Table Of ContentEvaluation of Stress Reducing Capacity of Selected
Anaesthetics for the Live Transportation of
Green Chromide Etroplus suratensis
Thesis submitted to
Cochin University of Science and Technology
in partial fulfillment of the requirements
for the degree of
Doctor of Philosophy
in
Fisheries Management
Under the Faculty of Marine Sciences
by
Sindhu M.C
SCHOOL OF INDUSTRIAL FISHERIES
COCHIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
KOCHI – 682016
February, 2015
Evaluation of Stress Reducing Capacity of Selected Anaesthetics for
the Live Transportation of Green Chromide Etroplus suratensis
Ph.D. Thesis under the Faculty of Marine Sciences
Author
Sindhu M.C
Research Scholar
School of Industrial Fisheries
Cochin University of Science and Technology
Kochi - 682016
Email:
Dedicated to
The Small Scale Fish Farmers of Kerala
SCHOOL OF INDUSTRIAL FISHERIES
COCHIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
KOCHI – 682016
Dr. A. Ramachandran Ph: 9447062400
Professor email:
SCHOOL OF INDUSTRIAL FISHERIES
COCHIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
KOCHI – 682016
Dr. A. Ramachandran Ph: 9447062400
Professor email:
I, SINDHU M.C, do hereby declare that the present work entitled
“Evaluation of Stress Reducing Capacity of Selected Anaesthetics for
the Live Transportation of Green Chromide Etroplus suratensis” is an
authentic work done by me under the guidance and supervision of
Dr. A. Ramachandran, Professor and supervising guide, School of Industrial
Fisheries, Cochin University of Science and Technology, Kochi-16. The
work presented in the thesis has not been presented for any other degree or
diploma earlier.
Sindhu M.C
Kochi - 682016
February, 2015
At the outset I wish to thank God, Almighty for his grace and blessing for
making this work a successful one.
I would like to express my sincere gratitude to my supervisor, Professor Dr.A
Ramachandran, who deserves thanks for his contribution, positive attitude, new
ideas and guidance.
I would like to thank my research committee members, Dr. Saleena Mathew,
Dr. Harikrishnan for their guidance throughout my work.
I also wish to thank our Director, Professor Dr.K.T .Thomson, and Dr.Mini
Sekharan for their guidance.
I would also like to thank Dr. Sudhir, Professor and Head, Department of
Ayurveda college, Government of Kerala, Thripunithura.India and Dr. Anil at
Government Ayurveda College, Puthiyakavu; Thripunithura, for helping me to study
the possible application of plant anesthetics as a tool for transportation of live fishes.
I am also very thankful for the guidance and selfless invaluable analytical
assistance of Professor Angel Mathew (Research and Analytical wing UGC), at the
Great Maharajas College who also provided selfless invaluable assistance with
experimental design and statistical analysis towards completion of the thesis.
Of course, none of this would have been fulfill without the tireless support of
Dr. M.Mukundan, the former Director, CIFT, INDIA and Dr.K.S. Purushan,
former Dean, College of Fisheries, Panangad, Kochi,… whose are the grate
passionate for aquaculture and provided critical reading of the manuscripts, which I
am very grateful for.
Thanks to Dr. Sasi Menon, Sajil, Sinu, Sini, Soumya, at the Faculty of
Biochemistry, DDC International, Kochi for assistance with blood sampling and
analysis.
At KUFOS, I am very grateful to Dr. Manjusha who selflessly gave her time,
advice, and support over the years which helped me immensely to finish this thesis
promptly.
Also, I am grateful to the library in charges of the School of Marine Sciences
at the University of Cochin, Mr.Manuel for their constant encouragement,
especially, Thadevus sir is also an ornamental fish hobbyist and culturist, who give
practical ideas in shaping my research design.
I am indebted to Marson, Aqua farm Puthuvyppu, Fish farmer and exporter who
supplied the plenty of juveniles Etroplus suratensis, for the entire length of my study. It
would have been impossible for me to complete the work without his support.
I am grateful to my colleagues Soumya Subrade, Liya Jayalal, Antony, Asha,
Anupama, Anjaly, Dayana were always there to help me when ever I was in need. I
thank them wholeheartedly.
I sincerely thank staff, School of Industrial Fisheries for their timely helps.
I record my thanks to Mr. Binoop (Indu photos & Graphics) for his perfect sense
of commitment which helped a great deal in the layout and design aspect of my thesis.
My brothers Biju and Shaiju were always keen about my research and I thank
them for their curiosity.
I owe my father (Late), mother and mother in law for their love, support and
confidence throughout the past years.
I am deeply obliged to Nandu and Kunju for their understanding, who were
adjust with the thick and thin research days of their Amma.
