Potorous Gilbertii Research Articles

Introduction to the special issue of The Natural History of Two Peoples Bay Nature Reserve, Western Australia

Context This paper introduces the special issue of Pacific Conservation Biology devoted to the natural history of Two Peoples Bay Nature Reserve on the south coast of Western Australia. Methods This paper provides the background to the special issue. Key results Two Peoples Bay Nature Reserve was gazetted in 1967 for the conservation of the native biota, including two species; the Djimaalap/noisy scrub-bird (Atrichornis clamosus) and Ngilgaitch/Gilbert’s potoroo (Potorous gilbertii), both believed extinct for over 100 years before being rediscovered on the Reserve. The Reserve is 4774.7 ha in area, with wetlands, heathlands, granite outcrops, sand dunes, beaches, cliffs, and islands. Since it was established, mean annual rainfall has decreased by 16.8%, mean annual maximum temperature has increased by 0.2°C, and mean annual minimum temperature has increased by 0.7°C. Conclusions The paper poses the question: what do the changes of drier winters, hotter summers, and more extreme weather events mean for managers of conservation areas such as Two Peoples Bay? Implications Changing climate will pose problems for the managers of Two Peoples Bay Nature Reserve in ensuring the conservation of the Djimaalap/noisy scrub-bird and Ngilgaitch/Gilbert’s potoroo.

Sequence analyses at mitochondrial and nuclear loci reveal a novel Theileria sp. and aid in the phylogenetic resolution of piroplasms from Australian marsupials and ticks.

The order Piroplasmida encompasses two main families: Babesiidae and Theileriidae, containing tick-borne pathogens of veterinary and medical importance worldwide. While only three genera (Babesia, Cytauxzoon and Theileria) comprising piroplasm parasites are currently recognised, phylogenetic studies at the 18S rRNA (18S) gene suggest that these organisms represent at least ten lineages, one of which comprises the relatively unique and highly diverse Theileria spp. from Australian marsupials and ticks. As an alternative to analysing 18S sequences alone, sequencing of mitochondrial genes has proven to be useful for the elucidation of evolutionary relationships amongst some groups of piroplasms. This research aimed to characterise piroplasms from Australian native mammals and ticks using multiple genetic markers (18S, cytochrome c, oxidase subunit III (cox3) and cytochrome B (cytB)) and microscopy. For this, nearly complete piroplasm-18S sequences were obtained from 32 animals belonging to six marsupial species: eastern bettong (Bettongia gaimardi), eastern quoll (Dasyurus viverrinus), eastern grey kangaroo (Macropus giganteus), swamp wallaby (Wallabia bicolor), quokka (Setonix brachyurus) and Gilbert’s potoroo (Potorous gilbertii). The organisms detected represented eight novel Theileria genotypes, which formed five sub-clades within the main marsupial clade containing previously reported Australian marsupial and tick-derived Theileria spp. A selection of both novel and previously described Australian piroplasms at the 18S were also successfully characterised, for the first time, at the cox3 and cytB loci, and corroborated the position of Australian native theilerias in a separate, well-supported clade. Analyses of the cox3 and cytB genes also aided in the taxonomic resolution within the clade of Australian Piroplasmida. Importantly, microscopy and molecular analysis at multiple loci led to the discovery of a unique piroplasm species that clustered with the Australian marsupial theilerias, for which we propose the name Theileria lupei n. sp.

Balanoposthitis, Dyspareunia, and Treponema in the Critically Endangered Gilbert’s Potoroo (Potorous gilbertii)

The Gilbert's potoroo (Potorous gilbertii) is one of Australia's most critically endangered mammals with a current estimated population of 70 individuals. Both the wild and captive populations have a long history of balanoposthitis with associated crusting, ulceration, and preputial discharge. We sought to identify the microbial species found in the discharge, determine their significance in causing balanoposthitis, and correlate these findings with reproductive success and survivorship. Bacteriologic examination revealed the discharge to be a polymicrobial infection involving Treponema spp., Actinobacillus spp., and Pasteurella spp. Preputial histopathology reported a moderate, chronic, erosive inflammatory response with diffuse, moderate to marked secondary epithelial hyperplasia in conjunction with moderate numbers of spirochetes, suggesting a causative relationship. Clinical examination, preputial biopsies, and serologic screening found no evidence of associated systemic disease. The clinical investigation of Treponema is significant with respect to the overall recovery of Gilbert's potoroo, given the clinical and histopathologic similarities to Treponema paraluis-cuniculi found in rabbits, causing dyspareunia, and the severity of the associated balanoposthitis.

