Background: Animal temperament is de ned as the consistent behavioural and physiological differences that are seen between individuals in response to the same stressor. Neurotransmitter systems, like serotonin and oxytocin in the central nervous system, underlie the variation in temperament in humans. The variations like single nucleotide polymorphisms (SNPs) in the genes for tryptophan 5-hydroxylase (TPH2), the serotonin transporter (SLC6A4), the serotonin receptor (HTR2A), or the oxytocin receptor (OXTR) are associated with the behavior phenotypes in human. Thus, the objective of this study was to identify SNPs in TPH2, SLC6A4, HTR2A and OXTR and to test if those variations predict the temperament of Merino sheep. Results: Using ewes from the UWA temperament ock, that has been selected on emotional reactivity for more than 20 generations, eight SNPs (rs107856757, rs107856818, rs107856856 and rs107857156 in TPH2, rs20917091 in SLC6A4, rs17196799 and rs17193181 in HTR2A, and rs17664565 in OXTR) were found to be distributed differently between calm and nervous sheep. These eight SNPs were then genotyped in 260 sheep from a non-selected ock that has never been selected on emotion reactivity, followed by the estimation of the behavioural traits of those 260 sheep using an arena test and an isolation box test. We found that several SNPs in TPH2 (rs107856757, rs107856818, rs107856856 and rs107857156) were in strong linkage disequilibrium, and all were associated with behavioural phenotype in the non-selected sheep. Similarly, rs17196799 in HTR2A was also associated with the behavioural phenotype. Conclusions: We thus conclude that, rs107856856 and rs17196799 could be used as gene markers for the temperament of Merino sheep, with allele C of rs107856856 and allele A of rs17196799 being associated with calm temperament. Background Animal temperament is often referred to as personality or a behavioural syndrome [1]. More speci cally, animal temperament is the consistent behavioural and physiological differences that are observed between individuals in response to an eliciting event [2]. The variability in temperament is often described along dipoles such as shy to bold, sociable to aggressive, exploring to avoiding, or behavioural activity or inactivity [2]. That individual variability is thought to re ect the individual's perception of the surrounding situation. Non-human animals, in a similar way as humans, evaluate a situation based on particular characteristics of the eliciting event such as its suddenness, familiarity, pleasantness, controllability, and predictability, and how the event accords or deviates from their expectations [3]. The perception of a situation leads to an emotional state that is experienced by the individual. The emotion then elicits physiological and behavioural responses, and those responses have been used to qualify the temperament of individual animals [3, 4]. In sheep, the phenotype and the genetic heritability of temperament traits [5-7], have been assessed using behavioural tests. Tempera...