CHAPTER ONE

INTRODUCTION

1.1 BACKGROUND OF THE STUDY

Fish is an important food source to man and due to the rapid increase in the population of the
world, the demand for fish and fish products as the cheapest source of animal protein have been
on the increase (FAO, 2000). Aquaculture is playing a crucial role not only in meeting the
protein need of a significant proportion of the world population but, also in boosting the
economy of most developing nations. Aquaculture techniques were developed in an attempt to
bridge the gap between fish demand and fish supply. Aquaculture has remained the most
sustainable source of fish production and supply (Abunum et al., 2021).

The main challenge faced by the Nigerian Aquaculture industry is the high cost of feed
ingredients which constitutes about 60% of the total production. Indirect devaluation of the naira
has also resulted in the hike of raw material prices, particularly fishmeal, maize, and soybean.
These have resulted in a sudden jump in the prices of aquafeeds (Abunum et al., 2021).

Commercial AquaFeeds are feeds that are formulated to meet the nutritional requirements of a
distinctive fish species and their different life stages. Two forms of commercial aquafeeds have
been identified viz. locally manufactured (mostly sinking pellets) and imported commercial feeds
(mostly floating pellets) (Abunum et al., 2021).

Despite the remarkable growth in aquaculture, many fish farms continue to stop production due
to the non-profitability of the operations as a result of the high cost of inputs especially feeds.

Like any other economic activity in life, balancing investment cost and yield is very important as
this will ensure positive returns to the farmer. It is always useful for fish farmers to know the
profitability of fish culture to guide their decision-making (Olurin et al., 2012).

Fish farmers can make more profit when they are aware of the possible ways to minimize costs
without affecting the growth of the fish. This can be achieved through proper feeding of the fish.

Catfishes of the family Clarridae comprise the most commonly cultivated fishes in Nigeria. The
growth of aquaculture in Nigeria is largely being boosted by a steady rise in catfish culture.
Since the culture of Clarias gariepinus through hypophysation was initiated in Western Nigeria
in 1973, the procedure has been practiced throughout Nigeria thus leading to increase of farm-
raised catfishes from the 80’s to date. The favoured catfishes’ species in Nigeria aquaculture
include: Clarias gariepinus, Heterobranchus bidorsalls and hybrid (Heteroclarias) and Clarias
migrodigitattis (Beingana et al., 2012).

Heterobranchus species is the more commonly cultured fish in South Eastern Nigeria. Despite
the popularity of the catfish and great market potentials, the production is still basically of
subsistence level due majorly to inadequate availability of seed for stocking and feed problems.
In Europe, about 70% of Clarias fingerling demands are supplied by producers, while in Nigeria,
however, the fingerlings supplied from both Government and privately owned hatcheries are not
enough to meet the fingerings demands (FAQ, 2010).

Nigeria is blessed with a wide diversity of inland and coastal aquatic resources including rivers,
flood plains, lakes, reservoirs, rice fields, estuaries, lagoons, mangroves and mudflats which
provide opportunities for integration of aquaculture or some form of aquatic animal management
into rural and developed areas. In Nigeria at present, sources of animal protein such as beef and
poultry come the domesticated animals that are often subjected to natural disasters. Wild animals
are almost extinct except for those conserved within game reserves (Beingana, et al., 2016).

On the other hand, over 95% of fish protein consumed in Nigeria comes from the wild with little
or no natural disasters except those caused by spoilage through poor handling and preservation
techniques (Beingana, et al., 2016).

Aquaculture in Nigeria occurs mainly Inland and only recently has the coastal region been the
focused of development. Nigeria has coast line of about 960km bordering a coastal zone which is
an extensive mangrove ecosystem comprising of lagoons, estuaries, wetlands and series of inter
connecting creeks. The coastal zone covers an estimated one (1) million hectares and offers a
considerable potential for aquaculture (FAO, 2006).

Nigeria over the years has relied heavily on fish importation to supplement her local supply
(Baruah et al., 2008). Aquaculture provides food of high quality animal protein, generates
income and employment thereby promoting the socio-economic development of Nigerians
(Baruah et al., 2008).
The culture of clariid catfish in Nigeria Clarias gariepinus (Baruah et al., 2008) family Claridae
is generally considered be one of the most important tropical cattish species for aquaculture in
West Africa (Mahmoud et al., 2019). Other species include; Heterobranchus, and their hybrids.
The reasons for their culture are based on their fast growth rate, disease resistance high stocking
density, aerial respiration, high feed conversion efficiency among others. Inorder to ensure that
crosses between Clarais gariepinus and Heterobranchus longifilis survive during culture, it is
pertinent to ensure that the proper feeds are administered.

