THE USE OF Terminalia Superba TO INDUCE OVULATION IN GRAVID FEMALE

Clarias gariepinus

BY

ODUYEBO, BUSAYO OYINLOLA

MATRIC NO 06/0807

SUBMITTED TO

THE DEPARTMENT OF AQUACULTURE AND FISHERIES MANAGEMENT, COLLEGE OF ENVIRONMENTAL RESOURCES MANAGEMENT,

UNIVERSITYOF AGRICULTURE, ABEOKUTA.

A PROJECT REPORT SUBMITTED IN PARTIAL FULFILMENT OF THE AWARD

OF THE DEGREE OF BACHELOR OF AQUACULTURE AND FISHERIES

MANAGEMENT

JUNE, 2011

CERTIFICATION

This is to certify that this project work was carried out by Miss Oduyebo Busayo Oyinlola with

matriculation number 06/0807, in partial fulfillment of the requirement for the award of Bachelor of

Aquaculture and Fisheries Management (B.AQ& FM) of the University of Agriculture, Abeokuta, Ogun

- State and duly supervised by DR.(MRS) N.B IKENWEIWE.

______________________ ____________________

Supervisor Date

DR. (MRS) IKENWEIWE

_____________________ _________________

Head of Department of Aquaculture Date And Fisheries Management

Prof. Y. Akegbejo Samsons

DEDICATION

To the Almighty God, the beginning and the end of everything, He has been my strength, my wisdom

and all I can achieve in life is in Him.

ACKNOWLEDGEMENT

A success story is never achieved by a single person, as no man can do all alone. I appreciate

God Almighty for preserving my life till the end of my stay as undergraduate on the land of UNAAB,

who started the journey and ended it well with me. Unto Him I return all the glory, honor and adoration.

I will not but acknowledge everyone that has contributed in one way or the other to my success

in UNAAB.

My appreciation first goes to my supervisor, a Mother indeed, Dr.( Mrs.) N.B Ikenweiwe for her

full support and advice. To my lovely and caring parents Mr. and Mrs. Oduyebo who took upon

themselves the burden of my success and for all their moral and financial support. I pray that they will

eat the fruit of their labour and that the Lord God will preserve their lives to see many more of their

childrens success in life.

I will be an ingrate if I neglect the impact of this glorious and great people of God upon my life, I

say you are great The Apostolic Church Student Fellowship of Nigeria UNAAB. I will never forget

you all.

Also I appreciate the love of my dearly beloved brothers and sisters, Mr. and Mrs. Orilua,

Sunday, Kolawole, Olumide and Kemi oduyebo. May the Lord bless each and every one of you in Jesus

Name.

Words cannot tell my appreciation to this great friends, who stood by me as a pillar Abosede

Kasunmu,Ogunlade Oluwasegun, Oke Istrael, Amusan Timothy, Adenuga Abiola, Olorunloto Tope and

Yetunde Okelana.I pray that my God will reward you for sincere love towards my life.

I cannot but appreciate all my lecturers in the Department, I love you all.

TABLE OF CONTENT

pages

TITLE PAGE i

CERTIFICATION ii

DEDICATION iii

ACKNOWLEDGEMENT iv

TABLE OF CONTENT v

LIST OF TABLES ix

LIST OF PLATES x

ABSTRACT xi

CHAPTER ONE

1.1 Introduction 1 1.2 Objectives of study 4 CHAPTER TWO

2.0 Literature review 5 2.1 Aquaculture 5 2.2 Cat fish 7

2.2.1 Uniqueness of of Catfish farming 8

2.3 Fish breeding 9

2.3.1 Natural spawning in open water 11

2.3.2 Natural breeding in ponds 12

2.3.3 Spawning in tanks 12

2.3.4 Induced natural spawning in ponds and tanks 13

2.3.5 Artificial fertilization 13

2.4 Seed quality 14

2.5 Medicinal plants 14

2.6 Effect of some plants on animal reproduction 15

2.6.1 Effect of ethanolic extract of Garcinal kola on the fertility of animal 15

2.6.2 Effect of rhizome Curculigo orchioides on the spermatogenetic

activities of animals 16

2.6.3 Effect of Moringa oleifera on the reproduction activities of animals 17 2.7 Terminalia superba 18

2.7.1 Ecology and distribution 19

2.7.2 Uses of Terminalia superba 20

CHAPTER THREE

3.0 Materials and Methods 21

3.1 Experimental site 21

3.2 Experimental diet 21

3.2.1 Feed formulation 21

3.3 Phytochemical screening of Terminalia superba (leaf, back and root) 22 3.3.1 Procedures for Phytochemical screening 23

3.4 Experimental fish 24

3.4.2 Removal of pituitary gland 24

3.4.3 Injection of female broodstock 24 3.4.4 Extraction of milt and fertilization 25

3.4.5 Incubation of eggs 25

3.5 Water quality parameters 25

3.6 Statistical analysis 26

CHAPTER FOUR

4.0 Result 27

4.1 Latency period 27

4.2 physicochemical parameters of water 27

4.3 Phytochemical screening of Terminalia superba 28

4.4 Fecundity 29

CHAPTER FIVE

5.0 Discussion and conclusion 30

5.1 Discussion 30

5.2 Conclusion 31

5.3 Recommendation 31

References 32

Appendix 37

LISTOF TABLES

TABLE PAGES

Table 1 Feed Formulation 22

Table 2 Latency period 27

Table 3 physicochemical of water 27

Table 4 Result of Phytochemical Screening of Terminalia superba 28

Table 5 Fecundity 29

LIST OF PLATES

Page

PLATE 1 Terminalia superba root in grinded form 32

PLATE 2 Stripping of fish 32

PLATE 3 Addition of milt 32

PLATE 4 kaka ban 32

ABSTRACT

The effect of Terminalia superba was tested on the ovulation of a gravid female Clarias

griepinus using two treatments with three replicates each, T1 contains 20% of Terminalia super

root while T2 contains 0% of Terminalia superba which serves as the control experiment.

The result of the experiment reveals a very positive effect of Terminalia superba root on the

Latency period of a gravid Clarias gariepinus which was 4hours 30minutes.

