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Experimental setup:<\/strong> Two complementary experiments were conducted in late 2018 (E1) and 2020 (E2), each rearing newly fertilized sand lance embryos to hatch over the course of 32-65 days. Founder adults were sampled at Stellwagen Bank National Marine Sanctuary (SBNMS) (42\u00b0 9' 58.26\" N, 70\u00b0 18' 44.19\" W) at the peak of their narrow, local spawning window on November 15th (E1) or 27th (E2), using a 1.3 \u00d7 0.7 m beam trawl (6 mm mesh) towed over ground at 3 knots for 15 min. On deck, all flowing-ripe males and females were strip-spawned together (at 10\u00b0C, E1: Nmale\/female = 29\/13; E2: Nmale\/female = 50\/46) and their progeny transported to the University of Connecticut's Rankin Seawater Lab. There, exposure experiments commenced within 8 hours post fertilization by placing a volumetrically measured random sample of 600 (E1) or 1,200 embryos (E2) into each replicate rearing container.<\/p>\n Experimental seawater was drawn from subsurface eastern Long Island Sound (~ 31 psu), filtered to 1 \u00b5m, and UV-sterilized before use. Oxygen levels were maintained at ~100% saturation, while the photoperiod was 11L:13D.<\/p>\n Seawater chemistry<\/strong>: Realized pCO\u2082 conditions and other seawater chemistry parameters (Table 1 of Baumann et al., MEPS (in review)) were estimated in CO2SYS (V2.1, Pierrot et al. 2006) based on samples taken every 10 days and measured for temperature, pHNIST, salinity (refractometer) and total alkalinity (AT, \u03bcmol kg-1). Seawater samples were filtered to 10 \u00b5m, stored in 300 ml borosilicate bottles at 3\u00b0C, and within days measured for AT using endpoint titration (Mettler Toledo\u00ae G20 Potentiometric Titrator) with an accuracy of \u00b11% (Murray et al. 2019; verified and calibrated using Dr. Andrew Dickson\u2019s certified reference material for AT in seawater; Scripps Institution of Oceanography, Batch Nr. 162 & 164).<\/p>\n Experimental designs:<\/strong> During E1, we tested factorial combinations of two static temperatures and three target pCO\u2082 levels, thereby encompassing contemporary thermal conditions on Stellwagen Bank between late fall (10\u00b0C) and early winter (6\u00b0C), as well as current ambient (400 \u00b5atm, pH~8.12), predicted end-of-century (1,000 \u00b5atm, pH~7.76), and maximum open ocean pCO\u2082 benchmarks (2,000 \u00b5atm, pH~7.48; Caldeira & Wickett 2003, Salisbury & J\u00f6nsson 2018). At 10\u00b0C, three additional, pCO\u2082 levels below 1,000 \u00b5atm (570, 690, 890 \u00b5atm, Table 1) were included to better describe near future CO\u2082 sensitivities of sand lance embryos. The replication level for each of the 9 treatments was N = 5. Another 50 embryos per replicate were subsampled 90-190 ddpf and preserved in buffered (sodium tetraborate) 5% formaldehyde-in-freshwater solution. Those embryos sampled just before hatching began (170 ddpf, one random replicate per treatment) were later submitted for sectioning and staining (H&E stain; Horus Scientific, Worcester, MA), and later imaged for analyses of chorionic thickness (Nikon SMZ-1000 with Luminera\u00ae Infinity2-2 camera and ImagePro Premier v9.0, Media Cybernetics\u00ae).<\/p>\n During E2, we again tested target pCO\u2082 levels of 400, 1,000, and 2,000 \u00b5atm, first at an intermediate static temperature of 7\u00b0C and second at a dynamic temperature of 10\u00b0C decreasing to 5\u00b0C at a rate of 0.2\u00b0C d\u207b\u00b9 (105\u00b0C). The latter was chosen to approximate the seasonal decline in bottom temperatures experienced by sand lance embryos on Stellwagen Bank. The two treatments reached thermal equivalence at 32 dpf (224 ddfp) \u2013 just after hatching had started. To better describe sand lance upper CO\u2082-sensitivities (1,000 - 2,000 \u00b5atm), we added two intermediate pCO\u2082 levels at 7\u00b0C (~1,300; ~1,700 \u00b5atm) and one at 105\u00b0C (~1,300 \u00b5atm). The initial replication level for each of the 9 treatments was N = 6. However, to disentangle potential pCO\u2082 effects on embryonic development vs. effects on hatching itself, we switched three random replicates from each extreme pCO\u2082 treatment per temperature with the opposite pCO\u2082 treatment (i.e., 3\u00d7 ~400 -->\u00a0~2,000 \u00b5atm and 3\u00d7 ~2,000\u00a0-->\u00a0~400 \u00b5atm). The switch happened at 175 ddpf (25 dpf at 7\u00b0C; 22dpf at 105\u00b0C) just before hatching started.