ANTIBODY RESPONSE OF FULANI ECOTYPE AND SASSO CHICKEN

INOCULATED WITH salmonella enteritidis

BY

AYOOLA OLADEHINDE AYOKULEYIN

MATRIC NO: 161884

A PROJECT SUBMITTED TO THE

DEPARTMENT OF ANIMAL PRODUCTION AND HEALTH,

FACULTY OF AGRICULTURAL SCIENCE, LADOKE AKINTOLA UNIVERSITY OF

TECHNOLOGY

IN PARTIAL FULFILMENT OF THE REQUIREMENT FOR THE AWARD OF

BATCHELOR OF TECHNOLOGY (B.TECH) IN ANIMAL PRODUCTION AND

HEALTH.

MAY 2023.
 DECLARATION
I hereby declare that this project was written by me and it is a record of my own research

work. All citations and sources of information are clearly acknowledged by means of references.

_______________________________ ______________________

AYOOLA Oladehinde A. Date

ii
 CERTIFICATION
This is to certify project titled “Antibody Response of Fulani Ecotype and Sasso

Chickens Inoculated with Salmonella enteritidis” by AYOOLA, OLADEHINDE

AYOKULEYIN with matric number 161884 meet the regulations governing award of the degree

of Bachelor of Technology, Ladoke Akintola University of Technology, Ogbomoso, Oyo State.

And is approved for it contribution to scientific knowledge and literary presentation.

_________________________________ ___________________
Prof. T.A Adedeji Date
B. Agric., M., Agric, Ph.D.
Professor of Breeding and Genetics
Supervisor

______________________
__________________________________ Date
Dr. T. A Rafiu
B. Agric., M., Agric, Ph.D.
Head of Department

iii
 DEDICATION
This project is mostly dedicated to God, the Alpha and Omega for his protection,

provision and guardian from the beginning of this project to the end. Also, I dedicate this project

to my mother and brothers for standing through for me.

1
 ACKNOWLEDGEMENTS
I would like to express my gratitude and appreciation to God Almighty for keeping my

life till the end of this project.

Also, I want to thank my project supervisor, Prof. T. A. Adedeji for the love he has

shown since the beginning till the end of this project, you have always been a role model and a

caring master and I want to say thank you for all.

I also want to say a very big thank you to the Head of Department, Animal Production

and Health, Dr. T. A. Rafiu and other staff of the department

My parents, especially my mother, I would say thank you for your prayers, word of

encouragement and trust and also thanks to my dad for making me a man.

To my brothers who has been supportive since the inception of this program, I

acknowledged you and very grateful for your love, trust and belief that you have in me.

Also, I must commend the effort of brother Oluwatobiloba Ojua and sister Abimbola for

the success of this project, also need to thank them for their teaching, assistance, word of

encouragement and advice.

To my group members, you are all wonderful, meeting you and being colleagues

throughout this project period is awesome. We had ups and downs but still remain a one big

bond. Thank you all. To my amiable friends, stay jiggy.

2
TABLE OF CONTENTS

Page

Title Page i

Declaration ii

Certification iii

Dedication iv

Acknowledgments v

Abstract

Table of Contents

List of Figures

CHAPTER ONE

1.0 Introduction 1

1.1 Justification 3

1.2 Objectives of the Study 4

1.2.1 General Objectives 4

1.2.2 Specific Objectives 4

CHAPTER TWO

2.0 Literature Review 5

2.1 Antibody Response 5

2.2 Salmonella 6

2.3 Salmonella enteritidis 7

2.4 Nigeria Indigenous Chickens Breed 8

2.5 Fulani Ecotype Chickens 9

3
2.6 The Exotic Breed Chickens 10

2.7 Sasso Chickens 11

CHAPTER THREE 12

3.0 Materials and Methods 12

3.1 Experimental Sites 12

3.2 Experimental Birds and Management 12

3.3 Experimental Feeds and Feeding 12

3.4 Experimental Mating 13

3.4.1 Mating Pattern 13

3.5 Egg Collection and Incubation 13

3.6 Candling and Hatching Process 14

3.7 Brooding, Chick Rearing and Management 14

3.8 Duration of the Experiment 15

3.9. Data Collection 15

3.10 Inoculation of Salmonella Disease Organism 15

3.11 Pre and Post Inoculation 15

3.12. Statistical Analysis 16

3.12.1. Antibody titre 16

CHAPTER FOUR

4.0 Results and Discussion 17

4.1 The bar chart of antibodies titre values of Nigerian indigenous and

exotic chickens inoculated with Salmonella enteritidis. 17

4.2 The bar chart of antibodies titre of Sasso Chickens inoculated

with Salmonella enteritidis. 17

4
4.3 The bar chart of antibodies titre of Fulani Ecotype Chickens both Male and female
with salmonella enteritidis. 18

