DOI: 10.5061/dryad.ncjsxkszk
title: Data and code for: Facilitating the recovery of insect communities in restored streams by increasing oviposition habitat

description: Recruitment limitation is known to influence species abundances and
 distributions. Recognition of how and why it occurs both in natural and in
 designed environments could improve restoration. Aquatic insects, for
 instance, rarely re-establish in restored streams to levels comparable to
 reference streams even years after post-restoration. We experimentally
 increased oviposition habitat in five out of ten restored streams in
 western North Carolina to test whether insect egg-laying habitat was
 limiting insect populations in restored streams. A main goal was to test
 whether adding oviposition habitat in the form of rocks that partially
 protrude above the water surface could be used to increase the abundance
 and richness of stream insect eggs and larval insects in restored streams.
 Adding egg-laying habitat enhanced several response variables (e.g.,
 protruding rocks, number of eggs, egg masses, egg morphotype richness, and
 oviposition habitat stability) to levels similar to those found in
 reference streams. Following the addition of protruding rocks, egg mass
 abundance increased by 186 % and richness increased by 77 % respectively
 in restored-treated streams. Densities of larval insects that attached
 their eggs to protruding rocks showed an overall pattern consistent with
 treatment effects due to the combination of non-significant and
 significant increases of several taxa and not just one taxon. Our results
 indicate that these stream insect populations are limited by oviposition
 habitat and that adding egg-laying habitat alleviated this component of
 recruitment limitation. However, the weaker larval response indicates that
 additional post-recruitment factors, such as egg or larval mortality, may
 still be limiting a full recovery of larval insect abundances in these
 restored streams. This study shows the importance of integrating
 information from animal life histories, ecology, and geomorphology into
 restoration practices to improve the recovery of aquatic insects, which
 are commonly used to assess water quality and the biological efficacy of
 stream restoration.
description: Site selection We obtained a list of 378
 North Carolina Department of Environmental Quality projects that included
 a stream component. To reduce variability, we selected streams with
 similar restoration priorities, land-use, and stream bed composition. We
 selected projects that utilized restoration priority type one or two (RP1
 or RP2) of the four Rosgen (1997) developed options for the restoration of
 incised channels. These project types increased channel sinuosity, stream
 bed heterogeneity, and either elevated channels to reconnect them to
 floodplains (RP1) or created floodplains at the current stream elevation
 (RP2). We also selected projects 7–15 years post-restoration to allow
 sufficient time for riparian planting to mature, fine sediments to
 stabilize, detritus to accumulate, biofilms to establish, and some insects
 to recolonize. Further, we selected streams with similar catchment
 land-use composition (e.g., impervious surface less than 5 % and developed
 land less than 20 %). We identified large cobble- and boulder-sized rocks
 (128–256 and &gt;256 mm) as the oviposition habitat for manipulation.
 Consequently, we excluded streams located in the coastal plain and with a
 median particle size (D50) &lt; 2 mm (i.e., primarily sand-bed
 streams) because, given their hydrogeomorphic context, these streams often
 naturally lack this type of oviposition habitat and we wanted to keep
 oviposition habitat type similar across streams. Although stream insects
 use rocks, wood, and live vegetation as well as other materials for
 egg-laying (Smith and Storey 2018), we focused on rocks in this study
 because they were the most abundant habitat, more permanent than organic
 materials such as wood, and are usually available or added during stream
 restoration. Ultimately, 12 restoration projects fit
 all requirements including access. Following physical visits, we selected
 10 restored streams. The 10 streams were part of four restoration projects
 located in western North Carolina (Appendix S1: Figure S1). We grouped
 streams primarily by proximity to one another (Appendix S1: Table S1;
 Figure S1) and secondarily by stream characteristics, such as channel size
 and presence or absence of a culvert above the restored reach (Appendix
 S1: Table S1). We randomly assigned one stream from each of five groups to
 receive the treatment (addition of suitable rocks for oviposition) using a
 coin toss, resulting in five restored-untreated and five restored-treated
 streams (Appendix S1: Table S1). Two projects included multiple streams
 (at least one treated and untreated). For example, the two streams at
