EFFECT OF MANGIFERA INDICA L. (MANGO) KERNEL ON CLARIAS GARIEPINUS (AFRICAN CATFISH) FINGERLINGS INFECTED WITH AEROMONAS CAVIAE


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TABLE OF CONTENTS

TITLE PAGE
ABSTRACT
TABLE OF CONTENTS
ABBREVIATION

CHAPTER 1: INTRODUCTION
1.1       Background of the Study
1.1.1    Aeromonas caviae
1.1.2    Mango (Mangifera indica)
1.2       Statement of Research Problem
1.3       Justification
1.4       Aim of the Study
1.5       Specific Objectives of the Study were:
1.6       Research Hypotheses

CHAPTER 2: LITERATURE REVIEW
2.1       Fish Production
2.1.1    Biology and history of African catfish
2.1.2    Taxonomy
2.1.3    Nature and geographical distribution
2.1.4    Habitat
2.1.5    Natural food and feeding
2.1.6    Natural reproduction
2.2       Mangifera indica L. (Mango)
2.2.1    Origin and distribution
2.2.2    Taxonomy and variety
2.2.3    Size
2.2.4    Canopy
2.2.5    Roots
2.2.6    Flowers
2.2.7    Leaves
2.2.8    Fruit
2.2.9    Seed
2.2.10 Pharmacology of M. indica seed (kernel)
2.2.11 Phytochemistry of M. indica kernel
2.3       Aeromonads
2.3.1    Taxonomy
2.3.2    Historical aspects of aeromonads
2.3.3    Isolation of aeromonas
2.3.4 Biochemical appearance and identification
2.3.5 Aeromonas and ecosystem
2.3.6 Diseases caused by aeromonas in fish
2.3.7 Pathology
2.3.8 Virulence factors
2.3.9 Serology and vaccination
2.3.10 Epidemiology
2.3.11 Zoonoses
2.3.12 Methods of control
2.3.13 Antimicrobial susceptibilty

CHAPTER 3: MATERIALS AND METHODS
3.1       Identification of Mango Kernel
3.2       Extraction
3.2.1    Preparation of the aqueous extract of mango kernel
3.2.2    Preparation of methanol extract of mango kernel
3.2.3    Phytochemical screening
3.2.4    Test for carbohydrates:
3.2.5    Test for anthraquinones:
3.2.5.1 Test for free anthracene derivatives (Bontrager’s test)
3.2.6 Test for unsaturated steroid and triterpene:
3.2.7 Test for cardiac glycoside:
3.2.7.1 Keller- Kiliani Test
3.2.8 Test for saponins:
3.2.9    Test for tannins:
3.2.10 Test for flavonoids:
3.2.11 Test for alkaloids:
3.3       Acute Toxicity Test
3.3.1    Limit test
3.3.2    Water quality test
3.4       Feeding Diet and Grouping of Fish
3.5       Preparation of the Herbal Diet
3.6       Collection of Blood and Serum
3.7       Antimicrobial Activity Test
3.8       Prophylactic Trials
3.9       Isolation of A. caviae from Infected Fish
3.9.1 Microscopic identification of A. caviae
3.9.2 Biochemical identification of A. caviae
3.10 Determination of Blood Parameters
3.11 Data Analysis

CHAPTER 4: RESULT
4.1 Phytochemical Constituent of M. indica kernel
4.2 Median Lethal Dose (LD50) of M. indica kernel Powder
4.2.1 Limit Test
4.3 Serum Antimicrobial Activity
4.4 Antimicrobial Activity Test of M. indica kernel Methanol Extract, Aqueous Extract and Standard Antimicrobials
4.5 Prophylactic Trials
4.6 Isolation of A. caviae from Infected Fish
4.7 Hematological Parameters

CHAPTER 5: DISCUSSION

CHAPTER 6: CONCLUSION AND RECOMMENDATION
6.1 Conclusion
6.2 Recommendations
APPENDICES




