Matchmakers' Tricky Task: A Comparative Review of Instrumental Insemination in Apis mellifera and the Associated Challenges in Apis cerana indica
DOI:
https://doi.org/10.13102/sociobiology.v73i3.12663Keywords:
honey bee breeding, drone congregation areas (DCA), reproductive behavior, controlled mating, quenn mating biologyAbstract
Instrumental insemination is an important tool in honey bee breeding as it provides controlled mating, genetic improvement, and conservation of valuable traits. This technique is widely practised in the European honey bee (Apis mellifera), but its application in the Indian honey bee (Apis cerana indica) is limited due to species-specific biological, behavioural and technical challenges. In this review, honey bee biology relevant to instrumental insemination is analysed, and a comparative assessment of the technique in A. mellifera and A. cerana indica is presented. Key aspects such as caste differentiation and sex determination, drone congregation areas, natural mating behaviour, procedures of instrumental insemination, and factors influencing insemination success such as queen age, semen collection and handling, carbon dioxide exposure, and post-insemination management are discussed. In A. cerana indica, smaller body size, reduced spermathecal capacity, and increased sensitivity to handling are identified as major constraints affecting insemination efficiency and post-insemination performance. Methodological limitations, procedural standardization, and operator-dependent variability influencing insemination outcomes are also analyzed. Understanding these interspecific differences is essential for refining species-specific protocols and expanding the application of instrumental insemination for selective breeding, genetic conservation, and sustainable apiculture. The review also outlines future research prospects aimed at improving efficiency and promoting wider adoption of instrumental insemination, particularly in indigenous honey bee species.
Downloads
References
Baer, B., Collins, J., Maalaps, K. & den Boer, S.P. (2016). Sperm use economy of honeybee (Apis mellifera) queens. Ecology and Evolution, 6: 2877-2885. DOI: https://doi.org/10.1002/ece3.2075
Beye, M., Hasselmann, M., Fondrk, M.K., Page, R.E. & Omholt, S.W. (2003). The gene csd is the primary signal for sexual development in the honeybee and encodes an SR-type protein. Cell, 114: 419-429. DOI: https://doi.org/10.1016/S0092-8674(03)00606-8
Bieńkowska, M., Węgrzynowicz, P., Panasiuk, B., Gerula, D. & Loc, K. (2008). Influence of the age of honey bee queens and dose of semen on condition of instrumentally inseminated queens kept in cages with 25 worker bees in bee colonies. Journal of Apicultural Science, 52: 23-34.
Bolten, A.B. & Harbo, J.R. (1982). Numbers of spermatozoa in the spermatheca of the queen honeybee after multiple inseminations with small volumes of semen. Journal of Apicultural Research, 21: 7-10. DOI: https://doi.org/10.1080/00218839.1982.11100509
Borah, P., Rahman, A. & Deka, M. (2024). Performance of artificially and naturally inseminated honey bee queens (Apis cerana indica). Journal of Advances in Biology & Biotechnology, 27: 552-562. DOI: https://doi.org/10.9734/jabb/2024/v27i6915
Brandstaetter, A.S., Bastin, F. & Sandoz, J.-C. (2014). Honeybee drones are attracted by groups of consexuals in a walking simulator. Journal of Experimental Biology, 217: 1278-1285. DOI: https://doi.org/10.1242/jeb.094292
Brutscher, L.M., Baer, B. & Niño, E.L. (2019). Putative drone copulation factors regulating honey bee (Apis mellifera) queen reproduction and health: A review. Insects, 10: 8. DOI: https://doi.org/10.3390/insects10010008
Büchler, R., Andonov, S., Bienefeld, K., Costa, C., Hatjina, F., Kezic, N., Kryger, P., Spivak, M., Uzunov, A. & Wilde, J. (2013). Standard methods for rearing and selection of Apis mellifera queens. Journal of Apicultural Research, 52: 1-30. DOI: https://doi.org/10.3896/IBRA.1.52.1.07
Buescu, E., Gurău, M.R. & Bîrțoıu, A. (2015). Artificial insemination on Apis mellifera – aspects of artificially inseminated queen performance and factors that may affect their performance. Proceedings of the Romanian Academy, Series B, Supplement 1: 21-24.
