Humans represent numbers along a mental number line (MNL), where smaller values are located on the left and larger on the right. The origin of the MNL and its connections with cultural experience are unclear: Pre-verbal infants and nonhuman species master a variety of numerical abilities, supporting the existence of evolutionary ancient precursor systems. In our experiments, 3-day-old domestic chicks, once familiarized with a target number (5), spontaneously associated a smaller number (2) with the left space and a larger number (8) with the right space. The same number (8), though, was associated with the left space when the target number was 20. Similarly to humans, chicks associate smaller numbers with the left space and larger numbers with the right space.