fisheries & aquaculture university of veterinary and animal sciences lahore

180 Pages·2015·1.12 MB·English

POSSIBLE CAUSES OF SELECTIVE LERNAEA ATTACK ON
DIFFERENT FISH SPECIES

FARZANA ABBAS

2007-VA-532

A THESIS SUBMITTED IN THE PARTIAL FULFILLMENT OF THE
REQUIREMENTS FOR THE DEGREE OF

OF
DOCTOR OF PHILOSOPHY
IN
FISHERIES & AQUACULTURE
UNIVERSITY OF VETERINARY AND ANIMAL SCIENCES
LAHORE- PAKISTAN
2014

i
DEDICATED

To

MY PARENTS

W
hose hands are always raised for my well-being
even at this moment of time

ii
To,
The Controller of Examinations,
University of Veterinary and Animal Sciences,
Lahore.

We, the supervisory committee, certify that the contents and form of the thesis
submitted by Farzana Abbas, Reg. No. 2007-VA-532 have been found satisfactory
and recommend that it be processed for evaluation, by External Examiner(s) for the
award of degree.

SUPERVISORY COMMITTEE:

Chairman : ------------------------------------------------
(Prof. Dr. Muhammad Ashraf)

Member : --------------------------------------------------
(Dr. Sumaira Abbas)

Member : ---------------------------------------------------
(Dr. Arshad Javid)

iii
ACKNOWLEDGEMENTS

Praises are all attributed to ALMIGHTY ALLAH, the omnipotent and sovereign; blessed and
bestowed me with the power of pen and knowledge, mightier than the sword, and raised my
humble self to a status he choose for His loved ones, namely those who know. Sure, the teaching
and preaching of HOLY PROPHET HAZRAT MUHAMMAD (SAW), the torch of guidance,
source of knowledge and wisdom for mankind, have ever been an inspiration for getting on with
unknown and to invade the land of erudition.
How could I, all alone, be thrilled to achieve this task, the recognition certainly goes to my
seniors, specially my kind supervisor, Prof. Dr. Muhammad Ashraf, Chairman Department of
Fisheries and Aquaculture, University of Veterinary and Animal Sciences, Lahore, Pakistan. His
intellectual support, expert advice and constant encouragement developed my confidence and
pursued me for writing of this dissertation.
I am indebted to Dr. Sumaira Abbas, Assistant Professor, Department of Fisheries and
Aquaculture, Dr. Arshad Javid, Assistant Professor, Department of Wildlife and Ecology
University of Veterinary and Animal Sciences, Lahore, Pakistan for their friendly advices and
prompt assistance during the study.
A special thanks, goes without saying, to my class fellows; Mr. Khalid Javed Iqbal, Mrs.
Sydah Andleeb, Mr. Iftikhar Ahmed Qureshi, Mrs. Ghina Islam, Ms. Rida Younis and field
workers for their intimate support and warm appreciations.
Family members are true assets of love and prayers in the back. It gives me pleasure to extend
my gratitude to my friendly husband, Dr. Muhammad Hafeez-ur-Rehman, who has been a
constant and invariable source of moral as well as academic support, making me more and more
proud of him.
I am delighted to thank my father and mother whom hands are always raised for my successful
career and my sisters and brothers for their untiring efforts to support me to my satisfaction.
Their patience and deep prayers are always with me in every field of life.
May ALLAH bless them all with long and peaceful lives. Amin!
Farzana Abbas

