EFFECT OF DIETARY TOASTED LEUCAENA LEUCOCEPHALA SEED MEAL ON GROWTH AND FEED UTILIZATION OF CLARIID CATFISH HETEROCLARIAS (Clarias gariepinus ♀ X Heterobranchus longifilis ♂)

POST FINGERLINGS

BY

Dave Efosa OMOSIGHOAGR1000192

DEPARTMENT OF FISHERIESFACULTY OF AGRICULTURE

UNIVERSITY OF BENINBENIN CITY

OCTOBER, 2015

EFFECT OF DIETARY TOASTED LEUCAENA LEUCOCEPHALA SEED MEAL ON GROWTH AND FEED UTILIZATION OF CLARIID CATFISH HETEROCLARIAS

i

(Clarias gariepinus♀ X Heterobranchus longifilis ♂) POST FINGERLINGS

BY

Dave Efosa OMOSIGHO

AGR1000192

A PROJECT WORK SUBMITTED TO THE DEPARTMENT OF FISHERIES FACULTY OF AGRICULTURE,

UNIVERSITY OF BENIN, BENIN CITY IN PARTIAL FULFILLMENT OF THE REQUIREMENT FOR THE

AWARD OF BACHELOR OF AGRICULTURE DECREE, B. AGRIC (FISHERIES)

OCTOBER, 2015

CERTIFICATION

ii

This is to certify that this project work was carried out by Dave Efosa OMOSIGHO

(AGR1000192) of Department of Fisheries, Faculty of Agriculture, University of Benin,

Benin City. In fulfilment for the award of Bachelor of Agriculture (BSc. Agric) degree in

Fisheries of University of Benin.

_______________ ______________

Dr. B. S. Aliu Date

Project Supervisor

_________________ _______________

Dr (MRS) F. A. R. Ehigiator Date

Head of Department

DECICATIONiii

Unto Him that is able to do exceeding abundantly above all that I can ever ask or think, to

the One who is, was, and is to come, the Lord God Almighty, the Lord Strong and Mighty,

my Abba Father I dedicate this report for His grace and endless love.

To my late father, Mr S. O. Omosigho Ogie, who passed-on on the 6th February, 2014.

ACKNOLEGDEMENTS

iv

First of all, gratitude goes to my maker, king, Lord, father, best friend, companion, solace,

gem, my confidant and inspiration, lover of my soul, ‘THE LORD GOD ALMIGHTY’ you

alone all glory be ascribed. And to my wonderful, supportive and inspirational project

supervisor Dr. B. S. Aliu for his fatherly love, care, direction and encouragement.

My profound gratitude goes to my late father, Mr S. O. Omosigho Ogie, mother – Mrs

Patience Omosigho and siblings – Mr Nosa, Mr Lucky, Mr Clement, Mrs gladdys, Mrs

Loveth, Mrs Abieyuwa, Miss Adesuwa, Mrs Esohe, Mrs Imuetiyan, Mr Johnbull, Mr

Rapheal, Mr. Ivie, and Mrs. Esther for their love, care, prayers, encouragement,

constructive criticism and financial support all through these years.

With utmost gratitude, I want to specially appreciate the lecturers in the Department of

Fisheries for their parental care, moral and academic investments into my life – Dr (MRS)

F. A. R. Ehigiator (HOD), Prof. F. A. Oguzie, Prof O. J. Abolagba, Prof. V. A. Okonji, Dr

B. S. Aliu, (project supervisor), Dr (MRS) Odiko (course adviser), Dr S. Wangboje, Mr.

marinus, Mr. Kenneth, Mrs Glory and Miss Ihenyen may God richly reward your labour of

love, to the farm officers I would not forget in a hurry – Mr Iriowen E. Nosakhare, Miss

Aniekan Dickson, God richly bless you and to the Lab. Technician Miss. Eseosa, God bless

you ma.

Also my appreciation goes to my project mates Josephine, Victor and Perpetual.

Finally to my friends Nsikanabasi, Tony, Joseph, Cresentia, Praise, Evezi, Festus, Clifford,

Francis, Greg, Habibat, Stanley, Ivy, Jesse, Shaka, Peace, Victory, CJ, Meshack, Agas,

Precious, Benjamin, Solomon, Samojo, Levi, Ejaeta and Ailele who have made my

schooling fun all the way, I love you all. To my room-mates and course mates whom I

cannot mention one and leave the others. May God bless you all.

v

TABLES OF CONTENTS

Title page …………………………………………………………………………………….i

Certification …………………………………………………………………………...…...iii

Dedication………………………………………………………………………………......iv

Acknowledgements …………………………………………………………………………v

Table of contents …………………………………………………………………………..vi

List of tables……………………………………………………………………………….viii

One figure…………………………………………………………………………………..ix

List of plates ………………………………………………………………………………..x

Abstract ……………………………………………………………………………………xi

CHAPTER ONE

1.0 Introduction …………………………………………………………………………1

1.1 Justification of the Study ……………………………………………………………4

1.2 Objectives of the Study………………………………………………………………5

CHAPTER TWO

2.0 Literature review……………………………………………………………………6

2.1 Fish Nutrition ………………………………………………………………………7

2.2 Protein and amino acids requirement of heteroclarias …………………………….8

2.3 Carbohydrate/energy requirement of heteroclarias ………………………………11

2.4 Lipid and fatty acids requirement of heteroclarias ……………………………….12

2.5 Vitamin requirement of heteroclarias …………………………………………….13

2.6 Mineral requirement of heteroclarias ……………………………………………..14

2.7 Fish feed ingredients ………………………………………………………………15

2.8 Leucaena leucocephala …………………………………………………………....17

2.9 Chemical composition and nutrient profile of leucaena leucocephala seed……….20

vi

CHAPTER THREE

3.0 Material and Method ………………………………………………………………23

3.1 Preparation of Leucaena and Soybean meal ………………………………………23

3.2 Preparation of Experimental Diets …………………………………………………23

3.3 Experimental Fish …………………………………………………………………25

3.4 Experimental Units ………………………………………………………………...25

3.5 Experimental Procedure …………………………………………………………...25

3.6 Parameters Monitored …………………………………………………………….26

3.7 Proximate Analysis of Diets and Fish …………………………………………….27

3.8 Statistical Analysis ………………………………………………………………...27

CHAPTER FOUR

Results ……………………………………………………………………………………28

CHAPTER FIVE

Discussion …………………………………………………………………………………34

CHAPTER SIX

Conclusion and Recommendations ………………………………………………………..37

References …………………………………………………………………………………38

vii

LIST OF TABLES

Table 1: Protein requirement of some commonly cultured fish species at

different life stages in Nigeria …………………………………………...…………10

Table 2: Chemical composition of Leucaena leucocephala seeds …………………………20

Table 3: Amino acid composition of Leucaena leucocephala seeds ………………………22

Table 4: Percentage composition of experimental diets ……………………………………24

Table 5: Proximate Composition (%) Of Experimental Diets …………………………….29

Table 6: Carcass composition (%) of Heteroclarias post fingerlings

fed varying levels of L. leucocephala seed meal based diets for 56 days …………30

Table 7: Growth response and nutrient utilization of Heteroclarias post

fingerling fed Leucaena leucocephala seed meal based diets …………………...…31

viii

ONE FIGURE

Fig. 1: Chemical Structure of Mimosine …………………………………..………………21

ix

LIST OF PLATES

Plate 1: Leucaena leucocephala tree…..…………………………………………………...18

Plate 2: flower of Leucaena leucocephala …………………………………………………19

Plate 3: Leucaena leucocephala seed ………………………………………………………19

x

ABSTRACT

An experiment was designed and carried out to assess the growth performance and feed

