*2.1. Tympanic Temperature (TT)*

1

TT was used as an indicator of body temperature. Data logger devices (ibuttons, Maxim Integrated Products Inc., San Jose, CA, USA) were manually installed into the tympanic canal using the same procedure described by Arias et al. [25]. Devices were fitted 24 h before initiating the experiment. Data loggers registered TT every 10 min; thus, in order to obtain values for the sample times (before loading, after unloading), the measurements from the 3 h before and after transport were averaged for each calf. The data from two calves and all TT records after the 24 h fasting period were omitted from the analysis as anomalous values were obtained, probably because of device displacement inside the calves' ears.

#### *2.2. Maximum Eye Temperature (MET)*

MET was obtained by capturing infrared images of both eyes at approximately 0.3 m distance and a 90◦ angle from the individual using a thermal camera (FLIR i5, FLIR Systems, Wilsonville, OR, USA), calibrated with an emissivity(ε) of 0.95, according to information provided by the manufacturer. Image analysis was performed using the software FLIR Tools 3.1 (FLIR Systems, Wilsonville, OR, USA) and atmospheric temperature and relative humidity were included in the calculation. The image obtained was used for analysis, as shown in Figure 1. The MET of both eyes was averaged to obtain one MET value for each calf at each sampling.

**Figure 1.** Infrared image of the eye region of a calf. Red triangle indicates maximum eye temperature (MET).
