AAPPPPLLIICCAATTIIOONN OOFF BBIIOOFFLLOOCC TTEECCHHNNOOLLOOGGYY ((BBFFTT)) IINN TTHHEE NNUURRSSEERRYY RREEAARRIINNGG AANNDD FFAARRMMIINNGG OOFF GGIIAANNTT FFRREESSHHWWAATTEERR PPRRAAWWNN,, MMAACCRROOBBRRAACCHHIIUUMM RROOSSEENNBBEERRGGIIII ((DDEE MMAANN)) Thesis submitted to the CCoocchhiinn UUnniivveerrssiittyy ooff SScciieennccee aanndd TTeecchhnnoollooggyy in partial fulfillment of the requirements for the degree of DDooccttoorr ooff PPhhiilloossoopphhyy in AAqquuaaccuullttuurree UUnnddeerr tthhee FFaaccuullttyy ooff MMaarriinnee SScciieenncceess by PPRRAAJJIITTHH KK..KK..,, MM..SScc..,, AARRSS Reg No: 3564 SCHOOL OF INDUSTRIAL FISHERIES COCHIN UNIVERSITY OF SCIENCE AND TECHNOLOGY KOCHI – 682016 DECEMBER 2011 AApppplliiccaattiioonn ooff BBiioofflloocc TTeecchhnnoollooggyy ((BBFFTT)) iinn tthhee nnuurrsseerryy rreeaarriinngg aanndd ffaarrmmiinngg ooff ggiiaanntt ffrreesshhwwaatteerr pprraawwnn,, MMaaccrroobbrraacchhiiuumm rroosseennbbeerrggiiii ((ddee MMaann)) Ph.D. Thesis under the Faculty of Marine Sciences     Author Prajith K.K Research Scholar School of Industrial Fisheries Cochin University of Science and Technology Kochi - 682016 Email: [email protected] Supervising Guide Prof. B. Madhusoodana Kurup. PhD Vice-Chancellor Kerala University of Fisheries & Ocean Studies Panangad, Kochi- 682 506 Email: [email protected] School of Industrial Fisheries Cochin University of Science and Technology Kochi - 682016 December 2011 Front cover Prawn consuming biofloc developed in the experimental tanks Back Cover Biofloc formed in the experimental tanks DDeeddiiccaatteedd ttoo…… AAllll aanniimmaallss ggrroowwnn iinn mmyy eexxppeerriimmeennttaall ttaannkkss bbyy ccoonnssuummiinngg bbiioofflloocc && TToo tthhee ffaarrmmeerrss wwhhoo pprraaccttiiccee ssuussttaaiinnaabbllee aaqquuaaccuullttuurree KKEERRAALLAA UUNNIIVVEERRSSIITTYY OOFF FFIISSHHEERRIIEESS && OOCCEEAANN SSTTUUDDIIEESS (Established on 20th day of November 2010 and governed by the Kerala University of Fisheries and Ocean Studies Act. 2010 passed by the Kerala Law (Legislation 1) Department vide notification No. 19540/Leg.1/2010/Law dated 28th January 2011) Prof. B. Madhusoodana Kurup. PhD Panangad, Kochi – 682 506, Kerala, India Phone:0484 2703781 (Direct), 2700598, 2700964 Post Doc. (Wageningen University, The Netherlands) (Res) 0484- 2703944, 2700424 Fax: (91) (O) 484 2700337 Vice-Chancellor E-mail: [email protected], [email protected] [email protected] This is to certify that the thesis entitled “Application of Biofloc Technology (BFT) in the nursery rearing and farming of the giant freshwater prawn, Macrobrachium rosenbergii (de Man)” to be submitted by Mr. Prajith K. K, is an authentic record of research work carried out by him under my guidance and supervision in partial fulfillment of the requirement of the degree of Doctor of Philosophy in Aquaculture of Cochin University of Science and Technology, under the faculty of Marine Sciences. Prof. (Dr.) B. Madhusoodana Kurup (Supervising Guide) Kochi - 682016 December 2011 DDeeccllaarraattiioonn  I, Prajith K.K, do hereby declare that the thesis entitled “Application of Biofloc Technology (BFT) in the nursery rearing and farming of Giant freshwater prawn, Macrobrachium rosenbergii (de Man)” is a genuine record of research work done by me under the supervision of Prof. (Dr.) B. Madhusoodana Kurup, Vice-Chancellor, Kerala University of Fisheries and Ocean Studies, Panangad, Kerala, India and has not been previously formed the basis for the award of any degree, diploma, associate- ship, fellowship or other similar title of any university or institution. Kochi – 682016 Prajith K.K December 2011 I take this special moment to express my sincere thanks and deepest sense of gratitude to my guide Prof. (Dr.) B. Madhusoodana Kurup, Vice Chancellor, Kerala University of Fisheries and Ocean Studies, Cochin for his unfailing guidance, invaluable suggestions, critical assessment and constant encouragement throughout my work. I gratefully acknowledge the financial support by the European Commission under the 6th frame work programme on “Environmental management reform for the sustainable farming, fisheries and aquaculture- AquAgris” for my research. Thanks to the Honorable President, ICAR for granting extension of time to join Agricultural Research Service for completing my thesis. I always remember the support, infrastructure and facilities offered by the former Directors of the School of