Cayla L. Carlson Jeffrey C. Jolley Peter Sakaris 20260422 tabular digital data Reston, VA U.S. Geological Survey Authors Open Researcher and Contributor Id (ORCID) are as follows: Cayla L. Carlson: 0000-0002-4609-8731; Jeffrey C. Jolley: 0000-0002-4674-5126; Peter Sakaris:0000-0003-1122-4272 https://doi.org/10.5066/P136VW5B Accurate age estimation is crucial for understanding fish population dynamics, yet daily age validation for invasive Grass Carp in the Midwestern U.S. is lacking. We validated daily otolith increment deposition in known-age Grass Carp reared in laboratory and pond environments. Larval Grass Carp from captive broodstock were sampled between 6- and 120-days post-hatch. Lapilli otoliths were extracted, lightly sanded, and read three times to estimate age, in days, which was compared against known age. Daily increment deposition was validated through 51 days post-hatch for tank-reared fish and 69 days for pond-reared fish, indicating lapilli otoliths can provide reliable daily age estimates for early life stages of Grass Carp. These validated aging methods support studies of early growth, recruitment, and dispersal and provide critical information for understanding and managing invasive Grass Carp populations. 20180726 20181108 ground condition Complete None planned Columbia Environmental Research Center -92.284312 -92.274450 38.912612 38.910784 ISO 19115 Topic Category biota USGS Thesaurus fish invasive species National Agricultural Library Thesaurus age determination otoliths USGS Metadata Identifier USGS:69e7f3dbb66b0164d0f730c3 Geographic Names Information System Missouri None Columbia Environmental Research Center Integrated Taxonomic Information System Ctenopharyngodon idella Kingdom Animalia animals Subkingdom Bilateria triploblasts Infrakingdom Deuterostomia Phylum Chordata chordates Subphylum Vertebrata vertebrates Infraphylum Gnathostomata Superclass Actinopterygii ray-finned fishes spiny rayed fishes Class Teleostei Superorder Ostariophysi Order Cypriniformes minnows suckers Superfamily Cyprinoidea Family Cyprinidae carps minnows carps and minnows Genus Ctenopharyngodon grass carps Species Ctenopharyngodon idella grass carp silver orfe TSN: 163537 None. Please see 'Distribution Info' for details. These data are marked with a Creative Common CC0 1.0 Universal License. These data are in the public domain and do not have any use constraints. It is requested that the authors be cited for any subsequent publications that reference this dataset. Users are advised to read the dataset's metadata thoroughly to understand appropriate use and data limitations. Cayla L Carlson USGS - SOUTHEAST REGION Fish Biologist mailing and physical