iv
TABLE OF CONTENTS

Page
Chapter Contents
No.
Acknowledgement iv
Table of Contents v
List of Tables ix
List of Figures xi
Abstract xii
Chapter 1
1 1
General Introduction
Chapter 2
2 7
Review of Literature
Lernaea, its prevalence, infestation and susceptibility
7
Body composition
11
Hematological studies
14
Immunity studies
16
Literature cited
21
Chapter 3
Experiment 1.
3 37
L. cyprinacea Susceptibility, Infestation and its Treatment in Indigenous
Major and Exotic Chinese Carps under Polyculture System.
Introduction 37
Materials and Methods 40
Experimental site 40
Experimental design 40
Parasite collection, preservation and identification 40
Growth parameters 41
Water quality parameters 41
Pond treatment 41
Statistical analysis 43
Results 43
v
Parasite identification 43
Weight and length gain in control and treated group 44
Infestation of L. cyprinacea during the course of study in control and treated
44
groups
Discussion 50
Literature Cited 54
Chapter 4
Experiment-2
4 60
Comparison of biochemical profile of healthy and infectious Chinese,
Indian and Exotic carps.
Introduction 60
Materials and Methods 64
Experimental site 64
Experimental design 64
Water quality parameters 66
Proximate Analysis 66
Moisture and Dry Matter Test 66
Ash contents 67
Crude protein 67
Crude fat (Ether extract) 69
Mineral analysis 69
Wet digestion of samples 69
Determination of Sodium (Na) and Potassium (K) by flame photometer 70
Determination of Phosphorus (P) by spectrophotometry 70
Determination of Iron (Fe), Zinc (Zn), Calcium (Ca), Copper (Cu) and
71
Magnesium (Mg)
Statistical analysis 71
Results 72
Bio-chemical analysis 72
Mineral analysis 74
Water quality parameters 76
Discussion 78
vi
Literature Cited 84
Chapter 5
Experiment-3
5 93
Comparison of Various Blood Indices among Chinese and Indian Major
Carps From the Perspectives of Their Resistance Against Lernaeaosis
Introduction 93
Materials and Methods 97
Experimental site 97
Experimental Design 97
Blood haematology 97
Blood chemistry 99
Ig M determination 99
Statistical analysis 99
Result 100
Blood hematology 100
Blood chemistry 107
Discussion 110
Literature Cited 115
Chapter 6
6 Experiment-4 126
Fish Mucus; A Physical Barrier to Pathogens
Introduction 126
Materials and Methods 128
Experimental site 128
Experimental animal 128
Experimental procedure 128
Mucus collection 128
Total Protein Contents 129
Polyacrylamide Gel Electrophoresis (SDS-PAGE) 129
Lectin activity 129
Alkaline Phosphatase Test 130
vii
Statistical analysis 130
Results 130
Protein concentration 130
Protein profile of mucus 131
Lectin activity 132
Alkaline Phosphatase Test 133
Discussion 134
Literature Cited 138
Chapter 7
Experiment 5
7 Essential and Non-essential Amino Acid Composition of Indian and 144
Chinese Major Carps from the Perspective of Fish Immunity against
Lernaeosis
Introduction 144
Materials and Methods 146
Experimental site 146
Experimental animal 146
Parasitic infestation 146
Amino acid analysis 146
Statistical analysis 147
Results 147
Parasitic prevalence 147
Amino acid profiling 149
Discussion 152
Literature Cited 155
Collective Summary 160

viii
LIST OF TABLES
Table Page
Title
No. No.
Morphometric measurements of experimental fish species at the initiation of
1 42
trial.
2 Feed Formula; ingredient and nutrient composition 42
3 Weight and length gain in the control and treated group during the study period. 45
Average number of L. cyprinacea on different body parts during study period in
4 46
control group. (Mean ± SD)
Average number of L. cyprinacea on different body parts during study period in
5 47
treated group. (Mean ± SD)
Number of L. cyprinacea and its %age on different body parts of experimental
6 47
fish in control ponds (without treatment of DDVP) during the course of study.
Number of L. cyprinacea and its %age on different body parts of experimental
7 48
fish in treated ponds (DDVP) during the course of study.
8 Mean values for water quality parameters during the course of study 48
Pearson correlation among various water quality parameters (weekly basis)
9 49
during the course of study
Pearson correlation among various water quality parameters (fortnightly basis)
10 49
during the study period.
Morphometric measurements of experimental fish species at the initiation of
1 65
trial.
2 Feed Formula used during the course of studies 65
3 Proximate Analysis of fishes from treated group 72
Proximate Analysis of fishes from control group with heavy L. cyprinacea
4 73
infestation
5 Comparison of macronutrients in fishes from treated and control group 73
6 Mineral contents of fishes from treated group 74
7 Mineral contents of fishes from control group infected with L. cyprinacea 75
8 Comparison of minerals (ppm) concentrations in treated and control group 76
9 Mean values for Water quality parameters during the course of study 77
Pearson correlation between the parameters (weekly basis) during the course of
10 77
study
Pearson correlation between the parameters (fortnightly basis) during the study
11 78
period.
Comparison of blood parameters related to red blood cells among experimental
1 100
fish species
2 Comparison of differential leukocyte count in experimental fish species 101
3 Comparison of less important blood parameters in experimental fish species 102
ix
4 Comparison of blood parameters of experimental fish species females 103
5 Comparison of blood parameters between males of experimental fish species. 104
6 Comparison of blood parameters of males and females of Indian major carps 105
7 Comparison of blood parameters of males and females of Chinese carps 106
8 Total protein, albumin and globulin in females of experimental fish species 108
9 Total protein, albumin and globulin in males of experimental fish species 108
Total protein, albumin and globulin between genders of experimental fish
10 108
species
11 Total protein, albumin and globulin between species of experimental fish species 109
Total protein, albumin and globulin between males and females of experimental
12 109
fish species
Protein concentration, HA titer and alkaline phosphatase activity in mucus of
1 131
experimental fish species
Molecular weight of protein found in the skin mucus of experimental fish
2 132
species (SDS-PAGE analysis)
Number of L. cyprinacae and its %age on different body parts of experimental
1 148
fish in treated ponds (DDVP) during the course of study
Essential amino acid composition of the whole body tissues of experimental fish
2 150
species
Non-essential amino acid composition of the whole body tissues of experimental
3 151
fish species

x