Burley, L.M., Fell, R.D. & Saacke, R.G. (2008). Survival of honey bee (Hymenoptera: Apidae) spermatozoa incubated at room temperature from drones exposed to miticides. Journal of Economic Entomology, 101: 1081-1087. DOI: https://doi.org/10.1093/jee/101.4.1081
Chuda-Mickiewicz, B., Czekońska, K., Samborski, J. & Rostecki, P. (2012). Success rates for instrumental insemination of carbon dioxide and nitrogen anaesthetised honey bee (Apis mellifera) queens. Journal of Apicultural Research, 51: 74-77. DOI: https://doi.org/10.3896/IBRA.1.51.1.09
Cobey, S.W. (2007). Comparison studies of instrumentally inseminated and naturally mated honey bee queens and factors affecting their performance. Apidologie, 38: 390-410. DOI: https://doi.org/10.1051/apido:2007029
Cobey, S.W. (2016). An introduction to instrumental insemination of honey bee queens. Bee World, 93: 33-36. DOI: https://doi.org/10.1080/0005772X.2016.1222790
Cobey, S.W., Tarpy, D.R. & Woyke, J. (2013). Standard methods for instrumental insemination of Apis mellifera queens. Journal of Apicultural Research, 52: 1-18. DOI: https://doi.org/10.3896/IBRA.1.52.4.09
Collins, A.M. (2000). Relationship between semen quality and performance of instrumentally inseminated honey bee queens. Apidologie, 31: 421-429. DOI: https://doi.org/10.1051/apido:2000132
Czekońska, K. (2009). The effect of different concentrations of carbon dioxide (CO₂) in a mixture with air or nitrogen upon the survival of the honey bee (Apis mellifera). Journal of Apicultural Research, 48: 67-71. DOI: https://doi.org/10.3896/IBRA.1.48.1.13
da Silva Morais, L., de Araujo Neto, E.R., da Silva, A.M., Bezerra, L.G.P., da Cunha, A.F.S., de Sousa Chagas, N.O., dos Santos, R.P., Bergamo, G.C., Façanha, D.A.E., Gramacho, K.P. & Silva, A.R. (2023). Africanized honeybee (Apis mellifera) semen freezing using Tris-based and Collins extenders. Tropical Animal Health and Production, 55: 329. DOI: https://doi.org/10.1007/s11250-023-03762-6
Ding, G., Xu, H., Oldroyd, B.P. & Gloag, R.S. (2017). Extreme polyandry aids the establishment of invasive populations of a social insect. Heredity, 119: 381-387. DOI: https://doi.org/10.1038/hdy.2017.49
Dziechciarz, P., Domaciuk, M., Pyz-Łukasik, R., Olszewski, K. & Borsuk, G. (2025). Breakthrough research on reinsemination of bee queens with imaging of reproductive system elements. Scientific Reports, 15: 20810. DOI: https://doi.org/10.1038/s41598-025-03278-z
Evans, J.D. & Wheeler, D.E. (1999). Differential gene expression between developing queens and workers in the honey bee, Apis mellifera. Proceedings of the National Academy of Sciences, 96: 5575-5580. DOI: https://doi.org/10.1073/pnas.96.10.5575
Fischer, F. (1990). External influences on the filling of the spermatheca with sperm. Apidologie, 21: 359-360.
Food and Agriculture Organization (FAO). (2018). The importance of bees and other pollinators for food and agriculture. Rome: FAO.
Free, J.B. & Williams, I.H. (1975). Factors determining the rearing and rejection of drones by the honeybee colony. Animal Behaviour, 23: 650-675. DOI: https://doi.org/10.1016/0003-3472(75)90143-8
Gąbka, J. & Cobey, S.W. (2018). Factors, based on common practices, affecting the results of instrumental insemination of honey bee queens. Apidologie, 49: 773-780. DOI: https://doi.org/10.1007/s13592-018-0606-y
Gąbka, J., Gąbka, J. & Zajdel, B. (2022). Effect of time between carbon dioxide treatments on the onset of oviposition in queen honey bees. Journal of Apicultural Science, 66: 171-178. DOI: https://doi.org/10.2478/jas-2022-0011
Gąbka, J., Gąbka, J. & Zajdel, B. (2025). Re-insemination of egg laying queen honey bees. Journal of Apicultural Science, 69: 59-61. DOI: https://doi.org/10.2478/jas-2025-0002
Goins, A. & Schneider, S. (2013). Drone “quality” and caste interactions in the honey bee, Apis mellifera L. Insectes Sociaux, 60: 453-461. DOI: https://doi.org/10.1007/s00040-013-0310-x
Gotoh, A. & Sasaki, K. (2021). Caste differentiation of spermatheca and organs related to sperm use and oviposition in the honeybee, Apis mellifera. Apidologie, 52: 262-271. DOI: https://doi.org/10.1007/s13592-020-00815-9
Gries, M. & Koeniger, N. (1996). Straight forward to the queen: pursuing honeybee drones (Apis mellifera L.) adjust their body axis to the direction of the queen. Journal of Comparative Physiology A, 179: 539-544. DOI: https://doi.org/10.1007/BF00192319
Grossman, E. (2013). Declining bee populations pose a threat to global agriculture. Yale Environment 360. https://e360.yale.edu/features/declining_bee_populations_pose_a_threat_to_global_agriculture (accessed date: 14 January 2026).
