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Standard methods for instrumental insemination of Apis mellifera queens

Summary:

Honey bee queens are highly polyandrous and mate in flight. Instrumental insemination is an essential tool that provides complete control of honey bee mating for research and breeding purposes. The technique requires specialized equipment to anesthetize and immobilize the queen and to collect and deliver semen from the drones. Semen is harvested from mature drones by hand eversion of the endophallus and collected into a syringe. The queen is placed in a chamber and anesthetized during the procedure of insertion of semen into the oviducts. Queens are introduced into colonies and their performance can equal to that of naturally mated queens, given proper technique and care.

    1. 1.1. Applications
    2. 1.2. Current uses
    1. 2.1. Instrument options and choices
    2. 2.2. Saline diluent formulas
    3. 2.2.1. Simple saline formula
    4. 2.2.2. Formula for semen storage and mixing
    1. 3.1. Eversion of the endophallus
    2. 3.2. Semen collection
    3. 3.3. Insemination of the queen
    4. 3.4. Field dissection of the spermatheca
    1. 4.1. Maintenance of drones for instrumental insemination
    2. 4.2. Maintenance of queens before and after insemination
    3. 4.3. Factors influencing results of instrumental insemination
    1. 5.1. Homogenizing honey bee semen
    2. 5.2. Short term semen storage at above freezing temperatures
    3. 5.2.1. To seal semen filled capillary tubes
    4. 5.2.2. To remove the petroleum seal
    5. 5.3. Cryopreservation of semen
    6. 5.4. Techniques for counting sperm
    7. 5.4.1. Queen spermathecae
    8. 5.4.2. Drone seminal vesicles
    9. 5.4.3. Sperm count protocol (unstained sperm)
    10. 5.5. Sperm viability measures
    11. 5.5.1. Reagents
    12. 5.5.2. Equipment
    13. 5.5.3. Procedure