Effect of chamber characteristics, incubation, and diluent on motility of honey bee (Apis mellifera) drone sperm
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In this study, we compared the effect of five different sample viewing devices (slide coverslips, Makler, Leja10, Leja20, and ISAS10 chambers), incubation time, analysis time, microscopic field analysis, and diluent used on honey bee semen motility parameters. Using media without proteins, a lower proportion of total motile and of freely motile sperm (those non-adhering to the glass surface) were observed for slide coverslip and slide coverslip–Makler chambers, respectively, than in other chambers, while the percentage of circular sperm followed an opposite trend. Significant increases in all motility parameters were observed when loaded Leja10 chambers were maintained at 35 °C. During microscopic field analysis in the Leja Chamber, the percentage of freely motile sperm decreased and those of circular sperm increased in the last fields evaluated. The addition of 2% of BSA to the diluent clearly reduced the sperm adhesion to glass surface when using slide coverslip and Makler chambers. This study confirms that the choice of chamber and diluent used to assess honey bee drone sperm motility has a significant effect on the results wherein traditional slide coverslips are contraindicated.
KeywordsApis mellifera iberiensis sperm motility
JY conceived this research, designed and performed experiments and analysis, and wrote the paper; IP and PS participated in the design, the experiments and participated in the revisions of it.
This work was supported by the Spanish MINECO (grant AGL2017-85030-R), and the DGA-FSE (grant A07_17R).
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