Broiler chickens are fast growing monogastric animals considered a sensitive test species to evaluate the safety and nutritional equivalence between transgenic and non-transgenic grains as part of the human safety evaluation process. DAS-44406-6 soybean expresses three herbicide-tolerant proteins: the aryloxyalkanoate dioxygenase-12 (AAD-12) enzyme which provides tolerance to 2,4-dichlorophenoxyacetic acid (2,4-D) herbicides, the double-mutant 5-enolpyruvylshikimate-3-phosphate synthase (2mEPSPS) enzyme encoded by a modified version of the epsps gene from maize (Zea mays), which provides tolerance to glyphosate herbicides, and the phosphinothricin acetyltransferase (PAT) enzyme from Streptomyces viridochromogenes, which provides tolerance to glufosinate herbicides. A 42-day broiler study was conducted with diets containing toasted DAS-44406-6 soybean meal to evaluate nutritional wholesomeness and safety compared with non-transgenic, near-isoline soybean and conventional comparators. The study used a randomized complete block design with five dietary treatments assigned randomly within six blocks of 12 pens per treatment, with 10 birds per pen. Broiler performance and carcass parameters were measured over a 6-week period of exposure to diets containing different sources of toasted soybean meal and hulls. Among the 13 endpoints measured, only thigh weight was numerically different between birds fed DAS-44406-6 soybean meal and those fed isoline soybean meal. However, no significant differences to thigh weight were observed between birds fed DAS-44406-6 soybean and any of the non-transgenic reference varieties. Results indicate that DAS-44406-6 soybeans are nutritionally equivalent to conventional varieties.