed contained 0.64 and

1.00% total gossypol (DM) with 41 and 52% of gossypol as the (−) isomer,

respectively.

Total plasma gossypol concentrations increased linearly with

increasing proportions (100:0, 67:33, 33:67, and 0:100) of cracked Pima

cottonseed in the diet for primiparous (4.4, 6.0, 7.7, and 8.9 µg/ml) and

multiparous (4.3, 7.3, 9.7, and 11.4 µg/ml) cows, respectively. This indicates

the importance of animal variation when relating plasma gossypol levels with

gossypol intake.

They found no difference in Milk yield, as well as its components and

DM intake, were not affected by increasing dietary inclusion levels of cracked

Pima cottonseed up to 8.6% of DM intake for either primiparous or

multiparous cows, even though plasma gossypol concentrations increased

sharply over this dietary inclusion range. Although the highest dietary

inclusion level of Pima cottonseed (i.e.,12.8%) numerically depressed

performance of cows of both parties, these differences failed to reach

statistical significance in these short-term trials with few cows.

Wen-Ju Zhang and co-worker studied on Upland and Pima two main

varieties of cottonseed (CS), and Pima CS is considered nutritionally superior

to Upland CS because of its higher fat and protein content. Pima CS contains

more gossypol and a higher proportion of the (−) isomer than Upland CS.

They found that DM intake of dairy cattle is not altered when WCS is

included at up to 25% of the diet. The DM intake response to the inclusion of

WCS in the diet is a function of both climatic and dietary factors.

CHAPTER CHAPTER CHAPTER CHAPTER –––– 3 3 3 3

MATERIALMATERIALMATERIALMATERIALSSSS AND METHODAND METHODAND METHODAND METHODSSSS

18 Materials & Methods

MATERIALS AND METHODS

In vitro and In vivo experiments were carried out to study the rumen

dry matter, organic matter and protein degradability of feed incorporating

cottonseed cake (undecorticated) and decorticated solvent extracted

cottonseed cake replacing 10 part of soybean meal. Also in vivo feed intake,

nutrient utilization, feed conversion ratio and blood parameters in growing

Karan fries calves fed with concentrate mixture have different composition.

The materials and methods used for this study are given below.

3.1 PROXIMATE ANALYSIS OF DIFFERENT FEED INGREDIENTS

(AOAC, 2005)

3.1.1 Dry matter (DM)

Apparatus: Aluminium moisture cup, hot air oven, desicator, electronic

balance, tong, spatula

Procedure

A known quantity of ground sample (about 10 -50 g) was taken in a

pre-weighed moisture cup. The cup was placed in hot air oven at 100 ± 50C

for 24 h.

The loss in moisture content after drying was estimated and DM was

calculated as follows

(Wt. of moisture cup + sample after drying – Wt. of moisture cup) X 100

DM (%) = ------------------------------------------------------------------------------------

Wt. of fresh sample

3.1.2 Total ash (TA)

Apparatus: Silica crucible, hot plate, muffles furnace, electronic weighing

balance and tong.

Procedure

A known quantity of sample (about 2.5 - 5 g) was taken in pre-weighed

silica crucible. After charring the sample on heater (till the smoke


disappeared), the crucible was kept in muffle furnace for ignition at 550°C for

4 h. The crucible was removed on cooling and kept in a desicator and

weighed again to find out weight of ash. The ash content was calculated as

given below:

(Wt. of crucible + ash after cooling - Wt. of crucible) X 100

Total ash (%) =

Wt. of sample (g)

Organic matter (OM)

Procedure

OM was determined by subtracting the total ash content from 100.

OM (%) = 100 – total ash (%)

3.1.3 Crude protein (CP)

Apparatus: Kjeldahl flasks, digestion tubes, digestion unit, Kjeldahl distillation

apparatus, Erlenmeyer flasks, titration assembly/burette.

Reagents: Digestion mixture (Na2SO4 and CuSO4 in the ratio of 9:1), 40%

NaOH solution (400 g NaOH pellets dissolved in distilled water and volume

made to 1000 ml), concentrated H2SO4 (98% purity and specific gravity 1.84)),

2% boric acid indicator solution (20 g boric acid dissolved to 1 L and added

with 10 ml 0.2% bromocresol green and 20ml 0.1% methyl red indicators),

N/100 H2SO4 solution.

Procedure

Crude protein (Nx6.25) was determined by Kjeldahl’s technique. A

known quantity of dry ground sample was taken in digestion tube and

digested with concentrated sulphuric acid and 2-3 g of digestion mixture

(potassium sulphate and copper sulphate in the ratio of 9:1) till the solution

become colourless. After digestion, the contents were cooled and volume was

made to 100 ml in a volumetric flask. A suitable volume of digested sample

was transferred in to the micro-kjeldhal distillation apparatus and an

approximate quantity of 40% sodium hydroxide solution was added to make


the content alkaline. A 100 ml distillate was collected in a conical flask

containing 2% boric acid solution with mixed indicators .The distillate was then

titrated against standard sulphuric acid solution (N/100).

Calculation

0.0014× Titre volume × Normality × Volume made N (g/100 g sample) = ---------------------------------------------------------------- x 100 (Aliquot taken× Sample taken)

The crude protein (%) of sample was calculated by multiplying the N content

with the factor 6.25. This was based on the principle that all the proteins

contain 16% nitrogen.

3.1.4 Ether extract (EE)

Apparatus: Soxhlet’s extraction apparatus, oil flask, thimble, hot air oven,

desicator, weighing balance

Reagent: Petroleum ether (boiling point 40-60oC)

Procedure

A known quantity of ground sample (about 3 g) was taken in a cellulose

thimble and extracted for 8 hours with petroleum ether in Soxhlet’s extraction

apparatus attached to a pre weighed oil flask. The oil flask was removed and

after evaporating the excess of ether, it was dried overnight in a hot air oven

(100±5°C). The flask was cooled in a desicator and weighed to a constant

weight. The difference between two weights gave the amount of ether extract

in the sample.

Calculation

(Wt. of oil flask with ether extract – Wt. of empty oil flask)

EE (%) = --------------------------------------------------------------------------- X 100

Wt. of sample

3.1.5 Crude fibre:-

Apparatus :- Hot plate, hot air oven, muffle furnace, spout less beaker,


condensation unit for flask (round bottom flask as condenser), muslin cloth

(24 threads each Warf and weft per sq. inch), wash bottle, steel spatula,

Buchner funnel with suction arrangements ( suction pump attached to swan

tap at one end and other end to side neck of conical flask having funnel).

Reagent :-Sulphuric acid solution 0.255N or 1.25g H2SO4/100ml, Sodium

hydroxide solution 0.313N or 1.25g of NaOH/100ml, antifoam agent (e.g. octyl

alcohol or silicon), ethyl alcohol at 95%(v/v), acetone.

Procedure

A measured amount of moisture and fat free sample was taken in

spout less beaker of 1 litre capacity previously marked to 200 ml. Then, 25 ml

of 10% sulphuric acid (w/w) was added and volume was made up 200 ml with

water from the sides of the beaker to have 1.25% acid solution (w/w) in the

beaker. The contents of beaker were successively refluxed for 30 minutes.

The beaker contents then filtered through muslin cloth with the help of

Buchner funnel with suction arrangements. After repeated hot water washing,

the residue left on muslin cloth was transferred to the same spout less beaker

with smooth steel spatula, followed by little washing of muslin cloth.

Again 25 ml 10% NaOH (w/v) was added and volume was made up to

200 ml with water to have 1.25% alkali solution. Contents were refluxed for 30

minutes and filtered through the muslin cloth again and washed with hot

water. The residue left on muslin cloth was transferred to clean silica crucible

with the help of steel spatula and little water washing. The content was dried

in hot air oven at 100±2°C and weighed. Silica containing dried residue was

kept in a muffle furnace at 550 °C ±5 °C for 4 hrs, for ashing and again

weighed next day. The crude fibre content of sample was estimated as

follows.

Crude fibre (g/100 g sample or crude fibre %) = (a-b)/w x100

Where,

a= weight (g) of silica basin plus oven dried sample

b= weight (g) of silica basin plus ash


w=weight of sample

3.2 ESTIMATION OF CELL WALL CONSTITUENTS

The fraction of cell wall constituents such as NDF, ADF were estimated (Van

Soest et al., 1991).

3.2.1 Neutral detergent fibre (NDF)

Apparatus: Spout less beaker, sintered glass crucible (G-1), vacuum pump,

hot air oven, muffles furnace, electronic balance, and desicator.

Reagents: Neutral detergent solution (NDS), amylase solution, acetone

Neutral detergent solution (NDS)

Sodium lauryl sulphate - 30.00 g

Disodium ethylene diamino tetra acetate (EDTA) - 18.61 g

Sodium borate decahydrate - 6.81 g

Disodium hydrogen phosphate (anhydrous) - 4.56 g

Triethylene glycol - 10 ml

Distilled water - to make volume 1litre

EDTA and sodium borate decahydrate were put together in a large

beaker with some distilled water and heated on hot plate until dissolved.

Similarly, sodium lauryl sulphate was dissolved in distilled water and

triethylene glycol was added to it. The solution of sodium lauryl sulphate and

triethylene glycol was added to the previous solution. Disodium hydrogen

phosphate was taken in another beaker and some amount of distilled water

was added and the contents were heated until dissolved. Then, it was added

to solution containing other ingredients and volume was made up to one litre

with distilled water.


Amylase solution: Dissolve 2 gm α–amylase heat resistant enzyme in 90 ml

water, filter through Whatman 54 paper, stored at 5°C.

