EFFECT OF SUPPLEMENTAL NEEM LEAF MEAL ON

PERFORMANCE OF BROILER BIRDS,

BY

OLABODE ADEYEMI DAVID

PG/M.Sc/05/39487

A research project report submitted to the department of

Animal Science, Faculty of Agriculture, University of

Nigeria, Nsukka; in partial fulfillment of the requirement

for the award of the degree of Master of Science (M.Sc) in

Animal Nutrition.

FEBUARY, 2009.

I

CERTIFICATION

We certify that Mr. OLABODE ADEYEMI DAVID, a postgraduate

student in the department of Animal Science with the registration number

PG/M.Sc/05/39487, carried out this research work for the award of the

degree of Master of Science in Animal Nutrition. The work embodied in

this project report is original and has not been previously reported

elsewhere.

……………………………… ……………………………

Prof. G.C. Okeke Dr.A.E. Onyimonyi

(Supervisors)

………………………………..

(Head of Department)

……………………………….

………………………………

(External Examiner) (Internal Examiner)

II

DEDICATION

This work is dedicated to my beloved Wife, Favour Olabode and my late

father Mr. A. O. Olabode (FAAI).

iii

ACKNOWLEDGEMENT

My profound gratitude goes to my project supervisors; Prof. G.C. Okeke

and Dr.A.E. Onyimonyi, for their thorough supervision of this work, which

has made it what it is today. I shall not fail to thank Dr. Ezekwe and other

members of the academic staff and non- academic staff of the

department for their assistance throughout my study.

I wish to express my sincere gratitude to my wife, Olabode Favour

who was of great help and assistance to the completion of this project

work.

Special thanks to my pastor, Pastor Afanga Saviour (Winners Chapel,

Okigwe, Imo State) for his unceasing prayers for me and every member of the

Winners family Okigwe. My thanks also goes to Pastor John Olabode for his

support and encouragement. May the good Lord reward and bless him greatly.

Olabode Adeyemi D.

iv

TABLE OF CONTENTS

CONTENT PAGE

Title page i

Certification ii

Dedication iii

Acknowledgement iv

Tables of content v

List of tables vii

Abstract viii

CHAPTER ONE

Introduction 1 - 4

Objectives 4

CHAPTER TWO

Literature Review

2.1 Poultry Nutrition 5 - 6

2.2 Nutrient Requirement for Broilers 6 - 7

2.2.1 Energy Requirements 7 - 8

2.2.2 Protein and Amino acid Requirement 8 - 10

2.2.3 Water Requirement 10 - 11

2.2.4 Mineral Requirement 11 - 12

2.2.5 Vitamin Requirement 12 - 13

2.3.1 Characteristics and features of leaf meals. 13 - 14

2.3.2 Utilization of leaf meals in poultry. 14 - 16

2.3.3 Neem tree leaves. 16

2.4 Hematological studies. 17 - 18

V

CHAPTER THREE

Materials and Methods

3.1 Location and Duration of study 19

3.2 Experimental Design 19

3.3 Processing of Neem leaves 19 - 20

3.4 Experimental Diets 20 - 22

3.5 Management of Birds 23

3.6 Response Parameters 23

3.7 Statistical Analysis 24 - 25

3.8 Laboratory Analysis 25

CHAPTER FOUR

Result and Discussions

4.1 Average daily weight gain. 27 - 28

4.2 Average daily feed intake. 28 - 29

4.3 Feed conversion ratio 29

4.4 Hemoglobin (Hb) values. 30 - 31

4.5 Packed cell volume (PCV) values. 31

4.6 Aspartate aminotransferase (AST) values. 31 - 32

4.7 Bilirubin values. 32

CHAPTER FIVE

5.1 Summary and Conclusion 33 - 34

5.2 Suggestion 35

5.3 References 35 - 41

5.4 Appendices 42 - 44

vi

LIST OF TABLES

1. Amino acid requirements by broilers (%) 10

2. Composition of Broiler starter diets supplemented with graded levels

of neem leaf meal. 21

3. Composition of Broiler finisher diets supplemented with graded levels

of neem leaf meal. 22

4. Proximate composition of processed Neem leaf meal (NLM) 26

5. Effects of varying levels of supplemental NLM on the performance of

broiler birds. 27

6. Effects of supplemental NLM on the Hematological values

of broiler birds. 30

vii

ABSTRACT

Ninety “Ross” unsexed day old broiler chicks were used to determine

the optimum level of neem leaf meal supplementation in broiler diet. The

experiment was laid out in a completely randomized design (CRD). The birds

were randomly selected and divided into five treatment groups of eighteen birds

each. Each treatment group had two replicates with nine birds per pen. The birds

were fed five diets containing neem leaf meal at 0, 0.5, 1, 1.5 and 2 % levels of

inclusion. Results revealed that feed intake of birds on diet 1 (129.39g), 2

(131.71g) and 3 (128.24g) were similar (P>0.05) but were significantly (P<0.05)

higher than those of birds fed diet 4 (126.56g) and 5 (110.60g). Similarly weight

gains of birds on diet 1 (40.82g), 3 (40.20g) and 4 (39.80g) did not differ

significantly (P>0.05) but birds on diet 2 (44.80g) had a higher weight gain

which resulted in a significantly higher final body weight. Hemoglobin (Hb)

values of birds fed diet 2 (10.8g/dl) and 5 (10.2g/dl) were similar (P>0.05) but

significantly (P<0.05) higher than those of birds fed diet 1 (8.9g/dl), 3 (8.8g/dl)

and 4 (9.0g/dl). Also the packed cell volume (PCV) values of birds fed diet 2

(39%) was significantly (P<0.05) higher than those fed diet 1 (32%), 3 (30%), 4

(34%) and 5 (36%) respectively. The overall result shows that birds on diet 2

(0.5%, NLM) had significantly (P<0.05) superior body weight gain and final

body weight than birds on diet 1 (0%, NLM), 3 (1%, NLM), 4 (1.5%, NLM) and

5 (2%, NLM). Also hematological values investigated were within accepted

normal range for poultry, except for hemoglobin where diet 1 and 3 fell slightly

below the recommended values for broiler birds.

ix

CHAPTER ONE

INTRODUCTION

Nutrition is the most important consideration in any livestock

enterprise. Its (livestock industry) survival is dependent on the

availability of feedstuffs, which are mainly components of human food

(Esonu et al., 2006). The aim of keeping livestock and poultry is for

the production of high quality protein. Agriculturists and Nutritionist

in Nigeria have generally agreed that poultry is the fastest means of

bridging the protein deficiency gap prevailing in the country (Olomu,

1995; Atteh, 2002).

