Hygienic behavior in honey bees is a heritable trait of individual workers that confers colony-level resistance against various brood diseases. Hygienic workers detect and remove dead or diseased brood from sealed cells. However, this behavior is quite rare, with only c.10% of unselected colonies showing high levels of hygiene. Beekeepers can potentially increase this by screening colonies for hygiene and breeding from the best. However, the level of hygiene expressed by a colony is variable, which poses a challenge to colony selection. In this study, we systematically varied two factors thought to be of importance in influencing hygiene levels, "nectar" availability, by feeding or not feeding sucrose syrup, and brood amount, by adding or removing brood, to determine what effect they had on hygienic behavior. We tested 19 colonies repeatedly over a 4-mo period using the freeze-killed brood assay, a standard technique to quantify hygienic behavior. Two days after freeze-killed brood treatment, our colonies showed a wide range of brood removal levels, with colony means ranging from 31.7 +/- 22.5 to 93 +/- 6.9 (mean % +/- SD). Neither the food nor the brood manipulation had an effect on hygiene levels. Colony size and time of year were also nonsignificant. The only significant effect was a three-way interaction between syrup availability, amount of brood, and time of the year, resulting in reduced hygienic behavior early in the season (spring), in colonies with added brood that were not fed sucrose syrup. Overall, these results suggest that hygienic behavior is not greatly affected by environmental conditions typical of a real-life beekeeping, and that screening of colonies can be done anytime without special regard to nectar conditions or brood levels.