The Data on fecundity of the control (T2) was compared with the treated female C. gariepinus

(T1). The result showed no significant difference, P < 0.005. T2 which serves as the control was

revealed to have the highest fecundity count ( 2583) while the treated fish T1 had the least

fecundity count (2016). This was traced to the high concentration of saponin which is known to

have a negative effect on animal reproduction.

The Phytochemical screening of the plant also established the presence of medicinally active

constituents like Flavonoids(2.64), alkaloid(2.12), phenols(6.35), tannins(3.25), steroid(2.33),

saponin(10.34), glycoside(1.12), phlobatannin(2.64), and anthraquinones(3.56) which suspects it

effectiveness on the fish.

CHAPTER ONE

1.0 INTRODUCTION

Fish and fisheries products are integral part of most societies and make important contributions

to economic and social health and well being in many countries and areas. It has been estimated

that approximately 12.5million people are employed in fishery related activities and in recent

years, global production from capture fisheries has tended to vary between approximately 85 and

90 million tons (Cooke and Cowx, 2004). The products from fisheries are used in a wide variety

of ways ranging from subsistence used to international trade as highly sought after and valuable

items. Despite this enormous importance and value, or correctly because of these attributes, the

worlds fish resources are suffering the combined effects of heavy exploitation and in some

cases, environmental degradation (Bondad-Reantaso, 2000) . The FAO (2000), estimates that in

1999, 47% of the 441 stocks for which some information on status was available were fully

exploited, 18% over exploited,9% depleted and 1% recovery. This pattern is broadly consistent

with similar statistic from other region. For example, the National Marine Fisheries Service of

the United States of America estimated in 1998 that 30% of the stocks in the waters of that

country for which information was available were overfished. In the waters of the European

community, it was estimated that in 1990, 57% of the stock were heavily exploited. Fish stocks

throughout the rest of the globe are likely to be a similar condition to those in these region. There

are many reasons for this unacceptable state of affairs in Fisheries, but the primary reason all

comes down to the failure in fisheries governance in most countries. This is responsible for

declining stock and falling economic returns and employment opportunities. However all too

often, the fisheries manager remains either unaware of the state of the resources, or fails to act

sufficiently as the fisheries slip further and further into decay and crisis or both. Since there is a

gradual depletion of fisheries resources due to over exploitation and mismanagement of our

natural water bodies, there is a need to develop people in the knowledge of aquaculture to meet

the protein requirement of man.

In 2006 the global production from fishing and aquaculture combined, reached approximately

144 million tones, of which 110 million were for human consumption. In 2008, the global

production of fish and fishery products from either capture fisheries or aquaculture, increased by

1percent only from the level in 2007. Aquaculture also increased largely to meet growing

demand; however, the growth may be dampened by high fuel and feed costs which are forcing

many producers to reduce their production. This sector alone accounts for about a third of the

worlds supply of fish products compared to only 4% in 1970 (Atanda, 2007).

Fish is the cheapest source of animal protein and it is available in different forms, it could be

frozen, dried, smoked or fresh. Most fish are "cold-blooded", or ectothermic, allowing their body

temperatures to vary as ambient temperatures change. Fish are abundant in most bodies of water.

They can be found in nearly all aquatic environments, from high mountain streams (e.g., char

and gudgeon) to the abyssal and even hadal depths of the deepest oceans (e.g., gulpers and

anglerfish). At 31,900 species, fish exhibit greater species diversity than any other class of

vertebrates. Among the culturable fish in Nigeria includes C. gariepinus, which is a major

tropical aquaculture species in Africa (Ayinla and Akande 1988) and most popular with fish

farmer and consumers. C. gariepinus commands a very good commercial value in Nigerian

markets (Ayinla et al, 1994). It has been noted that farming is hardly imaginable without the

availability of fish seed ( Chondar 1980). Based on a 1992 United Nations Development Project

(UNDP) assisted base line study, the total annual fingerlings requirement for Nigeria was

250,000 million while the domestic production stood at 7.2 million (Nwokoye et al. 2007).

In fish reproduction under controlled conditions, attempts are made to obtain sperm of the

highest quality and hence to produce the highest possible numbers of good quality seeds. Several

factors that affect fish seeds quality includes different strains, genetics, nutrition, content of feed

and activities of modern agriculture which have introduced several substances such as organic

matter, chemical fertilizer and insecticides into the water used for cultured medium. (Conyurt

and Akhan, 2008).Common practices in hatcheries such as transportation, handling, cleaning,

crowding, use of chemicals, and problems with water quality are stressors that may negatively

influence reproduction (Billard et al 1995).These factors affect fertilization success in artificial

reproduction commonly used for aquaculture species. As a result, low quality fish seeds are

produced. The need for high quality fish seed has necessitated research into various ways of

enhancing fertility to meet the growing demand. However the continuing expansion of

aquaculture requires shifting from synthetic drugs to natural plant. Medicinal plants that were

once considered of no value are now being investigated, evaluated and developed into drugs with

little or no side effects (Adedeji et al.2006). The use of medicinal plants as fertility enhancer in

aquaculture has now being receiving some attention. Dada and Ajiore (2009) used extract of G.

kola seed to enhance fertility in C. gariepinus. Kigelia africana (Lam) Benth, belongs to the

family bignoniaceae. It is abundant in the tropics and is widely used in southern Nigeria as a

herbal remedy for various ailments such as diarrhea, malaria, rheumatism, retained placenta and

dizziness (Gill 1992). Sexual complaints such as infertility, poor libido, sexual asthenia and

impotence are treated with medicines containing the fruits, roots or leaves of K. africana

(Owolabi and Omogbai 2007). K. africana fruit extracts had been used successfully as fertility

enhancing agent in rats (Abioye et al. 2003). It is therefore not out of place to expect a similar

effect on fish.

1.1 OBJECTIVE OF THE STUDY

To check the effect of the root of Terminalia superba base diet on the fecundity of a gravid

female Clarias gariepinus.

To check the effect of Terminalia superba base diet on the latency period of a gravid female

Clarias gariepinus.

