THE EFFECT OF NATURAL PLANT EXTRACTS (BITER LEAF AND NEEM LEAF)

AS PROPHYLACTIC TREATMENTS ON THE GROWTH AND SURVIVAL OF Clarias

gariepinus FRY TO FINGERLINGS

BY

AUGUSTINE BENJAMIN

SOA/HND/FIT/19/016

A PROJECT REPORT SUBMITTED TO THE DEPARTMENT OF FISHERIES

TECHNOLOGY, COLLEGE OF AGRICULTURE, SCIENCE AND TECHNOLOGY,
LAFIA, NASARAWA STATE

IN PARTIAL FULFILLMENT OF THE REQUIREMENT FOR THE AWARD OF

HIGHER NATIONAL DIPLOMA (HND) IN FISHERIES TECHNOLOGY

APRIL, 2022

i
 DECLARATION

I declare that the work in this project report entitled “EFFECT OF NATURAL PLANTS

EXTRACTS (BITTER LEAF AND NEEM LEAF) AS PROPHYLACTIC TREATMENTS ON

THE GROWTH AND SURVIVAL OF Clarias gariepinus FRY TO FINGERLINGS ”

represents my original work and has not been previously submitted for award of any certified in

this or any other institution.

AUGUSTINE, Benjamin ------------------------------- ------------------

SOA/HND/FIT/19/016 Signature Date

ii
 CERTIFICATION

This project report titled “EFFECT OF NATURAL PLANTS EXTRACTS (BITTER LEAF

AND NEEM LEAF) AS PROPHYLACTIC TREATMENTS ON THE GROWTH AND

SURVIVAL OF Clarias gariepinus FRY TO FINGERLINGS” by AUGUSTINE, Benjamin

with Registration number SOA/HND/FIT/19/016 meets the regulations governing the award of

Higher National Diploma (HND) in Fisheries Technology of the College of Agriculture, Science

and Technology, Lafia, and is approved for its contribution to knowledge and literacy

presentation.

……………………………….. ………………….. ………………………

Mr. Usman Manasseh Signature Date

Project Supervisor

………………………………. ……………………. …………………

Mr. Usman Manasseh Signature Date

HOD FISHERIES TECHNOLOGY

………………………………. ……………………. …………………

Dr. S.I. Iorchor Signature Date

(External Examiner)

iii
 DEDICATION

This project is dedicated to God Almighty and to my parents Mr. and Mrs. Augustine for their

care and support throughout my education.

iv
 ACKNOWLEDGEMENT

I express my profound gratitude to Almighty God who gave me the grace up to the end of my

research work.

I acknowledge the effort of my supervisor, Mr. Usman Manasseh for devoting his time and

energy to see the success of this work. It is my prayer that God Almighty will grant you the

wisdom, knowledge and strength in all your endeavour.

I also acknowledge my Head of Department, Mr. Usman Manasseh and the entire staff of

Fisheries Technology Department for their contribution in one way or the other to the success of

this work.

I heartily appreciate the enormous moral, spiritual and financial support of my parents, Mr. and

Mrs. Augustine towards my life entirely. May the good Lord reward you all. Amen.

v
 TABLE OF CONTENTS

Content Page

Title page - - - - - - - - - - i
Declaration - - - - - - - - - - ii
Approval page- - - - - - - - - - iii
Dedication - - - - - - - - - - iv
Acknowledgement - - - - - - - - - v
Table of contents - - - - - - - - - vi
List of tables - - - - - - - - - - viii
Abstract - - - - - - - - - - ix
CHAPTER ONE

1.0 INTRODUCTION - - - - - - - - 1

1.1 Background of the study - - - - - - - 1

1.2 Justification of the study - - - - - - - 5

1.3 Aim and objectives of the study - - - - - - 5

CHAPTER TWO

2.0 LITERATURE REVIEW- - - - - - - - 7

2.1 Use of medicinal plants as Immunostimulants in Aquaculture-- -- - 7

2.2 Plants used as Immunostimulants in Aquaculture- - - - - 8

2.3 Administration and Application Methods of Immunostimulants - - - 11

2.4 Mode of Action of Immunostimulants - - - - - - 14

2.4.1 Improve the Innate Immune Responses- - - - - - 14

2.4.2 Enhance Antimicrobial Activity- - - - - - - 15

2.4.3 Herbal Plants as Promising Antibiotics- - - - - - 16

vi
2.4.4 Enhance the fish growth, Feed Utilization and Nutrient Digestibility - 17

2.6 Neem-Azadirachta indica- - - - - - - - 18

2.6.1 Neem-Azadirachta indica usage in aquaculture- - - - - 19

CHAPTER THREE

3.0 Materials and methods - - - - - - - 20

3.1 Location of the study - - - - - - - - 20

3.2 Procurement of experimental fish - - -- - - - 20

3.3 Collection and processing of Plant extracts bitter leaf and neem leaves- -- 20

3.4 Experimental set up - - - - - - - - 21

3.5 Data Collection - - - - - - - - 21

3.6 Statistical Analysis - - - - - - - - 22

CHAPTER FOUR

4.0 Results and Discussion - - - - - - - 23

4.1 Results- - - - - - - - - - 23

4.2 Discussion - - - - - - - - - 28

CHAPTER FIVE

5.0 Summary, Conclusion and Recommendation- - - - - 32

5.1 Summary - - - - - - - - - 32

5.2 Conclusion - - - - - - - - - 32

5.3 Recommendation - - - - - - - - 32

References - - - - - - - - - 34

vii
 LIST OF TABLES

Table 1: Growth performance of Clarias gariepinus fry treated with bitter leaf and neem leaf
extract - - - - - - - - 24

Table 2: Disease Resistance of Clarias gariepinus fry treated with bitter leaf and neem leaf
extract - - - - - - - - 25

Table 3: Water Quality Parameters of Clarias gariepinus fry exposed to bitter leaf and neem leaf
extract - - - - - - - - - 27

viii
 Abstract

Growth Performance and Survival rate of Clarias gariepinus Fry Fed Neem leave and bitter
extracts as prophylactic treatments was carried out at College of Agriculture, Science and
Technology, Lafia, Department of Fisheries Technology.5000 five day old fry of Clarias
gariepinus fry were sourced from the hatchery unit of the fish farm, they were acclimatized for a
period of seven days and were feed 3 times daily. The experiment was carried out in four
treatment three replicate, bitter leaf and neem leave for a period of 12 weeks. The acclimatized
fry will be selected randomly and stocked at 50 fry per bowl containing 15 litres of bore-hole
water. Biometric parameters like body weight of fish were measured using digital weight balance
(mettle Toledo AB54) respectively, feeding was done 3 times daily at 3% body weight. Flow
through system was ensured throughout the period of the experiments. Sampling was done from
the beginning to the end to determine the growth rate of the fry. Water quality analysis of the
source of water was monitored and the result was within the acceptable range, they were not
significantly different from each other. Neem leave and bitter leaf extract improve the growth
and survival rate of Clarias gariepinus fry. The inclusion of bitter leaf at 200ml gave the best
growth performance in fish. The use of neem leave and bitter leaf extract is highly recommended
for farmers towards progressive growth and survival rate of Clarias gariepinus in aquaculture
industries.

