Gastropods use adhesive locomotion to successfully traverse a range of substrates by adapting the properties of a thin mucus layer. Sample collection methods can disrupt the native mucus structure, so to investigate this system effectively a method for visualising the native deposition of gastropod mucus trails in situ was developed. Optical profilometry and video analysis were utilised to uncover how animal behaviour impacts mucus structure and analysis of the dried trail topography provided insight into mucus alignment and deposition rate.

Our results revealed that dried gastropod trails have a non-uniform deposition profile with ridges visible along both edges of the trail. This phenomenon was independent of specimen size for the slug species Arion vulgaris, Arion flagellus and Limax maximus, but direction and velocity did impact the prominence of the trail ridges. The dried trail topography was the inverse of the convex deposition profile observed in the rehydrated mucus trails of a marine gastropod, indicating a potential difference in mucin deposition or alignment that is independent of the mucus hydration state. When compared to the model slug species, Ariolimax columbianus, the species from this study had thinner dried mucus trails, highlighting potential variations in the cost of adhesive locomotion between species. Our results suggest that mucus deposition rate and animal behaviour have a greater impact on mucus trail topography than species or specimen mass. Moreover, analysis of the dried mucus trails can provide invaluable insight into the relationship between the hydration and mucin costs associated with mucus excretion and adhesive locomotion.