1.2 Justification of the Study

The need to ensure that food and meat is on everyone’s table around the world is very pertinent
and imperative making it necessary to seek ways of increasing food and fish production thereby
solving the food security challenges facing Nigeria and the world at large. Breeding of fish has
several challenges such as selecting the right choice of feeds to be administered during the
process of breeding. This however becomes a big challenge as a wrong choice of feeds may be
fatal and cause diseases and sometimes death, making it compulsory to do an efficacy test of
feeds to ensure that the right choices of feeds are always administered.

1.3 Objectives of the Study

The main objective of the study is to examine the efficiency of feed utilization by offspring of
crosses between Clarias gariepinus and Heterobranchus longifilis.

1.3.1 Specific Objectives of the Study

Specifically the study seeks to

i. Obtain feeds of different kinds from reputable feeds shops within Makurdi Metropolis

ii. Obtain offspring of Clarias gariepinus and Heterobranchus longifilis from reputable
hatchery.

iii. Test the proximate content of the fish after feeding

iv. Check for water quality after feeds administration

LITERATURE REVIEW

2.1 General Biology of the Catfish

The African sharp tooth catfish (Clarias gariepinus) is a remarkable beast. It has a phenomenal
natural distribution (700 latitude), from the Cape Province in South Africa into Asia-minor
(Levic, 2008). This air breathing Clariid species exists in diverse environments; ranging from the
temperate to the tropical, and is represented in a correspondingly diverse array of aquatic faunal
assemblages, from the species poor Orange river system to species rich lake Malawi. Natural
history studies, particularly the early studies by Greenwood in the mid-1950s and later those of
Bruton in the late 1970s, have shown C. gariepinus to be a most fascinating, extremely hardy
and adaptable animal, efficiently able to exploit a wide variety of both animal and plant protein,
under diverse environmental conditions. Furthermore, it is able to withstand adverse
environmental conditions and habitat instability (Levic, 2008).

Spawning usually takes place after rain with rising water levels. Hatching, although temperature
dependent, occurs approximately 18-24 hours after fertilization at 24-28 °C (Levic, 2008).

The offspring, by exploiting newly inundated environments which are usually rich in food,
operate temporarily in an ecological vacuum, almost free from inter-and in transpacific
interactions. However, competing for food and cover which results in sibling cannibalism has
recently been shown to be density dependent under culture conditions. Growth under natural
conditions and particularly under controlled aquaculture conditions is fast. The species in an
opportunistic omnivore capable switching feeding modes, depending on prey availability
(Ghasemzadeh et al., 2010) this is principally a consequence of its wide environmental
tolerances. This ubiquitous fish is the most important individual species in traditional African
freshwater fisheries being estimated to comprise some 20% of the total catch in Africa. (Ekanem
et al., 2012). Recent revisions of the systematic of African Clarias have resulted in several
widespread species being synonynised under the name Clarias gariepinus (Ekanem et al., 2012).

These include C. capensis of southern Africa. C mossambicus of central Africa and lazera of
west and north Africa and Asia Minor. C. gariepinus has been placed in the subgenus C/arias
(C/arias) together with the vest African Species C. anguilaris, C. senegalensis and others.
Another southern African species, C. ngamensis is placed in the subgenus Clarias
(Dinotopteroides), while Heterobranchus longifilis has been placed in its own genus (Beingana,
et al., 2016). The distinguishing characteristics of Clarias gariepinus are: Head large and bony
with small eyes. Dorsal and anal fins long. No adipose fin. Pectoral fin with stout serrated spine,
used for defence or “walking” overland. Mouth terminal, large, four pairs of barbells present.
Colour varies from sandy-yellow through gray to olive with dark greenish-brown markings, belly
white. Well-developed suprabranchial organ present. The principal feature which distinguishes
Heterobranchus longifilis from Clarias gariepinus is its prominent adipose fin (Shipron and
Hasan, 2013).

