DOI: 10.6084/m9.figshare.13683910.v1
title: Larval anuran developmental stoichiometry and areal biomass from geographically isolated wetlands in Georgia, USA

description: Raw data on larval anuran developmental stoichiometry across 11 species (Knapp_et_al_Stoichiometry_Dataset.xlsx) and larval anuran biomass data from four wetlands collected through throw sampling (Knapp_et_al_Biomass_Dataset.xlsx). All animal handling in this study was performed under The University of Alabama Institutional Animal Care and Use Committee protocol ID 15-10-0110.<br><i>Study site</i> We conducted this study at the Jones Center at Ichauway (hereafter Ichauway) located in Baker County, Georgia, USA. <br><i>Larval anuran collection</i> We used a dip net (3-mm mesh size) to collect larval anurans intermittently across the hydroperiod from February 2017 to September 2018 at the four study GIWs. We timed our sampling to collect samples of all available larvae across five developmental stages. The five stages were simplified [Column = Simplified Stages] as follows based on Gosner (1960): Stage 1) no limbs (Gosner stages 23-25), Stage 1.5) developing hindlimbs (Gosner stages 26-37), Stage 2) fully-developed hindlimbs (Gosner stages 38-40), Stage 2.5) developing forelimbs (Gosner stage 41), and Stage 3) developed forelimbs, but retaining a tail (Gosner stages 42-46; Fig. 1). We euthanized larval anurans using 5 g/L tricaine methanesulfonate (MS-222) solution and froze them at -20 °C for later stoichiometric analysis. <i><br></i><i>Stoichiometric analysis</i> We dissected euthanized larvae to remove the gastrointestinal tract below the manicotto glandulare to the rectum, measured total length (body plus tail) to the nearest 1 mm, and weighed larvae to the nearest 0.001 g. Whole-body (body and tail) larval anurans were freeze-dried for ≥ 72 h, and then re-weighed to determine larval dry-mass. We used either a mortar and pestle or a Retsch Mixer Mill to homogenize dried larvae. To measure total %C and total %N tissue content, we analyzed a ~1-3 mg subset of each homogenized larva with a FLASH 2000 NC Analyzer. We combusted a further ~1-3 mg subset of each homogenized larva in a Thermolyne muffle furnace at 500 °C for 1 h, digested each in 1 N HCl at 105 °C for 2 h, and used the ascorbic acid-molybdate method to determine total %P. Larvae &lt; 4 mg dry-mass were homogenized together with like-species and stages collected from the same wetland to increase particulate material for our analysis.<br><i>Larval anuran density, biomass, and nutrient storage sampling</i><i></i> We measured biomass of larval anurans monthly during the hydroperiods (March to June 2018) of the four study wetlands using an aluminum 0.37 m<sup>2</sup> throw trap. We sampled a minimum of 0.025% of the wetland surface area (calculated from wetland contour data and water-level data [Jones Center at Ichauway, unpublished data]); throws were concentrated in water &lt; 0.5 m in depth. For each throw, larval anurans were removed from the trap using a dip net (3-mm mesh size) and we continued dip netting until we had ten consecutive sweeps without captures. Each larva was identified to species, measured for total length to the nearest 1-mm, weighed to the nearest 0.01-g, assigned to one of five developmental stage categories as described above [Column = Simplified Stages], and released.