The reproductive physiology of the polyoestrous Honey possum (Tarsipes rostratus) is virtually unknown except that it shares with the kangaroos and wallabies the phenomenon of embryonic diapause. Its tiny size necessitates an alternate approach to study their reproductive cycle. We have accordingly utilised faecal steroid analysis. Baseline faecal cortisol levels in the Honey possum, at 4.1 +/- 0.3 mu g g(-1), are approximately 100-fold those of other mammals and are associated with adrenal glands that, on a mass-specific basis, are almost 10 times larger than the adrenals of other mammalian, including marsupial, species. Histological examination of the adrenal glands revealed no abnormalities, however, but their hypertrophy and the peaks recorded in faecal levels following disturbance suggest that the Honey possum is vulnerable to chronic stressors in the captive situation. Mean faecal progestagens (124.4 +/- 107.3 ng g(-1)) and oestradiol-17 beta (4.1 +/- 1.1 ng g(-1)) in 4 non-pregnant females maintained long term were not different from those of 5 pregnant females (101.4 +/- 61.0 ng g(-1) and 4.3 +/- 1.5 ng g(-1), respectively) and, on analysis, revealed a cyclicity of 24 +/- 1.2 days. We would predict from this evidence that the gestation period, in the absence of lactation, is approximately 23 days. Four of the pregnant females, monitored from July to November under conditions of 10:14 L:D photoperiod, showed a fall in levels of progestagens from 175.9 +/- 10.8 ng g(-1) in July and August to 30.9 +/- 9.4 ng g(-1) in October, while mean faecal levels of oestradiol-17 beta increased from 3.8 +/- 0.4 ng g(-1) in July to 5.7 +/- 0.3 ng g(-1) in October. September and October are months of peak reproductive activity in the wild and we suggest that these hormonal modulations may represent an entrained reproductive rhythm. Blastocysts appear to develop at varying rates, both within the one uterus, and between the two uteri of a single female. In addition, the time taken to reach the blastocyst stage may be longer than in any other marsupial studied to date. An association of the age of the pouch young with the stage reached by the developing blastocyst does not support the conclusion that blastocysts, once formed, grow slowly during lactation or diapause. Contrary to previous reports, we have documented what appears to be a lactational inhibition on blastocysts in diapause and have estimated the length of the 'delayed' reproductive cycle in two females as less than 2 weeks. Reactivation of blastocysts in Tarsipes has been shown to be stimulated by shortening day lengths after the summer solstice, a response similar to the annual breeding period of macropodid marsupials. Results from studying Honey possums in captive conditions suggest that the control of diapause in Tarsipes appears to be three-fold; lactational, photoperiodic and an entrained rhythm. (c) 2006 Elsevier Inc. All rights reserved.