ix
 CHAPTER ONE

1.0 INTRODUCTION

1.1 Background of the Study

As compared to the other food production sectors, the aquaculture industry has flourished

significantly in recent times to meet the requirements of the fish market. Fish are

considered to be the primary source of nutrition in most poor countries, thereby creating a

huge demand (FAO, 2018) The African catfish, Clarias gariepinus, which belongs to the

family Clariidae, is a widely cultured species in tropical and subtropical countries.

Owing to its hardy nature, high cultivability, fast growth rate, reproductive ability in

captivity, high survivorship, the capability to accept formulated feed, and somewhat

cheaper in comparison with other fish species, African catfish is one of the most

preferred fish species for culture in Africa (Adeshina & Abdel-Tawwab, 2020, Kemigabo

et al 2018). They can dwell a wide range of freshwater habitats like rivers, dams,

floodplains, swamps, and lakes. Because of their accessory air-breathing organ, they can

survive in hostile environments in turbid, muddy, and oxygen-depleted water bodies

(Dadebo, 2000, & Idodo-Umeh 2003). Being omnivorous in feeding nature, they can feed

on plankton, insects, plants, and snails in the natural water bodies (Dadebo, 2000).

The fundamental objectives of the fish production industry are to improve fish growth,

nutrient digestibility, immunity, and decrease feed cost (El-Araby et al., 2020 , Dada,

2017 & Bostock et al., 2010). The fish reared under intensive culture, or nutritionally

deficient or physiologically unbalanced environments are more susceptible to a wide

diversity of bacterial pathogens (Mokoro et al., 2014). These pathogens incur economic

1
losses in the aquaculture industry resulting from mass mortality, cost of treatment, and

reduced production (Ashiru et al., 2011). One of these opportunistic pathogens,

‘Aeromonas hydrophila, a bacterium, can infect numerous fish species inducing

hemorrhagic septicemia (Zhang et al., 2014). Aeromonas infection is usually treated with

antibiotics, often developing drug-resistant bacteria and posing a threat to human health .

Therefore, safe and cheap alternatives are essential to confront these bacterial diseases.

The use of plant-based additives in fish diets is regarded as one of these alternatives

(Dada, 2017).

Nigeria as a country consumed fish more than any other country in Africa and is among

the largest fish consumers in the world with over 1.5 million tons of fish consumed

annually. However, over 900,000 metric tons of fishes are imported annually while its

domestic catch is estimated at 450,000metric tons/year (Ozigbo et al., 2013). Federal

Department of Fisheries (2008) has reported that growth in fish production is as a result

of increased activities of aquaculture, and the need for aquaculture arose from the

decrease in supply from ocean fisheries as a result of over-fishing, habitat destruction and

pollutions (Adedeji and Okocha, 2011). Fish and fishery products represent a very

valuable source of protein and essential micronutrients for balanced nutrition and good

health. In Nigeria, fish provides 40% of the dietary intake of animal protein of the

average Nigerian. Aside from being a source of protein for livestock, fish plays an

important role medicinally as it replenishes the human body with vitamins A and D;

calcium, phosphorus and lysine; sulphur and amino acids (Ohene- Adjei et al., 2007).

Water bodies in Nigeria harbour a variety of fish species that serve as food and an

economic resource to the country. Some of the most important species that account for

2
90% of Nigeria’s fishery includes croaker, catfishes, tilapias, threadfins and the clupeids

(Federal Department of Fisheries, 2003).

Fish production in Nigeria could not meet domestic demand. The demand for fish in

Nigeria mostly outstrips the local production. Widespread homestead and small scale fish

production can substantially solve the demand–supply gap in the country. Efforts made to

improve fish production in the country must be anchored on analysis of fish production

(Kudi et al., 2008). Aquaculture has been revealed as the main source for increasing fish

supply. It is the fastest-growing animal food producing sector. World aquaculture

production may result in an increase in disease outbreaks which has been reported due to

culture intensification, resulting in partial or total loss of production (Bondad-Reantaso et

al., 2005). Factors such as poor water quality, overcrowding, periodic handling, high or

sudden changes in temperature, and poor nutritional status contribute to physiological

changes in fish such as stress or immunosuppression and thus, heighten susceptibility to

infection. Parasites and pathogens induce biological stress on fishes (Ukwa, 2012; Ukwa

et al., 2015), reducing their productivity (Cabello, 2006; Naylor et al., 2000). Lack of

sanitary barriers facilitates the spread of pathogens, producing high mortality levels

(Cabello, 2006). Like humans and other animals, fish suffer from diseases and parasites.

Disease is a prime agent affecting fish mortality, especially when fish are young. Fish

parasites cause commercial losses in both the aquaculture and fisheries industries and

may have human health, as well as socio-economic implications both in developing and

developed countries.

In order to avoid economic losses related to sanitary shortcomings, several veterinary

drugs are commonly used in aquaculture to prevent or treat disease outbreaks.

3
Antimicrobials and other veterinary drugs are administered regularly as additives in fish

food or sometimes in baths and injections and are used as prophylactics (prevent diseases

before they occur), therapeutics (treat sick animals) or growth promoters. Nevertheless,

the use of veterinary drugs is becoming more restricted since they present numerous side-

effects for the environment and health safety. For example, massive use of antibiotics

have resulted in the development of resistant bacteria strains (Mirand and Zemelman,

2002) or the presence of residual antibiotics in the muscle of commercialized fish and

thus has potential consequences on human health (Cabello, 2006). The use of drugs like

praziquantel in bath treatments for parasites has also numerous disadvantages like

development of resistance (Umezawa et al., 2006), being hazardous for animal health and

environmental disadvantages. Vaccination has also been regarded as a potential

treatment against disease outbreaks in aquaculture. However, commercial vaccines are

too expensive for widespread use by fish producers and they have the downside that a

single vaccine is effective against only one type of pathogens (Harikrishnan et al., 2011).

