# The effect of urbanization and temperature on thermal tolerance, foraging performance, and competition in cavity dwelling ants Human disturbance including rapid urbanization and increased temperatures can have profound effects on the ecology of local populations. Eusocial insects, such as ants, have adapted to stressors of increasing temperature and urbanization, however these evolutionary responses are not consistent among populations across geographic space. Here we asked how urbanization and incubation temperature influence critical thermal maximum (CTmax) and various ecologically relevant behaviors in three ant species in urban and rural locations in Worcester, MA, USA. We did this by incubating colonies of three species of cavity dwelling ant (*Aphaenogaster picea, Tapinoma sessile,* and *Temnothorax longispinosus*) from 2 habitat types (Rural and Urban), for 60-days at multiple temperatures. Included here are the .csv files with data from the project and r code for the analysis. DATA & FILE OVERVIEW 1. File List: A) AcornAntE_E_2023.R B) Ants_Final_CoEdgeACT_RR.csv C) Colony_Final_CoEdge_RR.csv D) EE_Mantel_Test_AP_2.csv E) EE_Mantel_Test_TL_2.csv F) EE_Mantel_TS_2.csv \######################################################################### DATA-SPECIFIC INFORMATION FOR: AcornAntE_E_2023.R This is the R code used for the statistical analysis of the data within files below ##### DATA-SPECIFIC INFORMATION FOR: Ants_Final_CoEdgeACT_RR.csv Data File Description: Individual Ant performance assay data Variable List: * Ant: Individual Ant ID * Col: Colony Number * Species: Ant Species * Collection Location: Site where ant colony was collected * us: Urbanization Status * Incubation date: Date when incubation of colony began * Incubation Temperature: Temperature in Celsius of Incubator * aggression: Defense Index Score * CTmax: Critical thermal maximum of ant ##### DATA-SPECIFIC INFORMATION FOR: Colony_Final_CoEdge_RR.csv Data File Description: Colony level performance assay data Variable List: * colony: Colony Number * Species: Ant Species * Nativity Status: Whether ant species is native * Colony Type: Queen present or absent from colony fragment at collection * Collection Location: Site where ant colony was collected * Urbanization: Urbanization Status * Incubation date: Date when incubation of colony began * Incubation: Temperature in Celsius of Incubator * crmc: Colonization metric corrected for colony size * rr: Recruitment Rate * rr1: Recruitment Rate+1 * AI: Average defense index score per colony * CTmax: Critical thermal maximum of ant ##### DATA-SPECIFIC INFORMATION FOR: EE_Mantel_Test_AP_2.csv Data File Description: Mantel test data for Aphaenogaster picea ants Variable List: * Site: Collection Location * DF: Average Defense Index Score * cc: Average uncorrected colonization rate score * rr: Average recruitment rate score * Lat: Latitude of Site * Long: Longitude of Site * Ctmax: Average critical thermal maximum score * crcm: Average colonization rate score corrected by colony size ##### DATA-SPECIFIC INFORMATION FOR: EE_Mantel_Test_TL_2.csv Data File Description: Mantel test data for Temnothorax longispinosus ants Variable List: * Site: Collection Location * AG: Average Defense Index Score * cc: Average uncorrected colonization rate score * rr: Average recruitment rate score * Lat: Latitude of Site * Long: Longitude of Site * Ctmax: Average critical thermal maximum score * crcm: Average colonization rate score corrected by colony size ##### DATA-SPECIFIC INFORMATION FOR: EE_Mantel_TS_2.csv Data File Description: Mantel test data for Tapinoma sessile ants Variable List: * Site: Collection Location * AG: Average Defense Index Score * Ctmax: Average critical thermal maximum score * rr: Average recruitment rate score * Lat: Latitude of Site * Long: Longitude of Site * crcm: Average colonization rate score corrected by colony size

Human disturbance including rapid urbanization and increased temperatures can have profound effects on the ecology of local populations. Eusocial insects, such as ants, have adapted to stressors of increasing temperature and urbanization, however these evolutionary responses are not consistent among populations across geographic space. Here we asked how urbanization and incubation temperature influence critical thermal maximum (CTmax) and various ecologically relevant behaviors in three ant species in urban and rural locations in Worcester, MA, USA. We did this by incubating colonies of three species of cavity dwelling ant (Aphaenogaster picea, Tapinoma sessile, and Temnothorax longispinosus) from 2 habitat types (Rural and Urban), for 60-days at multiple temperatures. We found that incubation temperature, urbanization and species of ant all significantly affected overall colony critical thermal maximum. We also found that recruitment time, colonization time and defense response were significantly affected by incubation temperature and varied between species of ant, while recruitment and colonization time were additionally affected by urbanization. These variable changes in performance and competitive traits across species suggest that responses to urbanization and shifting temperatures are not universal across species. Changes in behavioral responses caused by urbanization may disrupt biodiversity, creating unusual competitive environments as a consequence of natural adaptations and cause both direct and indirect mechanisms for which human disturbance can lead to local species extinction.