Procedure

Approximately 1 g sample was taken in spout less beaker of 1 L

capacity. To this, 100 ml NDS and 0.5g of sodium sulphite were added. The

contents of spout less beaker were refluxed for half an hour. Thirty minutes

after onset of boiling, beaker was removed and 2 ml of enzyme solution was

added. One hour after initial boiling, the contents of beaker were filtered

through pre-weighed 50 ml sintered glass crucible (G-I) using oil-free vacuum

pump. The contents were washed repeatedly with hot boiling water and then

acetone to remove all salts. The sintered crucible containing residue was

dried in hot air oven (100±5°C) overnight, cooled and weighed to a constant

value. Then ashing was done and crucible along with ash was weighed again.

The NDF (ash free) was calculated as follows:

NDF (%) = (Wt. of crucible with residue – wt. of crucible with residual ash) ×100

Wt. of sample taken

3.2.2 Acid detergent fibre (ADF)

Apparatus: Spout less beaker, sintered glass crucible, vacuum pump, hot air

oven, electronic balance and desicator.

Reagents: Acid detergent solution (ADS), acetone, hot boiling water.

Acid detergent solution (ADS): 20 g cetyl trimethyl ammonium bromide

(CTAB) was dissolved in one lit of 1 N H2SO4.

Procedure

Approximately 1 g of sample was taken in a spout less beaker of 1 L

capacity. To this, 100 ml ADS was added and the contents were refluxed for

exactly 1 hour. After refluxing, the residue was filtered through pre-weighed

sintered glass crucible G-I using vacuum pump and washed with hot water 2-

3 times followed by acetone to remove all salts. The sintered crucible


containing residue was dried in hot air oven (100 ± 5°C) and weighed again.

The ADF was calculated as follows:

(Weight of crucible+ fibre) – Weight of crucible

ADF (%) = ---------------------------------------------------------------------- ×100

Weight of sample on DM basis

3.2.3 Hemicellulose

Hemicellulose was soluble in ADS and there by calculated by

subtraction of ADF from NDF as follows:

Hemicellulose (%) = NDF (%) – ADF (%)

3.3 In vitro (First stage) technique (Tilly and Terry, 1963)

The control and experimental feeds were analyzed for the estimation of

rumen dry matter, organic matter and rumen protein degradability.

Reagents

a) McDougall’s buffer solution-

Sr.No Reagents Quantity

1 NaHCO3 49.0 g

2 Na2 HPO4. 18.6 g

3 KCl 28.5 g

4 NaCl 23.5 g

5 CaCl2 2.0 g

6 MgCl2.7H2O 6.0 g

i) Forty nine(49.0) g of NaHCO3 ,18.6 g Na2HPO4 were dissolved in

approximately 800 ml of water,100 ml of chloride solution containing

28.5 g KCl,23.5 g NaCl,6.0 g MgCl2.7H2O and 2.0 g CaCl2 per litre

were added and the mixture made to 1 litre. The solution was

thoroughly saturated with CO2 at 380C until it became clear.


ii) Strained rumen liquor: Rumen liquor was collected from fistulated

animal maintained on standard diet using plastic tube attached through

a filter flask to a vacuum pump.

iii) Immediately after removal, the rumen fluid was strained through four

layers of muslin cloth into a flask. CO2 was passed into the flask to

displace air from above the rumen liquor and it was kept at 38-390C

until required.

Procedure

i) 0.5 g of mill ground sample passed through 0.8 mm sieve and oven

dried at 1000C was placed into already numbered centrifuge tube.

ii) 40 ml of buffer solution was added, followed by 10 ml of the strained

rumen liquor in each tube. The mixture was stirred, gassed with CO2,

kept at 390C. CO2 was passed through each tube for maintaining

anaerobic condition for 48 hrs.

iii) The tube was sealed with a rubber cork fitted with Bunsen gas release

valve and then placed in an incubator at 390C for 48hr.The tubes were

shaken gently 3 or 4 times, a day.

iv) One blank tube, containing only rumen liquor and buffer, and at least two

tubes containing standard samples of known digestibility were included

in each batch of experimental tubes.

v) The pH was maintained during incubation within the limits 6.7-6.9.

Appropriate adjustments were made with 1N Na2CO3.

vi) After 48 hrs. of first incubation period, the bacterial activity was stopped

by adding 1 ml of 5% HgCl2 and 2ml of 1N Na2CO3 are also added to

improve sedimentation.

vii) The tubes were centrifuged at 1800 × g at 10C.

viii) The supernatant from the centrifuge tube was poured off and residue

was transferred to a pre-weighed crucible with minimum amount of

water.


ix) The crucibles plus the residues were dried at 1000C, cooled in a

desicator and weighed. The organic matter residues were obtained

after ashing the crucible plus the residues. Crude protein contents were

also estimated in the residues to find out the percent protein

degradability.

Calculations-

DMD%=

Sample dry matter -

×100

OMD% =

Sample organic matter -

×100

CPD% =

Sample crude protein -

×100

3.4 IN VIVO STUDY

3.4.1 Location of the farm

The experiment was conducted in the cattle yard of NDRI campus,

Karnal situated at an altitude of 250 meter above mean sea level, latitude and

longitude position being 29o 42” N and 79o54” E, respectively. The maximum

ambient temperature in summer goes up to 45oC and minimum temperature

in winter comes down to about 4oC with a diurnal variation to the order of 15-

20oC. The average annual rainfall is 700 mm, most of which is received from

early July to mid September.

3.4.2 Animals and their feeding

Eighteen growing female crossbred calves (Avg. age 8-9 months)

were taken from the institute herd. The animals were divided into three groups


of six each on the basis of their body weights, and age. All the animals were

kept on a measured amount of concentrate mixture, wheat straw and green

fodder @ 10 kg/animal/day to meet their nutrient requirements which were

calculated as per recommendations of NRC (2001). The three groups were

fed for 105 days as follows:

1. Control ’C’ group animals were fed a ration having concentrate mixture

(Maize 28, Bajra 5, GNC 10, Mustard cake 13, Soybean meal 15, Wheat bran

15, Rice bran/ polish 11, Mineral mixture 2 and Common salt 1part), and

wheat straw in the ratio of 50:50, along with 10 kg green fodder/animal/day to

meet the vitamin A requirements as per NRC (2001).

2. Experimental Group ‘CSC’ T-1 was fed a ration as in above, but 10 parts of

soya meal of concentrate mixture of control was replaced by expeller

cottonseed cake.

3. Experimental Group ‘DCSC’ T-2 was also fed with a similar ration as in

control except that in this soya meal of concentrate mixture was partially

replaced by 10 parts of decorticated cottonseed cake (DCSE) solvent

extracted.

The experimental feeding started after 15 days of acclimatization.

3.4.3 Housing of animals

All the animals were housed in the experimental sheds of NDRI, Karnal

for 105 days which was well ventilated having individual pens to facilitate

individual feeding. Proper cleanliness and healthy surroundings were ensured

throughout the experimental period. Body weights were recorded at fortnightly

intervals in the morning hours before offering any feed or water.

An initial adaptation period of about 90 days was given to the animals

in shed. Then animals were shifted to metabolic cages for seven days

metabolism trial in between the experimental period.

Photo : Decorticated Cottonseed Cake Solvent Extracted

28

Table 3.4.4: Description of experimental animals

Group Animal No. Date of birth Age

(months)

Initial B.wt.

(kg)

Control

7562 19/04/12 7.5 89

7575 03/07/12 5 70

7517 21/09/11 15 178

7560 04/04/12 8 122

7516 19/09/11 15 167

7582 28/08/12 4 60.2

Avg. B.wt. (kg) 9.08±1.97

114.33±20.34

T 1

7546 26/02/12 10 130

7580 09/08/12 4 75

7581 21/08/12 4 60

7520 05/10/11 14 125

7519 02/10/11 14 213

7574 29/06/12 6 76

Avg. B.wt. (kg) 8.66±1.9

113.16±23.13

T2

7563 20/04/12 8 80

7577 14/07/12 5.5 65

7523 20/11/11 13 153.3

7578 08/08/12 4.5 93

7576 11/07/12 5.5 68

7514 02/09/11 15 233

29

3.5 Metabolism trial

Animals were shifted to metabolism cages after 12 weeks of preliminary feeding

for the metabolism trial for seven days. Animals were weighed before and after the trial.

Respective TMRs were offered to each animal as explained earlier. Fresh drinking

water was provided twice a day and the quantity was measured each time to calculate

the total water intake.

3.5.1 Sampling, processing and storage of feed samples

The samples of TMR offered, residues, if any, were taken daily for DM estimation

during metabolism trial. These samples were pooled at the end of the collection period

and ground to pass through 1 mm sieve and stored in air tight containers. The samples

were analyzed for proximate principles (OM, CP, Ash, CF and EE) and cell wall

constituents (NDF and ADF) as per standard procedures to calculate the %TDN.

Faeces voided during 24 h was collected daily for seven days and weighed at

8:30 am daily. After thorough mixing, an approximately 2% of total sample on weight

basis was kept for DM estimation. Dried pooled faecal samples were ground to pass

through 1 mm sieve size and analyzed for proximate principles and cell wall

constituents as per standard procedures. For N estimation of faeces, measured

quantities of faecal samples were collected daily for seven days and stored in plastic

containers containing 25 ml of 25% H2SO4 solution.

Total urine voided from each animal was collected using urine collection cans.

Total urine output for 24 h was measured daily at 9:00 am and an aliquot (1% of total

output) was taken for the nitrogen estimation. The aliquots were stored in plastic

containers containing 10 ml of 25% H2SO4 for total nitrogen estimation.

