Spectra of feather samples are impacted by the substrate color against which they are measured

Many studies of animal and plumage coloration use spectrometry to obtain objective measurements of colorful traits. Objective measurements of plumage color are particularly important as the avian visual system is distinct from our own and subjective color assessment by researchers can miss important color variation or signals. As spectrometry has become one of the preferred methods of color measurement, appropriate methodologies and reporting standards have refined and improved the technique as well as increased our ability to make comparisons across studies. Here, we investigate another important methodological decision by examining how the background color against which feather samples are measured impacts the reflectance spectra and color descriptors commonly extracted from those spectra. We used feather samples collected from Eastern Bluebirds (Sialia sialis) and Barn Swallows (Hirundo rustica erythrogaster) to determine how black and white backgrounds influence color measurements. We measured the same feather sample in the same arrangement against a white index card and a piece of black cardstock and found that measurements taken against a white background tended to yield higher reflectance curves, which also resulted in significant differences in some color descriptors. Lightly colored, wispy, and small feathers seem to be particularly susceptible to background color effects. Thus, researchers using spectrometry to measure colorful traits should carefully consider the background against which they are taking measurements as white backgrounds tend to increase reflectance and certain feather colors or morphology (light and wispy) may be particularly susceptible to spectral contamination. Received 26 May 2022. Accepted 28 December 2022.