The overall objective of this study was to evaluate the effects that milking temperament (MT) of water buffaloes has on milking traits, welfare indicators, and reproductive outputs. The study was performed on 60 multiparous buffalo cows (6.7 ± 0.6 lactations) at the beginning of their lactation (100 days in milk, DIM). Each buffalo cow was scored by two independent observers using a temperament scoring system (1: extremely calm, 2: calm, 3: alert, 4: reactive, and 5: aggressive), and then grouped as ‘calm’ (scores 1, 2, and 3; n = 42) or ‘nervous’ (scores 4 and 5; n = 18). Additionally, the milk yield at 100 DIM (MY), milking speed (MS), calving interval (CI), age at first calving (AFC), body condition score (BCS), animal-based welfare parameters, and infrared thermography data (IRT) were evaluated. The MT significantly influenced the MY (p = 0.0082), with calmer cows outperforming their nervous counterparts. The MS was significantly influenced by the MT (p = 0.0015), with calmer animals having a higher milk ejection rate. The MT of the cows had no influence on the CI, AFC, or BCS. The evidence from this study suggests that the responsiveness of buffalo cows during milking affects their milk yield and milking speed, with no associations being found for reproduction efficiency indicators or animal-based welfare indicators.