Here we present the genetic relationships of 26 specimens of the genus Breinlia (Nematoda: Filarioidea) from a range of Australian marsupials using markers in the small subunit of nuclear ribosomal RNA and mitochondrial cytochrome c oxidase subunit 1 (cox1) genes and compare them with morphological determinations. The molecular data support the validity of most of the morpho-species included in the study and provide provisional insights into the phylogeny of the genus in Australian mammals, with dasyuroid marsupials appearing to be the original hosts. The recent discovery of Breinlia annulipapillata in the eye of a human brings this genus of parasites into the group of emerging infectious parasitic diseases.

An important component of reintroduction is acclimatization to the release site. Movement parameters and breeding are common metrics used to infer the end of the acclimatization period, but the time taken to locate preferred food items is another important measure. We studied the diet of a reintroduced population of brushtail possums Trichosurus vulpecula in semi-arid South Australia over a 12 month period, investigating changes over time as well as the general diet. We used next-generation DNA sequencing to determine the contents of 253 scat samples, after creating a local plant reference library. Vegetation surveys were conducted monthly to account for availability. Dietary diversity and richness decreased significantly with time since release after availability was accounted for. We used Jacob's Index to assess selectivity; just 13.4% of available plant genera were significantly preferred overall, relative to availability. The mean proportion of preferred plant genera contained within individual samples increased significantly with time since release, but the frequency of occurrence of preferred plants did not. Five genera (Eucalyptus, Petalostylis, Maireana, Zygophyllum and Callitris) were present in more than half of samples. There was no difference in dietary preferences between sexes (Pianka overlap = 0.73). Our results suggest that acclimatization periods may be longer than those estimated via reproduction, changes in mass and movement parameters, but that under suitable conditions a changeable diet should not negatively affect reintroduction outcomes. Reintroduction projects should aim to extend post-release monitoring beyond the dietary acclimatization period and, for dry climates, diet should be monitored through a drought period.

Trypanosomes are blood-borne parasites that can cause severe disease in both humans and animals, yet little is known of the pathogenicity and life-cycles of trypanosomes in native Australian mammals. Trypanosoma copemani is known to be infective to a variety of Australian marsupials and has recently been shown to be potentially zoonotic as it is resistant to normal human serum. In the present study, in vivo and in vitro examination of blood and cultures from Australian marsupials was conducted using light microscopy, immunofluorescence, scanning electron microscopy and fluorescence in situ hybridization. Promastigote, sphaeromastigote and amastigote life-cycle stages were detected in vivo and in vitro. Novel trypanosome-like stages were also detected both in vivo and in vitro representing an oval stage, an extremely thin stage, an adherent stage and a tiny round stage. The tiny round and adherent stages appeared to adhere to erythrocytes causing potential haematological damage with clinical effects similar to haemolytic anaemia. The present study shows for the first time that trypomastigotes are not the only life-cycle stages circulating within the blood stream of trypanosome infected Australian native marsupials and provides insights into possible pathogenic mechanisms of this potentially zoonotic trypanosome species.

A total of 41 ticks were collected from 15 quokkas on Bald Island and 2 ticks from a Gilbert's potoroo from Two Peoples Bay. Three species of Ixodid ticks Ixodes australiensis, Ixodes hirsti and Ixodes myrmecobii were identified on the quokkas known to have a high prevalence of Trypanosoma copemani. Tick faeces from ticks isolated from 8 individual quokkas and a Gilbert's potoroo were examined with one identified as positive for trypanosomes. Faecal examination revealed trypanosomes similar to in vitro life-cycle stages of T. copemani. In total 12 ticks were dissected and trypanosomes found in sections of their midgut and haemolymph, 49 and 117 days after collection. Tick faeces, salivary glands and midguts from I. australiensis were screened using an 18S rRNA PCR with amplification seen only from the midguts. Sequencing showed 100% homology to T. copemani (genotype A) and 99·9% homology to the wombat (AII) isolate of T. copemani. Trypanosomes were only detected in I. australiensis as neither I. hirsti nor I. myrmecobii survived the initial 30-day storage conditions. We therefore identify a vector for T. copemani as I. australiensis and, given the detection of trypanosomes in the faeces, suggest that transmission is via the faecal-oral route.