Considering the potential harm of veterinary drug treatments on the environment and

human health and in some cases their limited efficacy, disease management should

concentrate on harmless, preventive and lasting methods. Medicinal plants are promising

to be an important source of therapeutics in fish culture since these products provides a

cheaper source for treatment, eco-friendly and greater accuracy without causing toxicity

(Madhuri et al., 2012). Plants are rich in a wide variety of secondary metabolites of

phytochemical constituents such as tannins, terpenoids, alkaloids and flavonoids, which

act against different diseases (Pandey and Madhuri, 2010; Ravikumar et al., 2010).

Phytochemicals are known to possess antioxidant, antibacterial, antifungal, antidiabetic,

4
 anti-inflammatory, antiarthritic, and radio-protective activity, and due to these properties

they are largely used for medicinal purposes (Nair et al., 2005). Medicinal plants could

have a more promising role in the future as drug target for the control of helminth

infections in the tropics (Hammond et al., 1997). The present study was conducted to

determine the effect of natural plant extract biter leaf (Vernonia Amygdalina) Neem leaf

(Azadirachta indica) as prophylactic treatments on the growth and survival of Clarias

gariepinus fry to fingerlings.

1.2 Justification of the study

Antibiotics and anthelminthics are frequently used to control disease caused by these

parasites, but the continuous use of these synthetic agents in aquaculture have resulted in

more resistant parasitic strains. The prophylactic use of medicinal plants has gained

considerable momentum in the world during the past decade. The over use of synthetic

drugs with impurities, resulting in higher incidence of adverse reaction, has motivated

mankind to go back to nature for safer remedies.

1.3 Aim and Objectives of the Study

The aim of the study is to determine the effect of natural plant extracts, (bitter leaf, neem

leaves) as prophylactic treatments on the growth and survival of Clarias gariepinus fry

to fingerlings.

The objectives of the study are:

i. To determine the effect of bitter leaf and neem leaves on the growth and survival of

Clarias gariepinus fry to fingerlings.

5
ii. To determine the prophylactic properties of natural plant extracts (Vernonia amygdalina,

Azadirachta indica ) on the growth and survival of fry to fingerlings.

6
 CHAPTER TWO

1.0 LITERATURE REVIEW

2.1 Use of Medicinal plants as Immunostimulants in Aquaculture

Although medicinal plants have been used as immunostimulants for thousands of years

(Tan andVanitha, 2004), the immunostimulating activity of herbal components has been

most widely studied in mice, chickens or human cell lines (Cao and Lin, 2003; Lin and

Zhang, 2004; Lin et al., 2006; Shan et al., 1999). In aquaculture, medicinal plants are also

used as chemotherapeutics and feed additives (Chang, 2000). They have the properties of

growth promoting ability, a tonic to improve the immune system, antimicrobial

capability, and stimulating appetite and anti-stress characteristics (Citarasu, 2010).

Several plants or their byproducts contain phenolic, polyphenolic, alkaloid, quinone,

terpenoid, lectine, and polypeptide compounds, many of which are effective alternatives

to antibiotics, chemicals, vaccines, and other synthetic compounds (Harikrishnan et al.,

2011a). In addition, medicinal plants are rich in a wide variety of nutrients (Chang,

2000). They can be administered as a whole plant or parts (leaf, root or seed)or extract

compounds, via water routine or feed additives, either singly or as a combination of

extract compounds, or even as a mixture with prebiotics or other immunostimulants. A

better understanding of modes of action may lead to effective and appropriate

applications of medicinal plants in aquaculture, because effects of administration of

herbal adjuvant may be species specific (Zakęś et al., 2008). Unfortunately, the mode of

action is not always addressed, especially in molecular mechanism levels.

7
2.2 Plants used as Immunostimulants in Aqauculture

Medicinal plants have been used as immunostimulants for thousands of years (Tan and

Vanitha, 2004) as mentioned above. The application of natural and innocuous compounds

has potential in aquaculture as an alternative to the use of antibiotics. A diverse range of

herbs is used in aquaculture (Table 1) including aloe (Aloe vera) (Kim et al., 1999),

almond (Terminalia catappa) (Chitmanat et al., 2005), basil flowers (Ocimum sanctum)

(Peraza-Gómez et al., 2009, 2011), caraway seed meal (Carum carvi L.) (Ahmad and

Abdel-Tawwab, 2011), cinnamon (Cinnamomum zeylanicum) (Ahmad et al., 2011),

garlic (Allium sativum) (Aly and Mohamed, 2010; Talpur and Ikhwanuddin, 2012),

ginger (Zingiber officinale) (Dügenci et al., 2003; Punitha et al., 2008; Talpur et al.,

2013), ginseng (Panax ginseng; Ginsana® G115) (Goda, 2008), American ginseng (P.

quinquefolium) (Abdel-Tawwab, 2012), green tea (Camellia sinensis L.) (Abdel-Tawwab

et al., 2010; Hwang et al., 2013), Arabic coffee bean, Coffee Arabica (Abdel-Tawwab,

2015a), guava (Psidium guajava) (Direkbusarakom, 2004), heartleaf moonseed

(Tinospora cordifolia) (Sudhakaran et al., 2006), olive tree leaf (Olea europaea) (Micol

et al., 2005), papaya leaf (Penaflorida, 1995), peppermint (Menthapiperita)

(Mousavietal.,2011), purple cone flower (Echinacea purpurea) (Aly and Mohamed,

2010), quillaja saponins (Quillaja saponaria) (Francis et al., 2005), seaweeds (Sargassum

spp.) (Immanuel et al., 2004, 2010), and tulsi (Ocimum sanctum) (Logambal et al., 2000).

According to Bulfon et al. (2015), more than 60 different medicinal-plant species have

been studied for the improvement of fish health and disease management in aquaculture.

Whole or parts of medicinal plants can be used for extracting medicinal compounds.

