SHYNE ANAND PS

251 Pages·2012·7.08 MB·English

EFFECTS OF DIETARY SUPPLEMENTATION BY
PERIPHYTON BIOMASS AND BIOFLOC ON GROWTH
AND IMMUNO-PHYSIOLOGICAL ASPECTS OF JUVENILE
PENAEUS MONODON FABRICIUS, 1798

Thesis submitted in partial fulfillment
of the requirements for the degree of

Ph.D. (Aquaculture)
By

SHYNE ANAND P.S.
(Ph.D. 260)

CENTRAL INSTITUTE OF FISHERIES EDUCATION
(University under Section 3 of UGC act)
Indian Council of Agricultural Research
Versova, Mumbai- 400 061
SEPTEMBER 2012
Dedicated to ……….

My beloved daughter
Pragya
DECLARATION

I hereby declare that the thesis entitled “EFFECTS OF DIETARY
SUPPLEMENTATION BY PERIPHYTON BIOMASS AND BIOFLOC
ON GROWTH AND IMMUNO-PHYSIOLOGICAL ASPECTS OF
JUVENILE PENAEUS MONODON FABRICIUS, 1798” is an authentic
record of the work done by me and that no part thereof has been
presented for the award of any degree, diploma, associateship,
fellowship or any other similar titles. I accept the guidelines of ICAR for
IPR related issues.