utilization (weight gain, feed conversion ratio, protein efficiency ratio, specific growth rate,

feed intake and survival) of Heteroclarias post fingerlings (48.66g) fed graded levels of

toasted Leucaena leucocephala seed meal based diets with the aim of establishing the best

inclusion level of Leucaena seed meal. One hundred and ten (110) post fingerlings with an

initial mean weight of 48.66g were allotted at random to six treatments in triplicate groups

with each treatment tank having five post fingerlings and were fed with isocaloric and

isonitrogenous diets containing 40% crude protein (CP). The toasted seed meal was used to

replace soybean meal in the diets in the following proportions: diet I (0%), diet II (10%),

diet III (20%), diet IV (30%), diet V (40%) and diet VI (50%). At the end of the feeding

trials that lasted for 8 weeks, the mean weight gain of fish, feed intake and specific growth

rate showed no significant difference (P > 0.05) between treatment I and II, but were

significantly different from treatments III, IV and V. The percentage weight gain and

protein efficiency ratio were not significant difference (P > 0.05) among all the treatment

while feed conversion ratio was significant difference (P > 0.05) among all the treatment.

Although, the mean weight gain of fish, percentage weight gain, feed intake and feed

conversion ratio was highest in treatment I, followed by treatment II in mean weight gain

of fish, percentage weight gain, feed intake and feed conversion ratio and highest in protein

efficiency ratio and specific growth rate, while, mean weight gain of fish, Feed intake,

specific growth rate, feed conversion ratio and protein efficiency was lowest in treatment xi

IV. Percentage weight gain was lowest in treatment VI. Survival among treatments slightly

varied significantly but was not as a result of the feed consumed. At the end of the

experiment, the recommended inclusion level of Leucaena seed meal was 10%.

xii

CHAPTER ONE

1.0 INTRODUCTION

Aquaculture the farming of aquatic organisms including fish, molluscs, crustaceans and

aquatic plants is necessary to meet the protein need of Nigerians. Over time, there has

been increase in fish production in Nigeria. Aquaculture of fish is one of the fastest

growing food producing sectors in the world accounting for approximately 50% of

fisheries products (Food and Agriculture Organisation (FAO), 2010). According to

Bello (2007) and FAO (2005), the artisanal fish production level increased by 5.4%,

aquaculture fish production by 4.3% and industrial fishery through the use of trawlers

by 12% over the previous years. However, of this increase in production, the desired

result has not been attained. Quantitatively, details of fish production as at 2005 stood at

490,600tons from the artisanal fisheries, 56,300 tons from industrial fishery through the

use of trawlers; while fish importation stood at 61,150 tons. In meeting up with the

growing need for fish production, aquaculture practise has been identified as a possible

alternative; the reasons being that the activities of artisanal and industrial fishery in our

natural waters have led to over exploitation and degradation due to human activities in

our coastal water. To fully bring aquaculture to its desired level, four production

challenges have been identified, these are: feeding the fish stock in the pond with high

quality fish feed, management of good pond water quality, provision of high quality

fish seeds and pond construction. The first two challenges: fish feeding and water

quality management directing affect each other. The level of feeding of the stocks

affects the water quality and level of water quality affects the feeding performance of

fish in the pond (George, 2001). Nutrition which is the combination of processes by

which a living organism receives and utilizes the materials necessary for the

maintenance of its function and for the growth and renewal of its components (Olomu,

2011). Fish nutrition deals with nutrient requirement of fish and its availability, the best

fish feed produces the highest production with limited environmental impact (Omitoyin,

2007). According to Craig and Helfrich (2002) and Houlihan et al., (2001), nutrition is

critical in fish farming even as it represents 40-50% of total fish production costs. This

further explains that the standard of fish farming and levels of production largely

depends on the standard of fish nutrition. Although fish nutrition has advanced

dramatically in recent years with the development of new and balanced commercial

1

diets that promote optimal fish growth and health, protein which averages between 18-

42% depending on the fish breed (Craig and Helfrich, 2002) remains the basic

requirement in formulated fish feeds for fish growth and production. The use of

imported fish feed by fish farmers is affected by its high cost therefore making it

unaffordable to small scale fish farmers, particularly in the developing countries

(Olusegun and Eniade, 2014). Fish feed which is compounded mainly using fishmeal as

the protein source is made from whole bodies of small bony fish species generally

considered not suitable for direct human consumption. This is the highest quality

protein source commonly available to fish feed manufacturers. Thus fishmeal protein is

being used globally as dietary protein in formulated fish feeds as accounted by Hardy

and Tacon (2002); New and Wijkstom (2002); Krishnankutty (2005); Yigit et al.

(2006). The continued expansion of aquaculture will not be possible if fishmeal is

relied upon as the main source of protein in aquacultural feeds (Nogueira et al., 2012)

which is due to its relatively high and variable cost. Therefore, it is desirable to replace

fishmeal with less expensive protein sources, this has necessitated researches into the

use of plants protein in feed formulation such as soybean, oilseed, groundnut cake, etc.

Among the plant protein sources considered in aquaculture diets, soybean meal is the

most widely used ingredient. It was used for the replacement of fish meal at various

rations due to their high-protein content and relatively well-balanced amino acid profile,

(Koumi et al., 2009). However, due to the fact that soybean is a conventional protein

feed stuff utilised by man, livestock and fish, its demand has increased, supply reduced,

and cost prohibitively increased, thus, there is need for the use of non-convectional

plant protein sources (Obe, 2014). Studies have shown that non-convectional vegetable

protein sources (legumes) in particular have high potentials for supplying protein need

for maximum productivity after been properly processed (Pillay, 1990; Nwanna et al.,

2008). The need for such recommendations have been due to the presence of certain

limiting factors in those ingredients such as high crude fibre content (Nwanna et al.,

2008), antinutritional factors (Alegbeleye et al., 2001).

Leucaena leucocephala, a non-convectional plant protein, is generally known as

“multipurpose tree” (Jones, 1997) due to its diverse use. L. leucocephala was reported

capable of producing about 3-5 tonnes seeds ha-1 yr-1 (D’Mello, 2000) and has a crude

protein (CP) value of 28 to 45% (Widin, 2004; Atawodi et. al., 2008). It is known to be

high in α-carotene (Kale, 1987) with a rich amino acid profile. L. leucocephala 2

demonstrated good potential as a useful plant protein source in fish ration (Ter Muelen

et al., 1981; D’Mello and Acamovic, 1989).

Heteroclarias is an intergeneric hybrid of two African Clariid catfishes;

Heterobranchus longifilis (male) and Clarias gariepinus (female) (Adewumi, 2014).

