Using the washing technique for semen collection: ‑ discussion and evaluation of Apis mellifera L. spermatozoa concentration in four different layers formed after dilution and centrifugation of semen in tris‑buffer diluent

A quick review of the progress made throughout this century in Apis semen knowledge, and its relationship with the development made in instrumental insemination techniques, is presented. The relative (dis)advantages of the washing technique, in comparison to the traditional methods used for semen collection, are considered. Also, the major problems still to overcome in order to improve the washing technique feasibility are addressed. The main results obtained in this work show that there are statistically significant differences in sperm number and concentration among the four different layers studied, although the two top layers formed within the capillary tubes did not statistically differ between themselves (as indeed also happened regarding the two bottom ones). On the other hand, it was found that the two top layers were significantly poorer in sperm number and concentration than the two bottom ones. Clearly, there is a relationship between sperm concentration and the position it occupies in the sperm column. The washing technique seems to be more time‑ efficient than the traditional semen collection methodology. The potential value of former technique is discussed.

Inseminação Instrumental; Espermatozóides, Abelhas

Washing technique; Instrumental insemination; Spermatozoa; Honey bees