Methods

We sampled non-zebra mussel benthic macroinvertebrates twice a summer (early June and late August) from 2016-2018 at five depths (1, 3, 5, 8, and 10m) along each of three transects (A-C) running perpendicular to the shore of Lake Mendota. We collected triplicate samples from each site using a 0.625 m-2 circular quadrat and an airlift method with a modified SCUBA tank suction device called an AquaVac. Air was released through a PVC pipe, creating backpressure to lift sediment, which was captured in a 500μm mesh bag and transported in a resealable plastic bag. We chose an airlift method because of difficulty closing Eckman samplers on the hard substrates of rock and zebra mussel druses. Occasionally additional samples were taken with an Eckman, often at deeper depths, for comparing to the main transects and depths sampled with AquaVac or to collect additional material for isotope analysis.<br/>We sampled non-zebra mussel benthic macroinvertebrates twice a summer (early June and late August) from 2016-2018 at five depths (1, 3, 5, 8, and 10m) along each of three transects (A-C) running perpendicular to the shore of Lake Mendota. We collected triplicate samples from each site using a 0.625 m-2 circular quadrat and an airlift method with a modified SCUBA tank suction device called an AquaVac. Air was released through a PVC pipe, creating backpressure to lift sediment, which was captured in a 500μm mesh bag and transported in a resealable plastic bag. We chose an airlift method because of difficulty closing Eckman samplers on the hard substrates of rock and zebra mussel druses. Occasionally additional samples were taken with an Eckman, often at deeper depths, for comparing to the main transects and depths sampled with AquaVac or to collect additional material for isotope analysis.<br/>

Methods

We sampled phytobenthos twice a summer (early June and late August) from 2016-2018 at five depths (1, 3, 5, 8, and 10m) along three transects running perpendicular to shore (A-C, Fig. 1). We collected triplicate samples at each site. SCUBA divers retrieved one rock at rock-dominated sites, or a petri dish full of undisturbed sediment at sand- and muck-dominated sites, and transported samples to the surface in a resealable plastic bag. In the laboratory, we scrubbed phytobenthos from rocks with a brush or emptied petri dish contents into a beaker. We separated phytobenthos from inorganic material by adding ~1 L of deionized water, homogenizing the sample, allowing settlement of inorganic material, and decanting the suspended phytobenthos. We kept samples dark and refrigerated until completely processed to prevent cell division after collection. <br/>We sampled phytobenthos twice a summer (early June and late August) from 2016-2018 at five depths (1, 3, 5, 8, and 10m) along three transects running perpendicular to shore (A-C, Fig. 1). We collected triplicate samples at each site. SCUBA divers retrieved one rock at rock-dominated sites, or a petri dish full of undisturbed sediment at sand- and muck-dominated sites, and transported samples to the surface in a resealable plastic bag. In the laboratory, we scrubbed phytobenthos from rocks with a brush or emptied petri dish contents into a beaker. We separated phytobenthos from inorganic material by adding ~1 L of deionized water, homogenizing the sample, allowing settlement of inorganic material, and decanting the suspended phytobenthos. We kept samples dark and refrigerated until completely processed to prevent cell division after collection.