3.5.2 Analysis of feed, faeces and urine

Dry matter, organic matter, crude protein, ether extract, and total ash in feeds

and faeces samples were determined (AOAC, 2005). The fractions of cell wall

constituents (NDF, ADF and ADL) were also determined (Van Soest et al., 1991). Urine

samples were analyzed for nitrogen content (AOAC, 2005).

Photo : Animal during Metabolic Trial

30

3.5.3 Estimation of total nitrogen in urine

From each sample 5 ml urine was transferred in to Kjeldahl digestion

tube, digested by adding 20 ml concentrated H2SO4 along with 2- 3 g

digestion mixture. Digested material was diluted with distilled water up to 100

ml in volumetric flask. An aliquot (10 ml) of digested sample was taken in

distillation tube and adequate amount of 40% NaOH was added into the tube

and distillation was done. The distillate was collected in a conical flask

containing 2% boric acid solution having mixed indicator, which was titrated

against N/100 H2SO4. Total-N was calculated as below:

V X B X DF X 0.014 X 100 X 1000

Total-N (mg/100 ml) =

Vol. of sample taken for digestion

V = vol. of H2SO4 used

B = normality of H2SO4

DF= dilution factor (total volume made/ aliquot taken for distillation)

3.5.4 Calculation of nutrient digestibility (%)

The nutrient (DM, OM, CP, CF, EE, NFE, NDF and ADF) digestibility

coefficients were calculated from the nutrient intake and nutrient outgo in

faeces during metabolism trial as below

(Nutrient intake – Nutrient outgo in faeces) X 100

Digestibility (%) =

Nutrient intake

3.5.5 Estimation of total digestible nutrient (TDN %)

TDN (%) = DCP + DCF + DNFE + (DEE × 2.25)

3.5.6 Periodical studies

Daily dry matter intake and fortnightly body weights were recorded.

Feed conversion ratio (FCR) in Karan fries calves was calculated as kg of

feed consumed (DM basis) per kg of live weight gain for 105 days study.

31

3.6 Haematological Parameters

During 105 days of feeding, the blood samples were collected at an

interval of 30 days from all the animals by jugular puncture in heparinised

vaccutainer, mixed well and brought to the laboratory after placing on ice. Just

after blood collection, the tubes were rotated between palm to ensure proper

mixing of blood and anticoagulant. The samples were kept in the ice- box after

collection. Haematological parameters were analyzed immediately in fresh

blood sample in automatic blood analyzer. Then, the samples were

centrifuged at 3000 rpm for 15 minutes to separate the plasma. The plasma

samples were stored at -20°C for the estimation of glucose, blood urea

nitrogen (BUN), total protein, albumin, and globulin.

Whole blood was analyzed using BC-2800 Vet Auto Haematological Analyzer.

Chemicals used for analyzer were purchased from M/s RFCL Limited, New

Delhi. The analysis was made within one hour of blood collection. The

haematological parameters determined by analyzer were white haemoglobin

(HGB).

3.6.1 Albumin

Albumin was estimated in blood plasma samples using kit supplied by

Span Diagnosis Ltd. Albumin binds with anionic dye Bromocresol Green

(BCG) to form green color complex which is measured at 630nm.

Albumin + BCG →green colored complex

Kit reagents were prepared and stored as per the instructions provided

with the assay kit.

Procedure

Pipette into tube

marked

Blank Standard Test

Serum / Plasma - - 10 µl

Albumin standard - 10 µl -

Albumin reagent 1000 µl 1000 µl 1000 µl

The content were mixed well and incubated at room temperature (15-

300C) or 1 min.

32

UV- spectrophotometer was blanked with reagent blank and the

absorbance of standard and test sample was measured.

Calculation

The concentration of albumin was calculated as per formula and

expressed in mg/dl

Plasma albumin (g/dl) = X 4

Concentration of standard=4 g/dl

3.6.2Total protein

Total protein was estimated in blood plasma samples using kit supplied by

Span Diagnosis Ltd. The peptide bonds of proteins react with cupric ion in

alkaline solution to form a colored chelate, which is measured at 578 nm.

Protein + Cu²+ → Cu-protein complex


with the assay kit.

Procedure

Pipette into tube

marked

Blank Standard Test

Serum / Plasma - - 10 µl

Protein standard - 10 µl -

Biuret reagent 1000 µl 1000 µl 1000 µl

The contents were mixed well and incubated at 370C temperature for 5

min.



Calculation

The concentration of total protein was calculated as per formula and

expressed in g/dl

Absorbance of test

Absorbance of standard

33

Total protein concentration (g/dl) = X 6.5

Concentration of standard = 6.5 g/dl

3.6.3 Globulins = Total protein – albumin

3.6.4 Estimation of blood glucose

Plasma glucose was estimated using GOD-POD kit (Span Diagnostics

Ltd., India). In this procedure, glucose oxidase (GOD) oxidizes glucose to

gluconic acid and hydrogen peroxide. In presence of enzyme peroxidase,

released hydrogen peroxide is coupled with phenol and 4-amino antipyrine (4-

AAP) to form colored Quinoneimine dye. Absorbance of colored dye

measured at 505 nm is directly proportional to glucose concentration in the

sample.

Procedure

10 µl of plasma aliquots were pipetted in 10x75 mm tubes (in

duplicate), to which1000 µl of working glucose reagent was added. The

contents were mixed well. The tubes were then incubated at room

temperature (15-30°c) for 30 minutes. Along with the unknown sample, blank

and standard tubes (in duplicate) containing 10µl of double distilled water and

10µl of standard (100 mg/dL) respectively were processed identically. UV-

Spectrophotometer (Bio-Age 756PC) was blanked with reagent blank and the

absorbance of standard and unknown sample was measured.

Concentration was calculated as per formula and expressed as mg/dl

Absorbance of test

Plasma glucose (mg/dL) = χ 100


3.6.5 Estimation of blood urea nitrogen (BUN)

BUN was estimated in plasma samples by GOD-POD kits obtained from

Span Diagnostics Ltd. Urea hydrolyses to ammonia in presence of urease

Absorbance of test


34

enzyme which reacts with hypochlorite and phenolic chromogen in alkaline

medium to form colored complex which is measured at 578 nm.


with the assay kit. The procedure is presented below:

10 µl of plasma aliquots were pipette in 10×75 mm tubes in duplicate,

to which 1000 µl of working BUN solution-1 was added.

Blank (10 µl distilled water) and standard (10 µl from standard

50mg/dL) were pipetted in duplicate, to which 1000 µl of working BUN

solution-1 was added.

The contents were mixed well and incubated at 37°C for 3 min.

1000 µl of working BUN solution-2 was added to each tube.

The contents were mixed well and incubated at 37°C for 5 min.



Plasma BUN levels were calculated as per formula and expressed in

mg

Plasma BUN (mg/dl) = X 50

Concentration of standard= 50 mg/dl

3.7 STATISTICAL ANALYSIS

The results obtained during this study were analyzed by analysis of

variance (ANOVA) using software package SPSS version 16.0, 2010.

Absorbance of test


CHAPTER CHAPTER CHAPTER CHAPTER –––– 4 4 4 4

RESULTRESULTRESULTRESULTSSSS AND DISCUSSION AND DISCUSSION AND DISCUSSION AND DISCUSSION

35 Results & Discussion

RESULTS AND DISCUSSION

4.1 PERCENT INGREDIENT AND ANALYSIS OF CONCENTRATE

MIXTURES

A concentrate mixture based on maize as the major energy ingredient

and soybean meal (de-oiled) as protein supplement was formulated. The

Control ‘C’ concentrate mixture consisted of (kg/ton) maize 280, bajra 50,

mustard oil cake130, groundnut cake 100, wheat bran 150, rice polish 110,

soybean meal 150, mineral mixture 20 and common salt 10. It was not having

any feed additive. The 100 parts (kg/ton) of soybean meal was replaced with

cottonseed cake expeller (CSC) for experimental group ‘T-1’ or with

decorticated cottonseed cake solvent extracted (DCSC) for experimental

group ‘T-2’ (Table 4.1) to study the effects of partial replacement of soybean

meal (SBM) with these cakes available as protein supplements at a lower

price than soybean meal (Table 4.1). Additional Urea was added in group ‘T-

1’ and ‘T-2’ to make the concentrate mixtures isonitrogenous.

The percent chemical composition and cell wall contents of different

ingredients used in concentrate mixtures is depicted in Table 4.2 and percent

chemical composition of Control ‘C’, ‘T-1’ and ‘T-2’ concentrate mixtures and

also of green oats and wheat straw fed to crossbred cows is given in table

4.3. The main roughage was wheat straw. However, chopped (2-3cm size)

green oats, a non-leguminous fodder was also a source of roughage to meet

the vitamin A requirements of the crossbred cows. Weighed quantity of wheat

straw mixed with each separate concentrate mixture (‘C’, ‘T-1’ or ‘T-2’) in the

ratio of 50:50 was offered individually to three groups of crossbred calves and

10kg chopped oats fodder per animal per day was also mixed to make

complete feed mixtures for all the groups of crossbred calves.

The diets pertaining to the concentrate mixtures of three experimental

groups; green oats and wheat straw showing chemical composition and cell

wall contents was Dry matter 90.2, 90.3, 91.1,16.0 and 90.6, organic matter

90.4, 90.9, 90.2, 88.5 and 91.6, crude protein 19.8, 20.0, 19.9, 8.3 and 2.6,

ether extract 4.2, 6.2, 4.8, 2.7 and 0.87, crude fibre 10.8, 10.4, 9.4, 42.3 and

33.1, Neutral detergent fibre 36.2, 31.7, 33.8, 63.2 and 81.6, acid detergent


fibre 15.1, 15.2, 12.7, 43.0 and 57.1, hemicellulose 21.1, 16.5, 21.1, 20.2 and

24.5 percent in control ‘C’, ‘T-1’, ‘T-2’, oats fodder and wheat straw

respectively.