Clarias gariepinus occupies a unique and prominent position in commercial fisheries in

Nigeria because of its ability to be tasty, hardy as well as tolerant to poor water

conditions, it has an efficient feed conversion ratio in the males and as such attracts

high market price, reaches maturity early and has higher fecundity than

Heterobranchus longifilis (Nwadukwe, 2000). It is also capable of reproducing in

captivity and growing to a size of about 7.0 kg (Idodo Umeh, 2003; Nweke and

Ogwumba, 2005; Ekelemu, 2010). In addition, Similarly, Heterobranchus longifilis can

tolerate low dissolved oxygen, high turbidity and importantly grows faster to a size of

14.0 kg, has higher feed conversion but not as hardy and does not have the same high

survival rate as Clarias gariepinus (Vincent et al., 2014). Several studies have

demonstrated that Heteroclarias (Clarias gariepinus ♀ X Heterobranchus longifilis ♂)

exhibit superior growth, improved survival and general hardiness than true breed of

either Clarias gariepinus or Heterobranchus longifilis (Madu et al., 1991; Salami et al.,

1993; Nwadukwe, 1995).

This study therefore examined the nutrient potentials of processed Leucaena Seed

Meals (LSM) in the diet of Heteroclarias.

1.1 JUSTIFICATION OF STUDY

Feed constitutes 60-70% of total investment in aquaculture. The high cost of fish feed

has been recognized as a major factor militating against rapid development of

aquaculture in the developing countries due to most of the conventional feedstuffs being

used in human foods and animal feeds hence bringing about exorbitant prices and

scarcity of these feedstuffs.

There is the need to production of low cost fish feeds which is due to high cost,

declining supply and increased level of competition. In a bid to produce feed at reduced

cost and lessen the pressure on conventional feedstuff, studies are been carried out on

the use of unconventional feedstuffs as alternative source of protein for different fish

species. Soybean is an example of such feedstuff, commonly used as a protein source

3

in fish diets. There is therefore a necessity for research into non-conventional protein

ingredients that can replace soybean without compromising fish growth and health. This

study is aimed at determining the effect of dietary toasted Leuceana seed meals in the

growth and utilization of Heteroclarias.

1.2 OBJECTIVES OF THE STUDY

The objectives of this study are as follows:

1. To determine the effect of toasted Luecaena luecocephala on the growth of

Hetroclarias post fingerlings

2. To determine the utilization of Luecaena luecocephala by Hetroclarias post

fingerlings.

3. To determine the survival rate of Hetroclarias post fingerlings fed with

Luecaena luecocephala seed meal.

4. To determine the optimum inclusion level of Luecaena luecocephala seed

meal for overall performance of Hetroclarias post fingerlings.

4

CHAPTER TWO

2.0 LITERATURE REVIEW

Fish farming involves raising fish in tanks or enclosures, usually for food or

commercial purposes. Fish farming has been proven to be a very important sector in the

Nigerian economy and a successful way of enhancing fish production in the world

(International Food Policy Research Institute, (IFPRI) 2003). Fish farming in Nigeria is

currently a very lucrative business and it is mainly boosted by the continuous rise in the

demand for the African catfish. This trend therefore makes catfish culture the most

popular form of fish farming in Nigeria with Clarias gariepinus, Heteroclarias and

Heterobranchus spp. being the most desirable for culture (Adekoya et al.,2006), and

they have remained an important species for research in aquaculture.

Fish and fisheries is an important source of food, income, employment, and recreation

for people in the world and it is a very important source of animal protein for both man

and livestock in developed and developing countries (Davies and Davies, 2009;

Emmanuel et al., 2014). In Nigeria, fish and fish products constitute more than 60% of

the total protein intake in adults especially in rural areas (Adekoya, 2004). According to

Fagbenro and Adeparusi (2003), Fish contains protein of very high quality and has

sufficient amounts of all the essential amino acids required by the body for maintenance

of lean tissues which makes it a very important food for humans. Amiengheme (2005)

enumerated the importance of fish in human nutrition as follows: food fish having a

nutrient profile superior to all terrestrial meats (beef, pork and chicken) is an excellent

source of high quality animal protein and high digestible energy; fish is a good source

of sulphur and essential amino acid such as lysine, leucine, valine and arginine. It is

therefore suitable for supplementing diets of high carbohydrate contents; fish is also a

good source of thiamine as well as an extremely rich source of Omega-3-polysaturated

fatty acid, fat soluble vitamin (A, D and E) and water soluble vitamins (B-complex) and

minerals (calcium, phosphorus, iron, iodine and selenium); it has a high content of

Omega-3-polysaturated fatty acid, which are important in lowering blood cholesterol

level and high blood pressure. It is able to mitigate and to alleviate platelet of

(cholesterol) aggregation and various arteriosclerosis conditions in adult populations; it

reduces the risk of sudden death from heart attacks and reduces rheumatoid arthritis; 5

Omega-3 fatty acids also lower the risk of age related muscular degradation and vision

impairment; and it decreases the risk of bowel cancer and reduces insulin resistance in

skeletal muscles.

2.1 FISH NUTRITION

Nutrition is the combination of processes by which a living organism receives and

utilizes the materials necessary for the maintenance of its function and for the growth

and renewal of its components (Olomu, 2011). Good nutrition in animal production

system is essential to economically produce a healthy, high quality product. In the wild,

fish search for food and they are able to meet their body needs and as such they rarely

show signs of nutritional deficiency (Oresegun, 2004). When fish are taken from the

wild to an artificial and confined environment, the right amount of food should and

must be supplied so as to enable them grow and carry out other required activities (Eyo,

2003). In fish farming, nutrition is critical because feed represents 40 – 50% of the

production cost (Houlihan et al., 2001). Fish requires high quality nutritionally balanced

diet for growth and attainment of market size within the shortest possible time (Gabriel,

2007). Fish nutrition deals with nutrient requirement of fish and its availability, the best

fish feed produces the highest production with limited environmental impact (Omitoyin,

2007). Nutrition aims at providing all essential nutrients in adequate amount and in

optimum proportions (Oso et al, 2006). Fish like other animals have a requirement for

essential nutrients in order to grow properly. Increased understanding of the nutritional

requirement for various fish species and technological advances in feed manufacturing

have allowed the development and use of manufactured/artificial diets (formulated

feeds) to supplement or replace natural feeds in the aquaculture industry (Winfree,

1992). It is not surprising therefore that fish nutrition has become one of the most

important research and development components within aquaculture development

today. The development of a semi-intensive or intensive feeding regime for fish

requires first a basic understanding of the nutrition and dietary nutrient requirements of

the animal. With the exception of water and energy, the dietary nutrient requirements of

all aquaculture species can be considered under five different nutrient groups; proteins,

lipids, carbohydrates, vitamins, and minerals.

2.2 PROTEIN AND AMINO ACIDS REQUIREMENT OF

HETEROCLARIAS

6

Proteins are very complex organic compounds of high molecular weight. In common

with carbohydrates and lipids, they contain carbon (C), hydrogen (H), and oxygen (O),

but in addition also contain about 16 % nitrogen (N: range 12–19%), and sometimes

phosphorus (P) and sulphur (S). Proteins are among the most important constituents of

all living cells and represent the largest chemical group in the animal body, with the

exception of water; the whole fish carcass contains on average 75% water, 16% protein,

6% lipid, and 3% ash. Proteins are essential components of both the cell nucleus and

cell protoplasm, and accordingly account for about 70 percent of the dry weight of fish

muscle (Robinson et al., 2006), internal organs, brain, nerves and skin. It is therefore an

essential nutrient for both maintenance and growth in fish (Falayi, 2009). A continual

supply of protein is needed throughout life of the fish. Protein is a major constituent and

it is the most expensive part of fish feed (Fagbenro et al., 1992), generally, increasing

protein levels in fish diets can lead to improved fish production though it may be

expensive (Diyaware et al., 2009). Thus, it is important to accurately determine the

protein requirements for each species and size of cultured fish (Craig and Helfrich,