ABSTRACT


A number of approaches were employed to control diseases in fish with particular emphasis on use of chemotherapeutic agents. However, application of antibiotics in aquaculture is expensive and leads to antibiotics residue in fish, which could cause the development of antibiotic resistance in human. In this study, phytochemical analysis was done to detect the presence of secondary metabolites in the M. indica kernel. The in vitro and in vivo antimicrobial effects of aqueous and methanol extracts of mango kernel on A. caviae using agar well diffusion method were evaluated. In the in vivo trial, two hundred and fourty fingerlings were randomly divided into four groups of sixty. Fish in group A, B, C and D were fed diet containing 0, 1, 5 and 10 g/kg of mango kernel dry fish diet, respectively for 60 days. Some haematological parameters of fish were examined at 20 and 40 days of feeding the diet. Twelve fish from each group were challenged with A. caviae 60 days post feeding and clinical signs, mortalities and gross lesions were observed and recorded over 14 days post-infection. Phytochemical analysis of mango kernel revealed the presence of triterpenes, tannins, glycosides, saponins and flavonoids. The result obtained following the limit dose test demonstrates that the lethal dose (LD 50) of M. indica kernel powder is greater than 60 g/kg of feed. The methanol extract when used at concentrations that ranges from 50 to 500 mg/ml inhibited growth of the bacteria, with zone of inhibition ranging between 16 ± 2.41 to 24 ± 0.58. The antibacterial activity of the methanol extract was not significantly different from the standard antibacterial agents used in the study (gentamicin, enrofloxacin, neomycin, chloramphenicol and erythromycin). However, M. indica powdered kernel solution did show antibacterial activity against A. caviae at lower concentrations (166, 125, 100, 55.6 and 50 mg/ml) with smaller zones of inhibition that ranges from 8 ± 1.00 to 11 ± 1.00, with no measurable zones of inhibition at higher concentrations of 250 and 500 mg/ml. The results demonstrated that the serum of fish treated with mango kernel at different concentrations produced no antibacterial effect against A. caviae. However, the total leukocyte counts were significantly higher in fish treated with mango kernel at 10 g/kg of feed 40 days post-treatment. Less survivability was observed in fish that were not treated with diet containing mango kernel (50% survivability) up to day 14 after infection. The groups fed 5 and 10 g/kg mango kernel dry diet showed highest percentage survival (100%). Results of the present study clearly demonstrated that only methanol extract of mango kernel showed good antibacterial activity. The present study also confirmed the efficiency of the organic solvent for extraction of plant constituent compared to water. The study indicates that mango kernel protects Clarias gariepinus against A. caviae infection by enhancing the survivability of the treated fish.





CHAPTER ONE

INTRODUCTION

1.1 Background of the Study

Aquaculture  is  one  of  the  fastest     growing  food-producing  sectors  around  the  world

(Harikrishnan  et
at.,  2011). The world‟s total production of fish and shellfish (including
mollusk
and
crustacea) was 99 metric tonne in 1990 and it increased to 122  metric




tonne in 1997 (Hill, 2010). According to Food and Agriculture Organization (FAO), the global aquaculture production has increased from about 28.3 million tonnes to 40 metric tonnes in 2009 (FAO, 2009). According to Harikrishnan et al. (2011), among various kinds of cultivated organisms, many marine and freshwater finfish and shellfish species constitute an important industry with their production increasing every year. Aquaculture fish production increased significantly over the past few decades necessitating intensive fish culture practices.


In aquaculture, disease control using chemotherapeutic agents has been complicated by the misleading advice provided to the farmers by feed and chemical companies regarding the use of antibiotics and other therapeutic agents (FAO, 2003a). In the intensive aquaculture system, application of antibiotics and chemotherapeutics as prophylactic measures has been widely criticized for their negative impacts like immunosuppression, residue accumulation in tissues (Rijkers et al., 1980; FAO, 2003b; Harikrishnan et al., 2009a, 2009b) and the development of drug resistant pathogens and environmental pollution (Smith et al., 1994). International agencies recommends that the use of antibiotics should be restricted to therapeutic purposes only, and that preventive approaches should be preferred in fish disease management over costly post disease treatments (GESAMP, 1997; FAO, 2005).......


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