 Warrior Creek were located on different tributaries. Morgan Creek had four
 streams all located on different tributaries and the main channel included
 an upstream untreated and downstream treated section of stream. We were
 limited to three reference streams because of a lack of reference streams
 with comparable catchment size, forested watershed in reasonable proximity
 to the restored streams and limited access to private property (Appendix
 S1: Table S1; Figure S1). Reference streams were on large areas of state
 or federal government land that have provided watershed-scale protection
 from the land use changes affecting restored streams (e.g., cattle grazing
 and forest clearing for pasture and crops). The Indian Creek reference
 site was ~10 m upstream of a State of North Carolina Division of
 Environmental Quality (NCDEQ) reference site near Hanging Rock State Park
 and was near both Pinch Gut and Candiff Creek. Boone Fork Creek was
 nearest to Warrior Creek but at a higher elevation in the northeastern
 portion of Pisgah National Forest, Lenoir, NC. Beaverdam Creek was located
 east of Morgan Creek on the southern end of the South Mountains State Game
 Lands. One reference stream was included in each of groups four and five
 with restored-untreated and restored-treated streams, but the reference
 stream Beaverdam Creek was included in its own group, six (Appendix S1:
 Table S1). Groups 1–3 did not include a reference stream, resulting in a
 partially balanced incomplete block design (see Statistical
 Analysis).  Experimental addition of oviposition
 habitats To experimentally test whether adding
 protruding rocks, or potentially suitable oviposition habitat, increased
 the abundance and richness of insect egg masses and larval abundance in
 restored streams, we used a before and after repeated measures ANOVA
 incomplete block experimental design with three treatment types: five
 replicate restored streams in which we manipulated oviposition habitats by
 adding suitable rocks (restored-treated), five replicate restored streams
 that were unmanipulated (restored-untreated), and three reference streams
 (reference). We planned to treat the five restored streams in March 2020
 before adult insect emergence, and census egg inputs from May to October,
 similar to 2019. However, due to the COVID-19 pandemic, treatment was
 delayed until May 2020 and egg mass censuses started in June 2020
 (Appendix S1: Fig S2). Because of this delay and the seasonality of
 oviposition that includes a May peak, we omitted 2019 May data from the
 analyses and used egg censuses from June – October for both 2019 (before)
 and 2020 (after). In May 2020, we added or repositioned
 rocks within the five restored-treated streams (Table 1; Appendix S1:
 Figure S3) to increase the availability of suitable oviposition habitat.
 Two people added rocks by hand to all five 50-meter-long sections of
 restored-treated streams in a single visit (Table 1). Rocks added were
 collected from nearby riparian areas or were formerly submerged within the
 channel; hence, protruding rocks were weathered and included rounded
 native stone but most were angular quarry stones added during restoration.
 Rock size (measured as the intermediate axis width) varied between 181–512
 mm depending on availability within or near treatment streams and whether
 they were large enough to protrude above the water surface. We positioned
 rocks, with 25–30 % of the surface protruding above the water, in deep
 portions of riffles or other areas with fast to moderate water velocity
 (e.g., runs) where fine sediment deposition was low and thus the
 likelihood of rocks becoming embedded by sediment was low. Rocks were not
 placed in locations with low water velocity and high fine sediment
 deposition, such as pools, or low water velocity and shallow areas along
 the streambank where attached eggs are more likely to dry as water levels
 recede. We adjusted rocks throughout the study period (June-October 2020)
 to ensure the supplemental habitats remained suitable (i.e., protruding
 and unembedded) for egg-laying females. Our rationale for repositioning
 potential egg-laying habitat was to ensure the treatment was maintained.
 This decision reflects our aim of investigating the outcomes of maximizing
 availability of potential oviposition habitat to provide a “proof of
 concept” test of our hypotheses. The availability of these habitats is a
 dynamic naturally occurring process in streams (Ciotti et al. 2021), but
 this process may be delayed or impaired in many restored streams, as
 evidenced by the lack of egg-laying habitat in these streams (Jordt and
 Taylor 2021a). We used the mean number of suitable rocks as the response
 variable for protruding rock abundance, which was calculated as the sum of
 the number of suitable rocks per m2 stream area for each stream across all
 censuses for each year (omitting May 2019 census data), divided by the
 number of censuses for each year (2019 and 2020).
