Honey thinners for cryopreservation of honey bee drones semen

Abstract

A preliminary test of two variants of the honey diluent, PP Pechnikov and PN Skatkin, for cryopreservation of the sperm of honey bee drones was carried out. The first option: honey 10% - 50 ml, lactose - 10 mg, sucrose 10 mg, egg yolk - 2.5 ml, glycerin 3% - 6 ml 250 μl. In the second variant, glycerin is replaced by DMSO 10% in a volume of 5 ml. A 10% working solution of honey was prepared in deionized water according to the method of P.P. Pechnikov and P.N. Skatkin on the basis of honey from white acacia. In a 1.8 ml Nunc cryotube, 80 μl of freshly prepared diluent was added, then 10 μl of semen, and everything was mixed. The samples prepared in this way were placed in visotubes, which, in turn, were immersed in a vessel with water at room temperature (24-26 ° C). After that, this vessel was placed in a refrigerator for 2 h at 3 ° C for equilibration. The cryostorage period was one day.The samples were thawed in a water bath at 37 ° С for 1 min. After thawing, 1 ml of a diluent heated to 37 ° C was added to the sample, carefully pipetted and left for 5-10 min. Then, a single centrifugation was carried out for 3 min at 3000 rpm. After centrifugation, the supernatant was removed and sperm was collected from the bottom of the cryovial using the capillary of the syringe block for uterus.The results of the analysis showed that the most optimal option for artificial insemination of queen bees is the option with DMSO. A single insemination with a volume of injected semen of 8-10 μL was used. For insemination, we used infertile queens aged 7–8 days. The printed brood was counted by direct counting of the number of bee and drone cells.It was found that natural bee honey from white acacia showed the properties of a cryoprotectant, which in combination with egg yolk and DMSO made it possible to obtain brood of working bees over 50%.