Tephritid mass-rearing systems require an artificial substrate for pupation. Pupation substrate characteristics influence the quality of insects produced. Coconut fiber, as an alternative to the conventional pupation substrate vermiculite, was evaluated for Anastrepha obliqua Macquart (Diptera: Tephritidae) pupation behavior (pupation patterns, distribution, respiration rate, and pupal weight) and adult fitness (adult eclosion time, flight ability, and male mating competitiveness). Pupation percentage at 24 h, pupal weight, and flight ability were not significantly affected by substrate type. Adult eclosion levels of 50% were reached at 29.7 and 41.6 h for coconut fiber and vermiculite, respectively. Pupae distribution patterns differed between substrates because the larval aggregation level was reduced during the pupation process in coconut fiber. The pupae aggregation was three times greater in vermiculite than in coconut fiber. A higher respiratory rate in the last days of pupation and adult eclosion were recorded in the insects maintained in coconut fiber. Coconut fiber suitability as a pupation substrate for quality mass production of pupae and its implications for sterile insect technique are discussed.