Abstract: In North America, Eurasian watermilfoil Myriophyllum spicatum is one of the most widespread non-native aquatic plant species in freshwater ecosystems.Applications of 2,4-dichlorophenoxyacetic acid (2,4-D) herbicides are often utilized to control Eurasian watermilfoil.Herbicide applications may have unintended effects on non-target organisms like zooplankton and fish, but these effects are poorly understood.Our objectives were to determine if whole-lake 2,4-D herbicide treatments (epilimnetic target rate = 0.3 ppm) used to control Eurasian watermilfoil affected (1) d., diversity, and size of zooplankton and (2) d., diversity, and growth of larval fishes in three northern Wisconsin lakes when compared to trends observed in three untreated reference lakes.Average peak concentrations of 2,4-D following applications ranged from 0.152 to 0.257 ppm.We found no statistically significant evidence that the applications influenced the zooplankton and larval fish metrics we examined, but subtle responses to the herbicides may not have been detected given the inherent variation in our response metrics.Addnl. replication and research are needed to assess responses to higher concentrations and repeated applications over time and to document responses across a broader range of lake types and fish communities.