Whole plant Cynodon dactylon was used to prevent the white spot syndrome virus

8
(WSSV) infection in black tiger prawns (Balasubramanian et al., 2008), while

Rosmarinus officinalis was mixed with the feed either as whole dried leaves or as dried

ethyl acetate extract (Abutbul et al., 2004). Common parts can be used viz. fruit (Massa

medicate) (Takaoka et al., 2011),fruit (Piper guineese) and seed (Xylopia aethiopica)

(Okeke et al., 2001), leaves (Logambal et al., 2000; Micol et al., 2005; Sudhakaran et al.,

2006), roots (Wang et al., 2011), seeds (Kirubakaran et al., 2010; Sivaram et al., 2004),

flowers (PerazaGómezetal., 2011), or powders, which can be purchased from markets

(Kim et al., 1999; Radhakrishnan et al., 2014; Sahu et al., 2008). Some herbs used in

aquaculture are in the form of extract compounds such as Astragalus radix (from

Astragalus membranaceus) and Scutellaria radix (from Scutellaria baicalensis) (Yinetal.,

2006), and anthraquinone extract(from Rheum officinale Bail) (Liuetal., 2010) are

commonly used in aquaculture because of their ability to enhance fish immune systems.

The extracts vary and depend on herbal species, e.g. between 10.20% and17.50%

(Immanuel et al., 2004). Astragalus extract (containing 40% of Astragalus

polysaccharide) and Scutellaria extract (containing 20% of baicalin) were the commercial

products from Xuancheng Baicao Plants Industry and Trade Ltd. China (Yinetal., 2006).

The extract processes are simple (Sivaram et al., 2004). The extraction can be done with

acetone (Punitha et al., 2008), benzene (Balasubramanian et al., 2008; Punitha et al.,

2008), butanol (Praseetha, 2005; Punitha et al., 2008), ethanol (Adigüzel et al., 2005;

Harikrishnan et al., 2011c; Hwang et al., 2013; Sudhakaran et al., 2006; Wang et al.,

2011), ethyl acetate (Abutbul et al., 2004), methanol (Adigüzel et al., 2005;

Balasubramanian et al., 2007; Citarasu et al., 2003), hexane (Adigüzel et al., 2005),

petroleum ether (Balasubramanian et al., 2007; Punitha et al., 2008; Sudhakaran et al.,

9
2006), diethyl ether, chloroform, ethyl acetate (Balasubramanian et al., 2007), or even

boiling in hot water (Immanuel et al., 2010), boiling in water and then fermented by

yeasts (Takaoka et al., 2011). Various chemicals may lead to different degrees of effects

of medicinal plants on aquaculture. Alcoholic or organic solvents provide a higher

efficiency in extracting secondary bioactive metabolites (polar or non-polar) with

antimicrobial and immunostimulant activity, compared to water-based methods (Bulfon

et al., 2015).

Among ethanol, methanol and hexane extracts from Ocimum basilicum, the hexane

extract showed a stronger and broader spectrum of antibacterial activity against 146

microbial organisms including aquaculture pathogens (Adigüzel et al., 2005). The hexane

soluble fraction of Solanum trilobatum was more protective than the water soluble

fraction when administered intraperitoneally to tilapia (Divyagnaneswari et al., 2007).

Clitoria ternatea extracted using ethyl acetate, ethanol, acetone and petroleum ether

showed higher antibacterial effects against a range of fish pathogens than that extracted

using water (Ponnusamy et al., 2010). Administration of triherbal aqueous, ethanol or

methanol solvent leaf extracts enhanced immune parameters and disease resistance

against Aeromonas hydrophila in goldfish (Carassius auratus), but the ethanol solvent

extract appeared more effective as an immunostimulant (Harikrishnan et al., 2009a). In

the same plants, ethanol extracts had a greater inhibitory activity in vitro against

Streptococus agalactiae than the aqueous extracts did (Rattanachaikunsopon and

Phumkhachorn, 2009).

10
 Table 1: Herbal plant families & their multiple therapeutic properties in

aquaculture

Scientific name Family Therapeutic Properties Source

Myristica fragrans Myristicaceae Digestion, Stimulant & Coutteau et al.,
Antidiarrhoic 2011
Eclipta alba Asteraceae Immunostimulatory effect Pandy Govind et
al., 2012
Datura metal Solanaceae Antimicrobial effect Ravikumar et al.,
2010
Allium sativum Amaryllidaceae Immunostimulant Nargis et al., 2011
Nymphaea alba Nymphaeaceae Antibacterial Turker et al., 2009
Zingiber officinale Zingiberaceae Phagocytosis of WBC Yin et al., 2008
Vitex negundo Lamiaceae Antibacterial agent Prabhu Narayan
Marimuthu et al.,
2012
Solanum nigrum Solanaceae Anticarcinogenic Patel et al., 2009
Azadirachta indica Meliaceae Antiviral, antiseptic, Cristea et al., 2012
fungicidal
Curcuma longa Zingiberaceae Anti-inflammatory, Chattopadhyay et
antifungal, anti-venom, al., 2004
Anticoagulant, Antifertility

Vernonia amygdalina Asteraceae Antibacterial, Huffman et al, 1989

immunostimulants

2.3 Administration and Application Methods of Immunostimulants

Similar to other immunostimulants, medicinal plants can be applied via injection

(Harikrishnan et al., 2011a), bathing/immersion (Çek et al., 2007) or oral administration

(Harikrishnan et al., 2011a). The latter seems to be the most practical (Jeney and

Anderson, 1993; Sakai, 1999; Yin et al., 2006). Administration of herbs can be achieved

singly or in combination. Both methods have the same degree of use and practicality.

Herbal extracts have been commonly used as feed additives in aquaculture (Wang et al.,

2015). As with other immunostimulants, a combination of herbs provides beneficial

11
effects to hosts. Astragalus and Lonicera extracts alone or in combination enhanced

respiratory burst and phagocytic activity of blood phagocytes and plasma lysozyme

activity (Ardó et al., 2008).

A mixture of herbs Angelica membranaceus and A. sinensis increased respiratory burst

activity of phagocytic cells and plasmalysozyme activity of common carp (Cyprinus

carpio) (Jian and Wu, 2004) and large yellow croaker (Pseudosciaena crocea) (Jian and

Wu, 2003), rainbow trout (Oncorhynchus mykiss), Indian catla carp (Catla catla) (Dey

and Chandra, 1995) and Mozambican tilapia (Oreochromis mossambicus) (Logambal and

Michael, 2000). Purple coneflower and garlic supplemented diets acted as

immunostimulants for Nile tilapia (Oreochromis niloticus) (Aly and Mohamed, 2010).

