Abstract: Horses are likely to exhibit aggression when meeting for the first time. Therefore, this study compared 3 methods for mixing horses to evaluate their effectiveness in reducing aggressive interactions: (1) mixing pairs of horses in a paddock (P, 10 minutes, 15 tests), (2) introducing 1 unfamiliar horse to a pair of familiar, resident horses in a paddock (PP, 10 minutes, 15 tests), (3) allowing limited physical contact between pairs of horses for a short period of pre-exposure in neighboring boxes (B, 5 minutes, 16 tests) before mixing them in a paddock (BP, 10 minutes 16 tests). A total of 16 Swedish Standardbred mares, aged 6-18 years (mean age ± SD: 11 ± 4.4), were included in the study. Half of the horses were familiar with each other (resident horses, n = 8), whereas the other half were bought in from a variety of sources (unfamiliar horses, n = 8). Social interactions, consisting of behaviors from the sender, the receiver, and the subsequent sender's response, were recorded continuously as frequencies. There were no differences in the frequencies of aggressive behaviors between the 3 mixing methods, including those aggressive behaviors in which physical contact had been attempted (kick, strike). Although resident horses were overall more aggressive (median number of aggressive behaviors per horse, 62; Q1, 36; Q3, 68.5) than unfamiliar horses (median per horse, 4; Q1, 2; Q3, 12.5) during all tests (U = 97, P = 0.003), none of the 62 tests needed to be terminated. Unfamiliar horses did not receive more aggression from resident horses in PP (mean per test ± SD: 5.1 ± 3.1) than in P (mean per test ± SD: 6.4 ± 4.9) (t = 0.63, P = 0.544). However, the behavior “attack” was more frequent in PP (median per test, 2; Q1, 0; Q3, 5) than in P (median per test, 0; Q1, 0; Q3, 1) (U = 282, P = 0.042), and “flee” was more frequent in PP (median per test, 6; Q1, 4; Q3, 8) than in P (median per test, 1; Q1, 0; Q3, 6) (U = 290, P = 0.018). Pre-exposure in boxes did not reduce aggression in BP (median per test, 7; Q1, 4.3; Q3, 11.8) as compared with P (median per test, 6; Q1, 2; Q3, 16) (U = 264, P = 0.767), but during pre-exposure in B tests, horses exchanged more nonaggressive (median per test, 2; Q1, 0.3; Q3, 4) than aggressive (median frequency of aggressive behavior, 0; Q1, 0; Q3, 1) (W = 71, P = 0.013) and mixed interactions (median per test, 0; Q1, 0; Q3, 1) (W = 92, P = 0.016) through the opening. Results suggest mixing an unfamiliar horse with 2 resident horses at the same time instead of one by one may be preferable. In this way, the total aggression received by the unfamiliar horse will potentially be less, even though aggressive interactions may be more intense.