Herein we examined the effects of yolk size and ovulation order on the fertilisation of yolks within double-yolked (DY) duck eggs. Single-yolked (SY) duck eggs had high fertility (89.98%), whereas in DY duck eggs fertility was low (51.9% yolks). The yolks closer to the airspace (Yolk 1) in DY eggs had higher fertility (68.82% vs 34.98% for Yolk 2; P<0.001). When only one of the two yolks was fertilised, Yolk 1 was fertilised in 99.12% of those eggs. Yolk 1 is presumed to be the first ovulated. The higher fertility of Yolk 1 is consistent with a primacy effect where the first ovulation has a higher probability of achieving fertilisation. Small DY eggs within the normal SY egg weight range (75-104.9g) had significantly lower yolk fertility (47.91%; n=550) compared with large (≥105g) DY eggs (54.56%; n=768; P<0.001). Yolk 1 fertility was lower in small compared with large DY eggs (64.18% vs 72.14%, respectively; P=0.003). Although Yolk 2 fertility was low, it was not affected by egg size. DY eggs with zero fertilised yolks were significantly lighter than DY eggs with one (P=0.007) or two (P<0.001) fertilised yolks (i.e. larger DY eggs were more likely to have fertilised yolks). Larger eggs (≥105g) have larger yolks and the evidence here shows that an optimal yolk size is a significant positive factor in achieving fertilization by dislodging spermatozoa, after ovulation, from their storage sites.