This study was conducted in the Talladega research watershed (outlet location: 33.76219799, -85.59550775) in the Talladega National Forest (Cleburne County, AL, USA) as part of the Aquatic Intermittency effects on Microbiomes in Streams (AIMS) project, an NSF EPSCoR funded project (OIA 2019603). The project sought to explore the impacts of stream drying on downstream water quality across Kansas, Oklahoma, Alabama, and Idaho, integrating datasets on hydrology, microbiomes, macroinvertebrates, and biogeochemistry. The Talladega watershed drains a non-perennial unnamed tributary of Pendergrass Creek and contains 0.92 km2 of mixed coniferous and deciduous forest in the Piedmont Upland physiographic section. Located near Anniston, AL, the watershed spans an elevation range from 345 to 456 m above sea level and is a tributary to the Coosa River (within the larger Mobile-Tombigbee basin). The region has a humid subtropical climate, with mean daily January and July air temperatures of 5.3°C and 25.3°C respectively, and mean annual precipitation of 1,400 mm/yr.

We collected samples every ~3 weeks from November 27, 2021, to January 31, 2023, at the outlet of our watershed (TLM01, approach 1). This data resource characterizes non-perennial stream sites and other metadata for macroinvertebrate samples collected as part of the NSF-funded project AIMS (Aquatic Intermittency Effects on Microbiomes in Streams), including Approach 1 (maintenance sampling'), Approach 2 ('seasonal sampling') across 7 distributed sites and Approach 3 (spatiialy intense synoptic across 28 sites). Characterization of sampling sites follow the AIMS Macroinvertebrate Field Sampling SOP (Allen & Busch, 2024). Briefly, 100 m transects were marked, with AIMS STIC centers at 50m. Samples were collected at 0, 20, 40, 60, 80, and 100 m and compiled into a single macroinvertebrate sample. Field data was collected at each transect depending on whether the sampling crew observed surface water in the stream channel at the time of sampling ('WetDry'), a visual estimation of benthic sediment and algae cover percentage, percent canopy cover (estimated via densiometer), the wetted width of the stream at each sampling location where possible, and if the full 100m reach was connected and flowing. Some measurements are missing from sites due to logistical/human errors, and are recorded as NAs. Wetted width could only be collected when surface water was present, so NAs associated with dry sites are flagged ('WW_flag' variable) for convenience. In addition, substrate estimates, canopy cover, habitat classifcation, habitat isolation, and depth were not recorded for dry sites.