A quantitative analysis of objective feather colour assessment: measurements in the lab are more reliable than in the field

The evolution of animal colouration is importantly driven by sexual selection operating on traits used to transmit information to rivals and potential mates, which therefore, have major impacts on fitness. Reflectance spectrometry has become a standard colour-measuring tool, especially after the discovery of tetrachromacy in birds and their ability to detect UV. Birds? plumage patterns may be invisible to humans, necessitating a reliable and objective way of assessing colouration not dependent on human vision. Plumage colouration measurements can be taken directly on live birds in the field or in the lab (e.g. on collected feathers). Therefore, it is essential to determine which sampling method yields more repeatable and reliable measures, and which of the available quantitative approaches best assess the repeatability of these measures. Using a spectrophotometer, we measured melanin-based colouration in barn swallows? (Hirundo rustica) plumage. We assessed the repeatability of measures obtained with both traditional sampling methods separately to quantitatively determine their reliability. We used the ANOVA-based method for calculating the repeatability of measurements from two years separately, and the GLMM-based method to calculate overall adjusted repeatabilities for both years. We repeated the assessment for the whole reflectance spectrum range and only the human-visible part, to assess the influence of the UV component on the reliabilities of sampling methodologies. Our results reveal very high repeatability for lab measurements and a lower, still moderate to high repeatability, for field measurements. Both increased when limited to only the human-visible part, for all plumage patches except the throat, where we observed the opposite trend. Repeatability between sampling methods was quite low including the whole spectrum, but moderate including only the human-visible part. Our results suggest higher reliability for measurements in the lab and higher power and accuracy of the GLMM-based method. They also suggest UV reflectance differences amongst different plumage patches.