Dissertation
ARTIFICAL REPRODUCTIVE TECHNIQUES IN HONEY BEES: SPERM CELL PHYSIOLOGY, SEMEN COLLECTION AND STORAGE
Doctor of Philosophy (PhD), Washington State University
01/2014
Handle:
https://hdl.handle.net/2376/117726
Abstract
Modern agriculture in much of the world is geared toward large acreages of monoculture to maximize food production for an ever-increasing demand. The only highly polylectic species that is capable of pollinating a wide variety of crops and also being managed on such a large commercial scale is the western honey bee (Apis mellifera). Given the importance of honey bees, it is concerning that the agricultural industry has lagged in techniques and technologies that have propelled the rapid advancement of desirable traits in other agricultural species. We are attempting to improve the current state of artificial reproductive techniques (ART) for honey bees. Our work examined sperm cell metabolic energy pathways to better understand how to develop improved semen collection and storage practices. We found that honey bee sperm cells do not use oxidative phosphorylation and are reliant on glycolysis for energy metabolism. With this we tested the long-standing practice of using sugars in saline solution for the collection and instrumental insemination (II) of queens, finding that presence or absence of sugars in the diluent has no effect on the number of spermatozoa present in the queens' spermatheca following II. To further improve collection and insemination there was a need to improve the diluent used for collection and to improve storage conditions. We examined buffers and antibiotics for free radical production under laboratory lighting conditions, selected a suite that had the lowest free radical production, and tested those at different concentrations for cytotoxicity. Above-freezing storage of semen was tested including a diluent-free storage system in which semen was stored and capable of fertilizing ova after 439 days. We demonstrated the potential for sequential backcrosses using cryopreserved semen, and then collected semen from Old World populations in Italy, Slovenia, and Republic of Georgia. Collections of cryopreserved semen in the Republic of Georgia are being used to reintroduce the subspecies A.m. caucasica into the U.S., during this process we show the stability of cryopreserved semen during the two-year period it has been frozen.
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Details
- Title
- ARTIFICAL REPRODUCTIVE TECHNIQUES IN HONEY BEES: SPERM CELL PHYSIOLOGY, SEMEN COLLECTION AND STORAGE
- Creators
- Brandon Kingsley Hopkins
- Contributors
- Walter S Sheppard (Advisor)David W Crowder (Committee Member)Gary H Thorgaard (Committee Member)Chales M Herr (Committee Member)Carol M Anelli (Committee Member)
- Awarding Institution
- Washington State University
- Academic Unit
- Entomology, Department of
- Theses and Dissertations
- Doctor of Philosophy (PhD), Washington State University
- Number of pages
- 108
- Identifiers
- 99900581737201842
- Language
- English
- Resource Type
- Dissertation