Water samples were taken in Hidden Lake at five different time points around two rotenone applications: (i) five weeks prior to the first rotenone application, on July 12 2018; (ii) approximately three weeks after the first application of rotenone, on 7 September 2018; (iii) approximately 10 months after the first rotenone application, on 10 July 2019; (iv) four weeks following the final treatment of rotenone on the 17 September 2019; and (v) one year after the final rotenone treatment, on 19 August 2020. For each time point there is a pelagic, a littoral and a profundal sample. COI and 18S metabarcoding methods were used to produce community data. The objective of this study was to assess the non-target effect of rotenone application (in summer 2018) on aquatic communities (i.e. phytoplankton, fungi, zooplankton and benthic macroinvertebrates).