According to Olomu (1995) and Kekeocha (1994), broiler birds

are fast growing birds and are described as good converters of feed

and are marketed from eight to twelve weeks. Broiler birds are

regarded as the type of birds that have high feed consumption and

conversion ratio of non-conventional feed ingredients that cannot be

directly consumed by man into high quality meat which are needed in

large quantity by man (Partmouth, 1991). However, the production of

broiler birds as meat bird cannot be effective in the absence of

adequate feed and feed ingredients in right proportion. It has been

estimated that 70-80% of poultry production inputs is been attributed

to feed cost, hence making it imperative that good quality feed be

available at reasonable price to make poultry production more

profitable (Oruwari et al., 1995).Thus, the birds can only perform

economically well and profitably if it consumes, on daily basis, the

appropriate amount of energy, protein, vitamins and minerals (Richard

and Church, 1998).

1

Antibiotics are drugs such as penicillin, amoxicillin and

tetracycline that are used to kill or inhibit the growth of harmful

bacteria (Atteh, 2002). Thus, the use of antibiotics in improving the

rations of birds have been carried out by various authors. Onifade and

Babatunde (1995) reported that growth depression in broiler chickens

fed high fibre diet was effectively and economically alleviated by the

action of the following antibiotics namely tylosin, streptomycin and

neoterramycin. Also Onifade and Babatunde (1996) reported that the

growth performance of broiler chicks fed a high fibre diet

supplemented with antibiotics was better than those fed

unsupplemented diet. Thus, antibiotics supplementation is an effective

and readily adaptable strategy, if not for the possibility of their

immunosuppressive actions under prolonged usage.

However the inclusion of antibiotics in livestock rations as feed

additives is being discouraged because of the negative effect on the

animals and the humans who consume the final animal products

(HCWH, 2005). This was also confirmed by Oyekunle and

Owonikoko (2002). Dipeolu (2002) and Chan et al. (2002) who

reported problems of residual effect in livestock products. Thus, there

is the need to evaluate potential antibiotic alternatives to improve

disease resistance in intensive food animal production. Nutritional

approach to counteract the debilitating effects of stress and infection

may provide products with useful alternatives to antibiotics.

Improving the disease resistance of animals grown without antibiotics

can benefit the animal’s health, potentially increasing production

efficiency and food safety. Improving disease resistance in food

2

animals particularly in the absence of antibiotics treatment is a key

strategy in the effort to increase food safety (HSUS, 2007).

Neem tree (Azadirachta indica) is an indigenous tropical plant

predominant in Nigeria. It is known by names such as “Ogwu Iba” in

Ibo land and “Dogonyaro” in Hausa. Neem tree is a member of the

family meliaceae in the order fagates of the Dicotyledonous class in

the plant kingdom. When fully grown, it can reach 24m in height with

dense wide spreading crown. The crown bears large number of

compound leaves each with 5-9 pairs of leaflets. Flowering is twice a

year (April/May and August/ September). When fertilized one seeded

fruits are produced with fibrous cover. The tree can yield between 20 –

50kg of fruits annually (Aliero, 2003).

The neem has been reported to contain several biologically

active constituents such as azadirachtin, meliantriol, salanin as well as

nimbin and nimbidin (Schmutterer, 1990). He also reported salanolide

(a meliacin) as one of the bitter principles in neem seed oil.

Azadirachta indica is medicinal and it is used as an insecticide and

pesticide. The tree is relevant in organic farming. The leaf meal has a

proximate composition of 92.42% dry matter; 7.58% moisture;

20.68% crude protein; 7% ash; 4.13% ether extract; 16.60% crude

fibre; and 43.91% nitrogen-free extract (Esonu et al., 2005). The

nutritive value of the neem leaf meal has also been reported by the

same author. They observed that carcass weight, dressed weight, liver,

heart and gizzard weights of laying hens fed varying levels of neem

leaves were significantly increased at 5 per cent level. There is

however scarcity of information in the literature on the utilization of

neem leaves by broilers. However, factors such as nutrient imbalance,

3

improper metabolism, presence of anti-nutritional factors and toxic

elements in such novel feed ingredient have been implicated in similar

productions.

OBJECTIVES OF THIS STUDY

i. To evaluate the performance of finishing broilers fed graded

levels of neem leaf meal.

ii. To investigate the hematology and blood chemistry of

finishing broilers fed graded levels of neem leaf meal.

4

CHAPTER TWO

LITERATURE REVIEW

2:1 POULTRY NUTRITION

The nutrition of animal involves various activities that result in

the conversion of feed into animal tissues or animal products for

example egg, meat, milk e.t.c (Richard and Church, 1998). Pond et al,

(1995) defined nutrition as a series of processes by which an organism

takes and assimilates food for promoting growth and replacing worn-

out tissues. Also Obioha (1992) defined nutrition as a process which

provides nourishment to a living organism and thus the food which a

living organism takes in is used primarily for two major purposes;

maintenance and productive function.

Olomu (1995) emphasized that poultry nutrition is more critical

than other farm animals (except pig) because poultry birds are usually

reared in confinement. Poultry are more active and more sensitive to

environmental influences than other farm animals. According to

Obioha (1992), poultry have high growth rate, intensive metabolic

rates and rapidly developed reproductive organ. He further reported

that during the first ten weeks of post embryonic growth, the weight of

broilers increases for about thirty to forty times and that such rate of

growth have not been encountered in other farm animals.

The ultimate aim of poultry nutrition is to increase the

productive efficiency of the poultry. The economic importance of

poultry feeding has become apparent, when it was realized that feed is

the highest single cost factor, about two third of the total production

cost . Many cases of poultry business failures can be traced to poor or

5

improper feeding of the birds. Therefore, it is pertinent that the right

nutrients in adequate quantities must be supplied to the birds.

According to Richard and Church (1998), quality of ingredients as

well as quality of nutrients present must be evaluated.

