Antechinus agilis mate within a 2–3 week, highly synchronized period each year, with copulation consisting of short bouts of thrusting interrupted by longer bouts of rest during which time the male remains mounted. In the laboratory, mating can last as long as 8–12 h without any break in intromission, with captive paired animals generally copulating once per day (Woolley, 1966a,b). The mating programme used in this study examines the effects on copulatory behaviour of changing: (1) timing of access relative to ovulation; (2) order of mating; (3) the delay between the first and second males' mating access. The total mount time was divided into quarters and the changes in behavioural patterns assessed by examining changes in the frequency of five key activities; thrusting, pelvic side-to-side movements, walking, female resistance, and dismounts. No significant differences were observed in the time from initial pairing to first mount regardless of mating order or time of mating, nor was the total time mounted significantly affected by mating order. Significant reductions in the total time mounted were evident, however, for those males mating closer to the time of ovulation. The behavioural strategies associated with copulation in A. agilis significantly enhance arguments for equality between males and females in determining overall mating strategies.

The mouse-like marsupial Antechinus agilis is common to south-eastern Australia with breeding and life-history traits being highly synchronous. Mating activity is confined to a 10- to 15-day period in August, at the end of which all males die as a result of a stress-induced suppression of the immune system. Ovulation occurs at this time and females rely on stored sperm from specialized crypts in the lower isthmus of the oviduct for successful fertilization. Here we report a high incidence of mixed paternity litters, which can be attributed to sperm from multiple males being stored in the isthmic crypts. Data from this study also suggest a possible second male siring advantage in controlled ex situ sperm competition mating trials, irrespective of the delay between the two males being given mating access to the female or of the mating time relative to ovulation. In determining paternity through DNA profiling, population genetic data were obtained that showed significant differences in the genetic heterozygosity between unrelated adults, half siblings and full siblings. We suggest that the isthmic crypts, in addition to storing viable sperm, are capable of releasing a mix of sperm that increases the likelihood of mixed paternity litters. This allows all mating males the opportunity to sire young, increases female reproductive fitness and overall maintains high levels of population genetic heterozygosity in the face of total male mortality annually.