4.4 Discussion 22

CHAPTER FIVE 24

5.0 Conclusion and Recommendation 24

5.1 Conclusion 24

5.2 Recommendation 24

References 25

5
 LIST OF FIGURES

Figure 4.1: The bar chart of antibodies titre values of Nigerian indigenous and exotic

chickens inoculated with Salmonella enteritidis

19

Figure 4.2: The bar chart of antibodies titre of Sasso Chickens inoculated with Salmonella

enteritidis.

20

Figure 4.3: The bar chart of antibodies titre of Fulani Ecotype Chickens with salmonella

enteritidis.

21

6
 ABSTRACT

This study was conducted to determine the antibody responses of Sasso and fulani ecotype

inoculated with salmonella enteritidis. The experiment was carried out at the at Teaching and

Research Farm, Lautech, Ogbomoso. 70 matured chickens of Fulani ecotype and Sasso were

sourced from the pre-existing stock on the farm and mating was done through artificial

insemination in a Completely Randomized Design. One hundred progenies were generated from

the crosses (Fulani ecotype × Fulani ecotype -20 chicks, Sasso × Sasso - 20 chicks) and raised

from day old. Cultured salmonella was sourced from Central Research Laboratory, ilorin, kwara

state. At 8 weeks old, the chicks were inoculated with 0.5ml of the cultured salmonella. Data

obtained was analyzed using SAS (2003) and significant means was separated using Duncan's

multiple range test. The results showed that there was significant (P 0.05) differences in the

laboratory titre of the chickens genotypes before and after inoculation with salmonella

enteritidis. The higher antibody titre before inoculation was recorded high (5) for Fulani ecotype

while Sasso chicken antibody titre was recorded medium (3) and after inoculation, antibody titre

of fulani ecotype was recorded low (2) while Sasso chickens was recorded low (2) respectively.

Therefore, it can be included that fulani ecotype and Sasso chickens are susceptible to salmonella

enteritidis but Nigeria fulani ecotype chickens are recommended as a good stock in terms of

disease resistance because it had the highest antibody titre before inoculation.

7
 CHAPTER ONE

1.0 INTRODUCTION

Salmonellosis in humans is a food borne bacterial disease which is mostly caused by

salmonella enterica mainly emanating from farm animals, food borne disease in humans caused

by salmonella continue to be a major public health concern worldwide and are commonly in

relation with the consumption of chicken meat contaminated with this bacterium (Rabsch et al.,

2010). Salmonella is a gram-negative minuscule, nonspore-forming, short rods (Popoff and Le

Minor, 2015). Salmonella enterica serovars enteritidis is one of the most frequently isolated

serovars from humans (WHO, 2007). Gastroenteritis is the most common manifestation of

salmonella infection worldwide, followed by bacteremia and enteric fever (Majowicz et al.,

2010).

The incidence of human infection and food poisoning by Salmonella has increased and

poultry are a major recognized source of infection (EFSA, 2007). Salmonella enterica infection

in chicken may occur at any age but it majorly affect chicks in their young age as the newly

hatched chicks are more susceptible to Salmonella enteritidis infection. Older birds builds

resistant to the infection due to the well developed immune system and strong cell mediated and

humoral immune responses (Beal et al., 2004).

Many serotypes of salmonella enteritidis has been identified with the ability to adapt within

various of animal hosts including humans, among some of the serotypes of salmonella that can

infect several animal species are salmonella typhimurium and salmonella enteritidis (Allerberger

et al., 2003). Salmonella enteritidis is able to outperform other salmonella antibody serotypes

with microbicidal elements by transferring lipular-like lipopolysaccharide (Huang et al., 2011).

8
 Nigerian indigenous chicken comprises of Normal feather, Frizzled feathers, Fulani

ecotype, Yoruba ecotype and Naked neck (Fayeye and Oketoyin, 2006). Indigenous chickens

have shown to be more disease resistant (Minga et al., 2004). The Nigeria indigenous chicken

has been used as a model organism for developmental and immunological studies and improving

animal health is a major goal in the current animal breeding industry (Burt and White, 2007).