2.2 Production of hybrid catfishes

The mating or crossing of two different species is a process called hybridization, with the
offspring known as hybrids. Hybrids can have some characteristics of both parents. Breeding
hybrid with selected or favored characteristics of both parents is one of the goals of animals’
husbandry (Beingana, et al., 2016). When a hybrid has characteristics superior to both parents it
is said to have hybrid vigor or positive heterosis which, of course, is the ultimate breeding goat
(Beingana, et al., 2016). Hybrids between different species of North American catfish
(Ictalurids) have been researched for more than 30 years. Of all these interspecific catfish
hybrids (crosses between two distinct species) only one hybrid has characteristics that would
favour commercial application. That hybrid is the channel catfish (Ictalurus punctatus) blue
catfish. (I. furcatus) hybrid (denoted as the C x B (hybrid). More specifically, it is the hybrid
produced by crossing the female channel catfish with the male blue catfish. It is important to
note that the reciprocal cross, crossing the male channel catfish with the female blue catfish, does
not have the same superior production characteristics of the Channel catfish hybrid. Research on
Channel catfish hybrid hybrids has demonstrated that they exhibit many commercially desirable
characteristics. The Channel catfish hybrid exhibits superior or characteristics for the following
traits.

• Faster growth;

• better feed conversion;

• Tolerance of low oxygen;

• Tolerance to crowded growth conditions in ponds;

• Uniformity in size and shape;

• Higher dress out percentages

• Increased harvest by seining; and

• Increased vulnerability to angling.

2.3 Monoculture

Monoculture involves the culture of a single species with or without the addition of fertilizers or
feeds. Fertilizers commonly used by local fish farmers are manures from cows, buffalo, pigs, and
chickens. Inorganic fertilizers are not commonly used. A typical example of controlled stocking
monoculture with the addition of organic fertilizers is the culture of Nile Tilapia, (Sarotherodon
niloticus), with a stocking density of 2 fish per m2 of pond surface area, production of 25000kg
per ha at least twice as many as fish as can be produced from naturally occurring organisms, is
possible after four months of rearing by adding pig manure every day for five days a week at the
rate of 75kg per ha per day. Supplementary feeding is another means that can increase the
production fish culturing in ponds. At present, some commercial fish farmers use rice bran and
broken rice to feed fish. Water spinach, duckweed, and other aquatic weeds, soybean, cake,
peanut cake, fish meal, and fresh or frozen trash fish are also used.

2.4 Culturable fish species in Nigeria

There are great varieties of fish that can grow in the ponds. All warm- water fish can grow in
ponds. But some are predominantly fresh water while some are brackish water fish. It is
observed that the freshwater fish do better in freshwater ponds and the brackish water fish grows
better in brackish water ponds although in some cases freshwater fish can be acclimatized to
grow in brackish water and vice versa (Shipton and Hasan, 2013).

2.5 Importance of Fish Feed

In general, the farming of aquatic organisms is divided into intensive, semi-intensive and
extensive depending on the degree of human intervention in their management, which is
gradually changing from extensive to intensive farming. This intervention can be quantified in a
substantial way with the intake of food from outside, the elimination of catabolites and the
supply of oxygen in the farmed water.

Knowledge of the nutritional requirements of farmed fish species, the formulation of animal
feed, and the management of animal feed on the farm in recenta decades has been the subject of
much research, though have not yet reached the level of other species of zootechnical interest.

In the diet of fish, we began decades ago to use raw foods and by-products from the processing
industry, with whole foods made from high quality raw materials. Considerable attention was
then given to the management aspects of feed distribution and environmental problems, with the
aim of minimizing wastage of organic matter, pollution and eutrophication of water.

Feeding intensively farmed aquatic organisms has evolved over the past 40 years, which has
greatly increased the productivity of aquaculture around the world. In the early, fish waste or
whole fish was used to feed the fish. However, this practice was not very effective because it
produced a large amount of waste that polluted the water considerably, making it impossible to
increase the density of farmed fish, knowing that the growing demand for fish has encouraged
intensive farming. The practice of using forage fish to feed farmed fish is also very risky from a
health point of view, as many potentially pathogenic organisms are introduced into the livestock
environment.

The next step towards optimizing fish feeding techniques has been the use of mash, made from
stable ingredients such as fishmeal, vegetable flours and fish oils. Pelleted feed followed the wet
feed, which had the advantage of being produced and stored on the farm, or it could be
purchased from feed companies, which then began to develop diets that optimized their
composition based on high species, and the size of the fish.

