The spider Latrodectus hesperus Chamberlin & Ivie (Araneae: Theridiidae) was subjected to low and ultralow oxygen (ULO) treatments at different temperatures. Complete control of the spiders was achieved in 24-h ULO treatments with 0.5% O₂ or lower at 1°C and in a 24-h low oxygen (2%) treatment at 15°C. Oxygen level and temperature greatly affected spider mortality. At 1°C, as oxygen level was decreased from 2 to 0.5%, spider mortality increased from 0 to 100%. At 2% O₂, as temperature was increased from 1 to 15°C, spider mortality increased from 0 to 100%. Grape clusters from two table grape (Vitis spp.) cultivars, ‘Thompson Seedless’ and ‘Flame Seedless’, were subjected to the 24-h ULO treatment with 0.5% O₂ at 1°C. The ULO treatment had no negative effects on grape quality. Because of the relatively short treatment time, effectiveness at low storage temperature and the easily attained oxygen level, we conclude that the ULO treatment have good potential to be implemented commercially for control of black widow spiders on harvested table grapes.