1 ECOLOGY AND MANAGEMENT OF TERMITES IN URBAN ENVIRONMENT (ISLAMABAD) ABDUL SATTAR 08-arid-772 Department of Entomology, Faculty of Crop and Food Sciences Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi Pakistan 2015 ECOLOGY AND MANAGEMENT OF TERMITES IN URBAN ENVIRONMENT (ISLAMABAD) by 2 ABDUL SATTAR (08-arid-772) A thesis submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Entomology Department of Entomology, Faculty of Crop and Food Sciences Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi Pakistan 2015 CERTIFICATION I hereby undertake that this research is an original one and no part of this thesis falls under plagiarism. If found otherwise, at any stage, I will be responsible for the consequences. Student’s Name: ABDUL SATTAR Signature: ______________________ Registration No.: 08-arid-772 Date: __________________________ Certified that the contents and form of the thesis entitled “Ecology and Management of Termites in Urban Environment (Islamabad)” submitted by Mr. Abdul Sattar have been found satisfactory for the requirements of the degree. 3 Supervisor: __________________________ (Dr. Muhammad Naeem) Co-Supervisor: _______________________ (Dr. Ehsan-ul-Haq) Member:_____________________________ (Dr. Ata-ul-Mohsin) Member:_____________________________ (Dr. Fayyaz-ul-Hassan) Chairman: ___________________ Dean: _______________________ Director, Advanced Studies: ____________________________ 4 IN THE NAME OF ALLAH THE MOST MERCIFUL THE MOST GRACIOUS DEDICATION 5 I dedicate this humble effort to my parents, whose unprecedented love and affection can never be compensated Abdul Sattar CONTENTS Page List of Tables xi List of Figures xiii Acknowledgments xvi Abstract xviii 1 INTRODUCTION 1 2 REVIEW OF LITERATURE 6 2.1. ECOLOGICAL STUDIES 6 2.1.1. Population dynamics 7 2.1.2. Foraging ecology 9 2.1.3. Species identification 10 2.1.4. Caste composition in termite colony 11 2.2. EVALUATION OF DYE MARKERS 12 2.3. PLANTS AND THEIR PRODUCTS 14 2.4. PHAGOSTIMULANTS 15 i) Yeast 16 6 ii) Urea 16 iii) Plant Extracts 17 iv) Glucose 17 v) Chemicals 18 2.5. BAITS 19 i) Metabolic inhibitors 20 ii) Fungi (bioagent) 21 iii) Insect Growth Regulators (IGRs) 21 3 IMPACT OF ENVIRONMENTAL FACTORS ON THE 22 POPULATION DYNAMICS, DENSITY AND FORAGING ACTIVITIES OF O. LOKANANDI AND M. OBESI IN ISLAMABAD 3.1. ABSTRACT 22 3.2. INTRODUCTION 23 3.3. MATERIALS AND METHODS 25 3.3.1 Ecological Studies 25 3.3.2. Survey 26 3.3.3. Population Dynamics of Subterranean Termites 26 3.3.4. Identification of termites 26 3.3.5. Ecology of foraging termites 27 3.3.6. Statistical Analysis 27 3.4. RESULTS 27 3.4.1. Ecological studies of termites in urban environment 27 (Islamabad) 3.4.1.1. Survey 27 3.4.1.2. Population dynamics 28 3.4.1.3. Species of termites in the experimental areas 29 3.4.1.4. Yield (g) and number of termites in 1.0 gm sample 29 3.4.1.5. Foraging Ecology of Subterranean Termites. 33 3.4.1.6. Caste composition of foraging groups of subterranean 36 termites. 