Güler, A., Önder, H., Kavak, G., Toktay, G., Uğurlutepe, E., Biyik, S. & Aydin, A. (2022). The effects of instrumental insemination on selected and unselected breeding characteristics in honeybee (Apis mellifera L.). Apidologie, 53: 35. DOI: https://doi.org/10.1007/s13592-022-00947-0
Halvacı, E., Kozak, T., Gül, M., Kars, H. & Şen, F. (2023). Bee anatomy: a comprehensive overview of bee morphology and physiology. Journal of Scientific Reports-B, 8: 1-19.
Hepburn, H.R. & Radloff, S.E. (2011). Honeybees of Asia. Springer Science & Business Media. DOI: https://doi.org/10.1007/978-3-642-16422-4
Hopkins, B.K. & Herr, C. (2010). Factors affecting the successful cryopreservation of honey bee (Apis mellifera) spermatozoa. Apidologie, 41: 548-556. DOI: https://doi.org/10.1051/apido/20010006
Jackson, J.T., Tarpy, D.R. & Fahrbach, S.E. (2011). Histological estimates of ovariole number in honey bee queens, Apis mellifera, reveal lack of correlation with other queen quality measures. Journal of Insect Science, 11: 82. DOI: https://doi.org/10.1673/031.011.8201
Jones, J.C., Du, Z.G., Bernstein, R., Meyer, M., Hoppe, A., Schilling, E., Ableitner, M., Juling, K., Dick, R., Strauss, A.S. & Bienefeld, K. (2020). Tool for genomic selection and breeding to evolutionary adaptation: development of a 100K single nucleotide polymorphism array for the honey bee. Ecology and Evolution, 10: 6246-6256. DOI: https://doi.org/10.1002/ece3.6357
Kahya, Y. (2020). HRM analysis of spermathecal contents to determine the origin of drones that inseminated honey bee queens. Journal of Apicultural Science, 64: 241-249. DOI: https://doi.org/10.2478/jas-2020-0018
Kahya, Y. & Gençer, H.V. (2022). Temporal variation in the viability of spermatozoa in the spermathecae of queen honey bees (Apis mellifera L.). Turkish Journal of Agriculture-Food Science and Technology, 10: 319-322. DOI: https://doi.org/10.24925/turjaf.v10i2.319-322.4894
Kama, O. & Shpigler, H.Y. (2025). Social and nutritional factors controlling the growth of honey bee (Apis mellifera) queens. PLoS ONE, 20: e0310608. DOI: https://doi.org/10.1371/journal.pone.0310608
Kapil, R. (1962). Anatomy and histology of the male reproductive system of Apis indica Fab. (Apidae, Hymenoptera). Insectes Sociaux, 9: 73-90. DOI: https://doi.org/10.1007/BF02224581
Kaya, A., Uysal, O. & Akyol, N. (2025). Improving drone sperm cryopreservation: investigating cryoprotectant combinations and PVP supplementation. Cryobiology, 121: 105318. DOI: https://doi.org/10.1016/j.cryobiol.2025.105318
Khalifa, S.A.M., Elshafiey, E.H., Shetaia, A.A., Abd El-Wahed, A.A., Algethami, A.F., Musharraf, S.G., AlAjmi, M.F., Zhao, C., Masry, S.H.D., Abdel-Daim, M.M., Halabi, M.F., Kai, G., Al Naggar, Y., Bishr, M., Diab, M.A.M. & El-Seedi, H.R. (2021). Overview of bee pollination and its economic value for crop production. Insects, 12: 688. DOI: https://doi.org/10.3390/insects12080688
Khan, K.A., Rafique, M.K., Lashari, M.A., Iqbal, A., Mahmood, R., Ahmed, A.M., Khoso, F.N., Ahmad, S., AL-Shehri, B.M., Mohammed, M.E.A. & Ghramh, H.A. (2022). Instrumental insemination: A nontraditional technique to produce superior quality honey bee (Apis mellifera) queens. Journal of King Saud University -Science, 34: 102077. DOI: https://doi.org/10.1016/j.jksus.2022.102077
Klein, C.D., Kozii, I.V., Wood, S.C., Koziy, R.V., Zabrodski, M.W., Dvylyuk, I., de Mattos, I.M., Moshynskyy, I., Honaramooz, A. & Simko, E. (2021). Testicular changes of honey bee drones, Apis mellifera (Hymenoptera: Apidae), during sexual maturation. Journal of Insect Science, 21: 1-12. DOI: https://doi.org/10.1093/jisesa/ieab049
Kocher, S.D. & Grozinger, C.M. (2011). Cooperation, conflict, and the evolution of queen pheromones. Journal of Chemical Ecology, 37: 1263-1275. DOI: https://doi.org/10.1007/s10886-011-0036-z
Koeniger, G. (1988). Mating flights of honey bee drones (Apis mellifera L.). A film documentation. In W. Nachtigall (Ed.), The flying honeybee (pp. 29-34). Stuttgart: Gustav Fischer Verlag.