Industrial Fisheries, Cochin University of Science and Technology during my work period. Thanks to Prof. (Dr.) A. Ramachandran, Director of School of Industrial Fisheries for providing necessary facilities to carry out this work successfully. Thanks are due to Prof. (Dr.) Ramakrishnan Korakandy, Prof. (Dr.) Saleena Mathew, Prof. (Dr.) K.T. Thomson, Dr. M. Harikrishnan, Dr. Mini Shekaran, Dr. John Mohan, for the support and wishes. Thanks to Prof (Dr) Aneykutty Joseph, Dr. Rosamma Philip, Prof. (Dr.) Babu Philip, Dr. A. A. Mohammend Hatha, Dr. A.V. Saramma and Dr. S. Bijoy Nandan, my post graduation teachers from Dept of Marine biology Microbiology and Biochemistry, CUSAT for their support and wishes for the timely completion of the thesis. I am deeply acknowledging Prof. Yoram Avnimelech, (Israel) the “Father of Biofloc Aquaculture” who was ready to help me at any time. Thanks to my graduation teachers, Dr. Baby Joseph, Dr. Joselet Mathew, Prof. Jancy George and Dr. Sally George, for their affection, care and support. A relation of 7 long years has presented many memorable moments with my class mates, Akhilesh, Vinu, Shuaib, Jithu, Chaithanya, Manju, Jimly, Ramya, Bisna, Anu Baburaj, Anu Pavithran, Divya, Abhijna, Asha, Sumi, Renuka and Archana, who always supported and criticized during this work. It’s my luck to get the truly co-operative research group and a good research environment. I thank Joyni Thomas for sharing her knowledge and offering help during the project and research period, thanks to Anjalee Devi for providing me literature, rendering her whole hearted help and well wishes. Dr. Saritha Thomas, Shubhasree Shankar, just like my elder sisters, thanks for the care and love. Mr. Ranjith Kumar. C. R, Mrs. Diana Benjamin, Mrs. Anupama, Mr.Jenson. V. R, Mr. Deepak Jose, Mrs. Roshni and Miss. Greeshmam were always with me as inspiration in my academic achievements. Special thanks to Dr. Prabhakaran M. P. Dept of Marine Biology, Microbiology and Biochemistry, CUSAT, for taking up the hectic task of correcting the entire thesis. Thanks are due to Dr. C. Sreedevi, Lecturer, SIF, CUSAT for the initial correction of the thesis. Help offered by the office staff of School of Industrial fisheries and Library Staff of School of Marine Sciences is thankfully acknowledged. MSc students of various batches helped me a lot in each and every stage of my research for the seed collection harvest, sample analysis etc. Thanks to Mr. Sajan K.P, Shenoob P. S, Anil J, Nithin C.T, Praveen P, Judin John Chako, Ajith, Lijin Nambiar, Rahul, Mahesh Gopal and Vijeesh, I am fortunate for the friendship of Mr. Thushar Patel, Gujarat, for helping me a lot for the proper reviewing of my work by providing latest and recent articles in the field of BFT. Miss. Anuradha, Senior Research Scholar, Dept. of Chemical Oceanography helped me a lot from the beginning of the research by providing and sharing the methodology and profound knowledge in water Chemistry. Special thanks to Mr. Krishna Mohan, for providing essential software for research and sharing his computer knowledge. Thanks to Dr.Suneesh Thampy and Mr.Rahul, MPEDA for providing statistical data related to this work. Thanks to the scholars of Department of Marine Biology, especially Miss. Chaithanaya, for assisting me in the analysis of molecular biology samples. Thanks are due to Mr. Anil Kumar. P. R, Mr. Anit. M.Thomas, Dr. Sanil Kumar, for sharing their knowledge in the relevant field and helping by providing their lab facility. Special thanks to Mr.Binoop, Indu photos, Kalamassery for the perfect editing of this text. Mr. Shuaib, my friend designed a beautiful and attractive cover page to this manuscript. For the last 7 years, each and every moment in the CUMS hostel was enjoyed very much, thanks to all my hostel friends, for those gorgeous days, Jayesh, Anilkumar, Shaiju, Phiros, Vijay, Anit, Gireesh, Vijayakumar, Haseeb, Sudheer Mathew, Jabir, Ajin, Vincent, Christo, Vipin….there are so many names. I can’t forget the sweet memories in Room No 5, with my roommates, Subin, Nikhil, Midhun and finally my dearest Riyas, Solly and Nashad. Thanks to all for the love and support. Achan, Amma, Aniyathy and Maamy are my strength and have unconditionally supported me with their love, care, prayers and blessings. No words to suffice my feeling for them, at this critical juncture in my life. This thesis is a result of blessing and kind of Lord Guruvayoorappan, I bow