6% yeast extract (TSAYE) to obtain a uniform lawn. After 24 h of incubation at 35 ± 2 °C, the bacterial lawn was harvested in 10 ml of sterile 0.1% peptone water (Difco), which was then added to 30 ml of GSK2118436 cell line 0.1% peptone water. Thereafter, 15 ml of culture was mixed with 150 g of each nut type in a sterile Whirlpak® filter bag for 1 min to give a target inoculum of ~ 108 CFU/g, after which the nuts were poured onto a raised aluminum mesh rack and dried in a biosafety hood at an air flow of ~ 0.56 m/s

for 20 min to remove excess peptone water. Thereafter, the inoculated samples were transferred to a glove box (EW-34788-00, Cole-Parmer, Vernon Hills, IL) for subsequent water activity (aw) conditioning. Four saturated salt solutions — CH3COOK, K2CO3, NaNO2, and KCl, were used to condition the nuts to aw values of 0.23, 0.45, 0.64, and 0.84 at 20 °C, respectively. The lid of a

steel tray was modified by installing a small fan and inlet/outlet holes to enhance air circulation inside the glove box. The tray was filled with 150–250 g of the appropriate salt and then saturated with de-ionized water. The conditioning salt tray, inoculated nut samples, a water activity meter (Hygrolab 3, Rotronic Instrument Corp., Hauppauge, NY), a digital relative humidity/temperature SAHA HDAC in vitro meter (pre-installed in the glove box), and Whirl-Pak® sample bags (4 oz) (Nasco, Fort Atkinson, WI) were then placed in the glove box, after which the main door was closed for further conditioning. To monitor the conditioning process, tightly sealed Petri dishes (10 mm × 40 mm diam.) containing ~ 10 g of each nut type were removed from the most glove box through a pass box door that maintained a closed system for the sample and the glove box. Conditioning

to equilibrium moisture content (EMC) (< 0.03% weight change over ~ 24 h) usually took about 6–7 days. After reaching equilibrium, ~ 5 g of the conditioned nuts was transferred to a sterile Whirl-Pak® sample bag in the conditioning glove box, in order to maintain the established humidity around the sample. Final EMC was measured using an oven drying method, and aw was measured using the water activity meter on the day of irradiation. The inoculated aw-conditioned samples (5 g, ~ 5 nuts) were irradiated in a prototype X-ray irradiator (Rainbow™ II, Rayfresh Foods Inc., Ann Arbor, MI), which currently is housed in the biosafety level-2 pilot plant at Michigan State University. The irradiator consists of an industrial grade X-ray tube (modified OEG-75, Varian Medical System, Salt Lake City, UT), high voltage source, and cooling unit. The X-ray tube operates at a maximum constant potential of 70 kV and a filament current of 57 mA, which gives 4 kW of maximum allowable input power. Five different surface doses (0.3–5.