Temporal Variation in the Viability of Spermatozoa in the Spermathecae of Queen Honey Bees (Apis mellifera L.)

The storage of spermatozoa and keeping alive for years in the spermatheca by the honey bee queen is a phenomenon allowing her to fertilize eggs throughout her life. In this study, the queens that were 2-week, 1-year, and 2-year-old were analyzed to determine the viability of spermatozoa in their spermathecae. The sister queens reared by the grafting method were instrumentally inseminated with 8 μl fresh semen when they became 6 days old. One week after instrumental insemination, one batch of queens (2-week-old) was dissected for spermatozoa viability test. Another set of queens was introduced into production colonies in Langstroth hives after the onset of oviposition in the mating nuclei. The queens were maintained in production colonies for one year and two years until the dissection process for spermatozoa viability test. The viability of spermatozoa was measured by the dual staining method. We determined that the mean viability of spermatozoa in 2-week, 1-year and 2-year-old queens were 97.3%, 91.1% and 88.1%, respectively. The viability of spermatozoa in queens decreased with age, and the differences between the viability means were significant. However, we did not detect a steep decline in the viability of spermatozoa in queens (6% in one year and 9% in two years) in a wide range of timescale in contrast to previous reports. Furthermore, we found high viability of spermatozoa in the spermathecae of queens at the start of their lives and one- and two-years of age.

PDF

___

Al-Lawati H, Kamp G, Bienefeld K. 2009. Characteristics of the spermathecal contents of old and young honeybee queens. Journal of Insect Physiology, 55:117–122. doi: 10.1016/j.jinsphys.2008.10.01
Chaimanee V, Pettis JS. 2019. Gene expression, sperm viability, and queen (Apis mellifera) loss following pesticide exposure under laboratory and field conditions. Apidologie, 50:304– 316. doi: 10.1007/s13592-019-00645-4
Collins AM, Williams V, Evans JD. 2004. Sperm storage and antioxidative enzyme expression in the honey bee, Apis mellifera. Insect Molecular Biology, 13:141–146. doi: 10.1111/j.0962-1075.2004.00469.x
Gençer HV, Kahya Y. 2011. The viability of sperm in lateral oviducts and spermathecae of instrumentally inseminated and naturally mated honey bee (Apis mellifera L.) queens. Journal of Apicultural Research, 50:190–194. doi: 10.3896/IBRA.1.50.3.02
Gençer HV, Kahya Y, Woyke J. 2014. Why the viability of spermatozoa diminishes in the honeybee (Apis mellifera) within short time during natural mating and preparation for instrumental insemination. Apidologie, 45:757–770. doi: 10.1007/s13592-014-0295-0
Holman L. 2009. Sperm viability staining in ecology and evolution: potential pitfalls. Behavioral Ecology and Sociobiology, 63:1679–1688. doi: 10.1007/s00265-009-0816-4
Kahya Y, Gençer HV, Woyke J. 2008. Weight at emergence of honey bee (Apis mellifera caucasica) queens and its effect on live weights at the pre and post mating periods. Journal of Apicultural Research, 47:118–125. doi: 10.1080/00218839. 2008.11101437
Klenk M, Koeniger G, Koeniger N, Fasold H. 2004. Proteins in spermathecal gland secretion and spermathecal fluid and the properties of a 29 kDa protein in queens of Apis mellifera. Apidologie, 35:371–381. doi: 10.1051/apido:2004029
Laidlaw HH, Page RE. 1997. Queen rearing and bee breeding. Wiswcas Press, Cheshire, Connecticut, USA.:224
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. doi: 10.3389/fphys.2020.574856
Lodesani M, Balduzzi D, Galli A. 2004. A study on spermatozoa viability over time in honey bee (Apis mellifera ligustica) queen spermathecae. Journal of Apicultural Research, 43:27– 28. doi: 10.1080/00218839.2004.11101105
McAfee A, Chapman A, Pettis JS, Foster LJ, Tarpy DR. 2021a. Trade-offs between sperm viability and immune protein expression in honey bee queens (Apis mellifera). Communications Biology, 4:1–11. doi: 10.1038/s42003-020- 01586-w
McAfee A, Tarpy DR, Foster LJ. 2021b. Queen honey bees exhibit variable resilience to temperature stress. PLOS ONE, 16:e0255381. doi: 10.1371/journal.pone.0255381
Moritz RFA. 1984. The effect of different diluents on insemination success in the honeybee using mixed semen. Journal of Apicultural Research, 23:164–167. doi: 10.1080/00218839.1984.11100626
Pascini TV, Martins GF. 2017. The insect spermatheca: an overview. Zoology, 121:56–71. doi: 10.1016/j.zool .2016.12.001
Pettis JS, 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: 10.1371/journal.pone.0147220
Tarpy DR, Keller JJ, Caren JR, Delaney DA. 2012. Assessing the mating “health” of commercial honey bee queens. Journal of Economic Entomology, 105:20–25. doi: 10.1603/ec11276
Tarpy DR, Olivarez R. 2014. Measuring sperm viability over time in honey bee queens to determine patterns in stored-sperm and queen longevity. Journal of Apicultural Research, 53:493–495. doi: 10.3896/IBRA.1.53.4.02
Verma LR. 1973. An ionic basis for a possible mechanism of sperm survival in the spermatheca of the queen honey bee (Apis mellifera L.). Comparative Biochemistry and Physiology Part A: Physiology, 44:1325–1331. doi: 10.1016/0300-9629(73)90272-7
Verma LR. 1978. Biology of honeybee (Apis mellifera L.) spermatozoa .1. Effect of different diluents on motility and survival. Apidologie, 9:167–173. doi: 10.1051/apido: 19780301
Weirich GF, Collins AM, Williams VP. 2002. Antioxidant enzymes in the honey bee, Apis mellifera. Apidologie, 33:3– 14. doi: 10.1051/apido:2001001
Winston ML. 1987. The Biology of The Honey Bee. Cambridge, Mass: Harvard University Press.
Woyke J. 1962. Natural and artificial insemination of queen honeybees. Bee World, 43:21–25. doi: 10.1080/0005772X. 1962.11096922