Behavioural and genetic assessment of reproductive success in a spawning aggregation of the Australian giant cuttlefish, Sepia apama

The only known spawning aggregation of cuttlefish occurs in winter in southern Australia. The operational sex ratio in this aggregation is highly skewed towards males (range 11:1 to 4:1). Using SCUBA, we videotaped females as they mated with up to six males/h. Females rejected and accepted mates of any status or size. We obtained individual eggs and tissue samples from females and their mates and determined paternity by microsatellite DNA analysis. Thirty-nine eggs were sampled during 17 observational sequences (trials); 22 eggs were sired by a sampled male. One-third of the females mated with multiple males and 67% of females' eggs had multiple sires (N=9). Given our restricted sampling, these results probably underestimate actual levels of multiple paternity. Males competed to guard and copulate with females, and spent most of the copulation time flushing water into the female's buccal area. Thus, we predicted a high probability of fertilization of the next-laid egg(s) by the last male to mate, but this prediction was not statistically supported by our data. Fertilization success did not differ significantly between paired consorts (36%) and unpaired males (31%), or between large (33%) and small males (29%). Females used sperm from paired, unpaired and sneaker males as well as from previous matings to fertilize eggs. Our results indicate that this mating system has a high level of multiple mating and multiple paternity and that males of any size or status can obtain successful fertilizations.