Koeniger, G. (1990). The role of the mating sign in honey bees, Apis mellifera L.: does it hinder or promote multiple mating? Animal Behaviour, 39: 444-449. DOI: https://doi.org/10.1016/S0003-3472(05)80407-5
Koeniger, N. & Koeniger, G. (2000). Reproductive isolation among species of the genus Apis. Apidologie, 31: 313-339. DOI: https://doi.org/10.1051/apido:2000125
Koeniger, N., Koeniger, G., Gries, M. & Tingek, S. (2005a). Drone competition at drone congregation areas in four Apis species. Apidologie, 36: 211-221. DOI: https://doi.org/10.1051/apido:2005011
Koeniger, N., Koeniger, G. & Pechhacker, H. (2005b). The nearer the better? Drones (Apis mellifera) prefer nearer drone congregation areas. Insectes Sociaux, 52: 31-35. DOI: https://doi.org/10.1007/s00040-004-0763-z
Koeniger, N. & Wijayagunasekera, H. (1976). Time of drone flight in the three Asiatic honeybee species (Apis cerana, Apis florea, Apis dorsata). Journal of Apicultural Research, 15: 67-71. DOI: https://doi.org/10.1080/00218839.1976.11099837
Kozii, I.V., Wood, S.C., Koziy, R.V. & Simko, E. (2022). Histomorphological description of the reproductive system in mated honey bee queens. Journal of Apicultural Research, 61: 114-126. DOI: https://doi.org/10.1080/00218839.2021.1900636
Kraus, F., Neumann, P., Van Praagh, J. & Moritz, R. (2004). Sperm limitation and the evolution of extreme polyandry in honeybees (Apis mellifera L.). Behavioral Ecology and Sociobiology, 55: 494-501. DOI: https://doi.org/10.1007/s00265-003-0706-0
Lago, D.C., Martins, J.R., Dallacqua, R.P., Santos, D.E., Bitondi, M.M. & Hartfelder, K. (2020). Testis development and spermatogenesis in drones of the honey bee, Apis mellifera L. Apidologie, 51: 935-955. DOI: https://doi.org/10.1007/s13592-020-00773-2
Lago, D.C., Nora, L.C., Hasselmann, M. & Hartfelder, K. (2023). Positive selection in cytochrome P450 genes is associated with gonad phenotype and mating strategy in social bees. Scientific Reports, 13: 5921. DOI: https://doi.org/10.1038/s41598-023-32898-6
Le Conte, Y. & Hefetz, A. (2008). Primer pheromones in social Hymenoptera. Annual Review of Entomology, 53: 523-542. DOI: https://doi.org/10.1146/annurev.ento.52.110405.091434
Lin, Z., Yang, L., Wang, Z., Wang, K., Niu, Q. & Ji, T. (2025). Honey bee breeding and breed: advancements, challenges, and prospects. Animal Research and One Health, 3: 350-357. DOI: https://doi.org/10.1002/aro2.70003
Liu, Z., Liu, F., Li, G., Chi, X., Wang, Y., Wang, H., Ma, L., Han, K., Zhao, G., Guo, X. & Xu, B. (2020). Metabolite support of long-term storage of sperm in the spermatheca of honeybee (Apis mellifera) queens. Frontiers in Physiology, 11: 574856. DOI: https://doi.org/10.3389/fphys.2020.574856
Lodesani, M., Balduzzi, D. & Galli, A. (2004). Functional characterisation of semen in honeybee queen (A. m. ligustica S.) spermatheca and efficiency of the diluted semen technique in instrumental insemination. Italian Journal of Animal Science, 3: 385-392. DOI: https://doi.org/10.4081/ijas.2004.385
Loper, G.M. & Taylor Jr, O.R. (1987). Detection and monitoring of honeybee drone congregation areas by radar. Apidologie, 18: 163-172. DOI: https://doi.org/10.1051/apido:19870206
Mackensen, O. & Tucker, K.W. (1970). Instrumental insemination of queen bees. Agriculture Handbook No. 390. Washington, DC: US Agricultural Research Service.