Methods

I. Study System Lake Mývatn, Iceland (65&deg;36 N, 17&deg;0&prime; W) is a large (38 km²) shallow (4 m max depth) lake divided into two large basins that function mostly as independent hydrologic bodies (Ólafsson 1979). The number of non-biting midge (Diptera: Chironomidae) larvae on the lake bottom is high, but variable: midge production between 1972-74 ranged from 14-100 g ash-free dw m^-2 yr^-1, averaging 28 g dw m^-2 yr^-1 (Lindegaard and Jónasson 1979). The midge assemblage is mostly comprised of two species (&gt; 90% of total individuals), Chironomus islandicus (Kieffer) and Tanytarsus gracilentus (Holmgren) that feed as larvae in the sediment in silken tubes by scraping diatoms, algae, and detritus off the lake bottom (Lindegaard and Jónasson 1979). At maturity (May-August) midge pupae float to the lake surface, emerge as adults, and fly to land, forming large mating swarms around the lake (Einarsson et al. 2004, Gratton et al. 2008). On land, midges are consumed by terrestrial predators (Dreyer et al. 2012, Gratton et al. 2008), or enter the detrital pool upon death (Gratton et al. 2008, Hoekman et al. 2012). Midge populations naturally cycle with 5-8 year periodicity, with abundances fluctuating by 3-4 orders of magnitude (Einarsson et al. 2002, Ives et al. 2008). II. Midge Infall Measurement We deployed eleven transects of passive, lethal aerial infall traps arrayed at variable distances from Lake Mývatn to estimate relative midge abundance on shore during the summers 2008-2011. Each transect was perpendicular to the lake edge, with traps located at approximately 5, 50, 150, and 500 m (where possible) from shore for a total of 31 traps around the lake. Sampling locations were recorded using GPS and precise distances from the lake were calculated within a geographic information system. Traps consisted of a single 1000 mL clear plastic cup (0.0095 m² opening) affixed 1 m above the ground on a stake and filled with 300-500 mL of a 1:1 mixture of water and ethylene glycol and a trace amount of unscented detergent to capture, kill, and preserve insects landing on the surface of the liquid (Gratton et al. 2008, Dreyer et al. 2012). Midges and other insects were emptied from the traps weekly and the traps were reset immediately, thus collections span the entirety of each summer. III. Identification, Counts, and Conversions Midges were counted and identified to morphospecies, small and large. The midge (Diptera,Chrionomidae) assemblage at Mývatn is dominated by two species, Chironomus islandicus (Kieffer)(large, 1.1 mg dw) and Tanytarsus gracilentus (Holmgren)(small, 0.1 mg dw), together comprising 90 percent of total midge abundance (Lindegaard and Jonasson 1979). First, the midges collected in the infall traps were spread out in trays, and counted if there were only a few. Some midges were only identified to the family level of Simuliidae, and other arthropods were counted and categorized as the group, others. Arthropods only identified to the family level Simuliidae or classified as others were not dually counted as Chironomus islandicus or Tanytarsus gracilentus . If there were many midges, generally if there were hundreds to thousands, in an infall trap, subsamples were taken. Subsampling was done using plastic rings that were dropped into the tray. The rings were relatively small compared to the tray, about 2 percent of the area of a tray was represented in a ring. The area inside a ring and the total area of the trays were also measured. Note that different sized rings and trays were used in subsample analysis. These are as follows, trays, small (area of 731 square centimeters), &ldquo;large1&rdquo; (area of 1862.40 square centimeters), and large2 (area of 1247 square centimeters). Rings, standard ring (diameter of 7.30 centimeters, subsample area is 41.85 square centimeters) and small ring (diameter of 6.5 centimeters, subsample area is 33.18 square centimeters). A small ring was only used to subsample trays classified as type &ldquo;large2.&rdquo;The fraction subsampled was then calculated depending on the size of the tray and ring used for the subsample analysis. If the entire tray was counted and no subsampling was done then the fraction subsampled was assigned a value of 1.0. If subsampling was done the fraction subsampled was calculated as the number of subsamples taken multiplied by the fraction of the tray that a subsample ring area covers (number of subsamples multiplied by (ring area divided by tray area)). Note that this is dependent on the tray and ring used for subsample analysis. Finally, the number of midges in an infall trap accounting for subsampling was calculated as the raw count of midges divided by the fraction subsampled (raw count divided by fraction subsampled).Other metrics such as total insects in meters squared per day, and total insect biomass in grams per meter squared day can be calculated with these data. In addition to the estimated average individual midge masses in grams, For 2008 through 2010 average midge masses were calculated as, Tanytarsus equal to .0001104 grams, Chironomus equal to .0010837 grams. For 2011 average midge masses were, Tanytarsus equal to .000182 grams, Chironomus equal to .001268 grams.

