Thesis (Ph.D.)--Murdoch University, 2004. / Thesis submitted to the Division of Science and Engineering. Bibliography: leaves 239-264.

Thesis (Ph. D) --Murdoch University, 2003. / Thesis submitted to the Division of Science and Engineering. Includes bibliographical references (leaves 275-282).

The common brushtail possum (Trichosurus vulpecula) is the most significant vertebrate pest in New Zealand as an ecological threat to the indigenous biodiversity and an economic threat as a vector for bovine tuberculosis. Biological control is considered to be the most accepted management strategy to reduce the population, specifically by impairing fertility. Successful development of a biocontrol agent (most likely a protein or peptide macromolecule) requires identification of a compound that is species-specific and potent. The challenge is also to deliver the bioactive to this free-ranging, widespread, feral animal and ensure sufficient bioavailability. Macromolecules have low oral bioavailability, thus new formulation strategies are required to enhance stability and absorption in the gastrointestinal tract (GIT) of T. vulpecula. Oral administration of the bioactive contained within a non-toxic bait is the most practical delivery strategy. Essential to designing an oral delivery system is to quantify the transit time of different sized delivery systems. The gastrointestinal transit in T. vulpecula was investigated (n = 72) by gamma scintigraphy. Technetium-labelled (99mTc) anion exchange resin particles (75 - 125 (mu)m or 500 - 700 (mu)m) or solution (99mTc-DTPA) was administered orally. After 3, 6, 12, 24 or 32 h, distribution of radioactivity in excised GITs was determined. Transit profiles were similar for each formulation. For delivery to the hindgut, bioactives need protection for 12 h though the upper GIT. Particulate formulations may be retained in the caecum for up to 32 h. Transit time was not different between animals dosed in the evening or the morning. Furthermore, GIT morphology is different between specimens in this study from southern New Zealand and Australian specimens. This may reflect improved diet quality in New Zealand. A model protein (insulin) was incorporated into poly(ethyl 2-cyanoacrylate) (PECA) nanoparticles prepared by interfacial polymerisation of water-in-oil microemulsions. The mean size of nanoparticles was 220 nm with a mean entrapment efficiency of 78%, determined using reverse phase HPLC. In vitro release of insulin from PECA nanoparticles in phosphate buffer (0.067 M, pH 7.4) at 37°C was triphasic and not all entrapped insulin was released. Following in vitro incubation of nanoparticles with enzyme solutions prepared from the GIT of T. vulpecula, lumen enzymes were more aggressive towards insulin compared to mucosal enzymes and the hindgut lumen was the GIT region with the lowest degradation. For the first time in a marsupial species, the in vivo pharmacokinetics of insulin-loaded, PECA nanoparticles were investigated following i.v. and intra-caecal administration and measured by radioimmunoassay. The low cross reactivity of human and endogenous brushtail possum insulin means that T. vulpecula is a suitable non-diabetic model to study pharmacokinetics of insulin. The i.v. pharmacokinetics of insulin solution and insulin-loaded nanoparticles were similar. On intracaecal dosing, co-administration of a permeation enhancer (EDTA) resulted in a small increase in plasma insulin concentration compared to insulin-loaded nanoparticles alone. In conclusion, transit time to the caecum of T. vulpecula following oral delivery was 12 h for fluid and particulate formulations < 1 mm diameter and was independent of the time of day the dose was given. T. vulpecula is a potential non-diabetic model for the study of insulin pharmacokinetics. This thesis demonstrates the potential application of oral peptide and protein delivery technology in the area of wildlife management.