(SHYNE ANAND P. S.)
Date:
PhD Scholar
Place: Mumbai, India Aquaculture
ACKNOWLEDGEMENT
This dissertation would not have been possible without the guidance and
the help of several individuals who in one way or another contributed and
extended their valuable assistance in the preparation and completion of this
study.
First and foremost, I would like to express my deep and sincere gratitude
to my supervisor, Dr. M. P. Singh Kohli, Principal scientist (Retd.), Division of
Aquaculture, CIFE, for his guidance, scholarly advice, constant encouragement
and critical evaluation during the entire duration of my Ph.D work.
With proud regard, I express my deep sense of gratitude to Dr. W. S.
Lakra, Director, CIFE for providing me all the necessary facilities needed for
the successful completion of My Ph.D work.
It is my proud privilege to express my deep sense of gratitude to Dr. A.
G. Ponniah, Director, Central Institute of Brackishwater Aquaculture, Chennai
for his generosity, encouragement and granting me study leave and providing me
all the necessary facilities required to conduct my research work to
successfully finish this Ph.D programme.
I am deeply indebted to Dr. S. Dam Roy, Director, CARI, Port Blair and
my advisory member for his guidance and kind consideration regarding my
academic requirements throughout my research period.
I express my sincere gratitude to Dr. J. K. Sundaray, Officer-In charge,
Kakdwip research centre of CIBA and advisory member for guidance and
providing me all the necessary facilities to conduct my Ph.D. work.
I wish to express my warm and sincere thanks to Dr. Archana Sinha,
Officer-In charge, Kolkata Centre of CIFRI and Dr. G. H. Pailan, my advisory
committee members for their timely help, guidance and support during my
research work.
A great deal of thanks goes to Dr. N. K. Chadha, principal scientist, Dr. A.
K. Pal, Joint Director, DR. Neelam Saharan, HOD, Aquaculture, Dr. Kiran Dube
principal scientist, Dr. A. K. Jaiswar, Senior scientist, Dr. Chandra Prakash,
Senior scientist, Dr. Munil Kumar Sukham, Senior scientist, Dr. V. K. Tiwari,
Senior scientist, Dr. A. K. Verma (SS) and Dr. Paromitha, scientist, CIFE for
providing me valuable suggestions, timely support and academic help during the
study period. I wish to express my sincere thanks to all scientists and staffs of
CIFE for their direct or indirect help.
I am deeply indebted to Dr. C. S. Purushothaman, HOD, Aquatic
environment management division, Dr. G. Venkateshwarlu, HOD, Post harvest
division for their necessary assistance and allowing me to analyse sample in their
lab, and the help rendered by Mr. A. K. Padmanaban (Technical Officer), Aquatic
environment management division and Mr. Janardanan, Ph.D scholar, CIFE in
analyzing the sample is sincerely acknowledged.
With proud regard, I express my sincere gratitude to Dr. P.
Ravichandran, HOD Crustacean culture Division, CIBA, Dr. A. R.T. Arasu, Fish
culture Division, CIBA, Dr. S. M. Pillai, Principal scientist, CIBA for the moral
support, advice and necessary assistance during my research work. I am
indebted to all scientists and staffs of CIBA, for allowing me to access all the
facilities.
I am indebted to Dr. T. K. Ghoshal, Dr. D. De and Dr. A. D. Deo, senior
scientist of Kakdwip Research Centre CIBA, for their valuable suggestions,
support and allowing me to access the laboratory facilities.
I am grateful to scientists at CIFRI, Barrackpore Dr. Mohanty, Dr. Amiya Sahoo
for their timely help and, allowing me to access the laboratory facilities.
I express my special thanks to Deepa , Pavan, Sajana, Soma, Madona,
Prabitha, Murali, Biji, Babi, Sipra, Nagesh, Datta and my affectionate juniors
Reshma, Manju, Ratheesh, Ranjith, Arun ,Chanu for their love, affection and
making my life at CIFE colourful. I wish to thank all my batchmates, juniors and
fellow Ph.D students at CIFE for all their support and timely help.
A heartfelt thanks to technical staffs at Kakdwip Research Centre,
Ramakrishna, Soma, Dr. S. B. Bhatacharya, Pintu, Vimal, Pravanjan and Amar da
for assisting me in laboratory and field work. I thank my friends in Kakdwip,
West Bengal and elsewhere for their support and encouragement throughout my
study period
I sincerely acknowledge the ICAR that provided the necessary financial
support for this research.
No words can express my genuine thanks to my dear parents, Amma,
Achan, Pitaji and Maa who were a source of constant love and inspiration
throughout my life. My sister Sajini Chechi, Ragini didi and brother Vishnu and
Ajit have given me their unequivocal support throughout, as always, for which
my mere expression of thanks does not suffice.
I owe my most sincere gratitude and deeply indebted to Biva di for her
kindness and support in taking care of my little daughter and patience which
helped to keep me in a good state of mind during the study period.
Above all, I wish to record my deep feelings of affection and loving
thanks to my husband Dr. Sujeet Kumar for his great patience, constructive
criticism in thesis writing, moral support during difficult times and the untiring
effort at each and every step to make this a success.
I owe my deep love and endless obligations to my little daughter, Pragya
who missed her motherly love many times due to my busy work schedule and
even suffered burn injury during my Ph.D. research period. I dedicate this piece
of work to my little daughter.
Lastly, I bow my head before almighty the omnipresent God, for answering my
prayers and whose blessings have contributed all success till this point in my
life.

