Urbanisation and anthropogenic habitat destruction are pervasive threats to wildlife, as extensive landscape modification creates novel environments in which only a subset of species are able to survive. Behaviour is a significant factor in the success of these urban exploiters, as animals with certain behavioural responses may be more adept at dealing with novel habitats and species interactions.  We assessed the boldness and aggression of the European Garden spider, Araneus diadematus, along an urban gradient in Hamburg, Germany.  Habitat and anthropogenic factors around each spider were quantified at local (vegetation complexity and land cover over a 10m radius) and landscape (socio economic and spatial data) scales and combined into an index of urbanisation. The behaviour response variables (response to a simulated predator attack, activity in a novel environment and superfluous killing of prey) were combined into three indices representing aggression, response to threat and boldness. Although urbanisation has previously been assumed to favour bold, exploratory individuals, we found that rural spiders were significantly more aggressive and sub urban spiders were the most active. This is possibly a result of higher densities of spiders in urban habitats and reflects the fact that behavioural responses to urbanisation are not universal across taxa. As a result, we need a better understanding of the effects of anthropogenic disturbance on a broad range of taxa in order to manage biodiversity in urban systems.