The Effect of Semen Dilution on the Number of Spermatozoa Entering the Spermatheca of the Queen Honey Bee (Apis mellifera L.)

This study aimed to determine whether the addition of diluent to fresh semen before injection affects the number of spermatozoa entering the spermatheca of the instrumentally inseminated queens. The queens reared by the grafting method were introduced into mating nucs one day before adult emergence. When 7-day old, the queens were allocated into four groups for the following instrumental insemination treatments. The queens in group 1 and group 3 were inseminated with 4 μl and 8 μl fresh semen, respectively (FS - 4 μl and FS - 8 μl). In group 2, the queens were inseminated with 4 μl fresh semen diluted 1-fold with NaCl solution (DS - 8 μl). In group 4, the queens were inseminated with 8 μl fresh semen diluted 1-fold with NaCl solution (DS - 16 μl). Each queen was dissected after the onset of oviposition for counting the number of spermatozoa in the spermatheca. There was not a significant difference in spermatozoa number between queens inseminated with 4 μl FS (2.57 million) and 8 μl DS (2.68 million). The queens inseminated with 16 μl DS (3.51 million) had fewer spermatozoa than the queens inseminated with 8 μl FS (4.46 million). The queens inseminated with 16 μl DS had more spermatozoa (3.51 million) than the queens inseminated with 4 μl FS (2.57 million, and 8 μl DS (2.68 million). The results revealed that the addition of 1-fold diluent to 4 μl fresh semen did not alter the number of spermatozoa. In contrast, adding 1-fold diluent to 8 μl fresh semen adversely affected the number of spermatozoa entering the spermatheca.

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Arslan S, Cengiz M M, Gül A, Sayed, S. 2021. Evaluation of the standards compliance of the queen bees reared in the Mediterranean region in Turkey. Saudi Journal of Biological Sciences 28(5): 2686-2691. DOI: 10.1016/j.sjbs.2021.03.009
Cobey SW. 2007. Comparison studies of instrumentally inseminated and naturally mated honey bee queens and factors affecting their performance. Apidologie 38:390–410. DOI: 10.1051/apido:2007029
Delaney D, Keller J, Caren J, Tarpy D. 2010. The physical, insemination, and reproductive quality of honey bee queens (Apis mellifera L.). Apidologie 42:1–13. DOI: 10.1051/apido/2010027
Gąbka J, Cobey SW. 2018. Factors, based on common practices, affecting the results of instrumental insemination of honey bee queens. Apidologie 49:773–780. DOI: 10.1007/s13592- 018-0606-y
Gençer HV, Firatli C. 2005. Reproductive and morphological comparisons of drones reared in queenright and laying worker colonies. Journal of Apicultural Research 44:163–167
Gençer HV, Kahya Y. 2011a. 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
Gençer HV, Kahya Y. 2011b. Are sperm traits of drones (Apis mellifera L.) from laying worker colonies noteworthy? Journal of Apicultural Research 50:130–137. DOI: 10.3896/IBRA.1.50.2.04
Harbo JR. 1977. Survival of honey bee spermatozoa in liquid nitrogen. Annals of the Entomological Society of America 70:257–258. DOI: 10.1093/aesa/70.2.257
Harbo JR. 1983. Survival of honey bee (Hymenoptera: Apidae) spermatozoa after two years in Liquid Nitrogen (–196°C). Annals of the Entomological Society of America 76:890–891. DOI: 10.1093/aesa/76.5.890
Hatjina F. 2012. Greek honey bee queen quality certification. Bee World 89:18–20
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
Koeniger N, Koeniger G. 1991. An evolutionary approach to mating behaviour and drone copulatory organs in Apis. Apidologie 22:581–590. DOI: 10.1051/apido:19910602
Laidlaw HH, Page RE. 1997. Queen Rearing and Bee Breeding. Wiswcas Press, Cheshire, Connecticut, USA.:224
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: 10.4081/ijas.2004.385
Mackensen O. 1947. Effect of carbon dioxide on initial oviposition of artificially inseminated and virgin queen bees. Journal of Economic Entomology 40:344–349
Mackensen O. 1964. Relation of semen volume to success in artificial insemination of queen honey bees. Journal of Economic Entomology 57:581–583. DOI: 10.1093/jee/57.4.581
Mackensen O. 1969. Effect of diluents and temperature on instrumental insemination of queen honey bees. Journal of Economic Entomology 62:1370–1372
Mackensen O, Roberts WC. 1948. A Manual for the Artificial Insemination of Queen Bees. USDA, Agricultural Research Administration, Bureau
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
Skowronek W, Kruk C, Loc K. 1995. The insemination of queen honeybees with diluted semen. Apidologie 26:487–493. DOI: 10.1051/apido:19950605
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
Woyke J. 1962. Natural and artificial insemination of queen honeybees. Bee World 43:21–25. DOI: 10.1080/0005772X.1962.11096922
Woyke J. 1983. Dynamics of entry of spermatozoa into the spermatheca of instrumentally inseminated queen honeybees. Journal of Apicultural Research 22:150–154