Monitoring reproduction in the critically endangered marsupial, Gilbert’s potoroo ( Potorous gilbertii): Preliminary analysis of faecal oestradiol-17β, cortisol and progestagens

Gilbert’s potoroo ( Potorous gilbertii) was rediscovered in 1994 after having been presumed extinct for 120 years. Estimates indicate fewer than 40 individuals remain at Two Peoples Bay Nature Reserve on the south coast of Western Australia although a translocated population of approximately 20 animals has recently been established on nearby Bald Island. A captive breeding facility has been established adjacent to the mainland population but few young have been produced (8 since 1995). Faecal levels of oestradiol-17β (E 2) were monitored over a 2-year period in an effort to determine cyclic reproductive activity, and faecal cortisol levels were also monitored to gauge whether chronic stress may be a factor limiting breeding in captivity. Faecal steroids were monitored in six captive females, and four captive male potoroos, and four wild females. The only captive births recorded after 1998 were one in August 1999 and one in February 2001, both to the same female. Peaks in E 2 concentration, up to 10 ng g −1 of dried faecal mass were measured and results to date suggest the main breeding period to be November–December based on elevated E 2 levels at this time. Clear patterns of reproductive activity in the captive females, however, were not evident. Analysis of epithelial cell counts from urinogenital swabs and faecal E 2 and progestagen (PM) levels from a single female kept at the Perth Zoo, suggest that Gilbert’s potoroo has an oestrous cycle of approximately 39 days. Faecal cortisol levels in captive females were significantly lower than those in wild-caught individuals and thus there is no indication that elevated cortisol levels per se inhibited reproduction in captive females.

Hematological and Serum Biochemical Reference Values and Cohort Analysis in the Gilbert's Potoroo (Potorous gilbertii)

Hematology and serum biochemistry blood values are tabulated for Australia's most critically endangered mammal, the Gilbert's potoroo (Potorous gilbertii). Significant differences were found between origin (captive or wild individuals) and age (subadult or adult) of animals. Sex, and presence or absence of Treponema infection, had minimal significance on blood values. Typical cell morphology is discussed, and hemoparasite examination identified Theileria spp. and Breinlia spp. Eighty samples were collected from a population of only 35 individuals, reflective of a population census rather than of a study reliant on statistical extrapolation. These reference ranges and findings will assist in the ongoing health management of this critically endangered species. hematology, biochemistry, marsupial, Gilbert's potoroo, Potorous gilbertii.

Morphological and molecular characterization ofTrypanosoma copemanin. sp. (Trypanosomatidae) isolated from Gilbert's potoroo (Potorous gilbertii) and quokka (Setonix brachyurus)

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.

Theileria gilberti n. sp. (Apicomplexa: Theileriidae) in the Gilbert's Potoroo (Potorous gilbertii)

The morphology and genetic characterisation of a new species of piroplasm identified in the blood of the Gilbert's potoroo (Potorous gilbertii) from the Two Peoples Bay Nature Reserve near Albany, Western Australia, is described from blood and tissue samples from 16 Gilbert's potoroos. Microscopy of blood showed these parasites are highly pleomorphic with a mean length of 1.8 mum and mean width of 0.85 mum. Phylogenetic analysis of 18S rRNA sequence data identified the piroplasm as a new species of Theileria that is closely related to other Australian marsupial piroplasm species. Based on biological and molecular data, it is proposed that the parasite from Gilbert's potoroo be given the name Theileria gilberti n. sp.

Fungi consumed by translocated Gilbert's potoroos (Potorous gilbertii) at two sites with contrasting vegetation, south coastal Western Australia

