These data were collected in support of the sampling goals of the Aquatic Intermittency effects on Microbiomes in Streams (AIMS) Project. We sampled macroinvertebrates along a 100 m reach every 20 m alternating channel location from left, to center, to right for a total of six samples per reach. We compiled all six samples into a composite sample, removed debris and leaf matter via manual inspection of material and elutriation, and sieved the remaining sample through a 500 µm sieve. We then preserved the sample in 95% ethanol and returned it to the lab. In the lab, ethanol was refreshed before we froze the sample in a -20 ºC freezer. We shipped samples to a commercial lab (Jonah Ventures, Boulder, CO) for metabarcoding analysis of the invertebrate community. Briefly, Jonah Ventures homogenized community samples with a hand immersion blender and extracted DNA with a DNeasy Powersoil Kit following the manufacturer’s protocol. They then amplified samples using two primers from the CO1 gene (CO1 F230 fragment Hajibabaei et al. 2012; CO1 BE fragment Folmer et al. 1994, Gibson et al. 2015). They conducted PCRs with initial denaturation at 95 ºC for 5 min, followed by 40 cycles of 40s at 95 ºC, 1 min at 46 ºC, 30s at 72 ºC and a final elongation at 72 ºC for 10 min. They then cleaned samples using Exo1/SAP, and pooled, normalized and indexed. Samples were sent for sequencing at the CU Boulder BioFrontiers Sequencing Center where the Center used the v2 500-cycle kit with appropriate quality control measures. Jonah Ventures then demuliplexed sequenced samples using phigs v2.1.0 followed by removal of gene primers and merging read pairs. Read pairs were then clustered using unoise3 denoising algorithm in vsearch and with sequences with less than 8 reads discarded. Taxonomy was assigned using a custom best-hits algorithm with reference to NCBI Gen Bank to each Exact Sequence Variant (ESV) (See linked Jonah Venture documents for full methods).

KNZ - This study was conducted on the South Fork of the Kings Creek research watershed (outlet location: 39.092281, -96.58719) within Konza Prairie Biological Station (KBPS) near Manhattan (KS, USA). At the USGS gage located on the mainstem (06879560; est. 1979), Kings Creek is a 5th order intermittent stream draining 1059-ha of tallgrass prairie that is actively managed using controlled buns at varying frequencies (1-20 year return intervals) and grazing by bison or cattle. Kings Creek ranges in elevation from 338 to 430 m above sea level and drains into the Kansas River. The region is within a midwestern continental climate, with temperatures ranging from 4 to 22°C and mean annual precipitation averaging about 780 mm/yr.

This site lies within the Flint Hills ecoregion of eastern KS and northeastern OK, has a mean annual temperature of 11.7ºC (1983-2020), and 811 mm annual precipitation (1983-2020) with high interannual variability and an estimated 75% of annual precipitation occurring during late spring and early summer (Hayden 1998, Sadayappan et al. 2023). The AIMS study catchment, the South Fork of Kings Creek, is grazed by bison year-round, and includes sub-catchments with variable prescribed burn history, but the entire study area was burned in early April 2021. The riparian vegetation is deciduous gallery forest and the highest portion of the landscape is tallgrass prairie (Dodds et al. 2004) dominated by warm-season grasses, though woody encroachment has occurred in most subwatersheds (Sadayappan et al. 2023)*. The underlying bedrock of the Flint Hills ecotone is characterized as limestone, mudstone, and shale with predominately silty clay loam soils that rest atop (Hayden 1998, Vero et al. 2018).

Dodds, W. K., et al. 2004. BioScience 54(3):205-216.
Hayden, B. P. 1998. Grassland dynamics: Long-term ecological research in tallgrass prairie. Oxford University Press, New York.
Vero, S., G. et al. 2018. Vadose Zone Journal 17(1):1-11.

YMR - Youngmeyer Ranch is located in Elk County, KS (outlet location: 37.56442, -96.49106) and managed by Witchita State University and owned by the Youngmeyer Trust. The ranch is roughly 1902 ha of grassland prairie used predominantly for cattle grazing and is burned every 1-2 years. A tributary of the Elk River, specifically the south branch of the Elk River headwaters, elevation at Youngmeyer ranges from 373-488 m with mean annual temperature of 13.7°C and mean annual precipitation of 979mm. This site is geologically constructed of Permian age limestone and shale with layers of chert below silty clay loam soils (Houseman et al. 2016). This site is predominantly grassland composed of the same dominant grasses as Konza Prairie, with scattered black oaks (Q. veluntina) along the creeks (Houseman et al. 2016).

Houseman, G. R., M. S. Kraushar, and C. M. Rogers. 2016. Transactions of the Kansas Academy of Science 119(1):27-32.