The marsupial family Pseudocheiridae is currently known from seventeen species of six genera in Australia and New Guinea. These small to medium-sized arboreal animals are nocturnal and folivorous. Extinct pseudocheirids are recognised from several mid to late Cenozoic fossil localities across Australia and New Guinea. The single largest collection of pseudocheirid fossils has been recovered from the Oligo-Miocene freshwater carbonates of the Riversleigh World Heritage Area in northwest Queensland. This collection, which includes the first pseudocheirid cranial fossils, forms the basis of this investigation. Three new extinct pseudocheirid genera together containing four species are identified and described. Six new species of Paljara, Marlu and Pildra are also described from Riversleigh. Two of the new Marlu species are reported from South Australia??s Leaf Locality. From Riversleigh, Marlu kutjamarpensis is identified and additional material of Paljara tirarensae and P. nancyhawardae documented. New species attributed to Marlu and Pildra necessitate revision of those genera. Cranial material is identified for three of the new species. The rostrum of archaic pseudocheirids is shorter than in extant forms but cranial morphology is similar overall. Phylogenetic relationships of all extinct pseudocheirids are analysed. They include all new and previously described species, most of which have never been examined in a parsimony-based analysis. Two hypotheses of pseudocheirid evolution are presented: a paired lineage hypothesis and a single lineage hypothesis. Both hypotheses demonstrate that species of Paljara are not the most plesiomorphic pseudocheirids, Marlu praecursor does not cluster with other species of Marlu, the new genus Gawinga is most closely related to Paljara and there are no representatives of the extant genus Pseudochirops in any pre-Pliocene locality. All extant pseudocheirids cluster to form a crown clade sister to a stem lineage of Pseudokoala and Marlu species. Pseudocheirids are found in all Oligo-Miocene faunal zones of Riversleigh. Species of Paljara and Marlu are most frequently recovered from Faunal Zone B and C deposits respectively. Four pseudocheirid species biostratigraphically correlate the Kutjamarpu local fauna of the Leaf Locality with Faunal Zones B and C of Riversleigh, suggesting an early to middle Miocene age for both deposits. Modern pseudocheirids first evolved no later than the late Miocene from a descendant of the Marlu + Pseudokoala lineage when all other Oligo-Miocene pseudocheirids became extinct. At least three pseudocheirid lineages dispersed to New Guinea approximately five million years ago, but ecological barriers probably prevented subsequent migrations between the two landmasses.

The marsupial family Pseudocheiridae is currently known from seventeen species of six genera in Australia and New Guinea. These small to medium-sized arboreal animals are nocturnal and folivorous. Extinct pseudocheirids are recognised from several mid to late Cenozoic fossil localities across Australia and New Guinea. The single largest collection of pseudocheirid fossils has been recovered from the Oligo-Miocene freshwater carbonates of the Riversleigh World Heritage Area in northwest Queensland. This collection, which includes the first pseudocheirid cranial fossils, forms the basis of this investigation. Three new extinct pseudocheirid genera together containing four species are identified and described. Six new species of Paljara, Marlu and Pildra are also described from Riversleigh. Two of the new Marlu species are reported from South Australia??s Leaf Locality. From Riversleigh, Marlu kutjamarpensis is identified and additional material of Paljara tirarensae and P. nancyhawardae documented. New species attributed to Marlu and Pildra necessitate revision of those genera. Cranial material is identified for three of the new species. The rostrum of archaic pseudocheirids is shorter than in extant forms but cranial morphology is similar overall. Phylogenetic relationships of all extinct pseudocheirids are analysed. They include all new and previously described species, most of which have never been examined in a parsimony-based analysis. Two hypotheses of pseudocheirid evolution are presented: a paired lineage hypothesis and a single lineage hypothesis. Both hypotheses demonstrate that species of Paljara are not the most plesiomorphic pseudocheirids, Marlu praecursor does not cluster with other species of Marlu, the new genus Gawinga is most closely related to Paljara and there are no representatives of the extant genus Pseudochirops in any pre-Pliocene locality. All extant pseudocheirids cluster to form a crown clade sister to a stem lineage of Pseudokoala and Marlu species. Pseudocheirids are found in all Oligo-Miocene faunal zones of Riversleigh. Species of Paljara and Marlu are most frequently recovered from Faunal Zone B and C deposits respectively. Four pseudocheirid species biostratigraphically correlate the Kutjamarpu local fauna of the Leaf Locality with Faunal Zones B and C of Riversleigh, suggesting an early to middle Miocene age for both deposits. Modern pseudocheirids first evolved no later than the late Miocene from a descendant of the Marlu + Pseudokoala lineage when all other Oligo-Miocene pseudocheirids became extinct. At least three pseudocheirid lineages dispersed to New Guinea approximately five million years ago, but ecological barriers probably prevented subsequent migrations between the two landmasses.