2002). Fish utilizes both plant and animal proteins although animal proteins have

superior nutritional qualities (such as crude protein content and amino acid profile but

they tend to be more expensive (Eyo, 2003). Proteins are formed by linkages of

individual amino acids. Although over 200 amino acids occur in nature about 23 amino

acids have been isolated from natural proteins and only about 20 amino acids are

common. Of these, ten are essential (indispensable) amino acids that cannot be

synthesized by fish. The ten essential amino acids that must be supplied by the diet are:

methionine, arginine, threonine, tryptophan, histidine, isoleucine, lysine, leucine, valine

and phenylalanine. Of these, lysine and methionine are often the first limiting amino

acids. A dispensable amino acid is one that can be synthesized by the animal in

quantities sufficient for maximum growth (Jackson et al., 2003).The first need when it

comes to protein requirements of fish is to supply the indispensable amino acid

requirement of the animal and secondly to supply the dispensable amino acids or

sufficient amino nitrogen to enable their synthesis (Macarthney, 1996).

Protein requirements generally are higher for smaller fish (see Table 1). As fish grow

larger, their protein requirements usually decrease. Protein requirements also vary with

rearing environment, water temperature and water quality, as well as the genetic

composition and feeding rates of the fish. Protein is used for fish growth if adequate 7

levels of fats and carbohydrates are present in the diet. If not, protein may be used for

energy and life support rather than growth. Dietary protein requirements of fish have

been reported by several authors: Falayi (2009), a range of 25-30% crude protein for

Tilapia and Carp, Jamabo and Ockiya (2008), reported 40% crude protein for

Heterobranchus. bidorsalis, while Fagbenro et al. (1992), reported 42.5% dietary

protein requirement for Heterobranchus longifilis, Diyaware et al. (2009), reported

50% crude protein for catfish hybrid of Heterobranchus bidorsalis (♂) x Clarias

anguillaris (♀) under laboratory conditions.

Table 1: Protein requirement of some commonly cultured fish species at different

life stages in Nigeria

Fish Protein Stage of development

Protein(%C.P)requirement

H. bidorsalis Fry/fingerlings 45-60

Juveniles 40-50

Adult/Broodstock 35-45

C. gariepinus Fry/fingerlings 45-60

Juveniles 40-50

Adult/Broodstock 35-45

O.niloticus Fry/fingerlings 35

Juveniles 28-30

Adult/Broodstock 22-30

Source: Akintomide et al. (2008)

8

2.3 CARBOHYDRATE/ENERGY REQUIREMENT OF HETEROCLARIAS

Carbohydrates are compounds of carbon, hydrogen and oxygen. They usually occur in

three different forms which are sugar, starch and fiber. Carbohydrates are the most

economical and inexpensive sources of energy for fish diets. Fish like most animals eat

to meet their energy requirements. Carbohydrate is one of the most important parts of

fish diet (Robinson et al., 2006). Although not essential, carbohydrates are included in

fish diets to reduce feed costs and for their binding activity during feed manufacturing.

Cooking starch during the extrusion process makes it more biologically available to fish

(Craig and Helfrich, 2002). The mechanism of utilization in fish varies between

carnivorous and omnivorous fish (Falayi, 2009). Catfishes cannot efficiently utilize

carbohydrate as only about 1.6 kcal can be extracted from same amount of carbohydrate

(Ayoola, 2011). Up to about 25-30% of dietary carbohydrate can be used by fish (Toft,

2001).

In fish, carbohydrates are stored as glycogen that can be mobilized to satisfy energy

demands. They are a major energy source for mammals, but are not used efficiently by

fish. Fish can produce energy from lipids and protein making carbohydrate a non-

essential constituent in their diet for normal growth and functions. Robinson et al.,

(2006), stated that even though catfish do not need carbohydrates in their diet, catfish

feed contains considerable carbohydrates supplied from grains and grain by-products

(such as corn grain, wheat grain and wheat middlings) that are rich in starch. The ability

of catfish to utilize starch as an energy source gives rise to the protein sparing effect of

carbohydrate (Eyo, 2003). A typical catfish feed contains 25 percent or more soluble

(digestible) carbohydrate plus 3-6 percent more carbohydrate that are usually present as

crude fibre (mainly cellulose).

The energy requirement for catfish has it has been reported by Robinson et al., (2006),

to be generally expressed as a ratio of digestible energy (DE) to crude protein and it

ranges from 7.4 to 12 kilocalorie/gram (kcal/g). An increase in the ratio of digestible

energy to crude protein above this range may bring about increased fat deposition,

reduced processed yield and shorter shelf life and if the value is too low, the fish will

grow slowly.

9

2.4 LIPID AND FATTY ACIDS REQUIREMENT OF HETEROCLARIAS

Lipids (fats and oils) are a highly digestible source of concentrated energy. It contains

about 2.25 times as much energy as does an equivalent amount of carbohydrates

(Robinson et al., 2001). Lipids play several important roles in an animal’s metabolism,

such as supplying essential fatty acids (EFA), serving as a vehicle for absorption of fat-

soluble vitamins, and serving as precursors for steroid hormones and other compounds

(Robinson et al., 2006).

Simple lipids include fatty acids and triacylglycerol. Fish typically require fatty acids of

the omega 3 and 6 (n-3 and n-6) families. Fatty acids can be: a) saturated fatty acids

(SFA, no double bonds), b) polyunsaturated fatty acids (PUFA, >2 double bonds), or c)

highly unsaturated fatty acids (HUFA, > 4 double bonds). Marine fish oils are naturally

high (>30%) in omega 3 HUFA, and are excellent sources of lipids for the manufacture

of fish diets in quantities ranging from 0.5 – 2.0% of dry diet (Steven and Louis, 2002).

The type and amount of lipid used in catfish diets is based on essential fatty acid

requirements, economics, constraints of feed manufacture, and quality of fish flesh

desired (Ibiyo and Olowosegun, 2005). A recent trend in fish feeds is to use higher

levels of lipids in the diet in order to spare protein partially and to cut costs of diets

although excessive fat deposition in the liver can decrease the health and market quality

of the fishes (Francis-Floyd and Millie, 2012). African catfish has the ability to

synthesize most of their fatty acids; thus, nutritionally there may be no “best” level of

dietary lipid except that needed to provide EFA. Catfish apparently require 0.5 percent

to 0.75 percent omega-3 fatty acids in diet (Robinson et al., 2006).

2.5 VITAMIN REQUIREMENT OF HETEROCLARIAS

Vitamins are a heterogeneous group of organic compounds essential for the growth,

reproduction, health and maintenance of animal life including fish. The variety and

amount of vitamins are so complex that they are usually prepared synthetically and are

available commercially as a balanced and premeasured mixture known as a vitamin

premix (additives) (Akintomide et al., 2008; Abowei and Ekubo, 2011). The vitamins

that are present in feed ingredients are usually not considered during feed formulation

because their bio availability is not known, but they certainly contribute to the amount

10

of vitamin present in the feed (Edwin and Meng, 1996). The majority of vitamins are

not synthesized by the fish body or at a rate sufficient to meet the fish needs and

therefore must be supplied in the diet (Halver, 2002). Vitamin premix is added to the

diet of fish in generous amounts to ensure that adequate levels of vitamins and minerals

are supplied to meet dietary requirements (Shapawi et al., 2007).