Mansour, M.A., Mazeed, A.M., Abd-Alfattah, M.A. & Elenany, Y.E. (2025). Effect of introducing methods of honey bee Apis mellifera L. queens on the number of spermatozoa after instrumental insemination. Indian Journal of Entomology, 88: 493-496. DOI: https://doi.org/10.55446/IJE.2025.2878
Maucourt, S., Rousseau, A., Fortin, F., Robert, C. & Giovenazzo, P. (2023). Observation of genetic gain with instrumental insemination of honeybee queens. Insects, 14: 301. DOI: https://doi.org/10.3390/insects14030301
McAfee, A., Degueldre, F., Hoover, S.E., Aron, S. & Foster, L.J. (2025). Mating-induced patterns of spermathecal fluid protein expression in two eusocial insect species, Lasius niger and Apis mellifera. Scientific Reports, 15: 38917. DOI: https://doi.org/10.1038/s41598-025-22689-6
Meixner, M.D., Costa, C., Kryger, P., Hatjina, F., Bouga, M., Ivanova, E. & Büchler, R. (2010). Conserving diversity and vitality for honey bee breeding. Journal of Apicultural Research, 49: 85-92. DOI: https://doi.org/10.3896/IBRA.1.49.1.12
Metz, B.N. & Tarpy, D.R. (2019). Reproductive senescence in drones of the honey bee (Apis mellifera). Insects, 10: 11. DOI: https://doi.org/10.3390/insects10010011
Metz, B.N. & Tarpy, D.R. (2021). Reproductive and morphological quality of commercial honey bee (Hymenoptera: Apidae) drones in the United States. Journal of Insect Science, 21: 2. DOI: https://doi.org/10.1093/jisesa/ieab048
Michener, C.D. (1974). The social behavior of the bees: a comparative study. Cambridge, Massachusetts: Harvard University Press.
Moškrič, A., Pavlin, A., Mole, K., Marinč, A., Bubnič, J., Opara, A., Kovačić, M., Puškadija, Z., Uzunov, A., Andonov, S., Dahle, B. & Prešern, J. (2023). Cutting corners: the impact of storage and DNA extraction on quality and quantity of DNA in honeybee (Apis mellifera) spermatheca. Frontiers in Physiology, 14: 1139269. DOI: https://doi.org/10.3389/fphys.2023.1139269
Nelson, D. (1989). Assessment of queen quality in honey bee queens. Canadian Beekeeper, 14: 207-208.
Nur, Z., Seven Çakmak, S., Çakmak, İ., Onder, N.T., Gokçe, E., Ustuner, B., Alcay, S., Toker, M.B. & Soylu, M.K. (2020). Effects of trehalose supplementation on post-thaw sperm quality of honey bee drones. Online Journal of Animal and Feed Research, 10: 191-196. DOI: https://doi.org/10.51227/ojafr.2020.27
Oldroyd, B.P. & Nanork, P. (2009). Conservation of Asian honey bees. Apidologie, 40: 296-312. DOI: https://doi.org/10.1051/apido/2009021
Özkök, A.O. & Selçuk, M. (2020). Sperm storage and artificial insemination in honey bees. International Journal of Science Letters, 2: 12-25. DOI: https://doi.org/10.38058/ijsl.661629
Page Jr, R.E. (1981). Protandrous reproduction in honey bees. Environmental Entomology, 10: 359-362. DOI: https://doi.org/10.1093/ee/10.3.359
Palmer, K.A. & Oldroyd, B.P. (2000). Evolution of multiple mating in the genus Apis. Apidologie, 31: 235-248. DOI: https://doi.org/10.1051/apido:2000119
Pan, L., Wang, Z., Zhong, S., Xu, T., Chen, W., Cheng, F. & Zeng, Z. (2025). Agxt2l-mediated glycerophospholipid metabolism in trophocytes explains Apis mellifera queen’s higher oviposition over A. cerana. Communications Biology, 8: 1091. DOI: https://doi.org/10.1038/s42003-025-08526-6
Pasho, D.J., Applegate, J.R. & Hopkins, D.I. (2021). Diseases and pests of honey bees (Apis mellifera). Veterinary Clinics: Food Animal Practice, 37: 401-412. DOI: https://doi.org/10.1016/j.cvfa.2021.06.001
Pettis, J.S., Rice, N., Joselow, K., vanEngelsdorp, D. & Chaimanee, V. (2016). Colony failure linked to low sperm viability in honey bee (Apis mellifera) queens and an exploration of potential causative factors. PLoS ONE, 11: e0147220. DOI: https://doi.org/10.1371/journal.pone.0147220