Moreover, a practical application can be a combination of more than two herbal species

or with commercially manufactured herbal products. The application of the traditional

Chinese medicine formulation of four herbs at the ratio of 1:1:2:3 would be a

prophylactic approach for disease control, replacing the use of antibiotics for treating

enteritis and even other general diseases in grass carp (Ctenopharyngodon idellus) culture

(Choietal., 2014). The herbal mixture of Massamedicata fermentata, Crataegi fructus,

Artemisia capillaries, and Cnidium officinale at the proportion of 2∶2∶1∶1 enhanced the

weight gain and feed efficiency of Japanese flounder (Paralichthys olivaceus) (Seung-

Cheol et al., 2007). A combination of essential oils from the herbs (Thymus vulgaris,

Salvia officinalis, Eucalyptus globules and Mentha piperita) had potent antibacterial

effects against a range of bacterial species (Staphylococcus aureus, Escherichia coli and

Pseudomonas aeroginosa) (Mousavi et al., 2011). A diet mixture of equal proportions of

five herbs (Acalypha indica, Hygrophila spinosa, Picrorhiza kurooa, Tinospora

12
cordifolia and Zingiber officinale) improved the total hemocyte count, phagocytosis,

phenol oxidase activity, haemagglutinin activity and bacterial clearance of Indian white

prawns, Fenneropenaeus indicus (Rajeswari et al., 2012). A mixture of equal proportions

of six herbs and plant materials in diets enhanced or impaired enzyme activity (Lin et al.,

2006).

A diet of rotifer enriched with a herbal mixture of Massa medicata, Crataegi fructus,

Artemisia capillaries, and Cnidium officinale promoted growth and resistance against

Vibrio anguillarum in red sea bream (Pagrus major) larvae (Takaoka et al., 2011). The

plant mixture basil flowers and commercial antiviral plants (VPH®, HSV®, Amazonas

Herbs Distrito Federal, Mexico) provided protection for white leg prawns (Litopenaeus

vannamei) against WSSV (Peraza-Gómez et al., 2009, 2011). A combination of herbs

and micronutrients like selenium, boron or zinc has provided several benefits to hosts. A

combination of Chinese herbs either Astragalus membranaceus or Lonicera japonica

with boron enhanced the immune response of Nile tilapia and resisted against Aeromonas

hydrophila (Ardó et al., 2008). Bacteriolytic activity and leukocyte function were

improved by mixtures of chosen Chinese herbs incorporated in fish diet (Chansue et al.,

2000).The best survival rate was observed in Nile tilapia treated with both herbs and

boron (Ardó et al., 2008). In addition, another synbiotic can be seen between herbs and

probiotics.

The combination of medicinal herb and Bacillus bacteria provided a synergistic effect on

the growth rate of white leg prawns, because Bacillus bacteria accelerated the prawn

ability to absorb the medicinal herb, while the metabolites from medicinal herb could be

used by Bacillus bacteria (Yuet al., 2008, 2009). The methods of treating microbial

13
 diseases in fish are problematic, neither effective nor cost efficient, because a large

amount of chemotherapeuticagentsisneededandthendischargedintotheenvironment, which

poses a risk to animals and human health.

2.4 Mode of Action of Immunostimulants

2.4.1 Improve the Innate Immune Responses

Several herbal medicines showed an anti-microbial activity, and facilitated growth and

maturation of various fish species (Harikrishnan et al., 2011a). Therefore, there is a

growing interest in using medicinal herbs as immunostimulants in aquaculture. Many

investigations on their effects at molecular mechanism levels have been undertaken, for

instance on Astragalus membranaceus and Nelumbo nucifera (Liu et al., 2004; Shao et

al., 2004). The use of immunostimulants is of an increasing interest for boosting the

defense mechanisms and conferring protection of animals from infectious diseases.

Immunostimulants enhance the innate (or non-specific) immune response (Sakai, 1999).

The major components of the innate immune system are macrophages, monocytes,

granulocytes and humoral elements, like lysozyme (Magnadóttir, 2006). Herbs contain

many types of active components, like polysaccharides, alkaloids or flavonoids (Ardó et

al., 2008). The herbal-compound extracts act as immunostimulants to enhance the

immune response of fish (Hardie et al., 1991; Siwicki, 1989; Thompson et al., 1993) viz.

lysozyme, complement, antiprotease, meloperoxidase, reactive oxygen species, reactive

nitrogen species, phagocytosis, respiratory burst activity, nitric oxide, total hemocytes,

glutathione peroxidase, and phenoloxidase, against bacterial, fungal, viral, and parasitic

diseases(Harikrishnanetal.,2011a).

14
 The use of Prunella vulgaris extract enriched diets for olive flounder, the scuticocidal

activity and respiratory burst activities increased (Harikrishnan et al., 2011b).The main

active component of Astragalus extract is a polysaccharide (Ardó et al., 2008). This

Astragalus polysaccharide is a well-studied immunostimulant (Tan and Vanitha, 2004).

As immunostimulants enhanced the plasma lysozyme activity (Hanif et al., 2005; Kim

and Austin, 2006), elevated lysozyme level was increased when large yellow croaker

(Jian and Wu, 2003) and common carp (Jian and Wu, 2004) were fed various Chinese

herbal extracts. The addition of green tea ethanol extract to the diet improved lipid

utilization, lysozyme activity and stress recovery, and reduced total cholesterol levels of

black rockfish (Hwang et al., 2013). Phagocytic cells are the most important cellular

components of the innate immune system of fish (MacArthur and Fletcher, 1985).

Phagocytes also produce toxic oxygen forms during a process called respiratory burst

(Neumann et al., 2001). Phagocytic activity is a primitive defense mechanism (Neumann

et al., 2001) and an important characteristic of the fish immune system (Seeley et al.,

1990). This parameter usually shows an increase after oral administration of

immunostimulants (Jeney et al., 1997; Siwicki et al., 1994). Herbal medicine extracts can

enhance phagocytosis in various fish species (Dügenci et al., 2003).

2.4.2. Enhance Antimicrobial Activity

The antimicrobial properties of medicinal plants and their active compounds have been

investigated by many researchers world-wide. Herb extracts have great anti-bacterial

activity against both Gram positive and Gram negative bacteria. They can even be used to

treat specific diseases caused by virus, parasites and fungi. Several compounds extracted

15
 from medicinal plants inhibited the growth of Gram-positive and Gram-negative

organisms (Harikrishnan et al., 2009b).

2000). Being immunostimulants, herbs act as anti-viral agents to the host immune

system. It is assumed that the herbal active compounds inhibited or blocked the

transcription of the virus to reduce the replication in the host cells, and enhance non-

specific immunity (Citarasu, 2010). Herbs have been found to have antiviral properties

against fish viruses (Direkbusarakom et al., 1996c).

