VIETNAM NATIONAL UNIVERSITY OF AGRICUTURE FACULTY OF AGRONOMY UNDERGRADUATE THESIS TITLE: “STUDYING ON SOME BIOLOGICAL CHARACTERISTICS OF TETRAGONULA IRIDIPENNIS AND TETRAGONULA FUSCOBALTEATA CAMERON 1908 (APIDAE: MELIPONA) IN GIA LAM, HANOI 2020” Student : Le Thanh Luong Code : 611619 Class : K61KHCTT Supervisor : Assoc Prof Dr Pham Hong Thai Hanoi,2021 ii COMMITMENT I pledge that the contents of this report are honestly conducted by me under the guidance of Assoc Prof Dr Pham Hong Thai I hereby undertake that all references in this document are quoted which are the name of the authors, name of researches, time, location, and orgins I take full responsibility for the report Hanoi, March 3, 2021 Student Le Thanh Luong i ACKNOWLEDGEMENT In order to complete the graduation thesis, in addition to my own efforts, I have received the enthusiastic help and guidance from teachers, and the care of family and friends First of all, I would like to express my sincere gratitude to Assoc Prof Dr Pham Hong Thai, Department of Entomology, Vietnam National University of Agriculture directly instructed me wholeheartedly and helped me in the process of completing and completing the thesis I would like to express my sincere thanks to all the teachers of the Department of Insect, Agronomy of Faculty, Vietnam National University of Agriculture for their interest, creating the best conditions for me to implement the topic I would like to express my sincere thanks to all starch of Research Center for Tropical Bees and Beekeeping, Vietnam National University of Agriculture directly helped and supported me in the thesis Finally, I would like to thank my family and friends who have always supported and encouraged me throughout the learning and implementation process ii ABLES OF CONTENT COMMITMENT i ACKNOWLEDGEMENT ii ABLES OF CONTENT .iii LIST OF TABLE v LIST OF FIGURE vi ACRONYMS viii ABSTRACT .ix PART I: INTRODUCTION 1.1 General information 1.2 Objectives and Requirements 1.2.1 Objectives 1.2.2 Requirements PART II: LITERATURE REVIEW 2.1 Taxonomy .3 2.1.1 Taxomony .3 2.1.2 Stingless bee species recorded in Asia 2.2 Biological characteristics of stingless bees .9 2.2.1 Caste of stingless bees on colony 2.2.2 Life cycle .11 2.2.3 Behaviors .11 PART III: MATERIALS AND METHODS .19 3.1 Objects and materials 19 3.1.1 Objects 19 3.1.2 Materials 19 3.1.3 Time and research location 19 3.2 Research Contents 19 iii 3.3 Methodology 20 3.3.1 Identifying and preserving stingless bee specimens .20 3.3.2 Studying on some biological characteristics of T iridipennis and T fuscobalteata 21 PART IV: RESULTS AND DISCUSSION 24 4.1 Diversity of stingless bee species in Vietnam .24 4.1.1 Number of Subgenera and Species in some provinces in Vietnam: .24 4.1.2 Morphological characteristics for identification stingless bee species in experimental locations: .26 4.2 Biological characteristics of T iridipennis Smith 1854 and T fuscobalteata Cameron 1908 36 4.2.1 Worker brood cells 36 4.2.2 Oviposition behaviors 38 PART V: CONCLUSION AND RECOMMENDATION 50 5.1 Conclusions 50 5.2 Recommendation 51 REFERENCES 52 APPENDIX .56 iv LIST OF TABLE Table 4.1 Composition of stingless bee species in four provinces in Vietnam, 2020 .24 Table 4.2 Subgenera of stingless bee species were found 25 Table 4.3 Worker cell dimensions 36 Table 4.4.Cell capacity and Brood-food volume of worker brood-food 37 Table 4.5 Ratio of brood-food 38 Table 4.6 The average