Our research presents a proof-of-concept that explores a new and innovative method to identify large animals in aerial imagery with single day image differencing. We acquired two aerial images of eight fenced pastures and conducted a principal component analysis of each image. We then subtracted the first principal component of the two pasture images followed by heuristic thresholding to generate polygons. The number of polygons represented the number of potential cattle (Bos taurus) and horses (Equus caballus) in the pasture. The process was considered semi-automated because we were not able to automate the identification of spatial or spectral thresholding values. Imagery was acquired concurrently with ground counts of animal numbers. Across the eight pastures, 82% of the animals were correctly identified, mean percent commission was 53%, and mean percent omission was 18%. The high commission error was due to small mis-alignments generated from image-to-image registration, misidentified shadows, and grouping behavior of animals. The high probability of correctly identifying animals suggests short time interval image differencing could provide a new technique to enumerate wild ungulates occupying grassland ecosystems, especially in isolated or difficult to access areas. To our knowledge, this was the first attempt to use standard change detection techniques to identify and enumerate large ungulates.