3.5. DISCUSSION 37 3.5.1. Population dynamics 37 3.5.2. Foraging Ecology 41 4 EFFECT OF DYE-MARKERS I.E. NILE BLUE-A AND SUDAN 44 RED-7B ON MICROTERMES OBESI AND ODONTOTERMES LOKANANDI 4.1. ABSTRACT 44 4.2. INTRODUCTION 44 4.3. MATERIALS AND METHODS 46 4.3.1. Biological Stains 46 4.3.2. Experimental Termites 47 4.3.3. Preparation of Dye Attractive Bait 47 4.4. RESULTS 49 7 4.4.1. Toxicity of Biological dyes 49 4.4.2. Nile blue-A 49 4.4.3. Sudan red-7B 53 4.5. DISCUSSION 59 5 INVESTIGATION OF DYE MARKERS WHICH REMAIN 62 VISIBLE IN BODY OF TERMITES AT DIFFERENT RELATIVE HUMIDITIES 5.1. ABSTRACT 62 5.2. INTRODUCTION 63 5.3. MATERIALS AND METHODS 64 5.3.1. Experimental Termites 64 5.3.2. Visibility of dye markers in the body of termites under at 65 different relative humidity 5.3.3. Retention test 66 5.4. RESULTS 66 5.4.1. Visibility of dye markers i.e., Nile blue-A and Sudan 66 red-7B in the body of termites at different relative humidities 5.4.1.1. Nile blue-A 66 5.4.1.2. Sudan red-7B 69 5.4.2. Retention of dye 69 5.5. DISCUSSION 70 6 SCREENING OF PLANT EXTRACTS TO FIND OUT 82 PROPER CONCENTRATION FOR DEVELOPMENT OF SLOW-ACTING TOXICANT BAITS TO MANAGE TERMITES 6.1. ABSTRACT 82 6.2. INTRODUCTION 82 6.3. MATERIALS AND METHODS 84 6.3.1. Collection of Experimental Termites 84 6.3.2. Plants Collection and their Extracts Preparation 85 6.3.3. Bioassay 85 6.3.3.1. Toxicity test 85 6.3.4. Statistical Analysis 86 6.4. RESULTS 86 6.4.1. Microtermes obesi 86 i) Euphorbia helioscopia (leaf extracts) 87 ii) Euphorbia helioscopia (seed extracts) 87 6.4.2. Odontotermes lokanandi 89 8 i) Euphorbia helioscopia (leaf extracts) 89 ii) Euphorbia helioscopia (Seed extracts) 90 6.4.3. Microtermes obesi 91 i) Cannabis sativa (leaf extracts) 91 ii) Cannabis sativa (Seed extracts) 95 6.4.4. Odontotermes lokanandi 96 i) Cannabis sativa (Leaf extracts) 96 ii) Cannabis sativa (Seed extracts) 97 6.4.5. Microtermes obesi 99 i) Calotropis procera (Leaf extracts) 99 ii) Calotropis procera (Seed extracts) 101 6.4.6. Odontontermes lokanandi 103 i) Calotropis procera (Leaf extracts) 103 ii) Calotropis procera (Seed extracts) 105 6.5. DISCUSSION 106 7 LABORATORY INVESTIGATION OF COPER 111 SULPHATE AND MERCURIC CHLORIDE TO FIND OUT PROPER CONCENTRATION TO BE USED IN SLOWACTING TOXICANT BAITS FOR MANAGEMENT OF TERMITES 7.1. ABSTRACT 111 7.2. INTRODUCTION 111 7.3. MATERIALS AND METHODS 113 7.3.1. Studies on the Efficacy of Mercuric Chloride and Copper 113 Sulphate 7.3.2. Bioassay 113 7.3.2.1. Toxicity Test 113 7.3.3. Statistical Analysis 114 7.4. RESULTS 114 7.4.1. Efficacy of Copper Sulphate and Mercuric Chloride 114 against Microtermes obesi 7.4.1.1. Toxicity Test 114 7.4.2. Efficacy of Copper Sulphate and Mercuric Chloride 115 against Odontotermes lokanandi 7.4.2.1. Toxicity Test 115 7.5. DISCUSSION 116 9 8 SCREENING OF DIFFERENT COMPOUNDS TO FIND OUT 121 PHAGOSTIMULANT TO MAKE ATTRACTIVE BAIT FOR THE CONTROL OF SUBTERRANEAN TERMITES 8.1. ABSTRACT 121 8.2. INTRODUCTION 121 8.3. MATERIALS AND METHODS 123 8.3.1. Studies of different compounds to find out 123 Phagostimulants 8.3.1.1. Determination of Phagostimulant as potential bait 123 substrates