DOI: 10.15482/usda.adc/28459058.v1
title: Data from: Temporal plasticity of thermal tolerance in ants

description: Three .csv files. Two years of data collected on critical thermal maxima and minima (CTmax and CTmin) from 14 colonies in Oklahoma from 2017-2018. Additional data files represent thermal traits from 10 colonies in a short term (10 days) acclimation experiment and projections using environmental data for potential activity differences.<b>Abstract from paper:</b>How do individuals tolerate both the hot and the cold climate of our planet? One possibility is that organisms have plastic traits like thermal tolerance that allow them to function in highly variable environments. In this study, we tested whether phenotypic plasticity of temperature tolerance (i.e. acclimatization in the field and acclimation in the lab) occurs in the red harvester ant, <i>Pogonomyrmex barbatus</i>, at two temporal scales. We first measured the upper and lower critical thermal limits (CT<sub>max</sub> and CT<sub>min</sub>) of ants monthly for two years while concurrently measuring environmental conditions. Both CT<sub>max</sub> and CT<sub>min</sub> co-varied with temperature in a predictable way; values increased in a positive, linear manner. We then experimentally tested whether CT<sub>max</sub> and CT<sub>min</sub> could shift within a shorter time period by exposing subcolonies of ants to cool (10°C), moderate (20°C), and hot (30°C) temperatures for 10 days. CT<sub>max</sub> increased only slightly at the hottest temperature treatment (+1.2°C), however CT<sub>min</sub> increased considerably under both moderate (+2.6°C) and hot treatments (+3.8°C). Combined, our results suggest that thermal tolerance of ants may be more plastic than originally hypothesized, potentially aiding an already thermophilic clade.<b>Methods from paper:</b><i>Study site and environmental temperature</i>We sampled ant workers monthly during their annual active period in 2017 and 2018 (i.e. March-November) from 14 colonies in a 30-ha grazed prairie in the central Great Plains of Oklahoma (34.5478º N, -98.2311º W, 330 m elevation). Over two years, ground temperature was recorded every 10 minutes using HOBO U23 Pro v2 External Temperature Data loggers at three equidistant locations within the sampling area. Temperature values were then averaged per month.<br><i>Thermal tolerance across months</i>During each sampling event (n = 18), we collected ~20 workers directly outside the nest of each colony and used 5 workers to measure critical thermal maximum (CT<sub>max</sub>) and 5 workers to measure critical thermal minimum (CT<sub>min</sub>). We did so using a heating/cooling assay to determine the temperature at which individuals lost muscle control. Thermal assays were conducted by placing individual ants into 1.5ml microcentrifuge tubes and plugging the tops with cotton to remove a potential thermal refuge in the cap. For CT<sub>max</sub>, tubes were placed randomly into a Thermal-Lok 2-position dry heat bath that was prewarmed to 36ºC. Every 10 minutes, individuals were checked to see if they had reached their critical thermal limit by rotating the tube to check for a righting response. The temperature was then increased by 2ºC, with the process repeated until all individuals had reached their critical thermal maximum. CT<sub>min</sub> was assayed in a similar manner, but we used a EchoThermTM IC20 chilling/heating dry bath that was precooled to 20ºC, following the methods above except with temperature lowered 2°C every 10 minutes. Additional ants from each colony were kept at ambient conditions as a control during each thermal assay, all of which survived. During each trial, we also confirmed the interior temperature of one unused vial using a thermocouple attached to an Extech MN35 Digital Mini MultiMeter. CT<sub>max</sub> and CT<sub>min</sub> values were averaged per colony for each month.<br><i>Thermal tolerance within a month</i>In April of 2019, we collected ~200 workers from each of 10 separate colonies to assess if critical thermal limits could change within a single cohort of ants over a short period of time. We split each group of 200 workers into three sub-colonies containing 50 workers and placed these newly created sub-colonies into three environmental chambers set at 10ºC, 20ºC, and 30ºC with a 12:12 L:D cycle and 85% RH. The selected temperatures span the approximate range of average monthly temperatures at our study site during which ants were active. Each sub-colony was provided with water and 20% sucrose solution ad libitum in cotton plugged vials and a small petri dish with Plaster of Paris that was moistened every other day. Critical thermal limits (CT<sub>max</sub> and CT<sub>min</sub>) were assayed using five individuals from each colony immediately prior to the start of the experiment and for five individuals from each sub-colony after 10 days in the environmental chambers. CT<sub>max</sub> and CT<sub>min</sub> values were averaged per colony for each temperature treatment.