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Vertebrate reproductive science and technology
RESEARCH ARTICLE

Gel-coated tubes extend above-freezing storage of honey bee (Apis mellifera) semen to 439 days with production of fertilised offspring

Brandon K. Hopkins A C , Susan W. Cobey A , Charles Herr B and Walter S. Sheppard A
+ Author Affiliations
- Author Affiliations

A Department of Entomology, Washington State University, 100 Dairy Road, Pullman, WA 99164, USA.

B Center for Animals Near Biological Extinction, Department of Biology, Eastern Washington University, 7210 W Melville Road, Cheney, WA 99004, USA.

C Corresponding author. Email: bhopkins@wsu.edu

Reproduction, Fertility and Development 29(10) 1944-1949 https://doi.org/10.1071/RD16087
Submitted: 12 March 2016  Accepted: 6 November 2016   Published: 16 December 2016

Abstract

Honey bees are an important agricultural species; however, relatively little work has been done to improve artificial reproductive technologies for this animal. The collection and distribution of germplasm for breeding and conservation is critical for improving managed honey bee populations and conserving threatened subspecies. The most efficient method of controlling breeding in honey bees is by artificial insemination. The collection of semen for insemination requires the use of antibiotics, which is especially critical if semen is to be stored for any length of time. The introduction of antibiotics is normally done through a balanced salt solution. In this study we compare, at two temperatures, the storage of undiluted semen in antibiotic–gel-coated capillary tubes with storage of semen diluted in a balanced salt solution containing antibiotics. Live–dead cell staining and artificial insemination of honey bee queens were performed at 45, 99 and 439 days after collection of the semen. In every case the antibiotic–gel-coated tube storage method at 14°C produced a higher percentage of fertilised offspring. This study demonstrates the longest period of time spermatozoa have been stored above freezing while maintaining fertilisation capacity.

Additional keywords: instrumental insemination, in vitro, long-term storage.


References

Büchler, R., Berg, S., and Le Conte, Y. (2010). Breeding for resistance to Varroa destructor in Europe. Apidologie (Celle) 41, 393–408.
Breeding for resistance to Varroa destructor in Europe.Crossref | GoogleScholarGoogle Scholar |

Cobey, S. W. (2007). Comparison studies of instrumentally inseminated and naturally mated honey bee queens and factors affecting their performance. Apidologie (Celle) 38, 390–410.
Comparison studies of instrumentally inseminated and naturally mated honey bee queens and factors affecting their performance.Crossref | GoogleScholarGoogle Scholar |

Collins, A. M. (2000). Survival of honey bee (Hymenoptera: Apidae) spermatozoa stored at above-freezing temperatures. J. Econ. Entomol. 93, 568–571.
Survival of honey bee (Hymenoptera: Apidae) spermatozoa stored at above-freezing temperatures.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD3cvhtlaltQ%3D%3D&md5=e457cd7740e86b255ef3a539ad6132aeCAS |

Collins, A. M., and Donoghue, A. M. (1999). Viability assessment of honey bee, Apis mellifera sperm using dual fluorescent staining. Theriogenology 51, 1513–1523.
Viability assessment of honey bee, Apis mellifera sperm using dual fluorescent staining.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD3c7ptFChsA%3D%3D&md5=f3f669f738a5431fcce22aefc219eb05CAS |

Douard, V., Hermier, D., and Blesbois, E. (2000). Changes in turkey semen lipids during liquid in vitro storage. Biol. Reprod. 63, 1450–1456.
Changes in turkey semen lipids during liquid in vitro storage.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXnslCkt7s%3D&md5=803dc9c6562653ff1d683a18da7d6516CAS |

Gallai, N., Salles, J., Settele, J., and Vaissiere, B. E. (2009). Economic valuation of the vulnerability of world agriculture confronted with pollinator decline. Ecol. Econ. 68, 810–821.
Economic valuation of the vulnerability of world agriculture confronted with pollinator decline.Crossref | GoogleScholarGoogle Scholar |

Harbo, J. R. (1974). A technique for handling stored semen of honey bees. Ann. Entomol. Soc. Am. 67, 191–194.
A technique for handling stored semen of honey bees.Crossref | GoogleScholarGoogle Scholar |

Harbo, J. R. (1985). Instrumental insemination of queen bees-1985. Part 1. Am. Bee J. 125, 197–202.