Phiancharoen, M., Wongsiri, S., Koeniger, N. & Koeniger, G. (2004). Instrumental insemination of Apis mellifera queens with hetero- and conspecific spermatozoa results in different sperm survival. Apidologie, 35: 503-511. DOI: https://doi.org/10.1051/apido:2004043
Plate, M., Bernstein, R., Hoppe, A. & Bienefeld, K. (2019). The importance of controlled mating in honeybee breeding. Genetics Selection Evolution, 51: 74. DOI: https://doi.org/10.1186/s12711-019-0518-y
Power, K., Martano, M., Altamura, G. & Maiolino, P. (2020). Histopathological findings in testes from apparently healthy drones of Apis mellifera ligustica. Veterinary Sciences, 7: 124. DOI: https://doi.org/10.3390/vetsci7030124
Punchihewa, R., Koeniger, N. & Koeniger, G. (1990). Congregation of Apis cerana indica Fabricius 1798 drones in the canopy of trees in Sri Lanka. Apidologie, 21: 201-208. DOI: https://doi.org/10.1051/apido:19900305
Rajamohan, A., Danka, R.G., Hopkins, B.K. & Rinehart, J.P. (2020). A non-activating diluent to prolong in vitro viability of Apis mellifera spermatozoa: effects on cryopreservation and on egg fertilization. Cryobiology, 92: 124-129. DOI: https://doi.org/10.1016/j.cryobiol.2019.11.045
Rangel, J. & Fisher, A. (2019). Factors affecting the reproductive health of honey bee (Apis mellifera) drones – A review. Apidologie, 50: 759-778. DOI: https://doi.org/10.1007/s13592-019-00684-x
Rangel, J., Shepherd, T.F., Gonzalez, A.N., Hillhouse, A., Konganti, K. & Ing, N.H. (2021). Transcriptomic analysis of the honey bee (Apis mellifera) queen spermathecae reveals genes that may be involved in sperm storage after mating. PLoS ONE, 16: e0244648. DOI: https://doi.org/10.1371/journal.pone.0244648
Rhodes, J.W., Harden, S., Spooner-Hart, R., Anderson, D.L. & Wheen, G. (2011). Effects of age, season and genetics on semen and sperm production in Apis mellifera drones. Apidologie, 42: 29-38. DOI: https://doi.org/10.1051/apido/2010026
Richard, F.-J., Tarpy, D.R. & Grozinger, C.M. (2007). Effects of insemination quantity on honey bee queen physiology. PLoS ONE, 2: e980. DOI: https://doi.org/10.1371/journal.pone.0000980
Roberts, W.C. (1944). Multiple mating of queen bees proved by progeny and flight tests. Gleanings in Bee Culture, 72: 225-259.
Roberts, K., Evison, S., Baer, B. & Hughes, W. (2015). The cost of promiscuity: sexual transmission of Nosema microsporidian parasites in polyandrous honey bees. Scientific Reports, 5: 10982. DOI: https://doi.org/10.1038/srep10982
Rothenbuhler, W.C. (1958). Genetics and breeding of the honey bee. Annual Review of Entomology, 3: 161-180. DOI: https://doi.org/10.1146/annurev.en.03.010158.001113
Rousseau, A., Fournier, V. & Giovenazzo, P. (2015). Apis mellifera (Hymenoptera: Apidae) drone sperm quality in relation to age, genetic line, and time of breeding. The Canadian Entomologist, 147: 702-711. DOI: https://doi.org/10.4039/tce.2015.12
Ruttner, F., Woyke, J. & Koeniger, N. (1972). Reproduction in Apis cerana 1. Mating behaviour. Journal of Apicultural Research, 11: 141-146. DOI: https://doi.org/10.1080/00218839.1972.11099714
Ruttner, F., Woyke, J. & Koeniger, N. (1973). Reproduction in Apis cerana 2. Reproductive organs and natural insemination. Journal of Apicultural Research, 12: 21-34. DOI: https://doi.org/10.1080/00218839.1973.11099727
Sawarkar, A.B. & Tembhare, D.B. (2010). Synthesis and chemical composition of mucus gland secretions in Apis cerana indica. Apidologie, 41: 488-496. DOI: https://doi.org/10.1051/apido/2009078
Sawarkar, A.B. & Tembhare, D.B. (2014). Development and secretory nature of seminal vesicle during sexual maturation in Indian honeybee, Apis cerana indica F. (Hymenoptera: Apidae). Journal of Entomology and Zoology Studies, 2: 105-109.
