The effect of buffalo follicular fluid (buFF) on follicular development, estrus response and luteal function was investigated in anoestrous does. Treatment with buFF (18 ml/doe) had no significant effect on the number of antral follicles of all class categories during the period of administration. However, after cessation of buFF treatment, the number of total antral follicles increased significantly with time ( P < 0.003) as well as due to the treatment × time interaction ( P < 0.02), without any influence on follicle size. Injection of buFF also caused a marked increase ( P < 0.049) with time in the number of medium-sized follicles at cessation. Approximately 60 and 20% of buFF-treated anoestrous does showed behavioural and silent estrus, respectively, compared to none in the control. The mean interval between cessation of buFF treatment to onset of oestrus and oestrus duration was 67.0 ± 18.5 and 17.0 ± 3.6 h, respectively. Corpus lutea size varied between 4.6 and 5.8 mm with an average diameter of 5.2 ± 0.3 mm. Only 33.3% of does showed serum progesterone levels above 1 ng/ml, while the remainder (66.7%) had below 0.5 ng/ml. Our results indicate that exogenous administration of buFF causes enhanced follicular activity following cessation of treatment, which results in behavioural oestrus and corpus luteum (CL) development in anoestrous does. CL development and its function is, however, inadequate in buFF-treated anoestrous does.