2.2 NUTRIENTS REQUIREMENT FOR BROILERS.

On the basis of physiological structures of the poultry, a good

knowledge of their nutritional requirement is very important. Thus,

lack of understanding of the digestive system of birds could put the

producer in a serious problem. Nutrient can be defined as chemical

substances found in feed ingredients which when made available to the

animal are used for maintenance, production and health of the animals

(Atteh, 2002). It can also be described as a specific chemical element

or compound supplied by or derived from the diet and absorbed into

the body tissues to support physiological processes. These nutrients

are essential to be in appropriate amounts and proportions in the diets

of intensively housed birds, because they have no access to other

sources of nutrients. The essential nutrients required by the poultry

birds are carbohydrates, fat and oil (mostly served as energy givers);

protein (amino acid), minerals (micro and macro), vitamins and water

(Pond et al., 1995; Olomu, 1995 and Smith, 2001).

The dietary requirement of nutrients varies with age, bodyweight

and rate of growth of the birds. Thus the requirement for any nutrient

may be defined as the amount of that nutrient which must be supplied

in the diet to meet the needs of the normal healthy animal given on

otherwise completely adequate diet in an environment compatible with

good health (Olomu, 1995). Such level of nutrient must meet the

6

requirement for maintenance, growth, productive and reproductive

potential of the animal (NRC, 1994).

2:2: 1 ENERGY REQUIREMENTS

Poultry, like other farm animal, consume feed primarily to

satisfy their energy requirement. According to Obioha (1992), the

energy required by the bird to perform its maintenance and productive

functions is usually given to the animal through carbohydrate and fats.

Protein can also be broken down to supply energy, though this only

occurs where the lipid and carbohydrate sources of energy are

inadequate.

Cereal grains, cereal by-products, fats and oils from animal and

vegetable supply most of the energy in poultry diets. Poultry have

limited ability to utilize high fibrous feeds such as roughages. Olomu

(1995) reported that most chickens have the ability to adjust feed

intake in-order to obtain the necessary energy required for optimum

performance. When fed ad-libitum, chickens on low energy diets

consumed more feed than those fed high energy diets. Therefore, the

amount of nutrients required in the poultry rations must be adjusted in

relation to the energy levels of the rations.

Fats can be used to increase the energy level of low-energy

rations. Fats inclusion should be limited to not more than five to ten

percent of the diets. Fats supply about 2.5 times as much energy as

that which an equal weight of carbohydrate supplies (Pond et al.,

1995). It increases the utilization of feed and performs important

functions in the cells. In hot climate areas, feed containing added fats

might become rancid quickly, unless it has been properly stabilized. In

7

poultry, energy of the rations is usually expressed in unit of

metabolizable energy per unit weight; joules per kg, or calories per kg,

where one calorie is equivalent to 4.16 joules.

According to NRC (1994), the energy requirements of tropical

countries are lower than those of temperate regions. The recommended

energy requirements for chickens, pullets and layers under tropical

environments have been estimated as 3200; 2900 and 2900kcal/kg

respectively. Olomu (1995) fed birds’ dietary energy levels of 2500,

3000 and 3200kcal ME per kg. He observed that, the best performance

of chicks was realized with the ration containing 3000KcalME per kg.

For tropical environment, Obioha (1992) recommended an energy

level of 2850Kcal ME per kg for broiler starter diet and 2900Kcal ME

per kg for broiler finisher diet. It must be noted that increasing the

dietary energy resulted in an increase in the cost of diet.

2:2:2 PROTEIN AND AMINO ACID REQUIREMENT

Amino acids are the basic fundamental structural units of

protein, which are required by the birds for growth and for repairing

worn out tissues (Smith, 2001). Chickens and pigs can produce in their

bodies some but not all the amino acids. Those amino acids that must

be supplied in the feed are known as the essential amino acids. Thus

Smith (2001) reported that the protein requirement of a bird can be

defined as the birds’ requirement for the supply of each essential

amino acid together with sufficient supply of suitable nitrogenous

compounds from which the non-essential amino acids can be

synthesized. 8

According to Olomu (1995), under normal circumstance birds

eat more as they grow older. Therefore, the total protein consumed

increases as the birds gets older and presumably increase in weight.

However, the protein consumed per unit weight either reduces or

remains constant. Thus, if dietary protein level remains constant

throughout the life of the birds, more protein than necessary may

eventually be consumed. Since protein is not stored in the body to any

appreciable extent, any protein consumed above the birds’ requirement

is oxidized for energy (Olomu, 1995). All essential amino acids are

required by poultry and the most commonly deficient amino acids in

the chicken diets are arginine, lysine, methionine, tryptophan and non-

essential glycine (Obioha, 1992). Environmental temperature affects

protein requirements of chicks in the tropics. The high ambient

temperature in the tropics has been found to adversely affect the feed

intake as well as the protein intake of chicks. Olomu (1995), therefore

recommended that broiler starter diets should contain between 23 and

26% crude protein, if a maximum weight gain and efficient feed

utilization (good feed conversion) could be achieved. In terms of

amino acids requirements young birds have the highest demand for

essential amino acids as percent of the dietary protein (Oluyemi and

Robert, 1988).

9

Table 1: Amino Acid Requirement of Broilers (%)

Broiler Broiler

Starter Finisher

Lysine 1.3 1.1

Methionine 0.5 0.4

Leucine 1.5 1.4

Methionine + Cystine 0.75 0.7

Tryptophan 0.25 0.20

Isoleucine 0.85 0.80

Threonine 0.80 0.70

Arginine 1.10 1.0

Phenylalanine 0.80 0.70

Valine 1.0 0.85

Source: Olomu (1995).

In general, deficiency of essential amino acids in poultry diets leads to

poor growth, poor egg production and low degree of feed utilization.

2:2:3 WATER REQUIREMENTS.

Water is one of the most important nutrients needed by

broilers. Water constitutes more than half of the weight of poultry

meat and about two-third of the weight of egg (Ranjhan, 1981). The

newborn animal contains 750 to 800g of water per kg weight, though

this falls to about 500g/kg in the matured fat animal (McDonald et al.,

1995). Oluyemi and Roberts (2007) reported that for every kg of feed,

2-3 liters of water is consumed. Smith (2001) also reported that

drinking water for poultry should be free from salt and toxic

10

substances. He also reported that the ratio of dry matter to water intake

by poultry in a temperate environment is normally 1:2, but does

depend to some extent on the diet. Thus, increase in protein level also

increases the water consumption of the birds.