The latest evolution of the aquaculture diet has been the introduction of extrusion. This
production technique has brought two great benefits. The first is to be able to choose the degree
of floatation of the food, by acting on the density of the extrudate. This allows today to have
floating granules, or semi-floating. In fact, the behavior of food in the water column is a very
important aspect to support the attitudes of different species of fish to food. The second
advantage provided by the extrusion process concerns the possibility of considerably increasing
the lipid percentage of the food, thanks to its porosity. This has led to the formulation of high
energy diets containing up to 30 to 35% lipids.

The amount of fat contained in an aquaculture feed is of great importance for the use of nutrients
by high aquatic organisms. Fish, especially carnivorous species (Salmonidae), have a low
digestive capacity in carbohydrates. They prefer to use proteins and fats as a source of energy. A
diet capable of providing the energy needed to metabolize fish through lipids allows for a large
saving of proteins, which will then be used more for growth and flesh formation, instead of
energy source.

Using less protein for energy purposes is important for both: Have a cheaper diet, given the cost
of fishmeal.

The use of amino acids as a source of energy encourages the release of pus of nitrogenous
material by fish, which has a major impact on the pollution of aquatic environments.

Aquaculture is the fastest growing zootechnic sector in the world. Its annual growth rate has been
10% in recent years. The expansion of the sector has been very rapid in recent decades, leading
to an ever-increasing demand for raw materials for the production of fish feed, particularly fish-
derived meals and oils. These two ingredients come mainly from the processing of small pelagic
fish caught and not intended for direct human consumption. Only a marginal part of the fishmeal
and fish oils used in feed formulation comes from the processing of fish waste for human
consumption.

Replacing fish oils and meal with other plant-based products was also needed to address the
sustainability issue by looking at the relationship between the amount of fish used in food
formulation and the amount of high fish. This relationship called fish in fish, was very
unfavorable in the past, some fish production using up to 5 kg of fish caught to produce 1 kg of
farmed fish. Now, thanks to progress in feeding, this ratio has been reduced for all species raised,
even for strictly carnivorous species. For example, today a farmed salmon is able to grow faster,
ingesting less fish for its nutrition than a specimen of the same wild species. Thanks to the
genetic selection of farmed fish and the formulation of increasingly balanced feeds, the
aquaculture sector is able to provide extremely efficient feed conversion indices, the best of all
animals in the world. For some species, we have indices below 1, that is, less than 1 kg of food,
which contains more than 95% dry matter, is required for the growth of one kg of fish.

2.6 The Physical and Chemical Environment of Fishes

A good water source is a necessity for the survival and growth of fish since the entire life
processes of fish is solely dependent on its environment. Theses water quality parameter become
more serious in intensive culture system where fish is raised in artificial ponds with reduce self-
purification capability as pared with natural system. For example, the intensive artificial feeding
has identified as a major source of water pollution in fish ponds as only about of the nutrients in
the feeding is converted to fish flesh (Shipton and Hasan, 2013), while are remaining settles as
wastes when decomposed will utilize all the dissolved oxygen. The constant check of these water
quality parameters is necessary to ensure a conducive environment for the growing fish.
According to (Shipton and Hasan, 2013), any physico-chemical biological characteristic of water
which is the survival growth and reproduction of the fish is a water quality variable demands, the
attention and management of the ponds culturist. These quality parameters could be physical,
chemical and biological characteristics (Shipton and Hasan, 2013).

2.6.1 Temperature

This is regarded as the environmental factor that the activity, behaviour, Feeding, growth,
survival, appetite and reproduction in all fishes.

Temperature is considered the next most important water quality parameter to dissolve oxygen.
Temperature variation in water bodies depend largely on their geographical location (Latitude,
Longitude and altitude).

In the tropics surface temperature of shallow water bodies such as ponds, tanks, Lagoons can
reach near 40°C or above in peak dry season causing high mortality of fish especially if the water
bodies are less than lm depth. More often temperature of water bodies ranges from 30-35°C,
most warm water fishes tolerate and farewell in this range as reported by Boyd and Lichtkopple.
In the temperate regions, daily maximum temperature rarely reaches over 25°C. Therefore,
temperature has a pronounced effect on rate of chemical and biological processes in water, for
example animal require twice as much oxygen at 30°C than at 20°C.
Thus, excretion may be necessary at higher temperature to increase the level of dissolved
oxygen. The main cause of thermal stratification in ponds is temperature. The hypolimnion often
associated with Low primary productivity and low dissolved oxygen.