Nigerian indigenous chicken are self reliant and hardy chicken with the capacity to withstand

harsh weather conditions and adaptation to adverse environment. They are known to posses

quality such as ability to hatch, brood and scavenge for major part of their food and possess

appreciated immunity from endemic diseases. Nigerian indigenous chicken needs to be

maintained for the purpose of conserving the wide disease resistance that they represent into the

future, they are of the highest value especially in this era of antibody resistance research and

enhance potential for the development of new improved breeds (Fulton, 2008).

The Fulani ecotype chicken is a native of Fulani tribe in the Middle belt and Northern parts

of Nigeria and are healthy and have better resistance to diseases and are adaptable to local

environment (Aro et al., 2013). Fulani ecotype chickens have survived various disease

challenges and environmental stress which has attested that the Fulani ecotype are naturally and

generally resistant to diseases like Salmonella enteritidis and others (Otim, 2005), resistance of a

Fulani ecotype chicken to a diseases is disease-specific.

Different exotic chickens are mostly introduced to be used in their pure or to distribute

them to the farming communities aimed at genetic improvement and building antibody resistance

to diseases and infections. There are different breeds of exotic chickens such as Rhode Island

Red, Australorp, New Hampshire, Sasso and White Leghorns (Demeke, (2008). Sasso chicken

have a low antibody resistance to Salmonella enteritidis and this may bring losses and also

9
spread rapidly through the flock and mortality maybe high (Tadelle and Ogle, 2001). Sasso

chicken is one of the exotic breed which originated from France and are know for some desirable

features such as feather color for camouflage, ability to escape from predators and disease

resistance (El-Safty, Ali and Fathi, 2006). Sasso being a newly introduced strain performs better

in responding to Salmonella enteritidis and other infections (Yakubu and Ari, 2017).

1.1 Justification

Antibody response is the defence of the host as a result of it's immune system to infections

or diseases, antibody levels are important dynamic parameters of immune response as they

partially reflect the potential of an animal to resist pathogen infection (Geng, 2007).

Improvement of disease resistance can be achieved through selection for increased immune

response (Bovenhuis et al., 2002). Improving antibody resistance to disease would reduce the

cost of vaccination and other disease prevention procedures and reduce mortality and loss in

performance during disease outbreak (Li et al., 2000). The emergence of Salmonella with

antimicrobial resistance is promoted by the use of antibiotics in animal feed to promote the

growth of food animals, and in veterinary medicine to treat bacterial infections in those animals

(Hyeon et al. 2011)

10
1.2 Objectives of the Study

1.2.1 General Objective

The general objective of the study is to determine the antibody response of Fulani ecotype

and Sasso chickens inoculated with Salmonella enteritidis

1.2.2 Specific Objectives

1. To determine the antibody response of Fulani ecotype chickens before and after inoculation

with Salmonella enteritidis.

2. To determine the antibody response of Sasso chickens before and after inoculation with

Salmonella enteritidis.

3. To access the antibody responses of fulani ecotype and Sasso chickens before and after

inoculation with Salmonella enteritidis.

11
 CHAPTER TWO

2.0 LITERATURE REVIEW

2.1 Antibody Response

The immune system is a naturally existing protective system for pathogen infection.

Vaccines stimulate specific immune responses to pathogens that provide animals with protection.

The development of effective vaccines against salmonella enterica serovar enteritidis for

chickens has been hindered by a lack of knowledge concerning the immune responses against

Salmonella in chickens, antibody responses, including humoral and cell-mediated responses,

play important roles in the resistance and clearance of Salmonella enteritidis infection, also,

there is sufficient evidence from various animal models that cell-mediated immunity plays a

major role in controlling salmonella infection (Lister, 2002) Antibodies can protect against

bacteria mainly by facilitating the uptake of the pathogen by phagocytic cells, which then destroy

the ingested bacteria (Attia, et al., 2012). Improvement of disease resistance might be achieved

through selection for increased immune response (Bovenhuis et al., 2002).