Water is available in the feed (metabolisable water) but the bulk

of water must be provided separately in drinking cans or troughs. The

water must be provided ad-libitum, clean and free from excessive

salts, which might have a laxative effect and must be cool. Olomu

(1995) suggested that 2 to 2.5 liters of water in the first two weeks is

adequate for 100 chicks, this is doubled after another 2 weeks. In

practice, the daily water consumption per 1000 broilers ranges from 20

liters at one week of age to 380 liters at 8weeks of age. On very hot

days, the water consumption may be increased as much as three times

of the water intake on cooler days.

2:2:4 MINERAL REQUIREMENTS

Minerals are mainly inorganic components of feed. The body

of animals contains large number of mineral elements, which occur in

combination with the organic constituents. Chicken body contains

about four per cent mineral matter (Ranjhan, 1981). Phosphorus and

calcium are the two most abundant mineral elements in the bone and

other skeletal components of the birds.

Olomu (1995) classified mineral elements as macro and micro

depending on the quantitative requirement for each. Macro elements

include; calcium, phosphorus, sodium, chlorine, sulphur, magnesium

and potassium while some of the micro-elements are iodine, zinc,

cobalt, iron, copper and molybdenum. Excess or deficiency of these

11

mineral elements is detrimental to the health and development of the

birds. Minerals function in many ways such as in body structural

components and acid-base balance. Calcium and phosphorus have

been found to have closely related metabolic functions. The

requirement of these minerals appears to be higher for warm climates

than for cold climates (Olomu, 1995). It is therefore recommended that

the level of calcium be increased from 1-1.2% to 4-4.5% and the level

of total phosphorous from 0.7-0.8% to 1-1.1% during periods of hot

weather. Ranjhan (1981) stated that mineral deficiency and over

supply have serious adverse effects and could lead to death of chicks.

Thus, in practice 0.30-0.35% common salt or sodium chloride would

take care of the requirements for sodium and chlorine which must be

supplied from sources other than the normal ingredients used in

formulating poultry feeds.

2:2:5 VITAMIN REQUIREMENTS

Vitamins are organic compounds that cannot be synthesized in

the poultry body and they are required in extremely small quantities

but absolutely essential for normal growth and health. Oluyemi and

Robert (2007) listed the vitamins required in poultry diets as follows:

the fat soluble- vitamins A, D,E and K and the water soluble vitamins-

thiamine (B1), niacin, riboflavin (B2), pyridoxine (B6), biotin,

pantothenic acid, folic acid, choline, and vitamin B12. Vitamin C can

be synthesized in the body of birds but they are required as antistress

in hot environment.

The deficiency of these vitamins in the feed consumed by birds or

non-availability of these vitamins to the birds usually result in disease

12

symptoms, such as rickets which is caused by vitamin D deficiency

associated with low or imbalance of calcium and or phosphorous level

in the diet; curled toe paralysis (Vitamin B12) and perosis associated

with deficiency of choline and biotin (Olomu, 1995). Deficiency of

folic acid causes poor feathering, poor feather pigmentation and

anemia (Obioha, 1992).

2:3:1 CHARACTERISTICS AND FEATURES OF LEAF

MEALS

Leaf meals have been observed as having potential in non-

ruminant nutrition and feeding especially in poultry production. The

nutrient contents of leaf meals, mostly those of the leguminous plants,

have relatively higher crude protein content than the non-leguminous

plants and cereals. Also leaf meal contains high level of crude fiber

when compared with other feedstuffs used in the livestock industry. In

most cases, the crude fiber content of leaf meal may equal or even

exceed the crude protein content (D’Mello, 1992), like in the case of

Cajanus cajan, Prosopis chillensis, Albizia falacata and Manihot

esculenta.

Thus, this tends to reduce the overall digestibility when there is

significant proportion of leaf meal in the diet as well as consequently

decrease in growth and egg production of birds (Tangendjaja et al.,

1990). Leaf meals also provide some essential vitamins such as

vitamin A and C, minerals and oxycarotenoids which cause yellow

color of broiler chicken skin, beaks, shanks and egg yolk (Opara,

1996). Lysine concentration in leaf meal is relatively higher than those

of grains and some by- products such as coconut oil meal (D’Mello

13

and Acamovic, 1989), but not as high as those of fishmeal or soybean

meal. They are deficient in sulphur containing amino acids though

their use can be enhanced on supplementation with methionine.

Except for cassava leaf meal, the metabolisable energy content

of leaf meals is generally low (D’Mello and Acamovic, 1989). This

therefore entails that leaf meals cannot fully replace high energy

ingredients in monogastric nutrition. Leaf meals contain carotenoids

which are a class of compounds which includes the carotenes; the

precursors of vitamin A and the xanthophylls which do not possess

vitamin activity of pigment.

The concentration of carotenoids in leaf meal however will

depend upon the duration and method of drying. Rapid drying of

Leucaena leucocephala foliage yielded a meal with carotenes and

xanthophylls concentrations of 484 and 932 mg/kg dry matter

respectively (Wood et al., 1983). However they further said that

substantial losses occurred during oven drying at 60oC and 145

oC.

They also reported that carotenoids were more stable in sun dried L.

leucocephala leaf than in oven dried samples.

2:3:2 UTILIZATION OF LEAF MEALS IN POULTRY

At a high inclusion level, leaf meals lead to depressed growth.

This may be attributed to their low digestibility and inadequate

metabolisable energy content. Egbewande et al. (2004) have reported

that a graded level of mistletoes (Tapinanthus bangwensis) leaves

induced dose-related depression in growth of chicks. Also with older

broiler chicks, D’Mello et al. (1987) showed that diets containing

100g leaf meal per kilogram diet significantly reduced growth without

14

affecting dry matter intake. Although feed intake remained unaltered,

the chicks failed to consume sufficient quantities of the digestible

nutrients particularly protein and energy required to sustain rapid

growth, they further reported.