2.6.2 Dissolved oxygen

The dissolved oxygen content depends on the pond water temperature. It also depends to a
considerable degree on the quantity of organic matter present and submerged aquatic vegetation.
If decomposing organic matter is too much, it will absorb too much of the dissolved oxygen in
water, and the level will fall. If vegetation is also too dense, during the day it can give off excess
oxygen to point of saturation beyond 150% of the normal level, and this over saturation endanger
the lives of the fish, and the fry in particular. At night the same vegetation absorbs oxygen
content below minimum level acceptable to the fish. The danger is greater in warm and thundry
weather, and when there is a sundry-abundance of aquatic algae. Boyd and Lichtkopplerb
reported that in the absence of deliberate poisoning dissolved oxygen is the simple most
important and critical water quality parameter for fish in pond and culture systems. For
salmonids in general 9mg/L are necessary which corresponds to temperature of around 20°c, in
pure water, 5.5mg/L is the minimum. Cyprinids require about 6 to 7 mg/L, and a minimum of
3mg/L, Cichlids and Silurids can withstand much lower content and in consequence, high
temperature and water in organic matter, both of which reduce the dissolved oxygen content in
the aquatic environment. Shortage of dissolved oxygen has many effects on fish which include
fish coming to the surface in an effort to breath air, fish stop feeding, growth and reproduction is
impaired, fish group together near the fresh water inlet, fish become more susceptible to
diseases, may become stress and may subsequently die (Adeniyi and Ovie, 1990).

Hydrogen concentration: potential hydrogen pH

Although this depend on the species to be cultured, but in all cases a pH range of 7.0-9.0 has
been identified as productive and suitable for fish farming (Ndukwe 2006)

Fish in fresh water strive well in alkaline base environment. Regular test should be carried out to
control the acidity or alkalinity of the culture water which promotes toxicity of chemical
compounds in the water (Ndukwe, 2006). The water levels can make the effects of these
substances, as a general rule the following pH values can be used to see if there is water
pollution as depicted on

Ammonia

Ammonia is extremely toxic at low levels poses a threat to fish health. Ammonia is produced by
fish as normal part of metabolism. Ammonia is so toxic that most animals immediately convert it
to less harmful substances, usually urea. Fish shortcut this process and continually excrete
metabolic ammonia directly into the surrounding water via special cells in the gills. Therefore,
the confines of aquaria and ponds, levels can rise rapidly to dangerous levels. At low levels
(<0.lmg/L NH3) acts as a strong irritant to the gills with prolonged exposure. At higher levels
(>0.mg/L NH3) even relatively short exposure can lead to skin, eye, and gills damage (Robert
and Gene, 1997). When dissolved in water, normal ammonia (NH3) reacts to form ionized
ammonium (NH3 + H2O NH4 + OH-) Ammonia (NH3) is highly toxic, whereas the ammonium
ion is less toxic. At any point in time there will be both ammonia molecules and ammonium ions
present. The quantity of both ammonia and ammonium for each species is dependent on both pH
and temperature. As a thumb rule it is best to aim at a zero level of total ammonia at all time. In
normal circumstances any reading above 0lmg/L Total Ammonium Nitrogen (TAN) should be
considered as unacceptable (Robert and Gene, 1997).

Nitrite

Elevated levels of nitrite are common when people set up new tanks or pond. Nitrite level only
increase in presence of nitrifying bacteria which are slow growers. Until fishes are well
established nitrite levels may be significantly low and acceptable to fish health. Also nitrite
levels are influenced by level of ammonia in the water. Nitrifying bacteria also process ammonia
to nitrite (Vijal et al., 2002). Furthermore, nitrate is converted to nitrite which is usually
considered as harmless at levels less than5omg/L. safe levels of nitrite in water r aquatic life is
0.06mg/L (Robert and Gene, 1997). National Guidelines and standard for water quality by the
Federal Ministry of Environment prescribe the following as standard for water quality for aquatic
life.
 CHAPTER THREE

MATERIALS AND METHODS

3.1 Sample Collection

114 fingerlings will be obtained from reputable hatcheries within Makurdi Metropolis and
transported to the department of Fisheries and Aquaculture Laboratory Joseph Sarwuan Tarka
University Makurdi for Further analysis.