Innate immunity is the bird's first line of immune defence against a wide variety of

pathogens. Responses of the innate immune system are non-specific and do not distinguish

between invaders but respond to features that are common to many types of pathogens (Crump,

et al., 2008). The difference stages of Salmonella enteritidis infection are shown in a variety of

mechanisms of antibodies that contribute to the response against this bacterial infections

(Friedman, and Moon, 2008). Improving antibody resistance to disease would reduce the cost of

vaccination and other disease prevention procedures and reduce mortality and loss in

performance during disease outbreak (Li et al., 2000). Antibody response is the defence of the

host as a result of it's immune system to infections or diseases, antibody levels are important

12
dynamic parameters of immune response as they partially reflect the potential of an animal to

resist pathogen infection (Geng, 2007).

2.2 Salmonella

Poultry birds can become infected with many different types of salmonella; about 10

percent of all Salmonella species have been detected in poultry. The most important are

Salmonella typhimurium and Salmonella enteritidis. Infection may occur at any age in chicken

but the infection does occur during younger age as the newly hatched chicks are more

susceptible to salmonella infection, grown birds may build high resistant to salmonella enteritidis

infection as compared to the younger ones due to well developed immune system and strong cell

mediated and humoral immune responses (Beal et al., 2004). Poultry birds usually become

infected through contaminated feed, infected birds, eggs or environment. Salmonella is a gram-

negative minuscule, nonspore-forming, short rods (Popoff and Le Minor, 2015).

Prevention of bacterial infections especially those due to Salmonella in meat animals have

been of increased importance due to highly publicize food borne illnesses in humans related to

the consumption of the meat products (Fulton et al., 2002). Immunological methods detect

unique salmonella molecules using two antibodies; a surfacebound primary antibody to capture

the target molecule and a reporter antibody to detect the antibody target complex immunological

techniques (Beal et al., 2004 ), There is no vaccine available to prevent Salmonella infection.

However, one can take the following steps to help ensure good hygiene and food safety, reducing

the likelihood of getting the infection (Özoğlu and Altuntaş, 2019). Enzymes-linked

immunosorbent assay (ELISA) is attributed a rapid and highly sensitive test for detecting a

number of bird pathogens including salmonella infections (Ohore et al., 2002).

13
 There are two types of Salmonella including Salmonella enterica and Salmonella bongori,

Salmonella enteric is six species including salmonella typhi, Salmonella enteritidis, salmonella

paratyphi, salmonella typhimurium and Salmonella choleraesuis (Boyle et al., 2014). Salmonella

infection is one of the major health concern worldwide and it contribute to the keeping or raising

of animals with economic burden of industrialized and underdeveloped countries through the

costs associated with surveillance, prevention and treatment of disease (Crump et al. 2004).

2.3 Salmonella enteritidis

Salmonella enteritidis is a rodshaped, Gram-negative facultative anaerobe that belong to

the family enterobacteriaceae (Barlow and Hall, 2002), Salmonella Enteritidis had been used for

serological detection of infection in chickens (Holt et al., 2002), Salmonella enteritidis is a

zoonotic disease that of not properly taken care of could cause lose of lives in animals and

humans, the most common diseases are caused by suming chicken eggs containing the bacteria

Salmonella enteritidis (Velge et al., 2005). Salmonella enteritidis is one of the frequently isolated

serovars from animals (WHO, 2007). The observation of the availability of antibody in the

chickens infected with salmonella enteritidis was taken to see the effect of the antibody response

on the chickens after the inoculation of the infection (Gast, 2003).

There maybe wide spread of salmonella if the antibody response of the chicken is not

strong to withstand the challenge by a virulent field strain, the different components of the

immune response that protect against salmonella enteritidis infection in chickens is not fully

recovered and total host of immune response to bacterial invasion includes both humoral,

antibody-mediated and cellmediated immunity (Centres for Disease Control, (2000). The

antibody response of chickens to salmonella enteritidis determines how adaptive and receptive

chickens infected with salmonella can withstand and combat the infection (Holt et al., 2006)

14
 Salmonella enteritidis infection is one of the causes of death in chickens and this

disease can be transferred through chicken's products to humans. The use of antibiotics and

vaccines in the prevention and control of this infection are not totally efficient and also have side

effect, therefore, building the antibody resistance of chickens is a better approach in finding a

lasting solution

2.4 Nigeria Indigenous Chickens Breed

The indigenous chickens in Nigeria, which are mostly found in rural areas, are good

scavengers as well as foragers. They have good maternal qualities, high antibody, hardier when

compared to the exotic breeds and have high survival rates with minimal care and attention