CHAPTER TWO

2.0 LITERATURE REVIEW

2.1 AQUACULTURE

There is an ongoing debate about the extent to which aquaculture should be seen as a branch of

Fisheries or as another form of farming. Although basically a non-question, most people consider

aquaculture as a form of animal husbandry, it must be acknowledged that aquaculture is often

viewed in isolation from other farming practices. Food and Agriculture Organization of the

United Nations (FAO 1998) defines aquaculture as The farming of aquatic organisms including

fish, mollusks, crustaceans, and aquatic plants.

Farming implies some form of intervention in the rearing process to enhance production, such as

regular stocking, feeding, protection from predators etc. farming also implies individual or

corporate ownership of the stock being cultivated.

According to Huet (1972) and Bardach et al. (1972) aquaculture is a form of animal husbandry

comparable to poultry or livestock keeping. Fishes are confined in specially, but simply designed

independent enclosures (ponds or cages or tanks) over which the farmer has total control and in

which proper and regular feeding is done. Usually a known number and size of fish seeds are

introduced into the enclosure and reared for short periods, depending on the types of fish, after

which adult fishes are harvested. Fish farming is practiced in ponds, within lakes and reservoirs,

in cages positioned along the course of running water and concrete-block tanks. In all these

cases, excepting in cages, water is impounded and retained against seepage within the enclosure

made from earth of clayey texture or concrete. Raising fish in ponds is about the oldest and most

common form of fish culture practice. Their establishment, mode of construction, size and

classification depend on:

(a) The location, size, topography, geochemistry and accessibility of site,

(b) Quality and quantity of water available,

(c) Type and ultimate size of the fish to be culture,

(d) The ultimate use of the pond

(e) The investment capital available and degree of acceptable risk, and

(f) The scale of the operation

Ponds can be used for the production of fingerlings, table-size fish, bait fish or ornamental fish.

They can also be designed for recreational fishing. Ponds can be built on land unsuitable for

agriculture and integrated with other agricultural enterprises such as poultry, piggery, rabbitary,

snailry and livestock. This is referred to as integrated fish farming and it has numerous

advantages. For example, droppings from poultry and livestock serve as supplementary feed for

fish and organic fertilizer for the pond bottom, as fertilization provides the best means of

increasing fish production in ponds. Aquaculture practices are traditionally ranged on a

continuum from extensive or subsistence aquaculture, to semi-intensive aquaculture and

intensive aquaculture. In extensive aquaculture, cultured organisms are sometimes collected

from the wild, kept at low densities and are not actually fed, but the culture media may be

fertilized in order to enhance the production of natural food (natural production). In semi-

intensive aquaculture, cultured organisms are kept at higher densities than in extensive culture.

Agricultural by-products are regularly fed as supplementary feed, and the culture media are

usually fertilized in order to enhance natural production. In intensive aquaculture, culture

organisms are nearly always reproduced in specially-designed hatcheries, kept at high densities

and fed several times per day. The feeds are palletized and nutritionally complete, so that fish

production is independent of natural production.

2.2 CAT FISH

Clarias is a genus of catfishes (order Siluriformes) of the family Clariidae, the air breathing

catfishes. The name is derived from the Greek chlaros, which means lively, in reference to the

ability of the fish to live for a long time out of water (Froese et al 2007) Clarias has been found

to be paraphyletic. It has been found that a species of Heterobranchus (H. longifilis) clusters

deeply inside the Clarias group (Nkwengulila, et al 2008). They are found in inland waters

throughout much of the Old World, and are one of the most widespread catfish genera in the

world Heok Hee (2001). The genus is found in Southeast Asia and East Asia westwards through

India and the Asia Minor to Africa (Kelvin,et al 1999). The diversity of these catfishes is highest

in Africa. Some (notably the Walking catfish) have become pest species where they have been

accidentally introduced. Clarias catfish and primarily Clarias batrachus (walking catfish) has

been introduced to many different areas of the world where they are causing problems for the

native wildlife. The effect of introduction of Clarias catfish varies from area to area but as they

are predatory fish they often impact the local wildlife by eating other fish, birds and amphibians.

In Florida the fish is causing problems by invading aquaculture farms and prey on the fish

cultivated there. Countries where one or several Clarias species have been introduced include

Indonesia, USA, Hong Kong, Taiwan, China, UK, Papua New Guinea, Guam, Taiwan, Thailand

and Cuba.

2.2.1 UNIQUENESS OF CATFISH FARMING

Catfish farming occupies a rather unique status in global aquaculture. The uniqueness of this

farming system could be summarized as follows:

It is a farming system that is capable of producing, on average, 300 - 400 tones /ha/crop; one of

the highest recorded for any primary production sector in the world.

The industry essentially occupies approximately 5,400 ha of land but produces, for example, as

much as 65 % of the total aquaculture production in Europe. It includes a range of farming

scales, from small, household scale through to large, industrial scale enterprise.

It provides many livelihood opportunities to poor rural communities, particularly women (in the

processing sector in particular), significantly bypassing that seen elsewhere in the aquaculture

industry around the world.

It is a farming system that is mostly conducted in earthen ponds of 4 - 4.5 m depth, with regular

water exchange from the Mekong River and/or its tributaries.

The farming system is blessed with an adequate water supply through the year, but is also

obligated to ensure that the same water source is not overly nutrient loaded through effluent

discharge bringing about negative impacts on all downstream users of this common, valuable

resource.

It is a farming system that for all intents and purposes is both horizontally and vertically

integrated, with specialized hatchery production, fry to fingerling/ nursery rearing and grow-out

phases; even including various post-harvest activities including processing for some larger,

industrial-scale producers.

It is a farming system from which the produce is almost totally destined for export, being an

acceptable and affordable substitute for white fish, particularly for the western palate/ taste.

2.3 FISH BREEDING

Fish have evolved to reproduce under environmental conditions that are favorable to the survival

of the young. Long before spawning, seasonal cues begin the process of maturation. In many

fish, this can take up to a year. When the gametes have matured, an environmental stimulus may

signal the arrival of optimal conditions for the fry, triggering ovulation and spawning. Examples

of environmental stimuli are changes in photoperiod, temperature, rainfall, and food availability.

