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The present study was undertaken to test the effect of two vulva injections of D-cloprostenol on day 7, 9 and 10 of oestrous cycle on the duration of the interestrous interval in gilts. Following a pre-treatment oestrous cycle, 87 gilts were assigned to receive vulva injections of 75 μg D-cloprostenol at 08:00 and 14:00 h on day 7 (D7; n=30), day 9 (D9; n=29) or day 10 (D10; n=28) of their second observed oestrous cycle. Across the treatments, the duration of the oestrous cycle with D-cloprostenol treatment (19.1±0.1 d) was not different from that of the previous oestrous cycle (20.1±0.4 days). Plasma progesterone concentrations were evaluated 6 h before and 24 and 72 h after D-cloprostenol treatment in the D9 group. Compared to pretreatment levels (9.6±0.4 ng/mL), plasma progesterone concentrations were reduced (P<0.05) at 24 h (6.3±1.0 ng/mL) and 72 h after treatment but complete luteolysis did not occur. These data indicate that in gilts double vulva administration of D-cloprostenol is not able to induce a complete luteolisys and hence the duration of the oestrous cycle is not modified.