The honey possum Tarsipes rostratus is a tiny (7 - 12 g) highly specialised flower-feeding marsupial endemic to the south-western corner of Australia. The impact of fire on this small mammal was studied, over a 19-year period, in the Fitzgerald River National Park, a large (330,000 ha) area of relatively undisturbed heathland/shrubland, rich in the proteaceous and myrtaceous plants upon which the honey possum appears to rely for food. The honey possum is the most abundant and widespread mammal in this Park. Capture rates of honey possums were significantly related to the years since the vegetation was last burnt, annual rainfall in the preceding (but not the current) year, the season when trapping occurred, and the trapping grid operated. Capture rates declined markedly after fire and remained low (less than one third of those in long unburnt vegetation) for about 4 - 5 years following a fire. Rates of capture then increased steadily over the next 20 - 25 years, with maximal abundance recorded about 30 years after fire. Thereafter, there appeared to be a slight decline in capture rates, but even in the vegetation unburnt for longest (> 50 years since fire), honey possum abundance was substantial and relatively stable. In contrast to these changes in abundance, the structure of the honey possum population, with 79 % adults and 57 % males, appeared little influenced by fire history, annual rainfall, season or grid. The increase in the rates of capture of honey possums following fire paralleled the pattern of availability of cover in the vertical and, to a lesser extent, horizontal plane. Indeed, projective foliage cover took around 20 years after fire to reach levels similar to those available in areas unburnt for even longer. The trend in capture rates was also congruent with the maturation of the most frequently visited foodplants of honey possums, particularly Banksia nutans (summer flowering) and B. baueri (winter flowering). Areas long unburnt still contained shelter and foodplants adequate for honey possums even 50 years or more after fire, with only slight evidence of senescence. Pollen loads indicated that honey possums caught in burnt areas, where their preferred foodplants were absent, continued to feed on these favoured foodplants (Banksia and Dryandra spp.) at nearby unburnt areas. In addition, they also fed, in both burnt and long unburnt areas, upon a suite of other plant species that regenerated more rapidly from lignotubers and epicormic buds, as well as from seeds (e.g. Eucalyptus and Calothamnus spp.). Thus, honey possums appeared to persist with their preferences for feeding from a limited number of flowering plants despite some of these species not being available in recently burnt areas for many years. Nearby patches of unburnt vegetation can clearly be important refuges, feeding grounds and shelter for the few honey possums that visit recently burnt areas, and appear to be the source of honey possum colonists in the years following a fire. Capture rates were also greater following years when rainfall was higher than average. Indeed, rainfall had as great an influence upon capture rates as time since fire. Capture rates were also consistently higher over winter, and to a lesser extent over summer, than in either autumn or spring. Individual grids, even those close together in apparently similar vegetation with a similar fire history, still differed significantly overall in their capture rates of honey possums. This last finding has implications for the use of chronosequences in the study of post-fire changes in biota. Although not the primary focus of the study, data on the limited suite of other, far less abundant, small mammals present indicated that house mouse Mus musculus domesticus numbers peak soon after fire (about two years after fire), grey-bellied dunnart Sminthopsis griseoventer numbers somewhat later (about eight years after fire) and that southern bush rats Rattus fuscipes fiuscipes, like honey possums, are later successional species. Most species were present in vegetation over a range of post-fire ages, with data consistent with models based on sequential changes in relative abundance. Like many Australian mammals, the range of the honey possum has contracted substantially over the last 200 years and the coastal heathlands of the south-west are its last stronghold. In terms of its conservation, this study indicates that, if possible, management burns in these heathlands should be separated by intervals of at least 20 years between successive burns, and preferably even longer. If burns are required more frequently to meet other management priorities, it is highly preferable that they are small and patchy, rather than large scale. Such practices may help ensure the long-term survival of this unique, highly specialised and endemic marsupial.

