AbstractA new test design for the non‐axenic submergent aquatic macrophytes Elodea canadensis Michx. and Myriophyllum spicatum L. has been developed for potential use in herbicide toxicity testing. For the non‐axenic cultures, the best growth conditions were observed in the Elendt‐M4 medium in which no growth of algae or bacteria was observed. Cuttings were placed in beakers containing only the artificial M4 medium or were planted in small beakers containing OECD (Organisation for Economic Cooperation and Development) sediment (5% peat, 75% sand, 20% kaolinite), which were then placed in larger vessels with the M4 medium. The plants were observed for main and secondary shoot length, biomass and root formation within 2–3 weeks of planting. Growth rates were calculated for total plant length and biomass. The variance between the replicates was low throughout the experiment [coefficient of variation (CV) < 26% for total plant length, and between 16 and 40% for biomass]. Relative growth rates based on total plant length were determined as 0.028 and 0.050 per day for M. spicatum in the systems containing M4 medium only and medium plus sediment respectively. Similar results were observed for E. canadensis, with relative growth rates of 0.26 and 0.073 per day in the two test systems. The root‐shoot ratio at harvest was greater by a factor of 2–3 for E. canadensis in the M4 medium than in the system containing sediment. However, comparable ratios were observed for M. spicatum in the two test systems. Both growth in total plant length and growth in biomass of the two species have potential as measures of toxicity. Copyright © 2006 Society of Chemical Industry