D’Mello and Acamovic (1982) reported no appreciable

differences in weight gain or efficiency of feed utilization between

control chicks fed a maize-soybean meal diet and those offered diets

containing Leucaena leucocephala leaf meal. Two trials by Ranaweera

et al. (1981) involving feeding Gliricidia sepium leaf meal to

growing chicks at inclusion rates of 50g/kg and 100g/kg diet

respectively resulted to satisfactory growth rate relative to control

animals while chicks fed a higher concentration of the leaf meal

(150g/kg) had markedly reduced growth rates. Inclusion of Robinia

pseudoacacia leaf meal at the rate of 200g/kg diet depressed growth

of chicks to 71% of that observed in the control group fed a maize

soybean diet (Cheeke et al., 1983).

Leaf meal derived from Sesbania sesban and S. grandiflora were

highly toxic to chicks even at inclusion rates as low as 100g/kg diet

(Shqueir et al., 1989). They also recorded mortality of 100% but

reported that chicks fed alfalfa meal at the same level gained 442g

over a three weeks period without any record of mortality.

The inclusion of Gliricidia sepium leaf meal at rates of up to

100g/kg diet induced small increase in feed to gain ratio of chickens

but a striking increase in this ratio occured on inclusion of the leaf

meal at 150g/kg diet (Ranaweera et al., 1981). Ravindaran et al.,

(1986) observed that cassava leaf meal depressed feed conversion

efficiency particularly when the leaf was used to replace soybean and

15

maize. They however, also observed that when cassava leaf meal was

used in replacing a poor quality raw material such as coconut oil meal,

the feed conversion efficiency was not depressed until levels of the

leaf meal reached 300g/kg in diets for chicken.

2:3:3 NEEM TREE LEAVES

Neem is an attractive broad leaved evergreen tree which can

grow up to 24m tall and 2.5m in girth (Aliero, 2003). Its spreading

branches form a rounded crown of deep green leaves and honey

scented flowers as much as 20m across. The leaves mainly yield

quercetin (Flavonoid) and nimbosterol (β sitosterol) as well as a

number of liminoids (nimbin and its derivatives). Quercetin (a

polyphenolic flavonoid) is known to have antibacterial and antifungal

properties (Elangovan et al., 2000). Liminoids like nimocinolide and

isonimocinolide affect fecundity in house flies (Musca domestica) at a

dose ranging between 100 and 500ppm. Fresh matured leaves yield an

odorous viscous essential oil which exhibits antifungal activities

against fungi (Trichophyton mentagrophytes) in vitro.

Few works have been carried out using neem tree leaves in

livestock production. Among such work include those carried out by

Esonu et al. (2006) who reported that up to 15% neem leaf meal in

treatment diet may increase both hen day egg production and egg yolk

color. Bawa et al. (2007) reported that up to 20% level of neem seed

cake can replace groundnut cake in a young rabbit ration. Uko and

Kamalu (2007) have also shown that autoclaved or toasted neem seed

kernel can be used to improve the carcass characteristics of broilers.

16

2:2:4 HEMATOLOGICAL STUDIES

Hematology and Serum biochemistry assay of livestock suggests

the physiological disposition of the animals to their nutrition. Esonu et

al. (2001) had stated that hematological constituents reflect the

physiological responsiveness of the animal to its internal and external

environments which include feed and feeding. Awosanya et al. (1999)

have observed the dependence of blood protein and creatinine on the

quality of dietary protein while, Iyayi (2001) feeding swine with

cassava leaf supplement found that serum glutamic oxaloacetic

transaminase (SGOT) was significantly lowered while serum total

protein was significantly increased.

The Aspartate aminotransferase test measures the level of

aspartate aminotransferase (AST), an enzyme released into the blood

when certain organs or tissues, particularly the liver and heart are

injured. Aspartate aminotransferase (AST) is also known as serum

glutamic oxaloacetic transaminase (SGOT). The normal ranges for

AST are laboratory specific, but can range from 3-45(units per litre)

(Allelo and Mays, 1998). Striking elevations of AST (400-4,000 units

per litres) are found in almost all forms of acute hepatic necrosis, such

as viral hepatitis and carbon tetrachloride poisoning (Parnigrahy et al.

1986). Usually low levels of AST are normally found in the blood.

When body tissues or an organ such as the heart or liver is diseased or

damaged, additional AST is released into the blood stream. The

amount of AST in the blood is directly related to the extent of the

tissue damaged.

Bilirubin is a lipid soluble substance usually carried in plasma as

a plasma protein bound substance. It is removed from the plasma

17

protein at the sinusoidal membrane of the liver cell, and it is thus

excreted into bile. It is a pigment derived from the decomposition of

hemoglobin within the red blood cell. It is also known as total

bilirubin, neonatal bilirubin and direct bilirubin. Fasting causes an

increase in the bilirubin level of the plasma, hence deep yellow

coloration of plasma is seen in a fasting animal (Aka, 2004).

Packed cell volume (PCV) is the variable normally used to

assess the basic status of the erythron-increased in polycythemia,

decreased in anemia although if a sample is too hemolyzed to allow

measurement of PCV, a meaningful Hemoglobin measurement may

still be obtained. An abnormally high PCV may be relatively due to a

change in the proportion of circulating RBC (Red Blood Cell) to blood

plasma without any alteration in the size of the erythron or absolute

due to a real increase in erythron size. The normal range of PCV for

poultry birds is between 30-40% (Allelo and Mays, 1998).

Hemoglobin (Hb) is a complex iron containing conjugated

protein. It is also composed of a pigment. Hemoglobin is contained in

the erythrocyte cell that gives it the characteristic red color. The

pigment contained in the hemoglobin is ferroheme. The protein

contained in hemoglobin is globin. Ferroheme makes up about 5% of

the hemoglobin. Thus the normal range is between 9-13g/dl (Allelo

and Mays, 1998).

18

CHAPTER THREE

MATERIALS AND METHODS

3:1 LOCATION AND DURATION OF STUDY.

The study was carried out at the Poultry Unit of Animal

Production Programme, Federal College of Agriculture Ishiagu,

Ebonyi State. The experiment lasted for Nine weeks (from February

2nd

to April 5th

2007). The first two weeks of the experiment period

served as adaptation and acclimatization period. The starter phase

lasted for three weeks, while the finisher phase lasted four weeks

respectively.

3.2 EXPERIMENTAL DESIGN

Ninety day old “ROSS” broiler chicks of mixed sexes from

“COSIN” Enugu were used for the study. The chicks were randomly

allocated to five treatment group with eighteen birds per treatment in a

completely Randomized Design experiment. Each treatment group

was further sub-divided into two replicate groups with nine birds per

replicate.

