We hand-collected adult toads (n = 8 individuals per site) from four sites across the toads’ tropical range within Australia, from Townsville, Qld in the east (GPS coordinates: − 19.26, 146.82, 14 m altitude) to Richmond, Qld (− 20.73, 143.14, 218 m altitude), Middle Point, NT (− 12.56, 131.33, 12 m altitude) and Kununurra, WA (− 15.78, 128.74, 49 m altitude) in the west. That transect spans the toads’ 80-year invasion history. Although both temperatures and precipitation exhibit a general east–west cline, the greatest disparities in the duration of hot dry conditions per year lie between the easternmost site (Townsville) and the three other sites (Fig. 1). We recorded toad mass (after gently squeezing the animal in a standardized manner to induce it to empty its bladder) and snout-vent length (SVL) immediately before conducting the trials.
Data from Australian Bureau of Meteorology7.
Mean climatic conditions in the four sites from which we collected cane toads (Rhinella marina) for use in laboratory trials. The red line connects mean monthly maximum air temperatures, the green line shows mean monthly air temperatures, and the blue line shows mean monthly minimum air temperatures. Histograms show mean monthly rainfall.
Toads were not fed for three days prior to experiments, to ensure they would not defecate during the experiment and minimize variability in mass due to stomach contents. Toads from all four populations were housed in a room kept at 18 °C, then moved concurrently to a temperature-controlled room set at 37 °C. All toads were in separate containers (ventilated plastic boxes of 1-L capacity), half of which had dry paper towel as substrate whereas the other half had 40 mL of water, enough to keep the ventral portion of the body moist but not the rest of the body.
We measured toad body temperatures at the beginning of the trial, and after 20 min and 40 min, using an infrared thermometer (Digitech QM7215) held ~ 10 cm from the toad’s dorsal surface. At the beginning and end of the experiment we measured internal temperatures with a cloacal probe (Digitech QM7215 with probe attachment), to check that our measurements of external body temperature offer robust estimates of internal temperature also. Cloacal temperatures were taken within 10 s of each toad’s removal from the container. After a trial, toads were kept at a temperature of 25 °C, allowed to fully hydrate and monitored for wellbeing during recovery. No adverse effects of the trials were evident.
We used mixed model repeated measures analysis to identify factors affecting body temperatures of cane toads during the 40-min heating trials. Sex and body mass were used as covariates in the analysis with climate at each collection site (# consecutive months per year with average maximum temperature > 30 °C and with < 25 mm rainfall (see above)) and wet/dry treatment as factors and time (0, 20, 40 min) as the repeated measure. Individual ID nested within population was used as a random effect. Our measure of climate characteristics at each site (0, 3, 5, or 6 hot dry months per year) was analysed as a continuous variable. However, for graphical purposes, our figures present data for each site depicted as a categorical variable (by location name). The data conformed to assumptions of normality and variance homogeneity and analyses were run using JMP Pro 14.0 (SAS Institute, Cary, NC).
Ethical approval
All procedures were approved by the University of Sydney Animal Ethics Committee (Protocol #703), and all procedures conformed to Australian guidelines for the care and use of animals during research.
Source: Ecology - nature.com