A variety of sensory receptors detect these cues, including the eye, pineal gland (an organ in the

dorsal part of the forebrain that is sensitive to light), olfactory organs, taste buds, and thermo

receptors. The hypothalamus, located at the base of the brain, is sensitive to signals from sensory

receptors and releases hormones in response to environmental cues. Principal among these

hormones are gonadotropin releasing hormones (GnRH), which travel from the hypothalamus to

the pituitary gland. The pituitary is responsible for a wide variety of functions, including growth

and reproduction. Certain cells of the pituitary receive GnRH and release gonadotropic hormones

into the bloodstream. The gonadotropic hormones travel to the gonads, which synthesize steroids

responsible for final maturation of the gametes. Maturation of the egg is a long process that

involves complex physiological and biochemical changes. One important step, vitellogenesis, is

a process in which yolk proteins are produced in the liver, transported to the ovary, and stored in

the egg, resulting in tremendous egg enlargement. The yolk is important as a source of nutrition

for the developing embryo. Also critical are germinal vesicle migration and germinal vesicle

breakdown (GVBD). Before it migrates, the germinal vesicle, or nucleus, is located at the center

of the egg in an arrested stage of development. At this stage, the egg is physiologically and

genetically incapable of being fertilized, even though it has the outward appearance of a fully

mature egg. When conditions are appropriate for final maturation, nuclear development resumes,

and the germinal vesicle migrates to one side. Finally, the walls of the germinal vesicle break

down, releasing the chromosomes into the cell. The maturity of eggs can be determined using

biopsy techniques. Eggs are removed from the ovaries, cleared with a prepared solution, and

viewed under a microscope. In mature eggs, the migration of the germinal vesicle to the side of

the cell will be complete. After the egg has matured, a class of compounds called prostaglandins

is synthesized. These stimulate ovulation, which is the rupture of the follicle cells that hold the

egg. The egg is then released into the body cavity or ovarian lumen, where it may subsequently

be released to the outside environment. Following ovulation, the viability of the eggs can

decrease rapidly. Fish with gametes that have not yet been released by the gonads are called

green. The term ripe is used to describe fish with gametes that have been released from the

ovary into the ovarian lumen. Ripe fish can be stripped, green fish cannot.

According to Atanda (2007). The various ways and methods by which fish seed are produced in

Nigeria include

the following:

1) Natural spawning in open waters and rivers

2) Natural breeding in ponds

3) Spawning in tanks

4) Induced natural spawning in ponds and tanks

5) Artificial fertilization hatchery production and management

6) Genetic manipulation.

2.3.1 Natural spawning in open water

This describes the means by which fish breed in the wild to produce their seed. Different fish

species usually choose different places in the aquatic environment for breeding. The reasons for

the choice are not always fully known and depend on behavior and adaptation of the fish species.

For example, some breed on stones at the bottom of the water. Others make holes in the benthos

for breeding; others lay their eggs in holes, on grass, or in a foam nest on the surface of the

water. Some scatter their eggs in the water while others collect and brood the fertilized eggs in

their mouths. In nature, the cues or signals that induce spawning are numerous, e.g. flooding

(increase in water volume), rain events, changes water temperature, increase in food, etc. As the

natural aquatic environment is full of hazards or perils for the young fish, fish have evolved

varying degrees of parental care to help reduce the level of mortality for the young. Some care

for the eggs only, moving away some after the eggs are hatched. For others, protection continues

until the young are able to fend for themselves. Cichlids, i.e. some tilapias, have elaborate

parental care, including carrying the young in the mouth at any approach of danger. As a rule,

species with elaborate parental care lay relatively few eggs, while clupeids and catfishes (e.g.

Chrysichths nigrodigitatus, Clarias gariepinus, Heterobranchus bidorsalis) with little parental

care produce by far more numerous eggs, as a way of ensuring that some, at least, will survive to

adulthood. Fry, fingerlings and juveniles are collected from open waters by artisanal fishermen

using baskets, fine mesh nets and cane traps. Collection is always easier during schooling

(especially for Heterotis niloticus). Fish farmers purchase the collected seed and transfer them

directly into ponds. Many fish farmers in the northern and north central zones of the country

depend largely on wild fingerlings from the vast inland water resources such as lakes and

reservoirs. However, wild stocking of fish farms has almost disappeared in the south (Atanda,

2007).

2.3.2 Natural breeding in ponds

Some fish farms have specially dedicated ponds for breeding. These are usually smallto medium-

sized ponds (e.g. 100-300 m2). Depending on the species of fish, the pond bottom may be

modified to simulate the desired depth preferred by the fish in nature. For example, one end of

the pond may be shallower, or the shallow end may be in the middle potion to facilitate recovery

of the parent after spawning. Substrate (e.g. grass, mat) for egg attachment may also be provided.

During the breeding season, male and female may be placed either at random or in definite ratio

in the breeding pond and left to spawn naturally after which the parents are removed and the fry

nurtured to fingerlings. Fry and fingerling recovery is usually very low and in most cases hardly

greater than 5 percent of hatchlings. This is the most common method for tilapia seed

production.

2.3.3 Spawning in tanks

Fish spawning can also be carried out in outdoor or concrete or plastic tanks. Breeding tanks can

be in the form of small aquaria (glass or plastic) or large fiberglass and concrete tanks. Brood

stocks may be induced or just allowed to freely spawn by mutual stimulation resulting from

proximity of the male and female parents. Outdoor spawning tanks are always covered with net

mesh for protection against predation by insects, birds and reptiles.

Most commercial fish farms, which may not have a complete hatchery complex, always, have

one to four concrete tanks dedicated for breeding/spawning, where fry can be harvested to

earthen nursery ponds and then High-tech water recirculation system transferred to grow-out

ponds. Depending on the type of management, up to ten percent of hatchlings can be harvested

as fry.

2.3.4 Induced natural spawning in ponds and tanks

A further step in the controlled breeding of fish is to induce the brood stock through hormonal

stimulation before pairing them up in breeding tanks or ponds. Natural production is inadequate

due to the low survival of hatchlings caused by factors such as predation, poor water quality,

pests and parasites in the natural environment. Apart from the fact that in nature most cultured

species, especially Clarias gariepinus, do not spawn year-round, a better control of fry

production is required. At the inception of induced breeding trials in Nigeria in the 1980s, the

following hormones are used: carp pituitary (fresh and acetone-dried) Desoxy cortico sterone

acetate (DOCA), human chronic gonadosterone (HCG) and fresh catfish pituitary. With more

research and networking, cheaper and more effective synthetic hormones are now in use, the

most popular being Ovaprim used in catfish breeding. Various other derivatives are also

available, especially those from Asia such as Suprefact and Motilium from Thailand.

