Lidocaine is used for epidural and spinal anesthesia in various animal species. The ideal drug for epidural and spinal anesthesia should have a long effective duration in addition to a fast onset of action, and adequate analgesia and muscle relaxation. Despite the delayed onset of action, bupivacaine provides a longer duration of anesthesia than lidocaine. The purpose of this study was to compare the onset to effect and duration of action between lidocaine and bupivacaine for spinal anesthesia in broiler chickens. Thirty-two, 8-week-old, female Ross broiler chickens were randomly divided into 4 groups of 8: 1) 2 mg/kg lidocaine (L); 2) 0.1 mg/kg bupivacaine (B0.1); 3) 0.25 mg/kg bupivacaine (B0.25); and 4) 0.5 mg/kg bupivacaine (B0.5). After aseptic preparation, a 23-gauge spinal needle was inserted into the synsacrococcygeal space of the chickens with correct needle placement confirmed by a sudden loss of resistance. Spinal anesthesia was performed with the aforementioned doses of lidocaine and bupivacaine. The respiratory rate and cloacal temperature were measured every 10 minutes in each chicken until the anesthetic effect was no longer present. The onset to effect and the duration of action were calculated for each bird based on the pinch test at predetermined time intervals. The results are demonstrated as mean ± SD. The onset of action for bupivacaine (9 ± 1.41, 4.33 ± 1.15, and 3.33 ± 1.23 minutes in B0.1, B0.25, and B0.5 groups, respectively) was significantly delayed compared with that of lidocaine (1.37 ± 0.52 minutes). The duration of action of B0.5 (54 ± 6.08 minutes) was significantly longer than that of any other group (17.87 ± 3.18, 11 ± 1.41, and 18 ± 4.36 min in L, B0.1, and B0.25 groups, respectively). The results showed that a spinal injection of 0.5 mg/kg bupivacaine produces approximately 55 minutes of spinal anesthesia in these broiler chickens, which is much longer than the 18 minutes of anesthesia provided by 2 mg/kg lidocaine. Considering the various disease conditions that affect the cloacal area of birds, one can use each of these anesthetic drugs for either short-term or long-term spinal anesthesia in chickens and possibly other avian species.