The two groups of vitamins are water-soluble and fat-soluble. Water-soluble vitamins

include: the B vitamins, choline, inositol, folic acid, pantothenic acid, biotin and

ascorbic acid (vitamin C). Of these, vitamin C probably is the most important because it

is a powerful antioxidant and helps the immune system in fish (Craig and Helfrich,

2002). The fat-soluble vitamins include A vitamins, retinols (responsible for vision); the

D vitamins, cholecalciferol (bone integrity); E vitamins, the tocopherols (antioxidants);

and K vitamins such as menadione (blood clotting, skin integrity). Of these, vitamin E

receives the most attention for its important role as an antioxidant. According to Ayo

(2011), all animals including fish, display distinct morphological and physiological

deficiency signs when individual vitamins are absent from their diet. Deficiency of each

vitamin has certain specific symptoms, but reduced growth is the most common

symptom of any vitamin deficiency, Scoliosis (bent backbone symptom) and dark

coloration may result from deficiencies of ascorbic acid and folic acid vitamins,

respectively. (Craig and Helfrich, 2002). According to Cruz-Suarez et al. (2007),

Ascorbic acid added to feeds should be phosphorylated to stabilize the vitamin and

increase storage time. Generally, Catfish feeds are supplemented with a vitamin premix

that contains all essential vitamins in sufficient quantities to meet the requirement and

to compensate for losses due to feed processing and storage (Robinson et al., 2006).

Shapawi et al. (2007), reported that 60 mg/kg of vitamin C is recommended for

catfishes.

2.6 MINERAL REQUIREMENT OF HETEROCLARIAS

Minerals are defined as organic compounds animals require in small amounts in their

diet for normal growth, health and reproduction. They are inorganic elements necessary

in the diet for normal body functions (Akintomide et al., 2008). Minerals can be divided

into two groups (macro-minerals and micro-minerals) based on the quantity required in

the diet and the amount present in fish (Steven and Louis, 2002). Macro-minerals

include: Calcium (Ca), sodium (Na), chloride (Cl), potassium (K), Phosphorus (P),

11

Magnesium (Mg) and Sulphur (S). Micro-minerals (trace minerals) are required in

small amounts as components in enzyme and hormone systems. Micro-minerals

include: copper (Cu), zinc (Zn), selenium (Se), chromium (Cr), iodine (I), Manganese

(Mn) iron (Fe), cobalt (Co), fluorine (F), vanadium (Va), tin (Sn), silicon (Si) and

molybdenum (Mo). According to O’Keefe and Newman (2011), the functions of

minerals in fish are as follows:

they are structural components of hard tissues,

they are components of soft tissues,

they are cofactors and activators of enzymes,

they function in acid-base balance,

they function in the production of membrane potentials,

they are essential for the transmission of nerve impulses and muscle contraction

they play a key role in osmoregulation.

Catfish apparently require the same minerals for metabolism and skeletal structure as

other animals require. Although these mineral are required in small amounts as

compared to proteins, lipids and carbohydrate, they are critically important and the

deficiency of one or more of these micronutrients or its excess addition in the diet can

be detrimental to fish health. Minerals are potentially lethal when present in amounts

slightly above or below the requirement and accurate supplementation are therefore

imperative for proper diet formulation (National Research Council (NRC), 1993).

Amongst the minerals that are required by catfish, phosphorus is particularly important

in fish feeds because it is required by fish in a fairly large amount (Nwanna et al., 2008)

Feedstuffs, especially those from plants, are poor sources of phosphorus and fish do not

get enough phosphorus from pond water. Due to this fact, catfish feed are usually

supplemented with phosphorus. African catfish feeds are typically supplemented with

mineral premix with enough of all the essential minerals to meet or exceed dietary

requirements of the fish (Robinson et al., 2006).

2.7 FISH FEED INGREDIENTS

12

Nutrients essential to fish are the same as those required by most other animals. These

include water, proteins (amino acids), lipids (fats, oils, fatty acids), carbohydrates

(sugars, starch), vitamins and minerals, thus a nutritious diet is required for proper

growth and high protein quality for better market value (Abowei and Ekubo, 2011).

Feedstuffs are classified as conventional or unconventional base on its acceptability and

usage in fish feed formulation. Some conventional feedstuffs are groundnut cake,

soybean meal, palm kernel meal, brewers dried yeast, brewers dried grain, maize, wheat

offal, and fish meal. The quality and proportion in which these conventional feedstuffs

are used depends on its nutrient composition, presence of anti-nutritional substances,

palatability and cost (Robinson et al., 1998).

Unconventional feedstuffs are potential feed ingredients, which have hitherto not been

used in fish feed production for some certain reasons like;

they are not well known or understood,

no effective study of the method of production with a view to commercializing

them,

they are not readily available, and

they can be toxic or poisonous (Abowei and Ekubo, 2011).

They contain high quality feed ingredients that can compare favourably with

conventional feed types. They are usually cheaper by the virtue of the fact that there is

no competition for human consumption (Roberts, 1989). Unconventional feedstuffs can

be of animal or pant source. Some of these unconventional feedstuffs of animal origin

include tadpole meal, fly larvae, earthworm meal, toad meal, shrimp waste, crab meal,

poultry-hatchery waste meal and animal waste such as pig and poultry droppings and

blood meal. Plants sources include leaf protein, leaf meal, aquatic macrophytes,

cultivable pulses such as mucuna bean, yam beans, bread beans and winged beans

(Abowei and Ekubo, 2011). Fish feed can be compounded by the farmer using a

combination of available ingredients that are found within their locality (Omitoyin,

2005). No single feed ingredient can supply all of the nutrients and energy required for

optimum growth of catfish, thus, commercial catfish feeds contain a mixture of

feedstuffs and vitamin and mineral premixes that provide adequate amounts of essential

nutrients, as well as the energy necessary for their utilization (Robinson et al., 2001).

13

The amount of each feed ingredient used depends on several factors including nutrient

requirements, ingredient cost, availability of each ingredient, and processing

characteristics.

2.8 LEUCAENA LEUCOCEPHALA

Leucaena leucocephala is a medium sized fast growing tree belongs to the family

Fabaceae (see plate 1). It is native to Southern Mexico and Northern Central America

and now it has naturalized in many tropical and sub-tropical locations (Chandrasekhara

et al., 1984). The specific name ‘leucocephala’ comes from ‘leu’ meaning white and

‘cephala’, meaning head, referring to the flowers. It is commonly known as White Lead

tree, White Popinac, Jumbay and Wild Tamarind. In India, it is popularly known as

kubabul or subabul. During the 1970s and 1980s it was promoted as a "miracle tree"

due to its multiple uses (Suttee, 2002). It has also been described as a "conflict tree"

because it has been promoted for its forage production and naturally spreads like a

weed. It grows up to 20m height. Leaves are looking like that of tamarind having white

flowers tinged with yellow (see plate 2), and having long flattened pods. Seeds are dark

brown with hard shining seed coat (see plate 3) (Meena et al., 2013). The tree has

multifarious uses such as like firewood, timber, greens, fodder, green manure, provide

shade, controls soil erosion (Shelton and Brewbaker, 1994). The kernel of seeds

contains more than 20% oil and it can be used as a bio energy crop. The seeds may also

be used as concentrates for dairy animals, as manure, as a protein source, as an oil seed

and as a potential source of commercial gum (Gardezi et al., 2004). The plant has high

nutritive contents like protein, carbohydrates and fat as that of alfalfa.