(Salako and Ige, 2006). The frizzling and naked neck gene in particular have been described as

adaptability genes acting as disease resistant factor (Islam and Nishibori, 2009). There is high

immune response to salmonella enteritidis in normal feathered and frizzle feathered, Indigenous

chickens are the most commonly found across every corner of the tropical countries of Africa

where they are kept in the rural areas (Ajayi, 2010). Nigeria Indigenous chicken are self reliant

and hardy chicken with the capacity to withstand harsh weather conditions and adaptation to

adverse environment

Female indigenous chickens have a higher value than the male counterparts as a result of

high immune response in the animal, where females produce more vigorous humoral immune

reactions, which are more resistant to certain infectious diseases (Pitcovski and Cahaner et. al.,

2001). Report has it that frizzle and normal feather chicken genotypes are more resistance to

salmonella infection, Indigenous chicken needs to be maintained for the purpose of conserving

the wide disease resistance, they are of the highest value especially in this era of antibody

resistance research and enhance potential for the development of new improved breeds (Fulton,

15
2008). Antibody response to the same virus was different among chicken genotypes study,

Indigenous chicken have shown to be more disease resistance and more capable of utilizing low

quality feed (farell, 2000). Indigenous chickens have shown to be more disease resistant (Minga

et al., 2004). Nigeria indigenous chicken such as normally feathered chickens have high immune

response and disease resistance (Adriaansen-Tennekes et al., 2009).

2.5 Fulani Ecotype Chickens

The Fulani ecotype chicken is a native of Fulani tribe in the Middle belt and Northern

parts of Nigeria and are healthy and have better resistance to diseases and are adaptable to local

environment (Crump et al. 2004). Fulani ecotype chickens have survived various disease

challenges and environmental stress which has attested that the Fulani ecotype are naturally and

generally resistant to diseases like Salmonella enteritidis and others (Otim, 2005), resistance of

an Fulani ecotype chicken to a diseases is disease-specific. It is reported that chickens exhibit

different rate of immune response to different strain of the same pathogen (Msoffe et al., 2001).

A high lymphocytes count may be an indication of resistance to the Salmonella infection and

moreso, salmonella enteritidis is less invasive in the indigenous chickens such as Fulani ecotype

and others (Ishola and Holt 2008). Fulani ecotype chickens have been identified to have innate

antibody potentials that can be presented in commercial chicken production (Fayeye et al.,

2005). Some popular locally adapted chickens in Nigeria include the Fulani ecotype and Yoruba

ecotype (Ajayi, 2010), the Fulani ecotype build a high antibody resistance to diseases due to its

naturally raised environment, Fulani ecotype chickens are raised allowing chickens to freely

move where they would be exposed to alot or environmental challenges and are given less

vaccination due to the asymptomatic traits they have. (Centers for Disease Control and

Prevention, 2004).

16
 High prevalence of diseases is among the major factors limiting high productivity of the

local chickens in the tropics among these diseases is that caused by Salmonella enteritidis. Also,

Fulani ecotype developed response to thermal stress, drought, pathogens and suboptimal

nutrition (Olawunmi et al., 2008). The Fulani ecotype chicken has been described as a potential

meat type breed of chickens in Nigeria because of their body shape and weight (Fleming et al.,

2016).

The importance of imbuilt control in the resistance of infectious diseases is well recognized.

It has been suggested that some strains of Fulani ecotype chickens are more genetically

susceptible or resistant to infection with Salmonella than others (Berchieri et al., 2001). It has

been described that chicks are highly susceptible to Salmonella infection during the first 4 days

post hatch, becoming increasingly resistant to infection with age, such resistance requires cell-

mediated immunity (Mariani et al., 2001). Differences in resistance against Salmonella

enteritidis infections among different commercial lines (Mdegela et al., 2000) and in inbred

chicken lines have been reported (Berchieri et al., 2001).

2.6 The Exotic Breed Chickens

Exotic breed of chicken are susceptible to Salmonella enteritidis, the exotic breed of

chickens infected with salmonella enteritidis are more severe and last longer (Ishola and Holt,

2008). Different exotic chickens are mostly introduced to be used in their pure or to distribute

them to the farming communities aimed at genetic improvement and building antibody resistance

to diseases and infections. There are different breeds of exotic chickens such as Rhode Island

Red, Australorp, New Hampshire, Sasso and White Leghorns (Demeke, 2008). Sasso chicken

have a low antibody resistance to Salmonella enteritidis and this may bring losses and also

spread rapidly through the flock and mortality maybe high (Tadelle and Ogle, 2001). Sasso