Sawarkar, A.B. & Tembhare, D.B. (2015). Testis morphology and spermatogenesis in the Indian honeybee, Apis cerana indica F. (Hymenoptera: Apidae). Journal of Entomology and Zoology Studies, 3: 489-492.
Schlüns, H., Moritz, R.F., Neumann, P., Kryger, P. & Koeniger, G. (2005). Multiple nuptial flights, sperm transfer and the evolution of extreme polyandry in honeybee queens. Animal Behaviour, 70: 125-131. DOI: https://doi.org/10.1016/j.anbehav.2004.11.005
Shweta, K., Saminathan, V.R., Sowmiya, C., Preetha, G., Srinivasan, M.R., Baskaran, V. & Manivannan, N. (2025). Flying sperm: the indispensable component of the instrumental insemination of honey bees: a review. Indian Journal of Animal Research, 59: 1-10. DOI: https://doi.org/10.18805/IJAR.B-5384
Skowronek, W., Kruk, C. & Klopot, J. (2002). Factors affecting oviposition of artificially inseminated honeybee queens. Journal of Apicultural Science, 46: 85-95.
Slater, G.P., Smith, N.M. & Harpur, B.A. (2021). Prospects in connecting genetic variation to variation in fertility in male bees. Genes, 12: 1251. DOI: https://doi.org/10.3390/genes12081251
Smilga-Spalvina, A., Spalvins, K. & Veidenbergs, I. (2023). Review of sustainable cryopreservation and above-freezing storage solutions of European honey bee Apis mellifera drone semen. Rigas Tehniskas Universitates Zinatniskie Raksti, 27: 177-194. DOI: https://doi.org/10.2478/rtuect-2023-0014
Stoian, R.O., Mălinaş, C., Botha, M. & Petrescu-Mag, I.V. (2018). Technical, sanitary and environmental sequences to improve artificial insemination of honey bee, Apis mellifera. Part I. Experimental results. Animal Biology & Animal Husbandry, 10: 122.
Taylor, M.A., Guzmán-Novoa, E., Morfin, N. & Buhr, M.M. (2009). Improving viability of cryopreserved honey bee (Apis mellifera L.) sperm with selected diluents, cryoprotectants, and semen dilution ratios. Theriogenology, 72: 149-159. DOI: https://doi.org/10.1016/j.theriogenology.2009.02.012
Tirfie, A.M. & Getachew, A. (2024). Role of honeybees to crop pollination in Ethiopia: a review. Agricultural Reviews, 45: 142-145. DOI: https://doi.org/10.18805/ag.RF-283
Tsvetkov, N., Samson-Robert, O., Sood, K., Patel, H.S., Malena, D.A., Gajiwala, P.H., Maciukiewicz, P., Fournier, V. & Zayed, A. (2017). Chronic exposure to neonicotinoids reduces honey bee health near corn crops. Science, 356: 1395-1397. DOI: https://doi.org/10.1126/science.aam7470
Van der Sluijs, J.P. & Vaage, N.S. (2016). Pollinators and global food security: the need for holistic global stewardship. Food Ethics, 1: 75-91. DOI: https://doi.org/10.1007/s41055-016-0003-z
Verma, L.R. (1991). Beekeeping in integrated mountain development: economic and scientific perspectives. New Delhi: Oxford & IBH Pub. Co. DOI: https://doi.org/10.53055/ICIMOD.49
Vung, N.N., Kim, I., Lee, M.-Y., Kim, H.K., Kim, D.W. & Choi, Y.S. (2018). Impact of confinement and population size on the instrumentally inseminated queen’s performance of Apis cerana species in South Korea. Journal of Apiculture, 33: 251-260. DOI: https://doi.org/10.17519/apiculture.2018.11.33.4.251
Vung, N.N., Lee, M.L., Kim, H.K., Byeon, K.H. & Choi, Y.S. (2016). Efficiency of artificial insemination for breeding Apis cerana in Korea. Journal of Apiculture, 31: 323-330. DOI: https://doi.org/10.17519/apiculture.2016.11.31.4.323
Wakgari, M. & Yigezu, G. (2021). Honeybee keeping constraints and future prospects. Cogent Food & Agriculture, 7: 1872192. DOI: https://doi.org/10.1080/23311932.2021.1872192
Wegener, J. & Bienefeld, K. (2012). Toxicity of cryoprotectants to honey bee semen and queens. Theriogenology, 77: 600-607. DOI: https://doi.org/10.1016/j.theriogenology.2011.08.036
Wegener, J., May, T., Kamp, G. & Bienefeld, K. (2014). A successful new approach to honeybee semen cryopreservation. Cryobiology, 69: 236-242. DOI: https://doi.org/10.1016/j.cryobiol.2014.07.011
Winston, M.L. (1991). The biology of the honey bee. Cambridge, Massachusetts: Harvard University Press.