Table 4.1:- Percent ingredient composition of experimental concentrate

mixtures fed to female crossbred calves

Ingredients

Groups

Price (Rs/qnt) SBM

‘C’

CSC

‘T-1’

DCSC

‘T-2’

Maize grain 28 28 28 1480

Bajra 5 5 5 1500

Mustard oil cake 13 13 13 1800

Ground nut cake 10 10 10 3250

Wheat bran 15 15 15 1380

Rice polish 11 11 11 1300

Soybean meal 15 5 5 3500

Cotton seed cake

(Expeller) - 10 - 1900

Decorticated cottonseed

solvent extracted - - 10 2000

Salt 1 1 1 300

Mineral mixture 2 2 2 5300

Urea - 1 0.25 700

The Table 4.3 clearly depicts that the three concentrate mixtures i.e. ‘C’, ‘T-1’

and ‘T-2’ having soybean meal or soybean meal partially replaced with

cottonseed cake (expeller) or decorticated cottonseed cake (solvent

extracted) respectively had medium level of crude protein (19 to 20 percent),

ether extract (4.2 to 6.2%), nitrogen free extract (52.0 to 56.3) showing not

much variation in chemical compositions.


Table 4.2 Percent chemical composition and cell wall contents of

different ingredients used in concentrate mixture (on DM basis)

Ingredients

DM

OM

EE

CP

TA

AIA

NDF

ADF

Hemi -

cellulose

Maize 91.76 97.20 4.45 10.37 2.20 0.28 18.25 9.34 8.91

Bajra 90.58 97.67 3.85 11.80 2.43 0.43 37.42 5.35 32.07

Ground nut cake (Expeller)

91.62 92.74 7.36 41.50 6.07 1.52 22.42 14.32 8.10

Cotton seed cake (Expeller)

90.32 94.50 7.33 25.55 5.50 0.38 38.70 37.52 1.18

Decorticated cottonseed cake

(Solv. Ext.) 90.17 92.90 0.70 44.05 7.10 0.81 31.00 18.00 13.00

Mustard oil cake 91.51 93.86 10.44 37.30 6.24 1.04 23.32 21.62 1.70

Soybean meal (Deoiled)

90.26 94.09 1.79 45.11 5.01 0.53 17.49 16.62 0.87

Wheat bran 90.52 93.90 2.55 17.10 6.10 0.51 33.86 16.34 17.52

Rice polish 88.07 91.89 1.13 17.50 9.21 1.26 35.27 18.45 16.82

Thus all these concentrate supplied higher level of nitrogen as well as readily

available energy to the growing cross bred female calves for rapid growth.

Wheat straw being highly fibrous (NDF 81.6%, ADF 57.1% and crude fibre

33.1%) may release less energy slowly. The hemicellulose content of the

wheat straw 24.5% compared to the concentrate mixture (16.1 to 21.1)

indicating that wheat straw was also a potential source of digestible energy,

but due to high contents of acid detergent lignin (Sehgal and Punj,1983, Dey


2002) the hetero polysaccharide was not available in plenty to the ruminants.

The values for crude protein and total ash (TA) contents of groundnut cake

and mustard oil cake in the present study were quite comparable (Dutta and

Singh, 1994; Dey, 2002).

Table 4.3 Percent chemical composition of concentrate mixtures SBM

‘C’, ‘T-1’ (replacing 10 parts of soybean meal with cottonseed cake

expeller), ‘T-2’ (replacing 10 parts of soybean meal with decorticated

cottonseed cake), green oats and wheat straw (on DM basis)

Parameters

Groups Oat

fodder

Wheat

straw SBM

‘C’

CSC

‘T-1’

DCSC

‘T-2’

Dry matter 90.2 90.3 91.1 16.0 90.6

Organic matter 90.4 90.9 90.2 88.5 91.6

Crude Protein 19.8 20.0 19.9 8.5 2.6

Crude fibre 10.8 10.4 9.4 42.3 33.1

Ether extract 4.2 6.2 4.8 2.7 0.9

Nitrogen free

extract 56.3 52.0 55.8 44.2 45.8

Total ash 9.6 9.1 9.8 11.5 8.4

Acid insoluble ash 1.8 1.4 1.1 3.6 4.0

Neutral detergent

fibre 36.2 31.7 33.8 63.2 81.6

Acid detergent

fibre 15.1 15.2 12.7 43.0 57.1

Hemicellulose 21.1 16.5 21.1 20.2 24.5


The oats green fodder had only 6.38 percent lignin (Dey, 2002) due to

which it may supply sufficient amount of available energy from hemicellulose

(20.2%) and crude fibre (42.3%). As the oat was fodder little matured it

contained only 8.5 percent crude protein in the present study. The percent

chemical composition of green oats fodder observed in this study was similar

to as reported by earlier research worker (Tripathi et al., 2000, Sachin, 2012).

The proximate principles of wheat straw (Table 4.3) viz crude protein

(2.6%), crude fibre (33.1%), NFE (44.2%) and total ash (11.5%) were in close

agreement with the value reported in the literature i.e. 2.5 to 4.5, 30.5 to 48.9,

37.5 to 57.4 and 7.2 to 10.9 percent respectively (Sehgal and Makkar,1994,

Sehgal et al., 1999, Dey, 2002, Tripathi et al., 2007). The cottonseed cake

(expeller) used was having 25.55% crude protein and 7.7% ether extract and

thus was observed to be a quality protein supplement. It has also got higher

undegradable protein compared to groundnut cake and mustard oil cake

(Sehgal and Makkar, 1994). The crude protein content of soybean meal de-

oiled was 45.11% quite comparable to the decorticated cottonseed cake

solvent extracted (44.05%), though a difference in the purchase price (Table

4.1) was quite high (Rs 38/- vs Rs 20/- per kg).

4.2 IN VITRO STUDIES

The in vitro studies were conducted to measure the rumen dry matter,

organic matter and crude protein degradability of protein supplements viz

Soybean meal (de-oiled), cottonseed cake (expeller), decorticated cottonseed

cake solvent extracted and of three concentrate mixture prepared using these

supplements being fed to female cross bred calves in groups as ‘C’, ‘T-1’ and

‘T-2’ respectively. The 1st stage of Tilley and Terry (1963) was used and the

protein supplements and concentrate mixtures were incubated for 48h. (Table

4.4/ 4.4a). The data showed that the soybean meal and concentrate mixture

‘C’ had higher (P≤0.05) DM (64.41±1.61, 71.41±0.26), OM (64.49±0.58,

67.03±0.85) and CP(64.46±0.60, 65.74 ±1.73) degradability followed by

cottonseed cake (expeller) and conc. mixture (T-1) i.e. DM (59.58±1.02,

67.92±1.10), OM (52.86±1.58, 58.28±0.94) and CP (58.96±1.98, 59.04±1.41)

and decorticated cottonseed cake solvent extracted and concentrate mixture


‘T-2’ had lowest (P≤0.05) DM (55.70±0.45, 65.05±0.93), OM (53.31±0.70,

55.03±0.91) and CP (55.92±0.87, 56.54±1.25) degradability. The data clearly

showed that decorticated cottonseed cake solvent extracted protein

supplement had higher rumen undegradable protein. Cottonseed cake

(expeller) also had more undegradable protein than soybean meal de-oiled.

Thus decorticated cottonseed cake solvent extracted can be fed to the

growing animals requiring rapid rate of growth with advantage compared to

soybean meal de-oiled as the more crude protein will be available at the site

of absorption. Torane et al. (2006) reported that RDP: UDP ratio of 52:48 in

undecorticated cottonseed cake stimulated higher growth rate in crossbred

calves because of higher rumen undegradable protein availability from this

cake. However, in the present study the cottonseed cake expeller had RDP:

UDP ratio of 59:41 compared to 65:35 in soybean meal and 56:44 in

decorticated cottonseed cake solvent extracted (Table 4.4). Sehgal and

Makkar (1994) reported a RDP: UDP ratio of 61:39 in cottonseed cake

(expeller) which was quite comparable to the RDP: UDP ratio of 59:41 in the

present study.

Table 4.4 In vitro degradability of different cakes after incubation at 48

hrs.

In vitro degradability

Ingredients Dry matter Organic matter Crude protein

Soybean meal 64.41c±1.61 64.49a±0.58 64.46b±0.60

Cottonseed cake

(Expeller) 59.58b±1.02 52.86a±1.58 58.96a±1.98

Decorticated

Cottonseed cake

(Solv. ext.)

55.70a±0.45 53.31a±0.70 55.92a±0.87

SEm 1.38* 1.97* 1.41*

Values bearing different superscripts, a, b, c in a column differ significantly (P < 0.05) *Significant (P < 0.05)


4.4 (a) In vitro degradability of different Concentrate mixtures after incubation at 48 hrs.

In vitro degradability

Conc. Mixtures Dry matter Organic matter Crude protein

SBM ‘C’ 71.41b±1.26 67.03c±0.85 65.74b±1.73

CSC ‘T-1’ 67.92a±1.10 58.28b±0.94 59.04a±1.41

DCSC ‘T-2’ 65.05a±0.93 55.03a±0.91 56.54a±1.25

SEm 1.07* 1.84* 1.56*

Values bearing different superscripts a, b, c in a column differ Significantly (P < 0.05) *Significant (P < 0.05) SBM: Soybean Meal CSC: Cotton Seed Cake (Expeller) DCSC: Decorticated Cotton Seed Cake (Solv. ext.)