17
chicken is one of the exotic breed of chickens that are not raised locally and are not exposed to

some environmental challenges to build high resistant to Salmonella enteritidis unlike the

indigenous breed of chickens

2.7 Sasso Chickens

Sasso chickens showed the best immune response to inoculation of salmonella enteritidis

and this could be attributed to the fact that the Sasso is a scavenging chicken genotype and

therefore could have similar immunological abilities, Sasso chickens are slow growing, robust,

easy to manage, multi-coloured birds which can be grown under different rearing systems from

traditional to intensive production system (Osei Amponsah et al., 2015). Sasso genotypes are

newly introduced into the Nigerian tropical environment; hence is dire need for their

characterization (Yakubu and Ari, 2016), it has been developed through an intensive selection of

traditional colored lines of chickens. Sasso breed is known for many desirable features of

indigenous birds, such as the feather colors for camouflage, ability to escape from predators,

resistance to diseases, adaptable to tropical and sub-tropical conditions. They have also the

ability to scavenge, thus require low maintenance yet grow about double the bodyweight of their

indigenous counterparts, provided that they receive supplementation and are protected against

diseases (Aman, Bangu, et al., 2017)

Despite the success recorded so far in the domestication of sasso chicken, disease attack

has been a challenging factor that has hampered poultry production in Nigeria. Salmonella and

Newcastle are two common diseases that affect poultry production (Sonaiya and Swan, 2004).

18
 CHAPTER THREE

3.0 MATERIALS AND METHODS

3.1 Experimental Sites

The experiment was carried out at the Poultry Unit of the Teaching and Research Farm,

Ladoke Akintola University of Technology, Ogbomoso (LAUTECH), Oyo State, Nigeria. The

site is located on longitude 4 0 15‫ ׳‬East and latitude 80 15‫ ׳‬North east of the Greenwich meridian.

The altitude is between 300 and 600m above sea level. The mean annual rainfall and temperature

are 1247 mm and 270 respectively (BATC, 2006). Ogbomoso is located in a derived savannah

zone. The other part of the experienment was carried out at Animal Production and Health

Department Laboratory, Ladoke Akintola University of Technology, Ogbomoso.

3.2 Experimental Birds and Management

The experimental birds consisted of Fulani ecotype and Sasso chickens. The parent

chickens was sourced from pre-existing flock at Teaching and Research Farm, at maturity (19-20

weeks). Total of ten cocks (10) and sixty (60) hens was used as parent stocks and distributed a 2

cocks to 10 hens in each genotype. Birds was individually wing tagged for identification

purposes. Cocks and hens was caged in an open-sided house of 1.161m 2. Each bird was confined

in a cell space of 15 × 15. Prior to stocking, the whole pens and cages was washed and

disinfected using Lysol® and was left for five days before stocking.

3.3 Experimental Feeds and Feeding

The cocks were fed with commercial grower mash containing 15.0% Crude Protein and

2500kcal Metabolizable Energy while the hens were fed with commercial layer mash containing

16.0% Crude Protein and 2800kcal Metabolizable Energy. Clean water was supplied.

19
3.4 Experimental Mating

Mating of the birds was done using artificial insemination (AI) at 24 weeks old in the farm.

Hand massage technique involving application of gentle slight and pressure at the back

(massage) of the cocks towards the tail was adopted in training the cocks for semen production.

This training commenced at 20 weeks old. The massaging was done twice a day for 4 weeks.

Afterwards, feather around the vent was shaved and repeated at two weeks interval.

Semen was collected by pushing the cocks tail upward with left hand and at the same time

using the thumb and the fore finger of the right hand to press the copulatory organ and semen

was discharged into sterilized semen-collection containers. Collection was diligently done to

avoid contamination from faecal droppings and blood.

3.4.1 Mating Pattern

Fulani Ecotype (sire) × Fulani Ecotype (dam) (FE)

Sasso (sire) × Sasso (dam) (SA)

3.5 Egg Collection and Incubation

Total number of 50 eggs was collected from each of the Fulani ecotype and Sasso

chickens. Fertile eggs was collected every day and arranged in the crates with the pointed ends

down and pedigreed along sire and dam genotype lines. Eggs was allowed to accrue and stored at

room temperature of 320c . In the hatchery, the eggs was set in a cabinet type incubator at a

commercial hatchery in Ibadan, Oyo State, Nigeria. The eggs was set along sire and dam

genotype lines at a temperature between 27 - 390c and relative humidity of 55 – 56% for eighteen

days. The temperature was increased to 29 - 40 0c at a relative humidity of 70 – 75% afterwards,

from nineteenth day to hatching time. The eggs was turned automatically through 90 0c in the

incubator.