3:3 PROCESSING OF NEEM LEAF.

The Neem (Azadirachta indica) leaves used for the experiment

were obtained within the College environment. They were air-dried in

a ventilated room for one week. The leaves were thereafter sun dried

for about 2 hours at the environmental temperature of 30oC. They were

then reduced to smaller particles with pestle and mortar. Other feed

ingredients used were obtained from a Feed mill from Owerri in Imo

19

State and they include; maize (white maize), wheat offal,

soyabeanmeal, groundnut cake, palmkernelcake, fishmeal, bloodmeal,

bonemeal,vitamin-premix ,lysine and methionine while salt was

obtained from the local market.

3:4 EXPERIMENTAL DIETS

Five experimental diets were used in each phase of the study.

Treatment 1 ( Diet 1) served as the control with 0% of neem leaf meal,

while diets 2, 3, 4 and 5 contained neem leaf meal at 0.5, 1, 1.5 and

2% level of supplementation respectively ( Table 2).

20

Table 2:- Percentage Composition of Broiler Starter Diets

supplemented with Graded levels of Neem leaf meal.

Ingredients Diets

1 2 3 4 5

Neem leaf meal 0 0.5 1 1.5 2

Maize 44.50 44.40 44.20 44.10 44.10

Wheat offal 13.80 13.70 13.60 13.40 13.10

Soya bean meal 9.40 9.30 9.20 9.20 9.10

Groundnut cake 14.20 14.10 14.10 14.00 14.00

Palm kernel cake 7.50 7.40 7.30 7.20 7.10

Fish meal 5.70 5.70 5.70 5.70 5.70

Blood meal 1.90 1.90 1.90 1.90 1.90

Bone meal 2.00 2.00 2.00 2.00 2.00

Salt 0.25 0.25 0.25 0.25 0.25

Vitamin premix 0.25 0.25 0.25 0.25 0.25

Lysine 0.25 0.25 0.25 0.25 0.25

Methionine 0.25 0.25 0.25 0.25 0.25

Total 100 100 100 100 100

Calculated Analysis

Crude protein (%) 23.31 23.30 23.32 23.33 23.34

Energy (Kcal/k) 2,785.69 2,788.42 2,762.79 2,752.39 2,743.72

Crude fibre (%) 4.25 4.29 4.31 4.34 4.36

Determined Analysis Crude protein 24.06 24.28 23.62 24.53 24.65

Crude fibre 4 5 5.5 5.8 5.9

Ash 10 12 10 10 10

Ether extract 6 8 8 10 12

Moisture 10 10 12 10 12

NFE 45.94 40.72 40.88 39.67 37.45

21

Table 3:- Percentage Composition of Broiler Finisher Diets

supplemented with Graded levels of Neem leaf meal.

Ingredients Diets

1 2 3 4 5

Neem leaf meal 0 0.5 1 1.5 2

Maize 48.50 48.30 48.20 48.10 48.00

Wheat offal 14.10 14.00 13.90 13.80 13.70

Soya bean meal 5.40 5.30 5.20 5.10 5.00

Groundnut cake 12.30 12.20 12.10 12.10 12.00

Palm kernel cake 10.50 10.50 10.40 10.20 10.10

Fish meal 3.20 3.20 3.20 3.20 3.20

Blood meal 2.00 2.00 2.00 2.00 2.00

Bone meal 3.00 3.00 3.00 3.00 3.00

Salt 0.25 0.25 0.25 0.25 0.25

Vitamin-premix 0.25 0.25 0.25 0.25 0.25

Lysine 0.25 0.25 0.25 0.25 0.25

Methionine 0.25 0.25 0.25 0.25 0.25

Total 100 100 100 100 100

Calculated Analysis Crude protein (%) 20.17 20.16 20.15 20.16 20.15

Energy (Kcal/kg) 2,795.35 2,783.62 2,772.62 2,761.45 2,718.20

Crude fibre (%) 4.34 4.38 4.41 4.43 4.46

Determined Analysis Crude protein 20.45 20.57 20.56 20.71 20.60

Crude fibre 4.5 5.3 5.9 6.4 6.6

Ash 10 10 12 12 11

Ether extract 2.5 3.5 5 5 5

Moisture 10 10 10 12 12

NFE 52.55 50.63 46.54 43.89 44.80

22

3:5 MANAGEMENT OF BIRDS

Few days to the arrival of the chicks, the brooding pen, the

drinkers and the feeders were cleaned and disinfected. Wood shavings

were spread on the floor. The hovers, feeders and drinkers were placed

in position. Kerosene stoves and lanterns were used to provide the

heat needed to keep the temperature within optimum range (between

33oC to 35

oC). Feed and water were provided ad-libitum. Feeding was

carried out twice, in the morning between the hours of 7-8am and in

the evening between the hours of 5-6pm daily.

3:6 RESPONSE PARAMETERS

(a) Average Daily Feed Intake Per Bird:-

This was calculated as the amount of feed left over

subtracted from the total amount of feed supplied. The

result was then divided by the number of days of the

experiment (49 days).

(b) Average Daily Weight Gain Per Bird:-

This was estimated as the total weight of the birds at the

end of the period ( ninth week) less the total weight of the

birds at the beginning of the study, divided by the number

of days (49 days) and the result was then divided by the

number of birds per replicate.

(c) Feed Conversion Ratio:-

This was calculated as the average daily feed intake

divided by the average weight gain. 23

(d) Body Weight:-

This was measured on a weekly basis using a weighing

balance. The birds were however weighed at the beginning

of the experiment to determine their initial weight.

(e) Hematological Studies:-

This was carried out using blood samples collected from

each treatment. Two birds were collected from each

replicate (i.e. four birds per treatment) for analysis. The

parameters analyzed included; hemoglobin (Hb), packed

cell volume (PCV), total bilirubin and aspartate

aminotransferase (AST). They were determined according

to the methods of Okeudo et al. (2003)

3:7 STATISTICAL ANALYSIS

At the end of the study (nineth week) data obtained from the

response variables were subjected to analysis of variance using a one

way (ANOVA) by Steel and Torrie (1980) in a completely randomized

design (CRD)The experimental model of the CRD used in the analysis

is given as;

Yij = µ + Ti + Σij

Where;

Yij = Response variables of the observations on jth treatment.

µ = Mean of the common effect of the whole experiment.

