We applied a screening-level phytotoxicity assay to evaluate the effects of 10 antibiotics (at concentrations ranging from 1 to 10,000 lg/L) on germination and early plant growth using three plant species: lettuce (Lactuca sativa), alfalfa (Medicago sativa), and carrot (Daucus carota). The range of phytotoxicity of the antibiotics was large, with EC 25 s ranging from 3.9 lg/L to [10,000 lg/L. Chlortetracycline, levofloxacin, and sulfamethoxazole were the most phytotoxic antibiotics. D. carota was the most sensitive plant species, often by an order of magnitude or more, followed by L. sativa and then M. sativa. Plant germination was insensitive to the antibiotics, with no significant decreases up to the highest treatment concentration of 10,000 lg/L. Compared with shoot and total length measurements, root elongation was consistently the most sensitive end point. Overall, there were few instances where measured soil concentrations, if available in the publicly accessible literature, would be expected to exceed the effect concentrations of the antibiotics evaluated in this study. The use of screening assays as part of a tiered approach for evaluating environmental impacts of antibiotics can provide insight into relative species sensitivity and serve as a basis by which to screen the potential for toxic effects of novel compounds to plants.