20
3.6 Candling and Hatching Process

Candling of eggs was carried out on the 19th day of incubation for the identification of

fertile and infertile eggs. The process was carried out in a dark room using a Candler fixed with

neon fluorescent tubes. The eggs was set on the Candler for easy penetration of light through the

eggs and each egg was viewed against the source of light. After candling, the fertile eggs was

transferred into the hatching tray according to sire and dam lines and taken to the hatcher. After

hatching, the day old chicks was left in the hatcher until 90% dried.

3.7 Brooding, Chick Rearing and Management

Before the arrival of the day old chicks (DOC), the brooder pens was washed with

Lysol® and again disinfected three days prior to arrival. The dimension of each brooder pen was

8 × 10m2 and tagged along the sire and dam genotype lines. Wood-shaving was spread on the

floor of each pen and heat source was placed at a corner in the brooder pen to provide warmth

for the chicks. The heat was supplied for 4 weeks.

The chicks was brooded for four weeks. Adequate heat, ventilation, medication and

feeding was provided. The chicks was duly vaccinated and antibiotics drugs was administered as

required. The two genotypes was reared together in deep litter pens but all the experimental birds

was wing tagged for proper identification.

At the arrival of DOC, they were given multi vitamin drugs which served as an anti-

stress. Commercial chick mash containing 22.0% Crude Protein and 2900Kcal Metabolizable

Energy was fed to the chicks from day old till eight weeks of age. Commercial grower mash

containing 15.0% Crude Protein and 2500Kcal Metabolizable Energy was introduced to the

chicks gradually from eight weeks till when the birds were twelve weeks. Clean water was

supplied ad-libitum to the birds. The litter was changed at 3days intervals to prevent

21
accumulation of ammonia gas. Spillage of water on the litter was prevented as much as possible.

All chicks remained in deep-litter throughout the period of the experiment.

3.8 Duration of the Experiment

The experiment was conducted for six months.

3.9. Data Collection

Data was collected on antibody titre of the chicken used.

3.10 Inoculation of Salmonella Disease Organism

Salmonella enteritidis disease vaccine was purchased at laboratory in Ilorin Kwara State.

The experiment birds was inoculated at 8 weeks of age with Salmonella disease organism. This

was done subcutaneously at 0.5ml (LD50) per birds as recommended by the manufacturer. The

process was separated after 5 weeks of the inoculation in order to ensure that all the birds is

properly challenged with Salmonella disease.

3.11 Pre and Post Inoculation

Blood samples was collected twice on chicks pre and post-inoculation, before and after

inoculation. 2ml of blood was collected from each experimental birds using steroids needle and

syringe. One mililitre (1ml) from the blood sample collection was transferred into a well labelled

plain bottle and was allowed to coagulate to obtain the serum for slide agglutination test (using

widal test indicator serum) to determine the level of antibiotics against Salmonella. This was

compared with the base-line result obtained at 7th week before inoculation. The remaining 1ml

blood sample was transferred into EDTA bottle for hermatological analyses, using routinely

available clinical methods as described above.

22
3.12 Statistical Analysis

Data collected on antibody titre was subjected to General Linear Model of SAS (2003).

Data collected reproductive traits was subjected to one-way analysis for the effect of genotype

(SAS 2003). Significant means were separated by Duncan's Multiple Range Test. The statistical

models are as shown.

3.12 .1 Antibody titre

The values of antibody titre against Salmonella was transferred using logarithmic
X+1
transformation (Log10 ) and thereafter was analyses using General Linear Model (GLM) of

SAS of 9.0 software 2003. Means with significantly difference are separated by Duncan's

Multiple range test of the same software. The model adopted is as specified below:

Yij = µ + Bi + Sj + (B × S)ij + eij

Where;

Yij = response variable

µ = Overall mean,

Bi = fixed effect of the breed (i = 1, 2),

Sj = fixed effect of jth sex (j = 1, 2),

Bij×Sij = fixed effect of the interaction between breed and sex of bird,

eij = experimental error which is evenly distributed.