Experimental procedure

cA total number of 144 fingerlings will be divided into four culture groups and each group was
in duplicate with each

group having 18 fingerlings. The culture groups were;

Treatment1: Monoculture of Clarias gariepinus

Treatment 2: Monoculture of Heteroclarias

Treatment 3: Monoculture of Heterobrachus longifilis

Feed formulation

All the materials used in the feed formulation were bought at at a Reputable Hatchery within
Makurdi Metropolis. The percentage crude protein composition of the feeds will be obtained
using Pearson method.

Pellteting of feeds

A locally fabricated pelleting machine with 2.00mm

diameter die will be used to produce the pellets for the fingerlings at Kiyi Fish Farm. Before the
pelleting, the ingredients were properly mixed manually and then binded with starch. This was
then placed in the funnel of the pelleting machine and switched on to produce the pellets. The
pellets will then be air dried on a clean mat outside in a shade to ensure that they were properly
dried. This will be done by weighing the pellets daily until a uniform weight will be obtained.
They will then then be packed in nylon bags to prevent mould, pests and water from entering.
Proximate analysis of fish and feeds

At the beginning of the feeding trial, five (5) fingerlings will be removed from the treatments,
weighed and oven dried at 600 to determine the moisture, protein, ash and lipid contents.
Similarly, analysis for all the feed stuffs was also cone to determine the moisture, protein, ash
and lipid content. At the end of the experiment one fish was randomly selected from each
treatment, weighed, oven dried at 60°C for 24 hours and then processed for analysis of moisture,
protein, ash and lipid contents. All these will be done at the department of Fisheries and
Aquaculture Laboratory Joseph Sarwuan Tarka University Makuridi.

Determination of moisture

Moisture will be assessed using the AOAC air oven method (1995). 2 g of the test material was
weighed in triplicate into previously weighed and cooled Petri dishes. These will be placed in an
oven and then placed in an oven where air oven at 105°C±2°C for 3 hours. The samples were
placed in the oven, covered, and allowed to cool for 5 minutes before being weighed. The plates
were reweighed in the oven until constant weights were obtained. The moisture content of
feedstuff, diets, and carcasses was determined and expressed as a percentage of the initial sample
weight.

% Moisture=W1-W2 x 100 1

W1

Where W1 = weight of fresh sample

Where W2 = weight of sample after drying

Determination of protein

The micro-kjeldahl method as described by AOAC (1995) was used for the determination of
protein in test samples.

About 200mg ground sample will Be weighed and placed in micro-kjedahl flasks followed by 1g
of catalyst and digested in 20 mls concentrated sulphuric acid. This will be heated for about 2 ½
hours on a digestion stand. The blanks containing only the sulphuric acid and catalyst will also
be heated for the same period. Distillation: About 2m1 of distilled water was added to the sample
and poured in Mark ham’s distillation apparatus followed by excess sodium hydroxide slightly
above 5ml. the liberated ammonia was distilled into 50m1 conical flask containing 2 % boric
acid to which 3 drops of methyl red indicator will be added.

Determination of ash

This measured the total in-organic matter by incineration. The procedure outlined in AOAC
(1995) will be used in determining the crude fibre content of samples. The weight of the crucible
dish will be taken. 2 grammes of sample will be added to each of the crucible.

The content will be aced on the furnace rack and the furnace temperature was set to 500°C for 16
hours until sample completely ashed. The ash in crucible will be removed and kept in desiccators
to cool. The cooled ash and dishes will be re-weighed and percentage ash will be calculated as;

Percentage ash = Total weight of extracted ash x 100 3

Weight of sample

Determination of lipid

The Soxhiet extraction method outline in AOAC (1995) was used in determining the lipid
content of the samples.

Two grammes of the samples will be weighed and the weight of the flat bottom flask was taken
with the extractor mounted on it. The thimble was held half way into the extractor and the
weighed sample will be carefully transferred into the thimble. Extraction will be plugged with
cotton wool fully dropped into the extractor and the extraction was continuous for 8 hours.

At the completion of the extraction, the solvent will be removed by evaporation of the water bath
and the remaining part in the flak dried to 80°C for 30 minutes in the air oven to dry the solvent
and cooled in a desiccator.

The flask will be reweighed and percentage lipid calculated as:

Percentage Lipid = Total weight of extracted lipid x 100 4

Weight of sample 1

Feeding
The fishes will be fed daily on formulated experimental diet containing 35% crude protein twice
while the second culture treatment will be fed with commercial feed (Coppens) as the artificial
feed at3% of their total body weight based on the recommendation of (Viveen et al., 1986). This
feeding rate is close to the apparent satiation of the fish as reported by Wilson (1989). The daily
ration will be divided into two, one portion will be fed at 09. O0hrs and the other at 18.00hrs.
Feed will be dispensed evenly on the water surface to allow for equal opportunity for feeding.
Feeding in all tanks were completed in about 10 — 15 minutes.

