Antibiotics used in animal agriculture are of increasing environmental concern due to the potential for antibiotic resistance in the environment following land application of manure. Yet, little is known about the impact of different manure application technologies on the environmental behavior of these antibiotics. Therefore, rainfall simulations were conducted to determine the fate and transport of pirlimycin, an antibiotic commonly used in dairy production. Three treatments (surface application, subsurface injection, and control with no manure) were setup in a randomized complete block design, using dairy manure spiked with pirlimycin. Rainfall simulations were conducted 0 and 7 days after application of manure. Surface runoff water samples were filtered, and water, sediment, and soil samples were analyzed using UPLC-MS/MS to determine antibiotic concentrations. Filtered water and sediment were analyzed to determine the amount of antibiotics associated with water and particulate matter in runoff. Soil samples were collected from all plots at two depths (0-5 cm and 5-20 cm), and soil samples were taken from injection slits and between slits in the plots receiving the injected treatment to determine spatial variability. Surface application resulted in 21 and 29 times the mass loss of pirlimycin in sediment and water from injection on day 0. If antibiotic transport is decreased using injection, subsurface injection could be used as a best management practice to increase nutrient capture and prevent loss of emerging contaminants to the environment.