Differences in growth patterns between the sexes of the gracile mouse opossum Gracilinanus microtarsus and the consequences for home range size were investigated in a savanna habitat (cerrado) of south-eastern Brazil. A total of 51 juvenile individuals of Gracilinanus microtarsus was monitored using capture–mark–recapture from November 2005 to August 2006. The increase in body mass of gracile mouse opossums was described using the Gompertz growth model. Male gracile mouse opossums grew faster than females (dimorphic ratio of 1.5). Home range size, estimated with the minimum convex polygon method, was positively related to body mass. Model selection using Akaike's Information Criterion (AICc) and incorporating body mass, sex and season as independent variables showed that the best-supported model describing variance in home range sizes included only body mass. Our data suggest that a greater body mass gain in juvenile males is probably the proximate cause of sexual dimorphism in adult gracile mouse opossums and that energetic needs required for growth have a greater influence in home range size.

To date, little is known about the dynamics of vertical transmission of Toxoplasma gondii in Australian marsupials. Studies in mice demonstrate that vertical transmission of T. gondii is common and that chronically infected mice can transmit T. gondii to successive generations. In this study, PCR and immunohistochemistry were used to detect T. gondii in chronically infected marsupial dams and their offspring. T. gondii was detected in the unfurred pouch young of 2 out of 10 chronically infected western grey kangaroos (Macropus fuliginosus) and in the unfurred pouch young of a brush-tailed bettong (Bettongia penicillata). Results of the study suggest that vertical transmission of T. gondii can occur in chronically infected Australian marsupials.

The retinas of placental mammals appear to lack the large number and morphological diversity of cone subtypes found in diurnal reptiles. We have now studied the photoreceptor layer of a South American marsupial (Didelphis marsupialis aurita) by peanut agglutinin labeling of the cone sheath and by labeling of cone outer segments with monoclonal anti-visual pigment antibodies that have been proven to consistently label middle-to-long wavelength (COS-1) and short-wavelength (OS-2) cone subpopulations in placental mammals. Besides a dominant rod population (max. = 400,000/mm2) four subtypes of cones (max. = 3000/mm2) were identified. The outer segments of three cone subtypes were labeled by COS-1: a double cone with a principal cone containing a colorless oil droplet, a single cone with oil droplet, and another single cone. A second group of single cones lacking oil droplets was labeled by OS-2 antibody. The topography of these cone subtypes showed striking anisotropies. The COS-1 labeled single cones without oil droplets were found all over the retina and constituted the dominant population in the area centralis located in the temporal quadrant of the upper, tapetal hemisphere. The population of OS-2 labeled cones was also ubiquitous although slightly higher in the upper hemisphere (200/mm2). The COS-1 labeled cones bearing an oil droplet, including the principal member of double cones, were concentrated (800/mm2) in the inferior, non-tapetal half of the retina. The two spectral types of single cones resemble those of dichromatic photopic systems in most placental mammals. The additional set of COS-1 labeled cones is a distinct marsupial feature. The presence of oil droplets in this cone subpopulation, its absence in the area centralis, and the correlation with the non-tapetal inferior hemisphere suggest a functional specialization, possibly for mesopic conditions. Thus, sauropsid features have been retained but probably with a modified function.

Little is known of the prevalence and life-cycle of trypanosomes in mammals native to Australia. Native Australian trypanosomes have previously been identified in marsupials in the eastern states of Australia, with one recent report in brush-tailed bettongs (Bettongia penicillata), or woylie in Western Australia in 2008. This study reports a novel Trypanosoma sp. identified in blood smears, from 7 critically endangered Gilbert's potoroos (Potorous gilbertii) and 3 quokkas (Setonix brachyurus) in Western Australia. Trypanosomes were successfully cultured in vitro and showed morphological characteristics similar to members of the subgenus Herpetosoma. Phylogenetic analysis of 18S rRNA gene sequences identified 2 different novel genotypes A and B that are closely related to trypanosomes previously isolated from a common wombat (Vombatus ursinus) in Victoria, Australia. The new species is proposed to be named Trypanosoma copemani n. sp.