 Protruding rocks absent on subsequent sampling dates were
 recorded as missing (i.e., completely buried or rolled out of the study
 area). Missing rocks were removed from the count of protruding rocks
 present but were included in counts if they were rediscovered on
 subsequent sampling dates. We noted whether rocks rolled (as evidence by
 the position of the painted number), were underwater, out of water and
 dry, and/or embedded in sediments. With the exception of treatment streams
 (during 2020), we did not unembed rocks to census them. In treatment
 streams in 2020, rocks that became unsuitable (i.e., they became embedded
 or submerged) were repositioned or unembedded throughout the 2020
 egg-laying season in order to maintain a high level of suitable rock
 availability. Suitable rocks that became embedded or submerged were not
 considered as unstable, or included in the analysis of rock movement.
 Lastly, in restored-treated streams, rocks that were within the study area
 and became suitable for egg laying (became unembedded and protruding) or
 rolled into the study area were repositioned and counted as protruding
 rocks during the 2020 census. Rocks that became suitable (i.e.,
 protruding) because water depth declined or because they rolled into the
 study area were not considered unstable unless subsequently buried,
 rolled, or missing.  Egg mass and egg abundance, egg
 mass morphotype richness, and aggregation In 2019, the
 year prior to adding rocks to streams, we censused a 50 m length of the
 ten restored and three reference streams for egg masses throughout the
 egg-laying period (May to October). Suitable egg-laying habitats were
 identified as protruding rocks with submerged spaces that adult insects
 could access along the sides and underneath (unembedded), as several
 previous studies have shown these specific rocks to be important stream
 insect egg-laying habitat (Peckarsky et al. 2000, Reich and Downes 2003b,
 Encalada and Peckarsky 2006, 2007, Miller et al. 2020). Protruding rocks
 were numbered using a nontoxic paint pen and removed from the water to
 identify egg masses to family or morphotype, count egg masses, photograph,
 and make sketches of mass type and locations on each rock. New masses on
 rocks were identified by comparing sketches to those made previously.
 Rocks were placed back into the stream in their original location and
 position. Individual egg masses were distinguished from one another by the
 gelatinous matrix, or spumaline surrounding each egg mass, and by size,
 shape, color, and spatial ordering of eggs within each mass. We returned
 every one to two weeks to census existing and newly protruding rocks for
 egg masses 10 times each in 2019 and 2020. We were not able to quantify or
 identify the eggs of some insects that lay individual eggs, egg masses in
 rock crevasses, or in moss growing on rocks (e.g., some Trichoptera,
 Tipulidae, and Elmidae). Egg masses were identified to
 order using published descriptions and photographs (Murvosh 1971, Williams
 1982, Nolte 1993, Peckarsky et al. 2000, Hoffmann and Resh 2003, Reich
 2004, Reich and Downes 2004, Lancaster et al. 2010a, Lancaster et al.
 2010b, Lancaster and Downes 2014, Smith and Storey 2018, Miller et al.
