Table 1.

Effects of different copulation factors on queen post-mating changes. Shown are outcomes of different treatments on queen post-mating changes, worker behavior, and colony longevity. Carbon dioxide (CO₂) indicates queens that were exposed to CO₂ alone but were not instrumentally inseminated. Virgins are control queens that were handled similarly to the other treatments, but were not inseminated or treated with any substance. CPM indicates queens that were both exposed to CO₂ and physically manipulated in the oviduct to simulate the physical aspects of instrumental insemination. SDI are queens that were inseminated with semen from one drone (~1 μL) and MDI are queens that were inseminated with semen from multiple drones (~10 μL). The outcome of inseminating queens with 8 μL versus 1 μL semen is also shown. Saline serves as an insemination and volume control for semen in order to test the effects of semen components on post-mating changes. Lastly, seminal fluid is a component of semen that contains proteins (SFPs) that are likely important for inducing post-mating changes and Hayes solution is often used as a semen and seminal fluid diluent. “Yes, ns” in the table indicates that results trended toward the respective phenotype, but they were not statistically significant.

Mating/Insemination Factors
Queen Post-Mating Outcomes	CO2 vs. Virgins	CPM vs. Virgins	SDI and MDI vs. Virgins	SDI vs. MDI	Insemination Volume: 8 μL vs. 1 μL	Semen vs. Saline	Seminal Fluid vs. Hayes	Naturally Mated vs. Virgin
Reduced sexual receptivity?	Yes [99]	Yes [93]	unknown	unknown	Yes, ns [58]	Yes, ns [55]	Yes [93]	Yes [13,63,68]
Greater ovary activation?	Yes, ns [93]; Yes [96,99]	Yes [93]	unknown	unknown	Yes [58]	Yes [55]	unknown	Yes [72,73,96]
Enhanced worker retinue response?	Yes [93]	No [93]	Yes [47,48]	Yes [47,48]	Yes [50,51]	Yes [51] Yes, ns [49]	Yes [93]	Yes [47,49,67]
Modulated Mandibular gland pheromone production?	Yes [93]	Yes [93]	Yes [47]	Yes [47]	Yes [51]	Yes [51]	unknown	Yes [49,54,55,84]
Modulated Dufour’s gland pheromone production?	No [93]	No [93]	Yes [48]	Yes [48]	No Difference [51]	No Difference [51]	unknown	Yes [65]
# genes differentially expressed in brain out of all transcripts that were detected	234/9091 [93]	504/9091 [93]	unknown	unknown	unknown	44/9850 [68]	unknown	576/10,468 [57] 180/9850 [68]
# genes differentially expressed in ovaries out of all transcripts that were detected	unknown	unknown	unknown	unknown	unknown	unknown	unknown	217/7377 [57] regulation of biogenic amine receptor genes [96]