Transovarial transmission experiments were conducted with three groups of Ornithodoros (Pavlovskyella) marocanus Velu; one group consisted of 27 pairs of adults that had been fed as larvae on a pig with a viremia of 10(7.4) HAd50/ml of African swine fever virus (ASFV). The second and third groups each consisted of 100 pairs of adults fed on a viremic pig (10(4.5) HAd50/ml) as adults. The first group underwent five gonotrophic cycles over a 554-d period. The second and third groups underwent three and two gonotrophic cycles, respectively. All larvae were fed in individual cohorts on naive pigs and the resulting nymphs were assayed by cohort for ASFV. None of the larvae transmitted ASFV to naive pigs by bite and ASFV was not isolated in swine buffy coat cultures from any cohort of nymphs. Therefore, O. marocanus does not exhibit transovarial transmission of ASFV. Venereal transmission experiments were conducted with pairs of O. marocanus in which either the female (100 pairs) or the male (100 pairs), respectively, had fed on a viremic pig (10(4.5) HAd50/ml). Both groups underwent at least two gonotrophic cycles over a 470-d period, were sampled periodically for the presence of ASFV, and the progeny were tested for the presence of ASFV. Venereal transmission from male to female occurred in 10% (1/10) of O. marocanus after the first gonotrophic cycle, but not after the second or third gonotrophic cycle, and transovarian transmission in these groups was not observed. Venereal transmission from infected females to uninfected males did not occur. ASFV persisted through five gonotrophic cycles over a 554-d period in 30% of adults fed on a viremic pig as larvae. ASFV was cleared during three gonotrophic cycles within a year from nearly all ticks fed on a viremic pig as adults. Virus-induced mortality rats of 12-80% occurred among ticks fed on viremic animals, whereas, no mortality was seen in ticks fed on uninfected animals. ASFV infection in ticks did not effect feeding frequency, egg-hatch rate, or the oviposition rate among females fed on a viremic pig as adults. The oviposition rate for females fed on a viremic pig as larvae was reduced by 63.4%. Parthenogenesis was not observed among O. marocanus. The mean gonotrophic cycle duration for pig-fed O. marocanus at 27 degree C was 19.8 d and the mean fecundity was 88.3 +/- 17.9 eggs/female/gonotrophic cycle.