this work is all about using plant extract instead of synthesized hormone for spawning

1. i
ABSTRACT
The effect of Terminalia superba was tested on the ovulation of a gravid
female catfish (Clarias gariepinus) using four treatments with three
replicates each, D2, D3, and D4 contains 50, 100, 150g/kg of T. superba
root respectively while DI contains 0.00m/kg of T. superba which
serves as the control experiment. The result of the experiment reveals a
very positive effect of T. superba root on the Latency period of a gravid
C. gariepinus which was 4hours 30minutes. The Data on fecundity of
the control D1 was compared with the treated female C. gariepinus D2,
D3, and D4. The result showed no significant difference, P < 0.05. D1
which serves as the control was revealed to have the lest fecundity count
( 1496) while the treated fish D2, D3, and D4 had the highest fecundity
count (2001, 1808 and 2331) respectively. This was traced to the
presence of flavonoid and xanthrone .The Phytochemical screening of
the plant also established the presence of medicinally active constituents
like Flavonoids, alkaloid, phenols, tannins, steroid, saponin,

2. ii
glycoside, phlobatannin, and anthraquinones, which suspects its
effectiveness on the fish.

3. 1
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 (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.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 sensitize people
in the knowledge of aquaculture to meet the protein requirement of man.

4. 2
Aquaculture production is on the increase and the growing fish 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 world’s 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. 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. 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. 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

5. 3
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. 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.

6. 4
1.1 OBJECTIVE OF THE STUDY
 To check the effect of the root of T. superba base diet on the fecundity of a
gravid female C. gariepinus.
 To check the effect of T. superba base diet on the latency period of a gravid
female C. gariepinus.

7. 5
CHAPTER TWO
2.1 LITERATURE REVIEW
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 and pests. Farming also involves individual or corporate ownership
of the stock being cultivated”.
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 .

8. 6
The use of medicinal plant all over the world predict the introduction of antibiotic
and other modern drugs, medicinal plants are plants that have chemoprotective and
or therapeutic effect on ailment . The medicinal properties of plants are mainly
attributes to the presence of Flavonoids, 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. 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.
In relation with different concentration of Garcinia kola in the feed of the animal’s
e.g C. 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 C. gariepinus shows increase in the egg size
(diameter).This shows that the ethanolic extracts of G. 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 G. kola seeds between 0.25 and 0.59kg feed
can be tolerated by C. gariepinus and may be included in the diets for fertility

9. 7
enhancement because this will minimize the total dependence on synthetic drugs as
fertility enhances (Dada and Ajilore, 2009). The presence of ethanolic extract of G.
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 G. kola
for the period of 8 weeks. (Adesanya et al, 2007).
Other study in man, has shown that G. 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)
In a study carried out by Majumlar et.al (1995) Albino rats of either sexes
weighing 120-150g were feed on standard diet and water and libitum. The animals
were housed at worm temp (24±2oC) on a reversed day-night cycle (06.00hrs to
18.00hrs). 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

10. 8
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 et at, 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 and Dixit, 2007).
Male mice of 8 weeks old weighing 28-32g after proper quarantine services were
gauged with 0, 0.5, 5, and 50 mg/30g body weight /day of Moringa oleifera lam

11. 9
leaves hexane fraction for 21 days with tap water used. The administration of M.
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. M.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 M. oleifera 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
M. 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 M. 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 M. oleifera has reproductive effect on the male mice.
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

12. 10
increase in the sperm count in male mice when 1% concentration of Moringa
ethanol leaf extract was administered subcutaneous for two weeks.
T. 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).
T. 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.

13. 11
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.
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.
Geographic distribution

14. 12
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 – 1000 m, Mean annual temperature: 20 – 28oC, Mean annual
rainfall: 1000 – 1800 (3000) 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 d’Ivoire 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

15. 13
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.
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.

