This study investigated the hypothesis that a reduction in submission rates at a resynchronised oestrus is not due to the resynchrony treatment involving intravaginal progesterone releasing devices (IVDs) and oestradiol benzoate (ODB) but is associated with artificial insemination (AI) at the first synchronised oestrus. In Experiment 1, cows were synchronised for first oestrus with IVDs, with ODB administered at the time of device insertion (Day 0, 2 mg IM) and 24 h after removal (Day 9, 1 mg IM) and PGF(2alpha) injected at the time of device removal. Cows were then either inseminated (I) for 4 days or not inseminated (NI) following detection of oestrus (first round of AI). Every animal was resynchronised for a second round of AI by reinsertion of IVDs on Day 23 with administration of ODB (1 mg IM) at the time of insertion as well as 24 h after removal (Day 32). Cows detected in oestrus and inseminated for 4 days at the second round of AI were resynchronised for a third round by repeating the resynchrony treatment starting on Day 46 and inseminating cows on detection of oestrus for 4 days. In Experiment 2 the same oestrous synchronisation and resynchronisation treatments were used, but the timing of treatments differed. The cows had their cycles either presynchronised (treatment start Day -23) without AI and then resynchronised, starting on Day 0, for the first round of AI for AI at detected oestrus for 4 days, or they were synchronised (treatment start Day 0) for the first round of AI. In Experiment 1, 91.4% (64/70) and 92.6% (63/68) (P = 0.79) of cows in the I and NI treatments, respectively, were detected in oestrus after the initial synchronisation. At the second round of AI, submission rates for insemination were lower in the I group compared to the NI cows (74.5%, 35/47 versus 92.6%, 63/68, respectively; P = 0.007). Pregnancy rates (proportion treated that were classified as becoming pregnant) in I and NI cows 4 weeks (61.4%, 43/70 versus 63.2%, 43/68) and 7 weeks (77.1%, 54/70 versus 69.1%, 47/68) after the AI start date (AISD) did not differ significantly between treatments. In Experiment 2, presynchronisation and then resynchronisation of oestrous cycles before the first round of AI did not affect oestrous detection rates at the first round of AI (100%, 44/44 versus 98.0%, 50/51; P = 0.54), or pregnancy rates 1 week (63.6%, 28/44 versus 60.8%, 31/51; P = 0.70), 4 weeks (72.7%, 32/44 versus 76.5%, 39/51; P = 0.76) and 7 weeks (81.8%, 36/44 versus 88.2%, 45/51; P = 0.40) after AISD compared to cows that had their cycles synchronised for the first round of AI. These findings support our hypothesis that a reduction in submission rates at a resynchronised oestrus is associated with AI at the first synchronised oestrus and not due to a resynchrony treatment involving IVDs and ODB. This study supports the concept that early embryonic loss following AI at a synchronised oestrus could cause a reduction in submission rates following resynchronisation of oestrus, although investigation of the effect of passing an AI catheter or semen components were not studied per se.