2.3.5 Artificial fertilization

Artificial propagation through hormonal treatment and stripping under controlled environmental

condition in the hatchery had become a necessity to ensure mass production of fry and

fingerlings of the African catfish. This is the current status of fish seed production in Nigeria.

The procedure has been generally standardized as a synthesis of local experience, trial and error

as well as from literature and largely from the Manual on Catfish Hatchery and Production

(Kamthorn and Miller, 2006) which has become the field guide for Nigeria catfish breeders.

2.4 Seed Quality

The increase of seed demand has created concerns on seed quality. Striped catfish seed quality is

highly influenced by the farmers knowledge on brood stock quality management in hatcheries.

Several studies have shown that brooders are induced to spawn many times each year, the rate

that brooders are added/changed to the brood stock population is relatively low, brooders often

originate from the same source/family (appearing in 90% hatcheries); and cross breeding

between males and females are undiversified. Hien (2008) reported that 21.1% of grow-out

farmers found that the quality of seed stocks seems to have declined over the years.

2.5 MEDICINAL PLANTS

The use of medicinal plant all over the world predict the introduce of antibiotic and other modern

drugs medicinal plants are plants that have chemoprotective and or therapeutic effect on ailment

(Mantle et.al; 2000)

The medicinal properties of plants are mainly attributes to the presence of Flavonoids, but mal

also be influenced, but may also be influenced by their organic and inorganic compounds like

coumarin, phenolic acid antioxidant. Micro nutrient like Cu, Mn, Zn. It is known that plants

accumulate antioxidant chemical e.g Flavonoids as secondary metabolites through evolution as

natural means of surveying in a hostile environment (Monarch et.al; 2004). In recent years, there

has been a gradual revival of interest in the use of medicinal plants in developed and developing

countries because herbal medicines have been reported to be safe and without any adverse side

effect especially when compared to synthetic drugs (Neetu and Meenakshi; 2003)

2.6 EFFECT OF SOME PLANTS ON ANIMAL REPRODUCTION

2.6.1 EFFECT OF ETHANOLIC EXTRACT OF Garcinia kola ON THE FERTILITY OF ANIMALS

In relation with different concentration of Garcinia kola in the feed of the animals e.g Clarias

gariepinus .It causes alteration and degenerative change such as cytoplasm shrinkage, rupturing

of cell membrane and different vacuole sizes. The presence of ethanolic extract in the feed of

Clarias gariepinus shows increase in the egg size (diameter).This shows that the ethanolic

extracts of Garcinia kola seeds possesses promising infertility property which can be exploited

in fish seeds production under hatchery condition. It is therefore recommended that the dietary

concentration of ethanolic extracts of Garcinia kola seeds between 0.25 and 0.59kg feed can be

tolerated by Clarias gariepinus and may be included in the diets for fertility enhancement

because this will minimize the total dependence on synthetic drugs as fertility enhance .(Dada et

al, 2009) The presence of ethanolic extract of Garcinia kola in the diet of a male wistar rats

causes increment in the diameter of their seminiferous tubule.(oluyemi et al, 2007). It also

increases the peripheral testosterone levels in wistar rats (Akpantah et al, 2005). There was

recorded increase in the sperm count of wistar rats treated with ethanolic extract of Garcinia kola

for the period of 8 weeks. (Adesanya et al, 2007).

Other study in man, has shown that Garcinia kola helps man with infertility, with improvement

in male fertility especially sperm characteristics. This might be as a result of biflavonoid and

xanthone in the plant. These compounds are protein anti oxidants which are capable of

increasing the production of oestrogen, the key hormone involve in the production and

maturation of eggs in the ovary.(Adesanya et al, 2007)

2.6.2 EFFECT OF RHIZOME OF Curculigo orchioides ON THE SPERMATOGENETIC ACTIVITIES

OF ANIMALS.

Albino rats of either sexes weighing 120-150g were feed on standard diet and water ad libitum.

The animals were housed at worm temp (242oc) on a reversed day-night cycle (06.00hrs to

18hrs). Rhizomes of Curculigo orchioides was dried and made into powder. It was defatted by

extraction with petroleum ether (60-80c). The defatted plant materials was then extracted with

ethanol (95%), and dried under vacuum (4.08% w/w). The extract was administrated to the

albino rats subcutaneously for several days, like 30 days.

The extract treated rat showed pronounced effects in term of testis weight and histological

alterations. Since the weight and the size of the testis was greater in the extract treated rats the

seminiferous tubule showed greater diameter. The germinal epithulum cells appeared to be

hyperactive. Large numbers of different cells at different stage of spermatogenesis were evident.

Lumen of the seminiferous tubules had enormous number of spermatozoa. Sertoli cells were

enlarged highly processed and rich in nutrients as evidenced by highly granulated cytoplasm.

This was the normal response of the sertoli cells when they were in readiness for providing

nutritional supplement to large number of spermatozoa (Majumlar 1995). The leg dig cells

showed hypertrophy with enlarged nucleus and darkly stained cytoplasm. Increment in the

volume of cells and nucleus was strongly suggestive of steroid synthesis under the direct or

indirect influence of the drug. Almost all the tubules were over crowded with sperm bundles. In

some tubule, spermatids were found scattered amidst spermatozoa. The blood vessels of testis

were slightly dilated. Increase spermatogenesis was evident by high number of spermatozoa in

the seminiferous tubules and which is evident by increase spermatogenic elements. This also

shows that there is increase in attraction of male towards the female (Chauhan N.S and Dixit

U.K, 2008).

2.6.3 EFFECT OF Moringa oleifera Lam ON THE REPRODUCTION ACTIVITIES OF

ANIMALS.