hour of cell-building by workers 39 Table 4.7 The average of food-discharge duration of workers (seconds) 43 Table 4.8 The average number of workers who inserted during provisioning phase 44 Table 4.9 Inserted duration of the ten first workers 44 Table 4.10 Egg-laying duration of queen (seconds) 46 Table 4.11 Egg Dimension .47 Table 12 Operculation time (minutes) and number of workers operculated 49 v LIST OF FIGURE Figure 4.1 Mesoscutum of specimen 27 Figure 4.2 Frontal face of specimen 27 Figure 4.3 Hamuli in wing of specimen 27 Figure 4.4 Frontal face of specimen 28 Figure 4.5 Bare spot in propodeum of specimen 28 Figure 4.6 Mesoscutum of specimen 29 Figure 4.7 Hamuli in wing of specimen 29 Figure 4.8 Mesoscutum of specimen 30 Figure 4.9 Hair arrangement on mesoscutum of T iridipennis (Sakagami, 1978) .30 Figure 4.10.Frontal face of specimen 30 Figure 4.11 Hamuli in wing of specimen 30 Figure 12 Mesoscutum of specimen .32 Figure 13 Metasomal tergum in Lepidotrigona ventralis, Sakagami 1975 32 Figure 4.14.Hamuli in wing of specimen 32 Figure 4.15.Fontal face of specimen 32 Figure 16 Frontal face and forewing of specimen 33 Figure 17 Hindleg of specimen 33 Figure 18 Mesoscutum of specimen .33 Figure 19 Lateral view of specimen .33 Figure 4.21.Frontal face of specimen 34 Figure 4.20 Facial view and Forewing major veins of Lisotrigona carpenter Engel (Engel et al, 2018) 34 Figure 4.22 Frontal face of specimen .35 Figure 4.23 Frontal face Homotrigona fimbriata by Siti et al 35 Figure 4.27 Mesoscutum of specimen 35 vi Figure 4.26 Lateral view of specimen 35 Figure 4.25 Abdomen of specimen 35 Figure 4.24 Hamuli of specimen 35 Figure 4.28 The mean height of growing cell related to that of egg-containing cell .40 Figure 4.29 The number of cells which were building at started stage 41 Figure 4.30 The percentage of cell building stage at am .42 Figure 4.31 Inserted duration of the ten first workers 45 Figure 4.32 1-day-old egg 47 vii ACRONYMS RCTB : Research Center for Tropical Bees and Beekeeping DVE : Digital Video Eyepiece viii PART V: CONCLUSION AND RECOMMENDATION 5.1 Conclusions - There are nine species with six subgenera of one genus were identified: Homotrigona fimbriata Smith 1857, Lepidotrigona ventralis Smith 1857, Lisotrigona carpenter Engel 2000, Tetragonula fuscobalteata Cameron 1908, Tetragonula iridipennis Smith 1854, Tetragonula laviecep Smith 1857, Tetrigonilla collina Smith 185, Tetrigona apicalis Smith 1857, Lepidotrigona terminate Smith 1878 There is one unknown species, it belongs to sub genus Tetragonula - The worker brood cell dimension and cell volume of T iridipennis and T fuscobalteata are significantly different It was determined: 3.07mm x 2.89 mm with volume 23.87 µL and 2.86mm x 2.67 mm with volume 16.25 µL with T fuscobalteata The worker brood food volume of T iridipennis and T fuscobalteata gain 52.43 percent and 62.11 percent of brood cell capacity, respectively - The worker cell building duration of T iridipennis was 10.33 hours and T fuscobalteata was 7.13 hours in single cell and the type of cell building is Successive (Sc) - The worker food-discharged duration of both species are substantially distinct It means of T iridipennis Smith was 90.77 seconds at single cell, with the number of workers who inserted is 29.87 And T iridipennis Cameron was 94.07 seconds at single cell, with the number of workers who inserted is 24.03 The provisioning type was