In the name of Allah, Most Gracious, Most Merciful! STUDIES ON THE EFFECT OF WOOD EXTRACTIVES IN COMBINATION WITH PLANT OIL ON SUBTERRANEAN TERMITES By BABAR HASSAN M.Sc. (Hons.) Entomology 2006-ag-1738 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY IN ENTOMOLOGY DEPARTMENT OF ENTOMOLOGY FACULTY OF AGRICULTURE UNIVERSITY OF AGRICULTURE, FAISALABAD PAKISTAN 2017 And when We decreed (Solomon's) death, they had no indication that he was dead until (they saw a termite), a crawler of the earth eating away his staff. And when he fell down, the jinn realized that had they known the unseen, they would not have continued in their humiliating punishment Al-Quran (XXXIV. 13) Declaration I hereby declare that the contents of the thesis, " Studies on the effect of wood extractives in combination with plant oil on subterranean termites" are product of my own research and no part has been copied from any published source (Except the references, standard mathematical or generic models/ formulates/protocols etc.). I further declare that this work has not been submitted for award of any other diploma/degree. The university may take action if the information provided is found incorrect at any stage. In case of any default the scholar will be proceeded against as per HEC policy. Babar Hassan 2006-ag-1738 DEDICATED TO MY SWEET LATE MOTHER AND MY LOVING FATHER Whose love is more precious than pearls and diamonds, by the virtue of whose prays, I have been able to reach at this high position ACKNOWLEDGEMENTS Alhamdullilah, all praises to Allah for the strengths and His blessings in completing this thesis. All praises be to the Holy Prophet Muhammad (Peace be upon him), the city of knowledge, the illuminating torch and the rescuer of humanity from going astray. Without the help of the people, to whom I wish to express my gratitude this thesis could not be completed. First and foremost, I would like to thank Prof. Dr. Sohail Ahmed, Department of Entomology University of Agriculture, Faisalabad, for his sympathetic attitudes, fatherly behavior, animate directions, observant pursuit, scholarly criticism, cheering perspective and enlightened supervision. Dr. Muhammad Altaf Sabri and Prof. Dr. Shahbaz Talib Sahi, as member of supervisory committee are acknowledged for their helpful advice and suggestions. I would also like to express my special appreciation to foreign advisors, Carol Clausen, Mark, E. Mankowski and Grant Kirker, USDA-FS, Forest Products Laboratory (USA). They have been a tremendous mentors for me. I would like to thank all of them for encouraging during research and for allowing me to grow as a research scientist. Their advices on both the research as well as on learning career have been priceless. I am also thankful to Dr. Muhammad Misbah-ul-Haq (NIFA) and Dr. Khalid Zamir Rasib (FC College University) for providing me space for laboratory and field experiments against Heterotermes indicola. I would like to acknowledge and thank Higher Education Commission of Pakistan (HEC) and USDA- FS International Program, who granted me fellowship and PhD Indigenous Scholarship to conduct research at USDA-FS, Forest Products Laboratory (USA) and the Nuclear Institute for Food and Agriculture (NIFA), Peshawar (Pakistan). I am also thankful to Craig Bell, USFS FPL Starkville, MS for wood block and shaving preparation, Amy Bishell for decay tests and Dr. Hamid Borazjani, Mississippi State University, for use of his laboratory facilities. My deepest gratitude goes to my family for their unflagging love and support throughout my life; this dissertation is simply impossible without them. I want to acknowledge my brothers Anser Abbas and Qaiser Abbas and my sisters for their love, sincerity and cheerful jokes, which make possible my stay here in this University. My gratitude will remain incomplete if I do not mention the contribution of the sincere friends, Rahat Afza, Asad Ahmad, Basit Naseer, Nadeem Ahmad, Tahmmal Huassain, Ali Anwar and laboratory fellows, Ali, Nasir, Awias, Muzammal, Bilal and Uzair Saleem for their great company and advices. Thanks to all. BABAR HASSAN CONTENTS CHAPTER TITLE PAGES ACKNOWLEDGMENTS vii CONTENTS viii LIST OF TABLES ix LIST OF FIGURES xiii LIST OF APPENDICES xiv ABSTRACT xv 01 INTRODUCTION 01 02 REVIEW OF LITERATURE 07 03 MATERIALS & METHODS 27 04 RESULTS 49 05 DISCUSSIONS 145 06 SUMMARY 159 LITERATURE CITED 163 APPENDICES 194 LIST OF TABLES Sr. No. Title of the Table Page No. 3.1. Properties of durable woods selected for current study 29 3.2. Indicators to Evaluate Antifeedant Activity of extractives (Antifeedancy %) 33 3.3. Termite rating scheme (AWPA E-1, E-7, E-26) 47 3.4. Decay rating scheme (AWPA E-10, E-7) 48 4.1. Yield of heartwood extractives from durable woods extracted