Runoff generated from livestock manure amended row crop fields is, one of the major pathways of hormone transport to the aquatic environment. The study determined the effects of manure handling, tillage methods, and rainfall timing on the occurrence and transport of steroid hormones in runoff from the row crop field. Stockpiled and composted manure from hormone treated and untreated animals were applied to test plots and subjected to two rainfall simulation events 30 days apart During the two rainfall simulation events, detection of any steroid hormone or metabolites was identified in 8-86% of runoff samples from any tillage and manure treatment The most commonly detected hormones were 17 beta-estradiol, estrone, estriol, testosterone, and alpha-zearalenol at concentrations ranging up to 100-200 ng L-1. Considering the maximum detected concentrations in runoff, no more than 10% of the applied hormone can be transported through the dissolved phase of runoff. Results from the study indicate that hormones can persist in soils receiving livestock manure over an extended period of time and the dissolved phase of hormone in runoff is not the preferred pathway of transport from the manure applied, fields irrespective of tillage treatments and timing of rainfall. (C) 2016 Elsevier B.V. All rights reserved.