A vegetation assessment was conducted to understand the effect of feral goats (Capra hircus) on the vegetation of Isolated Hill Scenic Reserve located in Southern Marlborough. New Zealand forests evolved without the pressure of mammalian herbivores but following human settlement they became subject to intensive browsing following the introduction of exotic mammals. This study focuses on the presence of feral goats. Interest arises from the settlement of other countries and the subsequent liberations of feral goats which are now considered to be responsible for the significant removal of native vegetation as well as playing a dominant role in erosion. Changes in plant communities have occurred with the pressure of goat browsing as well as secondary effects such as habitat degradation The purpose of this research was to investigate the changes in plant species composition since the initial and subsequent research conducted in 1985 and 1994. An analysis of previous work on the diet of multiple introduced ungulate species was also conducted. Previous studies were not limited to only feral goat studies and included other introduced herbivores to provide a broad overview of diets. The New Zealand Forest Service implemented a programme for monitoring the seral forest in Isolated Hill Scenic Reserve using standard Forest Service vegetation quadrants. 32 quadrants were established, including three exclosures, in various locations around the Reserve to allow assessment of feral goat impacts on seral forests (forest whose constituents are of varied age classes). Nine forest service plots, including three exclosures were re-measured and an additional nine plots were measured. Given the progression of vegetation change that has been observed during the course of this study, both analytically and observationally, Isolated Hill Scenic Reserve in its current state, involving the notable presence of feral goats and the vegetation types will continue to diminish in diversity.

This study investigates the ecology of one of the best known Australian marsupials, the Common Brushtail Possum Trichosurus vulpecula, in central Australia. Trichosurus vulpecula is one of few medium-sized mammal species that persist in arid Australia today. Its distribution within the arid zone has declined markedly since European settlement. Two populations, one within the East MacDonnell Ranges along the Hale River and the other on Irving Creek, a River Red Gum creek in the Petermann Ranges, were studied in the southern Northern Territory. Others locations in the region were visited opportunistically. Trie central Australian Trichosurus is not distinct genetically from populations elsewhere in Australia. The diet of T. vulpecula consisted of a range of leaves, flowers and fruits of perennial dicotyledonous species as well as some ephemeral herbs. Grasses were absent from the diet. Variation in the diet reflected seasonal availability in flowers and fruits. The species preferentially consumed at each site had significantly higher moisture content and dry matter digestibility than species not consumed. Preferred species included Amyema maidenii leaves (a mistletoe), Acacia spp. flowers and fruits, Santalum lanceolatum leaves (a shrub), Marsdenia australis leaves (a vine), Solarium quadriloculatum fruit (shrub) and Euphorbia spp. leaves (herb). Small amounts of invertebrate material were consumed throughout the year. Other non-plant material consumed included honeycomb and unfledged birds eg. Budgerigars. There were no significant differences in the diet between the sexes. Trichosurus vulpecula were found in six main habitats: Acacia aneura/Callitris glaucophylla on rocky hills; E. camaldulensis sandy creek-lines; mixed Acacia rocky hills, Rocky Eucalyptus creek-lines; Degraded drainage lines; and Wet gullies. Logistic regression modelling revealed a significant correlation between mistletoe species richness, higher levels of soil nitrogen and the presence of T. vulpecula. In habitats occupied by T. vulpecula species richness of mistletoes was associated with the absence of fire and the presence of reliable ground water supplies. Trichosurus vulpecula were highly mobile with mean home ranges at Hale River of 44.21 � 22.76 ha and considerably higher than those recorded in previous studies in Australia. Mean home ranges at Irving Creek were much smaller, at 4.99 � 1.46 ha and VII similar to that recorded