Methods

I. Study System Lake Mývatn, Iceland (65&deg;36 N, 17&deg;0&prime; W) is a large (38 km²) shallow (4 m max depth) lake divided into two large basins that function mostly as independent hydrologic bodies (Ólafsson 1979). The number of non-biting midge (Diptera: Chironomidae) larvae on the lake bottom is high, but variable: midge production between 1972-74 ranged from 14-100 g ash-free dw m^-2 yr^-1, averaging 28 g dw m^-2 yr^-1 (Lindegaard and Jónasson 1979). The midge assemblage is mostly comprised of two species (&gt; 90% of total individuals), Chironomus islandicus (Kieffer) and Tanytarsus gracilentus (Holmgren) that feed as larvae in the sediment in silken tubes by scraping diatoms, algae, and detritus off the lake bottom (Lindegaard and Jónasson 1979). At maturity (May-August) midge pupae float to the lake surface, emerge as adults, and fly to land, forming large mating swarms around the lake (Einarsson et al. 2004, Gratton et al. 2008). On land, midges are consumed by terrestrial predators (Dreyer et al. 2012, Gratton et al. 2008), or enter the detrital pool upon death (Gratton et al. 2008, Hoekman et al. 2012). Midge populations naturally cycle with 5-8 year periodicity, with abundances fluctuating by 3-4 orders of magnitude (Einarsson et al. 2002, Ives et al. 2008). II. Midge Emergence Measurement We used submerged conical traps to estimate midge emergence from Lake Mývatn. Traps were constructed of 2 mm clear polycarbonate plastic (Laird Plastics, Madison, WI) formed into a cone with large-diameter opening of 46 cm (0.17 m²). The tops of the cones were open to a diameter of 10 cm, with a clear jar affixed at the apex. The trap was weighted to approximately neutral buoyancy, with the jar at the top containing air to allow mature midges to emerge. Traps were suspended with a nylon line ~1 m below the surface of the lake from an anchored buoy. For sampling, traps were raised to the surface and rapidly inverted, preventing midges from escaping. Jars and traps were thoroughly rinsed with lake water to collect all trapped midges, including unmetamorphosed larvae and pupae, and scrubbed before being returned to the lake to prevent growth of epiphytic algae and colonization by midges. We assume that the emergence traps collect all potentially emerging midges from the sampling area, though it is likely an underestimate, since some midges initially captured could fall out of the trap. Thus, our results should be considered a conservative estimate of potential midge emergence from the surface of the lake.We sampled midge emergence throughout the south basin of Lake Mývatn. Emergence was sampled at six sites in 2008 and 2011 and ten sites in 2009 and 2010, with locations relocated using GPS and natural sightlines. Each site had two traps within 5 m of each other that were monitored during midge activity, approximately from the last week of May to the first week of August. Midge emergence outside of this time frame is extremely low (Lindegaard &amp; Jónasson 1979) and we assume it to be zero. Traps were checked weekly during periods of high emergence (initial and final 2-3 weeks of the study), and bi-weekly during low emergence periods in the middle of the study (July). III. Identification, Counts, and Conversions Midges were counted and identified to morphospecies, small and large. The midge (Diptera,Chrionomidae) assemblage at Mývatn is dominated by two species, Chironomus islandicus (Kieffer)(large, 1.1 mg dw) and Tanytarsus gracilentus (Holmgren)(small, 0.1 mg dw), together comprising 90 percent of total midge abundance (Lindegaard and Jonasson 1979). First, the midges collected in the infall traps were spread out in trays, and counted if there were only a few. Some midges were only identified to the family level of Simuliidae, and other arthropods were counted and categorized as the group, others. Arthropods only identified to the family level Simuliidae or classified as others were not dually counted as Chironomus islandicus or Tanytarsus gracilentus . If there were many midges, generally if there were hundreds to thousands, in an infall trap, subsamples were taken. Subsampling was done using plastic rings that were dropped into the tray. The rings were relatively small compared to the tray, about 2 percent of the area of a tray was represented in a ring. The area inside a ring and the total area of the trays were also measured. Note that different sized rings and trays were used in subsample analysis. These are as follows, trays, small (area of 731 square centimeters), &ldquo;large1&rdquo; (area of 1862.40 square centimeters), and large2 (area of 1247 square centimeters). Rings, standard ring (diameter of 7.30 centimeters, subsample area is 41.85 square centimeters) and small ring (diameter of 6.5 centimeters, subsample area is 33.18 square centimeters). A small ring was only used to subsample trays classified as type &ldquo;large2.&rdquo;The fraction subsampled was then calculated depending on the size of the tray and ring used for the subsample analysis. If the entire tray was counted and no subsampling was done then the fraction subsampled was assigned a value of 1.0. If subsampling was done the fraction subsampled was calculated as the number of subsamples taken multiplied by the fraction of the tray that a subsample ring area covers (number of subsamples multiplied by (ring area divided by tray area)). Note that this is dependent on the tray and ring used for subsample analysis. Finally, the number of midges in an infall trap accounting for subsampling was calculated as the raw count of midges divided by the fraction subsampled (raw count divided by fraction subsampled).Other metrics such as total insects in meters squared per day, and total insect biomass in grams per meter squared day can be calculated with these data. In addition to the estimated average individual midge masses in grams, For 2008 through 2010 average midge masses were calculated as, Tanytarsus equal to .0001104 grams, Chironomus equal to .0010837 grams. For 2011 average midge masses were, Tanytarsus equal to .000182 grams, Chironomus equal to .001268 grams.