4.3 GROWTH STUDIES

Average fortnightly body weights of female cross bred (Tharparkar X

Holstein Friesion) calves fed concentrate mixture ‘C’, ‘T-1’ and ‘T-2’ with

wheat straw and green oats fodder as complete feed mixture individually in

three groups ‘C’, ‘T-1’ and ‘T-2’ have been shown diets in table 4.5.

Fortnightly changes in body weights of these calves of group ‘C’, ‘T-1’ and

‘T-2’ diet have been illustrated graphically in figure 1.

At the start of the experiment, the average body weight were 114.37 +

20.35,113.17+ 23.13 and 115.37+ 26.96 for ‘C’, ‘T-1’ and ‘T-2’ diet groups

respectively. The average live weight of the female cross bred calves at the

end of 105 days of experimental period were recorded to be 181.00 + 23.63,

174.50+ 28.13, and 194.13+ 34.19kg in ‘C’, ‘T-1’ and ‘T-2’ diet groups

respectively. These cross bred calves of ‘C’ and ‘T-2’ diets showed higher

gain in body weight (66.6+ 3.96 and 78.8+ 10.32kg) then ‘T-1’ (61.3+ 6.5kg)

diet indicating significantly (P< 0.05) higher growth in ‘T-2’ diet then ‘C’ and ‘T-

1’ diet group respectively (Table 4.6).


Table 4.5 Effect of partial replacement of soybean meal (SBM) with

cottonseed cake (CSC) or decorticated cottonseed cake (DCSC) on

fortnightly body weight changes (kg) in female cross bred calves

Groups

Fortnights SBM ‘C’ CSC ‘T-1’ DCSC ‘T-2’

0 114.37±20.35 113.17±23.13 115.37±26.96

1st 125.13±21.58 124.33±24.21 129.87±28.40

2nd 134.97±21.89 134.17±25.44 139.83±30.07

3rd 146.50±22.80 142.37±26.07 151.82±30.84

4th 151.80±22.70 153.32±26.57 159.83±31.08

5th 162.42±24.28 161.40±27.70 172.03±30.49

6th 173.50±23.40 166.83±28.54 181.83±32.58

7th 181.00±23.63 174.50±28.17 194.17±34.19

Also the average daily gains were observed to be higher (P< 0.05) in

‘T-2’ diet (750+51g) compared to ‘C’ (634+61g) and ‘T-1’ (583+37g) diets and

the differences were statistically significant (P< 0.05). Thus there was an

improvement of 18.3 percent and 28.6 percent in body weight gains of cross

bred calves fed decorticated cottonseed cake ‘T-2’ diet over that SBM of ‘C’

and CSC ‘T-1’ diet. Growth performance (fig. 1) revealed that the initial

differences in body weight gain in ‘C’ and ‘T-1’ diet were narrow, but ‘T-2’ diet

since the start of the experiment at each fortnight of the 105 days

experimental period showed higher body weight gains compared to other two

dietary groups ‘C’ and ‘T-1’ and even after 3rd fortnight the growth

performance of ‘T-1’ diet was lower than ‘C’ diet group indicating that partial

replacement of soybean meal with cottonseed cake (Expeller) was not

advantages to growing female calves. This may be attributed to 1% urea

addition for making the dietary treatment isonitrogenous and the farm bred

Figure 1 Effect of partial replacement of soybean meal (SBM)with cottonseed

cake (CSC) or decorticated cottonseed cake (DCSC) on fortnightly body weight

changes in female cross bred calves

110

120

130

140

150

160

170

180

190

200

0 1 2 3 4 5 6 7

Bo

dy

wt.

(kg

)

Fortnights

SBM

CSC

DCSC

43

animals may not have relished the sudden given urea diet. The total

dry matter intake was also low in this dietary treatment (‘T-1’ diet).

Growth is influenced by so many factors such as nutritional, hormonal,

managemental, biochemical, genetical and environmental. The other factors

remaining the same, it is only nutrition which might affect the growth rate. The

general growth curve may be divided in to self accelerating phase of

increasing slope and a self inhibiting phase of decreasing slope. Growth curve

is sigmoid in nature. The present study pertains only to be the accelerating

phase of growth (fig.1). Thus the difference in growth performance in three

groups of animals offered ‘C’ ‘T-1’ and ‘T-2’ diets respectively could be

attributed to the better utilization of partial replaced soybean meal with 10

parts of decorticated cottonseed cake solvent extracted in concentrate

mixture ‘T-2’ in a complete feed mixture than cottonseed cake expeller ‘T-1’.

The difference in growth performance was attributed to the feed intake

also. The animals of group ‘T-1’ diet eat less dry matter (4.50+ 0.58) than ‘T-2’

indicating higher feed efficiency of ‘T-2’ diet. Contrary to this Gonzalez et al.

(1988) showed that small quantity of cottonseed cake efficiently improved the

utilization of low quality forage and the performance of animals, but in the

present study the % feed efficiency was found to be higher with 10 parts

replacement of soybean meal with decorticated cottonseed cake solvent

extracted and not with cottonseed cake expeller as compared to soybean

meal containing dietary treatment ‘C’.

4.4 EFFECT OF PARTIAL REPLACEMENT OF SOYBEAN MEAL WITH

COTTONSEED CAKE OR DECORTICATED COTTONSEED CAKE ON

VOLUNTRY FEED INTAKE, PRODUCTIVE PERFORMANCE AND

PERCENT FEED EFFICIENCY

The total dry matter intake, animal productivity in terms of growth of

cross bred calves; percent feed efficiency (measured as the gain in weight

(kg) per 100kg of feed intake) or feed conversion ratio (measured as the

DMI/kg gain) of cross bred calves kept on three different feed regimes viz

control ‘C’, ‘T-1’, and ‘T-2’ diet for 105 days experimental period has been

shown in table 4.6.

44

The calves in C’, ‘T-1’, and ‘T-2’ diet were offered a weighed quantity of

wheat straw and concentrate mixture in the ratio of 50:50 and 10kg green oats

per calf per day and actual total dry matter consumption for the 105 days

experimental period (434.9+42.00, 402.8+ 43.30, 425.1+ 41.40kg) and dry

matter intake per day per cross bred calves (4.14+ 0.40, 3.83+ 0.44, 4.04+

0.39kg) remained statistically same in all the groups, the calf performance in

terms of growth rate ‘g’ (634+ 61, 583+ 37, 750+ 51) in C’, ‘T-1’, and ‘T-2’

differed (P< 0.05) and it was better in the ‘T-2’ group in which 10 parts of

soybean meal was partially replaced by decorticated cottonseed cake ( solv.

ext.) over control and ‘T-1’ diet (Table 4.6). The productive performance in

terms of growth of crossbred calve in CSC ‘T-1’ diet was not better than the

other two dietary group ‘C’ and ‘T-2’ numerically little less dry matter intake

per day/calf was observed in ‘T-1’ diet which may have reflected in the lower

growth rate. Wanapat et al. (1996) reported that straw intake was significantly

(P< 0.05) decreased from 1.25% to 0.88% of live weight as the level of

cottonseed cake was increased from 2 to 5kg /day in dairy cows. However a

summary of 18 feeding trials (Coppock et al., 1987) with whole cottonseed

showed no significant reduction (P> 0.05) in dry matter intake when whole

cottonseed was included up to 25% of the ration, But Shirley et al.(1998)

reported no difference in dry matter intake when feeding expended expelled

cottonseed cake in place of whole cottonseed. Similarly other studies (Saij

Paul et al, 2006; Sullivan et al., 1993) did not find any difference in DMI when

feeding whole linted cottonseed and cracked or ground Pima cottonseed. As

in the present study Arajuo et al. (2004) also observed no influence of the

processing of cottonseed on the nutrient intake.

45

Table 4.6 Productive performance of female crossbred calves fed with

cottonseed cake (CSC) or decorticated cottonseed cake (DCSC) as a

partial replacement of soybean meal SBM) in complete feed mixtures.

Particular

Groups

SEm

(P < 0.05)

SBM

‘C’

CSC

‘T-1’

DCSC

‘T-2’

No. of animals 6 6 6

Avg. initial body weight (kg) 114.3± 20.30 113.2± 23.10 115.36± 26.90 12.82 NS

Avg. final body weight (kg) 181±23.60 174.5± 28.10 194.1± 34.20 15.85 NS

Total Wt. gain(kg) 66.6ab± 3.96 61.3a± 6.50 78.8b± 10.32 4.39*

Average daily gain ‘g’ 634 ab± 61 583a± 37 750b± 51 41.81*

Dry matter intake(DMI) (kg)

a) From concentrate** 181.6±19.20 164.38±22.60 172.8±20.20 11.37 NS

b) From wheat straw** 135.4±20.53 121.4±22.20 130.2±20.70 11.57 NS

c) From green oats** 117.8±3.10 117±4.80 122.1±1.50 1.93 NS

Total dry matter intake** 434.9±42.00 402.8±46.30 425.1±41.40 23.72 NS

Average dry matter intake/day 4.14±0.40 3.83±0.44 4.04±0.39 0.22 NS

Average dry matter

intake/100kgBW 2.50±0.42 2.36±0.45 2.71±0.57 0.26 NS

Average dry matter intake/kg

W0.75(g) 89.10±12.74 83.41±14.46 93.88±17.34 71.53 NS

Feed conversion ratio (kg

DMI/kg gain) 6.26±0.52 6.75±1.08 6.69±1.07 0.51 NS

% Feed efficiency (kg gain/

100kg feed intake ) 16.21ab±2.13 15.34a±0.69 18.55b±1.73 0.95*

Means with different superscripts a, b, ab in a row differ significantly (P < 0.05) NS- Non significant at 5% level **105 days

Contrary to this Chaudhary et al. (2009) found significant higher dry

matter intake in lactating Surti buffaloes which were fed 60% cottonseed cake

46

in concentrate mixture. Zhang and Co-workers (2007) observed that dry

matter intake was not altered when cottonseed was included up to 25% of the

diet. The percent feed efficiency (kg gain/100kg feed intake) was found to be

higher (18.55+1.75) in the group ‘T-2’ in which decorticated cottonseed cake

solvent extract replaced 10 parts of soybean meal. However, statistically no

difference was observed between ‘C’ and CSC ‘T-1’ diets. Though did not

significant average DMI /100kg BW was also found to be higher in ‘T-2’ diet

compared to other indicating higher palatability of decorticated cottonseed

cake in crossbred calves.