2.4.3. Herbal Plants as Promising Antibiotics

Several studies have proved that herbal plants can be used as promising antibiotics that

after challenging with pathogens, the survival rates of infected fish priority fed various

immunostimulants (Sakai, 1999), vaccines (Gudmundsdóttir and Magnadóttir, 1997) and

probiotics (Brunt et al., 2007), increased. After challenge with A. hydrophila, the best

survival rate was observed in fish treated with herb and boron (Ardó et al., 2008). The

methonolic extracts of three ayurvedic herbals viz. Solanum trilobatum, Andrographis

paniculata, and Psoralea corylifolia showed the protection of Penaeus sp. against nine

pathogens such as Bacillus subtilis, Proteus vulgaris, Salmonella typhi, K. pneumoniae,

Pseudomonas aeruginosa, P. fluorescens, Vibriosp., S. aureus and A. hydrophila

(Citarasu,2000).

Butanolic extract of Withania somnifera through Artemia enriched diet successfully

controlled Vibrio parahaemolyticus and V. damsela infection in prawns (Praseetha,

2005). Psidium guajava could control disease caused by A. hydrophila in Nile tilapia

(Rattanachaikunsopon and Phumkhachorn, 2009). Green tea, cinnamon, and American

16
 ginseng improved the resistance of Nile tilapia against A. hydrophila infection (Abdel-

Tawwab, 2012; Abdel-Tawwab et al., 2010; Ahmad et al., 2011).

2.4.4. Enhance the fish growth, Feed Utilization and Nutrient Digestibility

Medicinal plants have been proven as growth promoters. Firstly, they enhance digestive

enzymes, and thus boost survival and growth rates of aquatic animals. Three herbs

(Alteranthera sessilis, Eclipta alba and Cissus quadrangularis) acted as appetizers and

enhanced the activities of digestive enzymes (protease, amylase and lipase) of freshwater

prawns (Radhakrishnan et al., 2014). This resulted in an enhancement of food utilization

and ultimately in the production of better growth rates as indicated by the evidence of

elevated concentrations of vitamins, protein, essential amino acids, unsaturated fatty

acids and minerals. Vitamin C and Elevels in the hepatopancreas, and both sodium and

potassium levels, and muscle of freshwater prawns increased when the prawns were fed

diets supplemented with herbs (Radhakrishnan et al., 2014). American ginseng, green tea

and cinnamon enhanced the growth performance and feed utilization of Nile tilapia

(Abdel-Tawwab, 2012; Abdel-Tawwab et al., 2010; Ahmad et al., 2011). Caraway seed

meal improved the growth performance and feed utilization of Nile tilapia (Ahmad and

Abdel-Tawwab, 2011). Caffeine improved the growth of sea bream (Sparus aurata) at a

concentration higher than 0.1% diet (Chatzifotis et al., 2008). The herbal mixture of

Massa medicata fermentata, Crataegi fructus, Artemisia capillaries, and Cnidium

officinale enhanced the growth, and fatty acid utilization of Japanese flounder (Seung-

Cheol et al., 2007).

Papaya leaf meal contains an enzyme, papain, which increased protein digestion, FCR,

SGR, and weight gain of black tiger prawn PL (Pena florida, 1995). A caffeine diet at

17
 0.2–0.5% increased the FCR of sea bream (Chatzifotis et al., 2008). The FCR of

Macrobrachium rosenbergii fed with anthraquinone extracts was lower compared to

those without anthraquinone extracts (Liu et al., 2010). In addition, the administration of

Tribulus terrestris extract led to an increase in sex reversal ratio because it stimulated

permatogenesis, and exhibited as a growth accelerator (Çek et al., 2007).

2.5 Neem-Azadirachta indica

A. indica is a member of the family Meliaceae (Kashif and Ullah, 2013) and is native to

the Indian sub-continent (Gajalakshmi, 2002) and has been used extensively in Asian and

African subcontinent because of its medicinal properties (Srivastava and Prakash, 2006).

Almost every part of A.I. tree has been known to possess a wide range of

pharmacological properties (Farah et al., 2006; Van Wyk and Wink, 2004). In recent

years A.I. has attracted global attention due to its potential as a source of natural drugs

(Kumar, 2002; Gajalakshmi and Abbasi, 2004) because of the presence of triterpenoids,

steroids, carotenoids, ketones and phenolic compounds (Jacobson, 1990),

Azadirachtin A, B, D, H, I, Desacetylnimbin, Azadiradione, Nimbin, Salanin, Azadirone,

Nimbolin, Nimbinene, Nimbolide (Harikrishnan et al., 2003; Sadeghian and

Mortazaienezhad, 2007). Neem Gum is a rich source of protein (Kashif and Ullah, 2013).

2.5.1 Azadirachta indica usage in aquaculture

In the field of aquaculture A. indica has been reported to enhanced primary and

secondary antibody response in Oreochromis mossambicus (Logambal and Michael

2000, 2001); to control fish predators in macroinvertebrates (Dunkel and Ricilards,

1998); as alternative for the control of parasites and predators such as dragon-fly larvae in

18
 Prochilodus lineatus (Martinez, 2002); to assess acute lethal and sublethal effects on

Prochilodus lineatus (Winkaler et al., 2007); to possess antibacterial effect in Channa

striatus (Abdul Kader Mydeen and Haniffa, 2011); to exhibit antibacterial activity in

ornamental fishes (Ravikumar et al. 2011); to exhibit antiviral properties in cultured

shrimp (Banerjee et al., 2013); to produce disease resistant fry of Catla catla (Rao et al.,

2004); for assessment of 72-hr Median Lethal concentration in Cyprinus carpio Juvenile

(Davoodi, 2012); for assessment of growth in Tilapia zilli, when exposed to sublethal

concentrations (Omoregie and Okpanachi, 1992); for assessment of acute toxicity in

Tilapia zilli (Omoregie and Okpanachi, 1997); as an effective inhibitory agent of

reproduction in Tilapia zilli (Jegedel and Fagbenro, 2008).

2.6 Bitter Leaf –Vernonia amygdalina

Vernonia is the largest genus of the tribe Vernonieae with close to 1000 species; it occurs

mainly in South America and Africa ( Burkill, 1985). More than 300 species have been

described from Africa with about one third occurring in Madagascar. Apart

from Vernonia amygdalina several species are eaten as vegetable, of which Vernonia

hymenolepis is most important. Vernonia colorata (Willd.) W.F.M.Drake is closely

related to Vernonia amygdalina. It differs in its more or less entire leaves and glabrous

fruits. Leaves of Vernonia colorata are mainly collected from the wild, and its primary

use is as a medicinal plant. Other species occasionally cultivated as a vegetable but more

often collected from the wild are Vernonia cinerea (L.) Less (Misari, 1992). in

Kenya, Vernonia poskeana Vatke & Hildebrandt in Zimbabwe, which are both also more

important as medicinal plants, Vernonia appendiculata Less. in Madagascar

and Vernonia perrottetii Walp. in Sierra Leone (Beentje, 2000).

