Honey bee samples, 5 bees per colony each time-point, were homogenized in 1 mL 50% TRIzol Reagent (Sigma) and 50% phosphate buffered saline (PBS, UCSF Cell Culture) solution in a 2 mL micro-centrifuge tube containing one sterile zinc-coated steel ball bearing (5 mm) using a TissueLyzer II (Retsch), for 4 minutes at 30 Hz. RNA was isolated according to TRIzol Reagent (Invitrogen) manufacturer's instructions. In brief, TRIzol reagent honey bee homogenate was combined with 0.1 ml chloroform and mixed by vortexing for 5 seconds, samples were incubated at room temperature for 5 minutes, prior to centrifugation for 10 minutes at 13,2006g in a table top centrifuge. Next, 700 mL of the aqueous phase was transferred to a new microfuge tube containing 490 mL isopropanol. Following mixing, the samples were incubated at 220uC for 20 minutes and then either centrifuged (13, 2006 g for 15 min) or further purified utilizing Zymo-III RNA columns according to manufacture's instructions (Zymo). RNA was extracted from five bees collected from the colony entrance for each of the time-course samples.