4.5 EFFECT ON NUTRIENT INTAKE AND NUTRIENT UTILIZATION

Effect of partial replacement of soybean meal ‘C’ with cottonseed cake

‘T-1’ or decorticated cottonseed solv. ext. ‘T-2’ on nutrient intake /day (kg) in

female crossbred calves during metabolic trial period has been depicted in

Table 4.7. The data showed that the nutrient intake and DMI /100kg body

weight did not differ significantly in all the three groups ‘C’, ‘T-1’, and ‘T-2’

and thus accordingly except Ether extract, no other nutrient intake differed.

Though numerically TDN intake was found to be then more in ‘C’ then others;

the difference was not significant.

Crude fat (ether extract) intake differed significantly (P< 0.05) and it

was higher in ‘C’, ‘T-1’, and ‘T-2’ diet (0.28±0.03, and 0.27±0.03kg) than

control ‘C’ (0.14±0.01 kg) diet.

The variation present in the digestibility coefficients of dry matter,

organic matter, crude protein, crude fibre and nitrogen free extract were within

the range, and no significant differences were observed in the digestibility of

these nutrients among the various groups of animals. Dry matter digestibility

(%) (63.63±1.32, 61.19±1.10 and 60.62±1.63 ), organic matter

digestibility (%) (69.35 ±1.38, 67.42±1.40 and 66.97±1.47), crude protein

digestibility (%) (59.77±2.51, 59.7±1.93 and 61.00±2.73), crude fibre

digestibility(%) (64.51±1.70, 58.87±1.25 and 61.69 ± 2.44) and NFE

digestibility (%) (70.24 ± 1.20, 65.84 ± 1.50 and 70.13 ± 1.90) in control ‘C’,

‘T-1’ and ‘T-2’, respectively. However, digestibility of ether extract was

significantly higher (87.16±1.04 and 86.39±0.93) in ‘T-1’ and ‘T-2’ than ‘C’

(79.34±1.43). Similar to the digestibility coefficient of nutrient viz DM, OM, CP,

47

CF, except EE. The digestibility coefficient cell wall content i.e. NDF, ADF of

‘C’ ,’T-1’ and ‘T-2’ diets, also did not differ significantly but showed numerically

higher value for diet ‘C’ compared to ‘T-1’ and ‘T-2’ (Table 4.8). There was no

difference in the digestibility of ADF between ‘T-1’ and ‘T-2’ in which the

cottonseed cake expeller or decorticated cottonseed cake solvent extracted

was added as a 10 part replacement of soybean meal. Thus all the three diets

were found to be equally good as for as the nutrient digestibility was

concerned.



nutrient intake (kg) in female cross bred calves (during metabolic trial)

Parameters Groups SEm

(P <0.05) Nutrient

intake(kg) SBM ‘C’

CSC ‘T-1’

DCSC ‘T-2’

Dry matter 4.80±0.33 4.38±0.45 4.50±0.58 0.26NS

Crude protein 0.54±0.03 0.53±0.05 0.50±0.06 0.03NS

Ether extract 0.14±0.01a 0.28±0.03b 0.27±0.03b 0.02*

Crude fibre 1.32±0.09 1.21±0.13 1.22±0.17 0.07NS

Nitrogen Free

Extract 2.71±0.16 2.29±0.20 2.46±0.23 0.12NS

Total Digestible

Nutrient 3.20±0.16 2.66±0.26 2.62±0.23 0.14NS

Means with different superscriptsa,b in a row differ significantly (P <0.05) NS- Non significant at 5% level *Significant at 5% level

Kakkar and Mudgal, (1997) and Mudgal and Mulla (1985) observed no

significant (P< 0.05) difference in the intake or digestibility coefficients for dry

matter, organic matter, crude protein, crude fibre and nitrogen free extract.

However, they observed a significant difference in the utilization of ether

extract. Kehar et al. (1950-51) reported that crude fibre digestibility of

48

decorticated cottonseed cake and undecorticated cottonseed was similar. In

contrast Kakkar and Mudgal, (1997) observed higher crude fibre digestibility

of decorticated cottonseed cake than undecorticated cottonseed cake as well

as whole cotton seed.

Similarly, Suliman and Babiker (2007) also did not observe difference

which range between 1.11 to 1.18 kg/day in fattening lambs fed with different

protein supplements like groundnut cake, sesame cake, cottonseed cake and

sunflower seed cake. Ward et al. (2008) reported no change in dry matter

intake by Barki lambs fed diet containing soybean meal, cottonseed meal and

cottonseed meal supplemented with ferrous sulphate though slightly lower dry

matter intake was observed in lambs fed soybean diets.

Digestibility of crude fat (ether extract) differed significantly (P<0.05)

and was higher in ‘T-1’ and ‘T-2’ (79.34±1.43 and 87.16±1.04) than control ‘C’

diet (86.39±0.93%). Digestibility of dry matter, organic matter, crude protein

and nitrogen free extract did not differ significantly (P<0.05). similar to the

findings in the present study Bangani et al. (2000) found no difference in

respect of dry matter intake and feed conversion efficiency in groups fed with

cottonseed cake and soybean oil cake meal to the Holstein and jersey heifer

calves. El-din et al. (1992) and Binondi et al. (1993) also reported no

difference between diets containing cottonseed oil cake meal and soybean oil

cake meal in Holstein bull calves.

Philips and Rao (2001) also observed no difference in dry matter

digestibility in lambs fed with diets containing pigeon pea, cottonseed meal or

alfalfa as protein sources. Ward et al. (2008) reported that un-decorticated

CSM supplemented with ferrous sulphate significantly improved the nutrient

digestibility in growing Barki male lambs compared to those lambs fed un-

decorticated CSM or SBM without the addition of ferrous sulphate.

Numerically lower intake and lower growth rate in cottonseed cake (expeller)

diet ‘T-1’ than ‘C’ and diet ‘T-2’ can also be due to no addition of ferrous

sulphate in diet ‘T-1’ which may have higher free gossypol.

49



digestibility coefficient (%) of various nutrients of different complete

feed mixture in female cross bred calves.

Particulars

Groups SEm

(P <0.05) SBM ‘C’

CSC

‘T-1’

DCSC

‘T-2’

Dry matter 63.63±1.32 61.19±1.10 60.62±1.63 0.81 NS

Organic matter 69.35±1.38 67.42±1.40 66.97±1.47 0.80 NS

Crude protein 59.77±2.51 59.7±1.93 61.00±2.73 1.32 NS

Ether extract 79.34±1.43a 87.16±1.04b 86.39±0.93b 1.05*

Crude fibre 64.51±1.70 58.87±1.25 61.69±2.44 1.16 NS

Nitrogen Free

Extract 70.24±1.20 65.84±1.50 70.13±1.90 0.98 NS

Acid detergent fibre 50.93±1.86 48.96±1.49 46.08±2.71 1.22 NS

Neutral detergent fibre

60.47±1.16 58.10±1.86 58.52±1.50 0.69 NS

Means with different superscripts a, b in a row differ significantly (P < 0.05) NS- Non significant at 5% level * Significant at 5% level

4.6 NUTRITIVE EVALUATION IN TERMS OF PERCENT DIGESTIBLE

CRUDE PROTIEN (%DCP) AND PERCENT TOTAL DIGESTIBILE

NUTRIENTS (%TDN)

The nutritive evaluation in terms of percent digestible crude protein

(%DCP) and percent total digestible nutrient(%TDN) in complete feed

mixtures of ‘C’ , ‘T-1’ and ‘T-2’ in crossbred female calves has been given

Table 4.9.

50

The percent DCP values for ‘C’ , ‘T-1’ and ‘T-2’ diet groups( 6.52+ 0.20,

7.59+ 0.30 and 6.61+ 0.32) were not similar and varies statistically (P< 0.05).

The percent TDN values of ‘C’, ‘T-1’ and ‘T-2’ dietary groups were quite

similar (61.36+ 1.69, 62.81± 2.42 and 61.60±2.41)and did not vary statistically

(P< 0.05). The data showed that the ‘T-1’ diet had higher DCP than ‘C’ and ‘T-

2’ diets indicating higher availability of DCP in ‘T-1’ diet but the energy

availability remained the same as the percent TDN remained the same due to

similar digestibility of nutrients in all the groups.

