A considerable number of studies have reported differences among dog breeds with respect to their genetic profile, cognitive abilities or personality traits. Each dog breed is normally treated as a homogeneous group, however, researchers have recently questioned whether the behavioural profile of modern breeds still reflects their historical function or if the intense divergent selective pressures and geographical barriers have created a more fragmented picture. The majority of studies attempting to assess and compare modern breeds' personality focused on the evaluation of adult dogs where the potential effects of environmental/human factors on the dogs' behaviour are hard to discern from their genetic heritage. In the following study, we aimed at investigating between-and within-breed differences in the personality of two-months-old puppies by direct behavioural observation of 377 puppies from 12 breeds. Results showed that there was no effect of sex, however both breed and litter, significantly affected all personality traits. Breed on average explained 10% of the variance, whereas the effect of litter was noticeably higher, explaining on average 23% of the variance. Taken together, our results suggest that breed does have some influence on personality traits, but they also highlight the importance of taking litter effects into account.The assessment of dogs' personality has gained increasing attention in the last decades for its potential applicability 1-4 , as well as for more theoretical aspects, such as the genetic basis of complex behaviour 5,6 . One of the topics, strictly linked to dog personality that is still being debated, is whether dogs from different breeds significantly diverge on specific behavioural traits, potentially reflecting their historical function (original selection to optimise their performance in specific tasks as stated by the kennel clubs standards) 7-9 . Breed profiling has largely been based on historical and anecdotal notions rather than scientific evidence 8 . However, more recently scientific attempts to provide reliable profiling of modern breeds, taking into account the large within-breed variability due to different selective pressures and geographical barriers, have been carried out 7,8,[10][11][12] . Svartberg 7 , for example, found that within the same breeds, irrespective of their historical function, individuals in lines selected for companionship showed high levels of playfulness, whereas selection for use in dog shows correlated positively with fearfulness and negatively with playfulness, aggressiveness, and curiosity. However, thus far, the majority of studies attempting to assess breed personality have been carried out using questionnaire-based methods 8,13,14 and have largely focused on the evaluation of adult dogs 7, 9 . Yet, behavioural variability towards different stimuli and situations can be detected also in puppies [15][16][17][18] and arguably, if there are breed differences in temperament, observing these in puppies would provide stronger evidence, since ...