23
 CHAPTER FOUR

4.0 RESULTS AND DISCUSSION

The bar chart 1 to 4 presented the graphical representation of the antibodies titre on the

scale of 1-2 (low), 3-4 (medium) and 5-6 (high) antibodies titre before and after inoculation. The

bar chart revealed that there is significant effect of genotype and sex on the antibodies titre of the

chicken genotypes studied before and after inoculation with salmonella enteritidis.

4. 1 The bar chart of antibodies titre values of Nigerian indigenous and exotic chickens

inoculated with Salmonella enteritidis.

Figure 4.3 revealed the antibody titre of Sasso and Fulani ecotype chickens before and

after inoculation with Salmonella enteritidis. Before the inoculation with Salmonella enteritidis,

the antibody titre of Fulani ecotype chickens was high (5) while that of Sasso chickens was

medium (3). Antibody after inoculation fall to low (2) for both breeds which shows that there

was no significant effect of genotype on the antibody titre of all chickens genotype studied.

4.2 The bar chart of antibodies titre of Sasso Chickens inoculated with Salmonella

enteritidis

Figure 4,2 presented that the antibody titre of Sasso chickens before inoculation with

Salmonella enteritidis was on medium level (3) which after inoculation of Salmonella enteritis

reduced to low level (2) . Therefore, there was significant effect on the antibody titre of the

chicken genotypes studied.

4.1 The bar chart of antibodies titre of Fulani Ecotype Chickens inoculated with

salmonella enteritidis

24
Figure 4.1 showed that the antibody titre of Fulani ecotype before inoculation with Salmonella

enteritidis was recorded high (5) and which after inoculation of Salmonella enteritidis, the

antibody titre was recorded low (2). Therefore, there was significant effect on the antibody titre

of the chicken genotypes studied.

25
 6

3
SA
FE
2

0
Antibody before inoculation Antibody after inoculation

Figure 4.1: The bar chart of antibodies titre values of Nigerian indigenous and exotic chickens

inoculated with Salmonella enteritidis. SA = Sasso Chicken, FE = Fulani Ecotype Chicken.

26
 6

3 SA
Series2
2

0
Antibody before Antibody after Male Female
inoculation inoculation

Figure 4.2: The bar chart of antibodies titre of Sasso Chickens before and after inoculation with

Salmonella enteritidis.

27
 FE
6

4
FE
3

0
Antibody before Antibody after Male Female
inoculation inoculation

Figure 4.3: The bar chart of antibodies titre of Fulani Ecotype Chickens before and after

inoculation with salmonella enteritidis.

28
4.4 Discussion

The presence of some levels of 1gG antibody by week 1 post infection especially in the

group infected with Salmonella enteritidis agrees with the previous report where presence of

serum antibodies by most birds that were subcutaneously infected with salmonella enteritidis by

one week post infection had been observed (Gast, 2003). The poor Salmonella enteritidis

antibody response of birds challenge could probably be due to lack of establishment of infection

as a result of inadequate challenge dose or quick elimination of the organism from the system by

the host defense mechanism this stimulating fewer antibody producing cells. Therefore, chickens

exposed to few Salmonella colonies during artificial infection may not show evidence of

infection and continue to spread the organism within the birds. The implication of this finding is

that if inadequate vaccine dose is given to chickens during vaccination, they may respond with

sub-optimal immune response which may not be strong enough to withstand challenge by a

virulent field strain.

The Fulani ecotype chickens of Nigeria indigenous chickens have good adaptive

performances to hot and humid tropics, they have good maternal qualities, high antibody, hardier

and have high survival rate with minimal care and attention (Salako and Ige, 2006). Female

Fulani ecotype indigenous chickens have a higher value of resistance than the male counterpart

as a result of high immune response in animal where females produce more vigorous humoral

reactions which ate more resistant to certain infectious diseases like salmonella enteritidis and

salmonella typhimurium (Pitcovski and Cahaner et al., 2001). Sasso chickens are not highly

resistance to Salmonella enteritidis due to it breeding genesis. Although, Fulani ecotype chickens

are the best in keeping or raising for commercial purpose because of it antibody titre value

towards virulent infections.

29
30
 CHAPTER FIVE

5.0 CONCLUSION AND RECOMMENDATION

5.1 Conclusion

From the results of the study, Fulani ecotype chicken which is a Nigerian indigenous

chicken had the highest antibodies titre before and after inoculation.

5.2 Recommendation

The Nigerian indigenous chickens are recommended as a good stock in terms of disease

resistance.

31
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