16. 14
Plate 1. Terminalia superba plant
Source: Google images

17. 15
CHAPTER THREE
3.0 MATERIALS AND METHODS
3.1 EXPERIMENTAL SITE
The experiment was carried out at the hatchery unit of a private fish farm at
Ughelli. The experiment consisted of four treatments with each representing
different inclusion level of T. superba meal. There were three replicates per
treatment. The graded level of T. superba used were 0.00 mg/kg feed, 50 g/kg feed,
100 g/kg feed and 150 g/kg feed in the treatment respectively designated as diets
D1 to D4.
3.2 PHYSICO CHEMICAL PARAMETERS
Temperature (°C)
Surface water temperature of the pond can be determined using a mercury-in-glass
thermometer with the range of 0 to 100°C. The thermometer was lowered into the
water for about 5 minutes and the reading was taken and recorded in degree
centigrade (°C)
Hydrogen Ion Concentration (pH)

18. 16
The pH of the sample was determined by electronic method using a standard pH
meter. The electrode response was checked occasionally measuring a standard
solution of different pH, before the pH of the water sample was tested. The
electrode was thoroughly washed with distilled water and then with the sample.
The system was allowed to sterilize before the final reading was taken.
Dissolved Oxygen (DO)
The dissolved Oxygen was determined using Winkler’s method. In this method,
the water sample was collected from the fish pond in a 250 ml stopper bottle which
was immersed in the water and adhered to get filled, and then cover under the
water to ensure that of collection by addition of 2ml of Winkler A and B reagent
using a pipette. The stopper was replaced and content were thoroughly mixed. A
whitish brown precipitate which settled at the bottom was observed. 2ml of conc.
H2SO4 was to dissolve the precipitate, leaving behind a golden brown solution used
for the titration. 100ml of the sample was taken and titrated against 0.025(N)
sodium thiosulphate (Na2SO3). A pale yellow colour was formed, 2 drops of starch
solution (indicator) was added to the sample. A dark blue colouration was formed
and is then titrated against Na2SO3 until the sample become colourless at the
endpoint. The titration was done 3 times and the average titre value was
determined as
T1+T2+T3

19. 17
3

20. 18
PLANT MATERIALS AND TREATMENT
The plant materials, T. superba root were collected from Old Oyo National Park,
Oyo State, Nigeria. The roots were washed, sundried and grounded into fine
powder. Fish feeds were prepared by mixing the powder with basal feed of 40 %
crude protein, based on the formulation defined for African Catfish by Fagbenro
and Adebayo (2005) four treatment based on the mixture were used as follows 50,
100, and 150 g/kg of the powder in the basal feed designated as diets
D1(control)D2(50g/kg), D3(100g/kg) and D4(150g/kg).
3.2.1 FEED FORMULATION
Four diets were formulated from practical ingredients where the control basal diet
without T. superba roots meal and the other diets supplemented by 50, 100, and
150 g/kg of T. superba root meal respectively. The experimental diets were
formulated to contain almost 40 % crude protein. All dietary ingredients were
weighed with an electronic weigh. The ingredients were ground to a small particle
size. Ingredients including premix and T. superba roots meal were thoroughly
mixed in a pelleting and mixing machine to obtain a homogenous mass, cassava
starch was added as a binder. The resultant mash was then pressed without steam
through a mixer with 0.9mm die pore size attached to the pelleting machine. The

21. 19
produced pellets were dried at room temperature and kept frozen until the
beginning of the experiment.
Table1. Percentageoffeed 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

22. 20

23. 21
3.3 PHYTOCHEMICAL SCREENING OF TERMINALIA SUPERBA
(ROOT)
Fresh roots of T. superba collected were taken to the faculty of science central
laboratory, Delta State University, Abraka 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 Fehling’s solution was added

24. 22
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 broodfish were bought from Onosakevwe Agro
Ventures Ughelli, Delta State. The fishes had been subjected to four different diets

25. 23
with three replicate each. The broodfish were subjected to four experimental diets
at 3% body weight daily in a concrete pond for eight weeks. Diet one (D2), D3,
and D4 contains 50, 100, and 150 g/kg feed respectively while D1 which serves as
the control experiment contains 0.00 mg/kg feed. At the end of the eight weeks of
conditioning, they were subjected to artificial propagation to check the effect of the
plant on the latency period and the fecundity of the fish. The average weight of the
broodfish was 1.2±0.8kg.
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) broodfish. 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).