Columbia Environmental Res Ctr,CERC - R1 Research Building

Columbia MO 65201 573-875-5399 clcarlson@usgs.gov Windows 11 version 23H2, build 22631.6649; notepad++ 64-bit x64 version 8.9.2 Scales used to measure fish weight are calibrated yearly to ensure accuracy. Microscopes used to measure otolith diameters were calibrated using a calibration slide before each use. Data was entered and verified. All values were within expected ranges. Otoliths that were considered unreadable by otolith processor Pete Sakaris were excluded. Otherwise, data set is considered complete for the information presented, as described in the abstract. Users are advised to read the rest of the metadata record carefully for additional details. No formal positional accuracy tests were conducted. No formal positional accuracy tests were conducted. Captive Grass Carp broodstock were hormonally induced and spawned in July of 2018 at the United States Geological Survey Columbia Environmental Research Center in north-central Missouri. These known-age Grass Carp larvae were hatched on July 10th, 2018, and initially reared in multiple 15-gallon aquaria maintained at 21–22°C under a 12-hour light, 12-hour dark photoperiod. Tank-reared fish were fed brine shrimp three times daily, with flake food gradually introduced. At 9 days post-hatch (July 19th, 2018), a subset of larvae was transferred to three outdoor rearing ponds. Pond-reared fish were fed pelleted Rangen fish food once daily until late October. The pond was also fertilized monthly with inorganic fertilizer and soybean meal to enhance natural productivity. 20180710 Grass Carp were sampled from aquaria between 6- and 120-days post-hatch and from the pond between 16- and 120-days post-hatch at intervals of 5–15 days. Sampling intervals were intentionally varied to minimize potential aging bias associated with regular sampling schedules. After samples were collected, fish were euthanized using a lethal dose of MS-222, they were then measured for total length (mm) and weight (g). Whole fish samples were then sent to our project partner at Georgia Gwinnett College in Lawrenceville, Georgia. This is where the lapilli otoliths were extracted under a dissecting microscope and mounted directly on microscope slides and fixed with clear epoxy resin (105 Epoxy Resin, 206 Slow Hardener, West System, Bay City, Michigan). Otoliths were then lightly sanded using 1200-grit sandpaper to expose the core and daily increments. 2018 Prepared sections were examined under a compound microscope at 100x to 400x magnification using image analysis software (i-solution Lite software, IMT i-Solution Inc. Riverton, Utah). Daily increments were counted from the core to the outer margin along a consistent transect. Each otolith was read three times in random order, with no two consecutive reads from the same otolith. Otolith radius and long-axis diameter were measured for each section. 2019 Grass_Carp_Daily_Aging.csv Comma Separated Value (CSV) file containing Estimates of daily age from lapilli otolith ring counts. Data include known age, length, and weight of sampled fish; age estimates from otolith ring counts (3 per otolith) and otolith size data. Producer Defined system The system in which the fish was reared from 9 days post hatch until sampling. Producer Defined Pond Fish was sampled from outdoor rearing ponds. Pond-reared fish were fed pelleted Rangen fish food once daily until late October. Producer defined Tank Fish was sampled from 15-gallon aquaria maintained indoors at 21–22°C under a 12-hour light, 12-hour dark photoperiod. Tank-reared fish were fed brine shrimp three times daily, with flake food gradually introduced. Producer defined fish_id A unique alphanumeric identifier assigned to each fish and the otolith from that fish. Producer Defined PGC (pond grass carp) or TGC (Tank grass carp) followed by a hyphen and three-digit number. harvest_date The date the fish was harvested, expressed as YYYYMMDD. Producer Defined 20180726 20181108 YYYYMMDD DPH days post hatch. The known age of the fish, in days post hatch, at the time the fish was harvested. Producer Defined 16 120 days TL The total length of the fish, in millimeters, measured from the tip of the snout to the tip of the tail. Producer Defined 9 73 millimeters weight The wet weight of the fish at time of harvest, in grams. Producer Defined NR No record, typically due to fish being too small to register on gram scale. Producer defined 0.01 4.4 grams read_1 The first age estimate. The estimated age of the fish, in days post hatch, based on the otolith ring count. Each otolith was read in triplicate. Producer Defined NR No record. This could be due to otolith getting lost during processing OR otolith unable to be read due to legibility of daily rings so batch was excluded. Producer defined 19 120 days post hatch read_2 The second age estimate. The estimated age of the fish, in days post hatch, based on the otolith ring count. Each otolith was read in triplicate. Producer Defined NR No record. This could be due to otolith getting lost during processing OR otolith unable to be read due to legibility of daily rings so batch was excluded. Producer defined 19 124 days post hatch read_3 The third age estimate. The estimated age of the fish, in days post hatch, based on the otolith ring count. Each otolith was read in triplicate. Producer Defined NR No record. This could be due to otolith getting lost during processing OR otolith unable to be read due to legibility of daily rings so batch was excluded. Producer defined 19 123 days post hatch diameter The diameter of the longest axis of the otolith, in millimeters Producer Defined NR No record Producer defined 0.14 1.13 millimeters long_radius The radius of the otolith, measured from core to long edge, in millimeters Producer Defined NR No record Producer defined 0.08 0.78 millimeters This data release is comprised of one comma-separated value (CSV) file which contains age estimates for each sampled fish. Carlson, C.L., Jolley, J.C., and Sakaris, P., 2026, Validating daily age via lapilli otolith growth rings in larval Grass Carp Ctenopharyngodon idella: U.S. Geological Survey data release, https://doi.org/10.5066/P136VW5B. GS ScienceBase U.S. Geological Survey mailing and physical

Denver Federal Center, Building 810, Mail Stop 302

Denver CO 80225 United States 1-888-275-8747 sciencebase@usgs.gov Unless otherwise stated, all data, metadata and related materials are considered to satisfy the quality standards relative to the purpose for which the data were collected. Although these data and associated metadata have been reviewed for accuracy and completeness and approved for release by the U.S. Geological Survey (USGS), no warranty expressed or implied is made regarding the display or utility of the data on any other system or for general or scientific purposes, nor shall the act of distribution constitute any such warranty. Any use of trade, firm, or product names is for descriptive purposes only and does not imply endorsement by the U.S. Government. Digital Data https://doi.org/10.5066/P136VW5B None 20260422 CERC Data Manager U.S. Geological Survey, Columbia Environmental Research Center Natural Resource Data Manager mailing and physical

4200 New Haven Road

Columbia MO 65201 United States 573-875-5399 gs-mw-cerc_data_manager@usgs.gov FGDC Biological Data Profile of the Content Standard for Digital Geospatial Metadata FGDC-STD-001.1-1999