Methods

Sampling methods are described by Szydlowski et al. "Macrophyte and snail community responses to 30 years of population declines of invasive rusty crayfish (Faxonius rusticus)," but are provided here for convenience. Instrumentation is further documented in the supplementary information of the paper.<br/>Sampling methods are described by Szydlowski et al. "Macrophyte and snail community responses to 30 years of population declines of invasive rusty crayfish (Faxonius rusticus)," but are provided here for convenience. Instrumentation is further documented in the supplementary information of the paper.<br/>Sampling methods are described by Szydlowski et al. "Macrophyte and snail community responses to 30 years of population declines of invasive rusty crayfish (Faxonius rusticus)," but are provided here for convenience. Instrumentation is further documented in the supplementary information of the paper.<br/>Sampling methods are described by Szydlowski et al. "Macrophyte and snail community responses to 30 years of population declines of invasive rusty crayfish (Faxonius rusticus)," but are provided here for convenience. Instrumentation is further documented in the supplementary information of the paper.<br/>Sampling methods are described by Szydlowski et al. "Macrophyte and snail community responses to 30 years of population declines of invasive rusty crayfish (Faxonius rusticus)," but are provided here for convenience. Instrumentation is further documented in the supplementary information of the paper.<br/>Sampling methods are described by Szydlowski et al. "Macrophyte and snail community responses to 30 years of population declines of invasive rusty crayfish (Faxonius rusticus)," but are provided here for convenience. Instrumentation is further documented in the supplementary information of the paper.<br/>Sampling methods are described by Szydlowski et al. "Macrophyte and snail community responses to 30 years of population declines of invasive rusty crayfish (Faxonius rusticus)," but are provided here for convenience. Instrumentation is further documented in the supplementary information of the paper.<br/>Sampling methods are described by Szydlowski et al. "Macrophyte and snail community responses to 30 years of population declines of invasive rusty crayfish (Faxonius rusticus)," but are provided here for convenience. Instrumentation is further documented in the supplementary information of the paper.<br/>Sampling methods are described by Szydlowski et al. "Macrophyte and snail community responses to 30 years of population declines of invasive rusty crayfish (Faxonius rusticus)," but are provided here for convenience. Instrumentation is further documented in the supplementary information of the paper.<br/>

Methods

Phytoplankton samples were obtained from the epilimnion and hypolimnion by
slowly lowering weighted Tygon tubing through the water column to a depth of 7 m, such
that the tubing was filled with a representative water column sample. Based on the inner
diameter of the tubing, 205 mL of water was pumped from the tubing for the epilimnion
sample. Next, 267 mL of water was pumped from the tubing for the hypolimnion sample. Each
sample was collected into a 250 mL amber bottle that contained 2 mL of Lugol’s solution.
Phytoplankton samples were pooled by sampling date and sent to Phycotech Inc. (St. Joseph
MI, USA) for phytoplankton identification, and concentration and biovolume
quantification.<br/>

Methods

Our study lake, South Sparkling Bog (SSB) (46.003°N, 89.705°W), is a bog lake
located in Vilas County in Northern Wisconsin. South Sparkling Bog is a dystrophic,
dimictic lake with a maximum depth of 8 m, a mean depth of 3.6 m, and a surface area of
0.44 ha. South Sparkling Bog is surrounded by a sphagnum bog mat and has no shoreline
development. During the winters of 2019-2020 and 2020-2021, snow was removed from the
surface of South Sparkling Bog following any snow accumulation event. Removal was
conducted via a snowplow attached to the front of an ARGO all-terrain vehicle and a
snowblower. The winter of 2018-2019 served as a reference year, and snow was not removed
from South Sparkling Bog’s surface. While ice cover persisted, plankton samples were
collected at the deep spot for each lake during on a biweekly-to-monthly basis each
winter. On each sampling date, one integrated zooplankton tow was taken at a depth of 0-7
m using a 56 µm mesh Wisconsin net. All zooplankton samples were collected into glass
sample jars, preserved in 90% ethanol, and saved for laboratory analysis. In the lab,
zooplankton samples were filtered through 53 µm mesh and diluted to a known volume, and
three sub-sample replicates were taken using a 1 mL Hensen-Stempel pipette. Sub-sample
replicates were counted to at least 100 individuals, otherwise the entire sample was
quantified. Sub-sample data was then converted to the known diluted volume and finally
converted to total filtered volume (from the integrated tow sample) to estimate density
(individuals L-1). Zooplankton samples were processed using a Leica M8Z dissecting scope
and Leica imaging software. Replicate subsamples were averaged to estimate total abundance
and density, and average lengths (mm) for each sample taxa were calculated from measures
of the first 30 taxa found within a sample date.<br/>

Methods

One zooplankton sample was collected in June 2015 at the deepest part of each lake, via vertical tow with a Wisconsin net (20cm diameter, 80um mesh). Contents of the net were preserved in the field with cold 95% ethanol. Subsamples of each vertical tow sample were counted for zooplankton species, using enough volume to count at least 300 individuals. A larger volume was then visually scanned to look for presence of additional species not seen in the count volume, until at least 2000 individuals had been seen.

