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Differences over time in the quality of semen present in the honey bee (Apis mellifera ligustica) queen spermatheca were<br />studied. An increase in the non-vital spermatozoa was shown to be evident (P>0.05) between the 12th and 24th month.<br />The study of semen viability demonstrated that the passage of the semen to the spermatheca is due to sperm motility.<br />In the queen inseminated with non-viable spermatozoa, no semen was detected in the spermatheca. Queens inseminated<br />twice with a Hyes solution/semen mixture (1:1) stored as many spermatozoa in their spermatheca as those inseminated<br />once with the classic technique. Queen replacement, oviposition and other functional characteristics were similar<br />to those observed in the classic insemination procedure.