Leucaena leucocephala is one of the numerous legume grains and shrubs that are in

abundance in Nigeria playing a very important role in ecological and biodiversity

conservation as well as in ruminant farming in the country (Sotolou and Faturoti, 2008).

The common type is short, bushy, up to 5m in height and flowers when very young,

about 4 - 6 months (Suttee, 2002). According to D’Mello (2000) and Sotolu and

Faturoti (2008), when conditions are favourable, it can flower all year round and this

brings about the production of abundant seeds (about 3-5 tonnes seeds per hectare per

year).

14

Plate 1: Leucaena leucocephala tree (source: North Coast Weeds Advisory Committee

(NCWAC), 2015)

15

Plate 2: flower of Leucaena leucocephala (source: www.uncommoncactus.com)

Plate 3: Leucaena leucocephala seed (source: www.ornatejewels.wordpress.com)

16

2.9 CHEMICAL COMPOSITION AND NUTRIENT PROFILE OF

LEUCAENA LEUCOCEPHALA SEEDS

Leucaena seeds are oval in shape and have brown hulls and yellow kernels (Aderibigbe

et al., 2011). Leucaena leucocephala leaves and seeds contain lipids, crude protein,

carbohydrates, fibre, calcium, phosphorus, and tannin (see Table 2).

Table 2: Chemical composition of Leucaena leucocephala seeds

Item Composition

ME (Metabolizable energy) 2573.26kcal/kg

Crude protein 311.00g/kg

Crude fat 56.00g/kg

Crude fibre 132.00g/kg

Dry matter 948.00g/kg

Crude ash 45.00g/kg

NFE 404.00g/kg

Calcium 3.70g/kg

Total phosphorus 3.40g/kg

Tannin 0.75

Phytate mg/100g 697.50

Source: Mohamed et al. (2009)

The hull: kernel ratio is 50:50 by weight (Sethi and Kulkarni, 1995). The seeds are low

in oil, 5.1 - 10% and rich in protein 24.5 - 46% (Sotolu and Faturoti, 2008). The

kernels have an oil content of 11.9 - 15.3% and a protein content of 52.5 - 66.4%. Thus,

the nutrients are concentrated in the kernels (Sethi and Kulkarni, 1995). It is significant

that the proteins of L. leucocephala seeds are fairly rich in the essential amino acids

(see Table 3) isoleucine, leucine, phenylalanine, histidine, lysine and methionine

(Mohammed et al., 2009). According to Kale (1987), the seeds are known to be high in 17

α carotene. Of the fatty oils contained in the seeds, approximately 26 - 29% is saturated

acids and 71 - 73% unsaturated acids. The oil is rich in linoleic acid (42.5 - 65%).

Leucaena seed has high calcium and phosphorus levels (Sethi and Kulkarni, 1995).

The genus Leucaena is also reported to contain hydrocyanic acid, leucaenine, quercitrin

and tannic acid (James, 1983). The use of L. Leucocephala seeds limited by the

presence of mimosine (see fig. 1), a non-protein amino acid substance (NRC 1984)

which constitute up to 60% (2,689.5mg) of the total free amino acids (4,885.8mg) in the

seeds (Sethi and Kulkarni, 1995).

Fig. 1: Chemical Structure of Mimosine (Source: www.bio.miami.edu/mimosa

/Mimosine.gif)

The mimosine content of the Leucaena plant vary considerably in different species

(Kewalramani et al., 1987) and even within the same species, that is, L. leucocephala

itself, for the various cultivars. The presence of anti-nutritional factors (ANFs) such as

mimosine are the main factors responsible for their limited use if feed production. It is

thus recommended that vegetable meals are to be properly processed before it is used in

the practical fish feeding (Francis et al., 2001). Heat treatment methods are commonly

the most efficient and often used methods of detoxifying anti-nutrients that are present

in grain legumes and other vegetable matters. The heating method could either be wet

(boiling) or dry (toasting or autoclaving). Other method include soaking and subsequent

sun drying of the seeds.

Table 3: Amino acid composition of Leucaena leucocephala seeds18

Amino acids g/kg g/16Gn

Cysteine 3.5 1.13

Arginine 26.20 8.42

Methionine 3.60 1.16

Glutamic acid 46.30 14.89

Threonine 8.70 2.80

Glycine 13.80 4.44

Alanine 11.10 3.57

Valine 11.10 3.57

Isoleucine 9.30 2.99

Leucine 18.10 5.82

Lysine 13.90 4.47

Methionine +cysteine 7.10 2.28

Source: Mohamed et al. (2009)

CHAPTER THREE

19

3.0 MATERIALS AND METHODS

3.1 PREPARATION OF LEUCAENA AND SOYABEAN MEAL

Leucaena seeds where harvested from the L. leococephala shrubs that were found

around the Faculty of Agriculture, University of Benin. Matured and dried pods

(brown) were collected and placed into plastic containers. The seeds were later sundried

for two days to ensure all seeds were dried uniformly.

A pot was placed on fire and heated up with low heat for 2 minutes before the seeds

were introduced into the pot. They were stirred continuously for 10-15 minutes or till

the seed coat became reddish brown in colour and emitting a sweet aroma. They are

then remove and allowed to cool before milling to flour. It was there-after sieved to

remove the skin of the seeds. Fresh soyabean seed where purchased from Uselu market

in Benin City and poured into a preheated pot and toasted in a similar manner as was

carried out with the Leucaena seeds and milled.

3.2 PREPARATION OF EXPERIMENTAL DIETS

Fishmeal, corn meal, wheat offal and bone meal, multivitamin capsule (vitamin premix)

and vitamin E-gel that were used in the production of the feed were purchased from a

private company at Murtala Mohammed Way in Benin City. The multivitamin capsule

(vitamin premix) and vitamin E-gel were purchased from a pharmaceutical shop and the

palm oil was obtained from the market in Benin City.

Six isonitrogenous and isocaloric diets with a crude protein level of 40% were

formulated. Diets 1 (control), 2, 3, 4, 5, 6 had soybean meal protein substituted with

leucaena seed meal at 0%, 10%, 20%, 30%, 40%, 50% respectively. The composition

of the experimental diets is shown in Table 4.

Table 4: Percentage composition of experimental diets20

TREATMENTS

INGREDIENTS I II III IV V VI

% substitution leucaena seed

meal

0% 10% 20% 30% 40% 50%

Fishmeal (65.5% CP)

Soyabean (38.8% CP)

Leucaena seed meal (40%

CP)

Maize (9.5% CP)

Wheat offal (13% CP)

Palm oil

Bone meal

Vitamin premix

Vitamin E (gel)

Total

25.00

54.00

0.00

3.00

4.00

8.00

4.00

1.00

1.00

100.00

25.00

44.00

10.00

3.00

4.00

8.00

4.00

1.00

1.00

100.00

25.00

34.00

20.00

3.00

4.00

8.00

4.00

1.00

1.00

100.00

25.00

24.00

30.00

3.00

4.00

8.00

4.00

1.00

1.00

100.00

25.00

14.00

40.00

3.00

4.00

8.00

4.00

1.00

1.00

100.00

25.00

4.00

50.00

3.00

4.00

8.00

4.00

1.00

1.00

100.00

(Source: As analyzed by me)

The required quantity of ingredient for each of the diet were weighed and mixed into a

homogenous mixture with the exception of corn meal. The component of corn meal in

each diet was gelatinized (boiled in water to form gel), which served as a binder was

poured on the homogenized mixture and mixed properly. They were all made into

pellets with a pelleting machine available in the Department of Fisheries experimental

farm at the University of Benin. The diets were dried using Altona smoking kiln in the

department of fisheries experimental farm and then stored in air-tight containers

throughout the experimental period.

