Olfactory receptors (ORs) are important for insects to recognize and discriminate odorants in the environment and are mainly expressed in olfactory and gustatory organs. Little is known about the potential OR functions in non-olfactory tissues. In the present study, we evaluated the possibility of odorant receptors AcerOr1 and AcerOr2 (AcerOr2 is orthologous to the co-receptor) mediating sperm DNA integrity, and the relationship between sperm DNA integrity and semen parameters in Apis cerana cerana. Based on previous findings in mammals, we speculated that the Ca2+/calmodulin (CaM)/CaM-dependent protein kinase II (CaMKII) signaling pathway might be involved in the regulation of sperm motility in A. cerana cerana. The results showed that both AcerOr1 and AcerOr2 are expressed in the sperms and testis, that components associated with the putative Ca2+/CaM/CaMKII signaling pathway are present in A. cerana cerana sperms, and that at least CaM and CaMKII are localized in the sperms and testis. The AcerOr2 agonist VUAA1 significantly improved sperm motility parameters and apoptosis of sperm cells effect DNA integrity, whereas the CaM inhibitor W7 decreased sperm motility parameters and apoptosis of sperm cells, which affects DNA integrity. We also found a positive correlation between sperm DNA integrity and semen quality. These results indirectly as well as directly suggest that OR-mediated sperm responses and the Ca2+/CaM/CaMKII signaling pathway might affect semen quality and might be useful in regulating insect reproduction in future.