Shyne Anand P. S.
ABSTRACT
Two separate growth trials were conducted to evaluate the graded levels of
periphyton and biofloc in the diet of Penaeus monodon juveniles for their growth,
immune response, digestive and metabolic enzyme activity. Periphyton biomass
was produced in outdoor cistern tanks (15 m2) over a period of 45 days using
bamboo substrates. Biofloc was generated in indoor tanks (1,000 L) at C: N ratio 10
using wheat flour and ammonium sulphate as carbohydrate and nitrogen source. In
growth trial 1, a sixty day feeding trial was conducted using juvenile P. monodon
(2.20 ± 0.04 g) in indoor FRP tanks (1,000 L) with graded level of periphyton
biomass inclusion in shrimp basal diets; 0% (control) , 3% (T1); 6% (T2); 9% (T3),
and a positive control with natural bamboo substrate for periphyton growth (T4). At
the end of the experiment, significantly better growth performance (p<0.01) in terms
of specific growth rate (SGR), feed conversion ratio (FCR), protein efficiency ratio
(PER) and, significantly higher (p<0.01) digestive enzyme activities were noticed in
T2 and T4 compared to other treatments. The level of metabolic enzymes like
lactate dehydrogenase (LDH), malate dehydrogenase (MDH), aspartate
aminotransferase (AST) and alanine amino transferase (ALT) were lower in
treatments compared with control. The better immune response (p<0.05) in terms of
total haemocyte count (THC), superoxide dismutase (SOD), catalase, serum protein
and, disease resistance against Vibrio harveyi was noticed in T2 compared with
control. Similarly in trial 2, a sixty day feeding trial was conducted, using juvenile P.
monodon (2.90 ± 0.10 g) in indoor FRP tanks (1000 L) with graded level of dried
biofloc inclusion in shrimp basal diets; 0% (control), 4% (T1); 8% (T2) and 12% (T3).
At the end of feeding trial, T1 and T2 showed significantly better (p<0.05) growth
performance and digestive enzyme activities compared with control. Level of
metabolic enzymes like LDH, MDH, AST and ALT was significantly lower in
treatments compared with control. Biofloc incorporated diet significantly elevated the
immune response and disease resistance against V. harveyi compared with control.
This suite of experiment demonstrated the dietary supplementation at 6 and 4%
level of periphyton and biofloc respectively found to be effective in growth
improvement and to elicit immuno-physiological response in tiger shrimp Penaeus
monodon juvenile.
CONTENTS
SL.
Title Page No.
NO.
1. INTRODUCTION 1-3
2. REVIEW OF LITERATURE 4-28
2.1. Microorganism in Aquaculture 4
2.2. Periphyton 4
2.2.1 Periphyton dynamics and influencing factors 5
2.2.2 Periphyton biomass 7
2.2.3 Taxonomic diversity of periphyton 9
2.2.4 Nutritional composition of periphyton 10
2.2.5 Role of periphyton in aquaculture 11
2.3. Biofloc Technology 13
2.3.1 Factors influencing biofloc production 14
2.3.2 Microbial community in biofloc 15
2.3.3 Nutritional composition of biofloc 16
2.3.4 Role of biofloc in aquaculture 17
2.3.5 Biofloc as dietary ingredient 18
2.3.6 Biofloc as dietary stimulant 19
2.3.7 Biofloc in shrimp health 19
2.4 Crustacean Physiology 20
2.4.1 Digestive enzymes 20
2.4.2 Microbial role in digestive enzyme activities 22
2.4.3 Enzymes of phosphomonoesterase 23
2.4.4 Enzymes in protein metabolism 23
2.4.5 Enzymes in pentose phosphate pathway 24
2.4.6 Enzymes in carbohydrate metabolism 25
2.4.7 Enzymes in Kreb’s cycle 25
2.5. Shrimp Immune System 26
2.5.1 Haemocytes 26
2.5.2 Antioxidant enzymes 27
2.5.3 Haemolymph metabolites 28
3. MATERIALS AND METHODS 29-66
3.1 Experiment I 29
3.1.1 Site of experiment 29
3.1.2 Periphyton production 29
3.1.3 Physicochemical parameters of water 34
3.1.4 Periphyton biomass estimation 34
3.1.5 Taxonomic analysis of periphyton 35
3.1.6 Proximate analysis of periphyton 36
3.1.7 Mineral composition of periphyton 38
3.1.8 Fatty acid profile of periphyton 39
3.2 Experiment II 39
3.2.1 Site of experiment 39
3.2.2 Chemicals and glassware 40
3.2.3 Experimental animals 40
3.2.4 Experimental diets 40
3.2.5 Experimental design and set-up 42
3.2.6 Physico-chemical parameters of water 43
3.2.7. Proximate analysis of experimental diets 43
3.2.8. Growth performance 44