3.3 EXPERIMENTAL FISH21

One hundred and ten (110) heteroclarias post-fingerings were obtained from the

hatchery unit of the Department of Fisheries experimental farm, University of Benin.

They were acclimatized for 72hrs in the laboratory during which they will be fed

commercial feeds.

3.4 EXPERIMENTAL UNITS

Eighteen rectangular (18) plastic tanks (six (6) treatment in three (3) replicates)

measuring (30cm×36cm×52cm) were used. The experiment was carried out at the

Departmental of Fisheries wet laboratory, University if Benin. Each tank was filled with

water up to 2/3 of its volume with bore-hole water attached to the laboratory.

3.5 EXPERIMENTAL PROCEDURE

At the end of acclimatization the fishes will be weighed in batches of 5 into each of the

experimental units replicated three for each treatment. They were fed twice daily to

satiation to ensure maximum growth between 8:00 - 9:00hrs and 15:00 - 16:00hrs.

Feeding was monitored for each unit to ensure that fishes were not underfed or overfed.

The experimental units were cleaned daily by siphoning with a thin hose to remove

unconsumed feed and faecal waste of the fish to reduce pollution of the water. About

two-third of the water will be drained and replaced during cleaning before feeding.

Total replacement of water was made during the weekly weighing of fish. All fish per

treatment were weighed and counted weekly to determine growth and survival.

3.6 PARAMETERS MONITORED

Data on feed consumed and weight gain were collected weekly for each unit from

which the following performance parameters were evaluated.

1. Weight gain (WG) = W2 – W1 (g)

Where; W1 = initial weight

W2 = final weight

2. Feed intake = initial weight of feed – final weight of feed

22

3. Specific growth rate (SGR) % = Loge W 2−logeW 1

T 2−T 1X 100

Where: T1 and T2 are time of experiment in days.

W2 = final weight at T2

W1 = initial weight at T1

Loge = natural logarithm.

4. Percentage weight gain (PWG) % = Weight Gain

Initial WeightX 100

5. Food conversion ratio (FCR) % = Feed Intake(g)

Wet Weight Gain(g)X 100

6. Protein efficiency ratio (PER) % = Weight Gain(g)Protein Intake

X 100

7. Survival rate % = Initial stocked−mortality

Initial stockedX 100

3.7 PROXIMATE ANALYSIS OF DIETS AND FISH

A sample of 12 fishes of the initial stock as well as some survivals from each treatment

was sacrificed at the commencement and end of the experiment. Diet samples from the

six diets were also collected. They were analysed using standard methods of the

Association of Official Analytical Chemists (AOAC) (1990) for protein, fat, ash and

moisture. The moisture content was determined by heating the samples in an oven at a

temperature of 105°C for 24 hours and recording the weight loss, the crude protein

content was estimated by multiplying the nitrogen content by 6.25, lipid content was

determined by ether extraction, the ash was determined by combusting the samples in a

muffle furnace at 600°C for 5 hours.

3.8 STATISTICAL ANALYSIS

The data obtained from the feeding trials were analysed using the computer software

Genstat Version 8.1 (2005). Completely randomized design in a one-way ANOVA was

used to calculate the mean. The differences in mean were compared using Duncan’s

multiple range test.

23

CHAPTER FOUR

4.0 RESULTS

The proximate composition of experimental diet (Table 5) shows that moisture was

highest in diet II and lowest in diet III. The crude protein was highest in diet I (47.75%)

and lowest in diet V (35.00%). The crude fat (ether extract) level in the diets were

irregular with diet IV having the highest at 28.12% and VI been the lowest at 25.32%.

The crude fibre level were relatively similar in diets I II and III with diet II and III

having the highest value at 5.43% and also diet IV, V and VI having relatively similar

result with diet IV and V showing the lowest at 2.13%. Ash content was uniform with

8.76% in diet VI making it the highest and 8.22% in diet II making it the lowest. N.F.E

was highest in diet V which was 16.41 and lowest in diet II which was 1.91.

Proximate composition of test fish (Table 6) shows that moisture content was highest in

fish fed with diet I at 8.21% and lowest in fish fed with diets III at 4.73%. Crude protein

level was irregular been highest in fish fed the test diets with 64.20% in diet II and

lowest at 50.17% in diet VI. The fat content were irregular with diet VI been the highest

at 32.11% and diet II being the lowest at 22.75%. Ash content of test fish was highest in

fish fed with diet I at 7.22% and lowest in diet II at 5.42%. Carcass fed with diet II had

the highest crude protein value (64.75% CP) and carcass fed with diet VI had the lowest

value (50.17% CP). When compared with the initial value (52.75% CP), treatments II,

III, V and I increased with 64.75% CP, 63.25% CP, 60.75% CP and 59.50% CP

respectively, while treatment IV and VI with CP values of 50.75% and 50.17%

reduced. Lipid value also varied with treatments been highest in treatment VI and

lowest in treatment II.

24

Table 5: Proximate Composition (%) Of Experimental Diets

TREATMENT

Proximate

composition

I II III IV V VI

Moisture content

(%)

10.44 14.72 10.24 10.75 12.73 13.23

Crude protein (%) 47.75 42.5 45.5 35.5 35.00 36.75

Ether extract (%) 26.12 27.22 26.52 28.12 25.42 25.32

Crude fibre (%) 5.30 5.43 5.43 2.13 2.13 2.22

Ash % 8.52 8.22 8.75 8.32 8.31 8.76

N. F. E (%) 2.07 1.91 3.56 15.18 16.41 13.72

(Source: As analyzed by me)

Table 6: Carcass composition (%) of Heteroclarias post fingerlings fed varying

levels of L. leucocephala seed meal based diets for 56 days.

Initial fish

carcass

TSF

I

TSF

II

TSF

III

TSF

IV

TSF

V

TSF

VI

Moisture

content (%)

10.23 8.21 6.23 4.73 7.25 6.34 5.75

Ash (%) 7.52 7.22 5.42 6.27 6.36 6.72 7.21

Fat (%) 28.21 24.33 22.75 25.11 30.22 25.45 32.11

Crude protein

(%)

52.75 59.50 64.75 63.25 50.75 60.75 50.17

N.F.E (%) 1.29 0.74 0.85 0.64 5.42 0.74 4.76

25

TSF = Test fish carcass composition (Source: As analyzed by me)

The growth response and nutrient utilization data evaluated (Table 7) displayed an

irregular trend with various substitution levels. At all levels of substitution, the weight

gain was different in all the treatments. The highest weight gain was recorded in

treatment I (7.714) that was fed with diet containing 0% leucaena seed meal. This

treatment was not significantly different (P > 0.05) from treatment II but significantly

different from treatment III to treatment IV, treatment V and VI were not significantly

different from each other. There was significant difference in weight gain with

increasing inclusion levels with treatment IV (1.798) having the lowest weight gain.

Table 7: Growth response and nutrient utilization of Heteroclarias post fingerling

fed Leucaena leucocephala seed meal based diets.