Collection of data (length/weight) will be carried out once a week during the experiment. Fishes
in monoculture system will be weighed one after the other and the total sum will be calculated.

Tank management

The plastic tanks were bought at Mordern market.

The diameter of each of the plastic tank will be 64cm while the height was 32cm giving a
volume of 3,218.3cm3. Prior to the introduction of the fishes, the tanks will be washed
thoroughly with salt to kill any pathogens. The tanks were then filled with the brehole water to
50 litres capacity.

Eighteen fingerlings will then be introduced to each of the tanks in the monoculture system.

Physicochemical parameters

Temperature

The temperature of water in all bowls will be monitored once a week. This was by dipping a
mercury-in-bulb thermometer into the water below the water levels in the bowls. The
thermometer stayed in the water for about 3 minutes before taking the reading. This will be
repeated thrice. Each temperature reading will be taken in degree Celsius.

pH: pH will be taken before draining the water in each treatment tank. This will be done
effectively using pH meter model no. H3351C-Kentel 704546

Dissolved oxygen

Once a week, water samples from the tanks were collected for dissolved oxygen (DO) analysis.
The Winkler Titrimetric technique was utilized (Annes, 1966).
The steps will be as follows: 2m1 of Manganese alkaline iodide reagent (125-300ml) in the
sampling bottle by inverting the bottle a few times. When the settled precipitate will be added,
the stopper was gently removed, and 2.0ml concentrated sulphuric acid (H2SO4) was added by
letting the acid to trickle down the neck of the bottle — stopper, and mixing by gentle inversion
until the solution was completely formed. This was titrated with 0.025N sodium thiosulphate
(Na2S2O2) to a pale straw colour. 3 drops of freshly prepared starch solution will be and the
titration continued till the first disappearance of the blue colour, the total volume of Na2S2O2
used. The dissolved oxygen (DO) in mg/1 will be calculated as:

DO mg/l = (MiI.titrant) (N) x (8) x (1000) 5

Sample Volume

Where N = Normality of Na2S2O3

8= Concentration equivalent to 1ml in Na2S2O3

1000 =Conversion factor for 1 litre

All readings were in mg/1 of water.

Ammonia

To determine the ammonia, l0ml of the water sample will be taken and 0.2m1 Zinc Sulphate
(ZnSO4) added.

0.4ml sodium hydroxide (NaOH) was added and mixed thoroughly. It was allowed to stand for at
least 15 minutes and centrifuged for 15 minutes. 2mls aliquots were taken and to it Rochelle salt
was added. This was shaken and 0.5m1 Nessler’s reagent was then added. It was allowed to
stand for 20 mm. The ammonia reading was taken in mg/l.

Nitrite

The procedure for Nitrite determination was as follows:

The water sample was filtered through Watchman ‘42. Then, 50ml of the water was measured
into a 100 ml beaker. 1ml of diazotizing reagent (sulphanilamide) was added, stirred and allowed
to stay for four minutes for reaction. 1ml of coupling reagent (N-CI-napthyl-ethyleneamine) was
added and stirred. The solution was allowed to stand for 10 minutes to form the ozo compound
and then transferred to a 1cm cuvette where the pink colour was measured
spectrophotometrically at 543nm the concentration of Nitrite Niitrogen was obtained from a
calibration graph.

Analysis of growth and nutrient utilization

Growth and nutrient utilization parameters will be analysed using the following formulae as
reported by Anadu et al. 986)

1. Weight gain (WTG) = Final mean weight - initial mean

weight 6

2. Specific growth rate (SGR)

SGR (% per day) = Lnw2 — Lnw1

Culture period 7

Where W1= initial weight (g)

W2 = final weight (g)

Where Ln = natural logarithm

3. Feed conversion efficiency; = final weight gain x 100 8

Protein intake 1

4. Survival rate (percentage) = No of dead harvested fish x 100

No of fish stocked

Data analysis

Data obtained from fish weights in this study will be subjected to analysis of race (ANOVA)
using the SAS ANOVA procedure (SAS Institute, 1988). Duncan’s multiple range test (Duncan,
1955) will be used to compare differences among treatment means 5% level of confidence.
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