my head in front of that super power, which preserves the entire world and showers the power and blessings to all in this beautiful world. Chapter 1 INTRODUCTION...............................................................................................01 - 22 1.1 Introduction-------------------------------------------------------------01 1.2 World aquaculture-----------------------------------------------------02 1.3 Indian aquaculture: Its growth and development -------------03 1.4 Mariculture -------------------------------------------------------------04 1.5 Brackishwater aquaculture------------------------------------------05 1.6 Freshwater aquaculture ----------------------------------------------06 1.7 Giant freshwater prawn, Macrobrachium rosenbergii as the candidate species for freshwater aquaculture in India-------08 1.8 Status of giant freshwater prawn culture-------------------------09 1.9 Biofloc Technology (BFT) and its application for sustainable aquaculture ---------------------------------------------------------------13 1.10 Status of biofloc aquaculture----------------------------------------19 1.11 Hypothesis, objective and outline of the thesis ------------------21 Chapter 2 REVIEW OF LITERATURE................................................................................23 - 42 2.1 Sustainable aquaculture----------------------------------------------23 2.2 Environmental problems of aquaculture -------------------------24 2.3 Concept of biofloc technology and its application in aquaculture systems as a tool for waste management----------28 2.4 Application of biofloc technology in giant freshwater prawn aquaculture-----------------------------------------------------39          Chapter 3 APPLICATION OF BIOFLOC TECHNOLOGY IN THE PRIMARY NURSERY REARING OF GIANT FRESHWATER PRAWN, MACROBRACHIUM ROSENBERGII, IN VARYING STOCKING DENSITY: EFFECT ON ANIMAL WELFARE AND SEED QUALITY..................43 - 66 3.1 Introduction-------------------------------------------------------------43 3.2 Materials and methods------------------------------------------------45 3.2.1 Tank allocation----------------------------------------------------45 3.2.2 Larval maintenance and assessment----------------------------47 3.2.3 Water quality -----------------------------------------------------48 3.2.4 Statistical analysis ------------------------------------------------48 3.3 Results--------------------------------------------------------------------49 3.4 Discussion----------------------------------------------------------------56 3.5 Conclusion---------------------------------------------------------------66 Chapter 4 APPLICATION OF BIOFLOC TECHNOLOGY IN THE GROW-OUT SYSTEM OF GIANT FRESHWATER PRAWN, MACROBRACHIUM ROSENBERGII: EFFECT ON WATER QUALITY AND PRODUCTION......................................................67 - 94   4.1 Introduction-------------------------------------------------------------67 4.2 Materials and methods------------------------------------------------70 4.2.1 Tank facilities and design----------------------------------------70 4.2.2 Calculation of quantity of carbohydrate required for biofloculation----------------------------------------------------------72 4.2.3 Assessment of water and sediment quality parameter --------73 4.2.4 Shrimp yield parameters-----------------------------------------74 4.2.5 Statistical analysis ------------------------------------------------75 4.3 Results --------------------------------------------------------------------75 4.3.1 Water and sediment quality--------------------------------------75 4.3.2 Prawn yield parameters ------------------------------------------81 4.4 Discussion----------------------------------------------------------------83 4.4.1 Efficiency of BFT in the grow-out system of giant freshwater prawn and optimization of protein in the feed-----------------------------------------------------------------83 4.4.2 Comparison of the production of giant freshwater prawn in BFT applied and normal culture systems -----------87 4.5 Conclusion---------------------------------------------------------------94 Chapter 5 EFFICACY OF VARIOUS CARBOHYDRATE SOURCES AS BIOFLOCULATING AGENT IN THE GROW-OUT SYSTEM OF GIANT FRESHWATER PRAWN, MACROBRACHIUM ROSENBERGII..........................................................................95 - 118   5.1 Introduction-------------------------------------------------------------95