Hopkins, B. K., and Herr, C. (2010). Factors affecting the successful cryopreservation of honey bee (Apis mellifera) spermatozoa. Apidologie (Celle) 41, 548–556.
Factors affecting the successful cryopreservation of honey bee (Apis mellifera) spermatozoa.Crossref | GoogleScholarGoogle Scholar |

Hopkins, B. K., Herr, C., and Sheppard, W. S. (2012). Sequential generations of honey bee (Apis mellifera) queens produced using cryopreserved semen. Reprod. Fertil. Dev. 24, 1079–1083.
Sequential generations of honey bee (Apis mellifera) queens produced using cryopreserved semen.Crossref | GoogleScholarGoogle Scholar |

Ibrahim, A., Reuter, G. S., and Spivak, M. (2007). Field trial of honey bee colonies bred for mechanisms of resistance against Varroa destructor. Apidologie (Celle) 38, 67–76.
Field trial of honey bee colonies bred for mechanisms of resistance against Varroa destructor.Crossref | GoogleScholarGoogle Scholar |

Johnson, L. A., Weitze, K. F., and Maxwell, W. M. C. (2000). Storage of boar semen. Anim. Reprod. Sci. 62, 143–172.
Storage of boar semen.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXlt1Kgsbc%3D&md5=c7a7e93123fa559533ce3650cc83c6e3CAS |

Leibo, S. P., Semple, M. E., and Kroetsch, T. G. (1994). In vitro fertilization of oocytes by 37-year-old cryopreserved bovine spermatozoa. Theriogenology 42, 1257–1262.
In vitro fertilization of oocytes by 37-year-old cryopreserved bovine spermatozoa.Crossref | GoogleScholarGoogle Scholar |

Locke, S. J., and Peng, Y. S. (1993). The effects of drone age, semen storage and contamination on semen quality in the honey bee (Apis mellifera). Physiol. Entomol. 18, 144–148.
The effects of drone age, semen storage and contamination on semen quality in the honey bee (Apis mellifera).Crossref | GoogleScholarGoogle Scholar |

Mazur, P. (1984). Freezing of living cells: mechanisms and implications. Am. J. Physiol. 247, 125–142.

Mendenhall, W. (1979) ‘Introduction to Probability and Statistics’. 5th edn. (Duxbury Press: Belmont, CA, USA.)

Poole, H. K., and Taber, S. (1970). In vitro preservation of honey bee semen enhanced by storage at 13–15°C. Ann. Entomol. Soc. Am. 63, 1673–1674.
In vitro preservation of honey bee semen enhanced by storage at 13–15°C.Crossref | GoogleScholarGoogle Scholar |

Rinderer, T., Harris, J., Hunt, G., and de Guzman, L. (2010). Breeding for resistance to Varroa destructor in North America. Apidologie (Celle) 41, 409–424.
Breeding for resistance to Varroa destructor in North America.Crossref | GoogleScholarGoogle Scholar |

Salamon, S., and Maxwell, W. M. C. (1995). Frozen storage of ram semen. 2. Causes of low fertility after cervical insemination and methods of improvement. Anim. Reprod. Sci. 38, 1–36.
Frozen storage of ram semen. 2. Causes of low fertility after cervical insemination and methods of improvement.Crossref | GoogleScholarGoogle Scholar |

Taber, S., and Blum, M. (1960). Preservation of honey bee semen. Science 131, 1734–1735.
Preservation of honey bee semen.Crossref | GoogleScholarGoogle Scholar |

Taber, S., Poole, H. K., and Edwards, J. F. (1979). Enhanced fertility of honey bee semen stored in vitro and possibly a reversal of senescence. Apidologie (Celle) 10, 129–136.
Enhanced fertility of honey bee semen stored in vitro and possibly a reversal of senescence.Crossref | GoogleScholarGoogle Scholar |

Taylor, M. A., Guzman-Novoa, E., Morfin, N., and 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.
Improving viability of cryopreserved honey bee (Apis mellifera L.) sperm with selected diluents, cryoprotectants, and semen dilution ratios.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXnsFSjtrw%3D&md5=d148039eed29bf23c8b83e81470b9c98CAS |

Trzcińska, M., Bryla, M., and Smorag, Z. (2011). Apoptotic-like changes in the spermatozoa of fresh and stored boar semen and the quality of embryos produced in vivo. Anim. Reprod. Sci. 124, 90–97.
Apoptotic-like changes in the spermatozoa of fresh and stored boar semen and the quality of embryos produced in vivo.Crossref | GoogleScholarGoogle Scholar |

Verberckmoes, S., Van Soom, A., Dewulf, J., and de Kruif, A. (2005). Comparison of three diluents for the storage of fresh bovine semen. Theriogenology 63, 912–922.
Comparison of three diluents for the storage of fresh bovine semen.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXjsVWq&md5=6abdda7f389cf522cb60b27cf438ee36CAS |

Verma, L. (1978). Biology of honey bee (Apis meellifera L.) spermatozoa. 1. Effects of different diluents on motility and survival. Apidologie (Celle) 9, 167–174.
Biology of honey bee (Apis meellifera L.) spermatozoa. 1. Effects of different diluents on motility and survival.Crossref | GoogleScholarGoogle Scholar |