Methods

We collect zooplankton samples at the deepest part of the lake using two different gear types. We take one vertical tow with a Wisconsin Net (80um mesh), and a series of Schindler Patalas (53um mesh) samples spanning the water column. All samples are preserved in cold 95percent EtOH.
After collection we combine subsamples of the individual Schindler Patalas trap samples to create one hypsometrically pooled sample for each lakeordate. The individual depth samples are discarded after pooling except from one August sampling date per year. The Hypsometrically Pooled sample and the Wisconsin Net sample are archived in the UW Zoology museum.
We count zooplankton in one or two subsamples, each representing 1.8L of lake water, of the hypsometrically pooled samples to calculate zooplankton abundance. We count one sample date per month from the open water season, and the February ice cover sample. We identify individuals to genus or species, take length measurements, and count eggs and embryos.
Protocol log: 1981-May1984 -- a 0.5m high, 31L Schindler Patalas trap with 80um mesh net was used. Two Wisconsin Net tows were collected. Preservative was 12percent buffered formalin.
June1984 -- changed to 53um mesh net on Schindler trap.
July1986 -- began using the 2m high, 45L Schindler Patalas trap. Changed WI Net collection to take only one tow.
2001 -- changed zooplankton preservative from 12percent buffered formalin to 95percent EtOH.
The number of sample dates per year counted varies with lake and year, from 5 datesoryear to 17 datesoryear.
1981-1983 -- pooled samples are of several types: Total Pooled (TP) were created using equal volume subsamples of the Schindler samples. Epi, Meta, Hypo pooled used equal volume subsamples from the Schindler samples collected from each of the thermal strata. Strata Pooled used equal volume subsamples from the Epi, Meta, Hypo pooled samples to create an entire lake sample. Hypsometrically Pooled (HP) is our standard, which uses subsample volumes weighted to represent the hypsometry of the lake.

Methods

Fisheries surveys of inland lakes and streams in Wisconsin have been conducted by the Wisconsin Department of Natural Resources (WDNR) professionals and its predecessor the Wisconsin Conservation Department for >70 y. Standard fyke net and boat electrofishing surveys tend to dominate the fisheries surveys and data collected. Most fyke net data on certain species (e.g., Walleye Sander vitreus and Muskellunge Esox masquinongy) originates from annual spring netting surveys following ice-out. These data are used for abundance estimates, mark and recapture surveys for estimating population sizes, and egg-take procedures for the hatcheries. Boat-mounted boom and mini-boom electrofishing surveys became increasingly common in the late 1950s and 1960s. Boat electrofishing surveys have typically been conducted during early summer months (May and June), but some electrofishing survey data are also collected in early spring as part of walleye and muskellunge mark-recapture surveys. Summer fyke netting surveys have been collected more sporadically over time, but were once more commonly used as a panfish survey methodology. Surveys were largely non-standardized. Thus, future users and statistical comparisons utilizing these data should acknowledge the non-standard nature of their collection. More in-depth description of these data can be found in Rypel, A. et al., 2016. Seventy-Year Retrospective on Size-Structure Changes in the Recreational Fisheries of Wisconsin. Fisheries, 41, pp.230-243. Available at: http://afs.tandfonline.com/doi/abs/10.1080/03632415.2016.1160894

Methods

Samples counted prior to 1996 were assigned one taxon name with all taxonomic information. This taxon name was split into distinct columns of genus, species and description for archival as best possible. Samples from 2013-2015 were sent to Phycotech inc. (http://www.phycotech.com/) to be counted.