Semi-synchronous (Dm) - The egg-laying duration of T iridipennis and T fuscobalteata queens were 2.16 seconds per one cell, with egg dimension 0.92 mm x 0.37 mm and 50 1.53 seconds per one cell, with egg dimension 0.57 mm x 0.24 mm, in that order - The capping duration of T iridipennis and T fuscobalteata in single cell were significantly different It means 23.86 minutes and 5.15 minutes, with the total of operculators are to workers and to worker, respectively - The total number of cells that were provided eggs often lower that number of cells which were built completely 5.2 Recommendation - Stingless bees should be studied more in biological characteristics in order to enhancing pollinated affection and diversity maintain - Continue to conduct the research about the biological characteristics of T iridipennis, T fuscobalteata and other species to apply them pollination in greenhouse 51 REFERENCES Chinh T X and Sommeifer M J (2005) Production of sexuals in the stingless bees Trigona (Lepidotrigona) ventralis flavibasis Cockerll (Apidae, Meliponini) in northern Vietnam Apidologie 36 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T.fuscobalteata Cell width Count 30 30 ANOVA Source of Variation Between Groups Within Groups SS 41.20130667 1.491253333 Total 42.69256 Sum Average Variance 86.6 2.8867 0.0433 36.88 1.2293 0.0082 df 58 MS 41.201 0.0257 F 1602.5 P-value F crit 6.00608E-44 4.0069 59 Anova: Single Factor SUMMARY Groups V-Brood space (%) T.iridipennis T.fuscobalteata Count 30 30 ANOVA Source of Variation Between Groups Within Groups SS 1406.33 1806.13 Total 3212.46 Sum Average Variance 1863.25 62.1085 21.5917 1572.77 52.4257 40.6885 df 58 MS 1406.33 31.1401 F 45.1615 P-value 8.63508E-09 F crit 4.00687 59 56 Anova: Single Factor Anova: Single Factor V-brood food SUMMARY Groups Count Sum Average Variance T.iridipennis 30 376.6 12.55333333 4.199816092 T.fuscobalteata30 302.3 10.07666667 1.90391954 SUMMARY Groups ANOVA Source of VariationSS df Between Groups 92.00817 Within Groups 177.0083 58 Total 269.0165 MS F 92.00816667 30.14814933 3.051867816 T.iridipennis T.fuscobalteata V-total Count Sum Average Variance 30 716 23.86667 3.498851 30 487.5 16.25 4.357759 ANOVA P-value F crit Source of Variation SS 9.25924E-07 4.006873 Between Groups 870.2042 Within Groups 227.8417 59 Total df 1098.046 MS F P-value F crit 870.2042 221.5216 1.82115E-21 4.006873 58 3.928305 59 Anova: Single Factor SUMMARY Time of cell building Groups Count Sum Average Variance Hours 30 214 7.13333 3.63678 0 Hours 30 310 10.3333 11.2644 ANOVA Source of VariationSS Between Groups 153.6 Within Groups 432.133 Total Time 8h 9h 10h 11h 12h 13h 14h 585.733 df MS F P-value F crit 76.8 10.1302 0.00017 3.15884 57 7.58129 59 No cells are building at given time Tetragonula Tetragonula Iridipennis Fuscobalteata 11 10 5 18 17 17 17 15 15 15 57 15h 16h 17h 18h 19h 20h 21h 22h 23h 24h 1h 2h 3h 4h 5h 6h 7h 8h 3 4 4 4 6 11 12 15 15 17 16 17 T iridipennis 15 15 16 12 12 14 16 19 19 25 26 27 28 28 26 25 25 25 T fuscobalteata Date hour4h - Oct 20th 4h - Oct 27th 4h - Nov 6th 4h - Oct 144h th- Oct 21st 4h - Oct 29 th Started 6 1/4 built 7 1/2 built 6 Stages 3/4 built 2 8 4/4 built completed 13 13 12 40 32 33 total 10 15 18 28 28 30 58 Anova: Single Factor SUMMARY Groups Iridipennis Fulcobalteata EGG-Laying time Count 30 30 Sum ANOVA Source of Variation SS Between Groups 5.88440167 Within Groups 6.63556333 Total Average Variance 64.79 