Thus, the highest availability of digestible protein in ‘T-1’ diet did not

reflected in the body weight gain in this group of crossbred female calves

indicating that DCP utilization was not efficient in ‘T-1’ diet.



nutritive evaluation of ration in female crossbred calves

Particulars SBM ‘C’

CSC ‘T-1’

DCSC ‘T-2’

SEm

%DCP 6.52a±0.20 7.59b±0.30 6.61a±0.32 0.20*

%TDN 61.36±1.69 62.81±2.42 61.60±2.41 1.20 NS

Means with different superscripts a, b in a row differ significantly (P < 0.05) NS- Non significant at 5% level

4.7 NITROGEN BALANCE

The effect of partial replacement of soybean meal ‘C’ with cottonseed cake or

decorticated cottonseed cake solvent extracted in ‘T-1’ or ‘T-2’ diets on the

nitrogen balance in crossbred female calves have been shown in Table 4.10.

51

Table 4.10 Effect of partial replacement of soybean meal with cottonseed

cake (expeller) or decorticated cottonseed cake (Solvent extracted) on

Nitrogen balance (g) in female cross bred calves

Particulars

Groups SEm

(P

<0.05) SBM

‘C’

CSC

‘T -1’

DCSC

‘T -2’

Nitrogen intake (g/d) 84.08±5.80 81.23±5.99 80.00±4.60 3.30NS

N outgo through

faeces(g/d) 36.98b±4.48 33.23b±2.99 25.46a±4.53 2.23*

N outgo through urine(g/d) 16.15±3.11 19.22±3.53 17.24±3.93 1.94NS

Total outgo (g/d) 53.13±7.42 52.45±5.99 42.70±8.35 3.97NS

N Balance (g/d) 30.95ab±2.61 28.78a±1.14 37.30b±5.61 2.42*

Absorb N(g/d) 47.10ab±2.50 48.00a±3.74 54.28b±3.35 2.21*

Percent Absorb N 56.67a±2.97 59.15a±2.19 66.01b±3.80 1.78*

N Retained 38.21±4.98 36.39±2.99 46.39±7.01 3.00NS

N Retained (%) of N

absorbed 66.22±5.61 61.63±4.74 69.32±7.31 3.37NS

Means with different superscripts a, b, ab in a row differ significantly (P < 0.05) NS- Non significant at 5% level * Significant (P < 0.05)

The nitrogen balance was measured as the nitrogen intake through

feed, nitrogen outgo through faeces and nitrogen outgo through urine. Total

nitrogen intake (g/d) through feed was 88.08±5.80, 81.23±5.99 and

80.00±4.60 respectively in three dietary groups ‘C’, ‘T-1’ and ‘T-2’. Nitrogen

outgo through faeces per day was 36.98± 4.48, 33.23±2.99 and 25.46±4.53.

the nitrogen outgo through urine (g/d) was obtained to be 16.15±3.11,

19.22±3.53 and 17.24±3.93 and the nitrogen balance of calculated as

52

30.95±2.61, 28.78±1.14 and 37.30±5.61 in three groups of concentrate

mixtures ‘C’, ‘T-1’ and ‘T-2’ respectively.

The total nitrogen outgo was low in ‘T-2’ diet (42.70±8.35) compared to

‘C’ and ‘T-1’ group but the nitrogen balance was found to be higher in ‘T-2’

diet; which also reflected in higher body weight gain in this group (Table 4.6).

Though non significantly the nitrogen retained as percent of nitrogen

absorbed was also higher in ‘T-2’ group indicating a higher biological value of

this ‘T-2’ diet compared to the other two dietary groups i.e. ‘C’ and ‘T-1’.

Khalid et al. (2012) observed that organic nitrogen sources with high RUP

(Rumen undegradable protein) were more efficient in improving the nitrogen

balance in lambs when compared to the lower RUP or inorganic nitrogen

source. Increased nitrogen retention was observed in lambs fed diets

supplemented with CSM as a natural higher undegradable protein source

(Caton et al., 1988).

4.9 BLOOD BIOCHEMICAL PROFILE

The partial replacement of soybean meal with cottonseed cake

(expeller) or decorticated cottonseed cake (Solvent Extracted) showed no

adverse effect on blood biochemical profile (4.11). Blood urea nitrogen values

(mg/dl) were significantly higher in ‘T-2’ (18.08±0.64) compared to other

(14.77±0.61, 14.67±0.58). The values for blood glucose (mg/dl) 60.19±1.15,

60.15±0.89 and 59.30±0.95, total protein (g/l) 7.13±0.23, 7.38±0.27 and

7.51±0.26, albumin (g/l) 3.72±0.10, 3.86±0.07 and 3.66±0.11, globulin (g/l)

3.42±0.23, 3.52±0.27 and 3.81±0.28, Albumin: Globulin ratio 1.26±0.10,

1.45±0.19 and 1.20±0.13 and haemoglobin (g/dl) 11.11±2.63, 8.60±0.22 and

8.30±0.25 in control, T-1 and T-2 respectively, did not vary much and

remained in normal range. Numerically the values for glucose were found to

be little lower in ‘T-1’ and ‘T-2’ diet compared to ‘C’ may be due to gossypol in

cottonseed cakes.

53

Table 4.11:- Effect of partial replacement of soybean meal with


blood biochemical profile in female cross bred calves

Particulars

Groups SEm

(P <0.05) SBM

‘C’

CSC

‘T -1’

DCSC

‘T -2’

BUN (mg/dl) 14.77a±0.61 14.67 a±0.58 18.08b±0.64 0.39*

Glucose (mg/dl) 60.19±1.15 60.15±0.89 59.30±0.95 0.57 NS

Total protein (g/l) 7.13±0.23 7.38±0.27 7.51±0.26 0.15 NS

Albumin(g/l) 3.72±0.10 3.86±0.07 3.66±0.11 0.05 NS

Globulin (g/l) 3.42±0.23 3.52±0.27 3.81±0.28 0.15 NS

Albumin : Globulin 1.26±0.10 1.45±0.19 1.20±0.13 0.08 NS

Haemoglobin (g/dl) 11.11±2.63 8.60±0.22 8.30±0.25 0.88 NS

Means with different superscripts a, b in a row differ significantly (P <0.05) NS- Non significant at 5% level * Significant (P <0.05)

The blood profile of cattle given cottonseed @1.2-2.0kg/d showed

gossypol toxicity with higher values of haemoglobin, proteins, albumin:

globulin ratio, urea and inorganic phosphorus (Mena et al., 2004).In contrast

Torane et al. (2006) reported significantly lower (P<0.01) concentration of

BUN in calves fed experimental ration T2 containing urea treated wheat straw

and un-decorticated cottonseed cake. Cottonseed cake is a source of natural

rumen protected protein and is an indicative of slow release of ammonia in the

rumen from such type of naturally protected slowly degradable protein source.

Colin et al. (1996) showed no significant change in the blood glucose and uric

acid levels in cattle fed whole cottonseed meal for 430 days.

Blood urea nitrogen (BUN) is a measure to assess protein status of

animal. Blood urea nitrogen and protein intake has a positive relationship and

54

BUN value is an indicator of protein intake (Preston et al. 1965; Rusche et al.

1993). It has also been reported that plasma glucose and urea N were

unaffected in lambs fed concentrate diets (Carro et al., 2006) while in contrast

to this Sano et al. (2007) found that increased in glucose concentration may

be due to more by-pass protein and increased availability of glucogenic amino

acids for glucose synthesis. Similarly Khalid et al. (2012) observed that

feeding protein with high rumen un-degradable value resulted in increased

concentration of blood glucose due to more glucogenic amino acids available

for gluconeogenesis. However, this could not be confirmed in the present

study.

Preston et al. (1965) reported that the quantity of ammonia absorbed

from the rumen was reflected in circulating BUN. Several studies (Roseler et

al., 1993; Baker et al., 1995; Hong et al., 2003) had shown that MUN (milk

urea nitrogen) was highly correlated with BUN. In healthy ruminants, BUN and

MUN concentrations are indicative of the protein to energy ratio in the diet.

Decreased haemoglobin and TEC are symptomatic of gossypol toxicity.

Lindsey et al. (1980) found depressed haemoglobin, and total protein of

plasma got elevated in cows fed the solvent meal. While Nikokyris et al.

(1991) could not observe any effect on haemoglobin in lambs when fed diet

incorporated with cottonseed meal up to 30%.

4.10 ECONOMICS OF FEEDING

The economics of feeding cottonseed cake or decorticated cottonseed

cake solvent extracted as a partial replacement of 10 unit of soybean de-oiled

in the concentrate mixture (4.12) have been calculated by considering the

market purchase prices of feed ingredients, oat fodder and wheat straw

(Table 4.1). The values regarding the economics of feeding and body weight

gains of animals are presented in Table 4.12. The cost of concentrate

mixtures was worked out to be Rs 1660, 1507 and 1512 per/100kg in ‘C’, ‘T-1’

and ‘T-2’ diets (4.12).

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55

4.12 Economics of feeding

Attributes

Groups

SBM ‘C’

CSC ‘T-1’

DCSC ‘T-2’

Cost of concentrate (Rs/100kg) 1660 1507 1512

Cost of wheat straw (Rs/100kg) 300 300 300

Cost of green oats (Rs/100kg) 150 150 150

Total intake of wheat straw (kg)* 150.4 134.9 144.66

Cost on wheat straw (Rs) 451.20 404.7 433.98

Total intake of concentrate (kg)* 201.8 182.6 192

Cost on concentrate (Rs) 3349.88 2751.78 2903.04

Total intake of green oats (kg)* 736.25 731.25 763.13

Cost on green oats (Rs) 1104.37 1096 1144.69

Total cost of feeds (Rs) 4905.45 4253.35 4481.72

Total weight gain (kg) 66.60 61.30 78.80

Cost /kg gain 73.65 69.38 56.87

*105 days

The differences in cost of concentrate mixtures were due to

replacement of costly soybean meal with little cheaper cottonseed cake

(expeller) or decorticated cottonseed cake (solvent extracted) Table 4.1. The

market price of green oats was Rs 150/qtl, and that of wheat straw was taken

as Rs 300/qtl. The cost of total feed intake for 105 days also varied in different

groups due to variation in feed intakes and the cost worked out was Rs 4905/-

, Rs 4235/- and Rs 4481/- for ‘C’, ‘T-1’ and ‘T-2’ diets respectively, showing

lower cost for ‘T-1’ and ‘T-2’ diets because both cottonseed cake and

decorticated cottonseed cake were cheaper than soybean meal (de-oiled).

