This study focuses on anti-trauma mats designed for buffaloes’ comfort, using as raw materials rubber powder from end-of-life tyres (ELTs) and an isocyanate-based polyurethane resin binder. The first part of the study focused on mat formulation. Whilst it was possible to select a unique combination of raw materials and design features, it was necessary to investigate the relationship between three critical parameters affecting mat consistency and therefore buffalo comfort: binder quantity, mat thickness, and desired final mat density (bulk). In order to quantitatively assess the variation in hardness, various combinations were investigated within well-defined ranges based on the relevant literature. The results obtained from nine selected combinations indicate that increases in the three critical parameters do not induce a real phase transition in the final product consistency, although the hardness suggests an increasing trend. The mats consistently exhibited a moderately soft/hard consistency, offering environmental benefits in terms of increased rubber usage and potentially reduced chemical binder, depending on the desired thickness. The selected mixture showed excellent resistance to heavy chemical loads, suggesting reliability for frequent cleaning operations. The second part of the study involved field trials of the mats with calves. This involved monitoring their weight gain and appetite levels over a 90-day period. The results showed excellent growth performance compared to uncoated grids (i.e., weight gain was approximately 20% higher at the end of the observation period); this was similar to that achieved with the use of straw bedding. However, compared to straw bedding, the mats (i) exhibit long-term durability, with no signs of wear from washing or trampling over the months of the trial, (ii) allow for quick and efficient cleaning, and (iii) enable companies to save on labour, material (straw), and waste disposal costs, while maintaining (or even improving) the same welfare levels associated with the use of straw.