The aim of the study was to determine the effect of time after mating on the capacitation status of spermatozoa and the distribution of capacitated spermatozoa in the reproductive tract of Merino ewes. Thirty five ewes were examined for capacitation status in the female reproductive tract at 3, 6 and 24 hours after mating. Samples of fluid and mucus from various sites in the reproductive tract of ewes were collected into HSOF medium for determination of capacitation status. The chlortetracycline-fluorescence assay was used to assess the capacitation status of spermatozoa. Analysis of variance one way classification was used to determine the effect of time after mating on the capacitation status. The mean percentage of capacitated spermatozoa at 3 hours after mating in the anterior vagina was 29.5% and 27.3% in the posterior cervix increasing to about 80% in the anterior uterus. At 6 hours after mating, between about 90% of spermatozoa in the vagina were capacitated and about 75% of spermatozoa in the cervix and uterus were capacitated whereas the percentage of capacitated spermatozoa in the oviducts was up to about 87.5% in the isthmus and 93.5% in the ampulla. All of the capacitated spermatozoa in the oviducts at 6 hours after mating were acrosome-reacted and none had an intact acrosome. By 24 hours after mating, all or almost all spermatozoa were capacitated and acrosome-reacted. Significantly more capacitated acrosome-reacted spermatozoa were present in the ipsilateral oviduct and uterine horns than the contralateral side but there was no significant effect on the percentage of capacitated acrosome-intact spermatozoa
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