Gilbert’s potoroo (Potorous gilbertii) was previously widespread in south-west Australia but is now restricted to one location – a granitic area of shrubby heath at Two Peoples Bay Nature Reserve on the south coast of Western Australia. To alleviate the threat of extinction a program is under way to establish potoroos in other locations. At Two Peoples Bay Gilbert’s potoroos feed almost exclusively on truffle fungi. However, it was not known whether potoroos translocated to any new areas would be able to rapidly access and consume fungi after translocation, or whether truffles could be a sustainable food supporting breeding populations of potoroos in translocation sites. Furthermore, it was not known whether translocation of potoroos would be successful only in areas with vegetation similar to their refuge at Two Peoples Bay. The current study addressed these questions by analysing the diet of potoroos translocated into two contrasting areas – one on Bald Island with similar topography and vegetation to that of Two Peoples Bay, and the other a 14-ha enclosure on the mainland dominated by Eucalyptus–Allocasuarina woodland. The diet of potoroos was characterised by microscopic examination of scats from individuals trapped after their translocation to these areas. At Bald Island a diverse range of fungi was consumed immediately after translocation. Four potoroos released onto the island only 4–8 days previously consumed 23 species of fungi. Consumption was sustained over time. Three potoroos released onto Bald Island 1–2 years previously and one island-born individual consumed 27 species of fungi during a two-day sampling period. Nine of the 27 fungi species were the same as those that had been consumed by the potoroos within days after their release onto the island. This indicates that production of fungi and their consumption by potoroos on the island was sustained at least 1–2 years after translocation. Potoroos bred on Bald Island during this period. During the same period, two potoroos that were moved from captivity to the mainland site (Ryedene) were consuming six species of truffles within 29 days after release, and 14 species within three months. Such data indicate that a wider selection of vegetation types and areas than just those similar to where potoroos occur at Two Peoples Bay may be able to sustain potoroos and should be investigated for future translocations.

CRYPTOCOCCOSIS IN GILBERT'S AND LONG-NOSED POTOROO

Two cases of fatal cryptococcosis are described, one of Cryptococcus neoformans infection in a Gilbert's potoroo (Potorous gilbertii) and one of Cryptococcus gattii infection in a long-nosed potoroo (Potorous tridactylus). The diagnoses were confirmed by culture and specific immunohistochemistry, respectively. The long-nosed potoroo tested positive using the latex cryptococcal antigen test (LCAT), whereas the Gilbert's potoroo had a negative LCAT result despite having advanced disease of some duration. In both cases, the clinical presentation was a progressive neurologic disease associated with a central nervous system infection. Pulmonary infection was also observed in the long-nosed potoroo. Specific treatment with antifungal agents was unsuccessful in the long-nosed potoroo.

A quantitative dietary study of the ?Critically Endangered? Gilbert?s potoroo Potorous gilbertii.

Faecal analysis from the only known population of Gilbert?s potoroo (Potorous gilbertii) near Albany, Western Australia revealed that it, like other rat-kangaroo species is primarily mycophagous. Diet was determined by faecal collections from live-captured animals within Two Peoples Bay Nature Reserve. Microscopic examination of samples collected from June - September 2000 and additional samples from storage, found fungi to comprise over 90% of faecal matter. A total of 44 fungal spore types were identified with many believed to be of hypogeous origin. Fungal spores belonging to the genera Mesophellia, Elaphomyces, Hysterangium and an unknown spore type (Unknown 1) were frequently recorded in samples. Non-fungal material including plants (stems, roots and seeds) and invertebrates represented the remainder. This investigation found that P. gilbertii fed almost exclusively on fungi and could be considered a specialised mycophagist.

Chromosome morphology in Gilbert's potoroo, Potorous gilbertii (Marsupialia : Potoroidae)

Chromosome morphology was examined for male and female Gilbert’s potoroo, Potorous gilbertii, to infer taxonomic and evolutionary relationships among the extant taxa within the genus Potorous. P. gilbertii has the same number of chromosomes as P. tridactylus, 2n = 12,13. Giemsa-banding patterns were very similar in P. gilbertii and P. tridactylus; however, differences were noted between the sex chromosomes. Given that the relationships among extant Potorous are unresolved, we mapped karyotypes onto two alternative phylogenies to suggest methods of karyotype evolution within this group. Karyotypes and molecular-based information from the now ‘presumed extinct’ P. platyops or sequencing of multiple gene regions for phylogenetic analysis within the Potoroidae would provide valuable information for resolving the issue of rooting, and hence drawing conclusions on the evolution of karyotypes within this group.

Phylogenetic Relationships Within the Genus Potorous (Marsupialia: Potoroidae) Based on Allozyme Electrophoresis and Sequence Analysis of the Cytochrome b Gene

Controversy over the taxonomic designations within the genus Potorous has highlighted problems associated with using only morphological data. The recent rediscovery of Gilbert's Potoroo has allowed us to reexamine the relationship of this species to other extant potoroos. Phylogenetic relationships and estimated divergence times are presented, based on electrophoretic and cytochrome b sequence data. There was considerable concordance between the data sets. We conclude that Gilbert's Potoroo is a separate species and should be referred to by its original name, Potorous gilbertii (Gould 1841). Estimates for potoroid divergences are in line with those of Flannery (1989).