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The objective of the present study was to characterize the variation in oestrous behaviour among ovariectomized cows in response to a measured dose of oestradiol benzoate (OB) . In study 1, nine ovariectomized Galloway cows, approximately 10 years old, were challenged with an intramuscular injection of either 0·25, 0·5 or 1·0 mg of OB. Following this, 0·5 mg OB was chosen as the appropriate dose required to induce oestrous behaviour in ovariectomized Galloway cows. In study 2, nine cows injected with 0·5 mg OB were monitored for oestrous behaviour using KAMAR® heat mount detectors. Blood samples for plasma oestradiol-17β (E2) assay were taken every 4 h between 0 to 72 h and once at 96 h, following OB administration. The plasma E2 concentrations between 0 to 96 h following OB administration differed significantly ( P < 0·001) among cows. The interval to peak E2 concentrations averaged 17·42 (s.e. 2·21) h and the mean peak E2 concentration was 5·86 (s.e. 0·57) ng/l. The mean interval from OB administration until onset of mounting activity was 24·57 (s.e.2·38) h and the duration of oestrus averaged 10·5 (s.e. 1·99) h . In all cows, mounting activity occurred following peak E2 concentrations after an average interval of 7·73 (s.e. 1·84) h. There was no significant association between peak E2 concentrations and the interval to onset of mounting activity or its duration. Since the variation among Galloway cows in the manifestation of behavioural oestrus was independent of systemic E2 concentrations, it implies that there are inherent differences between individuals in the sensitivity of the hypothalamus to physiological E2 thresholds. This may help to explain the incidence of silent ovulation in some animals and the occurrence of overt oestrus associated with follicular activity during the luteal phase of the cycle or during the various stages of pregnancy, in the presence of high plasma progesterone concentrations.