Ti = Effect of the jth treatment.

Σij = Random error present in the observation and the treatment.

24

3:8 LABORATORY ANALYSIS

Some quantity of the dried neem leaf meal was obtained after

processing and taken for analysis in the laboratory at Michael Opara

University .Also some quantity of feeds formulated for the birds were

analyzed in the laboratory that is diet 1, 2, 3, 4 and 5 containing 0, 0.5,

1.0, 1.5 and 2.0% levels of inclusion of neem leaf meal respectively.

25

CHAPTER FOUR

RESULTS AND DISCUSSION

The result of the proximate composition of the Neem

(Azadirachta indica) leaf meal is presented in Table 6.

Table 6:- Proximate Composition of Neem (Azadirachta indica)

leaf meal.

Components Percentage

Crude protein 24.06

Ash 6

Moisture 3.5

Crude fibre 12

Ether extract 6

The result above showed that the Neem leaf meal contains,

24.06% crude protein, 6% ash, 3.5% moisture, 12% crudefibre and 6%

ether extract. This is similar to the result of the proximate composition

of the neem tree leave as presented by Esonu et al. (2006).

26

Table 7: Effects of Varying Levels of Supplemental Neem

(Azadirachta indica) leaf meal On the Performance

of Broiler Birds.

Dietary Treatment

Parameters Control (0%) 2(0.5%) 3(1%) 4(1.5%) 5(2%) SEM

Initial Body

Weight (g). 320 320 310 320 310 -

Avg. Final

Body Weight (g). 2320b 2520

a 2280

c 2270

c 1890

d 0.40

Avg. Daily

Wt. Gain (g). 40.82b 44.80

a 40.20

b 39.80

b 32.36

c 0.77

Avg. Daily

Feed Intake (g). 129.39a 131.71

a 128.24

a 126.56

b 110.60

c 0.85

Feed Conversion

Ratio (g). 3.17b

2.94c 3.19

b 3.18

b 3.43

a 0.05

4:1 Average Daily Weight Gain (g):-

Results presented in Table 7 shows that the highest daily

weight gain (g) was observed in birds fed diet 2 (0.5% NLM), while

the lowest was observed in birds fed diet 5 (2% NLM). It was

observed that the body weight gain of birds decreases with increase in

dietary levels of the NLM. The result agrees with the observation

made by D’Mello and Acamovic (1989), where graded dietary

inclusion of Leucaena leucocephala leaf meal induced cause related

depression in growth of chicken even when maize oil was used to

compensate for low metabolizable energy value of the leaf meal.

Similar observation of decreasing performance of broilers at high

27

levels of leaf meal inclusion have been observed by Nworgu and

Fapohunda (2002) who reported that the body weight gain of broilers

drastically reduced at high levels of mimosal leaf meal incorporation.

Also the work of Ranaweera et al. (1981) follows the same suit with

the incorporation of high levels of Gliricidia sepium leaf meal. The

effect observed above could have resulted from imbalance nutrient

value and improper metabolism associated with the neem leaf meal as

reported by Esonu et al. (2005). Other workers (Amaefule and Obioha,

2001 and Iheukwumere et al., 2002) have also reported the effect of

nutrient imbalance on monogastric animals fed high levels of

unconventional feed ingredients. This could also be probably due to

the effects of incomplete elimination of toxic factors as reported by

Udedibie et al. (1994) and Siddhuraju et al. (1996).

4:2 Average Daily Feed Intake:-

The analysis of variance conducted on the daily feed intake

shows that there was significant (P<0.05) differences among the

treatment means. The feed consumption of birds fed diet 1, 2 and 3

were similar (P>0.05) but significantly (P<0.05) higher than those of

birds fed diets 4 and 5. However the feed intake of birds fed diet 5 was

significantly (P<0.05) lower than that of birds fed diet 4. The observed

general trend was that of decreasing average feed intake as level of

supplementation increased. This might be attributed to the bitter nature

of the neem leaf which reduced the palatability of the feed. This is in

consonance with the observations of Bawa et al. (2006) who fed

rabbits with raw neem seed cakes and reported reduced feed

consumption among the rabbits on the test diets. Elangovan et al.

28

(2000) has also established the presence of bitter triterpenoids in the

neem seeds. However, debitterization through water washing, alkali

soaking and urea ammoniation to improve palatability has been

recommended (Katiyar et al., 1991). Also the decrease in feed intake

observed could be as a result of antinutritional factors present in the

test ingredient. This agrees with the findings of Nworgu, (2002) who

incorporated different leaf meals noted for their antinutritional factors

in the diets of poultry, and observed decreases in feed intake.

4:3 Feed Conversion Ratio:-

The analysis of variance shows that there were significant

(P<0.05) differences among the treatment means. As shown in table 7,

the amounts of feed consumed by birds fed diet 1(0% NLM), diet 3

(1% NLM) and diet 4 (1.5 % NLM) to gain the same unit (kg) of

bodyweight, were similar (P>0.05). However, birds fed diet 2 (0.5%,

NLM) were significantly (P<0.05) higher than those birds fed diet 5

(2%) to gain the same unit of body weight. The efficiency of feed

conversion of birds fed diets containing more than 0.5% NLM were

significantly (P<0.05) lower when compared to those of birds fed diet

2 (0.5%, NLM). The poor utilization of diets containing higher levels

of NLM might be related to the inability of the birds’ enzyme to break

down the active bitter substance in the NLM and also the imbalance

nutrient value and improper metabolism associated with the neem leaf

meal as reported by Esonu et al. (2005).

29

Table 8: Effect of Supplemental Neem Leaf Meal (Azadirachta

indica) On the Hematological Values of Broiler Birds

Dietary Treatments

Parameters 1 2 3 4 5 SEM

Hb (g/dl) 8.9b

10.8a 8.8

b 9.0

b 10.2

a 0.60

PCV (%) 32c 39

a 30

c 34

b 36

b 1.85

Total Bilirubin

(mg/dl) 2.35a 2.15

ab 2.25

a 2.15

ab 2.05

ab 0.21

AST (Unit/l) 10.05ab

10.15a 10.25

a 9.98

ab 10.10

a 0.24

4.5 Hemoglobin (Hb) Values:-

The result of the hematological values of broiler birds revealed

that there were significant difference (P<0.05) among the treatments.