In Australia, macropodids are common intermediate hosts for the cestode Echinococcus granulosus, and sylvatic transmission is maintained via wild dogs. The parasite causes mortality in a number of macropodid species and the sylvatic cycle provides a source of infection to domestic livestock and humans. We determined the efficacy of the hydatid vaccine, EG95 in the tammar wallaby, Macropus eugenii, challenging either 1 or 9 months post-vaccination. EG95 provides similar protection to that seen in sheep (96–100%). Control tammars were significantly more likely to become infected (odds ratio 29·44; CI 4·13, 209·97; P=0·001) and to develop more cysts (count ratio 26·69; CI 5·83, 122·19; P<0·001). The vaccination may be beneficial if administered pre-release in captive breeding programmes for endangered macropodids. Further work to develop oral delivery methods may enable vaccine administration of wild animals and thereby a reduction in sylvatic transmission.

The morphology of the acrosome of spermatozoa an Australian daysyurid marsupial during in vivo fertilisation, as seen with the tranamission electron microscope, is described. In some spermatozoa close to the outer zona surface with largely intact acrosomes, eletron-demse struts between the inner and outer acrosomal membranes could be seen; in addition fusion between the outer acrosomal and overlying plasma membranes was sometimes evident. The zona matrix close to the acrosomal region of spermatozoa with heads party embedded within the zona pellucida was less electron-dense and more filamentous than elsewhere; perhaps indicative of local lytic activety. Nevertheless some of these spermatozoa at least had partly intact acrosomes with local regions of electron-dense bridges between the two acrosomal membranes. Spermatozoa with condensed chomatin were also observed within the egg cytoplsam some of which had membranes, vacuoles or close to the dorsal nuclear surface which could, in part, be acrosomal in orgin. These observations lend some support to the recent view, based on findings from an American species, that acrosomal changes of spermatozoa of some marsupials at the of fertillsation may be somewhat similar to those of eutherian mamals in spite of the considerable differences in gamete structure and design. How generally applicable these findings are to all marsupials remains, however, to be determined.

The occurrence of in vivo parthenogenesis is documented for laboratory-bred individuals of the Australian dasyurid marsupial, Sminthopsis crassicaudata. About 30% of females that had been isolated from males for a greater period of time than the length of pregnancy were found, on dissection of their uteri, to have embryos present. The embryos were surrounded by a mucoid coat and shell membrane and at least the first two cleavage divisions occurred normally. After this time, however, unequal cleavage divisions appeared to result. The parthenogenesis that takes place may be initiated as a result of activation of ageing ovulated oocytes. Its frequent occurrence may prove useful in a study of maternal and paternal contributions to early embryonic development in this species.

Even though the great majority of the biomass and diversity of mammals in tropical forests inhabit the canopy (Eisenberg & Thorington 1973), most knowledge of this group is based on forest-floor samples (Lowman & Moffett 1993). Studies that include trapping efforts in the canopy are becoming increasingly common (Grelle 2003, Lambert et al. 2005, Malcolm 1995, Patton et al. 2000, Vieira & Monteiro-Filho 2003, Voss et al. 2001), but aspects on the ecology of arboreal small mammals still remain poorly understood. Many species of non-flying mammals co-occur in the canopy and, thus, are expected to use niche dimensions differently to permit coexistence (Cameron et al. 1979). Despite the difficulties of access to and in understanding the three-dimensional use of the arboreal strata by the different species (Emmons 1995), some studies have demonstrated that the differential use of the habitat (Cunha & Vieira 2002, Gentile & Fernandez 1999), of food items (Cáceres et al. 2002, Santori et al. 1995) or both (Leite et al. 1996, Mauffrey & Catzeflis 2003), include strategies involved in the division of resources.

