README Welcome! Thank you for your interest in our data. You can contact the corresponding author for questions: Nadine-Cyra Freistetter nadine.freistetter@fmi.fi (last updated 2022) The file Freistetter_et_al_2022_IsotopicAnalysis_Figures_LymantriaDispar.xlsx contains all measurement and analysis data of the natal provenance analysis, including figures, The file Freistetter_et_al_2022_Equilibriation_Experiments1_LymantriaDispar_Pex.xlsx contains the results to the equilibration experiments carried out to determine the Percentage exchangable hydrogen (Pex). The results of both files are presented in the publication Freistetter et al. (2022) "Tracking Global Invasion Pathways of the Spongy Moth (Lepidoptera: Erebidae) to the U.S. using Stable Isotopes as Endogenous Biomarkers" published in Ecology and Evolution. The contents of both files are made available under the Open Data Commons Attribution License: http://opendatacommons.org/licenses/by/1.0/ Please always correctly cite the paper when referring to or using the data provided here. The analyses were done with the Microsoft 365 Excel 64-bit version. All sheets and the whole work book are protected. To unprotect them, go to the menu tab "Review" and in the "Protect" section, click "Unprotect Sheet" and "Unportect Workbook". You require no password. We will now go through each sheet of the files. * * * - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - * * * * * * - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - * * * FILE: Freistetter_et_al_2022_IsotopicAnalysis_Figures_LymantriaDispar.xlsx EXCEL SHEETS CONTAINED IN THE FILE: - "Samples Left" - "Pex Sensitivity" - "Calibration" - "Egg-to-Adult" - "Hungate" - "ClimChngCorr." - "World Model" - "Data-Controls" - "Data-Moths" - "Data-Eggs" - "Fig0" - "Fig1" - "Fig2" - "Fig3" - "Fig4" - "Fig5a" - "Fig5b" Following, we will go through the excel sheets one by one. *************************** SHEET "SAMPLES LEFT" The samples were recieved from die USDA in 2017 in 4 compartmentarized cardboard boxes, referred to as BOX #1, BOX #2, BOX #3 and BOX #4 in the sheet. Each box-compartment had one platic vile with a sample and one or two silica gel beads. Columns: - SPECIMEN: The samples were named by the USDA lab. The first two or three letters refer to the country the sample was collected in. All geographic information can be found in the "Data" Sheets (Data-Moths, Data-Eggs, Data-Controls) - SAMPLE: After all measurements, we noted, which samples contained enough material to be measured in another Deuterium measurement campaign (with sample size of 0.2mg). "Enough" sample left means more than 0.2mg of sample left. "Little" means less than 0.2mg of sample left. "None" means no sample left. Assessment done in 2018. The stock might have changed by now. Contact rebecca.hood-nowotny@boku.ac.at for current information. - COMMENTS: If the sample contained anything else than moth feet, it was marked in the *************************** SHEET "Pex Sensitivity" We assumed different possible values for percentage exchangeable hydrogen (Pex or P_ex) which were obtained from literature and tested how much it would change the natal signature. The figure in the sheet is not shown in the publication. *************************** SHEET "Calibration" Measurements are always subject to random measurement errors. It is a standard procedure in chemical analysis to correct the measurement values for these errors by means of a calibration curve. For that, reference materials real (expected) values are compared to their measured values on a x-y-plane. The trend curve through the points is the calibration curve that is being used to correct the sample values for measurement errors. There were a total of 6 measurements done (5 of which from the main author, therefore the sixth is marked differently). Lines 4-10 show the measurement dates, the intercept and slope of the calibration curve as well as measurement accuracy and which isotopes were measured: hydrogen (H) or nitrogen (N). *************************** SHEET "Egg-to-Adult" The egg-to-adult conversion factor was calculated by simply calculating the difference in isotopic value between the control moths reared at the insectary at the USDA. This value (marked in red) was later-on used on eggs to use the same provenance analysis like for adult moth samples. *************************** SHEET "Hungate" Hungate et al. (2016) (https://doi.org/10.1371/journal.pone.0149599) had shown a local linear correlation between the hydrogen isotopic values of japanese beetles and those of the precipitation on-site. Since the correlation curve formula was not given in the publication, we used the figure provided in the paper (left figure in the sheet) to roughly reconstruct the formula and use our values regressed with this formula as "(Hungate) expected insect values". The lines 3 and 4 show point 1 (P1) and point 2 (P2) reconstructed coordinates and in a thick black frame next to it, the resulting intercept (k) and slope (d) of a line between those two points (right figure in the sheet). *************************** SHEET "ClimChngCorr." = Climate Change Correction The values for expected presecipitation hydrogen isotope values used in our study were taken from the OIPC (Online Isotopes in Precipitation Calculator). These values are strongly dependent on climate, influenced by temperatures and precipitation. The OIPC is calibrated to today's climate. However, some samples were from the 1990s. So we compared average temperatures around 1990 with those of today for a handful of place from which we had samples. With the help of Liu et al. (2018) we calculated the shift in hydrogen isotopes due to a changed