POPULATION GENETIC STRUCTURE OF INDIGENOUS ORNAMENTAL TELEOSTS, PUNTIUS DENISONII AND PUNTIUS CHALAKKUDIENSIS FROM THE WESTERN GHATS, INDIA Thesis submitted in partial fulfillment of the requirement for the Degree of Doctor of Philosophy in Marine Sciences of the Cochin University of Science and Technology Cochin – 682 022, Kerala, India by LIJO JOHN (Reg. No. 3100) National Bureau of Fish Genetic Resources Cochin Unit CENTRAL MARINE FISHERIES RESEARCH INSTITUTE (Indian Council of Agricultural Research) P.B. No. 1603, Kochi – 682 018, Kerala, India. December, 2009. Declaration I do hereby declare that the thesis entitled “Population genetic structure of indigenous ornamental teleosts, Puntius denisonii and Puntius chalakkudiensis from the Western Ghats, India” is the authentic and bonafide record of the research work carried out by me under the guidance of Dr. A. Gopalakrishnan, Principal Scientist and SIC, National Bureau of Fish Genetic Resources (NBFGR) Cochin Unit, Central Marine Fisheries Research Institute, Cochin in partial fulfillment for the award of Ph.D. degree under the Faculty of Marine Sciences of Cochin University of Science and Technology, Cochin and no part thereof has been previously formed the basis for the award of any degree, diploma, associateship, fellowship or other similar titles or recognition. Cochin (Lijo John) 16th December 2009 ®É¹]ÅÒªÉ ¨ÉiºªÉ +ÉxÉÖÖ´ÉÆÆζÉE ºÉÆƺÉÉvÉxÉ ¤ªÉÚ®Éä NATIONAL BUREAU OF FISH GENETIC RESOURCES NBFGR Cochin Unit, CMFRI Campus, P.B. No. 1603, Cochin-682 018, Kerala, India Fax: (0484) 2395570; E-mail: [email protected] Dr. A. Gopalakrishnan, Date: 16.12.2009 Principal Scientist, Officer-in-Charge & Supervising Teacher Certificate This is to certify that this thesis entitled, “Population genetic structure of indigenous ornamental teleosts, Puntius denisonii and Puntius chalakkudiensis from the Western Ghats, India” is an authentic record of original and bonafide research work carried out by Mr. Lijo John (Reg. No. 3100) at National Bureau of Fish Genetic Resources (NBFGR) Cochin Unit, Central Marine Fisheries Research Institute, under my supervision and guidance in partial fulfilment of the requirement for the award of the degree of Doctor of Philosophy in the Faculty of Marine Sciences, Cochin University of Science and Technology, Cochin, Kerala. It is also certified that no part of the work presented in this thesis has been submitted earlier for the award of any degree, diploma or any other similar title. (A. Gopalakrishnan) ACKNOWLEDGEMENTS First of all, I bow before the God almighty and praise him for the gracious love and blessing showered on me. I express my sincere gratitude to Dr. A. Gopalakrishnan, (supervising guide), Principal Scientist and Officer-in-Charge, National Bureau of Fish Genetic Resources (NBFGR) Cochin Unit, for his unstinted support and valuable guidance throughout the course of study. I am extremely thankful and indebted to Dr. E.G. Silas, (former Vice Chancellor of Kerala Agriculture University and former Director of CMFRI), for his valuable advices and critical suggestions which helped me much during several critical stages of the course of study, without which this research work would not have been so nicely accomplished.. It’s my pleasure, to thank Dr. K.K. Vijayan, Principal Scientist and Head, Marine Biotechnology Division, CMFRI, for his timely help, moral support and encouragement extended during the final stages of my research and thesis preparation. I sincerely express my gratefulness to Dr. W.S. Lakra, Director of NBFGR, for providing all facilities at NBFGR Cochin Unit to work for my Ph.D. thesis and Dr. G. Syda Rao Director of CMFRI, Cochin and Prof. (Dr.) Mohan Joseph Modayil, former Director of CMFRI, for making room for my Ph.D. registration under Cochin University of Science and Technology (CUSAT) and for the promptness in taking all necessary actions for the smooth functioning of the research programme. I am indebted to Dr. P.C. Thomas, Principal Scientist and Officer-in-Charge of HRD Cell, CMFRI, for his cooperation, valuable advice, and timely help during my Ph.D. programme. I take this opportunity to thank Dr. A.V. Saramma, Professor, School of Marine Sciences, Cochin University of Science and Technology for sparing some of her valuable time as the Expert member of my Doctoral Research Committee. I also thank the concerned authorities of CUSAT for giving me the registration for Ph.D. and for further proceedings. I am extremely thankful to Dr. Alex Eapen (Scientist C, National Institute of Malaria Research, Chennai) and Dr. Rema Devi K. (Scientist E, Marine Biology Research Centre, Zoological Survey of India, Chennai) for their valuable suggestions and comments during the course of my investigation and at final stages of my thesis preparation. With great respect and regards, I acknowledge Drs. Kuldeep Kumar Lal and Vindhya Mohindra, Principal Scientists, NBFGR, Lucknow, for the help rendered by them on various practical