Infection of small macropodids with the larval stage of Echinococcus granulosus can cause fatalities as well as significant pulmonary impairment and other adverse sequelae. The brush-tailed rock-wallaby (Petrogale penicillata) is a small macropodid listed as vulnerable on the IUCN's Red List of Threatened Species. This study used radiographic techniques to determine the prevalence and severity of pulmonary hydatid infection and growth rates of hydatid cysts in a wild population of this macropodid. The overall prevalence was 15·3% (9/59 animals) with 20·0% (8/40 animals) of adults infected. During the study period, the death of at least 1 infected animal was directly attributed to pulmonary hydatidosis. Rapid cyst growth occurred in some animals (up to 43% increase in cyst volume in 3 months). Cyst volume reduced lung capacity by up to 17%. Secondary pulmonary changes were uncommon but, in 1 animal, resulted in reduction in lung capacity by approximately 50%. Infection was associated with a higher blood urea concentration, but no significant differences in other blood variables were detected. These results indicate that hydatid infection may be a significant risk to threatened populations of small macropodids and should be addressed in conservation management plans for these animals.

The temporal pattern of reproduction and its consequences for age structure and density were investigated in a population of the gracile mouse opossum Gracilinanus microtarsus in south-eastern Brazil. Individuals of G. microtarsus were monitored through capture–mark–recapture methods from August 2000 to February 2003 in a remnant of cerradão, a forest-like physiognomy of the highly seasonal cerrado biome. The temporal pattern of reproduction of the population studied was highly seasonal with rearing of the offspring occurring in the first half of the warm-wet season, when the abundance of food resources – primarily insects – in the cerrado is high. Shortly after reproduction, the density of adults decreased sharply, possibly because of high post-mating mortality, leading to a gradual replacement of adults by their offspring in the following months and little overlap of generations. Our data suggest that climatic and environmental factors affect the onset of reproduction and interact with endogenous factors that decrease post-mating survival to produce the observed pattern of seasonal variation in age structure and density. It is suggested that the dynamics of populations of G. microtarsus may be driven primarily by food limitation and that long-term studies are needed to understand its feedback structure.

Little is known of patterns of male reproductive effort and investment, and their relationship with age-related differences in male mating strategy, in mammals. In this study, the age-specific patterns of reproductive expenditure in a male marsupial, the common brushtail possum Trichosurus vulpecula were investigated. Possums were studied for two breeding seasons on Magnetic Island, north Queensland, Australia. Energy expenditure in male possums during the mating season was estimated from changes in mass taken monthly during live-trapping. Male possums lost an average of 4.2% of their mass during the main mating season from January to May, suggesting that mating and consort behaviour is energetically expensive and may constrain foraging activity. The percentage of body mass lost by individual males during the breeding season was negatively related to age; older males directed less somatic investment into reproduction than young males. Older males seemed to target their energy expenditure toward the period when oestrus synchrony was highest. The pattern of age-specific energy expenditure for reproduction in T. vulpecula differs markedly from that reported previously for male cervids, probably owing to the spatial dispersal of females leading to differences in mating strategy. The results are discussed in terms of their relevance to potential age-related differences in reproductive strategy and behaviour.

Most mammals exhibit seasonal variation in the reproductive capacity of one or both sexes. While the female southern hairy-nosed wombat Lasiorhinus latifrons is a known seasonal breeder, the extent of seasonality in the male has not been documented. To examine this, gross body measurements including scrotal diameter and the dimensions of the accessory gland bulge were recorded and male reproductive tracts were examined between 1993 and 2000. Testes, epididymides and accessory glands from all males were dissected free of connective tissue and weighed. In addition, matched semen samples were collected over four time points in 2000 corresponding to the breeding season (September), immediately post-breeding season (November), during the non-breeding season (January), and immediately before the onset of the next breeding season (June) as determined from female reproductive status. Semen was collected by electro-ejaculation and analysed for volume, sperm number and motility characteristics. Ejaculate volume, total ejaculate sperm number, percentage motile sperm, and the sperm motility rating and index were all significantly elevated in September and significantly lower in November and January. This correlated with a significant increase in body weight, peri-cloacal gland width, and the weights of the prostate, Cowper's glands, urethral bulb and crus penis. The data confirm that male reproduction in the southern hairy-nosed wombat, like that of the female, is highly seasonal with a peak in reproductive capacity occurring in August–September and a reduction by November.