26. 24
3.4.2 Injection of Female broodfish
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 1 liter 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 D1, D2, D3 and D4, 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.

27. 25
The eggs were incubated using a fibre glass tank of the dimension 2×1.2×0.5m3,
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.
3.6 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).

28. 26
CHAPTER FOUR
4.0 RESULTS
The result of the physico-chemical parameters that was determined from the
broodfish pond are shown in Table1. Water Temperature ranged between 26.40-
28.40oC with a mean of 27.40oC, pH value ranged from 6.99 – 7.20 with a mean
pH of 7.10 and Dissolved oxygen ranged from 6.10 – 10.80mg/l with a mean of
8.45 mg/l during the study period.
The results of the photochemical screening of T.suparbaroot are shown in Table 2.
Where Alkaloids 2.12, Tannin 3.25, Phenol 6.35, Glycoside 1.12, Flavonids 2.64,
Steriods 2.33, Pholatannin 1.81, Saponin 10.34, Anthraquinones 3.56.
FISH AND SPAWNING
The effect of T.superba root was seen to reduce the latency period in the
experimental fish (D2, D3, and D4) to 4hours 30 minutes instead of the 10 –
12hours latency period seen in controlled fish (D1) and also it reduces the
fecundity counts of the treated fish as seen in Table 4 and 5 below.
The result of the latency period of the female broodfish is shown in Table 3.
Where9:45_10:45 is the time of injection, 2:30_3pm is the time for stripping and
the latency period lasted for 4hours 3ominutes. The result of the fecundity counts

29. 27
are represented in Table 5 with control asD1, D2, D3 and D4as the % of T.superba
root concentration in the feed, and the following result were obtained.

30. 28
Table1. Physico ChemicalParameters ofthe Water
PARAMETER Minimum Maximum Mean±S.D
Temperature oC 26.40 28.40 27.4±1.0
pH 6.99 7.20 7.10±0.11
Dissolved Oxygen (DO) mg/l 6.1 10.80 8.45±2.35

31. 29
Table 2. Photochemical Screening of T. superba.
Phytochemicals
T. superba (Root)
Inference
Alkaloids +ve
Tannin +ve
Phenol +ve
Glycoside +ve
Flavonoids +ve
Steroids +ve
Phlobatannin +ve
Saponin +ve
Anthraquinones +ve
+ve = Positive

32. 30
Table 3 Broodfish parameters
Species Weight(kg) Length(cm)
C.gariepinus 2.0-2.5kg 28.5-29.3
Mean 2.25kg 28.9

33. 31

34. 32
Table 4 fecundity counts
Treatments
Replicates
Total MeanI II III
D1 1410 1620 1457 4487 1496
D2 2016 1404 2583 6003 2001
D3 2470 1511 1443 5424 1808
D4 2270 2920 1805 6995 2331

35. 33
CHAPTER FIVE
5.0 DISCUSSION AND CONCLUSION
5.1 DISCUSSION
The phytochemical study carried out on the root of T. superba revealed the
presence of medicinally active constituents like Alkaloids, Flavonoids, saponin,
tannins, phenol, glycoside, steroid, phlobatannin, and anthraquinones. Various
phytosteroids have been shown to promote fertility (Ruiz-Luna et al., 2005).
However, the type of steroid present in T. superba has not been evaluated but this
may have a contributory effect on its profertility properties. Also, flavonoids
present in this plant have been shown to possess many pharmacological properties
such as: anti-oxidant activities, anti-inflammatory activities, anti-cancer activities
and anti-microbial effects hence, flavonoids also may have a contributory effect on
its pro-fertility properties and other pharmacological effects that the plant
possesses (Uche and Obianime, 2008; Uche, et al., 2008; Okwu and Josiah, 2006).
The results of this study on the use of T. superba root meal affects the fecundity
and latency period of C. gariepinus.