Methods

Protocol available in methods section of: http://msphere.asm.org/content/2/3/e00169-17
Prior to collection, water temperature and dissolved oxygen concentrations are measured using a YSI 550a. The ranges of the epilimnion and hypolimnion are determined based on the location of the thermocline (where temperature/oxygen is changing the fastest). The two layers are collected separately in 1 meter increments using an integrated water column sampler. Water samples are taken back to the lab, shaken thoroughly, and filtered via peristaltic pump through 0.22 micron filters (Pall Supor). Filters are temporarily stored at -20C after collection and then transferred to -80C after transport on dry ice from Trout Lake Station to UW-Madison. Nutrient samples are collected bi-weekly following standard LTER protocols. DNA is extracted from filters using a FASTDNA SpinKit for Soil with minor modifications. (In cases of low yield or specialized sequencing methods, a phenol-chloroform extraction is used instead). The protocol for sequencing and analysis of data varies by year and by sub-project.

Methods

the protocol employed here is based on:
Hauxwell, J., S. Knight, K. Wagner, A. Mikulyuk, M. Nault, M. Porzky and S. Chase . 2010. Recommended baseline monitoring of aquat ic plants in Wisconsin : sampling design, field and laboratory procedures, data entry and analys is, and applica tions. Wisconsin Department of Natural Resources Bureau of Science Services, PUB-SS-1068 2010. Madison, Wisconsin, USA.

Methods

Experimental designThe experiment was conducted from 16 to 26 June 2008. In the first treatment, oxygen was added to the hypolimnion. In the second, nutrients were added to the epilimnion. The third treatment simulated a mixing event (overturn). There also was a control enclosure with no treatment and sampling of the ambient lake water. Throughout this manuscript, we refer to these as Oxygen, Nutrient, Mix, Control and Ambient.Twelve limnocorrals were constructed as enclosures for the experiment. Each limnocorral was cylindrical and extended vertically from the surface of the lake to the sediment (approximately 4 m). The total volume was approximately 5050 l. Details of limnocorral construction are provided in online Supporting Information.The limnocorrals were deployed on 15 June 2008 to allow the sediment and water column to stabilize before treatment on 16 June 2008. The limnocorrals were deployed in a random spatial arrangement throughout the lake, at a maximum depth of 3.25 to 3.5 m. Replicates from each treatment were instrumented with a chain of HOBO temperature sensors (Onset), and one replicate from each had a self-logging DO sonde (Yellow Springs Incorporated) in the hypolimnion (3 m depth). More thermistors were deployed in the Oxygen and Mix treatments because thermal stratification was important for evaluating success of these treatments.For the Mix treatment, a 60 cm flat disk was raised and lowered between 3.5 m depth and the lake surface. Holes were drilled through the disk surface to increase turbulence (Sanford, 1997; Regel et al., 2004). A brick was tied underneath the disk to maintain stability. We manually oscillated the disk every 10 min for 1 h and then, after a 1 h break, continued for an additional hour. Temperature and DO profiles were monitored within the limnocorral with a hand-held probe to track mixing progress.The goal of the Oxygen treatment was to aerate the hypolimnion water without allowing it to mix with the epilimnion. This treatment was achieved by pumping hypolimnion water from the bottom of the limnocorral into an external cooler where the water was aerated with bubble diffusers, and then returned to the bottom of the limnocorral (Fig. S1). Valves on a compressed air cylinder were used to control the delivery of air to the coolers. One cooler was maintained for each replicate limnocorral. Thermally insulated tubing was used to transport water. A thermistor was deployed in each cooler to ensure ambient hypolimnion temperature was maintained. The water was removed and returned using two linear diffusers that were 0.6 m in length, spanning a depth range of approximately 2.5&ndash;3.1 m within the hypolimnion. The diffusers faced inward with 0.5 m fixed distance between them, retained by a plastic divider. This treatment was applied continuously over 3 h, until DO concentrations increased.The Nutrient treatment was achieved by adding ammonium chloride (NH4Cl) and potassium phosphate monobasic (KH2PO4) as N and P sources. These compounds were chosen because they are commonly bioavailable sources of nutrients. P was added to the epilimnion to achieve a final concentration of 3 micro g P l&minus;1, which was approximately the average concentration expected in the mixed water column. This value was based on nutrient analyses from integrated water collected on 9 June 2008 in North Sparkling Bog, a week prior to experiment start. Similarly, N was added to achieve a final concentration of 70 &micro;g N l&minus;1. The limnocorral s epilimnion volume (0&ndash;2 m integrated depth) was calculated to be 2520 l, and we used the molar mass to determine the amount of each nutrient added to the epilimnion to achieve the expected mixed concentration. Dry chemicals were dissolved into to 500 ml of surface water from each limnocorral, and then added into each separately. Rationale for directly manipulating only one layer in the Oxygen and Nutrient treatments is given in the online Supporting Information.The Control limnocorrals were left undisturbed. To prevent mixing during equipment removal, all tubing was left inside the limnocorrals until the experiment ended.