The laboratory biology of Ornithodoros (Alectorobius) puertoricensis Fox was studied over a 2-yr period. Approximately 100-150 ticks were reared individually at each of four temperatures: 22, 27, 33, and 40 degrees C and 90-95% RH. The mean egg incubation periods at those temperatures were 20.3, 11.1, 7.3, and 6.1 d, respectively. The average larval feeding period was 5.8 +/- 1.5 d for 15,875 larvae that fed on guinea pigs. The development times for first to fourth nymphal instars were as follows: 11.7, 48.5, 75.1, and 92.1 d, respectively, at 22 degrees C; 5.8, 18.8, 38.0, and 36.0 d, respectively, at 27 degrees C; 4.2, 10.5, 14.9, and 38.1 d, respectively, at 33 degrees C; and 5.8, 10.7, 21.2, and 35.3 d, respectively, at 40 degrees C. Males usually eclosed after three or four molts, and females usually eclosed after four or five molts. Approximately 10% of all nymphs required more than one blood meal per instar at least once during development. Twenty pairs of adults were held at each of three temperatures (22, 27, and 33 degrees C) for a year to study reproductive behavior. The number of gonotrophic cycles per female per year was 6.9, 9.8, and 10.8 at 22, 27, and 33 degrees C, respectively. The mean duration of the gonotrophic cycle was 42.3 d at 22 degrees C, 25.5 d at 27 degrees C, and 20.5 d at 33 degrees C. Mean egg production per female per gonotrophic cycle was 151 at 22 degrees C, 117 at 27 degrees C, and 130 at 33 degrees C and was not affected by temperature. O. puertoricensis did not exhibit autogeny or parthenogenesis. Hyperparasitism was observed in immatures and adults.