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Difficulty in observing oestrus is a problem for many dairy farmers performing AI. Finding ways to synchronize oestrous cycles or strengthen display of oestrus without hormonal treatments would be of great interest because many consumers object to the use of exogenous hormones on healthy animals. Modification of reproductive cycles through chemical communication has been reported in several species including cattle. LH is an important regulator of the follicular phase and could possibly be subject to pheromonal influence. This study focuses on the effect of volatile compounds from oestrous substances on LH pulsatility preceding the preovulatory LH surge in cattle. Four heifers of the Swedish Red breed were kept individually in isolation. Exposure to water during the control cycle (CC), and bovine oestrous urine and vaginal mucus during the treated cycle (TC), started simultaneously with induction of oestrus. Blood sampling at 15-min intervals started 37 h after administration of PGF and continued for 8 h. Monitoring of reproductive hormones, visual oestrus detection and ultrasonographic examination of the ovaries continued until ovulation had occurred. The mean concentration of LH at pulse nadir was significantly higher during TC (2.04 ± 0.18 ng/ml) than during CC (1.79 ± 0.16 ng/ml), and peak amplitude was significantly higher during CC (Δ1.03 ± 0.09) than during TC (Δ0.87 ± 0.09). No other parameters differed significantly between the two cycles. We conclude that the difference in LH pulsatility pattern may be an effect of exposing heifers to oestrous vaginal mucus and/or urine and that the mechanism behind this needs further investigation.