Hb values of birds fed diets 2 and 5 were similar (P>0.05) but

significantly (P<0.05) higher than those of birds fed diets 1, 3 and 4,

which were themselves similar (P>0.05). The results of the

hemoglobin values of the birds on diet 3 and 1 fall slightly below the

recommended range of 9-13g/dl as reported by Allelo and Mays,

(1998). However, the values obtained for birds on diet 2, 4 and 5 fall

within the accepted range as reported by Allelo and Mays, (1998).

This result implies that including NLM in the diets of broilers as

supplemental feed had little or no effect on the relative quantity of the

complex iron containing the conjugate protein. This gives a clear-cut

reason why the redness of the broiler bird’s blood was not impaired.

30

4.6 Packed cell Volume (PCV) Values:-

The result of the Packed Cell Volume (PCV) revealed that birds

on diet 2, 4 and 5 were similar (P>0.05) but significantly (P<0.05)

higher than those of birds fed diet 1 and 3, which were themselves

similar (P>0.05). The highest value of packed cell volume was

obtained in birds on diet 2 (0.5% NLM). Thus the values obtained for

PCV across the treatment groups were within the normal range of 30-

40% as reported by Allelo and Mays, (1998). This result implies that

including NLM in the diets of broiler birds as supplemental feed

ingredient had little or no effect on the relative quantity of blood cells

as compared with the total volume of blood (Health and Olusanya,

1985).

4.7 Aspartate aminotrasferase (AST) Values:-

The result of the hematological values for Aspartate

aminotransferase (AST) shows that, there was no significant

difference (P>0.05) among the treatments. The result obtained for

AST across the treatment groups were within the accepted normal

range of 3-45 units/l as reported by Allelo and Mays (1998) and

Merk’s Veterinary Manual (1979). This is also in accordance to the

findings of Zinkl (1986) who reported that usually low levels of AST

are normally found in the blood of Poultry birds, but when high levels

are found (400-4,000unit/1) then there is likely to be cases such as

viral hepatitis and carbon tetrachloride poisoning.

31

4.8 Bilirubin Values:-

The result of the hematological values for total bilirubin shows that

there was no significant difference (P>0.05) among the treatments.

The low level of bilirubin obtained across the treatment groups is an

indication of the normalcy of the blood level of the birds. High levels

of bilirubin usually reveal that too much is being produced ( usually

due to increased destruction of red blood cells or that the liver is

incapable of adequately removing bilirubin in a timely manner ( due to

blockage of bile ducts and liver diseases such as cirrhosis and acute

hepatitis) (Aka, 2004).

32

CHAPTER FIVE

5.1 SUMMARY AND CONCLUSION

An experiment was conducted with Ninety “ROSS” unsexed day

old chicks to determine the optimum level of supplementation of neem

leaf meal in broiler diets. The experiment was laid-out in a completely

randomized design (CRD). The birds were randomly allocated to five

treatment groups of eighteen birds per treatment. Each treatment was

replicated twice with nine birds per replicate. The five groups were

randomly allocation to five diets containing varying levels of Neem

leaf meal (NLM) of 0% (diet 1), 0.5% (diet 2), 1.0% (diet 3), 1.5%

(diet 4) and 2.0% (diet 5). The bird’s performances were measured and

calculated on daily basis in terms of body weight gain, feed intake and

feed conversion ratio. At the end of the experiment the hematological

values were determined. The parameters determined for include

hemoglobin (Hg), aspartate aminotransferase (AST), bilirubin and

packed cell volume (PCV).

The results of this study showed that there were significant

(P<0.05) difference among the performance of birds across the various

treatments in terms of average body weight gain, feed intake, and feed

conversion ratio. Further supplementation of NLM beyond 0.5%

showed decrease in average bodyweight gain and average final weight

of the birds. This study also showed that the supplementation of NLM

had little or no effect on the hematological values of the birds, as the

parameters evaluated for across the treatment groups fell within the

accepted normal ranges for poultry birds.

33

The results of this study showed that the Neem leaf meal

supplementation in broiler diets could be effectively utilized at 0.5%

to obtain reasonably good growth rate and reduced disease infection

risk in the finished broiler birds.

5.2 SUGGESTION

It is strongly suggested that more research work be carried out or

undertaken with other species of poultry birds (like pullets, ducks,

geese, guinea fowl e.t.c) and other classes of monogastric and

ruminant animals as well. Also debitterization techniques as stated

earlier could improve feed intake when employed.

34

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41

APPENDIX. I

Average Daily Feed Intake

Treatments

Replicates 1 2 3 4 5

1 128.63 130.89 128.40 126.10 110.40

2 130.15 132.53 128.08 127.02 110.80

Sum 258.78 263.42 256.48 253.12 221.20

Mean 129.39 131.71 128.24 126.56 110.60

ANOVA TABLE

Sources of

variation

Degree of

freedom

Sum of

square

Mean

square

Fcal. Ftab.

Total sum

of square

9 571.3292

Treatment

sum of

square

4 568.2748 142.0687 232.56 5.19

Error sum

of square

5 3.0544 0.61088

42

APPENDIX. II

Average Daily weight gain

Treatments

Replicates 1 2 3 4 5

1 40.60 44.50 40.35 39.25 31.80

2 41.04 45.10 40.05 40.35 32.92

Sum 81.64 89.60 80.40 79.60 64.72

Mean 40.82 44.80 40.20 39.80 32.36

ANOVA TABLE

Sources of

variation

Degree of

freedom

Sum of

squares

Mean

square

Fcal. Ftab.

Total sum

of square

9 164.246

Treatment

sum of

square

4 162.692 40.673 130.866 5.19

Error sum

of square

5 1.554 0.3108

43

APPENDIX. III

Feed Conversion Ratio

Treatments

Replicates 1 2 3 4 5

1 3.17 2.94 3.18 3.21 3.48

2 3.17 2.94 3.20 3.15 3.37

Sum 6.34 5.88 6.38 6.36 6.85

Mean 3.17 2.94 3.19 3.18 3.43

ANOVA TABLE

Sources of

variation

Degree of

freedom

Sum of

square

Mean

square

Fcal. Ftab.

Total sum

of square

9 0.2437

Treatment

sum of

square

4 0.23565 0.0589 36.58 5.19

Error sum

of square

5 0.00805 0.00161

44

x