TREATMENT

PARAMETER

I II III IV V VI SEM

0% 10% 20% 30% 40% 50%

Mean weight

gain (g)

7.714a 7.312a 5.178ab 1.798c 4.111bc 3.751bc 1.492

Feed intake (g) 22.02a 21.98a 18.06b 16.44b 18.12b 16.54b 1.225

Percentage

weight gain (%)

10.531a 10.08

4a

9.893a 8.610a 7.696a 6.720a 2.067

Specific growth

rate (%)

1.413a 1.500a 1.128ab 0.271b 1.067ab 1.054ab 0.397

Feed conversion

ratio

3.305d 3.742cd 4.517bc 5.735a 4.754abc 5.161ab 0.510

Protein

efficiency ratio

(%)

0.8208a 0.8242a 0.6733a 0.2908a 2.2717a 0.5504a 1.021

Survival rate 93.33 100 100 73.33 93.33 100 -

26

(%)

Figures in each row with the same superscript are not significantly different (P > 0.05)

SEM = standard error of mean (Source: As analyzed by me)

Feed intake in treatment I and II was not significantly different (P > 0.05) from each

other but both were significantly different from treatment III, IV, V, VI meaning that

feed was consumed at different levels of intensification within each treatment.

Treatment I had the highest feed intake of 22.02g and the lowest was recorded in

treatment IV (16.44g).this result indicates that the best intake of feed containing

leucaena was at 10% inclusion level and that with increasing levels from 20% - 50%,

there were similar feed intake as there was no significant difference between treatment

III, IV, V and VI.

Percentage weight gain showed no significant difference (P > 0.05) between all

treatments as this was similar with treatment I (10.531%) being the highest and

treatment VI (6.720%) being the lowest.

Specific growth rate in treatment I and II showed no significant difference (P > 0.05)

from each other but different from treatment III, V, VI and IV. Treatment III, V and VI

were not significantly (P > 0.05) from each other. Treatment II (1.500%) had the

highest specific growth rate with treatment IV (0.271%) having the lowest value.

There was significant difference (P > 0.05) in the feed conversion ratio of all the

treatments indicating that food was converted to flesh at different rate. FCR was highest

in treatment I (3.305) and lowest in treatment IV (5.735). The FCR was uneven with

increasing inclusion levels.

Protein efficiency ratio showed no significant difference (P > 0.05) between all

treatments as this was similar with treatment II (0.8242%) being highest and treatment

IV (0.2908%) being the lowest.

Survival rate was jointly highest in treatment II, III and VI (100%), followed by

treatment V (93.33%) while treatment IV (73.33%) had the lowest survival rate. The

values gotten indicate that inclusion levels did not affect the survival of the fish.

27

CHAPTER FIVE

5.0 DISCUSSION

The growth rate varied with different inclusion level of luecaena seed meal, it was

highest in treatment I (10.531%) and decreased variably as the inclusion level of

leucaena seed meal (LSM) increased (treatment II had 10.084%, treatment III had

9.893%, treatment IV had 8.610%, treatment V had 7.696% and treatment VI had

6.720%). This variation in growth rate that was highest in leucaena replaced diets may

have been as a result of the presence of heat resistance anti-nutrients (Alegbeleye et al.,

2001) which might not have been totally removed with toasting and the non-inclusion

of essential amino acid ‘methionine’ which is known to be limiting in both test

ingredients (soybean and leucaena) (FAO, 1983). This observed pattern could also

probably be a result of persistent consumption of leucaena meals which could retard

animal growth rate as reported by Jones (1997), and further buttressed by Tangendijaja

et al., (1990) who recorded progressive depressed growth rate in rabbit fed increasing

graded levels of leucaena leaf meal based-diet. Nutrient utilization (feed conversion) of

fish decreased as level of LSM inclusion increases in the diets.

The poor nutrient utilization may not be hinged on the fibre content of the test diets as

this was generally less than 10% as recommended by Aladetohun (2008), who stated

that the main factor in the digestibility of feed is the fibre content. In this observation,

this may be as a result of the presence of toxins or nutrient imbalance that is associated

with plant protein source (De Silva and Gunasekara, 1989). The variation in growth and

nutrient utilization by fish with increasing inclusion levels is in agreement with Sotolu

and Faturoti (2009), who reported that the growth and nutrient utilization of diets by

fish decreased as level of LSM inclusion increases in the diets.

Heat treatment is known to detoxify anti-nutrients but retarded growth and nutrients

utilization recorded in this study was in line with the findings of Sotolu and Faturoti

28

(2008), who reported that heat treated leucaena seeds gave lower performance than

leucaena soaked in water and sundried. Heating of the seeds could have also resulted in

the destruction of the amino acid bonds thereby reducing the protein quality of the feed

ingredients. This was supported by Osuigwe et al., (2005) that heating destroys and

reduces nitrogenous compounds in legume seeds. From observation, feed intake was

satisfactory in the first week of the experiment but decreased variably afterwards with

increasing inclusion levels of leucaena seed meal. This is similar to the findings of

Ahmed and Abdelati, (2009) and Atawodi et al., (2008) who reported that Leucaena

leucocephala supplementation progressively decreased appetite in laying hens.

Protein efficiency ratio (PER) was highest in fish fed with 10% LSM meal but it was

similar statistically to values of 0%, 20%, 30%, 40% and 50% LSM inclusion. This is in

conformity with what was stated by Sotolu and Faturoti (2009), who reported that

similarity in the PER of Clarias gariepinus has a direct link with feed intake. All diets

produced different values of fish carcass protein and lipid than initial values with

marginal difference among them indicating different utilization levels of the diets.

These relatively high value of protein can be crude protein could be viewed from the

alongside the work of Alegbeleye et al.,(2001) who reported that effective utilization of

bambara groundnut at varying rates was responsible for variations in Heteroclarias

carcass protein and lipid.

The non-detection of crude fiber in the fish carcass composition was the same in all

treatments and this had been said to be associated with effective utilization of diets

according to Sotolu and Faturoti (2008). Base on this study, it has been observed that

fish with lower LSM inclusion levels had better health status than those of higher LSM

inclusions based on earlier submissions of Svobodova et al., (1991), Alegbeleye

(2005), Ochang et al., (2007) and Sotolu and Faturoti (2009).

29

CHAPTER SIX

6.0 CONCLUSIONS AND RECOMMENDATIONS

In conclusion, the result obtained from this study showed that Diet I with 0% inclusion

of LSM was the best but this was not significantly different from Diet II with 10%

inclusion level which performed best among the other Diet that had LSM present in it.

The low performance of the diets with 10% - 50% inclusion levels of LSM may have

been as a result of partial detoxification of the anti-nutrient (mimosine) present in the

leucaena seed. From the study carried out, the recommended levels of LSM are 0% and

10% for catfish Hybrid (heteroclarias) since they performed better than the other

inclusion levels but since weight gain of fish is what would translate into income for the

fish farmer at the end of the production cycle, 10% inclusion rate of LSM in catfish diet

would produce better and profitable result at present. Cost of fish production is

expected to further reduce if more soya bean meal could be replaced by leucaena seed

meal.

I therefore recommend that:

The commercial planting of Leucaena trees should be encouraged.

Other methods of detoxification such as soaking, boiling and then sun-drying

should be used for removing the anti-nutrient (mimosine) present in it.

Further trials should be carried using other inclusion levels of leucaena seeds.

30

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