 2020, Jordt and Taylor 2021a) or by identifying females actively
 ovipositing. Although family-level identification of eggs was possible for
 some taxa (e.g., Baetidae, Psephenidae), we could not identify the
 majority of egg masses beyond order, however, we were able to identify
 distinct morphotypes within orders based on color, shape (e.g., mat,
 globular, dendritic, strings, tombstone), texture (e.g., smooth, rough,
 firm) and relative size of the mass (e.g., globular tiny, globular
 strings, mat white, mat yellow). Distinguishing egg masses by morphotype
 provides finer taxonomic resolution when testing for changes in richness
 in restored-treated streams. As the response variable
 for egg inputs, we used the mean number of egg masses (either total masses
 or by morphotype) per stream area, which was calculated as the sum of the
 number of egg masses divided by the stream area censused for each stream,
 divided by the number of censuses for each year (2019 and 2020). Egg
 masses per stream area censused was used rather than total stream area of
 each 50 m study section because, on some occasions (8 of 222), the entire
 50 m study section could not be censused due to lightning storms. We also
 calculated the mean number of egg masses per individual rock area, as
 previous research revealed when rocks were scarce the density of egg
 masses on individual rocks is high (Jordt and Taylor 2021b) and more
 aggregated (Lancaster et al. 2021); therefore, we tested whether the
 density of egg masses on individual rocks in the restored-treated streams
 changed from before to after treatment. Mean egg masses per individual
 rock area was calculated as the mean of the sum of masses on an individual
 rock divided by the underside two-dimensional area of the rock for each
 stream, census date, and year. Larval insect
 abundance We sampled the abundance of benthic larval
 invertebrates during May 2020 (before rock addition) and March 2021 (9
 months after adding rocks) using a 0.09 m<sup>2</sup> Surber
 sampler with 363 µm mesh size net. We selected springtime for larval
 sampling because by this time all eggs oviposited the previous year should
 have hatched (Appendix S1: Figure S2), and also because we were primarily
 interested in whether the additional egg-laying habitat caused an increase
 in larvae that survived until the next egg-laying period and could
 potentially emerge into egg-laying adults. Hence, we were not interested
 in quantifying transient increases in larval abundance. Invertebrates were
 preserved in ethanol, stained with Rose Bengal stain, picked under 10X
 magnification, and counted and identified to order and family because this
 was the lowest taxonomic resolution we could identify eggs, as there are
 currently no taxonomic keys for eggs. Statistical
 analysis We used a linear mixed-model,
 repeated-measures ANOVA (proc mixed in SAS) partially balanced incomplete
 block design to analyze whether adding rocks increased 1) the abundance of
 suitable oviposition habitat, 2) the density of eggs, and density and
 richness of egg masses, 3) the density of larval insects, and 4) the
 stability of suitable oviposition habitat (i.e., decreased the percent of
 rocks rolled, buried, or missing). This was a partially balanced
 incomplete block design because references did not occur in all blocks;
 therefore, the precision of differences in restored-treated versus
 restored-untreated was greater than the precision of differences in
 restored-treated versus reference streams (Oehlert 2000). We used
 pre-planned comparisons, or a priori contrasts, because we were interested
 in testing specific and sometimes directional hypotheses associated with
 the stream type x time interaction. For example, a before-after increase
 of a response in restored-treated streams compared to restored-untreated
 streams. For the number of rocks rolled, buried, or missing, we
 hypothesized a decrease in restored-treated streams compared to
 restored-untreated streams and reference streams. For all tests, including
 a priori contrasts, we report two-tailed test p values. We omitted a
 priori contrast between restored-untreated and reference streams because
 this was already tested by Jordt and Taylor (2021b). For unplanned
 comparisons, such as comparing the means of restored-treated streams
 versus reference streams after adding rocks, we reported Tukey-Kramer
 two-tailed test statistics only if the type III ANOVA main effects p value