with ethanol: 50 toluene (2:1) 4.2.1. Median lethal concentrations (LC50s) of filter paper treated with four type of 52 heartwood extractives against R. flavipes and H. indicola 4.2.2. Mean weight loss, area loss and mortality of R. flavipes and H. indicola after 53 toxicity test on filter paper treated with T. grandis heartwood extractives 4.2.3. Mean weight loss, area loss and mortality of R. flavipes and H. indicola after 54 toxicity tests on filter paper treated with D. sissoo heartwood extractives 4.2.4. Mean weight loss, area loss and mortality of R. flavipes and H. indicola after 55 toxicity tests on filter paper treated with C. deodara heartwood extractives 4.2.5. Mean weight loss, area loss and mortality of R. flavipes and H. indicola after 56 toxicity tests on filter paper treated with P. roxburghii heartwood extractives 4.2.6. Mean mortality (%) of R. flavipes and H. indicola after toxicity tests on 57 filter paper treated with four heartwood extractives. 4.2.7. Mean weight loss (%) of filter papers after feeding of R. flavipes and H. 57 indicola treated with four heartwood extractives 4.2.8. Mean area loss (%) of filter papers after feeding of R. flavipes and treated 58 with four heartwood extractives 4.2.9. Antifeedant activity of T. grandis heart wood extractives against R. flavipes 67 and H. indicola 4.2.10. Antifeedant activity of D. sissoo heart wood extractives against R. flavipes 67 and H. indicola 4.2.11. Antifeedant activity of C. deodara heartwood extractives against R. flavipes 68 and H. indicola 4.2.12. Antifeedant activity of P. roxburghii heartwood extractives against R. 68 flavipes and H. indicola 4.2.13. Antifeedant activity of four heartwood extractives against R. flavipes and H. 69 indicola 4.2.14. Mean GSTs activity of H. indicola exposed to IC50s concentrations of BHT, 72 quercetin and heartwood extractives 4.2.15. Mean ESTs activity of H. indicola exposed to IC50s concentrations of BHT, 72 quercetin and heartwood extractives 4.2.16. Mean CATs activity of H. indicola exposed to IC50s concentrations of BHT, 72 quercetin and heartwood extractives 4.3.1 Effect of T. grandis heartwood extractives on total population of gut 75 protozoans of R. flavipes and H. indicola 4.3.2. Effect of D. sissoo heartwood extractives on total population of gut 76 protozoans of R. flavipes and H. indicola 4.3.3. Effect of C. deodara heartwood extractives on total population of gut 77 protozoans of R. flavipes and H. indicola 4.3.4. Effect of P. roxburghii heartwood extractives on total population of gut 78 protozoans of R. flavipes and H. indicola 4.3.5. Percentage reduction of total population of gut protozoans of R. flavipes and 79 H. indicola after feeding on filter paper treated with four durable heartwood extractives 4.3.6. Number of sequences, OTUs, and percent coverage for each sample by 81 treatment 4.3.7. Sequence similarity analysis of isolated bacterial strains from H. indicola 83 with the available databases in NCBI 4.4.1. Mean weight loss (%) for extractive free and un-extracted blocks of T. 86 grandis and mortality of R. flavipes and H. indicola under Choice and No- choice tests 4.4.2. Mean weight loss (%) for extractive free and un-extracted blocks of D. 86 sissoo and mortality of R. flavipes and H. indicola under Choice and No- choice tests 4.4.3. Mean weight loss (%) for extractive free and un-extracted blocks of C. 88 deodara and mortality of R. flavipes and H. indicola under Choice and No- choice tests 4.4.4. Mean weight loss (%) for extractive free and un-extracted blocks of P. 88 roxburghii and mortality of R. flavipes and H. indicola under choice and no- choice tests 4.4.5. Average rating, retention, mortality and weight losses of SYP and CW 90 treated with T. grandis extractives after feeding of R. flavipes 4.4.6. Average rating, retention, mortality and weight losses of SYP and CW 92 treated with D. sissoo extractives after feeding of R. flavipes 4.4.7. Average rating, retention, mortality and weight losses of SYP and CW 93 treated with C. deodara extractives after feeding of R. flavipes 4.4.8. Average rating, retention, mortality and weight losses of SYP and CW 94 treated with P. roxburghii extractives after feeding of R. flavipes 4.4.9. Average rating, retention, mortality and weight losses of SYP and CW 96 treated with T. grandis extractives after feeding of H. indicola 4.4.10. Average rating, retention, mortality and weight losses of SYP and CW 97 treated with D. sissoo extractives after feeding of H. indicola 4.4.11. Average rating, retention, mortality and weight losses of SYP and CW 99 treated with C. deodara extractives after feeding of H. indicola 4.4.12. Average rating, retention, mortality and weight losses of SYP and CW 100 treated with P.