Woyke, J. (1962). Natural and artificial insemination of queen honeybees. Bee World, 43: 21-25. DOI: https://doi.org/10.1080/0005772X.1962.11096922
Woyke, J. (1971). Correlations between the age at which honeybee brood was grafted, characteristics of the resultant queens, and results of insemination. Journal of Apicultural Research, 10: 45-55. DOI: https://doi.org/10.1080/00218839.1971.11099669
Woyke, J. (1973). Instrumental insemination of Apis cerana indica queens. Journal of Apicultural Research, 12: 151-158. DOI: https://doi.org/10.1080/00218839.1973.11099743
Woyke, J. (1975). Natural and instrumental insemination of Apis cerana indica in India. Journal of Apicultural Research, 14: 153-159. DOI: https://doi.org/10.1080/00218839.1975.11099820
Woyke, J. (1983). Dynamics of entry of spermatozoa into the spermatheca of instrumentally inseminated queen honeybees. Journal of Apicultural Research, 22: 150-154. DOI: https://doi.org/10.1080/00218839.1983.11100579
Woyke, J. (2008). Why the eversion of the endophallus of honey bee drone stops at the partly everted stage and significance of this. Apidologie, 39: 627-636. DOI: https://doi.org/10.1051/apido:2008046
Woyke, J., Fliszkiewicz, C. & Jasiński, Z. (2001). Prevention of natural mating of instrumentally inseminated queen honeybees by proper method of instrumental insemination. Journal of Apicultural Science, 45: 101-114.
Woyke, J. & Jasiński, Z. (1976). The influence of age on the results of instrumental insemination of honeybee queens. Apidologie, 7: 301-306. DOI: https://doi.org/10.1051/apido:19760402
Woyke, J. & Jasiński, Z. (1990). Effect of the number of attendant worker bees on the initiation of egg laying by instrumentally inseminated queens kept in small nuclei. Journal of Apicultural Research, 29: 101-106. DOI: https://doi.org/10.1080/00218839.1990.11101204
Yadav, S., Kumar, Y. & Jat, B.L. (2017). Honeybee: diversity, castes and life cycle. In Omkar (Ed.), Industrial entomology (pp. 5-34). Singapore: Springer Singapore. DOI: https://doi.org/10.1007/978-981-10-3304-9_2
Yániz, J.L., Silvestre, M.A. & Santolaria, P. (2020). Sperm quality assessment in honey bee drones. Biology, 9: 174. DOI: https://doi.org/10.3390/biology9070174
Yıldız, B.İ. & Karabağ, K. (2025). Targeted gene editing in honey bees using liposome-based CRISPR-Cas9. Biochemical Genetics. 64: 4477-4492. DOI: https://doi.org/10.1007/s10528-025-11233-w
Yoshida, T., Saito, J. & Kajigaya, N. (1994). The mating flight times of native Apis cerana japonica Radoszkowski and introduced Apis mellifera L. in sympatric conditions. Apidologie, 25: 353-360. DOI: https://doi.org/10.1051/apido:19940401
Zapata‐Hernández, G., Gajardo‐Rojas, M., Calderón‐Seguel, M., Muñoz, A.A., Yáñez, K.P., Requier, F., Fontúrbel, F.E., Ormeño-Arriagada, P.I. & Arrieta, H. (2024). Advances and knowledge gaps on climate change impacts on honey bees and beekeeping: a systematic review. Global Change Biology, 30: e17219. DOI: https://doi.org/10.1111/gcb.17219
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2026 S. R. Adheena, M. R. Srinivasan, V. R. Saminathan, N. Manikanda Boopathi, S. Marimuthu, V. Deva Dharshini, M. Dharani, R .K. Aliya Mumthas, G. Mahesh Boopathy

This work is licensed under a Creative Commons Attribution 4.0 International License.
Sociobiology is a diamond open access journal which means that all content is freely available without charge to the user or his/her institution. Users are allowed to read, download, copy, distribute, print, search, or link to the full texts of the articles in this journal without asking prior permission from the publisher or the author. This is in accordance with the BOAI definition of open access.
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).

eISSN 2447-8067









