Analysis, classification and identification of gastropod locomotive mucus by Fourier transform infrared spectroscopy

Mucus is a defining feature of many gastropod phenotypes. Its material properties are a product of selection acting on the relationship between mucus composition, structure and performance; however, to date, there are very few studies addressing this. Here, we introduce attenuated total reflection Fourier transform infrared (FTIR) spectroscopy as an accessible analytical technique to record, identify and classify gastropod locomotive mucus, as deposited, with practically no intervention. Advancing the field beyond previous spectroscopic studies, we also probe mucus response to a temperature increase, determining its propensity to remain hydrated (i.e. resist denaturation) and serving as the basis for building cladograms based on spectral similarities. Specifically, the relative content of proteins, their secondary structures, protein-carbohydrate interactions, and thermal stability were investigated and shown to have significant variation between species. Collating and comparing over 648 spectra across 12 species and 7 superfamilies, we report that it is even possible to classify gastropods based solely on their secreted pedal mucus. This was further compared to previously published phylogenetic data and taxonomies enabling a 100% hit rate of genus assignment, implying that mucus FTIR may also be a powerful tool for field-based biodiversity studies.