19
 CHAPTER THREE

3.0 MATERIALS AND METHODS

3.1 Location of the Study

The experiment was conducted at the fisheries unit of the Experimental farm of the

Department of Fisheries Technology, College of Agriculture, Science and Technology,

Lafia. Lafia is located on Latitude 8 0 35’N, longitude 80 32’E, altitude 181.53m above sea

level with a mean temperature of 26.70 C, relative humidity of 75-87% and average day

light of 9-12h (NIMET, 2021).

3.2 Procurement of Experimental Fish

Clarias gariepinus day old fry were purchased from a reputable fish farm in Lafia. The

specimen was weighed and acclimatized in a well aerated de-chlorinated water for a

period of ten days.

3.3 Collection and processing of plant extracts bitter leaf (vernonia amygdalina) and

neem leaves (Azadirachta indica)

Bitter leaf and neem leaves were purchased at the New market in Lafia L.G.A of

Nasarawa State, Nigeria. The leaves was plucked, chopped separately into smaller sizes

and subsequently was dried under shade at room temperature until the leaves was fully

dried. The dried leaves was pounded separately using mortar and pestle into fine powder

and was then sieved using siever to obtain fine powder The powdered leaves was stored

separately in an air tight bottle until required. Methanol extraction of the leaves was done

using soxhlet apparatus.

20
3.3 Experimental Set Up

The experiment consisted of three treatments replicated three times in a completely

randomized design. Ninety (90) Clarias gariepinus day old fry was used for the study.

The fish were stocked at 10 fingerlings per bowl. The treatments were labeled thus T1

(control), T2, and T3 respectively.

3.4 Data Collection

Fish performance were determined using the following formulae:

3.4.1 Weight gain

This is calculated using the formula
Wf – Wi
Where, Wf is final mean weight and Wi is initial mean weight
3.4.2 Percentage mean weight gain (PMWG)
This is calculated using the formula

PMWG = (Wf - Wi) x 100

Wi
Where, Wf is final mean weight and Wi is initial mean weight
3.4.3 Specific growth rate (SGR)
This is the percentage rate of change in the logarithmic body weight. The SGR was

calculated using the formula below:

SGR= (1n Wf – 1n Wi ) x 100

T (days)

Where, Wf is final mean weight, Wi is initial mean weight and T is culture period.

21
3.4.4 Length

This was taken by measuring the total length from the tip of the snout to the end of the

caudal fin using a ruler calibrated in centimeters.

3.4.5 Survival rate

Survival rate % = No of fish survived at the end of experiment x 100
Initial number of fish stocked
3.4.5 Physico-chemical parameters

Temperature of water was measured daily using mercury in glass thermometer, pH of

water was measured daily using a pH meter while dissolved oxygen was determined

using the methods of AOAC (2000).

3.5 Statistical Analysis

Data collected in this experiment was analysed using Analyses of variance (ANOVA)

and significant mean was separated at 0.05% probability level as described by (Steel et

al., 1997).

22
 CHAPTER FOUR

4.0 RESULTS AND DISCUSSION

4.1 Results

The result of the study conducted on the Clarias gariepinus fry showed that the highest

weight gain was recorded in Bitter Leaf (200ml/l) and the lowest in control diet. The

highest mortality was recorded in control diet and the lowest in the diet 4 (Bitter leaf +

Neem leaf). The highest specific growth rate was recorded in Bitter leaf and the lowest in

control diet. The highest level of Survival was recorded in Bitter leaf (200ml/l) while the

lowest was recorded in the control.

23
 Table 1: Growth performance of Clarias gariepinus fry Treated with bitter leaf and Neem leaf extract.

Control Bitter leaf extract Neem leaves extract Bitter Leaf/ Neem leaves

Trt 1 Trt 2 (150ml/l) Trt 3 Trt 1 Trt 2 Trt 3 Trt 1 Trt 2

(100ml/l) (200ml/l) (100ml) (150ml) (200ml) (150ml) (200ml)
Initial 4.50 + 0.00a 4.50 + 0.01a 4.50 + 0.01a 4.50 + 0.01a 4.50 + 0.00a 4.50 + 0.00 a
4.50 + 0.00a 4.50 + 0.00a 4.50 + 0.00a
Body (g)
Final 10.23 ± 0.01b 12.21 ± 0.02a 12.57± 0.01a 12.62 ± 0.00a 11.92± 0.01a 12.21±0.02a 12.40±0.01a 12.33±0.01a 12.30±0.01a
Body (g)
Weight 5.73 ± 0.01b 7.7 ± 0.03a 8.07 ± 0.02a 8.5± 0.01a 7.42 ± 0.03a 7.71 ± 0.03a 7.9 ± 0.05a 7.83 ± 0.02a 7.80 ± 0.03a
gain (g)
Weight 127.33±0.35b 171.11±0.54a 179.33±0.66a 186 ± 0.78a 164.89±0.97a 171.33±0.67a 175.56±0.58a 174.00±0.35a 173.33±0.45a
gain (%)
Specific 1.48±0.04b 1.79±0.07a 1.84±0.06a 1.89 ± 0.05a 1.75±0.07a 1.79±0.07a 1.82±0.06a 1.80±0.06a 1.80±0.06a
growth
rate
The above values are means of duplicate data, mean values in each row with similar superscripts are not significantly different.

24
Table 2: Disease Resistance of Clarias gariepinus fry treated with bitter leaf and neem leaf extract

Control Bitter leaf extract Neem leaves extract Bitter Leaf/ Neem leaves

Trt 1 Trt 2 Trt 3 Trt 1 Trt 2 Trt 3 Trt 1 Trt 2

(100ml/l) (150ml/l) (200ml/l) (100ml/l) (150ml/l) (200ml) (150ml) (200m)
Survival 89.00 98.00 98.33 98.66 97.00 97.65 98.00 98.00 97.45
(%)
The above values are means of duplicate data, mean values in each row with similar superscripts are not significantly different.

25
Water quality parameters of the sources of water in this study

The obtained result of pH values, dissolved oxygen (mg/L) total alkalinity (mg/L)

temperature (0C) and free carbon dioxide(mg/L) of each of the treatment on this research

work are presented in Table3: The result shows that all the water quality parameters from

sources of water supplied for the treatment were not significantly different (P>0.05) from

each other.