temperature. The "Climate Coefficient" (in red) obtained was used as an offset for the natal values of samples that were collected before 2000. *************************** SHEET "World Model" Here, the slope (k) and intercept (d) of the world regression models obtained from Figure 3 are summarized and highlighted by a red frame. Underneath, two figures showing the isoscapes of hydrogen (Terzer et al. 2013) and nitrogen (Bowen & West 2008) isotopes are used to create the isotopic zones for our study. *************************** SHEETS "Data-Controls", "Data-Moths" and "Data-Eggs" We finally come to our actual data. All "Data-..." sheets are structured in the same way. Columns A-P : First, we have the sample number and various data and characteristics of the place the sample is from. Columns Q-W : Following are details to the sample, the USDA identifier, collection date of the sample, storage box etc. Column X : If the genetic strain of the moth was known (e.g. through measurement by USDA, methods not shown), it was marked here. Column Z : Hydrogen isotopic composition of water in air in the laboratory on the day of measurment for the purpose of corrections Columns AA-BB: Full analysis of the HYDROGEN isotopic measurements. The column headers describe each step. Columns AR-AT try to detect exotic moths Columns AU-BA try to locate the moths' natal origins. Columns BD-CA: Full analysis of the NITROGEN isotopic measurements. The column headers describe each step. Columns BR-BX try to detect exotic moths. Columns BY-CA try to locate the moths' natal origins. In the "Data-Controls" sheet, we present and analyse the control specimens, eggs and moths reared under controlled conditions in the USDA insectary. In the "Data-Moths" sheet, we present and analyse the field-captured adult specimens. In the "Data-Eggs" sheet, we present and analyse the field-captured eggs. *************************** SHEETS "Fig0" - "Fig6b" The data used for the figures is displayed in the according sheets and automaticall calculated in excel via the LOOKUP function from the "Data-..." sheets. * * * - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - * * * * * * - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - * * * FILE: Freistetter_et_al_2022_Equilibriation_Experiments1_LymantriaDispar_Pex.xlsx EXCEL SHEETS CONTAINED IN THE FILE: - "raw_data" - "Calibration" - "Pex" - "Natal" - "Summary" Following, we will go through the excel sheets one by one. *************************** SHEET "raw_data" Data as produced by the measurement instrument. The columns "nominated" (J) and "%exch." (K) were added by us. The nominated value is the value that the reference materials are supposed to have (which might not be the measured values due to random measurement errors). The percentage exchangeable hydrogen ("%exch.") is the share of hydrogen atoms the reference materials (blue background) reportedly exchanges with hydrogen atoms from the ambient air. There were two groups of samples ("GR1" and "GR2") that were equilibrated with two distinct atmospheres (distinct in their isotopic composition). By help of the measurement results and a simple formula, we can calculate the percentage exchangeable hydrogen of the moth samples. The moths were samples from Wisconsin (therefore the "WI" in the identifier). The numbers are unimportant. USGS is a reference material. For comparison with existing literature on comparative equilibration experiments, also japanese beetle (JapBeetle) samples were equilibrated. *************************** SHEET "Calibration" Measurements are always subject to random measurement errors. It is a standard procedure in chemical analysis to correct the measurement values for these errors by means of a calibration curve. For that, reference materials real (expected) values are compared to their measured values on a x-y-plane. The trend curve through the points is the calibration curve that is being used to correct the sample values for measurement errors. *************************** SHEET "Pex" Cells A2 and K2 show the hydrogen isotopic values of the vapourized water the samples were equilibrated with for two weeks. Columns B-H show the measurement results for Group 1. Columns L-R show the measurement results for Group 2. Column J shows the difference in isotopic value between the corresponding samples (mostly from the very same leg or at least from the same colony). Lines 35-55 shows the results of the percentage exchangable hydrogen obtained from the experiment. *************************** SHEET "Natal" The natal values (eliminated Pex) were calculated and compared for the two groups. There were indications that some samples were slightly wet. The equilibration experiment was therefore repeated two more times, but we always obtained the same Pex. *************************** SHEET "Summary" The findings of the equilibration experiments are summed up in a table. * * * - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - * * * * * * - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - * * * The results of both files are presented in the publication Freistetter et al. (2022) "Tracking Global Invasion Pathways of the Spongy Moth (Lepidoptera: Erebidae) to the U.S. using Stable Isotopes as Endogenous Biomarkers" published in Ecology and Evolution. The contents of both files are made available under the Open Data Commons Attribution License: http://opendatacommons.org/licenses/by/1.0/ Please always correctly cite the paper when referring to or using the data provided here. Thank you for your interest in our data. You can contact the corresponding author for questions: Nadine-Cyra Freistetter nadine.freistetter@fmi.fi (last updated 2022)