in other studies in Australia. At both sites, males had larger home ranges and there was a high degree of overlap with other males and females. At the Hale River study site, T. vulpecula predominantly denned in caves or cavities in rocks, whereas at Irving Creek all den sites were in large Eucalyptus camaldulensis on the drainage line. Adult and pouch young sex ratios were at parity. During this study, T. vulpecula was found to breed continuously, with births recorded in almost all months. Growth of the young were more rapid than previously recorded for Trichosurus in Australia. This is interpreted as an adaptation for living in an arid environment, enabling the young to achieve independence before quality food supplies diminish. No single exotic predator or competitor was solely responsible for the decline of T. vulpecula in arid Australia, implying an interactive impact. Prey switching by dingoes from rabbits to T. vulpecula, macropods and echidnas followed the crash of rabbit populations at Hale River. Predation by dingoes on T. vulpecula was only recorded once, at the Irving Creek study site, where numbers of rabbits remained stable throughout the study. The impact of exotic herbivores occurred through habitat degradation rather than competition. Evaluation of the ecological data collected during this study generally supports current models of decline and extinction in medium-sized mammals in arid Australia, integrating the effects of predators, competitors, drought and fire. However, the importance of each factor on populations of T. vulpecula was found to vary depending on their location in the landscape. This study suggests two separate models to explain the decline of T. vulpecula in arid Australia after the arrival of Europeans. The first operates in the riparian lowlands and the second on the rocky ranges. In both models, prior to European settlement, T. vulpecula occupied refuge habitats characterised by readily available moisture for plant growth (run on areas and/or shallow water tables) and soils with higher soil nutrient concentrations. The impact of fires on these refugia was minimal, as Aboriginal burning practices protected them with mosaic burning generally preventing large-scale fires from developing. Following European settlement, the forces impacting on populations were different in the riparian lowlands from those affecting rocky ranges. In the riparian lowlands, the effects of rabbits and livestock together with predation were found to have the major impact on T. vulpecula populations. Fire was not a significant factor in these areas. In the rocky ranges, fire was the most significant factor affecting T. vulpecula populations. Introduced herbivores did not degrade these habitats as they did in the riparian lowlands because the rugged and steep nature of the ranges acted as a physical barrier. Similarly, predator numbers were lower because of the relative difficulty in moving over rough ground and the generally lower relative abundance of preferred prey such as rabbits. An adaptive management strategy needs to be implemented to determine the effects of different management regimes on T. vulpecula population viability. The key elements of a management strategy in the riparian lowlands involves the manipulation and monitoring of predators, rabbits and livestock numbers. In the rocky ranges, the key management strategy involves the implementation of a patch burning to prevent fires entering habitats occupied by T. vulpecula. Importantly, any management strategies should involve Aboriginal people. Trichosurus vulpecula is an important part of Aboriginal culture. Its decline is of great concern to many people and several of the remaining populations and potential reintroduction locations are on Aboriginal land. Because of their relationship with the land and the animals, people have both the knowledge of the animal and the skills (such as patch burning) to provide information to managers which will assist with management. To achieve these management directions a coordinated national education programme is required to inform and convince the Australian community that conservation of T. vulpecula is deserving of attention in arid and semi-arid Australia. This is particularly important given the perception that T. vulpecula is a common species throughout Australia, despite its massive decline in arid Australia since European settlement.

