In regions of concentrated poultry production, poultry litter (PL) that contains significant quantities of trace elements is commonly surface-applied to pastures at high levels over multiple years. This study examined the effect of long-term applications of PL on soil concentrations of arsenic (As), copper (Cu), Zinc (Zn), and the uptake of these elements by bermuda grass grown on Cecil (well-drained) and Sedgefield (somewhat poorly-drained) soils. The results showed that concentrations of As, Cu, and Zn in soils that had received surface-applied PL over a 14-year period were significantly greater than untreated soil at 0–2.5 and 2.5–7.5 cm depths. However, the levels were well below the USEPA loading limits established for municipal biosolids. Arsenic fractionation showed that concentrations of all As fractions were significantly greater in PL-amended soils compared to untreated soils at 0–2.5 and 2.5–7.5 cm depths. The residual fraction was the predominant form of As in all soils. The water-soluble and NaHCO3
-associated As were only 2% of the total As. Significant differences were found in concentrations of these trace elements and phosphorus (P) in forage from PL-amended soils compared to that in untreated plots. The concentrations of Cu, Zn, As, and P were significantly greater in forage from Sedgefield amended soil compared to Cecil soil, but were in all cases below levels of environmental concern.