The goal of this research was to evaluate milking temperament and its relationship with test-day milk (TDMY0) yield in Bulgarian Murrah buffaloes. This study involved 90 buffalo cows reared under a tie-stall production system which were milked twice a day with a milking pipeline. The behavioral responses of the buffaloes were reported during preparation for milking and during actual milking. The average temperament score during preparation for milking was 1.83, and 1.93 during milking itself. The most common reaction was leg lifting (18.9%), followed by cows moving on the stall bed (10%), definite kicking (9.9%), and 13.3% managing to remove the milking cluster during milking. The frequency of buffaloes showing adverse reactions (scores 4 and 5) increased considerably during milking compared to preparation for milking. Repeated scoring of temperament during the same lactation did not show a significant difference in the frequency of temperament assessments both in preparation for milking and during milking. The minimal difference may be due to the accuracy of the assessment or a momentary change in the condition of the animals during the two scorings. Cows with the most unwanted milking behavior (scores 5 and 4) had the highest LS means for TDMY, 8.18 kg and 7.65 kg, respectively. The reasons for these buffaloes remaining until later lactations was their high milk yield and the injection of oxytocin before milking, which helps them to be fully milked.