36. 34
The data obtained from the fecundity counts shows that the mean fecundity of the
female brood fish fed diets D2, D3, and D4 (2001, 1808, and 2331) was
significantly higher (p < 0.05) than what was obtained from the ovary of fish fed
the control diet D1.
A significant increase was observed in the mean fecundity count as the diet
increases against the control. This may suggest that the effect of T. superba root
meal on the fecundity counts of C. gariepinus was being enhanced after the
treatment of different dosages of the root meal. The increase in mean fecundity of
C. gariepinus in this study could be attributed to the presence of flavonoid and
xanthone in the plant. These compounds are potent anti-oxidants which are capable
of increasing the production of estrogen, the key hormone involved in the
production and maturation of eggs in the ovary.
Dada and Ajilore (2009) recorded an increase in the fecundity and egg size of C.
gariepinus fed on ethanol extract of G. kola seed diets at different inclusion level..
Adesanya et al, (2007) reported an increase in the sperm count of wistar rats after
treatment with ethanol extract of G. kola seed for 6 weeks. In another study by
Adewumi et al., (2005) they reported that C. gariepinus broodstock fed differently
with heated soybean based diets had smaller eggs and produced lower hatching
rates on fish meal-based diet.

37. 35
The result of this study therefore reveals that the root of the plant is very useful as
fecundity enhancer in C. gariepinus broodstocks management since all the
fecundity variables in this present study increased with increasing level of the T.
superba root that is dose-dependent increase.
Although the egg sizes were not determined in this study, it has however been
reported by Sule and Adikwu (2004) that species of the genus Clarias with larger
eggs also have a higher viability and endurance to starvation than those with
smaller eggs and that larger female catfish produce larger eggs.
The latency period in this study which is the difference between the time of
stripping and time of injection was 4 hours 30 minutes as against the ideal and
normal latency period of between 8 and 12 hours for C. gariepinus when using a
synthetic hormone which is also dependent on the temperature. The 4hours
30minutes latency period therefore reveals a positive effect in terms of the
reduction in the time (latency) of T. superba root on the broodstock.
The results obtained in the dissolved oxygen, pH, and temperature in this study
were within the acceptable range recommended for catfishes (Viveen et al., 1985).

38. 36
5.2 CONCLUSION
The result shows that the root of T. superba induced breeding on gravid female of
C. gariepinus.
5.3 RECOMMENDATIONS
It is therefore recommended that T. superba root can be used to induce breeding on
gravid female of C. gariepinus, instead of relying on synthetic hormones which
may be unavailable in many Africa developing countries.

39. 37
REFERENCES
Abioye, A.I.R, Duru, F.I.O., Noronha, C.C. and Okanlawon, A.O. (2003). Aqueous
extract of the bark of Kigelia africana reversis early testicular damage
induced by methanol extract of Carica papaye. Nig.J.Healht and Biomed.
Adedeji, O.S., Farimi, G.O., Ameen, S.A. and Olayemi, J.B. (2006). Effects of
bitter kola (Garcinia kola) as growth promoter in Broiler Chicks from day
old to four weeks old. J.Anim. Vet. Adv., 5(3): 191-193.
Adesanya O.A, Oluyemi K.A, Omotuyi I.O, Saalu C.L, Josiah SJ (2007).
Erythropoietic and Anti- Obesity effects of Garcinia cambogia ( Bitter
Kola) in Wistar rats. Biotechnol. Appl. Biochem. 46: 69 – 72.
Adewumi, A.A., Olaleye, V.F. and Adesulu, E.A. (2005). Egg and Sperm Quality
of the African catfish, Clarias gariepinus (Burchel) Broodstock Fed
Differently Heated Soybean- based Diets. Res J Agric Biol Sci 1(1) : 17-
22.
Akpantah A.O., Oremosu A.A., Noronhna C.C., Ekanem J.B., and Okanlawon
A.O., (2005). Effect of Garcinia kola seed extracts on ovulation, Oestrous
cycle and foetal development in cyclic female Sprague dawley rat.