<\/p>\n Response traits:<\/strong> From 90 ddpf onwards, rearing containers were monitored daily until hatching commenced; then, the number of hatchlings per replicate was recorded daily until hatching ceased. All hatchlings were immediately preserved in buffered 5% formaldehyde\/freshwater solution for later morphological measurements. At the conclusion of E1, unhatched remains were imaged at 4\u00d7 magnification, allowing the later distinction between (a) early arrested embryos (no or only amorphous cell masses visible), (b) partially developed embryos (unpigmented eyes visible, body not fully wrapped around the egg), and (c) fully developed embryos (pigmented eyes, body clearly visible and more than 1\u00d7 wrapped around; Fig.S2). In E2, we continued daily monitoring for 7 more days after hatching had ceased; then examined the remains microscopically for embryos still alive (i.e., with beating hearts). Absolute hatching numbers were transformed to daily relative frequencies via dividing by the initial number of embryos that was adjusted for subsampling (E1, N = 500 per replicate) or reduced fertilization success (E2, N = 873 per replicate, based on examining independent post-fertilization subsamples). Relative frequencies were then summed to yield cumulative hatching success (HS, %) for each replicate. For E1, we additionally calculated the proportions of (a) fully developed but unhatched embryos and (b) all other arrested embryos combined. The latter also included decayed stages that were no longer detectable at the conclusion of E1.\u00a0<\/p>\n To characterize hatching phenology, we recorded the day of first hatch (dpf), day of peak hatch (= dpf with the highest relative hatch frequency), and the total hatching period (d) for each replicate. Following Murray et al. (2019), a large number of hatchlings were imaged at 4\u00d7 magnification (E1: Ntotal = 3,923; E2: Ntotal = 2,659) and then individually measured (ImagePro) for three morphological traits, i.e., standard length (SL, nearest 0.01 mm), yolk sac area (nearest 0.001 mm\u00b2), and the size of the remaining oil globule inside the yolk sac (nearest 0.001 mm\u00b2). The latter two traits are proxies for endogenous energy reserves after hatching, but they were strongly correlated (N = 5,552; R = 0.62, p < 0.001). Hence, we used PCA to extract the first principal component (explaining 73% (E1), 81% (E2) of variability) and then used the PC1 scores as the new variable, hereafter referred to as \u2018Endogenous Energy Reserves\u2019 (EER). Histological sections of fully developed, pre-hatch embryos from E1 were imaged at 20\u00d7 magnification to measure the thickness of the chorion (ImagePro). Chorion thickness was measured at 10 randomly selected locations around the circumference of the sectioned embryo, with measurements averaged subsequently for each embryo. Unfortunately, fewer than expected embryos were sectioned well enough for quality measurements, ranging from 2-7 per treatment.<\/p><\/div>","@type":"rdf:HTML"}],"http:\/\/ocean-data.org\/schema\/hasBriefDescription":[{"@value":"Hatch Source E1E2","@language":"en-US"}],"http:\/\/www.w3.org\/2000\/01\/rdf-schema#label":[{"@value":"Hatch Source E1E2","@type":"xsd:string"}],"http:\/\/ocean-data.org\/schema\/hasProcessingDescription":[{"@value":" BCO-DMO Processing:<\/strong>
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Founder adults were sampled at Stellwagen Bank National Marine Sanctuary (SBNMS) (42\u00b0 9' 58.26\" N, 70\u00b0 18' 44.1\" W). Two complementary experiments were conducted in late 2018 (E1) and 2020 (E2), each rearing newly fertilized sand lance embryos to hatch over the course of 32-65 days. This dataset includes information on rearing conditions, hatch frequency, and hatch success.","@language":"en-US"}],"http:\/\/purl.org\/dc\/terms\/rights":[{"@id":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"http:\/\/ocean-data.org\/schema\/deprecated":[{"@value":"false","@type":"xsd:boolean"}],"http:\/\/ocean-data.org\/schema\/temporalExtent":[{"@value":"_:temporalExtent867401"}],"http:\/\/ocean-data.org\/schema\/spatialCoverage":[{"@value":"_:spatialCoverage867401"}],"http:\/\/purl.org\/dc\/terms\/bibliographicCitation":[{"@value":"Baumann, H., Nye, J. (2022) Hatch data from experiments on CO2 sensitivity of Northern sand lance (Ammodytes dubius) embryos conducted in 2018 and 2020. 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