 was &lt; 0.05, an adjustment to reduce type I error. Stream type
 (i.e., restored-untreated, restored-treated, and reference), time (2019
 and 2020), and their interaction were fixed factors. Group (i.e., block)
 and group x stream type were random factors. For larval density analyses,
 we fit generalized linear mixed models (proc glimmix in SAS) with the same
 fixed and random effects as for linear mixed models above and assessed the
 fit of either gamma or lognormal distributions using Akaike’s information
 criterion (AIC). We did not adjust for multiple tests in the analyses of
 changes in multiple egg and larval taxonomic groups because in some cases
 the model structure did not support it and such adjustments with limited
 replicates (n=5) increase the likelihood of type II errors. Instead, we
 report unadjusted probabilities and the magnitude of effects and do not
 use any one significant result to make broad overall conclusions (Voelkl
 2019). We used linear regression to test whether annual egg mass densities
 per stream area of all taxa combined increased as the stability of
 protruding rocks (i.e., percentage of suitable rocks rolled or missing)
 decreased in each stream, regardless of type. We also used linear
 regression to assess the relation between changes in protruding rock
 densities and changes in egg mass density and egg mass morphotype richness
 from 2019 to 2020, as this could be used to predict the effects of the
 number of rocks added on egg masses density and richness. We tested
 relationships between rock size and cumulative number of egg masses per
 rock per year using a negative binomial regression and the relationship
 between rock size and probability of egg masses per rock per year using
 logistic regression. We transformed response variables to meet assumptions
 of normality and homoscedasticity. For graphical display of transformed
 variables, we report the back-transformed means (an estimate similar to
 the median in the untransformed scale) and back-transformed 95 %
 confidence intervals. Egg masses were not distributed
 randomly among rocks, so the mean density of egg masses per protruding
 rock area was not a suitable measure of aggregation or crowding. The
 distribution of number of egg masses per protruding rock and the number
 per individual rock area were highly right-skewed. Therefore, we analyzed
 the mean number of egg masses per individual rock area for rocks with
 &gt; 0 egg masses. Additionally, to assess aggregation, or crowding,
 of egg masses on individual protruding rocks among streams and whether
 aggregation changed following the addition of protruding rocks, we used a
 density-independent metric developed by Lang et al. (2017, eq. 23 with
 q=1), which is more appropriate than Lloyd’s (1967) patchiness index when
 mean abundances are small. As a descriptive measure of the distribution of
 egg masses on individual rocks, we calculated egg mass prevalence as the
 number of protruding rocks with one or more egg masses divided by the
 number of all protruding rocks, including those with zero egg masses. All
 statistical analyses were performed using SAS OnDemand for Academics (SAS
 Institute Inc. 2014).
description: This README file was generated on 2023-10-31 by Brad Taylor. GENERAL
 INFORMATION 1. Title of Dataset: Data and code for: Facilitating the
 recovery of insect communities in restored streams by increasing
 oviposition habitat. 2. Author Information A. Principal Investigator
 Contact Information Name: Brad Taylor Institution: North Carolina State
 University Address: Raleigh, NC USA Email: bwtaylo3@ncsu.edu B. Associate
 or Co-investigator Contact Information Name: Samantha Dilworth (formerly
 Jordt) Institution: University of Wyoming Address: Laramie, WY USA Email:
 samanthajordt@gmail.com 4. Date of data collection (single date, range,
 approximate date): 2019-2020 5. Geographic location of data collection:
 western, North Carolina 6. Information about funding sources that
 supported the collection of the data: North Carolina Department of
 Environmental Quality SHARING/ACCESS INFORMATION 1. Licenses/restrictions
 placed on the data: CC0 1.0 Universal (CC0 1.0) Public Domain 2. Links to
 publications that cite or use the data: Dilworth, S. and Taylor, B.W.
 (2023). Facilitating the recovery of insect communities in restored
 streams by increasing oviposition habitat. Ecological Applications. 3.
 Links to other publicly accessible locations of the data: None 4.