for Microtermes obesi 8.3.2. Preparation of Poplar saw Dust Extract 124 8.3.2.1. Determination of poplar saw dust extract as potential 124 bait substrates for Microtermes obesi 8.3.3. Comparative attractancy Test 125 8.4. RESULTS 126 8.4.1. Eualvation of different compounds to find out 126 phagostimulants i) Effect of different compounds (urea, yeast and 126 glucose) on bait consumption and survival of Microtermes obesi ii) Effect of different concentrations of poplar sawdust 127 extract on bait consumption and survival of Microtermes obesi 8.4.2. Comparative attractancy test 1: Distilled water, 0.1% 131 urea, poplar sawdust extract, 3% glucose, and 3% yeast. 8.4.3. Comparative attractancy test 2: Distilled water, 0.1% 131 urea,poplar sawdust extract, 3% glucose, and 4% yeast 8.4.4. Comparative attractancy test 3: Distilled water, 0.1% 131 urea, poplar sawdust extract, 4% glucose, and 2% yeast 8.4.5. Comparative attractancy test 4: Distilled water, 1% urea, 133 poplar sawdust extract, 2% glucose, and 1% yeast 8.4.6. Comparative attractancy test 5: Distilled water, 1% urea, 133 poplar sawdust extract, 1% glucose, and 1% yeast 8.4.7. Comparative attractancy test 6: Distilled water, 1% urea, 133 4% yeast, 4% glucose, and poplar sawdust individually and in different combinations 8.5. DISCUSSION 134 9 FORMULATION OF SLOW-ACTING TOXIC BAITS TO 142 CONTROL SUBTERRANEAN TERMITES 9.1. ABSTRACT 142 9.2. INTRODUCTION 142 9.3. MATERIALS AND METHODS 144 9.3.1. Formulation of Slow-acting Toxicant Baits 144 9.3.1.1. Experimental Termites 144 9.3.1.2. Choice Feeding Test 145 9.4. RESULTS 146 10 9.4.1. Formulation of Palatable toxicant baits for Microtermes 146 obesi by combining of phagostimulants with different toxicants 9.4.2. Comparative percent bait consumption by M. obesi 155 9.5. DISCUSSION 156 10 GENERAL DISCUSSION 162 SUMMARY 178 RECOMMENDATIONS 183 LITERATURE CITED 184 APPENDICES 256 LIST OF TABLES Table No. Page 3.1 Infestation of NIFA-TERMAPs by O. lokanandi (O) and M. 32 obesi (M) at Islamabad during 2010 to 2012 3.2 Mean yield (g), mean number of termites in 1.0 gm sample and 34 mean percent workers in foraging group of M. obesi collected from “NIFA TERMAPs” installed in Islamabad from September 2010 to September 2012 3.3 Mean yield (g) and mean number of termites in 1.0 gm sample 35 and mean percent workers in foraging group of O. lokanandi collected from “NIFA TERMAPs” installed in Islamabad from September 2010 to September 2012 11 3.4 Correlation between environmental factors and biomass of 39 termite species captured through “NIFA TERMAPs” from Islamabad 3.5 Correlation between environmental factors (Atmospheric 39 Temperature, Relative Humidity and Precipitation) and % workers of Termites 6.1 Mean percent mortality in M. obesi at different concentrations 92 of leave and seed extracts of E. helioscopia 6.2 Mean percent mortality in O. lokanandi at different 93 concentrations of leave and seed extracts of E. helioscopia 6.3 Mean percent mortality in M. obesi at different concentrations 98 of leave and seed extracts of C.