Male mice of 8 weeks old weighing 28-32g after proper quarantine services were gauged with

0,0.5,5and 50 mg/30g body weight /day of Moringa oleifera lam leaves hexane fraction for 21

days with tap water used. The administration of Moringa oleifera lam hexane fraction at any

dose did not alter the body weight of the animals. The weight of epididymides in all the Moringa

treated mice was significantly increases in contract to the control mice. Moringa oleifera lam

extract at the dose of 50mg/30g also significantly induced the weight of the seminal vesicle of

the male mice. The weight gained may signifies the effect of Moringa oleifera as reported in

Bretinda. (Sudwa et al, 2007) and Lepedium meyenii (Gonzales et al, 2001) and reflect activation

of spermatogenesis as a result of the presence of elongated spermatids in the somniferous

tubules. The study shows that the body weight of the Moringa oleifera treated animals remained

unchanged which shows that the doses selected did not extent any harmful effect and the

metabolic processes of the treated animals were normal. The Moringa oleifera lam in enhancing

male reproductive is clearly manifested in all the treated mice. However, mice administered with

the high and medium doses of the plant extract are reproductively superior to those that were

given low doses. The testicular and epididymal weights, relative maturity ratings, lumen

formation and somniferous tubules diameter shows that hexane fraction of Moringa oleifera has

reproductive effect on the male mice (Libibeth and Gloria,2010).

It has also been discovered in the work by the bureau of plant industry (BPI) that a steady diet of

Moringa fruits boots the sperm count of men thus improves their chances of fertilising an egg

(Cabacumgan, 2008). In India the Moringa fruit is said to increase sexual libido in male.(Serrano

and Pocsidio, 2008) reported an increase in the sperm count in male mice when 1%

concentration of Moringa ethanol leaf extract was administered subcutaneous for two weeks.

2.7 TERMINALIA SUPERBA

Terminalia superba is a large tree in the family Combretaceae, native to tropical Western Africa.

It is also called black korina, limba or white afara (English name), Frake (French name), Akom

(Spanish), Mwalambe (Swahli),Afa or Afara(Yoruba name). Terminalia superba is a large tree,

up to 50 m tall and 5 m in girth, bole cylindrical, long and straight with large, flat buttresses, 6 m

above the soil surface; crown open, generally flattened, consisting of a few whorled branches.

Bark fairly smooth, graying, flaking off in small patches; slash yellow. Root system frequently

fairly shallow, and as the tree ages the taproot disappears. Buttresses, from which descending

roots arise at some distance from the trunk, then support the tree. Leaves simple, alternate, in

tufts at the ends of the branches; deciduous, leaving pronounced scars on twigs when shed.

Petiole 3-7 cm long, flattened above, with a pair of sub opposite glands below the blade; lamina

glabrous, obviate, 6-12 x 2.5-7 cm, with a short acuminate apex. Nerves 6-8 pairs: secondary

reticulation inconspicuous. Inflorescence a 7-18-cm, laxly flowered spike, peduncle densely

pubescent; flowers sessile, small, and greenish-white; calyx tube saucer shaped, with 5 short

triangular lobes. Petals absent. Stamens usually twice the number of calyx lobes (usually 10), in

2 whorls, glabrous; filaments a little longer than calyx; intrastaminal disc annular, flattened, 0.3

mm thick; densely woolly pubescent. Transversely winged, sessile, golden-brown smooth nut,

1.5-2.5 x 4-7 cm (including the wings). Nut without the wing about 1.5 x 2 cm when mature,

usually containing 1 seed. The generic name comes from the Latin terminalis (ending), and

refers to the habit of the leaves being crowded at the ends of the shoots.

2.7.1 Ecology and distribution

Native to West tropical Africa from Sierra Leone to western Congo and northern Angola; planted

in plantations both within and outside its natural range, e.g. in South and Central America,

central and eastern Africa, Hawaii, Fiji and the Solomon Islands. Within the Malesian region,

trials have been carried out in Sabah, Kalimantan and the Philippines.

T. superba is essentially a tree of deciduous forest and sheds its leaves in the dry season. It is

characteristic of tropical high secondary forest areas with a dry season of about 4 months, but it

does not respond well to long dry spells, especially on sandy soils. The species is especially

plentiful at some distance from the coast, but it gains at the expense of the rainforest following

clearances. The tree will withstand occasional flooding. It is frequently struck by lightning,

presumably because of its dominant position in the forest. It is very fire sensitive. However, its

wide spread owes a great deal to the activities of man and to its pioneering characteristics; light

demanding, wide crown and production of regular quantities of viable seed.

Geographicdistribution

Native : Angola, Benin, Cameroon, Central African Republic, Congo, Cote d'Ivoire, Democratic

Republic of Congo, Equatorial Guinea, Gabon, Ghana, Guinea, Liberia, Nigeria, Sierra Leone,

Togo

Exotic : Argentina, Bolivia, Brazil, Burkina Faso, Chile, Colombia, Costa Rica, Ecuador, Fiji,

French Guiana, Guatemala, Guyana, Honduras, Indonesia, Kenya, Malaysia, Mexico, Nicaragua,

Niger, Panama, Paraguay, Peru, Philippines, Solomon Islands, Surinam, Tanzania, Uganda,

United States of America, Uruguay, Venezuela, Zimbabwe

Altitude: 150-1 000 m, Mean annual temperature: 20-28 deg. C, Mean annual rainfall: 1 000-1

800 (3 000) mm Soil type: It grows best on rich, well-drained alluvial soils, but is also found on

other types such as lateritic sands, gravel and clays, lava, black basaltic clays and crystalline

soils. T. superba reaches sexual maturity late and at variable ages, for example 15 years in Cote

dIvoire and 23 years in Congo. The dates of refoliation and flowering are closely correlated;

flowering, which lasts for 2-5 weeks, takes place either as the new leaves are appearing or

immediately afterwards. Rarely, 2 periods of flowering may occur if there are 2 deciduous

periods. Terminalia has an effective system of self-incompatibility. Various insects

(Coleopterans, Dipteral, Hemiptera, Hymenoptera and Lepidoptera) pollinate flowers. Fruit

develops during the rains and mature at the onset of the dry season to coincide with the leafless

period; the duration of fruiting varies from 6 to 9 months. If 2 dry seasons occur, the maximum

seed production occurs in the longer of them. Terminalia trees show interprovenance variability

with regard to early shedding of leaves, early shedding being negatively correlated with vigour.

2.7.2 USES OF TERMINALIA SUPERBA

The wood is either a light ('white limba') or with dark stripes ('black limba' or 'korina') hardwood.