26
Table 3: Water quality parameters of Clarias gariepinus fry exposed to neem leave and
Bitter leaf extract

Parameters Control NLE BLE NLBLE

Temperature (°C) 27.20+0.04 27.23+0.06 27.32+0.02 27.31+0.05

pH 7.25+0.02 7.24+0.0.03 7.25+0.0.02 7.24+0.02

Total alkalinity 15.22+0.01 15.22+0.01 15.22+0.03 15.22+0.02

DO (mg-l) 7.22+0.03 7.21+0.0.03 7.22+0.03 7.22+0.03

CO2 (mg-l) 4.11+0.01 4.10+0.03 4.11+0.02 4.11+0.01

NLBLE = Neem leave and Bitter extract

NLE = Neem leave extract

BLE = Bitter leaf extract

27
4.2 Discussion

The growth and survival of Clarias gariepinus fry is largely dependent on good

treatment and disease resistance of the fish. the use of Natural plants extracts to serve as

prophylactic treatments of the fish can serve a better source of prophylaxis than use of

foreign medications. The most important secondary substances in medicinal plants

include, alkaloids, glucosides, steroids, flavonoids, fatty oils, resins, mucilages, tannins,

gums, phosphorus and calcium Kubmarawa et al., [2008]. Medicinal values of secondary

metabolites are due to the presence of these chemical substances that produce a definite

physiological action on target organism. The leaves, barks, fruits and roots of the Neem

plant have been highly appraised for their medicinal purposes.

The tannins, alkaloid, steroids, saponin and flavonoids have been identified in fresh and

dried leaves of the Neem and Bitter leaf plant Trease and Evans, [1989]; Susmitha et al.,

[2013]. Positive reaction and subsequent quantitative content of these metabolites in the

studied leaf extract therefore followed the previous trend, in which these compounds

were qualitatively and quantitatively detected.

The proximate composition of the experimental diets used in this study supported the

growth of C. gariepinus juvenile as reported by Olaifa, Ajayi, Taiwo and Bello (2012);

Olaifa and Bello (2011) and Eyo (1995) that for maximum growth, fry, fingerlings and

juveniles must have a diet in which nearly half of the digestible ingredients consist of

balanced proteins. The first mortality was recorded in the control experiment on the 7th

day. The observed symptoms of this infection included lack of appetite, swimming

abnormalities, pale gills, bloated appearance, and skin ulcerations. These syndromes were

similar to those reported by Zhang, Gong, Yu and Yuan (2009).

28
The results shows that all the water quality parameters from the sources of water supplied

for the treatments were within the acceptable range according to WHO, (2010). The

temperature recorded in the present study were (27.21 + 0.04, 27.23+ 0.06, 27.31+0.02,

and 27.32+ 0.05)are in the optimum growth and survival rate of Clarias gariepinus fry

according to Boyd (1990). It has been observed the fish grows best at temperature

between 250C and 27oC by Boyd and Lichtkopper (1979) and Dupree and Itunner (1984).

The pH range of 7.25+0.02, 7.24+0.03, 7.24+0.02, and 7.25+0.02 recorded in the study

with the recommended range of 0.5+0.09 (Boyd and Lichtkopper, 1997).

This result is in range with the result presented by Uzuku et al, 2015. Which is between

7.30+0.03 and 7.57+ 0.03 Stirling and Philip (1990) gave a pH range of 5.0-9.5 as

suitable pH for aquatic lives for fishes.

The pH values of (NLBLE) neem leave and bitter leaf extract of the each different

treatment were not significantly different from each other because water suppllied for

each treatments were the same and partly because pH is a conservative parameters in

buffered media (Ubong and Gobo, 2001).

The dissolved oxygen 7.21+0.03, 7.21+0.03, 7.22+0.03, and 7.22+0.03 recorded in the

study are consider between the recommended range of fish production. This view was in

agreement with (Masser et al., 1999) who reported that the permissible range of dissolved

oxygen concentration for warm water fish culture 5.0 to9.5mg/L.

The carbon dioxide (mg/l recorded in this experiments range from 4.10 + 0.01,

4.10+0.03, 4.11+0.02 and 4.11+0.01 are significantly not different from each other.

Swann (1997) suggested that fish can be tolerated under the concentration of C0 2 of about

29
10ppm provided dissolve oxygen (DO) concentration are high. According to Ekubo and

Abowei (2011). Therefore if tropical fish can tolerate CO 2 over 100mg/l. this dearly

shows that Clarias gariepinus fry are within the recommended range of survival.

The mean value of total alkalinity (mg/l) ranges between 15.22+0.01, 15.22+0.01,

15.22+0.02 and 15.22+0.03 is not (P>0.05)significantly different from each treatment of

(NLBLE) neem and bitter leaf extract. These result were also in agreement with those

represented by Okunsebor et al.,(2015) which ranges between 15.22+0.01, 15.22+0.01,

15.22+0.02, 15.22+0.03 and 15.22+0.02 according to Murphy(2008). Total alkalinity

level of 20-200ppm are typical fresh water and will stabilize the pH level in a pond level

below 10ppm.

30
 CHAPTER FIVE

5.0 SUMMARY, CONCLUSION AND RECOMMENDATIONS

5.1 Summary

Disease is considered as one of the important problematic factors for the seed industry. It

was found that parasitic diseases are particularly one of the most important limiting

factors for growth and survival of fish fry and fingerling. The management and control of

parasitic and other infections in aquaculture are a constant challenge. The analysed Neem

leaf and Bitter leaf has secondary metabolites associated with antimicrobial properties in

varied quantities. These chemical metabolites were capable of mildly curtailing mortality

in yolk-fry stage of C. gariepinus.

5.1 Conclusion

From the result of this study, it is therefore concluded that the prophylactic property was

recorded in the Bitter leaf Treatment. The study therefore reveals bitter leaf as a better

compared to the control and Neem leaf extracts. The use of bitter leaf and neem leaves

extract as a prophylactic treatment for C.gariepinus fingerlings will serve as an effective

method of diseases resistance.

5.3 Recommendation

The following recommendations were made:

1. Research should be intensified to exploit more natural plants to be used for

prophylaxis of fish.

31
2. Fish farmers are advised to use natural plants extracts for better C. gariepinus fry

culture.

3. Mixture of plant extracts to enhance survival and growth of fish hatchlings should be

highly encouraged.

32
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