 Links/relationships to ancillary data sets: None 5. Was data derived from
 another source? No A. If yes, list source(s): NA 6. Recommended citation
 for this dataset: Dilworth, S. and Taylor, B.W. (2023). Facilitating the
 recovery of insect communities in restored streams by increasing
 oviposition habitat. https://doi.org/10.5061/dryad.ncjsxkszk DATA
 & FILE OVERVIEW 1. File List: A) site\_data.csv B) site\_metadata
 C) site\_visit\_data.csv D) site\_visit\_metadata.csv E)
 EggMassQuantities\_2020\_data.csv F)
 EggMassQuantities\_2020\_data\_metadata.csv G) EggMassData\_2019\_2020.csv
 H) EggMassData\_2019\_2020\_Metadata.csv I) rolled\_rocks.csv J)
 rolled\_rocks\_metadata.csv K) Larvae\_Family.csv L)
 Larvae\_Family\_metadata.csv M) SJordt\_main\_script\_15June2022.R N)
 output\_files\_from\_R\_code.zip O) sas\_code\_files.zip P)
 TableS1-S3-FigureS2-S4-S5-S6-S7-S8.zip Q) Appendix\_S1.pdf R) readme.md
 (this file) 2. Relationship between files, if important: R and SAS code
 read or write the csv files as indicated below. 3. Additional related data
 collected that was not included in the current data package: None 4. Are
 there multiple versions of the dataset? No A. If yes, name of file(s) that
 was updated: NA i. Why was the file updated? NA ii. When was the file
 updated? NA GENERAL OVERVIEW AND DESCRIPTION OF THE STUDY The data and
 code are for an experiment testing weather rocks protruding above the
 stream water limit egg and larval recruitment in previously restored
 streams. We surveyed the different types and abundances of stream insect
 egg masses and larvae in restored and reference streams before and after
 adding protruding rocks (egg-laying habitat for stream insects) to a
 subset of the restored streams, which we refer to as restored-treated. Ten
 restored streams were sampled before and after protruding rocks were added
 to half (n=5) of the restored streams. Three reference streams that have
 not needed restoration were sampled before and after to assess inter
 annual variation as well as for comparison to restored-untreated and
 restored-treated. The before data represent ambient abundances and
 taxonomic types of stream insect egg masses and larvae. The after data in
 restored-treated streams represent the effect of the treatment, the
 addition of egg-laying habitat, or protruding rocks.
 #########################################################################
 DATA-SPECIFIC INFORMATION FOR: site\_data.csv 1. Number of variables: 36
 2. Number of cases/rows: 18 3. Variable List: see site\_metadata.csv 4.
 Missing data codes: NA 5. Specialized formats or other abbreviations used:
 see site\_metadata.csv
 #########################################################################
 DATA-SPECIFIC INFORMATION FOR: site\_visit\_data.csv 1. Number of
 variables: 9 2. Number of cases/rows: 255 3. Variable List:
 site\_visit\_metadata.csv 4. Missing data codes: NA (data not available)
 5. Specialized formats or other abbreviations used:
 site\_visit\_metadata.csv
 #########################################################################
 DATA-SPECIFIC INFORMATION FOR: EggMassQuantities\_2020\_data.csv 1. Number
 of variables: 42 2. Number of cases/rows: 7892 3. Variable List: see
 EggMassQuantities\_2020\_data\_metadata.csv 4. Missing data codes: NA
 (data not available) 5. Specialized formats or other abbreviations used:
 see EggMassQuantities\_2020\_data\_metadata.csv
 #########################################################################
 DATA-SPECIFIC INFORMATION FOR: EggMassData\_2019\_2020.csv 1. Number of
 variables: 14 2. Number of cases/rows: 93 3. Variable List: see
 EggMassData\_2019\_2020\_Metadata.csv 4. Missing data codes: NA (data not
 available) 5. Specialized formats or other abbreviations used: see
 EggMassData\_2019\_2020\_Metadata.csv
 #########################################################################
 DATA-SPECIFIC INFORMATION FOR: rolled\_rocks.csv This file contains data
 on whether a rock used for egg-laying rolled and the number and type of
 eggsmasses present before and after rolling. the number of rocks 1. Number
 of variables: 27 2. Number of cases/rows: 173 3. Variable List: see
 rolled\_rocks\_Metadata.csv 4. Missing data codes: NA (data not available)
 5. Specialized formats or other abbreviations used: see
 rolled\_rocks\_Metadata.csv
 #########################################################################