Used for making furniture and musical instruments and prized for its workability and excellent

colour and finish. The most famous example of its use in guitars is when it was used by Gibson

in producing their now highly sought-after Flying V and Explorer guitars in 1958. When finished

in a clear coat, 'White Limba' results in an attractive light golden colour.

Contrary to popular belief, it is not rare and expensive due to overharvesting and there is plenty

of supply due to efforts in the 1950s to preserve natural supply of the wood. This species is

reported to be relatively secure, with little or no threat to its population within its natural growth

range, according to the World Conservation Monitoring Center in 1992.

CHAPTER THREE

3.0 MATERIALS AND METHODS

3.1 EXPERIMENTAL SITE

The experiment was carried out at the Hatchery unit of the Department of Aquaculture and

Fisheries Management, University of Agriculture, Abeokuta.

3.2 EXPERIMENTAL DIETS

Terminalia superba root was collected from Forestry Reserve of Forestry Department,

University of Agriculture, Abeokuta Nigeria, sundried and milled to powder. The test Ingredient

(Terminalia superba) was added to the feed concentrate at 20% levels of inclusion to form the

experimental diets. 0% level served as control. The diet contains 40% Crude Protein as shown in

Table 1.

3.2.1 FEED FORMULATION

The root of the plant was sun dried for two weeks after which it was grinded and mixed together

with other ingredients like maize, soya bean, groundnut cake

Table 1 Percentage of feed ingredients in the Experimental diet

Ingredients Percentage (%)

Fish meal 22.8

Blood meal 19.5

Soybean meal 17.2

Dried Brewer Grain 15.0

Bone meal 1.50

Maize 20

Fish premix 1.0

Methionine 1.0

Lysine 1.0

Oil 1.0

Total 100

3.3 PHYTOCHEMICAL SCREENING OF TERMINALIA SUPERBA (ROOT)

Fresh roots of Terminalia superba collected were taken to the biochemistry laboratory, UNAAB

for analysis. The root were oven dry for three (3) days, powered and tested for the presence of

tannin, alkaloid, Flavonoids, phenol, glycosides, steroids, phlobatannin, saponin, cardenolides

and anthraquinones.

3.3.1 Procedure for Phytochemical Screening

Alkaloids : 5cm3 of 1% HCl was added to 3cm3 of the extract in a test tube. The mixture was

heated for 20minuutes and cooled; 1ml of the filtrate was drops of piric acid solution. Turbidity

or precipitation indicates the presence of alkaloids.

Tannins : 1cm3 of freshly prepared 10% KOH was added to 1cm3 of extract, a dirty while

precipitate indicates the presence of tannins.

Phenolic : 2 drops of 5% FeCl3 was added to 1cm3 of the extract in a test tube, a green

precipitation indicates the presence of phenolics.

Glycosides : 10cm3 of 50% H2SO4 was added to 1cm3 of the extract, the mixture was heated in

boiling water for 15minutes. 10cm3 of Fehlings solution was added and the mixture boiled. A

brick-red precipitate indicates the presence of glycosides.

Flavonoids : 1cm3 of 10% NaOH was added to 3cm of the extract, a yellow coloration indicates

the presence of Flavonoids.

Steroids : 5 drops of concentrated H2SO4 was added to 1cm3of the extract, red coloration

indicates the presence of steroids.

Phlobatannins : 1cm3 of the extract was added to 1% HCl, a red precipitate indicates the

presence of phlobatannins.

Saponins: 0.5g of crude powder was shaken with water in a test tube and it was warmed in a

water bath and the persistent of froth indicates the presence of saponins.

Anthraquinones: 0.5g of crude powder was shaken with 10 ml of benzene and was filtered 0.5

ml of 10 % ammonia solution was added to the filtrate and the mixture was shaken well and the

presence of the violet color in the layer phase indicated the presence of the anthraquinones.

3.4 EXPERIMENTAL FISH

Six sexually matured female broodstocks were collected as a residual of a IFSERAR project by

DR. MRS IKENWEIWE . the fishes had been subjected to two different diets with three

replicate each. The broodstocks were subjected to two experimental diets at 3% body weight

daily in a concrete pond for eight weeks. Treatment one(T1) contains 20% Terminalia superba

root (TR) while T2 which serves as the control experiment contains 0% TR. At the end of the

eight weeks of conditioning, they were subjected to artificial propagation to check the effect of

the effect plant of the plant on the latency period and the fecundity of the fish. The average

weight of the broodstocks was 1.20.8kg.

3.4.1 Removal of pituitary gland

The pituitary gland is a round yellowish organ located under the brain of the fish, the pituitary

gland was collected a day prior the experiment by sacrificing the male fish of the same weight

with the female (1.2kg) broodstock. The pituitary gland was removed through the ventral region

(from the bucal cavity), the mouth of the fish was cut off and the upper roof borne below the eye

was removed which expose the pituitary gland in a capsule form under the brain. After

collection, the pituitary gland was stored in acetone till the next day (the acetone which was

poured at inception was change once after eight hours and replaced with fresh one).

3.4.2 Injection of Female broodstock

The injection was carried out in the morning at 9:45am, the preserved pituitary gland was

pulverized in a proclaim mortar, mixed with of physiological salt solution (9 grams of common

salt was mixed with 1litre of clean water). The dosage used was 1ml of pituitary to 1kg of fish.

Injected fish were kept in separate tanks.

3.4.3 Extraction of milt and Fertilization

At about 2:30pm in the evening, the brood stocks were ready to be stripped; this was detected by

the gushing out of eggs at a slight press of the anal region. Two male catfish each were sacrificed

for T1 and T2, their abdomen was cut opened and the milt was extracted and cleaned with tissue

paper. The matured eggs from the female catfish were stripped into clean, dry bowls, weighed

and the milt was added to the bowl by using a clean blade to pierce the tinny lining of the milt.

The eggs was stripped into separate dry bowls, the milt was mixed gently with the eggs with a

plastic spoon for two minutes. Clean water was added to the mixture to activate the sperm and

fertilize the eggs, decanted and incubated in an improvised incubator.