Last, but certainly not least, I thank my family for their unfailing support and
love throughout this journey, especially my husband, Gopinath without whose love,
encouragement and understanding, this work would never have been materialized!
And after everything else but not least, all the little fish that gave (?) their life
to science—thank you!!
Sindhu M.C
General Introduction ......................................................... 01 - 27
a). Kerala and State Fish “Karimeen” (Etroplus suratensis) ...................... 16
b). Transportation of juveniles of Etroplus suratensis .......................... 22
c). Justification .................................................................................... 24
d). Scope of the study .......................................................................... 25
e). Objectives of the study .................................................................. 26
Review of Literature .......................................................... 29 - 47
a). Transportation of Fishes ........................................................................ 29
b). Water Quality ................................................................................ 30
c). Stress ............................................................................................. 33
d). Scope of the study .......................................................................... 35
Chapter 1
Acute toxicity of selected anaesthetics on
juveniles of Etroplus suratensis correlates with
certain water quality parameters .................................... 49 - 154
1.1 Introduction ................................................................................... 49
1.2 Materials and Methods ................................................................... 53
1.2.1 Juveniles of Etroplus suratensis (Bloch 1790) ............................... 53
1.3 General protocol ............................................................................ 55
1.3.1 Specimen collection and acclimatization ........................................ 55
1.4 Anaesthetic agents ......................................................................... 56
1.4.1 Preparation of anaesthetics agents ................................................. 56
1.4.1.1 Preparation of clove oil (Syzygium aromaticum) ...................... 56
1.4.1.2 Preparation of cinnamon oil (Cinnamomum zeylanicum) ............. 57
1.4.1.3 Preparation of Cassumunar ginger (Zingiber cassumunar
Roxb)......................................................................................... 58
1.4.1.4 Preparation of Tobacco leaf extracts (Nicotiana tobaccum)........... 59
1.4.1.5 Preparation of Tricaine methanesulfonate (MS-222) .................... 60
1.4.1.6 Preparation of Hypothermic condition ......................................... 61
1.5 Experimental set up for anaesthetization of fish.............................. 63
1.5.1 Experimental design ...................................................................... 63
1.5.1.1 Determination of acute toxicity (96 h LC50) test .......................... 63
1.5.1.2 Mortality change in pattern of the fish during 24, 48, 72,
and 96 h .................................................................................... 64
1.5.1.3 Determination of water quality parameters ................................... 65
1.5.2 Post treatment survival .................................................................. 65
1.6 Statistical analyses ......................................................................... 66
1.7 Results ........................................................................................... 66
1.7.1 Effect of clove oil .......................................................................... 67
1.7.2 Effect of Cinnamon oil .................................................................. 74
1.7.3 Effect of Zingiber cassumunar Roxb ............................................. 81
1.7.4 Effect of Tobacco leaves extract (Nicotiana tobacum) ................... 87
1.7.5 Effect of MS-222 (Tricaine methanesulfonate) .............................. 94
1.7.6 Effect of Hypothermic condition.................................................. 100
1.8 Discussion .................................................................................... 106
1.8.1 Effect of clove oil ........................................................................ 106
1.8.2 Effect of Cinnamon oil ................................................................ 115
1.8.3 Effect of Cassumunar ginger extracts (Zingiber cassumunar Roxb) .. 122
1.8.4 Tobacco leaves extract (Nicotiana tobacum) ............................... 128
1.8.5 Effect of MS-222 ........................................................................ 137
1.8.6 Effect of Hypothermic condition.................................................. 143
1.9 Summary ...................................................................................... 149
Chapter 2
Behavioural assays and safest level of selected
anaesthetics during bath administration in
different concentrations on juveniles of Etroplus
suratensis ......................................................................... 155 - 233
2.1 Introduction .................................................................................. 155
2.2 Materials and Methods .................................................................. 166
2.2.1 Fish and experimental conditions .................................................. 166
2.2.2 Experiments ................................................................................. 166
2.2.2.1 Effect of ethanol as an anaesthetic. ............................................. 166
2.2.2.2 Investigation of the safest level and behavioral assays
during the exposure of different concentration of clove oil,
cinnamon oil, cassumunar ginger extract, tobacco leaf
extract, MS222 and hypothermia .............................................. 167
2.2.2.3 Video Recording System ........................................................... 169
2.2.3 Post treatment survival ............................................................................ 170
2.3 Statistical analyses ........................................................................ 170
2.4 Results .......................................................................................... 170
2.4.1 Effect of Ethanol as an anaesthetic............................................... 170
2.4.2 Effect of Clove oil as an anaesthetic ............................................ 171