Parasitism, in nematodes, is a very successful life strategy which has evolved throughout the Nematoda phylum in several independent events. However, the genetic basis for parasitism remains unknown. Parastrongyloides trichosuri is a facultative parasitic nematode of the Australian brushtail possum. This parasite has retained the unusual ability to sample its environment at each generation, and make the developmental decision to develop either into a free-living nematode or, in response to environmental stress, develop into an infective larva, which must then seek out a host in order to complete its life cycle. The nematode model organism, Caenorhabditis elegans, also responds to environmental stresses by developing into a dauer larvae. The dauer hypothesis proposes that dauer larvae and infective larvae are homologous and that dauer larvae may be an evolutionary pre-adaptation that facilitated the evolution of parasitism in nematodes. One of the signalling pathways which control dauer larva development in C. elegans is the Insulin/IGF signalling pathway. Gene orthologues of the insulin/IGF signalling pathway were cloned from P. trichosuri: the daf-2 tyrosine kinase receptor, the age-1 phosphatidylinositol 3′ kinase and the daf-16 FOXO forkhead transcription factor. The expression profiles of these genes were characterized by q-PCR which determined that they were differentially expressed during the developmental switch to infective larva. Rescue by complementation showed that a P. trichosuri daf-16 transgene was able to recover both stress and developmental phenotypes in C. elegans daf mutants, suggesting it might perform an orthologous role in P. trichosuri. This research also demonstrated that the biology of P. trichosuri infective larvae and C. elegans dauer larvae are quite similar. Some of the environmental signals which control the free-living/infective larva developmental switch in P. trichosuri were characterized in this study and found to be similar to the environmental signals which trigger dauer larval development. These are: population density, food availability and temperature. There is a genetic component to the ability to respond to the environmental signals and inbred lines which display diverse developmental plasticity were isolated.

The honey possum Tarsipes rostratus, a marsupial endemic to South-Western Australia, feeds exclusively upon nectar and pollen. It is one of the smallest marsupials, with adult females (8-12g) significantly larger than adult males (6-9g). Honey possum males have the longest sperm (356µm) recorded for any mammaland the testes represent 4.2% body weight, amongst the largest recorded formammal species. These features suggest that sperm competition is an importantpart of the mating system. This study used a combination of field based studies,DNA analysis and histological examination of the female reproductive tract toinvestigate the life history, multiple paternity and reproduction of the honey possum innatural populations in the Fitzgerald River National Park (FRNP), on the south coastof Western Australia. This study drew upon earlier work on the honey possum in the FRNP in order to describe its life-history. The honey possum is short-lived (1-2 years), and attains sexual maturity whilst still growing. All four teats are occupied after birth, but the litter is reduced to 2 or 3 young during pouch life. The young have a relatively slow rate of growth. Breeding occurs continuously throughout the year, but is affected by the flowering phenologies of its foodplants. The greatest proportion of females with pouch-young occurs in winter; there are fewest pouch-young in autumn, a time of year when there is a dearth of flowers. Honey possums are essentially solitary animals, with no structured social unit, and male and female home ranges overlap. In captivity they are largely tolerant of one another, but larger females are behaviorally dominant to smaller females and to males. The densities and structure of the honey possum populations in the FRNP were analyzed from trapping data collected over 19 years. Population densities fluctuated significantly from season to season throughout the year, with changes in the flowering food resources available. There were also year-to-year differences in the intensity of those fluctuations, and these were significantly associated with rainfall in the previous year, and probably mediated through a lag effect in the flowering of the honey possum’s foodplants. The greatest densities of animals occurred over winter. In years following high rainfall, mean winter densities reached 88 individuals per hectare. The lowest densities occurred in spring, and in years following low rainfall mean spring densities fell to 8 individuals per hectare. Even at these lowest densities, there is still the potential for interaction between males and females. A succession from high to low, then back to high densities was seen during the three years of the present study (2000-2002) and this shadowed a similar succession of changes in rainfall. The proportion of females with pouch-young was