This study investigated burrow use and ranging behaviour in the southern hairy-nosed wombat Lasiorhinus latifrons in semi-arid South Australia. Sixteen adult wombats were fitted with radio transmitters and monitored monthly from July 2001 to February 2002. Wombats generally used between one and five warrens, preferred large warrens with a greater number of entrances and showed a preference for one or two warrens. Across the study period there was no apparent change in burrows used within warrens. Radio-tracking indicated that animals spent very little time above ground (26% of 1115 night-time fixes), centred their activity around their preferred warrens, and moved, on average, 99 m/h and 221 m/night. Mean home-range size, estimated using minimum convex polygons and the harmonic mean method from location data, obtained through triangulation, and daytime warren fixes, ranged from 1.3 to 4.8 ha. Home-range size was similar between males and females and home ranges overlapped substantially. The data highlight the importance of burrows to southern hairy-nosed wombats in shaping their home ranges. It seems likely that the use of burrows and a specialized diet are important energy saving strategies for this species in such unpredictable regions of South Australia.

A trapping study of five mammal species in wet sclerophyll forest adjacent to rain forest in the Australian Wet Tropics was used to examine the seasonal diversity, abundance and dung-specificity of dung beetles associated with mammal dung. A total of 542 dung beetles from 11 species within three genera was recovered from beneath the traps of 1104 mammal captures. The diversity of beetles associated with the dung of the northern bettong (Bettongia tropica), a mycophagous marsupial, differed significantly from the diversity predicted by a null model. Numbers of beetles varied significantly with type of dung, indicating preference by beetles. Beetle numbers were related positively to a 1-mo lag in monthly mean minimum temperature and less strongly to maximum temperature and rainfall. Significantly more beetles per mammal capture were detected in the wet season than in the dry season. Dung beetles showed a strong preference for either the Eucalyptus woodland (six species) or the adjacent Allocasuarina forest (four species), with only one species occurring in both habitat types. Beetle species from the Eucalyptus woodland were typically only detected in the late wet and early dry seasons, while species in the wetter Allocasuarina forest were generally collected during the late dry and early wet seasons. A significant ‘checkerboard’ species effect was detected in both time and space in both habitat types, suggesting that competition for dung was strong.

Sequence variation within 3 morphologically defined species of the anoplocephalid cestode genus Progamotaenia (P. ewersi, P. macropodis and P. zschokkei) was investigated using the cytochrome c oxidase subunit 1 gene. The magnitude of genetic variation detected within each morphospecies suggests that, in each instance, several cryptic species are present. Within P. ewersi, 5 genetically distict groups of cestodes were detected, 1 shared by Macropus robustus and M. parryi in Queensland, 1 in M. agilis from Queensland, 1 in Petrogale assimilis from Queensland, 1 in Macropus fuliginosus from South Australia and 1 in Wallabia bicolor from Victoria. In P. macropodis, cestodes from M. robustus from Queensland, Western Australia and the Northern Territory, M. parryi from Queensland and M. eugenii from South Australia were genetically distinct from those in Wallabia bicolor from Queensland and Victoria and from M. fuliginosus from South Australia. P. zschokkei consisted of a number of genetically distinct groups of cestodes, 1 in Lagorchestes conspicillatus and L. hirsutus from Queensland and the Northern Territory respectively, 1 in Petrogale herberti, P. assimilis and M. dorsalis from Queensland, 1 in Onychogalea fraenata from Queensland, 1 in M. agilis from Queensland and 1 in Thylogale stigmatica and T. thetis from Queensland. In general, genetic groups within each morphospecies were host specific and occurred predominantly in a particular macropodid host clade. Comparison of genetic relationships of cestodes with the phylogeny of their hosts revealed examples of colonization (P. zschokkei in M. agilis) and of host switching (P. zschokkei in M. dorsalis).