40. 38
Atanda, A.N. 2007. Freshwater fish seed resources in Nigeria, pp. 361-380. In:
M.G.
Ayinla, O.A. and Akande, G.R. (1988). Growth response of Clarias gariepinus
(Burchell 1822) on silage-based diets
Billard, R.,Cosson, J., Crim, L.M. and Suquet, M. (1995). Sperm physiology and
quality. In: Broodstock Management and Egg and Larval Quality, (eds:
Bromage, N.and Roberts, J.), Black
Cabacumgan (2008). Inquirer.net, Legarda pushes for malunggay, her beauty soup.
Serrano
Chauhan N.S and Dixit V.K. (2007). Antihyperglycemic activity of the ethanolic
extract of Curculigo orchioides Gaertn. Phcog Mag. 3(12):237-240.
Dada, A.A. and Ajilore, V.O. (2009). Use of ethanol extracts of Garcinia Kola as
fertility enhancer in female catfish Clarias gariepinus broodstock. Int J
Fisheries and Aquaculture 1 (1):1-5.
Fagbenro, O.A. and Adebayo, O.T. (2005). A review of the animal and aquafeed
industries in Nigeria. In: A synthesis of the formulated animal and industry
in sub-Saharan Africa. Pp. 25 – 36. (Moel, J. and Halwart M editors).
CIFA Occasional paper No. 26, FAO, Rome, 61pp.

41. 39
Food and agricultural organization (FAO) 1998 the state of food and agriculture
table of content retrieved from www.fao.org/docrep/.../w9500e00.htm
Food and agricultural organization (FAO) of the united nations Rome (2000)
retrieved from www.fao.org/x4400e00.htm
.Froese, Rainer, and Daniel Pauly, eds. (2007). Species of Clarias in Fish Base.
Aug 2007 version.
Majumdar S.S, Tsuruta J, Griswold M.D, Bartke A. 1995.Isolation and Culture of
Sertoli Cells from the Testes of Adult
Okwu D.E. and Josiah C. (2006): Evaluation of chemical composition of two
Nigerian medicinal plants. Afr J. Biotech. 5(4): 357 – 361.
Oluyemi K.A, Jimoh O.R, Josiah S.J, Olamide AA, Omotuyi IO, Oyesola TO
(2007). Effects of Crude Ethanolic Extracts of Garcinia Cambogia on The
Owolabi, O.J. and Omogbai, E.K.I. (2007). Analgesic and anti-
inflammatory activities of the ethanolic stem barkextract of Kigelia
africana. Afr. J.Biotechnol., 6 (5): 582-585.
Pocsidio (2008). Effects of Moringa oleifera Lam Leaf Extract on Reproductive
Organ Morphology and Sperm Parameters in Male Mice. Unpub.

42. 40
Ruiz-Luna A.C, Salazar S, Aspajo N.J, Rubio J, Gasco M, Gonzales GF (2005):
Lepidium meyenii (Maca) increases litter size in normal adult female mice.
Reprod Biol Endocrinol, 3:16..
Sule, O.D and Adikwu, I.A (2004). Effect of broodstock size on egg and the
African catfish, Clarias gariepinus under laboratory conditions. J Aquatic
Sci, 19(1): 1 – 4. .
Uche F.I ,and Obienime A.W. (2008): The effects of Vitamin E on Vanadium
Pentoxide toxicity of the biochemical parameters of male guinea pigs. J.
Applied Sc & Env Mgt. 12(3): 101 – 5.
Uche F.I, Obienime A.W, Gogo – Abite M. (2008): Effects of Vanadium
Pentoxide toxicity of the histological and sperm parameters of male guinea
pigs. J. Applied Sc & Env Mgt. 12(3): 107 – 15.
Viveen, W.J.A.R., Richter,C.J.J., van Oordt,P.G.W.J., Janssen, J.A.L. and
Huisman,E.A. (1986). Practical Manual for the culture of the African
catfish (Clarias gariepinnus). Ministry of foreign affairs of the
Netherlands, Agricultural University of Wageningen and University of
Utrecht, Hague. pp 93.