3.4.4 Incubation of egg.

The eggs were incubated using a fibre glass tank of the dimension 21.20.5m3, laid inside was a

kakaban made with net and taped with PVC pipes at the edge, it also consist an aerator made by

using a hot nail to drill the sides of a PVC pipe. Before starting the experiment the kakaban and

the tank were disinfected with salt and filled with clean and well aerated water to 1/3rd of it

depth. The fertilized eggs were spread on the kakaban to ensure a thin layer of egg. Water was

made to flow through the tank in such a way that the pressure was not much and the inflow of

water was regulated to the outflow of water. After 18-24 hours, the hatchlings were seen

swimming around the edges and sides of the tank.

3.5 WATER QUALITY PARAMETERS

Essential physico-chemical parameters such as the Dissolved Oxygen, Temperature, pH and

Turbidity was monitored in the course of the experiment using a multi-purpose measuring meter.

3.6 STATISTICAL ANALYSIS

Data collected was subjected to analysis of variance (ANOVA) using SPSS statistical package.

The level of significant of means from each treatment was determined using Least Square

Method (LSD).

CHAPTER FOUR

4.0 RESULT

As shown in the table below, the latency period of the female Clarias gariepinus was 4hours

30minutes.

Table 2 LATENCY PEROID

TIME OF INJECTION TIME OF STRIPPING LATENCY PERIOD

9:45 10:45am 2:30 - 3pm 4hours 30 minutes

TABLE 3 PHYSICO CHEMICAL PARAMETERS OF THE WATER

PARAMETER RANGE

Dissolved Oxygen 3.28 3.50

pH 6.9 7.2

Temperature 2 8.1 28.9

Table4 shows the Phytochemical screening of the Terminalia superba leaf, root and bark.

TABLE 4 PHYTOCHEMICAL SCREENING OF Terminalia superba.

Phytochemical

Terminalia

superba(Root)

concentration

Terminalia

superba(Bark)

concentration

Terminalia

superba(leave)

concentration

Alkaloids 2.12 1.36 1.34

Tannin 3.25 2.53 2.61

Phenol 6.35 4.40 4.38

Glycoside 1.12 0.65 0.62

Flavonoids 2.64 1.80 1.86

Steroids 2.33 1.75 1.74

Phlobatannin 1.81 1.15 1.10

Saponin 10.34 9.53 9.31

Anthraquinones 3.56 2.20 2.13

The below table shows the fecundity of the three treatments

Table 5 FECUNDITY

Replicates Treatment 1 Treatment 2 Treatment 3

I 2016 1270 1410

II 1404 1450 1620

III 2583 1920 1457

Total 6003 4640 4487

Mean 2001a 1547a 1496a

CHAPTER FIVE

5.0 DISCUSSION AND CONCLUSION

5.1 DISCUSSION

The study carried out on the plant samples revealed the presence of medicinally active

constituents like Alkaloids, Flavonoids, saponin, tannins, phenol, glycoside, steroid,

phlobatannin, cardenolides and anthraquinones.

The female broodstocks were injected in the morning at 9:45am at an ambient temperature of

30oC and water temperature of 28.2oC, the stripping was done between 2:30-3pm in the evening.

The difference between the time of stripping and time of injection is 4hours 30minutes which

was the latency period. In an ideal situation, the normal latency period of Clarias gariepinus

when using a synthetic hormone is between 8 and 12 hours which also depend on the

temperature. The latency period of a broodstock is said to be faster when a synthetic hormone is

used than when a natural hormone is used. The 4hours 30minutes latency period reveal the

positive effect of Terminalia superba root on the broodstock even when a natural hormone was

used.

The Data on fecundity of the control (T2) was compared with the treated female C. gariepinus

(T1). The result showed no significant difference, P < 0.005. T2 which serves as the control was

revealed to have the highest fecundity count ( 2583) while the treated fish T1 had the least

fecundity count (2016). This could be traced to the high concentration of saponin at the root of

Terminalia superba than other parts like the leaf and bark. Benei 1990 discovered that saponin

have a negative effect on animal reproduction which have been ascribed to it abortifacient, anti-

inplantation and anti- zygotic properties. Saponin-rich extract injected into female rat stimulated

uterine growth, lowered luteinizing hormone performance and growth rate.

5.2 CONCLUSION

The result shows that the root of Terminalia superba induced breeding on gravid female of

Clarias gariepinus.

5.3 RECOMMENDATIONS

I therefore recommend the use of Terminalia superba root to induce breeding on gravid female of

Clarias gariepinus, but care should be taken on the inclusion level because of the concentration

of saponin at the root than every other part of the plant.

PLATE I (Terminalia superba root) PLATE II (Stripping of eggs)

PLATE III(Addition of milt to egg) PLATE IV(Kaka ban)

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APPENDIX Descriptives

fecudity_TR

N Mean Std. Deviation Std. Error

95% Confidence Interval for Mean

Minimum Maximum Lower Bound Upper Bound

treatment 1 3 2.0000E3 500.00000 2.88675E2 757.9311 3242.0689 1500.00 2500.00

treatment 2 3 1.5467E3 335.60890 1.93764E2 712.9679 2380.3654 1270.00 1920.00

tretment 3 3 1.4957E3 110.21040 63.63001 1221.8888 1769.4445 1410.00 1620.00

Total 9 1.6808E3 389.23379 1.29745E2 1381.5862 1979.9694 1270.00 2500.00

ANOVA

fecudity_TR

Sum of Squares df Mean Square F Sig.

Between Groups 462464.222 2 231232.111 1.851 .237

Within Groups 749559.333 6 124926.556

Total 1212023.556 8

Multiple Comparisons

Fecundity TR LSD

(I) group (J) group Mean Difference (I-J) Std. Error Sig.

95% Confidence Interval

Lower Bound Upper Bound

treatment 1 treatment 2 453.33333 2.88590E2 .167 -252.8217 1159.4884

tretment 3 504.33333 2.88590E2 .131 -201.8217 1210.4884

treatment 2 treatment 1 -453.33333 2.88590E2 .167 -1159.4884 252.8217

tretment 3 51.00000 2.88590E2 .866 -655.1551 757.1551

tretment 3 treatment 1 -504.33333 2.88590E2 .131 -1210.4884 201.8217

treatment 2 -51.00000 2.88590E2 .866 -757.1551 655.1551