The total gain in experimental animals during a period of 105 days was 66.60,

61.30 and 78.8 kg in ‘C’, ‘T-1’ and ‘T-2’ diets respectively.

The net cost of production i.e. per kg gain in body weight was Rs

73.65, 69.38 and 56.87 in ‘C’, ‘T-1’ and ‘T-2’ diets respectively indicating that

56

‘T-1’ and ‘T-2’ concentrate mixtures replaced with cottonseed cakes (Expeller

and Solvent Extracted) were better and economical than soybean meal de-

oiled.

CHACHACHACHAPTER PTER PTER PTER –––– 5 5 5 5

SUMMARY &CONCLUSIONSUMMARY &CONCLUSIONSUMMARY &CONCLUSIONSUMMARY &CONCLUSIONSSSS

57 Summary & ConclusionsSummary & ConclusionsSummary & ConclusionsSummary & Conclusions

SUMMARY AND CONCLUSION

An experiment was conducted to see the effect of partial replacement

of soybean meal (SBM) with cottonseed cake (CSC) or decorticated

cottonseed cake (DCSC) on growth, nutrient utilization, body weight gain in

growing calves and on blood biochemical profile. Also the in vitro protein

degradability of decorticated cottonseed cake (Solvent extracted), cottonseed

cake, (expeller) soybean meal (de-oiled), Ground Nut cake (expeller) and

other ingredients of the concentrate mixture and also the experimental

concentrate mixture was measured. The control ‘C’ diet contained 15%

soybean meal and 10 parts of this was replaced by cottonseed cake (expeller)

in group ‘T-1’ diet and decorticated cottonseed cake (solvent extracted) in

group ‘T-2’ and concentrate mixtures were prepared.

The rumen in vitro dry matter, organic matter, crude protein

degradability (1st stage Tilley and Terry,1963) was measured for soybean

meal 64.41±1.61, 64.49 ±0.58 and 64.46±0.60, cottonseed cake (expeller)

59.58±1.02, 52.86 ±1.58 and 58.96 ±1.98 and decorticated cottonseed cake

(solvent extracted) 55.70 ±0.45, 53.31±0.70 and 55.92 ±0.87, respectively.

Also for the control ‘C’ concentrate mixture 71.41±1.26, 67.03±0.85 and

65.74±1.73, conc. ‘T-1’ 67.92±1.10, 58.28±0.94 and 59.04±1.4 and conc. ‘T-2’

65.05±0.93, 55.03±0.91 and 56.54±1.25, respectively.

The animals were fed ad libitum (concentrate and wheat straw 50:50

and 10kg green oats to full fill the requirement of vitamin A) and clean tap

water provided to the animals every day. Daily feed intake and fortnightly

body weight of the calves were recorded for 105 days. After completion of 85

days of experimental period, a metabolic trial of 7 days duration was

conducted on all the animals to assess their voluntary feed intake, productive

performance, nutrient digestibility, nitrogen balance and the nutritive

evaluation of the feed given to the different group of the animals, concentrate

mixture ‘C’, ‘T-1’ and ‘T-2’ contained crude protein (%)19.1, 20.0 and 19.9,

ether extract (%) 4.2, 6.2 and 4.8 and nitrogen free extract (%) 56.3, 52.0 and

55.8 respectively and contributed towards meeting the needs of nitrogen as

well as readily available source of energy to the animals for rapid growth.


Wheat straw contained 90.6, 2.6, 33.1, 0.9, 45.8, 81.6, 57.1 and 24.5 percent

of DM, CP, EE, Nitrogen free extract (NFE), NDF, ADF and hemicellulose

respectively. Green oats contained 16.0, 8.5, 2.7, 44.2, 63.2, 43.0 and 20.2

percent of DM, CP, EE, Nitrogen free extract (NFE), NDF, ADF and

hemicellulose respectively.

At the start of experiments, the average body weights of the groups

were, group ‘C’ 114.37±20.35 kg, group ‘T-1’ 13.17±23.13 kg and group ‘T-2’

115.37±26.96 kg respectively. The average live weight of the cross bred

(Karan fries) female calves at the end of 105 days of experimental period

were ‘C’ 181.00 ±23.63 kg, ‘T-1’ 174.50±28.17 kg and ‘T-2’ 194.17±34.19kg,

respectively. Thus the calves of ‘T-2’ shows significant (P≤0.05) higher gains

in body weights (78.8±10.32 kg) than ‘C’ and ‘T-1’ diet (66.6±3.96 and

61.3±6.50 kg) respectively. Also the average daily gains were observed to be

significant (P≤0.05) higher in ‘T-2’ (750.0±51g) than ‘C’ and ‘T-1’ (634.0±61

and 583.0±37 g).

The results further showed that the total DMI for 105 days

experimental period (434.9±42.00, 402.8±46.30 and 425.1±41.40kg) DMI per

day (4.14±0.40, 3.83±0.44 and 4.04±0.39 kg), feed conversion ratio

(6.26±0.52, 6.75±1.80 and 6.69±1.07) and percent feed efficiency

(16.21±2.13, 15.34±0.69 and 18.55±1.73) in ‘C’, ‘T-1’and ‘T-2’ diet, did not

differ significantly in three groups.

The digestibility coefficient (%) of dry matter (63.63±1.32, 61.19±1.10

and 60.62±1.63), organic matter (69.35±1.38, 67.42±1.40 and 66.97±1.47),

crude protein (59.77±2.51, 59.7±1.93 and 61.00±2.73), crude fibre

(64.51±1.70, 58.87±1.25 and 61.69±2.44), nitrogen free extract (70.24±1.20,

65.84±1.50 and 70.13±1.90), acid detergent fibre (50.93±1.86, 48.96±1.49

and 46.08±2.71) and neutral detergent fibre (60.47±1.16, 58.10±1.86 and

58.52±1.50) in ‘C’, ‘T-1’and ‘T-2’ diet respectively, the values were not

significantly different in three groups. However the digestibility coefficient for

ether extract was significantly higher in group ‘T-1’ and ‘T-2’, (87.16±1.04 and

86.39±0.93) than group ‘C’ (79.34±1.43) diet.

Nitrogen intake (g/d) was 84.08±5.80, 81.23±5.99 and 80.00±4.60 in

‘C’, ‘T-1’ and ‘T-2’ groups respectively. Nitrogen intake did not differ

significantly among the three groups (P≤0.05). Excretion of nitrogen (g/d)


through faeces was lower in group ‘T-2’ 25.46±4.53 than ‘C’ and ‘T-

1’(36.98±4.48 and 33.23±2.99) and through urine was (16.71±3.11, 19.22

±3.53 and 17.24±3.93) in ‘C’, ‘T-1’ and ‘T-2’ diet groups respectively, which

did not differ significantly among the three groups (P≤0.05). However, it was

observed that nitrogen balance (g/d) was significantly higher (P≤0.05) in

group ‘T-2’ (37.30±5.61), than group ‘C’ and ‘T-1’(30.95±2.61 and 28.78±1.14)

and absorbed nitrogen was also significantly higher in ‘T-2’ (54.28±3.35) than

‘C’ and ‘T-1’ (47.10±2.50 and 48.00±3.74). Thus animals in ‘T-2’ dietary

group showed higher percent of absorbed nitrogen (66.01±3.80) than ‘C’ and

‘T-1’ diets i.e.(56.67±2.97 and 59.15±2.19). All the animals were found in

positive nitrogen balance.

The blood urea nitrogen was found significantly higher in ‘T-2’

(18.08±0.64) than ‘C’ (14.77±0.61) and ‘T-1’ (14.67±0.58), besides the BUN,

values of glucose, total protein, albumin, globulin, albumin: globulin ratio and

haemoglobin varied slightly but not significantly in all three groups.

The total input cost (Rs) on different dietary treatments ‘C’, ‘T-1’ and ‘T-2’

was 4905.45, 4253.35 and 4481.72, total gain in body weight (kg) 66.60, 61.3

and 78.8 and cost/kg gain (Rs) 73.65, 69.38 and 56.87in ‘C’, ‘T-1’ and ‘T-2’

groups respectively.

It can be concluded that incorporation of decorticated cotton seed

cake as a partial replacement (10 parts) of soybean meal in a concentrate

mixture fed as a complete feed mixture will show higher growth rate in

crossbred female calves with better percent feed efficiency and no effects on

blood profile.

As the market price of decorticated cotton seed cake (Solvent extract)

is less than soybean meal de-oiled and it showed better growth and average

daily gains than soybean meal. Thus it will be in advantage to feed

decorticated cottonseed cake solvent extracted in a concentrate mixture for a

rapid rate of growth in crossbred female calves.

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EFFECT OF FEEDING DECORTICA TED COTT ONSEED CAKE …...Ishtiyak sir, Vijay sir, Nazam sir, Sanjay...

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