Understanding the effects of fungicides on non-target organisms at realistic concentrations and exposure durations is vital for determining potential impacts on aquatic ecosystems. Environmental concentrations of the fungicide azoxystrobin have been reported up to 4.6µg/L in the United States and 30 µg/L in Europe. The objective of the present study was to evaluate the chronic toxicity of azoxystrobin in water-only exposures with an amphipod (Hyalella azteca; 42-d exposure), a midge (Chironomus dilutus; 50-d exposure), a cladoceran (Ceriodaphnia dubia; 7-d exposure), and a unionid mussel (Lampsilis siliquoidea; 28-d exposure) at environmentally relevant concentrations. The potential photo-enhanced toxicity of azoxystrobin accumulated by C. dubia and L. siliquoidea following chronic exposures to azoxystrobin was also evaluated. The 20% effect concentrations (EC20s) based on the most sensitive endpoint were 4.2 µg/L for H. azteca reproduction, 12 µg/L for C. dubia reproduction and C. dilutus emergence, and >28 µg/L for L. siliquoidea. Hyalella azteca was more sensitive to azoxystrobin compared to the other three species in the chronic exposures. There was no photo-enhanced toxicity observed for either C. dubia or L. siliquoidea exposed to ultraviolet light in control water following azoxystrobin tests. Results of the present study indicate chronic effects of azoxystrobin on 3 of 4 invertebrates tested at environmentally relevant concentrations. The changes noted in biomass and reproduction have the potential to alter the rate of ecological processes driven by aquatic invertebrates. This article is protected by copyright. All rights reserved.