Methods

I. Field MethodsThe site where this manipulative field experiment was conducted on the Kalfastrond peninsula at Lake Myvatn is approximately 150 meters long and 75 meters wide. The vegetation consists of grasses Deschampsia spp., Poa spp., and Agrostis spp.), sedges (Carex spp.), and forbs (Ranunculus acris, Geum rivale,and Potentilla palustris). The experimental midge exclusions occurred from the middle or end of May to the beginning of August in each year, the period corresponding to the active growing season of plants and the flight activity of midges at this site. 2 by 2 meter plots were established in 6 blocks spread across the site. Each block included 3 treatment levels, an open control plot, a full exclusion cage and a partial exclusion cage, for a total of 18 experimental plots. Control plots were open to allow complete access to all arthropods. Experimental midge exclusion cages were 1 meter high and constructed from white PVC tubing affixed to rebar posts on each corner of the plot, Plate 1. Full exclusion cages were entirely covered with white polyester netting, 200 holes per square inch, Barre Army Navy Store, Barre VT, USA, to prevent midges from entering the plot. The mesh netting completely enclosed the 2 by 2 by 1 meter frame to prevent flying insects from entering, however the mesh was not secured to the ground in order to allow non flying,ground crawling, arthropods to freely enter and exit the cages. Partial exclusion cages had one 0.5 meter strip of mesh stretched around the outside of the frame and another 0.75 meter strip draped over the top. Partial cages were designed to examine any effects of cages themselves on terrestrial responses while minimally affecting midge inputs into the plots and arthropod movement.The partial exclusion treatment was discontinued in 2011. Each plot contains a pitfall and an infall trap that are continuously sampled during the summer, while the cages are up. Vacuum samples were taken from the plots about once per month in 2008 through 2010 and only once per summer for subsequent summers.Midge activity was measured in all plots to document changes in midge abundance over the course of a season and between years and to assess the degree to which cages excluded midges. Midge abundance in the plots was continuously measured using passive aerial infall traps consisting of a 1000 milliliter clear plastic cup, 95 square centimeter opening, attached to a post 0.5 meters high and filled with 250 milliliters of a 1 to 1 ethylene glycol to water solution and a small amount of unscented detergent to capture and kill insects that alighted upon the surface. Infall traps were emptied about every 10 days.II. AnalysisMidges were counted and identified to morphospecies, small and large. The midge (Diptera,Chrionomidae) assemblage at Myvatn is dominated by two species,Chironomus islandicus (Kieffer)(large, 1.1 mg dw) and Tanytarsus gracilentus(Holmgren)(small, 0.1 mg dw), together comprising 90 percent of total midge abundance (Lindegaard and Jonasson 1979). First, the midges collected in the infall traps were spread out in trays, and counted if there were only a few. Some midges were only identified to the family level of Simuliidae,and other arthropods were counted and categorized as the group, others. Arthropods only identified to the family level Simuliidae or classified as others were not dually counted as Chironomus islandicus or Tanytarsus gracilentus. If there were many midges, generally if there were hundreds to thousands, in an infall trap,subsamples were taken. Subsampling was done using plastic rings that were dropped into the tray. The rings were relatively small compared to the tray, about 2 percent of the area of a tray was represented in a ring. The area inside a ring and the total area of the trays were also measured. Note that different sized rings and trays were used in subsample analysis. These are as follows, Trays, small (area of 731 square centimeters), large1 (area of 1862.40 square centimeters), and large2 (area of 1247 square centimeters). Rings, standard ring (diameter of 7.30 centimeters, subsample area is 41.85 square centimeters) and small ring (diameter of 6.5 centimeters, subsample area is 33.18 square centimeters). A small ring was only used to subsample trays classified as type large2.The fraction subsampled was then calculated depending on the size of the tray and ring used for the subsample analysis. If the entire tray was counted and no subsampling was done then the fraction subsampled was assigned a value of 1.0. If subsampling was done the fraction subsampled was calculated as the number of subsamples taken multiplied by the fraction of the tray that a subsample ring area covers (number of subsamples multiplied by (ring area divided by tray area)). Note that this is dependent on the tray and ring used for subsample analysis. Finally, the number of midges in an infall trap accounting for subsampling was calculated as the raw count of midges divided by the fraction subsampled (raw count divided by fraction subsampled).Other metrics such as total insects in meters squared per day, and total insect biomass in grams per meter squared day can be calculated with these data. in addition to the estimated average individual midge masses in grams, For 2008 through 2010 average midge masses were calculated as, Tanytarsus equal to .0001104 grams, Chironomus equal to .0010837 grams. For 2011 average midge masses were, Tanytarsus equal to .000182 grams, Chironomus equal to .001268 grams.