significantly affected by the season, and by rainfall in the previous year. Years following low rainfall had a lower proportion of females in a condition to breed. The autumn dip in breeding that occurred in all years was exacerbated following dry years. Of those females that did breed in 2001, a time of low resources, there was no difference in the size of the litter compared to 2000 and 2002, times of higher resource availability. The sex-ratio of pouch young was at parity, but there was a slight bias towards males among both juveniles (56%) and adults (58%). This was probably due to the greater movements shown by males. Sex ratios were not affected by changes in rainfall and density. Male-biased dispersal was detected using genetic data and the movement patterns of males showed that they moved greater distances than females during their normal activity. Analysis of four microsatellite loci revealed extremely high levels of variation, with 28 to 50 alleles per locus and a mean expected heterozygosity of 0.95. These are amongst the highest seen in any microsatellite study of vertebrates. There was multiple paternity in 86% litters, using a minimum number of sires per litter method, and in 95% litters, using an estimated number of sires method based upon the relatedness of litter males. This indicates that multiple mating is frequent in female honey possums and is evidence for sperm competition. The estimated number of sires in a litter was often three or four. In 41% of cases, the number of sires was less than the number of young in the litter, indicating that some males were more successful at siring offspring than others. Nevertheless, no more than two offspring in a litter were known to have been sired by the same male. Despite marked fluctuations in density from high in 2000, to low in 2001, then high again in 2002, the level of multiple paternity remained equally high in all years. Embryonic diapause and female reproduction was investigated in the honey possum. All adult females examined, both with and without pouch-young, were either close to oestrus, had ovulated or were carrying conceptuses. The honey possum has a postpartum oestrus and it was evident that this occurs approximately 2-4 days after birth. Cleavage and formation of the unilaminar blastocyst appears to occur rapidly over approximately 5 days. Embryonic diapause proceeded in a two phase manner similar to other small possum species. The unilaminar blastocyst expanded rapidly at first; and then, from about 18-20 days after birth, the diameter of the blastocyst remained constant at approximately 1.2-1.8mm. No growth or development beyond the unilaminar stage was observed during pouch-life. The first signs of reactivation occurred during lactation, after pouch exit, and expansion of the blastocyst only occurred in one post-lactational female. The development of the corpus luteum appeared different to patterns described for other marsupials, but its formation coincided with the formation of the unilaminar blastocyst. The diameter of the corpus luteum remained constant throughout diapause. The histology of the reproductive tract was generally similar to other marsupials. There were no sperm storage crypts in the female reproductive tract. The length of pouch-life in the honey possum was 55-65 days, and the interval between litters of the same size varied between 65 and 100 days. Embryonic diapause may reduce the time between production of successive litters in the honey possum, but lifetime reproductive potential is reasonably low. Females had up to four litters over the period that they were captured. Thus, each litter represents a substantial proportion (25%) of a female’s lifetime reproductive output. Reproductive amortization occurred, with 61% loss overall, due to overproduction of ova, loss of conceptuses and reduction of the litter during lactation. The behavioural dominance of females suggests that multiple mating is an active strategy, and this presumably allows the genetic quality of their offspring to be maximized. Males that succeed in sperm competition may be of better intrinsic quality. Overproduction of conceptuses by females presents the opportunity for them to select those fertilized by intrinsically viable males or genetically compatible males. Sexually active males are present all year round. Females were not synchronous in their sexual receptivity, and this would lead to a skewed operational sex ratio, with more reproductive males than oestrous females. Since adult males are significantly smaller than adult females and possess no ornaments or armaments, it is unlikely that males overtly fight for access to females. Rather, males appear to monitor the reproductive status of females through smell, and probably compete in their ability to locate oestrous females. The risk and intensity of sperm competition is high, sexual selection for a large investment in spermatogenesis is evident and competition after copulation is probably an important factor in the mating system. It is likely that males, as well as females mate multiply, and the mating system is promiscuous.