aspects especially in the beginning stages of my research. I am also thankful to Drs. T. V. Anna Mercy and Dinesh Kaipilly (College of Fisheries, Panangadu, Kochi) for their help during the initial stages of my Ph.D. programme especially for sampling. I express my sincere gratitude to Dr. V.S. Basheer, (Sr. Scientist), Dr. Divya, P.R. Dr. Rajaswaminathan, (Scientists of NBFGR Cochin Unit) and Dr. Karan Veer Singh (Scientist, NBFGR,, Lucknow), for their encouragement throughout my Ph.D. work. I am also thankful to Dr. P. Vijayagopal (Senior Scientist, CMFRI, Kochi) for the help rendered during final stages of my Ph.D. programme. I iv feel happy to express my thanks to Mr. M. P. Paulton (Sr. Technical Assistant, CMFRI, Cochin) and Mr. Nandakumar (Technical Assistant, CMFRI) for their timely help during the period of Ph.D. work. I would also like to thank Dr. Manoj, P. (Sr. Technical Assistant, RGCB, Thiruvanandapuram) for his help in sequencing DNA samples. I feel happy to express my thanks to my colleagues at NBFGR Cochin Unit, Mr. Alfred Dourom, Siby Philip, Ms. Jeena, N.S., Vidya, R,, Princy Thomas and Sajeela, K.A., for their help and support. I express my sincere thanks to my friends Mr. Reynold Peter, Anoop, B., Jeswin Joseph, Ranjith Kumar, Vikas, P.A., Sajesh Kumar, Shiju, P., (Research Scholars of Marine Biotechnology Division, CMFRI), Tiju Jacob, Rishad, K.S., Shine Kumar, Allen Benziger, and Thangaraj, M., for all the encouragement and moral support given by them. I also thank my seniors, Drs. Abdul Muneer, P.M., Musammilu, K.K. (NBFGR Cochin Unit) and Jyothi. V. Mallia (CMFRI), for the encouragement and help rendered by them during the initial stages of my research. Mr. Ratheeshan, K and Ambrose, T.V. (Research Scholars of Demersal Fisheries Division, CMFRI) wholeheartedly supported me in carrying out the statistical analysis of morphometric data, my sincere thanks to them for the help and support. My sincere thanks to Mr. Shankaran (Artist, CMFRI), for sketching the map of Kerala that has been included in the thesis. For the moral support extended to me, the Scientists of NBFGR and CMFRI and the remaining vast circle of my friends, including my batch mates, lab mates, seniors and juniors for Ph.D. are affectionately remembered. My teachers (graduation level), Dr. Mathew Joseph, Shri. Francis Joseph, Dr. M.J. Paul, and Shri. Earney E. Paul, were always there to encourage me; my sincere thanks to them for their encouragement and moral support. I am grateful to OIC library, other staff members in the library, staff members of HRD Cell, canteen, security and other members of CMFRI, Cochin, for their sincere help and cooperation extended during the course of my study. I sincerely acknowledge the Research Fellowship from the Ministry of Earth Sciences (MoES), which I have received while pursuing the Ph.D. course. My special thanks are due to Mr. Babu T.L., Mr. Sithosh, Mr. Ratheesh, Mr. Boney, Mr. Trijo, Mr. Saby (Fish Hobbyists), Mr. Johney and Mr. Baby (local fishermen) who where wholeheartedly assisted and helped me in the field at different locations for collecting fish specimens during my field trips. Finally, I owe much more than I can to my parents, C.A John my father, Laly John my mother and all my family members, whose prayers, blessings, sincere and constant support have enabled me to bring out this piece of work. To them I dedicate this thesis. (Lijo John) v Preface Our riverine system, especially the torrential streams and rivers originating in the Western Ghats are known to have variety of ornamental fishes. Except for traditional taxonomic description we have little information on these species, their ecology, biology, life habits, life history and population structure. There is large scale depletion of habitats due to anthropogenic factors which affect the ecosystems of these streams. At the same time, a growing trade in ornamental fish resulting in the exploitation of the resources from the wild stock threatening their existence. On account of this, Government of Kerala State has promulgated a ban on the capture from wild of one of the species Puntius denisonii (Day) during its peek breeding season. The importance of learning more about the species such as P. denisonii and its associated species has motivated me to take-up the present study, the subject of this thesis entitled “Population genetic structure of indigenous ornamental teleosts, Puntius denisonii and Puntius chalakkudiensis from the Western Ghats, India.” My study has brought out some important findings on P. denisonii species complex which I am sure will help towards developing management strategies for conservation and well being of the species. The thesis is presented in five chapters excluding summary and conclusion and references. I have adopted updated techniques both in analyzing meristic and morphometric data and genetic data relating to mitochondrial DNA to arrive at conclusions. This work has carried out under the guidance of Dr.