 DATA-SPECIFIC INFORMATION FOR: Larvae\_Family.csv This file contains data
 on larval insect abundance (number per sample). SAS code
 Larval\_FAMILY\_densities\_CODE.sas calculates the densities per m2 based
 on a Hess sampler with an area of 0.09 m2, that is, number of
 individuals/0.09 m2. 1. Number of variables: 67 2. Number of cases/rows:
 131 3. Variable List: see larvae\_Family\_Metadata.csv 4. Missing data
 codes: NA (data not available) 5. Specialized formats or other
 abbreviations used: see larvae\_Family\_Metadata.csv
 #########################################################################
 CODE-SPECIFIC INFORMATION FOR: SJordt\_main\_script\_15June2022.R This is
 the R cade used to organize and process the raw data and calculate means
 by subsite, site, and stream type for each year. This code also creates
 output files that were further analyzed with SAS code files. 1. Data files
 used by this code: EggMassQuantities\_2020\_data.csv
 EggMassData\_2019\_2020.csv rolled\_rocks.csv 2. N=10 output files
 generated by this code: A) thetable2020.csv B) thetableXstreamarea.csv C)
 tableXtime.csv D) rollXtime.csv E) sitesummarytable.csv F) tablextype.csv
 G) eggdensityperrock.csv H) datajoined.csv I) massperindrockm2.csv J)
 Massesperweek.csv 3. Missing data codes: NA (data not available)
 #########################################################################
 DATA-SPECIFIC INFORMATION FOR: output\_files\_from\_R\_code.zip This
 compressed file contains the output (write) files from the code
 SJordt\_main\_script\_15June2022.R 1. File List (N=10 files): A)
 thetable2020.csv B) thetableXstreamarea.csv C) tableXtime.csv D)
 rollXtime.csv E) sitesummarytable.csv F) tablextype.csv G)
 eggdensityperrock.csv H) datajoined.csv I) massperindrockm2.csv J)
 Massesperweek.csv
 #########################################################################
 CODE-SPECIFIC INFORMATION FOR: sas\_code\_files.zip This file contains the
 SAS code used for data processing and statistical analyses. 1. File List
 (N=8 SAS code files): A) Count\_crowding\_index\_CODE.sas Calculates and
 analyzes crowding metrics of eggs based on counts, not per area of rock.
 B) Density\_crowding\_index\_CODE.sas Calculates and analyzes crowding
 metrics of eggs based on density, that is, number of eggs per area of
 rock. C) Larval\_family\_densities\_CODE.sas Calculates and analyzes
 larval insect densities. D) Number\_egg\_mass\_per\_rock\_area\_CODE.sas
 Calculates and analyzes egg masses per unit area of rock. E)
 Number\_of\_egg\_masses\_CODE.sas Calculates and analyzes egg masses per
 unit area of rock. F) Number\_suitable\_rock\_size\_CODE.sas Analyzes the
 two dimensional size of suitable rocks before and after rock additions to
 treated streams. G) Number\_suitable\_rocks\_CODE.sas Analyzes the number
 of suitable rocks before and after rock additions to treated streams. H)
 Rocks\_rolled\_CODE.sas Analyzes data for differences in number of
 suitable rocks rolled or missing before and after rock additions to
 treated streams. 2. Relationship between files, if important: None
 #########################################################################
 DATA-SPECIFIC INFORMATION FOR: Appendix\_S1.pdf This file contains
 supporting information. 1. Missing data codes: NA (not applicable) 2.
 Specialized formats or other abbreviations used: None
 #########################################################################
 DATA-SPECIFIC INFORMATION FOR: TableS1-S3-FigureS2-S4-S5-S6-S7-S8.zip This
 compressed file contains the dat tables and figures from the supporting
 information file Appendix\_S1.pdf.Tables S2, S4, S5 are tables with
 statistics and are not included here. 1. File list (N=10 files)
 Table\_S1.csv Table\_S3.csv FigureS1.jpg FigureS2\_hydrographs.jpg
 FigureS3.jpg FigureS4\_Supplement\_density\_aggregation\_prevalence.jpg
 FigureS5\_Plecoptera\_larval\_densities.jpg
 FigureS6\_All\_insect\_densities.jpg
 FigureS7\_Non-insect\_larval\_densities.jpg
 FigureS8\_Backtransformed\_Regression\_total\_egg\_masses\_vs\_roll\_buried\_or\_missing.jpg 1. Figures S1, S2, S3, S4, S5, S6, S7, S8. Figures S1, S3 are a map and photograph, respectively, all other figures are graphs. Tables S1, S3 are data tables. 2. Missing data codes: NA (not applicable) 3. Specialized formats or other abbreviations used: figures, photographs, and maps are .jpg formatted files. Tables are .csv formatted files. #########################################################################