2.15966667 0.18669 46 1.53333333 0.04212 df MS F P-value F crit 5.88440167 51.4343 1.50816E-09 4.00687289 58 0.11440626 12.519965 Anova: Single Factor SUMMARY Groups 1st 2nd 3rd 4th 5th 6th 7th 8th 9th 10th 59 T.iridipennis Count 30 30 30 30 30 30 30 30 30 30 ANOVA Source of Variatio n Between Groups Within Groups SS 7116.795753 1711.802913 Total 8828.598667 Sum 467.87 357.13 218.03 163.43 127.3 72.72 32.7 30.64 27.67 26.11 df Average 15.59566667 11.90433333 7.267666667 5.447666667 4.243333333 2.424 1.09 1.021333333 0.922333333 0.870333333 MS 790.7550837 290 5.902768667 Variance 16.33389437 15.23681161 12.73512885 4.432335747 5.539898851 3.576397241 0.380317241 0.366225747 0.202025402 0.224651609 F P-value F crit 133.963421 7.83224E-98 1.912235867 299 59 Anova: Single Factor T.fuscobalteata SUMMARY Groups 1st 2nd 3rd 4th 5th 6th 7th 8th 9th 10th Count 30 30 30 30 30 30 30 30 30 30 ANOVA Source of Variation Between Groups Within Groups SS 198.2083 12.67221 Total 210.8805 Sum 101.89 102.06 87.84 88.05 57.27 56.96 41.86 43.78 40.08 43.87 df Average 3.396333 3.402 2.928 2.935 1.909 1.898667 1.395333 1.459333 1.336 1.462333 Variance 0.018003 0.029465 0.064754 0.066612 0.063223 0.043543 0.016715 0.021737 0.079763 0.033156 MS F P-value F crit 22.02315 503.9935 2.3E-171 1.912236 290 0.043697 299 60 Anova: Single Factor SUMMARY Groups T.iridipennis T.fuscobalteata Operculation duration in single cell (minutes) Count Sum 30 30 ANOVA Source of Variation Between Groups Within Groups SS 5253.768375 567.5936389 Total 5821.362014 Average Variance 715.85 154.4 23.86166667 19.33742 5.146666667 0.234778 df MS F P-value 5253.768375 536.8604 5.26E-31 9.786097222 58 F crit 4.006872886 59 Anova: Single Factor SUMMARY Groups Number of workers operculated Count T.iridipennis T.fuscobalteata ANOVA Source of Variation Between Groups Within Groups Total 30 Sum Average Variance 58 1.933333 1.236782 30 39 SS df 1.3 0.217241 6.016666667 42.16666667 MS F P-value F crit 6.016667 8.275889 0.005613 4.006873 58 0.727011 48.18333333 59 61 Anova: Single Factor SUMMARY Groups Egg length(mm) Count T.iridipennis T.fuscobalteata ANOVA Source of Variation Between Groups Within Groups Total 31 31 SS 1.920512 0.099934142 2.020446142 Sum 28.6336 17.7216 df 60 Average Variance 0.923664516 0.001666 0.571664516 0.001666 MS F 1.920512 1153.067 0.001665569 P-value 7.07464E-41 F crit 4.001191377 61 62 Anova: Single Factor SUMMARY Groups Count T.iridipennis T.fuscobalteata ANOVA Source of Variation Between Groups Within Groups Brood-food discharged duration (seconds) Sum Average Variance 30 2723 90.76667 1601.150575 30 2821 94.03333 382.8609195 SS 160.0666667 57536.33333 Total df 57696.4 MS 160.0667 58 992.0057 F 0.161356592 P-value F crit 0.689386932 4.006873 59 Anova: Single Factor SUMMARY Egg width(mm) Groups Count T.iridipennis T.fuscobalteata ANOVA Source of Variation Between Groups Within Groups Total 31 Sum 11.4272 Average Variance 0.368619355 0.000398 31 7.4592 0.240619355 0.000398 df MS F 0.253952 637.6203 0.000398281 SS 0.253952 0.023896857 0.277848857 60 P-value 1.16384E-33 F crit 4.001191377 61 Anova: Single Factor SUMMARY Groups T.iridipennis T.fuscobalteata ANOVA Source of Variation Between Groups Within Groups Total No workers who inserted Count Sum Average Variance 30 896 29.86667 53.42989 30 721 24.03333 48.72299 SS 510.4167 2962.433 3472.85 df MS F P-value F crit 510.4167 9.993193 0.002498 4.006873 58 51.07644 59 63 64